c\ 4897 b Texas Tech University Health Sciences Center SCHOOL OF MEDICINE / Department of Preventive Medicine and Community Health National Pesticide Telecommunication Network (NPTN) Lubbock, Texas 79430 (806) 743-3098 1-800-858-7378 S e p te m b e r 7 , 1988 Ben Nguyen C a r t w r i g h t , S l o b o d in , e t a l , C o u n s e lo r s a t Law 101 C a l i f o r n i a S t r e e t , S u i t e 2600 S an F r a n c i s c o , C a l i f o r n i a 94111 D e a r M r. N guyen: I am e n c l o s i n g c o p ie s o f l a b e l s f o r E s t e r o n 9 9 , F o rm u la 4 0 , a n d E s t e r o n 245. I am a l s o i n c l u d i n g i n f o r m a t i o n on 2 ,4 - D , 2 ,4 ,5 - T a n d S iv e x w h ic h a r e t h e c h e m ic a ls i n t h e s e . The c o n t e n t s o f e a c h h e r b i c i d e y o u l i s t e d i s b e lo w : E s te ro n K uron F o rm u la 2,4-D ow E s te ro n 245 - 2 ,4 ,5 - T S i 1v ex 40 - 2 ,4 -D Weed K i l l e r ( I c o u ld n o t f i n d t h i s p r o d u c t ) 99 - 2 ,4 -D We do n o t h a v e some o f t h e i n f o r m a t i o n you r e q u e s t e d . I t h i n k Freedom o f I n f o r m a t i o n w ould be y o u r b e s t s o u r c e o r t h e R e g i s t r a t i o n D i v i s i o n o f t h e E n v ir o n m e n ta l P r o t e c t i o n A g en cy . The a d d r e s s e s a r e b e lo w . Freedom o f I n f o r m a t io n F 0 I O f f i c e r (A -101) USEPA 401 M S t r e e t , SW W a sh in g to n , DC 20460 R ic h a r d M o u n tf o r t R e g i s t r a t i o n D i v i s i o n (T S-767C ) USEPA 401 M S t r e e t , SW W a s h in g to n , DC 20460 T h an k you f o r w r i t i n g a n d l e t me know i f P e s tic id e S p e c ia lis t I c a n be o f a n y f u r t h e r h e l p . V The LDso and How To Interpret It Pesticide toxicologists use rather simple' anim al toxicity tests to ran k pesticides according to their toxicity. L o n g before pesticides are registered w ith th e Environm ental Protection Agency and eventually released fo r public use, th e m anufacturer m ust delcare th e toxicity o f their pesticide to the w hite ra t under laboratory conditions. T his toxicity is defined by the LD3o expressed as milligrams (mg) o f toxicant per kilogram (kg) o f body weight, the dose th at kills SO percent o f the test anim als to which it is adm inistered under experim ental conditions. T he LDso is m easured in term s o f oral (fed to , o r placed directly in th e stom achs o f rats), derm al (applied to the skin o f rats o r rabbits), and respiratory toxicity (inhaled). Combinad tabulation of pesticide toxicity classes. Routes of absorption ld m Toxicity rating Single oral dose for rats, mg/kg LDM Single dermal dose for rabbits mg/kg Probable lethal oral dose for man ' 6—Supertoxic 5— Extremely toxic A—Very toxic 3—Moderately toxic 2—Slightly toxic 1—Practically nontoxic <5 5« 50-500 500-5,000 5,000-15,000 > 15£00 <20 20-200 200-1,000 1,000-2,000 2^00-20,000 >20,000 A taste, a grain A pinch, 1 teaspoon 1 teaspoon to 2 tablespoons 1 ounce to 1 pint 1 pint to 1 quart >rt quart Source: Toxicity ratings modified from M. N. G leason, R. E . G leason, and H. C . Hodge. 1976. ¿Sinica/ Toxicology o f Commercial Products. 4th ed. W illiam s and W ilkins Company, Baltimore, M d.p.6 * In LDW tests, 100% concentrates of the chem ical are used (unless otherwise stated), w hereas when you use the chem ical, it is diluted to a lower concentration which has a lower toxicity. Exam ple: Carbary! L D * m 500 mg/kg - Moderately to Very toxic 5% Savin Oust-*- LOM - 10,000 mg/kg - Slightly toxic (Carbary! is active Ingredient) The P esticide Book George W . W are. W. H. Freeman & Co. San Fran cisco - 1976 ( 4899 • i FOR THE CONTROL OF TREES, BRI : .1 Low-Volatile Brush and W eed Herbicide for Industrial ACTIVE INGREDIENT: 2,4,5-Trichlorophenoxyacetic Acid, Propylene - Glycol Butyl Ether E ste rs........................................ ..............................69.2% INERT INGREDIENTS: .......................................... . . . .......... ....................30.8% 2,4,5-Trichlorophenoxyacetic Acid Equivalent —45-0% . >M 4 Pounds per Gallon . E.RA. Registration No. 464-205 l - ; E.P.A. E s t 464-MI-1 ; t ;i - JS - 3 PRECAUCION AL,-USUARIO: Si usted no lee inglés, no use este producto hasta que laetiquéta le haya sido explicada ampliamente. TRANSLATION:'(TO THE USER: If you cannot read English, do not use this product untiltheíábel has been fully explained to you.) ; t. "t-J •# - i - ■• ñ t V . h t i *.< w 1^ ' i f 1 1 lílífl; ¿ i ■\ i i T- V * .* - Í *■ JR * •; r V7 i-, j •* i f * £-'4^' > " • i* r - -.■■■ iS - X .■ ■i r l - ■■a i t i l l i i i i ' ; ; ■ i :■ i , ! r i ». - i " II 86-1064 PRINTED IN U . S . A . IfO E C E M B E R , 1 9 80 .. REPLACES SPECIMEN LABEL 86-1 Ob^TPRINTED O&TPRINTED It l JANUARY, 0/ 1980. DISCARD PREVIOUS SPECIMEN LA B ELS. REVISION S INCLUDE: REPRINTED TO CORRECT ERROR IN PRECAUTIONS; CONTAINER GRAPHICS WERE CORRECT; SPECIMEN LABEL OF JANUARY 1930 4900 P S C '$ i * o L A B E L for Industrial Vegetation Control, Fencerows, and Rangeland K E E P OUT O F TH E REACH O F CHILDREN ...69.2% ' MAY B E HARM FUL IF SW ALLOW ED • MAY C A U SE IRRITATION Avoid Contact with E y e s, Skin and Clothing / Do Not Cut or Weld Container , i \ In case of an emergency endangering life or I property involving this product, call collect | 517-636-4400 • A * I AGRICULTURAL CHEMICAL Do Not Ship or Store with Food. Feeds. Drugs or Clothing • , > , 5: iT • r * ■' •* - -}:i - i i l - : l ; 1 | Ü ' f ii '? x*1• •• '• :v r ; ? v 'w : ‘.1 -h f.&-a .*i■ }i if M : ) 5 a ■l i i .r-’ . ¿ a •• - • V ’ .« •r V‘ ■? k i ; -i i »■* : i -S >. i i. i } » t ; > f-; 4901 P - H llk FiON 245 HERBICIDE 1er Esters of 2.4,5-T • Acid Equivalent: 4 Pounds per Gallon gallons of ESTERON 245 in 100 gallons of oil. Brush of average i feet high may take up to ISO gallons of spray mixture per acre, nt: Where growth is more than 6 to 8 feet tall, cut it close to the i the freshly cut stumps and stubs with 3 gallons of ESTERON (1 pint in 4 gallons) of oil, mixed thoroughly. For more resistant tons of ESTERON 24S in 100 gallons II pint in 3 gallons! of oil. II exposed bark, as well as cut surfaces. This means spraying unn-off to the ground line is noticeable. Old or rough bark requires ne than young or smooth bark. Apply at any time, including ccept when ice. snow or water prevent spraying to the ground ire obtained on freshly cut stumps two inches across or larger, je normally requires from 10 to 100 gallons per acre depending nps and stubs. it: For large trees, make a singlehack girdle or "frill" of crvercompletely around the tree as close to the ground as feasible, »roughly using a mixture of 2 gallons of ESTERON 245 in 100 n 3 gallons) of oil. latm ent: Use 1/4 pint of ESTERON 245 in 3 gallons of water all foliage, shoots, stems and bark without runoff. LO W V O LU M E S P R A Y S tc sprays containing ESTERON 245 when foliage is well nts are actively growing. For best results on woody species, soil e sufficient to promote foliar growth. Spraying during proionger or after leaves have lost their normal green color and vigor sfactory control. Apply low volume sprays by air or ground hen spray drift will not be a problem —note use precautions. Usi w ered Knapsack Sprayer—Mix 1 1/2 to 2 gallons with ^ \ or kerosene to make 20 gallons of total spray soluportab. _ .tapsack mistblower to all sides of lower brush stems collar. Good coverage of the root collar is essential for best Slower at 1/4 to 1/3 throttle for best spray delivery and ximum drift control use a basal nozzle attachment and do not the horizontal position. I A P P LIC A T IO N FO R B R U S H C O N T R O L »Itural Experiment Station, your local Extension Service Weed ts for best time to treat and need for re-treatment in your area, arty boot to milk stage where grass seed production is desired. >int ESTERON 245 plus 1/2 to 1 gallon of oil in enough water to I total spray per acre. Apply 40 to 90 days after first leaves ak: Use 1/2 to 1 quart of ESTERON 245 plus 1 gallon of oil in nake 4 gallons of total spray per acre, ck O aks: Use 2 quarts of ESTERON 245 plus 1 gallon of oil in nake 4 to 6 gallons of total spray per acre. U S E PR ECA U TIO N S e dairy animals on treated areas within 6 weeks after applicameat animals on treated areas within 2 weeks of slaughter, rr WITH 2.4.5-T SUSCEPTIBLE CROPS AND OTHER >AOLEAF PLANTS—ESTERON 245 Herbicide is injurious to mts. Therefore, do not apply directly to or otherwise permit nts to contact cotton, grapes, tobacco, fruit trees, vegetables. »Is or other desirable plants susceptible to 2.4.5-T. Do not use ■ ■. V 3 ’" N « \ CONTAINS BUTOXYETHYI For the Control of Many Broadleaf Wee and Woody Plants Susceptible to 2,4-D in Grass Past ACTIVE INGREDIENT 2.4- Dlchlorophenoxyacetic add, Butoxyethyl E s t e r f ......................................................62.5% INERT INGREDIENTS ..........................................................37.5% 2.4- Dichlorophenoxyacetic Acid Equivalent: 43.2% -3.8 lb/gal tlsom er Specific by AOAC Method No. 6275-6.279 (13th Ed.) EPA Reg. No. 464-566 EPA Est.464-MI-1“ *; 359-OR-15' Superscript used corresponds to letters in LOT number. Avoid Contac In case of contact. irritation persists. H sticking finger down t unconscious person. PRECAUCION A L U SUARIO: S i usted no lee inglés; no use este producto hasta que la etiqueta le haya sido explicada ampliamente. TRANSLATION: (TO TH E U S E R : If you cannot read English, do not use this product until the label has been fully explained to you.) • „ j 1 8 . 9 3 L / 5 This product istoxii likelv to occur. Do g a l Î it ' * f ’# : .• 1 ? : I •$ í 86-1674 PRINTED IN U .S .A . IN; SEPTEMBER, 1982". REPLACES SPECIMEN LABEL 86-1674~PRINTED IN MAY, 1982. DISCARD PREVIOUS SPECIMEN LABELS. REVISIONS INCLUDE: (1 ) REVISED WEED L IS T , "USE IN LIQUID NITROGEN F E R T IL IZ E " SECTION, "WEED C 0 T P 0 L IN SMALL G R A IN S ..." E r CTICN, USE PREC-J~I0N.., AND STORAGE ANO DISPOSAL INSTRUCTIONS ' _ ) ADDED 4904 H _cjeCt 4 1 oid Contact w ith Skin, Eyes, or Clothing • W ash Thoroughly After Handling j Statements of Practical Treatment ise of contact, iipmediately flush eyes or skin with plenty of water. Get medical attention if ition persists. If swallowed, induce vomiting immediately by giving two glasses of water and ing finger down throat. Call a physician. Do not induce vomiting or give anything by mouth to an insrious person, j Physical o r Chem ical Hazards ■ , ' j Do Not Cut or Weld Container ■ ; i ; •1 Environm ental Hazards product is toxic tp fish. Do not apply directly to any body of water. Do not apply where runoff is i to occur. Do nqt contaminate water by cleaning of equipment or disposal of wastes. Do not aminate irrigation ditches or water used for irrigation or domestic purposes. <■' case of an emergency endangering life or opetty involving khis product call collect: ¿■517*636-4400 > •; rvjr 5 : : ■■ : jg h or denting stag«, apply 1 to 2 pints erenm ai w eeds, decrease weed seed •d. cockfebur. dogbane, jimsonweed. with ‘ 'westing Do not torage or feed 'U S t JR A IN F IE L D S : Following the num. w ild g arlic and wild onion often 2 to 3 quarts per acre of ESTERO N 99 3«rt of a control program . Do not torage -*r acre when sorghum is S to 15 inches ' to control som e w eeds but the chance it. Do not treat before the sorghum is S jg h stages. If sorghum is taller than 6 « a s m uch a s possible. Temporary crop re and high a ir tem peratures Varieties -isitrwe Spray only varieties known to be .Mural Experim ent Station or Extension 9 am ount of w ater required for unrform iished stands in spnng from the tiller to seeding« may be treated with the lower ¿1 w eed regrow th may be treated m the .M ASS P A STU R ES: N O TE: Do not graze ation. D o not use on bent grass, allalta. areas until g rass « w e ll established. Do 3 production is desired. i-helder. MusktMstfte and Other B ra e * oer acre in the amount of water needed growing actively. 1 quart per acre win *1 w eeds may require repeated treatm aking three applications (tau-spnngrm g. »grass: Apply t to 2 quarts per acre use 1 quart m 5 gallons of oit or m 4 ircraft between May 15 and June is . On ->dapply by aircraft when foliage is fully acre m 2 to 3 gallons of oil or in 3 to 5 e 3 Quart rate is usually required. Brush d Retreatm ent may be needed, tebruah and C ertain Other Chaparral 3f w ta r. One gallon of fuel oil may be Jake applications by aircraft or ground ctive control, the brush m ust be fully nent m ay be needed, co p e o es susceptible to 2.4-0 m *\ anks. spray brush up to 5 to ft 1 4 Qw_. is of ESTER O N 99 Concentrate including foliage, stem s and berk. Ttua auate co verage of solid stand of brush. m e sp ray off the area being treated, ore frost as long as the so il moisture a esa effective m midsummer during hot ire net atfrvety growing. O il or wetting sd effectivene ss. ad leaf w eeds and brush on sites to be STERO N 99 Concentrate herbicide in ty 6 to 25 gallons. Applications can be apsack sprayer). Two to eight quarts of -i«ybe added to improve brush control, -iter o r spring to contro l susceptible rs. c u r a r e , cherry, service cherry and ;r«ak. u se ESTER O N 99 C oncentrate« >r ground in sufficient spray volume tor in plantatio ns w here pete or tench era )hbte evergreen brush sp ecies, euch as m anzanita o r deciduous brush after its at rates up toftquarts per aere alone a o il or suggested rates of suitable STERO N 9ft Concentrate without o«L -tart» per acre but may cause injury or •» of sp ray tor good coverage of brush, •enouaiy injured by treatment at these n«. lack pm e. red pine, black spruce. * a harden oft and brush »stiH actively * Concentrate per acre m enough water * ground to control certain competing 0nd w ihow Sm ce ttus treatment may >i cannot be tolerated. " • ): Apply ESTER O N 99 Concentrate * <*ptg grasses such as b»nt and ■** nin g rass is waff esrat htned m l With spring application *>g v - ju m es ara usually damaged or • riaeor *ot»»d perenmai w eeds may u ~qt on- r a r e . E 4 T E R C N - 9 Concentrate herbicide - e * ? l O ca' m s of water Spray to wet all foliage (400-800 gallons per acre). Addition of a wetting agent may be advisable Apply m the spring during flower head em ergence. Respray it needed when regrowth is 3 to 5 feel tall W EED CON TRO L ON FALLOW LAND: Use 1W to 2 % quarts per acre on annual broadleaf weeds and up to 3 quarts per acre on established perennial species, such as Canada thistle and lie u bindweed. Apply to actively growing weeds. Do not plant treated tallow land until three months alter treatm ent, or until chem ical has disappeared from soil. SPO T TREATM EN T: To control broadleaf weeds in small non-cropland araas with a hand sprayer, use Vi pint of ESTERON 99 Concentrate in 3 gallons of wstar and spray to thoroughly wet alt weed foilage. Keep spray m ixture agitated to prevent separation. CO N TRO L O F WOOOY W EEDS IN LO W -BU SH BLU EBERRY FIELD S IN M AIN E: How to u se: Mount a drum ft to 10 feet long or som e other suitable length, and 1Vr to 2 feet in diam eter on an axle su ch as an ok} hay raka frame. Cover the drum with watar absorbent yet tough cloth which win resist rapid wear and tear. Draw the cloth-covered drum across the blueberry field and at the sam e time spray evenly onto the full length of the top of the cloth-covered drum a spray m ixture made by diluting 1 quart of ESTERO N 99 Concentrate in 50 gallons of water per acre. Have the drum mounted so that aa it revolves on its axis it is high enough to m iss most of the low bush blueberry stem s, yet low enough to forcibly brush the spray-saturated cloth-covered drum against the higher woody w eeds, principally sweet fern, wild cherry and poplar. Kaep the cloth wet enough to provide top coverage of the weeds, yet not so wet as to allow runoff of the liquid w hich could cause m/ury to the blueberry plants. W hentoU se: Apply during June and Ju ly when weed tops have amarged sufficiently above the blueberry stem s to allow treatment ot the weeds and not the blueberry plants. Apply only during the year before the first bum. To use this method of weed control, two-year bum s should be extendad to three years. Caution: Do not allow tha spray baing applied to the cloth-covered drum to be directed onto the blueberries. Do not harvest-rake field during the herbicide treetment year or until a two-year interval thereafter. *0 *0 ?5<% U SE PRECAUTIONS AVOID CON TACT WITH 2,4-0 S U SC EP TIB LE CRO PS AND OTHER D ESIR A B LE BRO AD LEAF PLA N TS: Do not apply directly to or otherwise permit even minute am ounts to contact cotton, grapes, tobacco, fruit trees, vegetables, ftowers, ornamentals or other desirable plants susceptible to 2.4-D. Do not use in or near a greenhouse. DO NOT APPLY IN THE VICINITY O F COTTON. GRAPES. TOBACCO. TOMATOES OR OTHER D ESIR A B LE 2.4-D SU SC EP TIB LE CRO PS OR PLANTS. DO NOT SPRAY WHEN WINO IS 8LOW ING TOWARDS SU SC EP TIB LE CRO PS OR ORNAMENTAL PLANTS. AVOID SPRAY D R IFT: Applications should be made only when there is no hazard from spray drift since very sm all quantities ot spray, which may not be visible, may severely injure susceptible crap s during both growing and dormant periods. Use coarse sp rays to minim ize drift sin ce, under adverse weether conditions, fine spray droplets may drift a m ile or more. A spray thickening agent sweh as NALCO TRO L1. may be used with this product to aid m reducing spray drift. If used, follow all use recom m endations and precautions on the product lab el ’ NALCO TR O L — Trademark of NALCO Chem ical Company GROUND EQUIPM EN T: With ground equipm ent spray drift can be lessened by keeping the spray boom a s low as possible: by applying 20 gallons or more of spray per acre: by using no more than 20 pounds spraying pressure at large droplet producing nozzle tip s: by spraying when wind velocity is low: and by stopping all spraying whan wind exceeds 6 to 7 m iles per hour. Do not apply with ho&ow cone-type insecticide or other nozzles that produce a bnedroplet spray DETERM IN E AtR MOVEMENT AND D IRECTIO N S B EFO RE FO U A R APPLICA TIO N : U se a sm oke generator or other means at or near the application site for the detection of air movement a * stability or temperature inversions. Such a condition exists when there is Httte or no wind and air temperature n lower near the ground than at higher levels. U se appropriate drift control m easures or avoid application when sm oke a moving toward nearby desirable susceptible plants or sensitive areas. A ER IA L APPLICA TIO N : With aircraft, drift can be les sened by applying a coarse spray; by using no more than 20 pounds spray pressure at tha nozzles: by using straight stream nozzle» directed stream back: by using e spray boom no longer that Vi tha wing or rotor span of tha aircraft: and by spraying only when wind velocity is lew than ft mph. Exce ssive am ounts of this herbicide in the so il may temporarily inhibit seed germ ination or plant growth. Violent wind storm s may move sod particles. If 2.4-0 is on so il particles and they ara blown onto the susceptible plants, visible symptoms may appear. Serious injury is unlikely The hazard of movement of 2.4-0 on dust is reduced rf treated fields era irrigated d r rt ram o ccurs shortly after application. AT HIGH TEM PERA TU RES. VAPORS FROM TH IS PROOUCT MAY IN JU RE SU S C EP T IB LE PLAN TS GROW ING NEARBY. To avoid injury to desirable plants, do not handle or apply other agricultural cham icala with the same equipment used lor ESTERO N 99 Concentrate unlew appropriately cleaned fu st Le e « conditions may affect the use of herbicides. Consult your Stats Agricultural Experiment Station or Extension Service weed sp ecialists for cleaning methods which ara in com pliance with lo cal regulations and for advice in selecting treatments from this lab « to best fit local conditions. Be sure that use of this product conform s to «1 applicable regulation«. Apply this product only as specified on this tab«. )■ STORAGE AND DISPOSAL Do not contam inate water, food or feed by storage or disposal STO RA G E: Keep container tightly dosed when net in use. P ESTIC ID E D ISPO SA L: Pesticide, spray mixture or rinsafe that cannot be used according to label instructions must be disposed ot according to Federal. S t« e . or lo cal procedures w ider the Resource Conservation and Recovery A ct CON TAIN ER D ISPO SA L: Triple rinee or equlvalant and offer for recycling or reconditioning, or dispose o< in a sanitary landtift. or by other approved State and local procedures. M ONEY BACK GUARANTEE fT T fftfT N ft rnnrew trats heittirkte Is QuerantaerthyTtu Oow Chem ical Com pany to tne tun extent of the purchase p rice: 1. To give satisfactory central of n ee d and brash sp ecies listed on conta in er when 2. To form a aultabto spray m ixture In any water At for spray usn. 3. To store satisfactorily at tem peratures a s few as -4 (rF. KOTiCC Shu t «orrenfeihet tie product conforms lo m w w snd ttreaeonablyM ter Pie purpows m lM or' the laoei u m need ki eecorexrt* » thow tw i» .inoer normal condition* otuae. Out rvertT-vr -n» mirmntf nor any Other w m n>| of MEACHANTA8H T> or FITNESS FOR A PARTICULAR PURPOSE, -xpree» or xnohed. extend« lo ttw m of mu product corverr to laoel instruction*. or undsr epiofim l conditions, or under conditions not rawoneOh t >sailer and buyer assume« me nea o' any s>*ch wee 10596-025-2 20682-6992 ) THE DOW CHEMICAL COMPANY ANO SUBSIDIARIES M ID L A N D . M IC H IG A N 4B B 4 Q . U S A H O R G E N . S W U Z l R lA N O HO NGKO NG C O R A L G A B L E S . F LO R ID A 3 3 1 3 4 . U S A S A R N 'A O N T A R IO . C A N A D A 4906 * tradem ark ®f THE OOW CMC M ica i » OMPANr V - Î20O C > t * WEED LIST ESTERO N 99 Concentrate herbictoe t Treatm ent: Apply 1 to 2 oints iw< acrt when gram s a rt In the hard dough stag*' tc control large w eeds that may in tsrere w in n rv e sL Bast resude vrtH ba obtained when soim oisture is sufficient to cau sa suci uU*nt w«-ed growth. N O TE: Do not feed treated t v s * to livestock. W EED CON TRO L W CO RN : Use on»oMh»fc Mowing in ree program s. P tsem srg eo ce: Apr iy t to 2 quarts per acre to so il anynm»* •*!•> p *n:>ng but before com em erges Only em eraec twoedleafed w eeds are likely to be c o n to'ter Do not apply m ere than 1 quart per acre urnes* the increased risk ot crop injury can be to-e» iteo Do not use on light sandy soil. Em ergence: Apply 1 pmt per sere just as corn plants are breaking ground. Fetto m eig an ca: Alter emergence of com use to omt per sere Aop eatmn of to to 1 pmt per acre may be needed fo> m u inu m controlo! some weeds but such ra--s are mere «kieyto injure th sco rn .it com -sever 6 >c,'estali.ueedropp-5zrtosto-e*ot*,e>or sy ch fie oi j s - w :h *1 atiszm e or other adjuvant* C*oc m urv is more likely to occur il con- is gn w -c -a. >di. under high tem peraure and high so» moisture conditions. To reduce breakage >f*'» - s ■•on tpm voisryonttleness caused by 2.--D d- ia\ cultivation for 8 to 10 days afi-»r t*e.-.trr - - r 5 n:>t ‘orngeor feed com fodder tor 7 cay* fc io-vmg application, n o t e H>or*is . a w '• r m se t? r 4 -0 and som e are easily mju ec S : ra. only varieties known to be to e ra n to . - D ;©i tar t seed company or you* Agricultura £ •> 'im ent Station or Extension S**rv ce . sp-^ci ns-s *or this information V PREH A RV EST CORN TREATM EN T: After the hard dough or denting so per acre by ad wttiow. Sm ce cause occasio n al conifer mjury. do not use > ------------------A product of identical name containing pentachlorophenoi (Chapter 4) as } th e active ingredient has been discontinued by Amchem Products Co. '$ A product o f identical name marketed by the Crystal Chemical Com pany ^ contains cacodylic acid as the active ingredient (Chapter 10). § TOXICOLOGY Some o f the chlorophenoxy acids, salts, and esters are moderately irritating to skin, eyes, and respiratory and gastrointestinal linings. In a few individuals, local depigmentation has apparently resulted from prolonged ahd repeated dermal contact with chlorophenoxy materials. The chlorophenoxy compounds are absorbed across the gut wall, lung, and skin. They are not significantly fat storable. Excretion occurs within hours, or at most, days, primarily-in the urine. Given in large doses to experimental animals, 2,4-D causes vomiting, diar­ rhea, anorexia, weight loss, ulcers o f the mouth and pharynx, and toxic injury to the liver, kidneys, and central nervous system. Myotonia (stiffness and in­ coordination o f hind extremities) develops in some species and is apparently due to CNS damage: demyelination has been observed in the dorsal columns of the cord, and EEG changes have indicated functional disturbances in the brains o f heavily dosed experimental animals. Ingestion o f large amounts o f chlorophenoxy acids has resulted in severe metabolic acidosis in hum ans. Such cases have been associated with electro­ cardiographic changes, m yotonia, muscle weakness, myoglobinuria,' and ele­ vated serum creatine phosphokinase, all reflecting injury to striated muscle. Because chlorophenoxy acids are weak uncouplers of oxidative phosphoryla­ tion, extraordinary doses may produce hyperthermia from increased produc­ tion o f body heat. Polychlorinated DibenzoDioxin (CDD) compounds are generated in the synthesis o f 2,4,5-T. The 2,3,7,8-Tetra CDD form is extraordinarily toxic to multiple mammalian tissues. Hexa- hepta-, and octa-compounds exhibit less systemic toxicity, but are the likely cause o f chloracne (a chronic, disfiguring skin condition) seen in workers engaged in the manufacture o f 2,4,5-T, and certain other chlorinated organic compounds. Although toxic effects, notably chloracne, have been observed in m anufacturing plant workers, they have not been observed in form ulators or applicators regularly exposed to 2,4,5-T. The medical literature contains several reports o f peripheral heuropathy following what seemed to be m inor dermal exposures to 2,4-D. It is not certain that exposures to other neurotoxicants were entirely excluded in these cases. Single doses of 5 m g/kg body weight o f 2,4-D and 2,4,5-T have been administered to human subjects without any adverse effects. One subject consumed 500 mg of 2,4-D per day for 3 weeks without experiencing symptoms or signs o f illness. W ^ « O F r p-2j4;3-TPTSfl'Vtx)’is the propionic acid homologuc of 2,4,5-T.tKllrOh1S(«.low volatile ester o f 2,4,5-TP. 2,4,5-TB is the butyric acid homologue of 2,4,5-T. Fenac or chlorfenac is 2,3,6-trichlorophenylacetic acid. Dicamba (feanvel) is dichloroanisic acid. M CPA, M CPB, MCPB-Ethyl, MCPCA and M CPP (Mecoprop) are 2-methyl, 4-chlorophenoxy aliphatic acids and eslets. , p | ^ * (\j qq (\j h 0 0 ( < F R E Q ir - SYM P TO M S A N D SIG N S O F P O ISO N IN G Chlorophenoxy compounds are moderately IRRITATING to skin and mucous membranes. Inhalation o f sprays may cause burning sensations in the nasopharynx and chest, and coughing may result. Prolonged inhalation some­ times causes dizziness. When INGESTED, high concentrations of chlorophenoxy compounds may irritate the ipouth, throat, and gastrointestinal tract. Prom pt EMESIS, CHEST PAIN (from esophagitis), ABDOMINAL PA IN , and DIARRHEA commonly ensue. Injury to the GI tract does not usually progress to ulceration or perforation. Absorbed chlorophenoxy com pounds have caused FIBRILLARY MUSCLE TW ITCHING, skeletal muscle tenderness, and MYOTONIA (stiffness o f muscles of the extremities). Ingestion o f very large amounts has produced METABOLIC ACIDOSIS, fever, tachycardia, hyper­ ventilation, vasodilatation and sweating. Particular cases have been charac­ terized by coma and convulsions. CONFIRM ATION OF DIAGNOSIS Gas-liquid chromatographic methods are available for detecting and mea­ suring the chlorophenoxy compounds in blood and urine. These analyses are useful in confirming and assessing the magnitude o f chlorophenoxy absorp­ tion. Urine samples should be collected as soon as possible 'after exposure because the herbicides may be almost completely excreted in 24-72 hours, depending on the extent o f toxicant absorption. Analyses can be performed at special laboratories operated by state health departm ents, chemical com­ panies, universities, and government facilities. If circumstances indicate strongly that excessive exposure to any of these compounds has occurred, ini­ tiate appropriate treatment measures immediately, not waiting for chemical confirmation o f toxicant absorption. TREATMENT CO O 1. to 2. 3. \ o o 4. ■c: BATHE aqd SHAM POO with soap and water to remove chemicals from skin and hair. Individuals with chronic skin disease or known sensitivity to chemicals should either avoid using these herbicides or take extraordi­ nary measqres to avoid contact. FLUSH contaminating chemicals from eyes with copious amounts of clean water for 10-1S minutes. If symptoms o f illness occur during or following inhalation o f spray, REMOVE victim FROM CONTACT with the material for at least two days. Allpw subsequent contact with chlorophenoxy compounds only if effective respiratory protection is practiced. IF substantial amounts o f chlorophenoxy compounds have been IN­ GESTED, spontaneous emesis usually occurs. Ordinarily, this empties the stomach as effectively as intubation and lavage. If vigorous emesis 3. has not occurred and IF VICTIM IS FULLY ALERT, inouce EMESIS with SYRUP O F IPEC A C (adults 12 years and older, 30 ml; children under 12 years, 13 ml), followed by 1-2 glasses of water. Following emesis, administer 30-30 gm o f ACTIVATED CHARCOAL in a slurry of 6-8 ounces tap water, to limit absorption o f herbicide remaining in the gut. IF CONSCIOUSNESS LEVEL IS DEPRESSED or. other signs or NEUROTOXICITY appear, SUSPECT additional of alternative ingested toxicants. Evacuate the stomach by INTUBATION, ASPIRATIO N , and LAVAGE. Because petroleum distillates are commonly in­ cluded in chlorophenoxy form ulations, gastric intubation incurs a risk o f hydrocarbon pneumonitis from aspiration. For this reason: A. B. C. 6. 7. § (¡) ^ qj ^ 0 (\] If victim is unconscious o r obtunded and facilities are at hand, in­ sert an EN DOTRACHEAL TUBE (cuffed, if available) prior to gastric intubation. Keep victim’s H EAD BELOW LEVEL OF T H E STOMACH dur­ ing intubation and lavage (Trendelenburg, or left lateral decubitus, with head o f table lipped downward). Keep victim's head turned to left. ASPIRATE PHARYNX as regularly as possible to remove gagged or vomited stom ach contents. P I ^ * 01 83 After aspiration o f gastric contents and washing of stomach, instill 30-30 gm o f ACTIVATED CH A RCO A L in 3-4 ounces o f water through the stomach tube to limit absorption o f remaining toxicant. Do NOT instill milk, cream, or other materials containing vegetable or animal fats, as these are likely to enhance absorption. If bowel movement has not occurred in 4 hours and patient is fully con­ scious, give SODIUM SU LFA TE, 0.23 gm /kg, as a cathartic. Magne­ sium sulfate and citrate, in comparable dosages, are equally suitable if renal function is adequate. Retained magnesium may depress CNS func­ tion. In SEVERE POISONINGS by very large amounts o f ingested chlorophenoxy acids, forced ALKALINE DIURESIS may save the victim's life. Assess serum electrolyte concentrations, and serum and urine pH . If a metabolic acidosis is present, infuse solutions o f sodium bicarbonate at rates sufficient to keep the urine distinctly alkaline, continuing until plasma concentrations o f chlorophenoxy compounds are less than about 10 ftg/m l. (Prescott, L F. et al., Br. J . Clin. Pharmacol. 7:11 (1979)) T OO PESTICIDE & TOXIC m uu is - ÎS if ïiî' rf£2rti ° n h?z.ardous wastes, pesticides, toxic subi stances .and .general issues of. ^regulation and legislation W P a g e 10 O c to b e r 1 9 , 1983 PESTIC ID E & T O X IC CHEM ICAL NEWS E PA N O T E S IN T E N T T O E N D A L L U SES O F 2 ,4 ,5 - T A N D SILV EX F o llo w in g D ow C h e m ic a l C o m p a n y 's w ith d ra w a l fro m th e 2 ,4 ,5 - T a n d silv ex c a n c e lla tio n h e a rin g a n d v o lu n ta ry c a n c e lla tio n o f r e g is tr a tio n o f th e p r o d u c ts , EPA h a s issu e d n o tic e s t o o th e r r e g i s tr a n ts se e k in g to e n d a ll u s e s o f th e p e s tic id e s w ith o u t m o re h e a rin g s (S e e A u g . 1 0 , P a g e 5; a n d A ug. 3 1 , P a g e 2 ). T o c o n tin u e r e g i s tr a ti o n s , h e a rin g s h av e to be r e q u e s te d w ith in 30 d a y s o f th e a g e n c y 's O c t. 14 n o tic e . T h e O c t. 14 in te n t t o c a n c e l n o tic e c o v e rs a ll re m a in in g n o n -s u s p e n d e d r e g is tr a tio n s o f th e p r o d u c ts , th o s e la b e le d fo r r ic e , r a n g e , s u g a r c a n e , o r c h a r d an d m is c e lla n e o u s n o n c ro p u s e s , m a n u f a c tu r in g u s e , an d te c h n ic a l r e g i s tr a ti o n s . A t th e s a m e t im e , EPA issu ed a n e n f o r c e m e n t p o lic y s t a t e m e n t " t o p ro h ib it th e s a le , d is tr ib u tio n , im p o r ta tio n , o r o th e r t r a n s f e r o f u n r e g is te r e d 2 ,4 ,5 - T a n d silv ex p r o d u c ts , e x c e p t a s s p e c if ic a lly a llo w e d by F IF R A S e c tio n 3 ( b ) ." T he a g e n c y s a id it w as a ls o " re q u irin g e a c h r e g i s tr a n t o f a 2 ,4 ,5 - T o r silvex p r o d u c t to a m e n d th e c o n f id e n tia l s t a t e m e n t o f fo rm u la fo r t h a t p r o d u c t t o id e n tif y th e s o u rc e o f e a c h a c tiv e in g re d ie n t in th e p r o d u c t." EPA s a id f a ilu r e to re q u e s t a h e a rin g " w ill n o t p re v e n t t h e d is tr ib u tio n o r s a le o f e x is tin g s to c k s o f c e r t a i n p ro d u c ts w hose r e g is tr a tio n s a r e c a n c e l le d . T h e a g e n c y w ill p e rm it th e c o n tin u e d d is tr ib u tio n o r s a le o f e x is tin g s to c k s o f 2 ,4 ,5 - T a n d silv ex p r o d u c ts w h ich a r e la b e le d f o r e n d u s e s s u b je c t t o th is n o tic e , a n d w h o se r e g i s t r a t i o n s a r e c a n c e lle d p u rs u a n t t o th is n o t ic e , fo r n o m o re th a n o n e y e a r fro m th e e f f e c ti v e d a t e o f s u c h c a n c e lla tio n . H o w e v e r, th is p ro v isio n w ill a p p ly o n ly to th o s e e x is tin g s to c k s o f 2 ,4 ,5 - T a n d silv ex p r o d ­ u c ts w h ic h w e r e p a c k a g e d a n d la b e le d fo r su c h e n d u s e s a n d r e le a s e d fo r s h ip m e n t p rio r t o th e d a te o f r e c e ip t o f th is n o tic e by th e r e g i s tr a n t o r p u b lic a tio n o f th is n o tic e in th e F e d e ra l R e g is te r , w h ic h e v er o c c u rs f i r s t . T h e a g e n c y w ill n o t p e r m it th e c o n tin u e d d is tr ib u tio n , s a le , o r u se o f a n y c a n c e lle d 2 ,4 ,5 - T o r silv e x p r o d u c t la b e le d f o r a n y e n d u s e s u b je c t t o th e 1979 e m e r g e n c y su s p e n s io n o r d e r o r fo r m a n u f a c tu r in g u s e ." T h e a g e n c y 's l e t t e r t o r e g is tr a n ts c o n tin u e d : "E P A n o te s th a t m any r e g is tr a n ts rec e iv in g th is l e t t e r m a y hold r e g i s tr a ti o n s fo r p ro d u c ts w h ich w e re o rig in a lly la b e le d fo r su sp e n d e d u s e s o f 2 ,4 ,5 - T o r silv e x . R e g is tr a n ts o f p r o d u c ts w h ic h w e r e la b e le d fo r o n e or m o re su sp e n d e d u s e s a n d w h o e le c t n o t t o p a r t i c i p a t e in ' a n y f u r th e r h e a rin g c o n c e rn in g su c h p r o d u c ts m ay m e e t th e q u a li f ic a ­ tio n s fo r rec e iv in g in d e m n if ic a tio n p a y m e n t u n d e r F IF R A S e c tio n 1 5 ( a ) ." T h e a g e n c y a ls o n o te d p ro v isio n fo r EPA d isp o sa l o f th e c a n c e lle d p e s tic id e s . to th e r e g is tr a n ts sig n e d by E dw in L . Jo h n so n , OPP D i r e c t o r , s t a t e d : " T h e tim e -c o n s u m in g a n d c o s tly n a tu r e o f c o n tin u e d lit ig a t io n s u p p o r ts m y b e lie f t h a t it w ill be in th e p u b lic i n t e r e s t if th e 2 ,4 ,5 - T /s ilv e x c a n c e lla tio n p ro c e e d in g c a n b e c o n c lu d e d w i t h ­ o u t th e ta k in g of a d d itio n a l te s tim o n y . T he le tte r c " I f th e h e a rin g re s u m e s , it is e s tim a te d th a t it w ill ta k e a t le a s t o n e y e a r to b rin g th e a d m in is tra tiv e p ro c e e d ig to c o n c lu s io n , a llo w in g fo r s u p p le m e n ta l p r e - t r i a l a c tiv itie s in c lu d in g d isc o v e ry , p r e s e n t a t io n o f a d d itio n a l te s tim o n y , b r ie f in g , a n d iss u a n c e of .re c o m m e n d e d a n d f in a l d e c is io n s , p lu s a n o th e r y e a r o r m o re t o re s o lv e ju d ic ia l a p p e a ls . " W h ile m u ch o f th e e v id e n c e of risk s a s s o c ia te d w ith u se of 2 ,4 ,5 - T a n d silv ex h a s b e e n p r e s e n te d , EPA a n t i c i p a t e s t h a t a d d itio n a l ris k te s tim o n y w ould have t o be p r e s e n te d c o n c e r n ­ ing in f o r m a tio n w h ic h h a s a c c u m u la te d d u rin g th e tw o an d o n e - h a l f y e a r p e rio d s in c e th e h e a rin g w as s u s p e n d e d . " In a d d itio n , s in c e th e b e n e f its p r e s e n ta tio n h a d b a r e ly g o t te n u n d e rw a y w h e n t h e h e a rin g w a s r e c e s s e d , v itu a lly t h e e n ti r e b e n e f its c a s e w ill have t o .b e p r e s e n te d ." T h e a g e n c y 's S e c tio n 6 ( b ) ( 1 ) c a n c e lla tio n n o tic e s a id , " I f a r e g i s tr a n t w hose 2 ,4 ,5 - T o r silv e x p ro d u c t is la b e le d fo r m o re th a n one o f th e u s e s s u b je c t t o th is n o tic e o r o t h e r _ a d v e rs e ly a f f e c t e d p e r s o n e l e c t s t o re q u e s t a h e a rin g c o n c e rn in g s o m e b u t n o t a ll a f ­ f e c t e d u s e s o f t h e p r o d u c t , th e r e g is tr a n t a ls o m u st s u b m it t o th e R e g is tr a tio n D iv isio n , O PP, a n a p p li c a t i o n t o a m e n d th e r e g is tr a tio n o f s u c h p r o d u c t to d e le t e a ll o th e r a f f e c t e d u s e s w ith in t h e s a m e 3 0 - d a y p e rio d w ith in w h ic h a h e a rin g m ay b e r e q u e s te d . F a ilu r e t o s u b m it s u c h a n a p p lic a tio n fo r a m e n d e d r e g i s tr a ti o n w ill r e s u lt in c a n c e lla tio n o f th e e n t i r e r e g i s tr a ti o n o f th e p ro d u c t by o p e ra tio n of la w ." It c o n tin u e d : , " i f a r e g i s tr a n t o r o th e r a d v e rs e ly a f f e c te d p e rs o n r e q u e s ts a h e a rin g p u rs u a n t t o th is n o tic e fo r y 2 ,4 ,5 - T or silv e x p r o d u c t fo r w h ich no h e a rin g h a s p re v io u sly b e e n r e q u e s te d a n d t h e l a b e l­ in g s u b m itte d fo r t h a t p r o d u c t h a s n o t b e e n p rev io u sly a m e n d e d t o d e le te a ll u se s w h ic h w e re s u b je c t t o th e e m e r g e n c y s u s p e n s io n a n d S e c tio n 6 ( b )( 1 ) c a n c e l la t io n n o tic e s fo r 2 ,4 ,5 - T a n d silv e x issu e d in 1 9 7 9 , t h e r e g i s tr a n t a ls o m u st s u b m it t o th e R e g is tr a tio n D ivision of th e O ffic e o f P e s tic id e P ro g ra m s a n a p p lic a tio n to a m e n d su c h la b e lin g t o d e l e t e su c h p re v io u sly c a n c e lle d u s e s w ith in th e s a m e 3 0 - d a y p e rio d w ith in w h ich a h e a rin g m ay b e r e q u e s te d . F a ilu r e t o s u b m it s u c h a n a p p lic a tio n fo r a m e n d e d r e g is tr a tio n w ill r e s u lt in c a n c e l la t io n of th e e n ti r e r e g i s tr a ti o n o f th e p e s tic id e . " C o a s e q u e n c e s of f a ilu r e t o f ile in a tim e ly a n d e f f e c tiv e m a n n e r . If no h e a rin g h a s b e e n r e q u e s te d w ith in th e a p p lic a b le 3 0 -d a y p e rio d fo r a n y 2 ,4 ,5 - T or silvex p ro d u c t w h ic h is la b e le d f o r o n e o r m o re u s e s s u b je c t t o th is n o tic e an d th e r e g i s tr a n t h a s n o t s u b m itte d t o EPA a n a p p li c a t i o n t o a m e n d th e r e g i s tr a ti o n of su c h p ro d u c t t o d e l e t e a ll u se s s u b je c t t o th is n o tic e , t h e e n t i r e r e g is tr a tio n o f th e p r o d u c t w ill be c a n c e lle d . H o w e v e r, if a r e g is tr a n t d o e s not re q u e s t a h e a rin g fo r a 2 ,4 ,5 - T o r silv ex p ro d u c t w h ich is la b e le d f o r o n e o r m o re u s e s s u b je c t t o th is n o t ic e , b u t d id p re v io u s ly r e q u e s t a h e a rin g c o n c e rn in g c a n c e l la t io n of o n e o r m o re u s e s of th a t p r o d u c t w h ic h w e r e s u b je c t t o th e e m e rg e n c y su sp e n sio n n o tic e s a n d n o tic e o f in te n t t o c a n c e l 2 ,4 ,5 - T a n d silv ex issu e d in 1 9 7 9 , th e r e g is tr a n t h a s th e rig h t to c o n tin u e to p a r t i c i p a t e in a h e a rin g w ith r e s p e c t t o t h e l a t t e r u s e s . At th e c o n c lu s io n o f th e 2 ,4 ,5 -T /s ilv e x c a n c e l la t io n p r o c e e d in g , r e i n s ta t e m e n t o f s u c h u se s w ill be g r a n te d t o t h e e x te n t p e r m itte d by th e fin a l a d ju d ic a to r y o r d e r . A ny r e g i s tr a n t or fo rm e r r e g is tr a n t w h o r e q u e s ts r e i n s ta t e m e n t o f a r e g i s t r a t i o n fo r s u c h u s e s p u rs u a n t to su c h a fin al a d ju d ic a to r y o r d e r w ill n o t be re q u ire d to c o m p ly w ith S e c tio n 3 ( c ) ( 1 ) ( D ) of F1FRA in o rd e r fo r su c h r e i n s ta t e m e n t t o be g r a n te d ." V D ow , in e n d in g its p a r t i c i p a ti o n in th e h e a rin g a n d w ith d ra w in g its h e a rin g r e q u e s t, s a id , " 2 ,4 ,5 - T a n d silv ex a r e s a f e , e f f e c tiv e h e rb ic id e s t h a t n e it h e r p o se nor t h r e a te n u n r e a s o n ­ a b le a d v e rs e e f f e c t s o n th e e n v iro n m e n t a n d a r e t h e r e f o r e p ro p e rly r e g is te r e d u n d e r th e F IF R A . S c ie n tif ic p a n e ls , r e g u la to ry a u th o r itie s a n d ju d ic ia l d e c is io n m a k e rs in t h is c o u n tr y a n d th ro u g h o u t th e w o rld , in clu d in g E P A 's ow n S c ie n tif ic A dvisory P a n e l in re v ie w in g p a r ts of th is p r o c e e d in g , h a v e r e a c h e d s im ila r c o n c lu s io n s ." ( 4911 'V-sooCo T h e c o m p a n y f u r t h e r s t a t e d , " in ta k in g th is a c tio n , Dow is h o p e fu l th a t re s o u rc e s h e r e to f o r e d e v o te d t o th is p ro c e e d in g w ill be fre e d fo r m ore c o n s tr u c tiv e u se in a d d re s s in g th e b ro a d e r is s u e s o f th e p r e s e n c e o f c h e m ic a ls , including d io x in , in th e e n v iro n m e n t- F u r th e r p u rs u it o f th is p r o c e e d in g c o u ld d e t r a c t fro m t h a t b ro a d e r in q u iry ." J o h n s o n , re sp o n d in g t o D o w 's re q u e s t fo r v o lu n ta ry c a n c e lla tio n s , a c c e p te d th e r e q u e s t, e f f e c ti v e im m e d ia te ly . He s a id , " e x is tin g s to c k s of s u c h p ro d u c ts w h ich w e re f o r m u la te d , p a c k a g e d , la b e le d o n ly fo r e n d u se s w hich have not p rev io u sly b e e n su sp e n d e d , a n d re le a s e d f o r s h ip m e n t by y o u r firm on o r b e fo re O c t. 14, 1 9 83, m ay be d is tr ib u te d an d so ld u n til O c t. 1 4 , 1 9 8 4 , b u t n o t t h e r e a f t e r ." T h e a g e n c y o f f ic ia l c o n c lu d e d , " w h ile we d o n o t a g r e e w ith a ll o f th e s t a t e m e n ts c o n c e r n - , in g 2 ,4 ,5 - T a n d silv ex . . ., w e d o a g re e th a t it is in th e p u b lic i n te r e s t to b rin g th e 2 ,4 ,5 - T /s ilv e x p ro c e e d in g to a c lo s e ." In a n O c t. 14 l e t t e r t o A lvin L . A im , EPA D e p u ty A d m in is tr a to r , D o w 's K .R . M cK ennon, G ro u p V ic e P r e s id e n t, s a id , " f o r tu n a t e ly , p ro g re s s m ad e by Dow a n d o th e r s in h e rb ic id e p r o d u c t d e v e lo p m e n t s in c e 1979 h a s a ls o r e s u lte d in s u b s ti tu t e p ro d u c ts b ein g a v a ila b le f o r m o st u s e s o f 2 ,4 ,5 - T a n d silv ex . Dow is c o n v in c e d t h a t th e s e s u b s titu te s a r e now s u f f i c i e n tl y a v a ila b le t o s a tis f y its o b lig a tio n t o c u s to m e r s ." H e c o n tin u e d : " W ith t h e iss u e o f 2 ,4 ,5 - T a n d silv ex b e h in d u s , Dow h o p e s t h a t it c a n h e lp t o fo rg e a la s tin g p a rtn e rs h ip w ith EPA a n d th e m any s c i e n t i f i c g r o u p s , e n v iro n m e n ta l o rg a n iz a tio n s , a n d in d u s tr ia l c o m p a n ie s h av in g a n i n t e r e s t a n d e x p e r tis e in d io x in m a t te r s ." T h e c a n c e l l a t i o n n o tic e a n d e n f o r c e m e n t p o lic y w e re p u b lish e d in th e O c t. 18 F e d e ra l R e g is te r . 4912 'J J - S b o 7 t" * Occupational Health Guideline for 2,4,5-T * INTRODUCTION This guideline is intended as a source of information for employees, employers, physicians, industrial hygienists, and other occupational health professionals who may have a need for such information. It does not attempt to present all data; rather, it presents pertinent information and data in summary form. SU B S T A N C E IDENTIFICATION • Formula: C lsC«H iO CH aCO O H • Synonyms: 2,4,5-Trichlorophenoxyacetic acid • Appearance and odor: Colorless to tan odorless solid. P ER M ISS IB LE EX P O S U R E LIMIT (PEL) T he current OSHA standard for 2,4,5-T is 10 milligrams of 2,4,5-T per cubic meter o f air (mg/m*) averaged over an eight-hour work shift. HEALTH HAZARD INFORMATION • Routes of exposure 2,4,5-T can affect the body if it is inhaled or if it comes in contact with the eyes or skin. It can also affect the body if it is swallowed. • Effects of overexposure Exposure to 2,4,5-T may cause abdominal pain, nausea, vomiting, diarrhea, and blood in the stool. It may also cause irritation o f the skin. Common contaminants of commercial preparations o f 2,4,5-T may cause acne and liver damage. Animal experiments have shown that these contaminants may produce damage in unborn rats. • Reporting signs and symptoms: A physician should be contacted if anyone develops any signs or symptoms and suspects that they are caused by exposure to 2,4,5-T. • Recommended medical surveillance The following medical procedures should be made available to each employee who is exposed to 2,4,5-T at potentially hazardous levels: 1. Initial Medical Examination: —A complete history and physical examination: The purpose is to detect pre-existing conditions that might place the exposed employee at increased risk, and to establish a baseline for future health monitoring. Exami­ nation o f the liver and attention to gastrointestinal complaints should be stressed. The skin should be examined for evidence o f chronic disorders. 2. Periodic Medical Examination: The aforementioned medical examinations should be repeated on an annual basis. • Summary of toxicology 2.4.5- T (2,4,5-trichlorophenoxyacetic acid) is o f low toxicity. T he oral LD50 for dogs is in the range of 100 m g A g o r higher; effects are limited to a slight or m oderate stiffness in the hind legs with development of ataxia. Contaminants o f commercial preparations o f 2.4.5- T have been 2,3,7,8-tetrachlorodibenzo-p-dioxin, a potent animal teratogen, and 2,3,6,7-tetrachlorodibenzo-p-dioxin (TCD D), a potent acnegenic agent which is hepatotoxic in animals; they are present as unwanted side products of synthesis o f 2,4,5-T. In a study o f 73 workers in a 2,4,5-T manufacturing plant, 13 had m oderate to severe acneform dermatitis (chloracne) and 22 had gastrointestinal complaints such as nausea, vomiting, diarrhea, abdominal pain, or blood in the stool; no significant liver dysfunction was found; al­ though no air sample results w ere reported, the chlor­ acne was thought to be a result of exposure to TC D D . 2.4.5- T dust is a slight irritant o f the skin. C H EM IC A L AND P H Y S IC A L P R O P E R T IE S • Physical data 1. M olecular weight: 255.5 2. Boiling point (760 mm Hg): Decomposes above melting point 3. Specific gravity (water = 1): G reater than 1 4. V apor density (air = 1 at boiling point o f 2,4,5-T): Not applicable 5. Melting point: 158 C (316 F) (decomposition) These recommendations reflect good industrial hygiene and medical surveillance practices and their implementation will assist in achieving an effective occupational health program. However, they may not be sufficient to achieve compliance with all requirements of OSHA regulations. UJS. DEPARTMENT OF HEALTH AND HUMAN SERVICES Public Health Service Centers for Disease Control National Institute for Occupational Safety and Health September 1978 U.S. DEPARTM ENT O F LABO R Occupational Safety and Health Administration 491? 6. Vapor pressure at 20 C (68 F): Essentially zero 7. Solubility in water, g/100 g water at 20 C (68 F): 0.03 8. Evaporation rate (butyl acetate = 1): Not applica­ ble • Reactivity 1. Conditions contributing to instability: Tem pera­ tures above 158 C (316 F) may cause sealed metal containers to burst. 2. Incompatibilities: None. 3. Hazardous decomposition products: Toxic gases and vapors (such as hydrogen chloride and carbon monoxide) may be released when 2,4,5-T decomposes. 4. Special precautions: None. • Flammability 1. Not combustible • Warning properties 2,4,5-T is not known to be an eye irritant. MONITORING AND M EASUREM EN T P R O C ED U R ES • General Measurements to determine employee exposure are best taken so that the average eight-hour exposure is based on a single eight-hour sample or on two four-hour samples. Several short-time interval samples (up to 30 minutes) may also be used to determine the average exposure level. A ir samples should be taken in the employee’s breathing zone (air that would most nearly represent that inhaled by the employee). • Method An analytical method for 2.4.5-T is in the N IO SH Manual o f Analytical Methods. 2nd Ed., Vol. 5, 1979, available from the Government Printing Office, W ash­ ington, D.C. 20402 (G PO No. 017-033-00349-1). R E S P IR A T O R S • Good industrial hygiene practices recommend that engineering controls be used to reduce environmental concentrations to the permissible exposure level. H ow ­ ever, there are some exceptions where respirators may be used to control exposure. Respirators may be used when engineering and work practice controls are not technically feasible, when such controls are in the process of being installed, o r when they fail and need to be supplemented. Respirators may also be used for operations w hich require entry into tanks or closed vessel's, and in emergency situations. If the use o f respirators is necessary, the only respirators permitted are those that have been' approved by the Mine Safety and Health Administration (formerly Mining Enforce­ ment and Safety Administration) or by the National Institute for Occupational Safety and Health. • In addition to respirator selection, a complete respira­ tory protection program should be instituted which includes regular training, maintenance, inspection. 2 2.4.5-T cleaning, and evaluation. SANITATION • Eating and smoking should not be permitted in areas where 2 ,4 ,5 ^ is handled, processed, or stored. • Employees who handle 7,4,5-T should wash their hands thoroughly with soap o r mild detergent and water before eating, smoking, o r using toilet facilities. COMMON O PERA TIO N S AND CO N TRO LS T he following list includes some common operations in which exposure to 2,4,5-T may occur and control methods which may be effective in each case: Operation Controls Formulation of herbicides and plant hormones P ro cess enclosure; local exhaust ventilation; personal protective equipment Application as herbicide, defoliant, and plant hormone Personal protective equipment Manufacture of 2,4,5-T P ro cess enclosure; local exhaust ventilation; personal protective equipment - EM ER G EN C Y F IR S T AID P R O C ED U R ES In the event o f an emergency, institute first aid proce­ dures and send for first aid or medical assistance. * Eye Exposure If 2,4,5-T gets into the eyes, wash eyes immediately with large amounts of water, lifting the lower and upper lids occasionally. If irritation is present after washing, get medical attention. Contact lenses should not be worn when working with this chemical. • Skin Exposure If 2,4,5-T or liquids containing 2,4,5-T get on the skin, wash the contaminated skin using soap or mild deter­ gent and water. If 2,4,5-T or liquids containing 2,4,5-T soak through the clothing, remove the clothing and wash the skin using soap or mild detergent and water. If irritation is present after washing, get medical attention. • Breathing If a person breathes in large amounts o f 2,4,5-T, move the exposed person to fresh air at once. If breathing has stopped, perform artificial respiration. Keep the affect­ ed person warm and at rest. G et medical attention as soon as possible. * Swallowing When 2,4,5-T o r liquids containing 2,4;5-T have been swallowed and the person is conscious, give the person large quantities o f water immediately. After the water has been swallowed, try to get the person to vomit by having him touch the back o f his throat with his finger. D o not make an unconscious person vomit. Get medical 4914 S ep tem b er 1978 attention immediately. • Rescue Move the affected person from the hazardous exposure. If the exposed person has been overcome, notify some­ one else and put into effect the established emergency .escue procedures. D o *iot 'become a casualty. U nder­ stand the facility’s emergency rescue procedures and know the locations of rescue equipment before the need arises. S P IL L AND D ISP O SA L P R O C ED U R ES • Persons not wearing protective equipment and cloth­ ing should be restricted from areas o f spills until cleanup has been completed. • If 2,4,5-T is spilled, the following steps should be taken: 1. Ventilate area of spill. 2. Collect spilled material in the most convenient and safe manner and deposit in sealed containers for recla­ mation, or for disposal in a secured sanitary landfill. Liquid containing 2,4,5-T should be absorbed in vermiculite, dry sand, earth, or a similar material. • Waste disposal method: 2,4,5-T may be disposed of in sealed containers in a secured sanitary landfill. REFEREN CES • American Conference of Governmental Industrial lygienists: “2,4,5-T (2,4,3-Trichlorophenoxacetic Acid),” Documentation o f the Threshold Lim it Valuesfo r Substances in Workroom Air (3rd ed., 2nd printing), Cincinnati, 1974. • Christensen, H. E., and Luginbyhl, T. L. (eds.): N IO SH Toxic Substances List, 1974 Edition, HEW Publication No. 74-134, 1974. • Deichmann, W. B., and Gerarde, H. W.: Toxicology o f Drugs and Chemicals, Academic Press, New York, 1969. • Drill, V. A., and Hiratzka, T.: “Toxicity of 2,4Dichlorophenoxyacetic Acid and 2,4,5Trichlorophenoxyacetic Acid,” A.M.A. Archives o f In­ dustrial Hygiene and Occupational Medicine, 7:61-67, 1953. • International Labour Office: Encyclopedia o f Occupa­ tional Health and Safety, McGraw-Hill, New York, 1971. • Khera, K. S., and McKinley, W. P.: “Pre- and PostNatal Studies on 2,4,5-Trichlorophenoxyacetic Acid, 2,4-Dichlorophenoxyacetic Acid and Their Derivatives in Rats,” Toxicology and Applied Pharmacology, 22:14-28, 1972. • Patty, F. A. (ed.): Toxicology, Vol. II of Industrial Hygiene and Toxicology (2nd ed. rev.), Interscience, N ew York, 1963. • Poland, A. P., et al.: “A Health Survey o f Workers in a 2,4-D and 2,4,5-T Plant,” Archives o f Environmental Health, 22:316-327,1971. • Spencer, E. Y.: Guide to the Chemicals Used in Crop Protection (6th ed.), Publication 1093, Research Branch Agriculture, Canada, 1973. • S P E C IA L NOTE T he International Agency for Research on Cancer (IARC) has evaluated the data on this chemical and has concluded that it causes cancer. See I A R C Monographs on the Evaluation o f Carcinogenic Risk o f Chemicals to Man, Volume 15, 1977. 4915 S o \o S ep tem b er 1978 2,4,5-T 3 g . 2 ,3 ,7 ,8-TETR ACHLORODIBENZO-p-DIOXIN K Agency for Toxic Substances and Disease Registry U.S. Public Health Service ^ ^ t, i I €> o <0 ° DRAFT TOXICOLOGICAL PROFILE FOR 2,3,7,8-TETRACHLORODIBENZO-p-DIOXIN Date Published — December 1987 Prepared by: Syracuse Research Corporation under Contract No. 68-03-3228 for Agency for Toxic Substances and Disease Registry (ATSDR) U.S. Public Health Service in collaboration with U.S. Environmental Protection Agency (EPA) Published by: Oak Ridge National Laboratory under DOE Interagency Agreement No. 1425-1425-Al DISCLAIMER Mention of company name or product does not constitute endorsement by the Agency for Toxic Substances and Disease Registry. 0 FOREWORD The S u p e r fu n d Am endments a n d R e a u t h o r i z a t i o n A c t o f 1986 ( P u b li c Law 9 9 -4 9 9 ) e x te n d e d a n d am ended t h e C o m p re h e n siv e E n v iro n m e n ta l R e s p o n s e , C o m p e n s a tio n , a n d L i a b i l i t y A c t o f 1980 (CERCLA o r S u p e r f u n d ) . T h is p u b l i c law ( a l s o known a s SARA) d i r e c t e d t h e A gency f o r T o x ic S u b s ta n c e s a n d D is e a s e R e g i s t r y (ATSDR) t o p r e p a r e t o x i c o l o g i c a l p r o f i l e s f o r h a z a r d o u s s u b s t a n c e s w h ic h a r e m o st commonly fo u n d a t f a c i l i t i e s o n t h e CERCLA N a t i o n a l P r i o r i t i e s L i s t a n d w h ic h p o s e t h e m o st s i g n i f i c a n t p o t e n t i a l t h r e a t t o hum an h e a l t h , a s d e te r m i n e d by ATSDR a n d t h e E n v iro n m e n ta l P r o t e c t i o n A gency (E P A ). The l i s t o f t h e 100 m o st s i g n i f i c a n t h a z a r d o u s s u b s t a n c e s w as p u b l i s h e d i n t h e F e d e r a l R e g i s t e r on A p r i l 1 7 , 1 9 8 7 . S e c t i o n 110 (3 ) o f SARA d i r e c t s t h e A d m i n i s t r a t o r o f ATSDR to p r e p a r e a t o x i c o l o g i c a l p r o f i l e f o r e a c h s u b s t a n c e on t h e l i s t . Each p r o f i l e m u st i n c l u d e t h e f o l l o w i n g c o n t e n t : "(A ) An e x a m i n a t i o n , sum m ary, a n d i n t e r p r e t a t i o n o f a v a i l a b l e t o x i c o l o g i c a l i n f o r m a t i o n a n d e p i d e m io l o g ic e v a l u a t i o n s on th e h a z a r d o u s s u b s ta n c e i n o r d e r t o a s c e r t a i n th e l e v e l s o f s i g n i f i c a n t human e x p o s u r e f o r t h e s u b s t a n c e a n d t h e a s s o c i a t e d a c u t e , s u b a c u te , and c h ro n ic h e a l t h e f f e c t s , (B) A d e t e r m i n a t i o n o f w h e th e r a d e q u a te i n f o r m a t i o n on t h e h e a l t h e f f e c t s o f e a c h s u b s ta n c e i s a v a i l a b l e o r i n t h e p r o c e s s o f d e v e lo p m e n t t o d e te r m in e l e v e l s o f e x p o s u r e w h ic h p r e s e n t a s i g n i f i c a n t r i s k t o hum an h e a l t h o f a c u t e , s u b a c u t e , a n d c h r o n i c ____ h e a l t h e f f e c t s , and (C) W here a p p r o p r i a t e , a n i d e n t i f i c a t i o n o f t o x i c o l o g i c a l t e s t i n g n e e d e d t o i d e n t i f y t h e t y p e s o r l e v e l s o f e x p o s u r e t h a t may p r e s e n t s i g n i f i c a n t r i s k o f a d v e r s e h e a l t h e f f e c t s i n h u m a n s ." T h is t o x i c o l o g i c a l p r o f i l e i s p r e p a r e d i n a c c o r d a n c e w i t h g u i d e l i n e s d e v e lo p e d b y ATSDR a n d EPA. The g u i d e l i n e s w e re p u b l i s h e d i n th e F e d e r a l R e g i s t e r on A p r i l 1 7 , 1 9 8 7 . E ach p r o f i l e w i l l b e r e v i s e d and r e p u b l i s h e d a s n e c e s s a r y , b u t no l e s s o f t e n t h a n e v e r y t h r e e y e a r s , a s r e q u i r e d by SARA. The ATSDR t o x i c o l o g i c a l p r o f i l e i s i n t e n d e d t o c h a r a c t e r i z e s u c c in c tly th e t o x ic o lo g i c a l and h e a l t h e f f e c t s in fo r m a tio n f o r th e h a z a r d o u s s u b s t a n c e b e i n g d e s c r i b e d . E ach p r o f i l e i d e n t i f i e s a n d r e v ie w s t h e k e y l i t e r a t u r e t h a t d e s c r i b e s a h a z a r d o u s s u b s t a n c e 's t o x i c o l o g i c a l p r o p e r t i e s . O th e r l i t e r a t u r e i s p r e s e n t e d b u t d e s c r i b e d i n l e s s d e t a i l t h a n t h e k e y s t u d i e s . The p r o f i l e i s n o t i n t e n d e d t o b e a n e x h a u s t i v e d o c u m e n t; h o w e v e r, m ore c o m p re h e n s iv e s o u r c e s o f s p e c i a l t y i n f o r m a t i o n a re re fe re n c e d . iii E ach t o x i c o l o g i c a l p r o f i l e b e g in s w i t h a p u b l i c h e a l t h s t a t e m e n t , w h ich d e s c r i b e s i n n o n t e c h n i c a l la n g u a g e a s u b s t a n c e 's r e l e v a n t t o x i c o l o g i c a l p r o p e r t i e s . F o llo w in g t h e s t a t e m e n t i s m a t e r i a l t h a t p r e s e n t s l e v e l s o f s i g n i f i c a n t human e x p o s u r e a n d , w h ere known, s i g n i f i c a n t h e a l t h e f f e c t s . The a d e q u a c y o f i n f o r m a t i o n t o d e te r m in e a s u b s t a n c e 's h e a l t h e f f e c t s i s d e s c r i b e d i n a h e a l t h e f f e c t s sum m ary. R e s e a rc h g ap s i n t o x i c o l o g i c a n d h e a l t h e f f e c t s i n f o r m a t i o n a r e d e s c r i b e d i n t h e p r o f i l e . R e s e a rc h g a p s t h a t a r e o f s i g n i f i c a n c e t o p r o t e c t i o n o f p u b l i c h e a l t h w i l l b e i d e n t i f i e d by ATSDR, t h e N a t i o n a l T o x ic o lo g y P ro g ra m o f t h e P u b l ic H e a l th S e r v i c e , a n d EPA. The f o c u s o f t h e p r o f i l e s i s on h e a l t h a n d t o x i c o l o g i c a l i n f o r m a t i o n ; t h e r e f o r e , we h a v e i n c l u d e d t h i s i n f o r m a t i o n i n t h e f r o n t o f t h e d o c u m e n t. The p r i n c i p a l a u d ie n c e s f o r t h e t o x i c o l o g i c a l p r o f i l e s a r e h e a l t h p r o fe s s io n a ls a t th e f e d e r a l, s t a t e , and lo c a l le v e l s , i n te r e s te d p r i v a t e s e c t o r o r g a n i z a t i o n s a n d g r o u p s , a n d m em bers o f t h e p u b l i c . We p l a n t o r e v i s e t h e s e d o c u m e n ts i n r e s p o n s e t o p u b l i c com m ents a n d a s a d d i t i o n a l d a t a becom e a v a i l a b l e ; t h e r e f o r e , we e n c o u ra g e comm ent t h a t w i l l make t h e t o x i c o l o g i c a l p r o f i l e s e r i e s o f t h e g r e a t e s t u s e . T h is p r o f i l e r e f l e c t s o u r a s s e s s m e n t o f a l l r e l e v a n t t o x i c o l o g i c a l t e s t i n g a n d i n f o r m a t i o n t h a t h a s b e e n p e e r r e v ie w e d . I t h a s b e e n r e v ie w e d b y s c i e n t i s t s fro m ATSDR, EPA, t h e C e n t e r s f o r D is e a s e C o n t r o l , and t h e N a t i o n a l T o x ic o lo g y P ro g ra m . I t h a s a l s o b e e n r e v ie w e d b y a p a n e l o f n o n g o v e rn m e n t p e e r r e v i e w e r s a n d w as made a v a i l a b l e f o r p u b l i c r e v ie w . F i n a l r e s p o n s i b i l i t y f o r t h e c o n t e n t s a n d v ie w s e x p r e s s e d i n t h i s t o x i c o l o g i c a l p r o f i l e r e s i d e s w i t h ATSDR. Ja m e s 0 . M ason, M .D ., D r. P .H . A s s i s t a n t S u rg e o n G e n e r a l A d m i n i s t r a t o r , ATSDR iv CONTENTS FOREWORD ......................................................................................................................................... iii LIST OF FIGURES ............................................................................. v ii LIST OF TABLES ........................................................................................................................... ix 1. 2. PUBLIC HEALTH STATEMENT ......................................................................................... 1 .1 WHAT IS DIOXIN? ............................................................................................... 1 .2 HOW MIGHT I BE EXPOSED TO 2 , 3 , 7 , 8-TCDD? ....................................... 1 .3 HOW DOES 2 , 3 , 7 , 8-TCDD GET INTO MY BODY? ....................................... 1 .4 HOW CAN 2 , 3 , 7 , 8-TCDD AFFECT MY HEALTH? ......................................... 1 .5 IS THERE A MEDICAL TEST'TO DETERMINE WHETHER I HAVE BEEN EXPOSED TO 2 , 3 , 7 , 8-TCDD? ............................................................... 1 .6 WHAT LEVELS OF EXPOSURE BY INGESTION AND BY SKIN CONTACT HAVE RESULTED IN HARMFUL HEALTH EFFECTS? .................................... 1 .7 WHAT RECOMMENDATIONS HAS THE FEDERAL GOVERNMENT MADE TO PROTECTHUMAN HEALTH? ................................................................ 1 1 1 2 2 HEALTH EFFECTS SUMMARY ............................................................ 2 .1 INTRODUCTION ................. 2 .2 LEVELS OF SIGNIFICANT EXPOSURE ............................................................. 2 . 2 . 1 Key S t u d i e s a n d G r a p h ic a l P r e s e n t a t i o n s ...................... 2 . 2 . 2 B i o l o g i c a l M o n ito r in g a s a M e asu re o f E x p o su re a n d E f f e c t s .................................. ....................................................... 2.2.3 E n v iro n m e n ta l L e v e ls a s I n d i c a t o r s o f E x p o s u re a n d E f f e c t s .................................... .................................................... 2 .3 ADEQUACY OF DATABASE ....................................................... 2 . 3 . 1 I n t r o d u c t i o n ............................................................ 2 . 3 . 2 A dequacy o f t h e D a ta b a s e f o r H e a l th E f f e c t End P o i n t s ............................................................................................ 2 . 3 . 3 A dequacy o f t h e D a ta b a s e f o r O th e r I n f o r m a t i o n N eeded f o r R is k A s s e s s m e n t ..................................................... 7 7 8 8 3 4 4 18 19 21 21” 21 25 3. CHEMICAL AND PHYSICAL INFORMATION ......................................... 3 .1 CHEMICAL IDENTITY .......................................................................................... 3 .2 PHYSICAL AND CHEMICAL PROPERTIES ...................................................... 29 29 29 4. TOXICOLOGICAL DATA .................................................................................................... 4 .1 OVERVIEW ................................................................................................................. 4 . 2 TOXICOKINETICS .................................................................................................... 4 . 2 . 1 A b s o r p tio n ............................................................................................ 4 . 2 . 2 D i s t r i b u t i o n ....................................................................................... 4 . 2 . 3 M e ta b o lis m ............................................................................................ 4 . 2 . 4 E x c r e t i o n ............................................................................................... 4 .3 TOXICITY .................................................................. 4.3.1 L e t h a l i t y a n d D e c r e a s e d L o n g e v ity ..................................... 4.3.2 S y s t e m i c / T a r g e t O rgan T o x i c i t y ...................... ..................... 4 . 3 . 3 D e v e lo p m e n ta l T o x i c i t y ................................................................ 33 33 37 37 38 39 40 41 41 42 49 4920 v 4 . 3 . 4 R e p r o d u c t i v i t y T o x i c i t y ............................................................. 4 . 3 . 5 G e n o t o x i c i t y ....................................................................................... 4 . 3 . 6 C a r c i n o g e n i c i t y ................................................................................ INTERACTIONS WITH OTHER CHEMICALS ........................ ............................. 50 53 56 60 MANUFACTURE, IMPORT, USE, AND DISPOSAL ...................................................... 5 .1 OVERVIEW ................................................................................................................ 5 .2 PRODUCTION ............................................................................................................. 5 .3 IMPORT ....................................................................................................................... 5 .4 USE ....................................................................................... ...................................... 5 .5 DISPOSAL .................................................................................. 61 61 61 61 61 61 ENVIRONMENTAL FATE .................................................................................................... 6 .1 OVERVIEW ....................................................................................... 6 .2 RELEASES TOTHE ENVIRONMENT ......................................................... 6 . 2 . 1 P r o d u c t i o n a n d Use o f C e r t a i n H e r b i c i d e s an d C h lo r o p h e n o ls ........................................................................... 6 . 2 . 2 P h o to c h e m ic a l R e a c t i o n s ............................................................. 6 . 2 . 3 T h e rm a l R e a c ti o n s ...................................... 6 . 2 . 4 Im p ro p e r D i s p o s a l o f C h l o r i n a t e d C h e m ic a l W a ste s . 6 .3 ENVIRONMENTAL FATE .......................................................................................... 63 63 63 7. POTENTIAL FOR HUMAN EXPOSURE ..................................................... 7 . 1‘* OVERVIEW .................................................. 7 . 2 . LEVELS MONITORED OR ESTIMATED IN THE ENVIRONMENT ................... 7 . 2 . 1 A i r ........................................................................ 7 . 2 . 2 W a te r ...................................... 7 . 2 . 3 S o i l ..................................... 7 . 2 . 4 O th e r ........................ 7 .3 OCCUPATIONAL EXPOSURES ........................ 7 .4 POPULATIONS AT HIGH RISK .......... ..................... .......................................... 67 67 68 68 68 69 69 73 73 8. ANALYTICAL METHODS ........................ 8 .1 ENVIRONMENTAL MEDIA ................................................... .. . .............................. 8 . 1 . 1 A i r , W a te r, S o i l , a n d F o o d ....................................................... 8 .2 BIOMEDICAL SAMPLES ............ 8 . 2 . 1 F l u i d s / E x u d a t e s a n d T i s s u e s ...................................................... 75 76 76 76 76 9. REGULATORY AND ADVISORY STATUS ........................................................................ 9 .1 INTERNATIONAL (WORLD HEALTHORGANIZATION) ..................................... 9 .2 NATIONAL............................. 9 . 2 . 1 R e g u l a t i o n s ............................................................ 9 . 2 . 2 A d v is o ry G u id a n c e ............................................................................ 9 . 2 . 3 D a ta A n a l y s is ...................................................................... 9 .3 STATE .......................................................................................................................... 83 83 83 83 83 83 84 10. REFERENCES .......................................................................................................................... 85 11. GLOSSARY ...................................................................... 109 4 .4 5. 6. _ APPENDIXES A. PEER REVIEW .................................................................................. B. FEDERAL REGISTER ANNOUNCEMENT .................................................................. 63 64 64 64 65 115 117 LIST OF FIGURES 1 .1 H e a l th e f f e c t s from i n g e s t i n g 2 , 3 , 7 , 8 - T CD D ......................................... 5 1.2 H e a l th e f f e c t s from s k i n c o n t a c t w i t h 2 , 3 , 7 , 8 - T CDD ...................... 6 2.1 E f f e c t s o f 2 , 3 , 7 , 8- TCDD- or a l e x p o s u re ........................................................ 10 2 .2 E f f e c t s o f 2 ,3 ,7 ,8 -T C D D -d e rm a l e x p o s u re 11 2 .3 L e v e ls o f s i g n i f i c a n t e x p o s u r e f o r 2 , 3 , 7 , 8- TCDD- or al 2.4 L e v e ls o f s i g n i f i c a n t e x p o su re f o r 2, 3, 7, 8-TCDD-dermal ............. 13 2.5 A dequacy o f t h e d a ta b a s e on h e a l t h e f f e c t s o f 2 , 3 , 7 , 8 - T CD D (hum an d a t a ) .................................................................................................................... 22 A dequacy o f t h e d a ta b a s e on h e a l t h e f f e c t s o f 2 , 3 , 7 , 8 - T CDD ( a n im a l d a t a ) ................................................................................................................. 23 2.6 ................................................... ................. 12 4921 p - 5 09-° Or .“ó LIST OF TABLES 3.1 Chemical identity 3.2 Physical properties 4.1 Recommended TEFs 4.2 Genotoxicity of 2,3,7,8-TCDD 4.3 Genotoxicity of 2,3,7,8-TCDD 4.4 Summary of et (1978a,b) 4.5 al. Other oral the oral is 2,3,7,8-TCDD of for .................................... 2,3,7,8-TCDD 2,3,7,8-TCDD .......... and ............ 36 in v i t r o ............................... 54 in v i v o ................................ 55 carcinogenicity Its an supporting animal 2,3,7,8-TCDD the congeners bioassay conclusion carcinogen 7.1 Levels 8.1 -Analytical met h o d s for 8.2 Analytical for b i o m e d i c a l methods in 30 31 of Kociba ........................................................... studies 2,3,7,8-TCDD of of soil from environmental that ................................ differentlocations 58 ........ 70 ................. 77 ......................... 80 samples samples 57 (5 ix y .b 0 1. 1.1 WHAT The PUBLIC chemical 2,3,7,8-tetrachlorodibenzo-p-dioxin called name 2 , 3 , 7 , 8 - T C D D . It for dioxin. 2,3,7,8-TCDD is Dioxin, is standard. and It during present the time, scientific 1.2 can be during colorless known to is an solid with no occur except inadvertently produced in v e r y incineration of not as nor a used and for purpose it amounts and industrial any is reference small certain herbicides of municipal is is colloquial distinguishable naturally industry, manufacture ( 2 ,3,7,8-TCDD) inaccurate any 2,3,7,8-TCDD as an germicides, w a s t e s . At other the than research. _MOW MIGHT The the however, a neither intentionally manufactured b y impurity STATEMENT IS D I O X I N ? commonly odor. HEALTH main I BE TO environmental • Production acids EXPOSED and use 2,3,7,8-TCDD? sources of of h e r b i c i d e s 2,3,7,8-TCDD containing are: 2,4,5-trichlorophenoxy (2,4,5-T) • Production and use of • Production and of h e x a c h l o r o p h e n e • Incineration • Small use 2,4,5-trichlorophenol of municipal amounts formed and during certain as a germicide industrial the b u r n i n g of w o o d wastes in the presence of chlorine • Accidental benzenes • Exhaust transformer/capacitor from • Improper reported. than The the of one sections but million of it times several powered with certain level involving of soil of chlorinated than in Lake Ontario, Saginaw European fish not bee n methods countries. trace in Missouri, detected sensitive at in the soils in u r b a n from normal Bay, the in of measurement, It h as levels 2,3,7,8-TCDD not been the is n o t not rural from and air has in m o s t obtained detected wastes in a m bient in d r i n k i n g w a t e r 2,3,7,8-TCDD was gasoline chemical 2,3,7,8-TCDD present higher leaded chlorinated 2,3,7,8-TCDD can be 2,3,7,8-TCDD has using more of concentrations oil-contaminated 2,3,7,8-TCDD milk, on Detection undetectable, waste automobiles disposal Information available. fires and biphenyls been soils is soils. level urban was areas. contaminated Michigan the U n i t e d rivers. States; it h a s been detected in In human however, detected any other 4923 i In a more in foods. 2 Consumer sources: • Skin contact with by chemical the • Consumption • Inhalation municipal of of surfaces root such as vegetables air n e a r soil grown or v e g e t a t i o n in contaminated contaminated improperly maintained dump soils sites or • incinerators • Consumption of fish, livestock, and cow's milk from contaminated areas • Breast-fed babies nursed by mothers who have 2,3,7,8-TCDD in their breast milk • Since both 2,4,5-T-containing herbicides been withdrawn sources occur of from c o n s u m e r use, exposure; however, from hazardous Occupational this exposures longer to involved in the hexachlorophene, chemical. — ■- h o w e v e r , has The been production and production of and • Workers at certain hazardous waste • Workers involved • Workers involved in cleanup fires and of other and industrial DOES and containing 2,4,5-trichlorophenol, States. incinerators of certain in the accidental salvaging spraying phenoxy herbicides 2,3,7,8-TCDD • Ingestion through of cow's • Breathing milk, and of transformers such as A g e n t body fly however, may • Intake of should be CAN through the soils and consumption other of materials contaminated ambient intake. air may constitute 2,3,7,8-TCDD from contribute Inhalation the of a major source consumption very small particulates of of such as exposure. drinking water negligible. 2,3,7,8-TCDD occurs chloracne initial contaminated 2,3,7,8-TCDD AFFECT MY In humans, usually BODY7 foodstuffs contaminated total INTO M Y from 2,3,7,8-TCDD to ash, GET skin amounts HOW trichlorophenol . • Absorption 1.4 may sites Orange fish, have future compounds herbicides the U n i t e d certain municipal the use or 2,4,5-T in at in or 2,4,5-T • Workers HOW be sites. discontinued capacitor/transformer 1.3 these will sources: • Workers salts, waste and hexachlorophene they no is on more the causes head HEALTH? chloracne, and upper disfiguring and body. often a severe Unlike lasts for skin lesion common acne, years after that the exposure. AQ 3 • There is suggestive evidence i n h u m a n s , as indicated by in the Animal in some • There blood. is suggestive weight exposure to prior to death. • Although causes in l e v e l s of liver damage certain studies have demonstrated severe liver evidence that 2,3,7,8-TCDD causes loss never loss, and 2,3,7,8-TCDD result in greater • Although never exposure to digestive results demonstrated 2.3.7.8- TCDD produced monkey 2,3,7,8-TCDD increase enzymes damage species. appetite, can that an in severe in humans, toxicity to demonstrated sensitive immune to in humans, resulted to this • Although never demonstrated 2.3.7.8- TCDD during pregnancy loss of body weight just studies, system. This toxicity infection. in some animal ih spontaneous toxic property in h umans, of A n im a l in humans. in a nimal the susceptibility 2,3,7,8-TCDD is v e r y disorders of The 2,3,7,8-TCDD. in rodents, resulted species, abortions. exposure in malformations to in the offspring. • 2,3,7,8-TCDD has been demonstrated to be a carcinogen 1 .5 _^IS T H E R E A M E D I C A L T E S T T O D E T E R M I N E W H E T H E R I BEEN EXPOSED There Is n o unequivocally that be a blood helpful not common medical test since to these were 2.3.7.8- TCDD fat b y presence serum is initial detect a other tests simple of higher the test exposure you have as demonstrate indicating occurred. been exposed only have in It to can of the damage These than available, One to cause consists for the that average than in m o t h e r ' s milk would 2.3.7.8- in may was to b a c k g r o u n d not milk fat presence the of levels or if been used widely to evaluate of If the also some additional human In you have been a piece small then analyzed sensitive levels in of to addition, for blood The to 2,3,7,8-TCDD States, more detection exposure; indication of enough the U n ited exposed exposure test, 2,3,7,8-TCDD. is indicate conducted that developed for people population. provide is method probably been 2.3.7.8- TCDD has in suspected commonly removing recently another 2.3.7.8- TCDD the of the you have average not In 2,3,7,8-TCDD. determined that the do similar changes people indicate procedure; indicate of whi c h ' a r e adequately test analyzed levels the may tests 2,3,7,8-TCDD, been performed, some is b e l i e v e d liver has drinking alcohol, to m o r e surgical appears indicates TCDD to 2,3,7,8-TCDD. enzymes enzymes 2,3,7,8-TCDD. background are to ________ appear obtained and study well these 2,3,7,8-TCDD. the HAVE exposure. are body that demonstrated a physician but to as for available exposed certain certainty tests enzymes exposed detect chemicals, When There test been in d e t e r m i n i n g w h e t h e r other animals. 2,3,7,8-TCDD? that yo u have indicate with results. by TO in the of whether occurred. This of level of exposure method exposure. 4925 4 1.6 WHAT LEVELS HAVE RESULTED The graphs exposure graphs to exposure side EXPOSURE on the "Health is m e a s u r e d In effects the effects products The of of on the o n Fig. cancer, day would noted estimates!. lower. that Actual There was these risk (One n a n o g r a m not in o f 10,000 the are or some of 1.1), of body the left relationship to b e are of associated with are based on Protection Agency (EPA) per additional cases people, to b e has 2,3,7,8-TCDD plausible a in still of is with uncertainty 1,560,000 unlikely information on contact in humans, 10,000,000 one-billionth enough again anticipated risk values levels is 1560 same skin 1 nanogram or kilogram shown (mg/kg). Environmental to per are measured therefore, exposure result in a p o p u l a t i o n should be ~ the U.S. lifetime set (Fi g . are cancer exists. between first of body weight as studies; the 2,3,7,8-TCDD" from than animal cancer 1.2), 1.1, from In side. effects information kilogram per right 2 , 3 , 7 , 8 - T C D D ." E x p o s u r e s marked CONTACT relationship in animals (Fi g . other that the 2,3,7,8-TCDD "Health effects For of graphs known SKIN effects. ingesting effects for estimated show 2,3,7,8-TCDD per kilogram levels risk from graphs, set the containing milligrams minimal for pages in millig r a m s BY EFFECTS? and known health in humans second represented INGESTION AND following (mg/kg). In all and BY IN H A R MFUL HEALTH 2,3,7,8-TCDD labeled weight OF of respectively. It upper-limit higher and may be gram.) to p r e p a r e a graph for exposure by — breathing. 1.7 WHAT RECOMMENDATIONS PROTECT Both (IARC) the have suggests The HUMAN EPA and concluded Agency for 2,3,7,8-TCDD causes cancer 2,3,7,8-TCDD may EPA calculated health effects expected are not (mg/L) 0.00000001 0.0000001 mg/L 0.035 ppt, (0.035 is, for The ambient water that fish serious EPA also and and the rivers) of takes exposure fish. may The less concerns. lifetime calculated additional of HA ppt) the 1,000,000 into of occur cancer ppt of which 0.000001 a child; an of that health milligrams the 10-day HA longer-term HA for in adverse adult mg/L is (0.1 and for adults 2,3,7,8-TCDD in associated with over background 0.000000000013 through both Drug Cancer 2,3,7,8-TCDD 0.000000035 amount to b e account Federal 25 is which would be incidence than HA is for mg/L containing health 1-day trillion, 0.000000035 calculated measurement hence, The child in a p o p u l a t i o n eating to occur. on in animals, below which a incidence the levels per Research (HAs). f o r of The (lakes fish; advisories a child. in one in TO in humans. for increases and cancer (0.001 ppt) respectively); ppt). cause estimates (10 p a r t s mg/L MADE International that liter FEDERAL GOVERNMENT the that is THE that d r i n k i n g water, per HAS HEALTH? 2,3,7,8*-T C D D the drinking Administration has 2,3,7,8-TCDD cancer mg/L. This concentrates of water calculated should pose no 5 LONG-TERM EXPOSURE (GREATER THAN 14 DAYS) SHORT-TERM EXPOSURE (LESS THAN OR EQUAL TO 14 DAYS) E FF EC T S IN ANIMALS D O SE (mg/kg/day) EFFECTS IN HUMANS 0.001 EFFEC TS IN ANIMALS DEATH DOSE (mg/kg/day) EFFEC TS IN HUMANS 0.00001 DEATH. 0.0005 0.000005 REPRODUCTIVE I TOXICITY AND Ä CHLO RACN E________ 0.000001 0.0001 0.00008 DEVELOPMENTAL < E FF EC T S X 0.0000008 T 0.00004 0.0000006 LIVER DAMAGE < 0.00002 1 0.0000004 T 0.000008 i, T 0.0000002 0.000006 0.0000001 1 0.000004 T 0.00000005 0.000002 1 T 0.000001. MINIMAL RISK FOR E FF EC T S OTHER THAN . CANCER 0.00000001 T 0.000000005 0.000000001 MINIMAL RISK FOR EFFECTS OTHER THAN CANCER Fig. 1.1. Health effects from ingesting 2,3,7,8-TCDD. f 4927 SHORT-TERM EX P O SU R E (L E S S THAN OR EQ U A L TO 14 DAYS) EFFECTS IN ANIMALS D O SE (mg/kg/day) EFFECTS IN HUMANS LONG-TERM EX P O S U R E (G R EA T ER THAN 14 DAYS) EFFECTS IN ANIMALS DOSE (mg/kg/day) QUANTITATIVE DATA W ERE NOT AVAILABLE QUANTITATIVE DATA W ER E NOT AVAILABLE 0.01 0.8 0.008 0.6 0.006 0.4 0.004 DEATH EFFECTS IN HUMANS C H L O R A C N E __________ 0.2 0.002 0 0 Fig. 1.2. Health effects from skin contact with 2,3,7,8-TCDD. 2. 2.1 This present section summarizes exposure levels toxicological exposure Potential The the and The differ media of develop such effects. on about their of the the exposure user's may be (Frank of want on animals Level, posing minimal risk health professionals on displays fqrmat these data 2.2. Toxic Toxicology a and of Program (NTP) agenda have no been levels Risk graphs highlights in of the health at in graphs this section information to will the exposed officials more sites subtle Level, (No-Observed- Estimates are to associated— Superfund effects areas Registry U.S. to p r o v i d e of of levels interest 7 availability be literature consistent is to establish with suggest levels to Health and Service information. of key and the where of considered by ( A T S D R ) , EPA, Public this the scientific These Disease may potential exposure Levels) in physicians findings of fluids alike. 2.3) purpose adverse (2) information associated with observed. (Minimal (Sect. of a found the the Public three-dimensional research with and in h u m a n example, actions in and groups exposures. shown on clinical exposure and includes with and health associated with FEL). 2,3,7,8-TCDD The public response to exposure. Substances Level, citizens insufficient human in parts. (Lowest-Observed-Adverse-Effect NOAEL) Database in Sect. significant 7, narratives 2,3,7,8-TCDD For overt below which to h u m a n s exposure t^iere m i g h t b e develop of Effect levels levels Adverse -Effect Adequacy Sect. 2,3,7,8-TCDD also levels interested may or brief levels of persons concerned with information of of perspective. frank effects studies to critically two m a j o r individuals section levels identification disease exposure and association with human interpretation the in huma n s is on key environmental Data, provides exposure This project managers or based is that have be e n monitored and effects effects section section presents toxicology with LOAEL) this comprises interested the significant and significance the this Toxicological section other of information or w i t h in that and concerned with of investigations, and in a m anner 2,3,7,8-TCDD persons the h e a l t h for 2,3,7,8-TCDD 2.2) and health depending on purpose (Sect. of environmental 4, Summary Exposure studies depiction tissues, exposure Sect. perspective adverse data The Exposure. physicians, levels in Effects for key summarized graphs information presented discussed (l)^an overall various significant Significant officials, and 2,3,7,8-TCDD. studies, epidemiological Health of graphics to for Human This Levels of data. evaluated and for SUMMARY INTRODUCTION concerning on HEALTH EFFECTS the the in Agency National order to 8 2.2 LEVELS To living OF help SIGNIFICANT public or w o r k i n g summarized in health points this that section are systemic/target toxicity, in terms Two a kinds animal of route does reflect second kind and LOAELs the reflect No were versus shows actual adjustments actual end points. human These starting from the established derive are low actual The of levels exposure) of or The Significant of the for animal the graph (levels of exposure) or exist level It risks (level uncertainty and of were are dose used In Studies is between (10*^ to on risk are pig sensitive. TCDD. In soil, which It addition, may compound. on of the graphs exposure animal and methods have (Barnes graphs the most applying a broken ending with etal. 1987), confidence, in the cancer EPA. In addition, tumor to assess are many apparently is n o t the end point incidence the . is the effect from Animal studies administered differences remainder of in to the data most have are on to h u m a n s exposure and the hamster humans are to sensitive adsorbs toxicity; to sensitive, toxicity monkey is the and such as toxicity of conducted with from which data the 2,3,7,8- to materials the b i o a v a i l a b i l i t y insufficient the from in generally been should be presented. = differences strongly affect 2,3,7,8-TCDD. sensitivity the risk in oily vehicles of bioavailability exposure the species 2,3,7,8-TCDD — ~ Presentations known how significantly a been under by data estimates as exposure) reduce reported for the associated with Graphical readily bioavailable. There risk and key for n o n c ancer duration by levels that made derived shown the 10'^) exposure) and and guinea each experimental considered when Although techniques shown difficult 2.3.7.8- TCDD .<0 ° the extrapolating exposure (level terminus. Also were each levels its the of There this The for showing NOAELs intermittent differences levels these minimal in 2 . 3 . 7 . 8 - TCDD. least effects ■ ;: Key is and tested. (LSE) plotted. 2.2.1 type human each of Exposure on discussed chronic. first ordering species duration are and data. points for actual at estimates. dose the to h u m a n h e a l t h w e r e risk These line projected reproductive data summary conditions end point concave-curved to the risk minimal factors. shortcomings of The graphical available. variations, exposure noncancer uncertainty toxicity, toxicological intermediate, depict a exposure reflecting posing minimal "sensitive general made. intraspecies of_levels then by areas--lethality, duration doses data of six (and are The for route dermal--and to Levels duration. the Adjustments to man, data exposure graph exposure studies. end points of persons by carcinogenicity. used needs toxicology first periods--acute, are the the developmental and It p r ovides for wh i c h of protocol line graphs toxicological not toxicity, graph. exposure route and into exposure address sites, organized ingestion, toxicity, three "thermometer" are waste categorized organ genetic of professionals near hazardous exposure--inhalation, end EXPOSURE the compound available studies used Both bioavailability considered when to to and is consider define species evaluating the the 9 Oral graphs data and dermal in Figs. are NOAELs 2.1 available and for graphically present Levels for the of oral inhalation dermal exposure inhalation data for graphical for oral exposure three-generation observed fetal in relatively the fetuses. exposure, which short these are "thermometer" qualitative toxicity as level data were depicted insufficient the The study effects well for in Figs. insufficient calculated by chronologically risk on some representation. Since duration, same minimal presented Although There were was reproductive the are to effects. routes. risk level LOAELs respectively. h u m a n s , quantitative significant and and 2.2, in as the 2.4 and animal (1985a) the rat, an chronic exposure with effects to exposure exposure was minimal from a attributed represents chronic and intermediate EPA were 2.3 human in u t e r o of of the calculated dams, from data. 2.2.1.1 Inhalation No studies 2 . 3 . 7 . 8- TCDD. occurred to are available Exposure the in accidental population releases on through or the exposed in inhalation inhalation, to toxicity however, chemicals the w o r k p l a c e of may also have contaminated with TCDD (especially herbicide spraying). 2.2.1.2 Oral Lethality death and in h u m a n s 2,3,7,8-TCDD there is from 0.6 a large /ig/kg i n m a l e and the to for 18 feeding after days in approximately pigs are Target of observed Because of also of some to b e of al. 1986), 2.1 and are hyperkeratosis, exposed female and dermal, a 2.1 occurs 9 0 - d a y ______ (0.008 occur that the /ig/kg mortality 9 months Four major hirsutism loss, chemicals some although in did not and though 5500 usually 50% monkeys for chloracne, changes, contaminant, of range in Figs. at ingested died (Allen NOAEL toxic in et guinea effects the w a s t i n g of the digestive and skin, industrial oral of libido and chemicals routé possible aching disorders, contaminated with the p r i m a r y some loss syndrome, immunotoxicity, and hypercholesterolemia, weight to to exposure and monkeys toxicity. sensory Death estimated i m m u n otoxicity. Chloracne, appetite, 1973) plotted deaths even L D 50 v a l u e s 2 , 3 , 7 ,8-TCDD/kg whereas reports 2.3 . hypertriglycerid a al. are Extended in a n no species, exposure. 0 . 8 /ig o f in guinea pigs herbicides as oral of eight 2,3,7,8-TCDD in humans et /ig o f 2 , 3 , 7 , 8 - T C D D / k g / d a y FELs insomnia, 2.3.7.8- TCDD likely total et organ/systemic loss acute a in Figs. hepatotoxicity, (Schwetz resulted hyperpigmentation, neuropathy, of been 2,3,7,8 -TCDD. sensitivity. These values exposure. hepatotoxicity, and muscles, 1981). have to to a l l m a m m a l i a n single 0.01 characteristic toxic There exposure in species (NOAEL). Five These shown oral guinea pigs (DeCaprio ^g/kg/day 1977). a longevity. of guinea pigs al . lethality consumption al. et after study ft g / k g / d a y ) 0.0006 result is h i g h l y (Henck 13 a difference in hamsters 2.3 decreased as of headaches, have been 2 , 3 , 7 , 8 -TCDD. contain exposure inhalation is m o s t exposure probably occurs. 4931 £>.-!? 0 3 ? - ANIMALS HUNWNS (yg/fcg/day) 10.000 (— 1,000 • HAM STER ID » SINGLE DOSE • MOUSE. MONKEY. CHLORACNE. SINGLE DOSE - 100 - K - 1 — • 0.1 — 0.01 - 0.001 - • MOUSE. DEVELOPMENTAL TOXICrTY. 10 DAYS. CONTINUOUS GUINEA PIG . LO *. SINGLE DOSE O MOUSE. DEVELOPMENTAL TOXICITY, 10 DAYS. CONTINUOUS • • RAT. DEVELOPMENTAL TO XICITY. 10 DAYS. CONTINUOUS GUINEA PIG. LIV ER TO XICITY. SINGLE DOSE • O GUINEA PIG. IMMUNOTOXtCITY, B W EEKS. INTERMITTENT RAT. DEVELOPMENTAL TO XICITY. 10 DAYS. CONTINUOUS • f* [O • MONKEY. DEATH. 9 MONTHS. CONTINUOUS: MONKEY, CHLORACNE. 7 MONTHS. CONTINUOUS GUINEA PIG. DEATH. 90 DAYS. CONTINUOUS GUINEA PIG . IMMUNOTOXICITY. 8 W EEKS. INTERMITTENT GUINEA PIG. WASTING SYNDROME. UVER TOXICITY. 90 DAYS. CONTINUOUS • MONKEY. REPRODUCTIVE TOXICITY, 7 MONTHS. CONTINUOUS • O O RAT. REPRODUCTIVE TO XICITY. 3 GENERATIONS: RAT. UVER TOXICITY. 2 YEARS, CONTINUOUS GUINEA PIG . WASTING SYNDROME, UVER TO XICITY. 90 DAYS. CONTINUOUS GUINEA PIG. DEATH. 90 DAYS. CONTINUOUS 0.0001 •— • LOAEL O NOAEL Fig. 2.1. Effects of 2,3,7,8-TCDD—oral exposure. QUANTITATIVE DATA WERE NOT AVAILABLE 11 ANIMALS HUMANS (ji^Vg/day) 1000 r- Q UAN TITATIVE DATA W ER E NOT A V A ILA BLE • R A B B IT. L D » SIN G LE DO SE • M O USE. D ERM AL LES IO N S . 4 W E E K S . IN TERM ITTEN T 100 10 - 1 *• LO AEL Fig. 2.2. Effects of 2,3,7,8-TCDD—dermal exposure. 4933 ACUTE (S14 D AYS) LETH A LITY D EVELO PM ENTAL CHRONIC (2365 DAYS) INTERM EDIATE (15-364 DAYS) TA RG ET D EC R EA SED ORGAN LO N GEVITY REPRODUCTION TARGET ORGAN TARGET ORGAN CAN CER (ugXg/day) 10.000 • s 1000 100 10 1 r r 0.1 0.01 » g (LIV ER ) • k g •k 0 . 001- • r g (BODY W EIG H T, LIV ER ) Ì (LIV ER ) •r - 0.0001 0.00001 0.000001 0.0000001 - 0.00000001 0.000000001 IO“* 0.0000000001 ESTIM A TED HUMAN CAN CER RISK 10"6 LEV ELS 10"5 - 0.00000000001 0.000000000001 10-7 - 0.0000000000001 * j MINIMAL R IS K LE V E L , FOR E F F E C T S O TH ER s i/ THAN C A N CER g GUIN EA PIG k M ONKEY m M O USE r RAT S H AM STER LO A EL AND NO AEL IN SAM E S P E C IE S I • LO A EL FO R ANIMALS O NOAEL FO R ANIMALS ' \ MINIMAL R IS K LEV EL FO R CHRONIC EXTRA PO LA TED FROM IN TERM EDIATE EXPO SU RE Fig. 2.3. Levels of significant exposure for 2,3,7,8-TCDD—oral ACUTE (£ 14 DAYS) INTERMEDIATE (15-364 DAYS) LETHALITY TARGET ORGAN CHRONIC (S 365 DAYS) (ng/kg/day) 10,000 r QUANTITATIVE DATA WERE NOT AVAILABLE • h 1000 - 100 - 10 • m (SKIN) 1 I- • LOAEL m MOUSE h RABBIT Fig. 2.4. Levels of significant exposure for 2,3,7,8-TCDD—dermal. 935 14 Since as species, mice, 70 on The in the in a face indicated this 7 -month a a et 2.1 . o nly be has been dose dose study al. The of 1978, limited. of a or al. a at dose of similar 1977). were are few In hairless produced studies FELs in 2,3,7,8-TCDD 70 Mg/kg, Allen et only in h u m a n s detected in monkeys, available relationship; identified can single single feeding (McConnell i n Fig. definitively effect after whereas a dose-response wasting syndrome is In acute studies, this dose-response a of lesion exposure, These not designed reported (see 2.1). weight. A only produced 1984), /ig/kg/day define Fig. was (Greig are the 2,3,7,8-TCDD investigation chloracne lesions data from the fig /kg -0.01 to chloroacne, resulting 90-day guinea relationship study pigs characterized by were syndrome for (DeCaprio diets extreme loss of body associated with the wasting in g u i n e a p igs maintained on is syndrome has et 1986). al. providing average lethal been doses. defined Female Hartley 2,3,7,8-TCDD 0, 0 . 1 2 , 0 . 6 8 , 4 . 8 6 , a n d 31 n g / k g / d a y . T h e h i g h d o s e a F E L ; a 40% d e c r e a s e i n b o d y w e i g h t a n d d e a t h o c c u r r e d . T h e 4 . 8 6 - n g / k g ( 0 . 0 0 5 p g / k g ) d o s e r e p r e s e n t e d a L O A E L , w i t h a 13% d e c r e a s e i n b o d y w e i g h t a n d n o m o r t a l i t y . T h e 0 . 6 8 n g / k g ( 0 . 0 0 0 7 p g / k g ) is a doses of represented NOAEL for this indicated in effect. Figs. 2,3-,"7,8 - T C D D severity damage of is the not -tested with 2.1 as to Collins In 20 (1983). /ig /kg . a a LOAEL available. A sensitive in (1986). The the There in are guinea pigs rats, no and et a dose of studies. Again, only available for The 0.001 this guinea a LOAEL, end point pig also decrease thymus weight, at doses also of groups of 0, produced 0.04 body in in f ig / k g weight of of 10 0.008, other and was which toxic total dose was et fig /kg same and 2.3). in 2.1 chronic studies "toxic dietary exposure these that 2 -year 2.1 sensitive (1973) number, high loss were and 2.3, species observed and total was of body comparable to controls. The LOAEL on Fig. but not liver and w a s t i n g syndrome are more dose defined. are p l o t t e d 2.1, al. in Figs. al. The including 0.008 et the 2,3,7,8-TCDD weekly ft g / k g . __ was ng/kg, Figs. in Fig. the is exposure. lymphocyte 0.2 damage reported lowest (see it w a s DeCaprio 4.86 however, immunotoxicity toxicity liver (see the m o s t given since guinea not exposure study by (1982a) Vos The a NOAEL was oral is p l o t t e d to b e effects of or low groups. syndrome chronic guinea pigs a NOAEL 12.5, 2,3,7,8-TCDD/kg/day after or 2.5, 0.1, LOAEL was the and 0.5, feeding at necrosis Turner at Similar and NTP focal by female but g u i n e a pigs; as species and acute the sensitive of male studies. 2,3,7,8-TCDD. 0.04, liver (1985a), and appears effects the Although EPA changes fig o f however, doses all 2.3), the w a s t i n g in in of 90-day (1978a,b) immunotoxic number the ng/kg degenerative provided and acute studies al. occurred LOAEL by for group are tested; such low exposure studied. the m o s t 2,3,7,8-TCDD this in species changes small 2.1 intermediate species at very effects in for pig, for 0.68 LOAEL chronic Kociba hepatitis" from all the liver a of Figs. risk NOAEL guinea liver end point and in on observed dose the NOAEL was NOAEL the study, (see calculated observed as this minimal most and lethality, been on considered was in single Effects 2.3) depends severe regard given 2. 3 . is h e p a t o t o x i c have was LOAEL and lesions and hypertrophy pigs The on the and NOAEL Fig. sensitive 2.3 end 8 weeks FEL weight. At the leukocyte for a to a and A LOAEL LOAEL, for because points of 936 15 intermediate to guinea oral pigs; exposures to exposure Thigpen et 2,3,7,8-TCDD to S a lm o n e lla -induced in animals not guinea (1975) at death. exposed Developmental developmental in al. Immunotoxicity reported doses as This dose low as is n o t limited that mice given 4 weekly 1 /ig/kg w e r e more sensitive caused no gross signs of toxicity S a lm o n e lla . to toxicity. toxicity pigs. There in humans have as a been result no of reports oral of exposure to 2,3,7,8-TCDD. 2.3.7.8- TCDD produces palate and hydronephrotic in rat. the FELs administration (Sparschu et in rats of al. the was a NOAEL fetal death. Although this species appears administered and Dillman 1972) "Tiurray toxicity levels dose et al. in that as a (1979) rats. doses of in d e c r e a s e d on size and fetal considered data, lowest dose weight, 0.001 and changes the In a risk abortions al. 2.3. 1979). This may toxicity provided FELs, NOAEL the as a dilated be and actual exposure TCDD has genotoxicity; however, suggest 2,3,7,8-TCDD that genotoxicity). experimental solubility of Some of the difficulties this is of a the reported sensitive been no are compound not 7 months, at b o t h indicated effects, regard to however, available for which there dose is species reports exposure frank study, that ( 1 9 8 5 a ) ______ pg/kg/day, species with This oral the fetal were levels on Figs. indicates the only determining a sensitivity. of genotoxicity in humans 2,3,7,8-TCDD. a testing the negative few positive genotoxic and level EPA the that 2.3). for monkeys observed 0.001-dose reevaluated 2.3). 0.01 severe data were to diet the indicated Fig. and (1979) decreased which at The high concluded chronic on diet effects that pelvises, (see inconsistencies in and and 0.0015 the al. (1982) produced mostly there et Paxton analysis reproductive reproductive (with (see have (when (Neubert /ig/kg/day. methods, index, relationship between There 2.3.7.8- and 2,3,7,8-TCDD. result oral Murray 0 . 0015-/ig/kg/day additional 0.1 and which Genotoxicity. of and of in intermediate two-thirds in mice, 2,3,7,8-TCDD. and in of to survival) maintained the most reports 2.1 of ft g / k g of -0.3 Figs. levels The observed a LOAEL renal (see (1982) 0.03 ^g/kg/day administered 0.01, gestational for tested, 1 /ig/kg c a u s e d three-generation survival. at study, that monkeys a Nisbet a LOAEL Paxton reproductive or the gestation also been no statistical study with monkeys et and and 2,3,7,8-TCDD spontaneous (Allen in fact a minimal Nisbet provided 2.1 in after of -1 exposure represented a NOAEL. different are FELs a NOAEL oral and neonatal tested produced /ig/kg w a s calculated from to b e using of cleft 2.3). 0.001, fetal 0 . 0 1 -dose of Doses and conducted 15 2.3). with including organ hemorrhage pg/kg/day dose 2,3,7,8-TCDD was the above of 0.125 lower effects and fetus, internal nçxt There have that was 2.1 result concluded litter and organogenesis) provided resulted 2.1 the 6 through the sensitive, Figs. to b e on days developmental toxicity. study and whereas Figs. less (see in humans in in mice reported compound (see during Reproductive toxicity were 1971a,b), Mg/kg, anomalies kidneys responses, Sect. 4.3.5 toxicity such as in v i v o 4937 in which tests may on observed may be 2,3,7,8-TCDD, high responses related the v e r y (which to low limits for 16 the quantity that can be tested), rather than to inherent biological inactivity. Carcinogenicity. incidence in h u m a n s There as 2,3,7,8-TCDD has both rats and mice a have result been et al. (1978a,b) total incidences derivation, palate, were et the or nasal combined. al . (1982a) from Kociba These data, Squire on Kociba et (1978a,b) The tumor 0, 0.001, al. the and whereas 27/48, q^* thus a that the cancer similarLy-v that for The tumor 1.56 x a expected cancer the Kociba similar used by et values ( 1 9 7 8 a , b), al. of data 0.01 exposure of risk would 0.0006 not a doses q^*. of 34/40, Squire were on Fig. cancer, to 2.3. The EPA has 0.6 1/10,000, pg/kg/day exceed derive and exposed exceed the pg/kg/day, indicated people to for from for reported by is 10,000 likely to 18/48, hard derived section (1978a,b) of data lung, 3/48, dose rat this liver, (1985a) al . 9/85, In in a 2 -year et set EPA in female q^*. ( m g / k g / d a y ) . For of is n o t lesser a incidence, a population risk used of carcinogen in rats the h i s t o l o g i c lower 10^ and derive tumors respective increased estimated the were 34/40. calculated was pg/kg/day, people, the an cancer 2 , 3 , 7 , 8-TCDD. (1985a) reported by along with study, and for animal reported by Kociba 16/85, associated with for increased to b e to 0. 1 /jg/kg/day w e r e respectively, 8/48, as study of to bioassay, réévaluation of incidences 0.01, reports exposure (1978a,b ) . EPA turbinates, EPA b y no oral demonstrated in an NTP bioassay by Kociba the been of and to 10,000,000 1/10,000,000 (Fig. 2. 3) . Lethality and death in humans Schwetz decreased as et a result As between application plotted o n Figs. in oral Target effect contaminated with been these most no information In on thus, addition, hepatotoxic data were Chloracne as it of data and as pointed porphyria is thus resulted cutanea in liver to pg/kg of time in LD50 The this that lesion. needed is to There to (EPA chloracne and in some assert injury, that and Chelsky of the the even if 2,3,7,8-TCDD also studies the studies, could (1986) , the presumed there and 2,3,7,8-TCDD is and other have been development liver diagnosis may be in however, cause exposure is however, performed. enzymes In all that is, to p r o d u c e 2,3,7,8-TCDD, 1985a). chemicals certain cannot be to h e r b i c i d e s liver Suskind have 2,3,7,8-TCDD that of and lesions exposed tarda tarda 275 (1979) body suggest out b y Jones difficult of only that serum levels been the In p o pulations contaminated with cutanea reports compounds from exposure assessment increased may have of length Taylor is b e l i e v e d exposure risk is and upper resulting and in producing levels are face chemicals exposure no available. certain reviewed by industrial of porphyria LD50 v a l u e dermal reports of been 2,3,7,8-TCDD. other As quantitative there to a protracted deforming compound in humans. a produced by compounds, the have was toxicity. in h u m a n s for many years effective and, No There exposure 2.4. 2,3,7,8-TCDD. aromatic the there death. and persistent, recognized halogenated humans, and 2.2 dermal reported studies, organ/systemic substantiated (1985), (1973) a l. rabbits. longevity. of damage, of questionable. to induced It 2,3,7,8-TCDD liver damage, i 17 there are no human data available that could provide dose-response information. The only animal data that are hairless mice given per for 4 weeks features of chloracne in h umans. the is 3.3 This week dose provided by provide chloracne a FEL; a dose-response for hence, 0.1 does have suggested effects Smith that et" a l . the and the effect. individuals any animal the studies of available dermal contaminated with al. McQueen 1978, a et 1979, Thomas Aldred 1978, Bisanti et 2.3.7.8- TCDD similar No to valid female is those animal 2.3.7.8- TCDD studies were following Carcinogenicity. epidemiology industrial studies indicated Hodgkins Hardell 1980, same studies lymphomas Lynge limitations questions studies for of industrial at 1985; as the populations data, exposure studies chemicals extent of 1979, of al. other Field New and exposure toxicity of of and Kerr 1979, 1980, Bonaccorsi studies or reproductive industrial Zealand 1982, These et did not adversely al. provide affects that exposure to the studies are of above. the reproductive et reviewed toxicity 1980; et exposures those with and of These as sites also a of well other of these development increase Hardell studies as of also where an et 1977; al. have the additional been grouped epidemiology contaminated herbicides above of in non- Frentzel-Beyme 1981; should have number and Some the and 1979, 1986). to an Theiss al. above, are series 2,3,7,8-TCDD. sites al. a to h e r b i c i d e s exposure of al. There to and 2,3,7,8-TCDD on of various with (EPA exposed described similar the human toxic reviewed human et also Eriksson Putoni analyses. chemicals et tumors 1977, indicated exposure. a variety those a l. of has section (1985a) 1979; et developmental limitations available (Axelson on whether The association Sandstrom 1981; together an to 2,3,7,8-TCDD 2,3,7,8-TCDD the performance, contaminated with sarcomas and that the dermal EPA exposed the 1980). of populations chemicals tissue effect. in 2.4 adversely in these of Health Smith al. indication discussed on and (1985a) limitations to h e r b i c i d e s reproductive without on Dept, 1980, or data 2,3,7,8-TCDD 1980, 1977, male 2.2 developmental EPA potentially (1985a) al. scientifically soft EPA Reggiani either the was defining exposure. exposed chemicals of McQueen 2,3,7,8-TCDD limitations were groups 1979, studies, of other group. Nelson et to quantitative toxicity. studies a variety these study following Reproductive toxicity major Figs. kg, which for contaminated with al. 0.03 dose exposure. an association with exposure specific within 2.3.7.8- TCDD The on times some weighs one of h u m a n populations a result prove three information dermal chemicals Nelson et as a mouse on in w h i c h resembled only used is p l o t t e d of inconsistent with but not concomitant 1-ffck o f No not could observed the Studies 1981, development, studies the were dose 1 9 8 2 ) .''"After r e v i e w i n g data were affecting these al. application necessary toxicity information a l . (1982), that that 2,3,7,8-TCDD produces et per however, the The organ et lesions Assuming study, industrial that (Hanify Puhvel dermal toxicity. other quantitative of 2,3,7,8-TCDD not provide target Developmental and of relationship. intermediate herbicides pg study developed pg/kg. it the elevated and incidence 4939 Ji-S 'cW o 18 of canc e r was 1980; al . Cook 1981; Wolfe et detected al. 1980, Johnson al. Besides not et the 1984; demonstrate an latency of with known the tumor lack exposure inconclusive data population. It w a s on the males, developed tumors ft g at 0.01 (NTP 1982b). There has tumor promotor. Poland mice, Berry not been but et able three also been mixed evidence not al . to (1982) observed Slaga demonstrate analysis of and the exposed could neither studies regarding females, the but not per week that 2,3,7,8-TCDD tumor-promoting for and Nesnows promoting the population the mice, times in mice heterozygous (1978), for application of application al. in to carcinogen. Swiss following, dermal 1983; additionally allow data animal In per et failed a control exposure et al. 1984). that exposure), a human 2 , 3 , 7 , 8-TCDD. al. et regarding to of the h u m a n is et indicated by from experimental of skin 2,3,7,8-TCDD period some of that Suskind Smith studies lack (as levels and and exposure were time 2,3,7,8-TCDD 2,3,7,8-TCDD data the population has actual a l . 1982; described apparent that some in hairless the Zack Pazderova-Vejlupkova Fingerhut cancer concluded carcinogenicity Similarly, 1985; again disprove are to et previously sufficient 1987; 1981; 2 , 3 , 7 , 8-TCDD, a general dermal have of 1980, Cook Pearce development, the There and of al. Riihimaki factors studies fat biopsies, nor 1981; Smith et 1986; association between confounded by prove al. confounding epidemiological no et (Ott 1985, the h a i r l e s s (1985), activity in is a activity trait. and NTP CD-I, (1982) Senear, or Swiss-Webster mice. 2.2.2. Biological There could are to b e one samples As adequate the extent indications of the have been elevated veterans exposed increased levels Agent et of in Missouri range of median 1986). times between transformer had median adipose 2.8 to 750 ppt, level of 6.4 and a Although six of the the than the Although the correlations available that in the obtained. failed tissues al . to (1984) detected of higher levels of reported in in Binghamton, 39 levels 20 was N e w York, ppt, with group had group also or individuals (Patterson exposed there 17 control ppt individuals exposed of _____ of Vietnam (1986), unexposed control, been levels study, subjects tissue of of individuals are with no tissue has adipose the b a c k g r o u n d generally an ubiquitous adipose exposure between In addition, 2.3.7.8- TCDD suggest the et shown tissue to h i g h accident to monitoring indication adipose adipose also b e e n 1.4 highest have (1983) al. been and that some exposed al . of has results 2,3,7,8-TCDD range tissue et Gross In another whereas samples however, et had a a al. levels extensive >5 overlap groups. qualitative exposure. fire Italy. of biological the whereas tissue have Seveso, higher in et Effects 2 , 3 , 7 , 8-TCDD, Young Nygren adipose a (1986), and Adipose for who had been in in sites Exposure 2,3,7,8-TCDD; obtained. Orange, 2.3.7.8- TCDD during to in vet e r a n s reported by accident analyses 2,3,7,8-TCDD to A g e n t of although mixed al. As the or exposure storage Orange. exposed a Measure tests of analyzed, levels as can be primary reported by Nygren detect / no indicate qualitative Monitoring may occurred, tissue level 5 there levels of and in the range to exposure to 2 , 3 , 7 , 8-TCDD, 18 of no the This makes good extent in of the exposure ppt. which some are 2,3,7,8-TCDD known history of provide to would it difficult 4940 19 to assess the additional exposure contribution exposure. to study to b o d y b u r d e n similar 2,3,7,8-TCDD a preliminary military A in and sera the M M W R histories indicating TCDD were compared 2.3.7.8- lack of levels (1987), exposure to 2,3,7,8-TCDD. In least, there was no difference the for based on lipid weight) the p r e s u m a b l y Biological provides possible available fluids, (Patterson et There with are is development and range 3.8 clear is chemicals for only the effects that in a n individual who occurred. The development extent exposure. of development of to 2,3,7,8-TCDD; indicated. In addition, is n o t with certainty exposure. studies (i.e., liver lipids) in 2.2.3 Environmental damage, determining Levels are significant permit the to be repeated elevate level little data exposure. use by areas to of and low half-life of has taken to b e into particular either effects safe and a lacked clear this single exposure or of of Exposure the of total hence, levels to to a exposure the level have that exposure would and calculated that substantially equivalent The areas of data Given of an to the the long individual levels misleading with been the 2,3,7,8-TCDD level. history of body burden. levels with in indicate could of not Effects studies environmental may be are in humans, would a high types and levels exposure 2,3,7,8-TCDD scenario environmental epidemiologic compound animal and been monitored (1986), exposure in and environmental 2,3,7,8-TCDD the insufficient occurred. association between Schlatter provide associated with in humans adequate not al. again, are observed contaminated with however, have account; observed areas levels 2,3,7,8-TCDD, single estimating a associate half-life the body burden of after Indicators 2,3,7,8-TCDD has accidents; Poiger exposure to et 2,3,7,8-TCDD has environment these obtained to in the biological 5 years exposure does there to this however, however, is to of Hoffman lipid metabolism, demonstrated found demonstration effects..The on as industrial of toxicity Levels human inhabitants of effects exposed exposure immunotoxicity would immunotoxicity signs not be e n that extensive herbicide through that Other have useful of data exposure TCDD There the for 2.3.7.8- 2.2.3.1 been that the exposure however, may have chloracne, Likewise, to associated c o m p o u n d s . The evidence of clearly aromatic that circulating serum of exposure is evidence state in human 2,3,7,8-TCDD; other halogenated the to in body effect had data only commonly parts- contaminated with caused by chloracne tests the indicate of group). detected 2,3,7,8-TCDD to ppt recent method with detected chemical extent is n o t With (1 (3.9 1987b). also suggest levels level nonexposed sensitivity has supportive (1986) the no at in b r e a s t milk, of exposure. 2,3,7,8-TCDD 2 , 3 , 2-f8-TCDD w o u l d p r o v i d e supportive for levels in with presumably of 2,3,7,8-TCDD a to reported results, 2,3,7,8-TCDD ppt indications with preliminary although also no of these monitoring techniques, Chloracne exposure chloracne as was containing or urine, al. TCDD. to h e r b i c i d e s the m e d i a n group such as b l o o d per-quadrillion in or qualitative analytical such 2.3.7.8- exposed monitoring, 2,3,7,8-TCDD non-Vietnam veterans unusual 9 ppt of small estimated in which V i e t n a m v eterans exposure to from any particular correlation between during regards to difficulties of discussed by Kimbrough a et 20 al. (1984), who concluded that >1 concern in r e s i d e n t i a l 2.2.3.2 Human exposure potential It is n o t possible to exposure with to 2,3,7,8-TCDD many human although there are each route. relative contribution route through direct formulations as a result of a herbicide the long soil or disposal. ingestion or In a 5-month in Mis s o u r i , area, pg/m^ level of may concentrate in. available mist of a also certain 2,3,7,8-TCDD, of liquid that containing occur chemical as a contaminant, in also areas occur from contaminated other removed with such a activities. from source fish, of from a between industrial 1 and and long-term, low- contaminated food which potentially Superfund a backhoe air were ingesting as or through 2,3,7,8-TCDD-contaminated represent sprayed adsorbed formed by the or by accidents concern where in of the 2,3,7,8-TCDD, may As routes a can industrial exposure levels providing of all quantify contamination) through 2,3,7,8-TCDD foods to 2,3,7,8-TCDD soil was Exposure that exposure half-life dust population. anticipated Dermal is b e i n g 1987). exposure. is of particular of human data the or the anticipated It 2,3,7,8-TCDD incinerators with no to is areas, soil -6 a l. 'also v a r i a b l e , of (soon after or du s t et inhalation route dermal. operation where average any p r e c i s i o n w h i c h contaminated contaminated (Fairless municipal or clean-up of containing foliage excavated site SOOO-ft^ is Inhalation may be are b e i n g levels environmental inhalation 2,3,7,8-TCDD. soils In are it contact with these herbicides waste 1.5 of exposure dermal of contact with with of 2,3,7,8-TCDD relevant scenarios, occur, of state with is m o s t exposure exposure significant ppb soil. are known is to relatively high exposures. Based on anticipated the adsorbed oil In to the Umbreit soil, as Missouri that of soils, of (1980) activated carbon, et al. strength of such' a s bioavailability It in is (1981) likely type that many also the h i g h through contaminated of the et al. administered 50% al . 2,3,7,8-TCDD that from Lucier et 1984, varies (1986b) for with soil was since bioavailability with contact (1982) contact time m a y less not in than be Poiger to type and and such as zero, and demonstrated time affect al. the Jersey may __ corn New available Philippi as in oily of strong binding vehicles, and is of also studies reduce of the such as result lipophilicity ingestion areas only it bioavailable soil. that Hence, the 2,3,7,8-TCDD. configuration, of and in bioavailability, that increases and animals, as from New Jersey limited and Huetter adsorption of et in be absorption but (McConnell apparently soil, exposure oral Bioavailability the v a r i a t i o n can not studies bioavailability to Because result 80%) demonstrated strong binding planar to s o i l s . The soil absorption substantial 1986a). where Schlatter factors available, demonstrated by Umbriet representative the 50 al. oral in experimental studies et and 2,3,7,8-TCDD will was. s t i l l from Missouri Philippi dermal used soil (absorption 1986, on adsorbed 2,3,7,8-TCDD vehicles. of data that is of nature physical and properties low water in v e r y h i g h fatty anticipated lipophilic same extremely bioconcentration long half-life tissues of which solubility fish to b e significant. of milk, secretion of and factors. 2,3,7,8-TCDD, that inhabit In addition, of result milk can as a provide f a o 21 relatively efficient 2,3,7,8-TCDD elimination exposures are in of of As the decreasing Since providing an both milk in a et can the fatty it form seems that of a l . (1986), milk and oily vehicle, 2,3,7,8-TCDD the b o d y b u r d e n Graham through mother's infant. would provide for discussed by 2,3,7,8-TCDD essentially sources mechanism females. result tissues likely is this in large of that fish these readily bioavailable. 2.3 ADEQUACY 2.3.1 OF DATABASE Introduct ion Section prepare a 110 (3) of SARA toxicological hazardous substances Priorities "(A) List. An found examination, hazardous £B) substance determination of significant risk Where health identifies levels identified of for levels areas such The Specific ATSDR, NTP, 2.3.2 Adequacy adequacy in h umans or in data, in the of and and in acute, on the of the effects. on the the h e a l t h process which of present subacute, a and for knowledge 2,3,7,8-TCDD. 2.2 of this relevant Such profile the ------- in and and mechanisms summarizes to gaps reproductive 2,3,7,8-TCDD, data in humans." (lethality, toxicity, for testing that may effects end points identifies for exposure health monitoring such toxicological of in c u r r e n t Sect. for of adequacy gaps, and for of of summarizes gaps. obtaining data needed 2,3,7,8-TCDD will be to develop developed by future. Database the levels health or levels exposure fill exposure and of developmental that may the the exposure adverse gaps section briefly animal Introduction The of human biological and EPA of types data reviewed significant of available evaluations information available levels risk research programs of adequate is content: of substance chronic identification the significant in progress levels 2.6, as and the effects present existing human an toxicity, of CERCLA National ascertain and certain health cancer) and for exposure toxicity, developing to to significant following epidemiologic significant organ points order of ATSDR 100 m o s t effects. significant section the to h u m a n h e a l t h identify system/target 2.3.2.1 in determine appropriate, to the interpretation and substance to developing research and of w h e t h e r each development present toxicity. summary, subacute, needed other include information acute, of f a c i l i t i e s -on t h e must exposure chronic are the A d m i n i s t r a t o r each associated effects This for significant human A (C) at Each profile toxicological “r* directs profile for graphic Health is End Points summary 2,3,7,8-TCDD animals Effect database depicted on bar for h e alth graphs in effect Figs. end 2.5 and respectively. 4943 ID .. Ò n O or HUMAN DATA a V ADEQUATE DATA J \ . SO M E >~ DATA J L E T H A L IT Y A C U TE ZI_______ IN TERM ED IA TE CHRONIC _______________________________________/ D EV ELO P M EN TA L R EPRO D U CTIVE TO X IC IT Y TO X IC ITY CARCIN O O EN ICITV S Y S T E M IC TO X IC ITY D CP Fig. 2.5. Adequacy of the database on health effects of 2,3,7,8-TCD D (human data). NO DATA N> ro A N IM A L D A T A A ADEQUATE V* DATA J SOME > J L E T H A L IT Y A C U TE IN TERM ED IA TE CHRONIC Z _______________________________________________________________/ D EV ELO PM EN TA L REPR O D U C TIV E TO X IC ITY TO X IC IT Y CA RCIN O G EN ICITY S Y S T E M IC TO X IC IT Y o S' CD Fig. 2.6. Adequacy o f the database on health effects of 2,3,7,8-TCD D (animal data). DATA NO DATA 24 The bars of full meet at least one of 1. For noncancer that meet a range cause 2. of There are health Bars exist not meet -clearly defined. the inhalation exposure; in contribute since humans were or This skin with soil Inhalation and inhalation of dust. reports repeated containing that are data regarding effects. fetal development, too limited mixed to a gaps to exposure compound on The the in of the a levels be a EPA cancer and not for the "adequate" to 2.5 the data cause this end to point data. for dermal studies occur these is not 2,3,7,8-TCDD, along with other routes are v e r y dermal will also to b e from areas chemicals occur likely ingestion (acute clear of on there an quantitative it is difficult. is after contact of the 2,3,7,8-TCDD. occur through contaminated food exposure as and well that chemicals also effects some for of association. indication however, that of 2,3,7,8-TCDD As of a on cancer result on populations anticipated there induction development data .. — exposures, considerably required the data data cpntaining hepatotoxic; the and dermal accidental exposure data to of The through indication are little h u m a n 2,3,7,8-TCDD. would disprove environment, 2,3,7,8-TCDD will a statistically does i n Fig. to the chloracne; or either by both f a c t o r . ~- contaminated with and is lack of a available there 2,3,7,8-TCDD prove that graphs human reproduction, the define (IARC) species, data contaminated were epidemiologic either and in dust cause containing these from of of human exposure 2,3,7,8-TCDD as derive exposures considered articles exposures one likely long-term exposure oral of that release contaminated to causes for the exposure are exposed other to bod y burden. of e x p o s u r e ) , there chemicals no and are indicates initial in exposure the studies to available levels carcinogen" Cancer "some" nature exposure effects to classified chemical indicated of highlights 2.5 from human From is the to on the that of Figure 2,3,7,8-TCDD. as of data oral data route. criteria Because toxicological most the instances, is human least that indicate substantially application show exposure and are sufficient (NOAELs) cancer potency of Descriptions obtained a exposure any of levels substance route some Human. the this adequacy only, 2.3.2.2 derive of half height Although the in at to "adequate" studies are sufficient a tumors studieis w h i c h exposure are carcinogenicity, effect via but_do data number sufficient and for Research (quantitative). of or more substance and animal one "probable (qualitative), are on a or potency signficant both carcinogen" the are FELs). International Agency For 4. or there standards from no-effect (LOAELs factor that conditions: points, carcinogenicity, "known human 3. end scientific toxicity effects indicate following health current For human and height the filling are of exposed data 25 There following are Animal. on the additional Figure although animal data, it specific species information species the on a must be toxicological dermal a the few studies studies 2 , 3 , ?"78-TCDD. likely to material This and a considerably more to species of oral is m o s t to studies. For may large data compared only apply differences these to are differences in L D 50 v a l u e s ) , c o m p l e t e for difficult to obtain. end point, the tested, it m u s t or most representative sensitivity is is a potentially data lethal a For the sensitive be clearly of humans. significant of of data on the the only this gap The in in Both dermal where the where is adsorbed chemical a al. 1973) two-stage types of of exposure to of are et these toxicity case the the of limited bioavailability studies route route (Schwetz 2,3,7,8-TCDD 2,3,7,8-TCDD which in dermal for in r a b b i t s the mous e . is p a r t i c u l a r l y significant available dose effects limited from are "adequate" adequately animal the and may have 2,3,7,8-TCDD large result species 2,3,7,8-TCDD in animals limited that toxicological (EPA 1985a) only occur a of 2,3,7,8-TCDD. of of there of magnitude exposure assay provide As species only determination tumorigenesis since a animal on that 2,3,7,8*TCDD end point will be of on toxicity exposure. primarily (4 o r d e r s data the recognized determined exposure, of are specific data Although and oral tested, understanding of adequate single data 2,3,7,8-TCDD. demonstrated which human on or indicates should be sensitivity complete data effects species observed with lack 2.6 toxicological humans, the no inhalation exposure in humans soil or is other compared with was applied in organic solvents. 2.3.2.3 A Summary review This individual is relevant of the studies. and studies coordinated by include carcinogenic soft tissue veterans 2.3.3 being are A birth of Database that extensive of for the to of U.S. Working particular are r e s e a r c h ----- 15 ongoing government Group. concern Subject for defects, and the studies are concerned with these for Other very enumerate ongoing consists Orange with show 2,3,7,8-TCDD area area the h e r b i c i d e A g e n t O r a n g e the of too conducted House Agent number is This and morbidity, anatomical sarcomas. of will important that effects, Adequacy a very (1985). the W h i t e to effort and Kang mortality exposed research However, reviewed by Young areas research toxicological properties considerable epidemiology ongoing F ederal R esearch in P ro g ress of investigations active. of development of in Vietnam. Information Needed for Risk Assessment 2.3.3.1 Pharmacokinetics Mechanisms under has been the There that is of is n e a r The the the that end points toxic locus mechanism As the action of functions this of properties for of reviewed b y 2,3,7,8-TCDD evidence toxicological segregation which action. investigation. proposed mechanism. of of extensive and mechanisms 2,3,7,8-TCDD Roberts by mechanism is as 2,3,7,8-TCDD 2,3,7,8-TCDD toxicity al . is (1985), it a receptor-mediated 2,3,7,8-TCDD, of et associated with many indicated by with receptor. the Some Ah the locus, toxicologic 4947 26 end points, however, shown in large genetic (Nagaya tissue et toxicity system. do n o t variation al. of do n o t cultures 1985, In 1986), addition, integration of Rozman indicates et the divided into the rats days the days in a p a r t i c u l a r is w e l l resulted have also of 2,3,7,8-TCDD. of normal rats, Male to the species although a complete of 2,3,7,8-TCDD has the normal days and to death also of demonstrated serum T4 in 2,3,7,8-TCDD was thyroid hormone modulation this in of Following and percent mortality 37 days and 80% , T4 survived receive lower mortality 2,3,7,8-TCDD normal the to b e The 2,3,7,8-TCDD 90 and of authors at treatment modulation energy metabolism. development and thyroidectomized in both The rats, to were the the levels rats and and had rats. study which ability therapy. death did not that a rats and considered by of the Sprague-Dawley to that days) T4 -supplemented the 89%, rats 63 of replacement similar between (35 results (T 4 ) h a s thyroidectomized received T4 that decrease mechanism described time thyroidectomized effects a monooxygenase Thus, the action thyroxine mean time a toxic 2,3,7,8-TCDD, It was in sensitivity. established, the this cells receptors species thyroid hormone rats of were (44%). these the of to mechanism with groups (mean of is i n d u c i b i l i ty levels respectively). Thyroidectomized longer susceptibility been there of the received T4 that that induction toxicity of administration 90 It h a s cells the a l . (1985) thyroidectomized at locus. characterized. that modulate the A h hydrocarbon hydroxylase and correspond this fully with lymphoblastoid parallels receptor-mediated mechanism not been human in a ryl 2,3,7,8-TCDD necessarily segregate with the lethal related participation toxicity needs to of further study. In addition, wasting syndrome investigated. such mechanisms and the Although end points, there only but to Further Target indicate the target higher In general, to lipid levels content (Ryan sensitivity of al. however, (1985), receptor of may be Further a there are Important not of in et also species determining and to that of needed toxic contain in r e gard target organ to determine of the species ability a mechanism has hot been to the Roberts this proven. et distribution this and issue. the toxicity. to m e t a b o l i z e sensitivity tissues. association 2,3,7,8-TCDD 2,3,7,8-TCDD-induced to 2,3,7,8-TCDD the clarify the to the than other. in p r o p ortion 2,3,7,8-TCDD. Is n e e d e d to nontarget of distribution to m e t a b o l i z e each few data organs of not end points, to are to the effects thyroid) other, for of necessary than effect the indicator that complete that than is relate There and rather toxic Further work capability certain responses liver observed processes distribute itself. suggesting these profiles. 1985b) the reported is the in the been been proposed result 2,3,7,8-TCDD al. to have as also understanding induces to such definitive organ (such as appears organ research data of a better species' sensitivity are the 2,3,7,8-TCDD between target of 2,3,7,8-TCDD a that 2,3,7,8-TCDD organs end points, cleft p a l a t e , have of mechanisms processes different of to b e organ/pharmacokinetic that relatively yet investigation understand how also h o w individual a number has underlining biochemical 2,3,7,8-TCDD. for development Although 2,3,7,8-TCDD chemical, the is data 27 Ongoing reported research. toxicokinetics and too the projects is 2.3.3.2 Monitoring Adequate TCDD There are a mechanism extensive for human analytical of biological methods materials concentrate 2,3,7,8-TCDD. Young available lipid content, 2,3,7,8-TCDD et al . used although for 2.3.3.3 The the development in h u m a n Environmental Analytical cannot"measure methods. are the on this indicate organic that in these the 2,3,7,8-TCDD potential years. of the Interactions that contaminants TCDD fate and gaps easily groups, Ongoing on analytical of most of other in various such Robens would exist and Zabik to is and (n.d.). Groups research organizations are available of to ability to in ambient air methodologies from 2,3,7,8-TCDD, are needed, strongly to of binding few data since soils At a a present, oral, gap existing of high to p e r m i t from dermal, Some progress media has there adequate or inhalation from conditions. and the the on The fate data regarding There are that its media. no data to other presence and and in recent environmental environment, conclusion the been made chemically with research develop to develop is the a of 2,3,7,8- the lack 2,3,7,8-TCDD of new of any will not focused primarily data. analytical small faster The methods amounts turnover research 2,3,7,8-TCDD Sweden, are monitoring of in the is in Centers Rutgers An for to and human Disease University, the example performed by in progress serum objective that will 2,3,7,8-TCDD time. investigation now being Substantial investigators in efforts more quantify very concentration of ability analytical co-contaminants media with research samples. 2,3,7,8-TCDD data interact Present environmental the is compounds. research determine methods observed results. in quantitative common support methodology an ongoing of (Patterson the of and volatility systems other identify the exceed bioavailability. doses environmental research. this unequivocally of with react with is low technology will the data transport. still in b o t h b i o l o g i c a l reactive with analytical of in envir o n m e n t a l 2,3,7,8-TCDD will under methodology serum present, to 2,3,7,8- 2,3,7,8-TCDD. b i o d e g r ^ d a b i l i t y , photolysis, indicate its absorbed 2,3,7,8-TCDD Substantial of two media. mechanism containing Environmental At existing can bind on transport the Bioavailability decreasing soils list detection resulting appears some thus of the thus tissues the sensitive and the or in human and-the bioavailability data to more investigate that reported samples. low content, estimation to in b l o o d significance insufficient exposure studies profile. lipophilic levels tissues Although on subject. that are Concentrations so levels media of considerations Bioavailability. environmental are of this reported method analysis, toxicological drinking water data (1984) recent environmental 2,3,7,8-TCDD interpret exists a number involved with 2 ,3,7,8-TCDD. The available 2,3,7,8-TCDD in m o n i t o r i n g b i o l o g i c a l monitor and detecting of in that in p a r t s - p e r - q u a d r i l l i o n 1987b). necessary be for large are samples are in biolog i c a l that action inclusion levels not relatively F e d e r a l. R e s e a r c h i n P r o g r e s s in milk Control, State j> 5oifO 28 University Division of New Yor k are involved at in Binghamton, such efforts and the (Ryan Canadian 1987). Food Research 3. 3.1 CHEMICAL The referred PHYSICAL INFORMATION IDENTITY chemical to CHEMICAL AND as identity 2,3,7,8-TCDD of 2,3,7,8-tetrachlorodibenzo-p-dioxin, throughout this document, is given to b e in Table 3.1. 3.2 PHYSICAL AND Selected in Table but physical 3.2. begins complete CHEMICAL PROPERTIES to and chemical 2,3,7,8-TCDD is decompose 5 0 0 ° C. within 21 at 800°C. degradation in presence a hydrogen-donating of 2 , 3 , 7-7U - T C D D the s at presence in organic properties stable of of t o ward heat, The decomposition 2,3,7,8-TCDD is ultraviolet light, solvents solvent. (EPA 2,3,7,8-TCDD acids, Gamma and are shown alkalies, is v i r t u a l l y susceptible to photo­ particularly radiation in the degrades 1984a). 4951 29 €> I 2.3.7.8- tetrachlorodibenzo[b,e]( 1,4)-dioxin; 2.3.7.8- tctrachlorodibcnzo-p-dioxin; dioxin; TCD BD ; 2.3.7.8- TC D D ; 2,3,7,8-tetrachlorodibenzodioxin; 2.3.7.8- tetrachlorodibenzo-l,4-dioxin (E P A 1985) T ra d e nam e N one (T h e com pound is not produced com m ercially.) (E P A 1985) C hem ical form ula c 12h 4c i 4o 2 W iswesser line notation T C 666 BO IO J EG FG LG M G or T C 666 BO IO J DG EG LG M G (H S D B 1987) C hem ical stru ctu re 0 o C hem ical nam e O Table 3.1. Chem ical identify o f 2,3,7,8-TCD D H H A t Identification Nos. C A S R egistry No. N IO S H R T E C S No. EPA H azardous W aste No. O H M -T A D S No. D O T /U N /N A /I M C O shipping No. S T C C No. H azardous Substances D ata Bank No. N ational C ancer Institute No. CP o\ 1746-01-6 (S A N S S 1987) H P 3500000 (S A N S S 1987) N ot assigned (H S D B 1987) 8300192 (S A N S S 1984) N ot assigned (H S D B 1987 N ot assigned (H S D B 1987) 4151 (H S D B 1987) N C I-C 0 3 7 I4 (S A N S S 1987) 31 Table 3.2. Physical properties of 2,3,7,8-TCDD Property Value References M olecular weight 321.97 Color Colorless EPA 1985a Physical state Solid at room tem perature EPA 1985a O dor Unknown M elting point 305'“C Schroy ct al. 1985 Boiling point 412.2°C (estim ated) Schroy et al. 1985 A utoignition tem perature NAa Solubility W ater (n g /L ) 7.91 (20-22°C ), 19.3 (2 2 °C ), 317 (2 5 °C ) Schroy et al. 1985. Adam s and Blaine 1986, M arple et al. 1986a o-Dichlorobenzenc, 1400; chlorobenzene, 720; benzene, 570; chloroform , 370; m ethanol, 10; acetone, 110 Schroy et al. 1985 ■Density (g /m L ) 1.827 (estim ated) Schroy et al. 1985 P artition coefficients Log KoW: 6.15-7.28 Log K ^: 6.0-7.39 EPA 1985a, Schroy et al. 1985, Jackson et al. 1986, M arple et al. 1986b V apor pressure (m m H g) 3.46 X lO'9 (30.1oC ), 3.51 X 10'9 (30.2°C ), 1.4 X 10'9 (estim ated at 2 5 °C ) R ondorf 1986, Schroy et al. 1985, Palansky et al. 1986 H en ry ’s Law constant 2.1 X 10'6 atm -m 3/m o l (estim ated) Schroy et al. 1985 Flash point Unknown R efractive index Unknown Flam m ability limits Unknown O rganic solvents (m g /L ) Conversion factors Vapor Liquid Solid 1 ppb = 13.384 /tg /m 3 at 20°C 1 ppb (w /v ) = 1 fig/L ; 1 ppt (w /v ) = 1 n g /L 1 ppb (w /w ) = 1 #ig/kg = 1 n g /g ; 1 ppt (w /w ) = 1 n g / k g = 1 P g / g ‘'N ot available. 4953 //[/] f a - y » 4. 4.1 TOXICOLOGICAL DATA OVERVIEW Human and animal data following ingestion, and to the of soils, addition, extent animal data through the tissues in p r o p o r t i o n skin. placenta with exposed excretion of in and the through lactation. and from 11 TCDD toxicity, toxicity of year the >1 and have from acute /ig/kg f o r for a A dose for however, no The toxic this may In wasting prior to pigs newborn can be lactation. result primarily in also in which to the 2,3,7,8-TCDD feces is as w e l l differences resistant is as in b o t h varying to sensitive oral to other in (total) essentially in species experiments to study from the only have been to been 2,3,7,8- to the is dermal end The point. are dermal major toxic effect death, is with no clear observed in signs oral acute 275 are acute indicated L D 50 dermal to b e s tudy after of monkeys rabbits /xg/kg; only toxicity of produced This studies. in humans lesion Although the strains animals all 90-day observed oral produced and after the extremes. limitations lesions in humans, ears in w h i c h Subchronic The to b e effect. on the two a other conducted. chloracne. there from with number in contaminant reported 2,3,7,8-TCDD vary those and a no respectively, reported tested, demonstrated a as as been during 2,3,7,8-TCDD exposure solely these small specific syndrome, of in death. not been clearly lesions a to monkeys, animals, syndrome of chloracne, the doses same rabbits has have 2 , 3 , 7 , 8 -TCDD and not have exposure the study sensitivity effect 2 , 3 , 7 , 8 -TCDD there and hamsters, 3 /ig/kg r e s u l t e d 2 limited (observed this were toxicological be Lethal of effect different cross elimination half-lives e x p o s e d to feeding only available the can through relatively monkey, 9-month intermediate following is distributed during to In the m e t a b o l i t e s strain L D 50 v a l u e s b e t w e e n 2,3,7,8-TCDD Inhalation and chemicals, guinea cumulative guinea pigs that exposure. tested having L D 50 v a l u e s been industrial 5000 value in of elimination with which deaths exposure. rates, and thought removal species hamster, 0.6 with are fetus, Unmetabolized intestinal There the to currently is 2,3,7,8-TCDD 2,3,7,8-TCDD rapid and urine. direct Although humans species is the -50%. absorbed well 2r, 3 , 7 , 8 - T C D D content. of is absorbed affected by binding decreased by 2,3,7,8-TCDD of is can be 2 , 3 , 7 , 8-TCDD. of herbicides to exposure elimination in lipid relatively bile 2,3,7,8-TCDD only absorption, the 2,3,7,8-TCDD through days to is that redistribution excreted metabolism indicate subsequent detoxification that although bioavailability absorption Following following Metabolism indicate is in models in a n i m a l s expression of mice, are investigating of and are this lesion rabbits application). of 2 , 3 , 7 , 8 -TCDD progressively altered species organ tested, it exposure lose body function. occurs is the weight Although predominantly 4955 33 a systemic animal 34 only at not been doses that observed are lethal Immunotoxicity has species and may be Immunotoxicity, exposure near one also b e e n of however, Hepatotoxicity has there is The human data are variations two m o s t sensitive severe Other observed in not evidence that completely in hepatic organ lesions systems are extensively are additional as those prime indicators of into extrapolating Many - ' o f t h e extensive also differences may on has animal 2,3,7,8-TCDD. to 2 , 3 , 7 , 8 -TCDD effects be related for to a into the Once in complex. protein is strain and known translation genetically differences segregation. The in in of the it of DNA may be toxicity protein levels differ. These explain of species differences 2.3.7.8- TCDD toxicity, receptor while The evidence herbicides explain all and others from other 2.3.7.8- TCDD is toxicant. induces a variety The data industrial determine additional in of male a scheme of which receptorthat The both receptor : locus, and many of explained by widely dispersed although differences may help The receptor strain differences in responses segregate with to chemicals known demonstrate that data from studies in the these adversely clearly is a human laboratory mice that and 2,3,7,8-TCDD 2,3,7,8-TCDD affects causes to h e r b i c i d e s are reproductive that and not male to in is abortions. hormone other adequate health 2,3,7,8-TCDD spontaneous 2,3,7,8-TCDD' affects reproductive to contaminated with 2,3,7,8-TCDD in species exposed demonstrate and exposed to b e abnormalities. populations species that and have of the protein, sensitivity. toxic contaminated with studies several to b e to These are species, of human populations demonstrate 2,3,7,8-TCDD evidence function The from human Animal fetotoxic studies of developmental chemicals if as insight do not. inadequate clearly species the as to be observed proposed the A h quantitative some industrial developmental however, because and in man. follows in m i c e in different the been is is special those cell with in the affected; appear any receptor is k n o w n quantitative cannot not affected. observed systems mechanism to There be system sensitivity. nucleus, and do mechanism segregate that relatively investigated provide cytoplasmic organ the they or in This in different some digestive animals nucleus receptor addition, indicate system may not been the to In in humans. exposure receptor-mediated mechanism soluble 2, 3 , 7 , 8 - T C D D mixed 2 , 3 , 7 , 8 -TCDD. discussed have the subsequently migrates transcription Also, toxicity differences to since species. the later. 2,3,7,8-TCDD. 2.3.7.8- TCDD binding on have end points strain species, occurs effects nervous systems observed animal and m o n k e y s , develop affected by the of also occurred. other effects organ described and -toxicity proposed however, pigs that 2,3,7,8-TCDD toxic species effect chemicals than many data These clear, species, guinea there the syndrome of of related a variety this induction of hepatotoxic in animals, kidney. humans. a variety been in h u m a n s rats, wasting effects directly evidence Ah in sensitive not been suggestive this observed the m o s t other potentially hepatotoxic the The has suggestive species less lethal. in h u m a n s . and to or in h u m a n s . There levels is and organs. f ) \ ■:*- '¿'“-.v 35 Although results in bioassay agent; 2,3,7,8-TCDD has genotoxicity systems however, the predictive of genotoxic agent. The cohorts be alone wastes, to but which 2,3,7,8-TCDD, recognized these of 2,3,7,8-TCDD positive evidence and and that provide As 2,3,7,8-TCDD to materials in as 2,3,7,8-TCDD as as well the p ublic and also gaps the this dibenzofurans problem, Technical and recommended from exposure gaps are an to filled. Panel of the interim method to only carcinogen. The is The data of a whereas lower toxic potency values were developed using congeners and of most Using this of TEF. toxicity reproductive and determined by weight of the w e i g h t the of of placed used, most used, not of of types similar the that carcinogenicity the for tests evaluating a mixture of extrapolation available, e n z y m e s ) . The additivity as the relative resulting in vitro oxidative data the followed by tests defined evaluation data available, from from is appropriate levels are v e r y of an reflecting the h u m a n evidence toxicity of on the are assuming congeners into for data exposure to extrapolate between data on data a means are These the above induction of relative to data were effects the congeners. segregated data w o u l d be carcinogenic particular can be human are and thus to data equivalence results 2,3,7,8-TCDD less, used definitive this for or risk the dibenzo-p-dioxins the values of.reliability (estimated if none generated and were and developed the used until "2,3,7,8-TCDD approach were Forum has dibenzo-p-dioxin TEF unity tiered degrees If carcinogenicity estimating a system, reproductive and The In estimating can be relative 2,3,7,8-TCDD would be the Finally, data, 2,3,7,8-TCDD toxicity data differing congener are in chlorinated chlorinated TEFs 2,3,7,8-TCDD would provide establishing other The are 2,3,7,8-TCDD. unity, data. all for (EPA 1987). to dibenzo-p-dioxins/Chlorinated the toxicity information with which exposure generates 4.1. existing exposure procedure in Table level from This of of resulting that isomer-specific congeners concerns mixtures TEFs other 2,3,7,8- industrial EPA has chemical complex mixtures to ash or The these of is n e v e r fly assisting and distribution data 2,3,7,8-TCDD Risk Assessment for (TEFs) interactions a human in known materials. from congener- TCDD). chemicals toxic Chlorinated factors" animal is that available based a is reported the h u m a n environment in potential of findings incinerator toxicological the to process. a carcinogen along with many the hu m a n health equivalent a result, other, p o t e n t i a l l y response relating usually is n o t evidence the such contain compounds, presented diverse genotoxic assays 2,3,7,8-TCDD clear in a in is results chlorinated a is 2,3,7,8-TCDD negative other evaluate on that positive results 2,3,7,8-TCDD negative suggest produced positive in animals. Exposure TCDD that are that 2,3,7,8-TCDD. however, carcinogenic numbers to h e r b i c i d e s suggestive data, consistently there suggest agents evidence contaminated with animal to large with both exposed provide may genotoxic human conflicting, which not assays, data on in 2,3,7,8- the TEFs TEFs toxic are (with receptor containing for however, binding thus response, for a known 2,3,7,8-TCDD. 4957 36 Table 4.1. Recommended T E Fs° for 2,3,7,8-TCDD and its congeners E PA current recom m ended values based on various effects Compound M onoCDDs 0 DiCDDs 0 TriC D D s 0 2,3,7,8-TCD D O ther TC D D s 1 0.01 2,3,7,8-PeCD D s4 O ther PeC D D s 0.5 0.005 2,3,7,8-HxCDDs* O ther H xC D D s 0.04 0.0004 2,3,7,8-HpCDDs* O ther H pC D D s 0.001 0.00001 O CD D 0 “Toxicity equivalence factors. 6Any isom er th a t contains chlorine in the 2,3,7,8 positions; CDDs = chlorinated dibenzo-p-dioxins. Source: E P A 1987. 37 4.2 TOXICOKINETICS 4.2.1 Absorption 4.2.1.1 Inhalation Pertinent or animals data regarding following the inhalation absorption of exposure were 2,3,7,8-TCDD by humans not found in the available literature. 4.2.1.2 Oral Human. of The c o m oil at a of of of the Animal. pigs, in a rates with absorption at h i g h e r the 1975). The vehicle on Poiger and its solution in et 1980). repeated dosing, dose levels. feces study in 1986). and urine, as w e l l samples, indicated absorption, the h a l f - l i f e days. (1985a) when elimination ^ H - 2 ,3,7,8-TCDD Schlatter the 2120 al . show that administered by gavage There but rats, (Piper there was 50 2,3,7,8-TCDD to appeared Absorption of also between McConnell et al . 2,3,7,8-TCDD of et to b e some is guinea al. no 1973, change decrease in in 2,3,7,8-TCDD when and 60% (Fries (1984) from c o m TCDD-contaminated TCDD, from (Phillipi et the Umbreit al. is et less al . et the al. toxic indicate the and Marrow soil and of compared with a activated carbon. Since to soil, (1986) compared the absorption that from 2,3,7,8-TCDD effects an showed an substantial type -50% that equivalent soil h a d and time b e t ween soils the less from 2,3,7,8- amount of on absorption of occurs contact, and may 2,3,7,8-TCDD substantially obtained from different sites. Dermal Human. Pertinent dermal data exposure regarding in h u m a n s the were absorption not found literature. ■ ' as extraction soil 1982), and influence duration decreased from the adsorbed Phillipi 2,3,7,8-TCDD (1985a), to b e than that EPA occurred when (1986a) to substantial in absorption absorption was demonstrated Huetter of of soil with increasing contact 1981, in suspension likely that b i n d i n g that described absorption liver, however, data As indicated by biological the data soil; absorption contaminated following oil. These the efficiency with As in soil was contaminated 4.2.1.3 oil. soil suspension and Lucier suggesting bioavailability. no contaminated in c o m from suggested whereas a administered has a decrease in a environment 2,3,7,8-TCDD than 2.3.7.8- administered the is absorption. observed a d m i n i s t e r e d as in administered 2,3,7,8-TCDD (1980) ethanol, TCDD affect in lipophilic vehicle in w h i c h when 2.3.7.8- from 80%) gastrointestinal 50% compound was soil and Following be from an fat v i a b i o p s y in E P A to diet was Schlatter 2.3.7.8- TCDD amount are ingested • effects of in absorbed. (5 0 Olson absorption in (Poiger elimination reviewed absorbed 1979, administered ng/kg calculated.to Studies al . available sequestering dose w as or hamsters Nolan_et 1.14 of elimination was generally well data a male volunteer cumulative determination that >87% for in which a dose Measurements as absorption 2,3,7,8-TCDD in of 2,3,7,8-TCDD the 4959 available 38 Animal. Data 2,3,7,8-TCDD absorption "i n E P A rats is regarding absorbed the v e h i c l e can ( 1 9 8 5 a ) , it w a s when vaseline the estimated or p o l yethylene respectively (Poiger or in soil observed studies, with 40% of in 1980). animals although affect resulted carbon in skin, the dose was whereas 1.4 and absorption As oral discussed absorbed by in absorption, 2,3,7,8-TCDD in poorer of <2% limited. with application 9.3% Applying resulted are as absorption. in methanol, Schlatter activated in o r a l the that applied glycol and absorption through substantially compound was paste dermal well as absorption a than and n o n d e tectable, respectively. 4.2.2 Distribution 4.2.2.1 Inhalation data regarding the distribution of inhalation exposure Pertinent of humans and animals were not (1986) estimated available 4.2.2.2 Oral of ;- Poiger and Schlatter 2,3,7,8-TCDD was sequestered i n g e s t e d ~ 3H - 2 , 3 , 7 , 8 - T C D D in study, corn lasted 135 days, only during the initial -data would be consistent with 2.3.7.8- TCDD in humans, as Adipose 2.3.7.8- TCDD tissue in this of Vietnam veterans 10 the (Schecter Weerasinghe group with is al . et al. subjects with no on order to fat kidney below lipid basis of (-6 the data were subjects cannot be Animal. described pigs. In the rats to and (1985a) mice, In et since two h u m a n s to only fatty the entire reviewed the of two from the were studies of up to study b y - : correlation adipose autopsy. levels data tissue observed further levels a in examined They in descending in muscle compared and on a per should be examined and syndrome; therefore, one the population. of in however, congenic tissues to h a v e distributed liver contained observed were subjects were a number fat; patterns at These 2,3,7,8-TCDD liver Adipose ■ subjects that (1985b) 2,3,7,8-TCDD tissues. of occupationally a clear levels al. levels another apparent 2,3,7,8-TCDD followed by Similar administration. Ryan of from daily difference between Although it was in if control any The and pharmacokinetic reported detect detected potential exposure. however, body During (1986) Orange^and humans however, distribution ng/kg. al. 1984); the treatment. determine detectable that suffering et tissue, to (-2 p p t ) , w i t h caution, EPA Geyer subjects. similar between was following liver generalized 2 . 3 . 7 . 8 - TCDD, reversed. failed exposure. detection; 2 days in unexposed demonstrated, and 1.14 2,3,7,8-TCDD were 2,3,7,8-TCDD ppt) of of a volunteer radioactivity was adipose e t al. a weight basis interpreted with of of Gross known dose correlated with 2 , 3 , 7 , 8 - T C D D , :a n d distribution determined not a at to A g e n t 2,3,7,8-TCDD (1986) after examined and unexposed was for been sources of fat the h i g h b i o c o n c e n t r a t i o n reservoir 1985, of veterans exposure a depot the level et the elevated in hu m a n has exposed to p o t e n t i a l times following the that -90% in calculated by assumptionslevels exposed oil which in'the blood intake in .. this models. found literature. Human. burden 2,3,7,8-TCDD studies mice, the greatest in guinea after mice early rats, that and guinea amount pigs this of was intraperitoneal which differ only at the Ah 39 locus, the distribution patterns described, except reported the in nonresponsive appears that that livers strain there of little distribution pattern of that the tissue transport in accumulation in lymph the lymph the fat study, -0.5% of the first crosses dose was (1986) fetuses, Nau et the of the y o u n g of Again, fetal al. 2,3,7,8-TCDD fetal fetus. and in 1 .6 was the mouse 1985). In this results were 2,3,7,8-TCDD by not uniform, In addition (1986) reported after to p r e g n a n t m i c e liver was of for liver Similar the milk and pup . levels The h a l f -life and Birnbaum liver. reported through high it the (1986) and accumulates the Thus, in progressive tissue treated with occurred via the 2 4 h. in 1986). al . initial distribution was the of and!17. in m i c e in exposure with (Weber detected Fetal et were the difference is m e d i a t e d adipose placenta located exposure 14 the administration Birnbaum strain rapidly with of injection. administration rat, the the 2,3,7,8-TCDD the previously 2,3,7,8-TCDD compared with Lakshmanan out reported by Krowke subcutaneous for during gavage mice 2,3,7,8-TCDD in those of 1 9 8 3 a , b; 2,3,7,8-TCDD respectively. after al. decreasing and 2,3,7,8-TCDD of system of fetus et to levels responsive 2,3,7,8-TCDD. redistribution 5.3 weeks, similar higher potential distribution in the 2,3,7,8-TCDD the (Gasiewicz is were relatively with to 75% of in utero that postnatal a single between gestation the p r e d o m i n a n t days storage site. 4.2.2.3 Dermal Human. compounds Sect. 4.2.2.2 available are Studies known not (oral human clearly although oral 4.2.3 for easier data. Routes of likely route inhalation studies exposure are are other were exposure in likely the these is p r o b a b l y also industrial discussed comparison with most the or 2,3,7,8-TCDD to in only studies dermal, occur. available. ------- Metabolism inhalation exposure or data are regarding dermal of humans, regarding animals exposure were formed they glucuronide and metabolism of humans found the m e t a b o l i s m is are of in or the of slow excreted Recently, however, 1985a). of the radioactivity 2,3,7,8-TCDD relatively rapidly conjugates EPA was animals, or following following literature. following in the to m e t a b o l i z e , oral However, oral exposure of once metabolites the urine and bile as (Olson et al. 1980, 1983, as reviewed reported that up to Olson tissues used, the rapidly previously metabolites believed. of of (1986) guinea pigs 2,3,7,8-TCDD were Sawahata al. h y d r o x y - 2 ,3,7,8-tetrachlorodibenzo-p-dioxin and in vitro, and Poiger 2-hydroxy-l,3,7,8-tetrachlorodibenzo-p-dioxin of 2,3,7,8-TCDD. Other treated with indicating et trichlorodibenzo-p-dioxin but through associated with metabolites, conditions metabolites 2,3,7,8-TCDD available 2,3,7,8-TCDD sulfate in as not the available. 2,3,7,8-TCDD are distribution) to h e r b i c i d e s with experimental No Pertinent data exposed contaminated defined; and Animal. of humans to b e (1982) labeled under eliminated identified the as 1- 8 -hydroxy-2,3,7et in hydroxylated not that 2 8% al . dogs (1982) as products identified major were ident | | j | { l 40 as minor metabolites. metabolites activity and than al. (1982) bile of Mason 2,3,7,8-TCDD, which supports the Poiger et al. guin e a pigs, respectively, and for the (1983b) reported urinary responsive increase, large 1985, respectively, and Wroblewski differences in of metabolism metabolites - the available 4.2.4.1 data regarding exposure literature. However, Poiger oral and background levels 3 days 7 to 125, 2120 It is, actually parameters 4.2.4.2 during only of These data and and and rats 320% subsequent dose pretreatment with of a subsequent Wroblewski suggest and Olson that some of the related to the rate quantitative excretion data or of differences and 2,3,7,8-TCDD animals regarding of humans the were in the following not found excretion of animals are would studied 3.5% days in the 2,3,7,8- available. change elimination single 135 days the individual with 11% of radioactivity were the was dose dose was the b o d y w a s eliminated. calculated not known with any studied. eliminated From and of above Fecal in the during these assuming certainty first-order kinetics, of dose never nonabsorbed material), whereas the the profile 1.14-ng/kg rapid, of in the ingested a of to b e eliminated by and appear to b e by oral the the levels the however, 2,3,7,8-TCDD in no or observed and the data, a first-order that 2 , 3 , 7 , 8 -TCDD other kinetic estimated half-life. metabolites apparent differences whether the bile, derived Olson elimination (1985a) of are in a n intraperitoneal in probably was EPA its urine major demonstrated by mice. of a 100 of for (in v i v o ) of metabolism 1985, al. Animal feces that rate et similar in a (in v i t r o ) , of humans Urinary (speculated kinetics. feces In dogs of metabolism could Gasiewicz elution profiles resulted 2,3,7,8-TCDD may be (1986) initially half-life were 2,3,7,8-TCDD 2,3,7,8-TCDD were Schlatter a volunteer who chemical. elimination was first exposure Schlatter from tritiated the of in m etabolism the the and Human 2,3,7,8-TCDD is of rats 2,3,7,8-TCDD. toxicity. of mice. qualitative dermal or following days of to et from formed. Pertinent the of these Excretion -inhalation TCDD as the and 1985). toxicity as w e l l the metabo l i s m 2,3,7,8-TCDD on (Poiger of differences in guin e a pigs effect amounts evaluations rate toxic in strains in the whereas 2,3,7,8-TCDD Olson 4.2.4 qualitative less in of biological extracted if difference some less observation by Weber metabolites equivalent determine pretreatment with 2,3,7,8-TCDD had no of to and nonresponsive 2 , 3 , 7 , 8-TCDD, dose than observed that that 2,3,7,8-TCDD were and biliary metabolites (in vitro), of (1982) strain differences investigated account the synthezised considerably and Species (1986a,b) they had treated with been Safe that dogs have and demonstrated routes (1986) in 2,3,7,8-TCDD reported first-order process. the material (EPA 1985a). unmetabolized from eliminated predominantly direct g u inea pigs. through that b e t w e e n Nau 91 and 99% metabolites detected in elimination, as et lactation administered Although 2,3 , 7 , 8 -TCDD intestinal was al. in There (1986) the reported is significant of the excreted in 41 2.3.7.8- TCDD-derived guinea fecal in pigs; feces and days); 15 in radioactivity was 72% was the strain ( 17 mice days). follows to 31 24 (1986) there there that are 4.3 indications with a possible Lethality 4.3.1.1 and Pertinent data inhalation not in found 4 . 3 . 1 .2 the No Animal. mice of mice, with greater was observed strain; are no (1 1 to 59% also order): 24 11 excretion is v e r y of >1 year and varied with days (2 2 to in and hamsters longer half-lives 2,3,7,8-TCDD rats guinea pigs days, strains); adequate half-life and elimination in nonresponsive of >90 data (10 days in from m o n k e y s , persistent in this (EPA 1985a). lipophilic Hartley and of humans or available in guinea McConnell toxicity dose longevity to 2 , 3 , 7 , 8 - T C D D were' pigs 1985). 70 Following acute of et that some after gavage 1973), from 20 5000 pg/kg also been rhesus Eight 1985); at a time of toxic total of ranges 60 pg/kg pg/kg 16 as independent 5 to of but in in in rats, (EPA 100 1985a, the lowest Reproductive— are very a sensitive monkeys died 9 -day period indicated by of 0.2 observed 40 dose levels orders pg/kg pregnant dose generally from 2.1 1978). over effects, is to monkeys 4 administration 0.6 al . lethality low cover studied, et 2,3,7,8-TCDD death that extent, 1.0 to acute in hamsters (McConnell dose the at very lethality range have of toxic in female administration, to is 2,3,7,8-TCDD. (McNulty of to al . to lethal total observed to be, L D 50 v a l u e s , monkeys was a differences 1000 indicate extended period appears and Rhesus gestation this reported (Schwetz treatment with were chemical level. effects abortion, investigations that Species pg/kg, studies lethal available. extensive were in mice, tested, during decreased animals literature. are of tested. solvents, 600 p g / k g after lethality exposure indicate species the Longevity regarding Results of magnitude dose Decreased studies 2,3,7,8-TCDD in all an feces of Oral Human. to the Inhalation following to for in feces TOXICITY 4.3.1 of C57B1/6J reported even guinea p i g s . Although species, the (in d e c r e a s i n g days); days found in Half-lives as and Olson detected responsive of hamsters. rats responsive to and excretion the species 30 54 days; after pg/kg. only this a after lag time threshold is reached. Other of factors 2,3,7,8-TCDD. with responsive besides These C57BL/6J 2.3.7.8- TCDD-induced mice (Gasiewicz et al. 1983a,b), pg/kg) reported (1985). but 320 Strain would affects be the differences anticipated acute bioavailability of to lethality the as and for the DBA/2J to b e strains not been occur. Also, However, the of in lethal of rats the as acute animal twice as toxicity tested, sensitive to strains dose (ranging (Walden investigated 2,3,7,8-TCDD, compound. affect strain nonresponsive differences four have of differences include demonstrated lethality 165 to species variables in and other species vehicle substantially probably by discussed by from Schiller altering Kaminsky the et 42 al. (1985) influence the and illustrated by Umbreit of m a trix properties matrix this of the m a t r i x substantially issue is administered suggesting the charcoal some of (1985, 1986a,b), complex, duration with The the p r o t e c t i v e the lethality 2,3,7,8-TCDD the effects are of subchronic of of orally subcutaneously result al. as the complexity (Manara et the such with effect of the factors of contact bioavailability. on administered al. is as illustrated by activated that as w e l l altering further intraperitoneally el on bioavailability or 1984), of processes other than bioavailability. The lethal studied by were maintained TCDD for 90 to At were consumption that observed of male guinea et al . on diets days. respectively, total effects DeCaprio 76 in no in 5 of contained 500 of it. — deaths. to No studies There 2,3,7,8-TCDD by the 531 acute /xg/kg) 12 is the LD50 of in N e w administration, between 4.3.2 and the 22 one dermal of the is acute Allen et 9 months the 0.0093 to 2,3,7,8-TCDD to available in exposure al . on (1977) a diet total 0.014 death was regarding et acetone rabbits. As al. that dose ^g/kg/day. Toxicity the (1973) 275 was observed /xg/kg with lethality determined was protracted, Organ (range, after deaths 142 oral observed Chloracne Pertinent the available in Oral, human. Animal. or No Greig administration severely acneform of This (1984) 2,3,7,8-TCDD in than 4 rhesus was diet at also produced lesions 1977). This dose not since the were subcutaneous monkeys that development exposure and and lesions single The of were not given a toxic, single exposure to of dose facial of to b e observed 2,3,7,8-TCDD gross of monkeys to /xgAg/day) for (Allen a l. formation of other hair. and in m a n y 0.014 associated with loss (1978) chloracne the appeared al. resulting (0.0094 for et in male gavage females McConnell resembled threshold severe edema ppt a /xg/kg. Longer-term 500 months a 75 skin after males. severely death. typical weeks at the in was the inhalation available. observed mice dose the lesions are 2,3,7,8-TCDD and ultimately regarding following literature. studies affected eruptions /xg/kg. studies animals hairless A2G-hr/+ as the similar intake, Schwetz Systemic/Target found such study route. treatment. humans effects 2,3,7,8- 70%, available. only time by 70 are after Inhalation. at of and o f 0..61 n g / k g / d a y for food ppt 10 been pigs 0 . 8 /ig/kg f o r however, 3 /xg/kg o r Zealand white chloracne more From Guinea This value study maintained to of Exposures days 4.3.2.1 female 2 of calculated. described pigs. or 430 LD50 value 2,3,7,8-TCDD. to b e 76, monkeys, 8 animals 2,3,7,8-TCDD have Dermal Human. that In rhesus calculated Animal. of an 10, to guinea mortalities 2,3,7,8-TCDD. deaths 4.3.1.3 and the p r e v i o u s l y to observed ppt 2, ppt, 2,3,7,8-TCDD was in pigs exposure in H a r t l e y containing and 430 observed, of resulted ingested was (1986) In et 9 chloracne, dermal effects addition, this ( *0 iP AQ 43 level of 2,3,7,8-TCDD Dermal, observed to human. a variety of has TCDD known is after to b e define doses the chloracne area. have to been rather related than to a Additional small groups fatigability, et (on 1975). of which in use, the Dermal, the mice developed resemble Toth et for al. was in a only the to al . (1982) to this would be General that chloracne, mice skin glands, exposed certainty that chemicals cases, is component that compound such as w i t h "inert ingredients" applied 3 0.1 of pg times/week for 2,3,7,84 w e e k s , — T h e ----- examination to and buildup with the of next most inducing a dose in this of keratin agent the into seven effective response at 200 study precludes ¡ig the 0, 0.7, kidney, groups. although is 2,3,7,8-TCDD 0.007, 7.0 spleen, and The it similar to or dermatitis is to not clear Swiss mice by pg/kg/week. liver, had if chloracne, along with some the etiology then the is LOAEL in pg/kg/week. Chloracne induced by Chloracne to 2,3,7,8-TCDD a minor chlorinated in many Schulz observed with other other effective response, single anger 1957, included hyperkeratinization, the most the dermatitis discussion. can be in humans. of in the of easy sleep dose. treatment 0.007 disorders, appeared on histologic the administered doses observed all t h e .same. If of a as The of include product. Skh:HR-l dermal to only state with but, as w e l l final et threshold at exposure include contaminant, sebaceous use commonly 2,3,7,8-TCDD. studies signs neuropathy, and are the difficult not this may during play in case digestive Kimmig symptoms solvents of (1979) similarities TCDD is Puhvel The 1 year dermatitis, humans is this of a Amyloidosis lesion loss, 3,4,3',4'-tetrachlorobiphenyl, application. soils These headache, 1961, since from of hairless inducing reported 2,3,7,8-TCDD being 2,3,7,8-TCDD was determination mice and with but the young. 2,3,7,8-TCDD. these in in gavage of which compound, children Schulz Changes tested accident, adults, bouts exposure absences persistent, al . et lesions cysts. In weight skin dermal than and upper can be to - c o n t a m i n a t e d libido, chloracne. hyperplasia, and to that to face and uncharacteristic petroleum skin the changes, produced animal. to related on Seveso available is k n o w n Bauer formulation of TCDD to of b a s i s ) , it 2,3,7,8-TCDD herbicide loss chemicals, concomitant the these 2,3,7,8- in producing not effects frequently been exposure of (1985). It disfiguring After appetite, Many are very are appear toxicity have been exposure, symptoms provide per to a mixed a percentage these used 1971, exposure always usually exposure exposed of sensory al. 1957 ^'Oliver __ a c u t e loss prevalence lesion. other visible lesions has accidental Suskind exposure this sensitivity of insomnia, disturbance, any greater signs The compounds on tested. chloracne potent data 8 animals of the m o s t exposure. greater time and chloracne more of people aching muscles, (Poland to century, the compounds. to p r o d u c e The the from 5 of (1979) after to of of Taylor can be develop deaths aromatic lesions lasting many years appeared of prior exposure. These turn sufficient necessary appears 2,3,7,8-TCDD one the few days reviewed by however, in the a chlorinated been chloracne; trunk Since in h u m a n s lesions resulted is chemicals single the is a persistent or multiple first toxic contaminated with deformative exposure effect usually 2,3,7,8-TCDD ^ to skin 2,3,7,8observed and 4965 in appears . 44 to b e a sensitive can develop but data are chloracne toxicological chloracne not available Hoses and Prioleau and incapable of providing a compounds lesion and studies by suggested animal little As in appetite the to pigs to et are al. in typical of this In vitro epidermal dermal syndrome lesions in humans not found and Potter from or in that total al. the studies of in food loss final cold-adapted intake syndrome, as fast as treated rats monkeys exposure after decreased diets 1986). Weight 2,3,7,8-TCDD at -0.01 pg/kg/day months. eight monkeys (10 m a l e s provided doses in b ody a group and of weight, loss (the females) whereas 40% decrease died or were only and in dose guinea pigs in b o d y weight. killed which maintain support accounts lost weight lose et monkeys level studied) died at this on diets All doses for 90 a before 9 days 15% 0.026 guinea pigs (when m o r i bund) 1977, Ingesting for of after al. dose. pg/kg/day had given twice 1986). body weight (Allen female five 0.0049 mechanisms death. (Rozman and Greim maintained of other days accounts partially rats also -20% tested 2,3,7,8-TCDD only 2,3,7,8-TCDD was few 2,3,7,8-TCDD, cold-adapted temperatures Treated a ultimate rats, Lu only until that and the were 10 but al. however, consumption Sprague-Dawley guinea pigs rats energy et of weight. food in w e i g h t containing al . loss treatment with at n o rmal food Seefeld body weight consumption since altered 1984; food and Hartley to or nutrition from until pair-fed parenteral decreased decrease with Recently, stable after pattern, 1986). partially species (EPA 1985a). 2,3,7,8-TCDD-induced Peterson et several decrease Studies than malabsorption Seefeld in 2,3,7,8-TCDD a biphasic in b o d y w e i g h t for /ig/kg/day h a d of cultures for observed of observed pigs epidermal body weight will possibly relatively wasting high-dose been doses results guinea pigs the decrease has lethal syndrome 1985; for Guinea to available. exposure. demonstrated the Only limitation 2,3,7,8-TCDD were after rather that which to of et 2,3,7,8-TCDD. exposure relationships. the that Rhesus from s k i n was understanding receptors in the to upon regarding wasting exposure review, 1984; observation prolonged the results syndrome days the DeCaprio route, induce recovered exposure the using human demonstrating levels had chloracne death, Additional high 30 maintained account who in a poor affect data acute above (Rozman Kellin for most must to examination of dose-response 2,3,7,8-TCDD, protected Hartley prior of the w a s t i n g (1986) guinea humans any exposure. studies wasting suppression 1984a,b; on ultimately No of 15 that utilization al . necessary in humans; resulted (1984) inhalation The occurs al. Pertinent human. described suggest that literature. administration et et administration death believed syndrome following Animal. after dose of past develop lesion has 2,3,7,8-TCDD Inhalation. Oral, this information which Vastlng available indication 2,3,7,8-TCDD may associated with the the is 2,3,7,8-TCDD by studied humans animals models Greenlee that factors, animals It to that histologic any that produce appropriate 4.3.2.2 regarding (1985) determined few experimental growth exposure in h u m a n s . chloracne Only e nd point. following the in the end of 45 the study. that even The in 0 . 0 0 0 6 1 -/ig/k g / d a y long-term characteristic levels that Dermal. animals of are studies, exposure lethal to Pertinent exposed to level the was loss 2 , 3 , 7 , 8 - T C D D , is the level. It appears of body weight, associated with dose animal. data regarding to 2,3,7,8-TCDD dermally a no-effect severe the wasting were syndrome not found in h u m a n s in the or available literature. General of discussion. 2,3,7,8-TCDD observed wasting syndrome in e x p e r i m e n t a l acute longer-term in b o t h associated with The toxicity and lethal doses. The is a characteristic animals. studies wasting This and syndrome is m o s t syndrome has sign is commonly not been observed in h u m a n s . Since the mechanism determined whether TCDD toxicity effect that is the can be proposed by Aust the thyroid which has an the-mechanism be afT i n d i c a t o r toxicity. No (1984) of in are No The toxic acute (EPA porphyrin. These exposure and number 2.5, necrosis, or an been 2,3,7,8-TCDD on 2 , 3 , 7 , 8-TCDD- weight. an early as Regardless syndrome appears sign a result to of of 2,3,7,8- not in humans found in or the of liver, Lesions or have (1983), severity or incidence was fi g / k g in males Hartley in maintained were (DeCaprio not et on diets observed al. 2 ,3,7,8-TCDD consist of observed of 5 to following observe of morphologic and 4 to 6 Deaths reported were the next Although and reported liver doses lower chronic In a of at 0.1, guinea pigs, increased for doses 9 0 -day changes dose changes steatosis, in all dose respectively. that provided a females, 2,3,7,8-TCDD including hypertrophy, were the 2 0 0 /jg/kg> 1 male dose in guinea of necrosis, generally been similar mild 1986). to duct bodies at and mide amounts association between guinea pigs, rats changes excess and gavage females, to severe bile of reported. and of rodents did single no alterations, (groups Changes an 2,3,7,8-TCDD however, /jg/kg- and hyalin-like study with in doses given a 20 of whereas membrane guinea pigs and changes available. administration effects of Mg/kg/day than change lipid metabolism, indication animals the acute Collins reported) 12.5, in no liver of body the w a s t i n g rather at r e l a t i v e l y h i g h small 12.5 of conjunction with 2,3,7,8-TCDD were are cellular liver Turner strain not focal effect animal have regarding hepatotoxicity to 1985a). altered although the in loss administration observed upon proliferation, 0.5, in weight data studies effects and monkeys a the found. inhalation changes, in abnormal of Mechanisms thyrotropin-releasing hormone, exposure, death were Pertinent proliferative single and, that to b e 2,3,7,8- effects human. Animal. pigs of of it r e m a i n s associated with death lethality. results of loss subsequent suggest action, length understood, literature. Oral, liver which impending exposed by results the from activation in humans Inhalation. available clearly body weight of depletion, or Hepatic animals in reports exposure 4.3.2.3 cause anorectic of is n o t severe segregated results induced vitamin A TCDD the were 0.0049 of studies >0.5 feeding observed n g /kg /d a y, 0.00061 are not 46 available guinea pigs, et a l . 1979, NTP that represent a LOAEL of -0,001 Studies of (Kociba doses in studies 1982a) reviewed and mice by EPA (NTP (1985a) 1982a) ^g/kg/day in rats indicated for 2 years that are also hepatotoxic. Dermal, on humans other human. have been industrial studies, it is chemicals although also expected. be the some effects which Reports reviewed by symptoms 2,4,5-T case was more no firm in initial studied were to EPA the (1985a), have 2,3,7,8-TCDD. clinical study ages 6 to for elevated and alanine aminotransferase resided in slight, with group. (1982) reported the to major serum enzyme involvement, have in et cutanea al. that been the (1984) of tarda General discussion. of these detection the of have altered level of contamination, specific conditions factors, along with the the et urine al. of also acid levels liver e f f e c t s , as observed by persons Falk exposed et al. in Missouri — cutanea causative are pilot known who not have to which accident effects been subjects epidemiology failed to detect exposure. of have been 2,3,7,8-TCDD or the porphyria or the b i o a v a i l a b i l i t y of the contaminated nature of regard cutanea populations of site. and the Results to the tarda. studied, the 2,3,7,8-TCDD These in 2,3,7,8-TCDD in rodents. inconsistent with to investigated contaminated with enzymes ubiquitous may has The available. chemicals related (1986), tarda a g e n t s . The the M i s s o u r i tarda, exposure. and Chelsky chemicals serum may be the 2,3,7,8-TCDD other generally been liver inconsistencies were in Ideo They the Hepatotoxic to 3.6% and increase glucaric cutanea studies exposed to The accident. porphyria known hepatotoxicity studies the in a p o p u l a t i o n w i t h No These the or During in b o t h b o y s area. in porphyria reviewed by Jones the to The 2,3,7,8-TCDD. indicated by cases acid 122 been all animal. populations of enzyme exposed accident. 2 years. between however, study associated with diagnosed Falk as observed enzymes. No to b e Dermal, because levels, contaminated with exposed may have human microsomal epidemiologic reported porphyria of of hepatic of could be liver 1986). the compared after correlation one essentially effect serum al. Bogen as the h e r b i c i d e potentially et after years that study by 4.3% strong definitively were of to n o r m a l area -2.5 studies reported of gamma-glutamyltransferase were was a of contaminated Seveso a pilot Liver incidence returned levels of the activity been total Values (ALT) highly to and following levels there chemicals also (1979), to cause dysfunction" these were (Mocarelli D-glucaric in in Holden exposed were elevated urinary indicated by (1984) a the m o s t the 6 years who and the exposure 10 w h o serum (GGT) that is would exposed or b e e n "liver Since of accident examined yearly control (1973), and Vietnam veterans Seveso 2 years, May reported girls children exposure individuals in c h i l d r e n children were noted of and these inhalation routes contributed association'between complete 2,3,7,8-TCDD was route and In all contaminated with conducted the oral 2,3,7,8-TCDD 2,3,7,8-TCDD. predominant the of to herbi c i d e s may have (1980) in work e r s reports, made. A the effects exposed observed. (1979) , a n d H o l m s t e d t the that exposure b y In addition, chemicals, the h e p a t o t o x i c on populations contaminated with considered dermal, multiple conducted under confounding causative agents for 47 slight changes TCDD-induced a n d mice, in liver adverse 2,3,7,8-TCDD however, there response relationship; liver are effect have a are first observed no studies thus, pigs damage) at increasing difficult a doses as dose and to low other study supports is a by 0.1 in of on initial 2,3,7,8-TCDD In the investigated if 2,3,7,8- impossible. effects effects the dose- changes in other organ rats liver; in exposure induction et have case s - " t o segregate with cause the toxic response. (Beatty and Neal al . 1979, 1982). related Greig or the an systems compounds 4.3.2.4 to 1976) and for mixed has often toxicity Later for function of studies, appears rather demonstrated Schwetz in many than be th e DT- oxidase (Poland 1982, Poland and Knutson structurally related chlorinated with activity the Pertinent following available the relative (Poland ability and Glover of these 1973). Oral, human. No Oral, animal. studies review by (1985a) EPA durations Most guinea pigs. guinea pig Mg/kg/day As and differences, effects of sensitivity also (Dencker et M) (1984), this were and conducted with 8 weeks, to of animals were_not_ effective 0.04 indicates, the with a LOAEL of addition to the in mice, with thymus organ Thymus cultures to observed 3 x the from effects 10"® M, of cultures C57B1/6 toxicity of for Hartley 0.04 species mice, immunotoxic immunotoxicity in vitro which 2,3,7,8-TCDD on thymus the highest ranged ¿xg/kg/week immunotoxic fetal sensitive to in end point doses and Segregation very at observed for the exposures locus. 1985). mouse (1973) species, investigated indicated sensitive the A h al. DBA/2J al . a rats, in sensitivity been were no et ^g/kg/week.. I n differences in is as weekly 5 /xg/kg/week f o r 0.008 demonstrated and and minimum sensitive segregating with nonresponsive extensively and Knutsen of with been pigs, 2,3,7,8-TCDD have compared effects or available. has study by Vos a NOAEL of humans guinea the m o s t strain been responsive, 4 immunotoxic exposure and for mice the may be are rats, studies of between (ig / k g / w e e k regarding 2,3,7,8-TCDD in mice, toxicity. data inhalation literature. immunotoxicity * activity and 2,3,7,8-TCDD. the toxicity been well that function. rats Immunotoxicity in 1 suggested to it changes toxicity. enzymes animals has of enzyme 2,3,7,8-TCDD This toxicity in that acute microsomal liver study of enzyme in liver study makes studies the enzyme the this similar in chronic 2,3,7,8-TCDD and in of of correlates Inhalation. from that induction Kociba induce 2.3.7.8- TCDD found (1972) altered effects association between effect Observing and liver 2,3,7,8-TCDD-induced lack of laboratory 1979, In addition, dibenzo-p-dioxins of to of dose. of hepatic to demonstrated the severity al. however, diaphorase /ig/kg, indicator exposure Buu-Hoi reported mild observation TCDD was of (1983) guinea pigs the 2.3.7.8- 0 of resistant threshold sensitive associated with studies as a however, Early 1 that known toxic Collins increasing Additionally, has is n o t confirmation in h u m a n s adverse available it generally determine hepatotoxicity been and (a s p e c i e s subchronic mice, of made liver manifestation after Although Turner et have the clearly produces toxic only on been manifested. guinea in function, effects cultures are Ah ( E C 50 o f from the concentration TP -*0 1o 48 tested. in Effects thymus grafts, T and tumor lymphocytes. that received weeks of weight, 2,3,7,8-TCDD on the sensitization to antigens cell Thigpen as serum et demonstrated little al. as susceptible infection. This dose 2.3.7.8pups doses on using of thymic epithelium thymocytes. system Dermal, of chemicals of 82 human. There to results an (10.4% vs the in in humans altered immune performed point the in clinical animal number data and strain The exposed frequencies a T4/T8 not group, one on are ratio resulted is abundant most any reported that (11.8% 1.1%) vs also h a d non- of (8.1% <1.0 test an results excess suggest vs (12.6% of that exposure cell-mediated Immunity; out. animal the importance evaluated. response, will detect T-cell .... make data sensitive been very a specialized settings to study where _ available. the cannot be immune other abnormal in data have thus, immunotoxic or group of T-cell these be are of data, s h o w that and of The the the indicate end investigations of 2,3,7,8-TCDD-induced time fact analysis that toxicologic limited of onset, that not the evaluation of routinely assessment 2,3,7,8-TCDD produces of species, and most of available alterations regarding differences evaluating maturation of this end difficult. Investigation less in response in humans The in a altered (1986) energy will There immunotoxicity 2,3,7,8- the 155 u n e x p o s e d of have There may be in humans; those (1985), that failed and al. functional studies end point as in M i s s o u r i et carried 2,3,7,8-TCDD. and effective epidemiologic frequency studies discussion. the exposed depressed No of in 2,3,7,8-TCDD the a pilot associated with General of on In 154 Hoffman 6.8%), in the exposed animal. of to h e r b i c i d e s et of 11.8%). effects the corroborative duration a study increased abnormality Although for effect information exposed 2,3,7,8-TCDD. (35.3% vs Dermal, points little increased significant immunotoxicity this is Missouri, an 2,3,7,8-TCDD may be that the (1986) illness however, evidence al . Summit, The same inhibiting Stehr energy 8.5%). of signs effect e t al. by occurred, relative clinical studies gross exposure the exposure statistically vs provides cells 4 been reported Greenlee areas and test approximately study by for early 1982). from group had and al. subjects in Gray an also low-risk exposed 6.4%), in any and 40 the subset The in humans o f - I mmunotoxicity. In persons were a week bacterial in utero et not available. contaminated with 2.3.7.8- TCDD (Luster cultures, Longer-term have skin B but C57BL/6JFh mice once result following epithelium are n o t 2,3,7,8-TCDD high-risk signs cell the effects mice in adults. that subsequent did not lactation weight basis effects directly on immune effects and B6C3F1 decreases antigens, immuno g l o b i n s , and 2,3,7,8-TCDD from included susceptibility was Immunotoxic through a maternal acts the the rats exposure that produced TCDD that host exposure. of Fischer of to d e a t h 2,3,7,8-TCDD suggesting TCDD postnatal of (1975) 1 ag/kg more toxicity, system (bacterial development), were of immune response of the induced in dose-response in sensitivity; with studies regard these the have immunologic been immune system. relationships data w o u l d be to h u m a n h e a l t h . Also, effects concerned and of no with There are species assistance data are 4970 49 available that on the appears to immunotoxicity be very of sensitive 2,3,7,8-TCDD to other in m o nkeys, toxic effects a of species 2,3,7,8- TCDD. 4.3.3 Developmental 4.3.3.1 Toxicity Inhalation Pertinent data regarding following inhalation available literature. 4.3.3.2 No Animal. studies are toxicity, indicate 2,3,7,8-TCDD that administration. were Neubert and Dillmann et a l. on 2,3,7,8-TCDD al. and with kidney of 10 effects for have in E P A in the 1976, been effects In rats, and 1973). low No . 1973, as et Both of 1 pg/kg observed gavage effects al. 1976). (Courtney, after to 0.25 edema were pg/kg observed a number subcutaneously a (Sparschu Sprague-Dawley confirmed by only administration of -0.125 and gavage (Moore subcutaneous mice after developmental Courtney as (1985a), and rats observed at doses in N M R I administered been demonstrated exposure 2,3,7,8-TCDD. of demonstrated responsive developmental of to other that et al. of (studies at 0.3 rats. studies in summarized Weber et increases effects a l. al. of (1985) et al. or developmental and Dencker susceptibility cleft that indicates other to toxicological locus. 1985), or to and of mice additive exposure to palate end Additionally, 2,3,7,8-TCDD thyroxine An and Pratt o f mice; characteristic sensitivity simultaneous the and This the A h exposure 1986) the 2,3,7,8-tetrachlorodibenzofurans 4.3.3.3. to m a n y with 2,3,7,8-TCDD. for the susceptibility (1980) the treatment. (Bimbaum et in developed after simultaneous (Bimbaum 1986), developmental affect and Glover segregates polychlorinated biphenyls hydrocortisone compounds in addition 2,3,7,8-TCDD, demonstrated genetic differences mice kidneys toxicity, that both Poland genetic C57B1/6J and hydronephrotic and it specific the hormones triiodothyroxine to the effect was observed 2,3,7,8-TCDD and (TCDF). Dermal Human. human studied in mice doses organs and Ruddick respectively, toxicity by the al . gestation. internal concomitant (Lamb found 1985b). It h a s was et organogenesis of teratogenic points 2,3,7,8-TCDD not and cleft palate 1976), 2,3,7,8-TCDD was only kidney day These (1981) of were reviewed commonly at Khera pg/kg, most observed which in EPA the teratogenic been during 1971a,b; 0.03 is extensively Smith produced hemorrhage et toxicity animals studies, 1972, have exposure been these hydronephrotic anomalies 1976^-Smith and In mice, effects of or available. 2,3,7,8-TCDD has developmental single developmental of humans Oral Human. these the exposure The populations EPA (1985a) exposed to evaluated herbicides several epidemiology contaminated with studies of 2,3,7,8-TCDD. 50 It is assumed that the dermal; however, studies reviewed were reported cleft some elevated li p , incidence neural-tube talipes, stillbirths (EPA 1979, studies and in these cystic Hanify reviewed et al. to defects Thomas 1980, Dept, of Health New Zealand Aldred 1978, Smith et al. Bisanti et 1980). In al. 1982, of Vietnam veterans potentially in b i r t h palate, types and of noted have increases that the resulted result of active seemingly higher inherent studies, compounds, negative and teratogen nor The No studies a clearly of data be on exposure was 8 or which to similar that 1984). When and all authors defects factors. As may a geographic that provided investigations that Orange cleft other potentially studies these could 2,3,7,8-TCDD was critical in mice result is and the for the to be for of other chemicals data a al. 1973). the effects the monkey of with many time after 13 on days being uncertainties 2,3,7,8-TCDD data for sensitive the acute to of treatment experimental in w h i c h in epidemiology the with Additional of As that treatment et extremely that may effects, for adequate the determining whether indicated and with and results in humans. induction of lack of made for defects effects exposure, have inadequate animal the data from and to toxicants, (Neubert known exposed population, duration developmental toxicants, 2,3,7,8-TCDD, 2,3,7,8-TCDD on the guinea toxic is effects a potent agent. Reproductive 4.3.4.1 and exposure evaluate quantitative development 4.3.4 interpreting 1980, offspring in A g e n t risk, 1979, available. 11 p r o d u c i n g m a x i m a l ineffective regarding of induces developmental fetotoxic of defined levels developmental difficult 2,3,7,8-TCDD of are lack themselves pig, al . risk to the spina bifida, elevated exposure the h y p o t h e s i s discussion confounding presence days 2,3,7,8-TCDD individual birth review General quantitative data of risk concluded disprove Reggiani (Erickson et in al . 1977, study included for et al. 1978, no concomitant the et a l. other unidentified in Other in humans. Animal. 4.3.3.4 or (Nelson there was uncertainty the r e s u l t s , it w a s neither prove tumors 1979). control to that and increases correlation between McQueen et as and Kerr a 1980, case The that hypospadias well 2,3,7,8-TCDD exposed combined, from chance the correlation were a defects certain congenital defects as Field Bonaccorsi addition, detected to likely. studies i n c l u d i n g .cleft p a l a t e , demonstrate birth also correlation disease, 1981, exposure predominantly was abnormalities, kidney failed and possible defects, heart was exposure geographic of birth defects, and studies inhalation predominantly epispadias, similar exposure oral Toxicity Inhalation Pertinent data regarding in humans or animals available literature. following the reproductive inhalation toxicity exposure were of not 2,3,7,8-TCDD found in the 51 4.3.A.2 Oral Human. No Animal. following studies The observed doses et 0.01 exposure study (1979) 2 and reported of Fetal toxicity administration of and neonatal survival (1979) et without substantial increase not al . resulted generation. at The absence Nisbet statistically and weight, both and the Paxton the 0.01- (1982) to The monkey, 2,3,7,8-TCDD the of common however, either total of dose In (palatal abnormalities) an earlier diets Hg, fetuses feeding containing during pregnancy pregnancies TCDD or (Allen et in E PA the results Wistar which on rat It was et earlier androgens (1985) speculated reproductive pregnant exposure, pathways of to all by been data fetal increased Nisbet and the monkeys of the at and fetus toxicity 1985) are treated with abortion in and 1 of only minimal developmental dose of described gestation; were prior In both groups, a 4 test toxicity. maintained to total the indication ingested was 7 months Moore et account following Khera of 1.8 In on a n d 11.7 pregnancy two-thirds shown these to b e and of the al . for appear 2,3,7,8-TCDD in vitro not does to b e (Shiverick that may this male exposure, were a result of Levels of studied by treatment. participate estradiol affected by some 2,3,7,8- reduced (1973). rats levels Circulating effect (1985) the 2,3,7,8-TCDD d e p r e s s e d as to of male and Ruddick depressed hormone in males. rats the w e i g h t 2,3,7,8-TCDD-treated male however, studied might performance in exposure in suggested by organs dysfunction although chronic decrease observed by and that females, when in a It was reproductive al. 1979). (1985a), reproductive had been circulating Moore organs. the al . has the decreased (1984, either total effect was 2,3,7,8-TCDD. produced for This other were of eight monkeys (the to be an in abortion. reported effect ppt 0.1- fertility. level performance to 4 0 indication size, pg/kg/day dose sensitive there was The in analysis McNulty 20 pg/kg, no 500 of in rhesus days groups at fetal reevaluated The of Murray observed was index, at litter in any pelvis in a (1985a)] study generation. lower who doses. 0.001 levels renal more 2,3,7,8-TCDD in reproductive the or As diet 0.2 study, of ended over of effect this mice. abortion examined, 50 respectively) at reproductive or on decrease gestational appears tested, monkeys. t h e Fl properties rats 2,3,7,8-TCDD in dilated that toxic occurrence 1 /xg/kg o f lowest the than that of only (1982), 0 . 001-/ig/kg/day indicates very.__s e n s i t i v e effects concluded low dose the significant Paxton incidence and the as 2,3,7,8-TCDD S p r a g u e - D a w l e y rats. pelvises of and effects significant since statistically questioned by a considered effects, in d i lated renal observed in in EPA three-generation in adverse well bleeding have been [reviewed b y The survival, demonstrated as and vaginal studies dietary resulted Murray death been and mice) 9 ^g/kg/day. 0 . 1 /jg/kg/day also (rats that and /xg/kg/day dose 2,3,7,8-TCDD has in u t e r o (rats). in develop m e n t a l of between al. available. fetotoxicity short-term multigeneration are levels in 2,3,7,8-TCDD estrogen-metabolizing and Muther 1983) . 4973 in 52 A. 3.A . 3 Dermal Human. Epidemiology individuals with exposed 2 , 3 , 7 , 8-TCDD. predominant routes EPA route would (1985a) (1985), were plant Vietnam, and amounts not study, possible The females disposal, to 2 , 3 , 7 , 8 ;T C D D the that 2,3,7,8-TCDD lack of 2,3,7,8-TCDD exposure abortion conception. In of male exposure 2 , 3 , 7 , 8-TCDD. that have Reggiani to 1980) assess the on the the and indicate that 2,3,7,8-TCDD animals fully on chemical species that In is not of toxicant in humans no and et this is at the abortion. point a lack of high only of the the sensitive rhesus any dose level, which provides range and extent of potential evidence studies however, available species, monkeys, of humans speculated. Animal The with but only insufficient toxic are elicits toxicant; observed. the most sensitivity from which sufficient toxicant. in 1978, studies limited the [females al . 2,3,7,8-TCDD been only be et _ chemicals epidemiology have can to accident] reproductive to on the pregnant. that to the of potential populations do not provide been from exposed while a reproductive a powerful difficult female populations Bonaccorsi after been Although in is data time to soldiers, removed exposure limited 1980, sensitivity have addition, or m i c e sprayers, is during The resulting end equivalent al. they greatest temporally immediately is active. is organs the m o n k e y m a y b e species evaluate rats, it of absence conducted have in males there are reproduction; studied species. monkeys, limitations exposure spontaneous in animals this have effects concomitant herbicide studies the p r e sent in the reproductive and limitations, this this a observations that to is above indicate of large in humans. consistent with effects few exposure, exposure There period to p r o v e tests the (Bisanti adverse data adverse of in females in any of groups extent reproduction differences escape these the indicative concern, 2 , 3 , 7 , 8-TCDD, in the the of the extended periods Seveso effects Given only Studies in industrial of exposure of reproductive had contaminated with is addition, functioning of Orange result in male fetotoxicity of pre-mating and the v i c i n i t y application workers, through major been biologically occurred assess, particularly when to reviewed by a data plant w o r k e r s , whereas fetotoxicity studied as other available. that may have ^the p o s s i b i l i t y those the by discussion concerned mostly with of to for chemical time regard to b e and Nordstrom to A g e n t 2,3,7,8-TCDD produced although as are include of contaminated exposure which permitted from production plants. studies some exposed groups considered Forsberg exposed toxicity chemicals of on chemicals of male herbicide results evidence exposure that the limited by and which and and that is studies, interpret General route studies, soldiers performance, No been performed industrial and particularly with to A.3.A.A male or these in groups demonstrated Animal. other (198A) inappropriate reproductive is These employees, on reproductive the occur. of chemical clearly have dermal in Friedman to b o t h or the exposure conducted chemical accidents Although of also and studies to h e r b i c i d e s effects compared to a data in 53 4.3.5 Genotoxicity 4.3.5.1 Human Although 2,3,7,8-TCDD exposed to (1985a). in there chemicals Czeizel (spouses and and Mottura al. been no are studies a number contaminated with and Kiraly aberrations (1976) of human of of soldiers) involved 1981, have, however, individuals exposure to studies was (DiLemia a satellite decrease et ribosomal a l. of 4.3.5.2 A were recent review by activity of to genotoxicity the in mixed that human exposure to 1983, the which 1980, Kaye et Some latter is evidence subjects decreases represent on to determine (Mulcahy (Reggiani in similar data in an soldiers consistent with effect may possibly All inability In in v i v o positive obtained damage to chromosomal other potentially quantitatively this activity with are mice) al. the (1972) and cell death cytogenic have produced positive depended posttreatment the toxic locus, on sampling and positive et al. administration of responsive, C57B1/6J, sampled at 2 4 h. There mutagenicity of 2,3,7,8-TCDD are 2,3,7,8-TCDD to of Seiler (1973) the The are and possibly tests from in yeast in host-mediated (Bronzetti al. (1982) time, with results 96 et al . observed negative h 2,3,7,8-TCDD usually (1985) observed negative 2,3,7,8-TCDD limited of a n d 4.3. at h e p a t o t o x i c and n o n r e s p o n s i v e , DBA/2J, also from et the results 4.2 results Loprieno properties Meyne The intrasanguineous assays, genotoxic for testing would be in Tables and responses the and evidence certainty. summarized test cytogenetic after is additional In vitro the A h of but extensive Although binding the m u t a g e n i c there from segregate with to b o t h of that test material. 24 h in v i v o (1986) of H u s s a i n et posttreatment. responses Giri studies standard plate the level noted exposure EPA aberrations. skin eruptions (SA ) , of al. 2,3,7,8-TCDD-exposed regions. resulting the (using CD-I results when et chromosomal association 2,3,7,8-TCDD, results artifacts impurities levels Orange accident Tenchini in increase from workers to A g e n t chromosomal authors concluded demonstrate early positive that had an 2,3,7,8-TCDD. 2,3,7,8-TCDD required 1983). the to nonhuman assay in confounded by and 1982, reviewed Studies to Nonhuman genotoxic both The organizing exposure of likely genes, 1982). active^, c o m p o u n d s effect in only as exposed populations there was plant. Seveso to d e t e c t studies after x-irradiation aberrations al. failed these exposed The the n u c l eolar extent in et that lymphocytes in the 2,3,7,8-TCDD. functional observed DiLemia on human 2,3,7,8-TCDD, reported of peripheral individuals e t al. 1985) the children of populations studies European 2,4,5-trichlorophenoxyethanol 1980) of have chromosomal East of there alone, supportive observation data in v i v o liver macromolecules of (Poland mice for the the low- and Glover 1979). 4.3.5.3 The General nonhuman discussion genotoxicity negative results were reported positive results were observed, data on in m a n y the 2,3,7,8-TCDD are of systems, the response assay was conflicting; and generally weak. when These I SÄ, % Table 4.2. G enotoxicity o f 2,3,7,8-T C D D in vitro 1, End point Species (test system ) G ene m utation S a lm o n e lla ty p h im u riu m Results with activ atio n /w ith o u t activation M cC ann 1978, G ilbert et al. 1980, G eiger and N eal 1981, M ortelm ans et al. 1984 (reverse m utation) S. typ h im u riu m N ot te s te d /+ Hussain et al. 1972, Seiler 1973 N ot te s te d /4- H ussain et al. 1972 (reverse m utation) 1 E sch erich ia co li (reverse m utation) S a cch a ro m yces cerevisia e + /~ (reversion) t Rogers et al. 1982 + / - Bronzetti et al. 1983 + /N A " Bronzetti et al. 1983 Chinese ham ster cells (sister chrom atid exchange) N ot te ste d / — T oth et al. 1984 Baby ham ster kidney cells BHK C 3 H / I 0 T I / 2 cells , N ot te s te d /+ H ay 1982 N ot te ste d / — A bernathy et al. 1985 S. cerevisiae (gene conversion) S. cerevisiae (host m ediated) Cell transform ation Bronzetti et al. 1983 N ot te s te d /+ , and not te ste d / — L 5 I78Y mouse lym phom a cells (forw ard m utation) C ytogenetic References "N ot available. CD -3 I ) \ 1.» ■ Table 4.3. G enotoxicity o f 2,3,7,8-TC D D in vivo End point G ene m utation Species (test system ) Results D ro so p h ila (sex-linked References Z im m ering et al. 1985 recessive lethal) C ytogenetic — Z eiger 1983 — Z eiger 1983 R at (sister chrom atid exchange) — Lundgren et al. 1986 R ats - m arrow cells (stru c tu ra l aberration) — G reen and M oreland 1975 R ats - m arrow cells (stru ctu ral aberration) + G reen et al. 1977 M ouse - m arrow cells (stru ctu ral aberration) + Loprieno et al. 1982 M ouse - m arrow cells (sister chrom atid exchange) — M cyne et al. 1985 M ouse - m arrow cells (stru ctu ral aberration) — M eync et al. 1985 D ro so p h ila (sister chrom atid exchange) D rosoph ila (stru ctu ral aberration) M ouse - m arrow cells (m icronucleus) tO' a»!JE « Ä. I M eync el al. 1985 " Ln Ui 56 conflicting 2.3.7.8- data TCDD difficulties the h i g h may arise toxicity anticipated to result a genotoxicity Sufficient data of extent the of are exposure. of after the not studies the optimal the the observing studies in humans, A . 3.6 Pertinent data these exposure cells data a effective not a define the does or genotoxic Paustenbach genotoxic the the opinion. on in these duration. examined may Because cannot be used of to the studies not provide effects. et agent. toxicological of lack of data exposure were genotoxic regarding available literature. A.3.6.2 Oral, Animal. route. The bioassays of animals is relevant a was good In addition, not have the been limitations demonstrate of that hazard. proven daily study. to b e summarized The A.3.6.3 to Other an animal Although there 2,3,7,8-TCDD, herbicides or in studies study by found because study by in the used et al . studies carcinogen the et al. oral key numbers (1978a,b)- compound was gavage et al . (1978a,b) support presented as in the (1978a,b) carcinogenic which are the are sufficient Kociba the all ha v e via studies Kociba of and animals than biweekly by the K o c i b a oral not conducted, (1978a,b) indicator studied. The In are predominant known effects potentiated the contaminated that study the has potency of are conclusion in Table that have b e e n have contaminated with route inhalation 2,3,7,8-TCDD was any cohorts cohorts exposure some compounds, and by of chemicals although addition, no a number industrial probably, dermal, likely. sensitive of the rather the 2,3,7,8-TCDD were A . 5. Dermal Human. solely been al. scenarios diet A.A. is have et of animals available. Although both details in Table 2,3,7,8-TCDD the or carcinogenic durations, Furthermore, the m o s t 2 . 3 . 7 . 8 - TCDD. is and Kociba exposure in are of bi o a s s a y s exposure to h u m a n carcinogenicity studies 2,3,7,8-TCDD. administered the of humans 2,3,7,8-TCDD (1982a) and (1982a) No number that NTP of exposure human. A demonstrated been the and from nonhuman differences which Testing compound Inhalation inhalation that of that for is limited by indication of extent this discussed by these sk i n lesions, pose as testing which would be data to p r e c i s e l y resolve of systems, 2,3,7,8-TCDD cytogenetic not following NTP test in activity. Carcinogenicity A.3.6.1 _ and only exposure for that primarily gross does insolubility investigators, available The of either 2.3.7.8- TCDD in some difficulties of b i o l o g i c a l largely negative some are technical a lack extreme concluded development time the 2,3,7,8-TCDD estimate from from in a v e r y n a r r o w w i n d o w of assays, have Human from result a l . (1986), mechanism than observed doses. As result rather and only been itself. to 2,3,7,8-TCDD have these oral a minor observed may have compound for exposed been exposed cohorts exposure was contaminant either caused was also of or these 57 Table 4.4 Sum m ary of the oral carcinogenicity bioassay of Kociba et al. (1978a,b) A nim al Sex Dose tested (M g/kg/day) SpragueDawley rats M Control Squam ous cell carcinom a of the tongue, adenom a of the adrenal cortex, and squamous cell carcinom a of the hard palate 0 /8 5 0.001 Squam ous cell carcinom a of the tongue 1/50 0.01 Squam ous cell carcinom a of the tongue A denom a of the adrenal cortex 1 /5 0 0.1 F Tum or type Squam ous cell carcinom a of the tongue A denom a of the adrenal cortex Squam ous cell carcinom a of the hard palate Incidence 2 /5 0 3 /5 0 5 /5 0 4 /5 0 Control H epatocellular carcinom a 1/86 0.001 H epatocellular carcinom a 0 /5 0 0.01 H epatocellular carcinom a Squam ous cell carcinom a of the hard palate 2 /5 0 1 /5 0 0.1 H epatocellular carcinom a Squam ous cell carcinom a of the hard palate Squam ous cell carcinom a of the lung 11/49 4 /4 9 7 /4 9 4979 50 fD O Table 4.5. Other oral studies supporting the conclusion that 2,3,7,8-TCDD is an animal carcinogen M ethod of exposure A nim al Sex/ num ber Doses tested T um or type References Diet SpragueDawley rats M/ I O 0 .0 1 ,0 .0 0 5 , 0.05, 0.5, 1.0, or 5 ppb Increase in total tum or incidence Van M iller et al. 1977a,b G avage O sborneM endel rats M /5 0 0.01, 0.05, or 0.5 /ig /k g /w eek Follicular-cell adenom as and carcinom as o f the thyroid N T P 1982a O sborncM endel rats F /5 0 0.01, 0.05, or 0.5 /ig /k g /w e e k N eoplastic nodules and hepato­ cellular carcinom as of the liver N T P 1982a B6C3F1 mice M /5 0 0.01, 0.05, or 0.5 /ig /k g /w e e k H epatocellular carcinom as N T P 1982a B 6C 3FI mice F /5 0 0 .0 1 ,0 .0 5 , or 0.5 /ig /k g /w c c k H epatocellular carcinom a and follicular-cell adenom as of the thyroid N T P 1982a Swiss mice M /4 4 0.007, 0.7, or 7.0 /jg /k g /w c e k H epatom as and hepatocellular carcinom as T oth et al. 1979 4980 59 EPA (1985a) to h e r b i c i d e s al. (1980) between and and 1981; between and exposure al. and and et al . 1981; Johnson et Wolfe et al . al . al. 2 . 3 . 7 . 8- T C D D in Smith of sarcomas of Cook (Ott and of et 1981; et an EPA et studies Hardell et al. association sites) (1985a) and - and Hiremath al. also cancer. 1982; Zack Smith Fingerhut The is 1980; did not epidemiology exposure al. and Pazderova-Vejlupkova 1985). (1987) the other (of vari o u s exposed Axelson an association between herbicide cancer al. by association 1981; reported reviewed by and Pearce et an of 1979, 1986) Riihimaki exposure some al. found series al. of humans studies not et et al. a l. detect Suskind et al. 1983; (1984), an extent of exposure studies is questionable, available for Cook association all to of and the . Animal. tumors et detect 1981; quantification studies. et 1985; and Ott between herbicide precise to 1980, 1984; (1986), and a however, induction Cook cancer, Palovi studies (1977) Eriksson soft-tissue failed 1980; et 1979; studies, the epidemiology 2 , 3 , 7 , 8 - T C D D . The Frentzel-Beyme stomach 1985; Other (1986), exposure several and Sandstrom Lynge lymphomas. et Thiess exposure (Hardell 1980, reviewed contaminated with in There is laboratory only limited animals administered 2,3,7,8-TCDD mice^-Fiad a n increased evidence following alone to incidence Swiss of that dermal 2,3,7,8-TCDD produces exposure. mice. Female fibrosarcomas in NTP (1982b) mice, the but not male integumentary system. As reviewed by 2 . 3 . 7 . 8- T C D D al . (1978) in CD-I Slaga a failed to detect mice skin 2 . 3 . 7 . 8- T C D D in affect mice homozygous the DMBA however, these groups (1982b) ability that of but also 2,3,7,8-TCDD DMBA produced mice for could only be hyperplasia indicated (Berry Pitot single (DEN) et TPA but 2,3,7,8-TCDD associated with 2 . 3 . 7 . 8- T C D D has that DNA binding the h airless promoted by mice, of as trait not did not skin a promotor. the such TPA, as 2,3,7,8-TCDD known and also C o h e n et demonstrated promotion by intragastric dose of followed by repeated 1979). Although the h e p a t o c a r c i n o g e n subcutaneous Even in incidence whereas not a in tumors the observed suggesting Other tumor also In HRS/J type), skin that studies could block prevent 1979, al. (both TPA; commonly action. not in CD-I 2,3,7,8-TCDD tumors 2 , 3 , 7 , 8-TCDD. of precludes however, differences (and w i l d pretreatment with al . (1982), produce mechanism carcinogens DMBA promotion with many to 2 , 3 , 7 , 8 - l'CDDT experiment, only as of and incidences with DMBA-TCDD act mice, tumor the k n o w n p r o m o t o r promotors different ability Webster (1980) et al . a the al. the subsequent initiation dermal study, administering a diethylnitrosamine injections of et that examined genetic to trait, as p r o m o t i o n w i t h homozygous that indicated promoting treatment et Berry 2,3,7,8-TCDD weak similar the Poland and multiplicity) heterozygous "in by or v e r y Swiss had that skin. ( DMBA), and data skin painted with the hai r l e s s initiation with alone also in followed by results." promotion, for NTP activity tumor promoter 2,3,7,8-TCDD included demonstrated mice a first with of the activity unpublished promoting skin. as concluded, interpretation after reported mice act treated to h a v e tumor-promoting evidence to that treated with only any applied dimethylbenzanthracene animals (1982b) conflicting initiated with animals failure is (1985) Senear to there tumor promotor wh e n either had no 2 . 3 . 7 . 8- T C D D NTP (1985a), as and N e snow activity EPA acts 2 , 3 , 7 , 8-TCDD. No 60 hepatic tumors but combined the were observed treatment in animals resulted given in the DEN or 2 , 3 , 7 ,8-TCDD development alone, of h e p a t o c e l l u l a r carcinomas. 4.3.6.4 General The evidence carcinogenic is poorly is documented materials. tissue sarcomas the data and sites and data, however, clearly although there some international regulatory carcinogen rationale response for of describing lack of promotion strong evidence assays above) and stage model would support 4.4 INTERACTIONS There chemicals. in rat are As rate or fate of toxicity The only the A) et on the hence, and (1985a), prior this al . al . is interactions 1986). interactions study of et al. of mice of With (Shu et al. 1979, acts on of results The Cohen et changes in the a twohowever, with other strong that are observed inducer 1979) altering alter either inhibition tumorigens al . in (as 2,3,7,8-TCDD will of known poor the carcinogen. is compounds a The the assays, 2,3,7,8-TCDD to and as 1987). is b a s e d complete is scientific 2,3,7,8-TCDD (1980) system. enzyme that have and may the in be — the an metabolic been 1985, to regard 2,3,7,8-TCDD observed are Bimbaum on the induction et al. 1985), 2,3,7,8-TCDD-induced to and the similar polychlorinated the h o r m o n e s is u n c l e a r . the 2,3,7,8-TCDD response tumors 2,3,7,8-TCDD in study bioassays, a to compound. 2,3,7,8-TCDD (Weber that using al . many enzyme tumorigenic another stomach et exposure of regard technique interactions toxicity (Berry et sensitivity latter 2,3,7,8-TCDD in EPA in and positive feeding data co-administration (Lamb Pitot soft- of workers skin painting and polychlorinated biphenyls in mice increased of classic of studies. a promotor DNA, OTHER CHEMICALS of of to WITH the other effect dibenzofurans palate that With groups to w h e t h e r as potentially questioned whether increases indicate classic 2,3,7,8-TCDD-induced and additive of The skin bioassay example the study activated by of two is 2,3,7,8-TCDD induction appropriate. the to short-term mutagenicity for b i n d i n g the enzymes; 2,3,7,8-TCDD mouse the few in m a n y liver. of metabolism detoxified by 2,3,7,8-TCDD discussed been disagreement as 2,3,7,8-TCDD other the a carcinogen promotor using the v i e w of microsomal the as 2,3,7,8-TCDD vivo described or community to extensive carcinogenic, complete is that exposure from similar in other more is is it has cancer, small, (1) occurred evidence in stomach reported animal exposure sites, studies because from various relatively not been (2) strongest increase studied were The and The epidemiology to a s s e s s at various tumor observed have from human difficult active combining & discussion thyroxine environmental the high and cleft cleft the palate with triiodothyroxine exposure, levels of and the of hormones relevance used in 5. 5.1 MANUFACTURE, 2.3.7.8impurity TCDD States. during only use is methods are wastes. Some It the trichlorophenol is is n e i t h e r is synthetic available of the for the a fungus, situ 2.3.7.8- TCDD (1) and (2) dichloro-substituted commercially undesirable of small for which At the of imported amounts as into an 2,4,5- present field-tested time, its and untested 2,3,7,8-TCDD-containing are incineration at high the aid of a catalyst, destruction stabilization cementitious the of in the (in c ase and in during is a on a presence of asphaltic laboratory dichlorocatechol halogenation manufactured product to synthesized derivative 2,4,5-trichlorophenol Releases is condensation dichlorobenzenes of soils) materials. (EPA of of but is by it s is n o t produced compounds intermediate primarily or 2,3,7,8-TCDD States manufacture synthetic scale substituted dibenzo-p-dioxin 1985b). the U n i t e d the with (see as an for w h i c h Sect. 6.2 on Environment). IMPORT 2,3,7,8-TCDD is not imported into the United States (EPA 1984b). USE 2,3,7,8-TCDD control against it h a s probably purposes. At has been insects never been present it tested for flame-proofing polyesters and wood-destroying is fungi commercially produced only used as a in Germany; or research used for chemical and as a however, these (HSDB 1987). DISPOSAL For the incineration of >30 dissolving disposal at as the found an containing methods to b e ruthenium were either not shown residues of 800 satisfactory. in water, field-tested soils have and temperature impurity have material oxidizing with Satisfactory of wastes a minimum s was 2,3,7,8-TCDD and and in "''PRODUCTION processes: time Several photochemical addition in compounds destruction with through the of disposal substrate, 5.5 DISPOSAL commercially manufactured nor intermediate. research. a hydrogen-donating 5.4 AND inadvertently promising methods oxidative biodegradation by produced manufacture a in chemical temperature, 5.3 USE, OVERVIEW the U n i t e d 5.2 IMPORT, been or are 61 2,3,7,8-TCDD, and a contact Polychlorophenols of b y catalyst the until or 1 2 0 0 #C at disposal containing dispersing a nonnucleophilic for available promise to disposed tetraoxide methods containing organic -70°C of (HSDB or solvent, 1987). 2,3,7,8-TCDD- recently. The presently used following for the disposal two 62 of 2 , 3 , 7 , 8-TCDD: cementitious and c h ryso sp o riu m , the in thermal (1) stabilization asphaltic a white rot destruction of the presence of 1986). A method (similar 2,3,7,8-TCDD with soil discusses the area fungus; a mobile disposal potassium hydroxide of (3) 2 , 3 , 7 , 8-TCDD; polyethylene to m e t h o d of the (2) in the p r e s e n c e from the alkali through materials; of the 4) olive Seveso situ and addition with system ultraviolet photolysis glycolate oil reagents (a h y d r o g e n accident. A et al . (des for Rosiers destruction donor) recent a mobile (Rogers of P h a n ero ch a ete incineration (4) for p h o t ochemical 2,3,7,8-TCDD by destruction unit in degradation was of used report poly(ethylene)glycol1987). 6. 6.1 OVERVIEW The important production and of municipal sources use and of industrial produced T, related herbicides, and The fate of It appears during photolysis and may of 2.3.7.8- TCDD be in surface '■’ f a c t o r 7900 sink of to of movement of As occur soils in in the is observed 2.3.7.8- TCDD 1 to low 3 years immobile may soils. Although not of plants underground as in soil, but the occur accumulated, aerial is is > 1 to year, The and TCDD is during movement may estimated half-life and the 10 to level the same of plants 12 of years in TO Although the the THE 2,3,7,8-TCDD order of contain 50% lower following paragraphs levels are 6.2.1 Production The to the sources and Use of The 2,4,5-T has been in commercial 2,4,5-T Orange, a reduced 1970, Hexachlorophene, and pentachlorophenol no of 2,3,7,8- and Chlorophenols were of background to <0 . 1 to 1960 c o n t a i n e d up in c o m m e r c i a l p g /g , a n d m o s t p g /g 2 , 3 , 7 , 8 - T C D D . contain <0.02 esters 0.02 to 2,3,7,8-TCDD years of butyl 54 manufactured of 2,4,5-T p g /g from and 2,4-D 2,3,7,8-TCDD. trichlorophenol, contains 2,4,6-Trichloro-, 2,3,4,6-tetrachloro-, found 2,3,7,8-TCDD. ng/g produced prior level contained 2,3,7,8-TCDD. concentration <1 samples of Certain Herbicides today may germicide to but ng/g a 0.2 isomers sources its recent available 1:1 m i x t u r e produced before 0.5 the for 2,4,5-T 2,3,7,8-TCDD. Agent discuss responsible clear. phenoxy herbicide fig /g 100 ENVIRONMENT environment, not of the ------- RELEASES in the but horizontal concentrations. 6.2 is remove The bioconcentration vertical The of parts likely in runoff water surfaces in parts transported 2,3,7,8-TCDD and photolysis. soils, minimal on of undergo half-life (P im e p h a le s p r o m e la s ) matter. interior The are sediments. organic soil understood. can be water in most Italy, chemical air m a y airborne that surface absorbed magnitude of vaporization is of clearly the 2,3,7,8-TCDD fathead minnow Seveso, in deposition. sink in 2,3,7,8-TCDD in containing is are are chlorinated benzenes. processes 2,3,7,8-TCDD 2,3,7,8-TCDD soil-bound flooding. soils 2,3,7,8-TCDD aquatic 2,3,7,8-TCDD 9300. two disposal is n o t dry that ultimate The and and environment 2 , 4 , 5 - t r i c h l o r o p h e n o l , 2,4,5- 2,3,7,8-TCDD such The waters. of the chlorophenols, incineration improper of environment is air. from water estimated half-life ultimate the removed by wet the in and hexachlorophene, and in 2,3,7,8-TCDD distances sediments wastes, and particulate-bound atmospheric long 2,3,7,8-TCDD the m a n u f a c t u r e 2,3,7,8-TCDD that of certain herbicides wastes of ENVIRONMENTAL FATE to contain <0.1 2,3,7-,8 - T C D D w a s (2,3,7,8-TCDD detection p g /g other detected at limit 0.03 of tetra a ng/g) in all sodium p e n t a c h l o r o p h e n a t e , 2,3,4,5-t e t r a c h l o r o p h e n o l , and 4385 63 64 hexachlorophene. 2,4,5-Trichlorophenol, on to 6.2 found /ig/g 2 , 3 , 7 , 8 - T C D D . S i m i l a r l y , to contain other 1985b, HSDB 1987, 1985). From the Czuczwa tetrachloro Rappe 1984, analysis of and Hites (1986) facility. The 2.3.7.8- TCDD 6.2.2 The concluded and are tetra other subsequent Thermal Small reaction (Rappe 1984). of Lower concentrations in of 38 salts analysis Lakes, flue concentration also of and gases of fg/g. from and Hites (1984, unable to limit 1.2 n g / k g (HSDB 1987), (EPA 1985b). /ig/kg 2 , 3 , 7 , 8 - T C D D gasoline were but coal Accidental fires 6.2.4 such a Improper chlorine- been detected and but also air industrial wastes release at that and of woods or 0.3 tetra at ng et transformers to is Office the analytical a detection did produce (Marklund State of the 0.65 powered with 2,3,7,8- in e x h a u s t s 2,3,7,8-TCDD the Great source automobiles to was Upon the other burning detected maximum level chlorophenoxy the from the 1983). although contain <0.05 gases containing and Barnes Saginaw River, gasoline flue ground 2,3,7,8-TCDD capacitors contamination 1984, from the incinerators, in ambient Exhausts to in determined produce of Disposal disposal of of Chlorinated certain of al. 1987). containing environment. Building An in chlorinated benzenes, leading to Falls, the in environment. contamination New York, detected. of sites Similarly, contaminated with up to etc., Examples the where of such in during the related herbicides, may be a source improper are 2,3,7,8-TCDD sites produce^ and environment several 1750 Vastes chemical wastes the Niagara Chemical 2,4,5-t r i c h l o r o p h e n o l , 2,4,5-T, 2 . 3.7.8- TCDD were not 2,3,7,8-TCDD were 2,3,7,8-TCDD hexachlorophene, /ig/kg w a s to N e w York. Improper manufacture does 1,2,3,6,7,8-hexa- incineration, unleaded involving chlorobenzene will Binghamton, of concluded 2,3,7,8-TCDD was powered with of found dibenzo-p-dioxins 1979). Bay, did not reported no automobiles example been process (Buser (Rappe separate of km , this of higher municipal 1986) was Combustion leaded distinguish has experimentally of Saginaw isomers. TCDD/24.8 production substituted polychlorinated benzenes, 2,3,7,8-TCDD tetrachlorodibenzo-p-dioxins of five Incineration sediments used was the 2,3,7,8-TCDD esters, produced Czuczwa method site, of through photodechlorination amounts 2,3,7,8-TCDD have From 2,4,5-T source Reactions incinerators. ethers Asshauer Ontario could not photodechlorination of both amounts as likely of phenoxy herbicides dibenzo-p-dioxins from municipal estimated and Lake up isomers. 1,2,3,7,8,9-hexachlorodibenzo-p-dioxin average the c o n d e n s a t i o n •r e a c t i o n s ; h o w e v e r , the (EPA Weeren however, dibenzo-p-dioxins. Tr^ce from 2,3,7,8-TCDD a western formed during photodechlorination observed no were Reactions 2,3,7,8-TCDD substituted 6.2.3 but contained herbicides a pentachlorophenol from photochemical also was o ther hand, ether 1986, of that m e t h o d used, polychlorinated produce Hagenmaier analytical Photochemical produce isomers sediments tetrachlorodibenzo-p-dioxins the diphenyl up the /ig/kg 2 , 3 , 7 , 8 - T C D D the to of disposal Love a state Canal, level of 672 of Missouri (Tiernan et al . 65 1985). 6.3 ENVIRONMENTAL FATE The with fate of certainty. recent years data gap to exists 2,3,7,8-TCDD Although some elucidate its in this present predominantly may remove and wet of In gas phase. the atmosphere from deposition. air, Even is n o t experiments with half-life of atmospheric gas-phase 2,3,7,8-TCDD. can be environmental sediments 1986, of sink of airborne waters The biodegradation two processes volatility that may and In natural substrate waters, is a 2.3.7.8- TCDD sediment process of In TCDD year. to in this et al. is > 1 . 5 The shown is with soil not leach A In sink of the air. The is The than 2,3,7,8-TCDD a is ultimate likely to b e and Hites amounts al. account was not tri- has due on not to a its of provided loss its of sorption on d e c h l o r i n a t i o n _____ dibenzo-p- estimated half-life the 2,3,7,8-TCDD is the factor for basis (EPA in most soils a wet weight of estimated half-life sediment. (P im e p h a le s fathead minnows the b i o c o n c e n t r a t i o n 9300 for dichlorinated the aquatic of studies observed alone, with are 2,3,7,8-TCDD degradability ecosystem the of of hydrogen-donating may 1983), that The photodegradation. photochemical model slow. 2,3,7,8-TCDD suspension appreciable lake water of of photolysis lake water, is expected through of very soil, organic been shown significantly Force solvent rot soils, degrading microorganisms. remove some surfaces 2,3,7,8-TCDD can be greatly cm in 12 y e a r s Although is p o s s i b l e or biotic fungus 2,3,7,8-TCDD. in n a t u r a l 10 carbon content A white degrade immobile of Base. leaching low organic invertebrates. to to b e downward movement soil soil years. from Eglin Air solvation with may the days. higher is p r o b a b l y and to gel, few Czuczwa removal formation that a 2,3,7,8-TCDD 1985b, Adams 1986). 2,3,7,8-TCDD other 1986, is u s u a l l y 7900 that of silica the photodegradation experiments have et to p h o t o l y s i s ultimate species the balance sediment-containing rainfalls. soils the flow-through p r o a e la s ) and The the small Matsumura attributed leading 2.3.7.8is > 1 1982, in of photosensitizers through mass and biota. dioxins. Recent al. coated on fast process, provided by et to b e processes atmospheric 2,3,7,8-TCDD showed no 2.3.7.8- TCDD evidence the atmospheric Although however, (Tsushimoto of the b a s i s in w a t e r for observed photodegradation; definite On distances 2,3,7,8-TCDD the p r e s e n c e as likely 1982). solvents in water, of in are photochemical and Hites important the p r e s e n c e is directed substantial important estimate particulate in d i s t i l l e d w a t e r or a 2,3,7,8-TCDD may be long photodegradation. in hydrogen-donating 2,3,7,8-TCDD of be two understood been 2,3,7,8-TCDD may be (Eitzer and Hutzinger an lifetime transported surface Choudhry The The 2,3,7,8-TCDD particulate it is n o t have 2,3,7,8-TCDD available. atmospheric that and soil efforts in d i f f erent media, of such j fate half-life particulate water, area. 2,3,7,8-TCDD photochemical air, in the 2,3,7,8-TCDD degradation half-life in experimental as 2,3,7,8-TCDD in rare a This of does from 2<3,7,8-TCDD earthworms biodegradation of Both volatilization soil usually or other (P h a n e r o c h a e te c h r y s o s p o r iu m ) probably because from irrigation observed instances result mixing by by was surfaces. enhanced by the the does not lack of has occur this or and photoreaction The presence photoreaction on of hydrogen- 4987 £ 6 ? ^ 66 donating substrates photoreaction will (e.g., be estimated half-life but the 1985b, half-life Freeman of in and is will absorbed by accumulated. the interior Schroy 1986, level as soil, on higher plants or arachis beyond on of the soil soil Bumpus leaves photolyze with The oil 2,3,7,8-TCDD 2,3,7,8-TCDD present herbicides olive insignificant et surfaces may be al . 1985, of plants a half-life and is oil) of probably but the aerial part 1982, contains Sacchi et a l. 10 soil. soil is to 1 to The 3 years, 12 y e a r s HSDB (EPA 1987). a result a few hours. The chemical translocated, but it the same -50% may be lower of at spraying is n o t concentrations 1986). / CY* The layers. as absorption by underground parts (Choudhry and Hutzinger in surface 7. 7.1 POTENTIAL FOR HOMAN EXPOSURE OVERVIEW The concentration 2.3.7.8- TCDD alone) fg/m^ (femtograms isomer constitutes 2.3.7.8- TCDD of incinerator has fire per 0.23 to the a ppm 2,3,7,8-TCDD was and leachates from In urban to 0.009 are contaminated by the contaminated by 2 . 3 . 7 . 8 - TCDD. waste Fish States, or in contributor exposed level of United many detected in and persons in of ranges tissue to 7 pg/g. Values as h i g h individual heavily exposed during the methods the of 2,3,-7,8 - T C D D h a s control 0.6 in population pg/g, and sites residing Although of near these estimated per kilogram at are sites 1.3 may no be municipal daily human of b ody to 3.3 fat to in in the United is the it has c h i e f ---- exposed in human milk with exposure levels, susceptible incinerators to 67 may 0.02 of 1.9 workers population exposure to residing near also 2,3,7,8-TCDD, ( p g / k g ) , is general to h i g h e r nursed by mothers of a m e a n value a mean value the in an sensitive data regarding and a South European disposal in the with from using more with tissue The detected in ■ been adipose pg/g, in h u m a n m i l k pg/g, of No population 20 operation pg/g, of 1750 ng/g determined. pg/g were 2.3 soils in other exposure weight crawfish has b e e n reported to in selected Michigan general however, trace range spillage to in humans, 99 in most in human mil k waste Breast-fed babies as detected at found chemical controlled The Sweden also there improper 2 . 3 . 7 . 8 - TCDD. improperly was the spraying States; 2,3,7,8-TCDD in in Germany 2,3,7,8-TCDD countries. not been the U n i t e d of measurement, general pg/g. at areas up a been 15 effluents in Missouri 2,3,7,8-TCDD in the and Since from undetectable of and and samples. 2,3,7,8-TCDD Levels and the ever been 2,3,7,8-TCDD. in Missouri have adipose Canada soybeans, mean value Vietnam. 5 of in oils, lakes in containing detected Stables 67 p g / g and pork investigators. 2,3,7,8-TCDD from 2,3,7,8- analytical is 2,3,7,8-TCDD had to rice, of been waste Shenandoah levels of 2,3,7,8-TCDD 2,3,7,8-TCDD wastes, a municipal industrial limits of transformer level Silvex of levels have collected chicken control of of 2,3,7,8-TCDD has detection from the b o d y b u r d e n and States no containing samples Canadian to analyzed by sites, 92 The 2,3,7,8-TCDD certain concentrations the stack a much higher than level contained undetectable 2.3.7.8- TCDD was of oil the accidental of to concentration fg/m^. fg/m^. The 18 2,3,7,8-TCDD air around Other certain hazardous the to 4 . 6 application of higher soil 0.9 concentration dump The Much 2,4,5-trichlorophenol. A river 38 the below soils, ng/g. The pg/m^. chemical soils the as after in d r i n k i n g w a t e r . uncontaminated in that Indiana, tetra.isomers, the produced pg/m^. 0.62 was Assuming total New York, field (not in Bloomington, air would be estimated 0.47 surrounding methods. of tetrachlorodibenzo-p-dioxin air meter). 2,3,7,8-TCDD been air <0.0002 cubic 3% in Binghamton, reported total ambient in Bloomington concentration T C D D — at of in through be at higher expressed as inhalation, risk. picograms 0.5 to 'P-SolO 68 5 through fish. of the The 850 mL and the human intake LEVELS 7.2.1 of milk, exposure and a milk is 20 half-life of 2,3,7,8-TCDD of 5 -kg b a b y to 2,3,7,8-TCDD has MONITORED OR 200 The level of 2,3,7,8-TCDD is b e y o n d concentration of and particulate the total phase an 92 the air Since municipal in flue the area TCDD, there level these compounds of the are publications isomersT~The levels incinerators are 0.05 a maximum 3.5 ng/m^ ■~gases) of (EPA al. 1985b, 1986, TCDD of in following accidental as estimated body body, 0.05 the daily ng. 50 pg/m^. failed of to of the the pg/m^, but the days (EPA of et fg/m^. of atmospheric concerning of the however, from its European incinerators, of the the and Ballschmiter of al. The Office was levels are emitted 1986, to 0.47 Marklund in containing dropped to a et al. detection of Rappe can the of New Following an 2 , 3 , 7 , 8 - T C D D __ air 15 p p m level release of in Binghamton, pg/m^. concentration 2,3,7,8-TCDD 1986, fires concentration Building 0.23 the Silvex concentration Smith gas the of 38 incinerators; conditions air. State fire, concentration 0.62 sources transformer/capacitor in in Sweden, application Using incinerators 2,3,7,8-TCDD flue in U.S. Nottrodt the accidental fire isomers. concentration the was could not e s t i m a t e d as distinguish in in both vapor Indiana, 1986). certain air of its The isomers of publications ashes 1.3 ng/m^; 1983, was the p r i m e (under n o r m a l i z e d the 7.2.2 one 2,3,7,8-TCDD after 1985b, the that municipal ambient stacks are 2,3,7,8-TCDD The low tetra five thè fly to locomotive so method used other the m a x i m u m and Hites the is in Bloomington, from number Barnes an air from in field surrounding a 2,3,7,8-TCDD was of 10 fg/m^ in 200 1985). Water No report is available water, using methods range; however, effluents, 2.3.7.8- in TCDD, from none and samples a mean residences 2.3.7.8- TCDD has from ranging in 2,3,7,8-TCDD detected in aqueous from hazardous from a (detection effluent limit, from Dow been reported a waste disposal level of from the from none sediments to b e site 14 ng/L. Love detected from storm 10 waste to drinking to into industrial sites. in manufacturing 30 pg/g) to sump area 1560 facility 100 pg/g. the Tittabawassee 15 pg/L. Jacksonville, sewers, The including approximately The Canal in the nanogram-per-liter trichlorophenol leachates 2,3,7,8-TCDD been leachates in effluents detected 2,3,7,8-TCDD and limits dibenzo-p-dioxins, in Mi c h i g a n , leachate the tetrachlorinated discharged wastewater River, on detection 2,3,7,8-TCDD has of ranged with sediments, concentrations of the human of consumption analytical methods. analytical a vast involving amounts 2.3.7.8York, of Eitzer Accidents larger had the ENVIRONMENT air gases around incinerators 2.3.7.8of The d i s p e r s i o n model, in of ambient fg/m^. concentration 2.3.7.8- TCDD The consumption from tetrachlorodibenzo-p-dioxin in identify 2,3,7,8-TCDD was the estimated ambient unequivocally measured to in capabilities 18 et the From in IN THE approximately most pg/kg. been ESTIMATED through resulting Air detection and 20 of of breast burden 7.2 consumption daily pump water in N e w Y o r k ng/L. The residential The Arkansas, from contained concentrations sump water, 69 and surface limit, water around 10 to 100 pg/g) Lamparski et al. 1986). 7.2.3 same 9570 site ng/g were (EPA none 1985b, detected Tieman (detection et al. 198 5 , Soil Concentrations below the soils, ng/g. of detection the level 2,3,7,8-TCDD limits of industrial accidental 2.3.7.8- TCDD in is apparent of still-bottom (2,4,5-TCP) 7.2.4 been sites, spillage It of of from residue the range disposal sites, and the m a n u f a c t u r e largest source of sites are accidental of to 0.009 originated The given or from involved 2,3,7,8-TCDD. locations the are In urban of <0.0002 that containing soils methods. samples that from the in different 7.1 uncontaminated analytical in chemicals soils may be is detected waste from Table in most current 2,3,7,8-TCDD 2,3,7,8-TCDD has certain in levels in T able improper of 7.1. disposal 2,4,5-trichlorophenol 2,3,7,8-TCDD in soils. Other There crop the to are plants limited data (Anonymous 2.3.7.8- TCDD (up few parts per trillion to roots_of these plants, than the surrounding crops would that 1985). 752 ppt) of grown after the 2,3,7,8-TCDD however, soil, demonstrate 2,3,7,8-TCDD Crops Seveso in the that contamination a not bioaccumulate accident contained levels similar of the in contaminated with aboveground contained higher suggesting the does in soil of a The 2,3,7,8-TCDD study edible only portions. using portion root of the plant. Since aquatic investigators pollution Huron in the contained collected Samples from from Fish of to 67 pg/g a l. 1984). from to contain and Great Michigan et Lakes and control the tissue 60; samples et organs: lung, is of fat, 60; a poor Lake 1986, pg/g 2,3,7,8-TCDD. maximum levels Superior Buser from Lake eggs and had Rappe were limit, Pond, Massachusetts, (Buser soybean, limit 1986, and Rappe 1984). crawfish samples and of limit 10 pg/g, of or minimum 1984). rivers (detection of the from Woods the chief 2 pg/g) Ryan et were No in from Canadian 2 to 4 pg/g contributor to the b o d y burden analyzed fat has a woman who kidney, 101 selected Michigan detection That 1840; gull ( R y a n et 1985). is following sample Niemann detection a gull 1985, investigators analysis The at al. tissue many a and few herring the amounts et 2,3,7,8-TCDD at to a of possible al. in rice, populations. Italy. blood pg/g 12 al. and (Fehringer samples found adipose TCDD, different 26 Firestone Since 2.3.7.8- that the Louisiana and pork 1985a, brain, contained (Stalling indicator herring Saginaw Bay had from Lake Yellow perch Arkansas Seveso, and 2,3,7,8-TCDD TCDD was al. Lakes those 2.3.7.8chicken Great Ontario an Similarly, contain between undetectable and found the 2,3,7,8-TCDD. 2,3,7,8-TCDD, as suspected water b o d i e s . A contamination samples shown bioconcentrate fish-eating birds 75 p g / g Lake 2 . 3 . 7 . 8 - TCDD; levels organisms analyzed levels pancreas, 40; indicator 7 months from both burden after is the 2,3,7,8-TCDD (pg/g) 1040; 150; and blood, of.the tissue the h i g h e s t died of fat liver, 6. This 2,3,7,8-TCDD confirmed by accident were in found thyroid, analysis body of exposed 85; suggests burden for in Table 7.1. Lerels of 2,3,7,8-TCDD in soil from different locations S ite Sam ple history T C D D concentration*4 (n g /g ) References Love C anal, N Y Soils outside the dum p site Jacksonville, A R W aste disposal site N D -2.9 EPA 1985b M idland, M I Inside D O W facility 0.01-52 N estrick et al. 1986 St. Louis, M O U rban sam ple o f no obvious source o f contam ination 0.12 EPA 1985b S henandoah S tables, M O C ontam inated by w aste oil 101-33,000 Tiernan et al. 1985, K im brough et al. 1977 T im bcrline S tables, M O C ontam inated by w aste oil 30-42 T iernan et al. 1985 Bliss F arm , M O C ontam inated by w aste oil 382* ' T iernan et al. 1985 Bubbling S prings R anch, M O C ontam inated by w aste oil 76-95 Tiernan et al. 1985 M inker R esident, M O C ontam inated by w aste oil 50' Tiernan et al. 1985 T im es Beach, M O C ontam inated by w aste oil 4.4-317 T iernan et al. 1985 U rban areas, U nited S tates U rban sam ples o f no obvious source o f contam ination N ew Jersey Spillage of 2,4,5-TC P still bottom 26,000* Jackson et al. 1986 N ew Jersey S crap yard where used reactor vessels were collected , 1,100* Jackson et al. 1986 Lansing, M I U rban sam ple N D (0.0007)-0.003 N estrick et al. 1986 G aylord, M S U rban sam ple N D (0.0002) N estrick et al. 1986 D etroit, M I U rban sam ple N D (0.001-0.020) <0.0002-0.009 0.0021-0.0036 EPA 1985b N estrick et al. 1986 N estrick et al. 1986 ì V Table 7.1 (continued) S ite Sam ple history T<£DD concentration® 4 ’; (n g /g ) R eferences 0.0042-0.0094 N estrick et al. 1986 C hicago, IL U rban sam ple A kron, O H U rban sam ple 0.0063 N estrick et al. 1986 N ashville, T N U rban sam ple 0.0008 N estrick et al. 1986 P ittsburgh, PA U rban sam ple 0.0026 N estrick et al. 1986 Philadelphia, PA U rban sam ple 0.0009 N estrick et al. 1986 Brooklyn, N Y U rban sam ple 0.0026 N estrick et al. 1986 A rlington, VA U rban sam ple N D (0.0003) N estrick et al. 1986 °N D = not detected. ^V alues w ithin parentheses are detection limits. rO nly one sam ple was analyzed. CD CD 3 I 72 humans. The population levels of of 2,3,7,8-TCDD the U n i t e d 20 p g /g, with Adipose Tissue Survey, 76 % . Europe, th e In tissue of a mean value population 3 pg/g. Instances in individuals For individuals in State the 2,3,7,8-TCDD mean value of individuals pg/g and (1.4 al. 37 p g / g ; (Patterson in of lightly 1986a; Nygren the bod y 1986). pg/g) and al. 1986). from to 3-3 pg/g, human et 2.3 pg/g, a mean obtained of 1987, Nygren al. This in near a mean the 1986, of in EPA of Other than however, for into human the tissues, detected for to as always obtained et pg/g; in (0.1 al. There towards al. two milk 6.0 pg/g) Seveso, South in also Vietnam,~ sprayed areas; 1.3 detected et a at (Heath a l . 1986, large concentrations groups mother's to in and Yugoslavia is 6.4 It has and Germany, Rappe in milk has been levels > 1 0 contaminant in of in found ng/g. any pentachlorophenol found Given in this samples from finding, a incinerators in 1986). the with muscle, 1986; to b e to areas; of n e w m u n i c i p a l from 2,3,7,8-TCDD given area; pg/g; been incinerators. (Nygren 2.5 a in commercial construction effect analyzed reported it h a s industrial waste put were none other 1985, et 2,3,7,8-TCDD 2,3,7,8-TCDD, 1986; median (Rappe from mothers analyzed by al. contributor control 1984, et in h u m a n b r e a s t accident 1985b). control respectively detected 0.6 of other reported are Netherlands, Young values mothers limits pg/g, and 2,3,7,8-TCDD. 2,3,7,8-TCDD was the to for 99 in parentheses) largest none 50 p g / g 1986, rarely been 1986); moratorium Sweden was liver, to pg/g. al. In addition product and a level al . The been determined and the 750 pg/g), the the et 1984). 1,2,3,7,8-pentachlorodibenzo-p-dioxin (Hagenmaier Canada, et difference congener has 1-year areas with South Vietnamese commercial municipal been from Denmark, Jensen contain 40 1.9 et investigators. / *0 to Patterson to to States, from mothers pg/g) 1986, from (2.8 detection United al . milk (the 2,4,5-T-exposed to unexplainable with exposed exposed, in breast-fed babies 2,3,7,8-TCDD have (0.5 with milk levels 28.0 p g / g trace 17 p g / g 2,3,7,8-TCDD countries: Italy, Sweden, of in Weerasinghe have is from 2.3 in M i s s o u r i analyzed in m o t h e r s N e w York, pg/g, 3 pg/g) Schecter Young been of detected 1986; 1986; milk different none 1984; also following parentheses) or exposed few heavily possibly that human breast intake The a 28.3 (concentration range 20.2 reported of to compared with al. al. al. populations et few in B i n g h a m t o n , 11.6 difference help et et et 2,3,7,8-TCDD Hu m a n breast milk has been Gross Stanley no exposed, sought medical 1987c; 1986; however, exposed to during capacitor to of (EPA control a a have 2,3,7,8-TCDD who concentrations tissue with tissue either of of adipose 9 pg/g, accidental tissue fire the adipose chemical range in in V i e t n a m h a d group et adipose to in during adipose the frequency limit, population al. this or Building in a to Human (detection Vietnam veterans et The levels general National concentration detectable to the operations found Patterson Office a U.S. detected with not the In in from undetectable spraying 2,3,7,8-TCDD was Graham is tissue ranged from undetectable a mean value in was 2,4,5-T concentrations involved 7 pg/g. of higher example, 17.4 pg/g. ranging to exposed containing fires. adipose Canada 2,3,7,8-TCDD general transformer with of of reported levels 5 in and 2,3,7,8-TCDD range spraying herbicides had of the mean value been States autopsy the none of two following detected; subjects results: and kidney, in 73 none detected. range in of 1 the b l o o d N e w York, The to 4 of at a detection pg/g (Ryan 2,3,7,8-TCDD Orange in V i e t n a m during Disease Control range with 2.3.7.8- limit levels to none detected pg/g. Serum of (0.0013 pg/g) 1970, a to same study 7.3 OCCUPATIONAL EXPOSURES however, Occupational containing 2,4,5-T. to p u r i f y The manufacture of evidence occupational veterans and Binghamton, 7.4 From use AT of the data that m a y and in (SRI States industrial near municipal at sites of are or population improperly exposure in near controlled expected Studies in humans Sects. 4.3.3 2.3.7.8- TCDD results Animal infants in is may and a than during the of between 3.9 in the the during 1984). is the in heavily State the these step that products pur i f i e d products. (Rappe Office transformer of the The indirect significantly exposed Vietnam Building in fire. than are the may municipal to have not studies, represent 4.3.4 on flue are and risk. ashes exposure. exposed or other that sensitive fetal and in the residing through suggest thé general to near the is a sensitive fetus As reproductive and mice, in monkeys. 4995 of milk. population. death Workers sources there that rats in residing trichlorophenol, and mothers from workers Populations to (from and manufactured industries) teratogen and gases or to 2,4,5-T longer potentially developmental abortions exposure 2,3,7,8-TCDD, 2,3,7,8-TCDD a no subjected disposal to population. are contain demonstrated demonstrated spontaneous 2,4,5-T nursed by such 2,3,7,8production general however, of is and incinerators receive of hexachlorophene, at h i g h e r sites it and the associated those levels susceptible also be 7.3, in Since b o t h to b e and involved however, incinerators 7.2 population to h i g h e r workers salts, 1987). Sects. general salts, 2,4,5-T and in the exposed Breast-fed babies are newborn the chemical waste 2,4,5-T, residing subpopulation. serum 2,3,7,8-TCDD during since 2,3,7,8-TCDD discussed its incinerators 2 . 3 . 7 . 8 - TCDD. occur occur compound the its expected improper hexachlorophene, to for 25 range 30-fold higher 2,3,7,8-TCDD groups, 2,3,7,8-TCDD 2,4,5-trichlorophenol profession to to at be higher this may available the segments containing and are of other herbicides the U n i t e d and a median value to b e 2,3,7,8-TCDD following trichlorophenol municipal reported contaminants, exposure occupational of study to to A g e n t Center HIGH RISK groups levels exposure its exposure the m o n i t o r i n g the reported the pg/g) exposed the 1985). (0.0013 2,3,7,8-TCDD may certain workers to p r e d i c t Among of levels N e w York, occupational TCDD. with al. exposed reported by same in detected Binghamton, et heavily detected The in (Schecter were been 12 pg/g, chemicals tissue POPULATIONS possible or these in have pg/g. been to occupational adipose who occupationally from levels the higher accident 2 pg/g of non-Vietnam veterans heaviest 2,4,5-T on of to was of hexachlorophene, trichlorophenol, and herbicides contain much higher data have exposures and use No 1 were 2,3,7,8-TCDD 1987). production is u s e d 3.8 and prior (MMWR, the group group determinations following of bet w e e n none in a in these No 1967-1968 levels levels in 1985b). in veterans a median value TCDD al. exposed workers detection Serum pg/g, limit et and discussed toxicity, and also Since these 74 effects occur doses that that, at certain sensitive low route stages can be for doses, appear subgroup. 2,3,7,8-TCDD major at do n o t receive indicate in the adversely fetal data ingested elimination newborn might to of Animal were found and to case greater mother, the demonstrate and 2,3,7,8-TCDD. exposure that newborns teratologic the development, also during nursing of of affect than that This the are more fetus that effects it is at likely represents toxic lactation indicates adult, since sensitive than a levels is of a that the no data adults. 8. Several different and 8.2. methods media. The methods illustrative samples are Some of before a ANALYTICAL METHODS available of the listed few more in recent for the analysis recent methods these tables methods. are not analysis are important, 2.3.7.8- TCDD in m o s t samples are low. stack that exist both in details of (1986), and The accuracy availability use as an of of internal combination of analysis of gas has several methods (MS) 2,3,7,8-TCDD sensitivity to all (EI/MS), al. The 1985). because these HC1, permit solvent Best access The HC1 to the transfer from et of analysis channel may be Jensen adipose indicators of fish catfish yet crystal Ozvacic a were 1986). carp) the of surface has milk, of since fish and hot water fly ash the ash to on the two 2,3,7,8-TCDD. (Ryan (e.g., bottom et tissues The feeders feed upon to improve fly these in excess to placed some bot t o m the extraction removes 2,3,7,8-TCDD-polluted water in present of the et challenge structure of electron (Buser digestion with because that than difficulty been for levels. a higher congeners residue pore Emphasis those and low special using the the of freeze-drying important for a 2,3,7,8-TCDD and number and mother's and for recovery obtained by and the cleanup, dibenzo-p-dioxins a opens sample shows poses large with With at v e r y (NCI/MS) the h u m a n b o d y b u r d e n also the TCDD. commonly HRGC Fourier used analytical untested methods Some capillary isolation More for The 2,3,7,8-TCDD identification samples the hydrophilic al . is generally 2.3.7.8- and (1986), of a l. feeders 1984a, 1987). Besides but from tissue of ash for performed freeze-drying and indicators phase. years l^C) analysis. sensitivity extraction solvent, (Stieglitz be fly treatment solvent analysis can be lower results material may a of followed by toluene. important (HRGC), and high-resolution polychlorinated it h a s and available (GC) ionization MS 2,3,7,8-TCDD samples. dilute with of other analysis of poor resolution 8. 1 a re collecting in V e l z y in recent (^Cl (HRMS), unequivocal chemical impact MS available but concentrations and particulate spectral chromatography quantification negative for the is p a r t i c u l a r l y increased in mass mass^spectrometry of are isotope-labeled standard one exhaustive since in in T a b l e s (1986). stable high-performance Although the v a p o r stack-sampling methods Brenner This 2,3,7,8-TCDD given Methodologies their samples of are of these column methods (Naikwadi transform can be found (1984), and descriptions in B u s e r EPA et of al. are GC with (1985), Tieman newly GC of liquid with matrix (Wurrey et al . 1986), (Mohamad et al. 1986). al. for 2,3,7,8-TCDD (1985), (1985b). 75 developed analysis a polymeric methods et the 1986), detection analytical other for spectrometry plasma the methods, available and Karasek infrared with microwave-induced detailed are 4997 Rappe 76 8.1 8.1.1 ENVIRONMENTAL MEDIA Air, See 8.2 8.2.1 Water, Table Soil, and Food 8.1. BIOMEDICAL SAMPLES Fluids/Exudates See Table 8.2. and Tissues ) V T ab ic 8.1. A nalytical methods for environmental samples Sam ple matrix Sam ple preparation Ambient room air Vapor phase sample collected following accidental by silica gel; particulate transformer Tire sample collected on glass fiber 1 1 : Detection limit A n alytical method0 H R G C /H R M S ' 0.003 pg/m ’ A ccu racy0 References 131 ± 27% at Sm ith et al. 1986 5-10 pg/m 1 filter, solvent extracted, and precleaned by alumina and carbon J Ambient outdoor air Vapor and particulate collected H R G C / N IC I/ M S on polyurethane foam and glass fiber, precleaned by Florisil 0.1-0.2 pg/m 5 (method detection limit too high NA Oehm e et al. 1986 NA Eitzer and for T C D D determination) and modified silica V apor and particle collected on H R G C / N IE C / M S -S IM polyurethane and glass fiber, precleaned by silica and alumina S la c k emission Vapor and particle collected on N R ( T C D D not separated from other tetra isomers) H R O C /M S NR H ites 1986 NR X A D - 2 and glass fiber by modified E P A method 5; solvent extracted H agenm aier el al. 1986 -v j and precleaned by silica, alumina, and Biobead F ly ash from municipal incinerator Solvent extraction, H P L C separa­ G C / M S or H R G C / F I D tion on normal-phase and reverse- N R ( T C D D not separated NR Tong et al. 1984, Tong and NR Lam parski and from other tetra isomers) Karasek 1986 phase column Solvent extraction, precleaning G C /LR M S 40 pg N cstrick 1980 by two adsorbent columns, and further fractionation on reverse- CD and normal-phase H P L C CO CO Solvent extraction, precleaned on H R G C /LR M S NR NR Buser and 3 ng/g NR Donnelly Rappc 1980 silica and alumina Soil Solvent extraction, K O H wash, pre­ H R G C /LR M S et al. 1986 cleaned in alum ina, reversed- 0O lS'd phase H P L C , and carbon Solvent extraction, K O H wash, pre­ cleaned in chem ically treated silica, basic alumina H R G C /LR M S NR NR Freeman et al. 1986 Table 8.1 (continue^) Sam ple matrix W ater Sam ple preparation Sam ple passed through glass fiber A n alytical method“ G C /M S filter and adsorbent cartridge; solvent extracted and precleaned by acid alum ina, graphitized carbon and alumina C hicken and pork Fat and liver solvent extracted, partitioned with concentrated H R G C /M S/M S l l , S 0 4, cleaned by Florisil and reverse-phase H P L C Fish Homogenized fillet digested with ethanolic K O H and solvent H R G C /M S extracted; extract cleaned by silica gel-supported H 2S 0 4 column and H P L C Homogenized fillet solvent H R G C /H R M S extracted, partitioned with con­ centrated H jS 0 4, cleaned by Florisil Homogenate digested with concen­ H R G C /M S/M S trated H C I and solvent extracted; extract cleaned by silica gelsupported H jS 0 4, chem ically treated silica and alum ina, and reverse-phase H P L C Digested with lalkali, solvent H R G C /EC extracted, washed with concen­ trated H ] S 0 4, and cleaned up by site exclusion chromatography, normal- and reverse-phase H P L C Fish and herring gull Solvent extracted, cleaned up by potassium silicate-silica gel. cesium silicate-silica gel. carbon, and H , S 0 4-silica gel cesium silicate and alumina H R G C /LR M S Detection limit !!| p g / L 2-4 pg/g Accuracy' References 88% at O 'Keefe 6.5 p g / L et al. 1986 NR Ryan ct al. 1985a 5-10 pg/g NR Fehringer el al. 1985 2-10 pg/g NR Ryan et al. 1984 <1 pg NR Clem ent et al. 1986 12 pg/g 105% at 18-45 pg/g Niemann 1986 1-8 pg/g NR Stalling et al. 1983, Rappe 1984 vj 00 r r \ Table 8.1 (continued) Sam ple preparation Sam ple matrix A n alytical method" \ Detection limit A ccu racy* NR 72% References ' . Fish, egg, or sediment Added M CI and solvent extracted; H R G C /EC /LR M S extract cleaned by gel permeation, Law rence et al. 1986 trisodium phosphate, H ] S 0 4, alum ina and carbon columns *H R G C ” High-resolution gas chromatography; N I C I / M S — negative ion chem ical ionization mass spectrometer; N I E C / M S — negative ion electron impact mass spectrometer; S I M “ selective ion monitoring; F I D — flame ionization detector; L R M S - low-resolution mass spectrometer; E C ■* electron capture detector; N R ■* not reported; N A — not applicable. vcj cn o o * /3 o V o _ Table 8.2. A nalytical methods1for biomedical samples Detection limit Sam ple m atrix Sam ple preparation A n alytical method“ 85% at 24 pg/g Lap cza et al. 1986, Patterson ct al. 1987a traction. cleanup by carbon and i silicate/silica gel Blood, liver, kidney Homogenized sample solvent extracted, and muscle cleaned up with H jS O ,, chromatography, IIR G C /M S /M S 14 NR R yan et al. I985b,c or Florisil Serum Isotopic Solvent extraction, concentrated sulfuric acid wash and cleanup by dilution with carbon and silicate/silica gel, followed H R G C /H R M S 1.25 X I0 '5 89% at 5.0 jig / m L Patterson et al. 1987b by s ilic a ( e / H ,S 0 4/silica gel and alum ina columns ° H R G C — High-resolution gas chromatography; N I C I / M S “ negative ion chem ical ionization mass spectrometer; H R M S — high-resolution mass spec­ trometry; M S — mass spectrometry; L R M S — low-resolution mass spectrometer; C I M S — chem ical ionization mass spectrometry; N R “ not reported. * cn o o \ o Oi-'o - \< A 9. 9.1 INTERNATIONAL No World Organization 9.2 REGULATORY (WORLD HEALTH Health AND STATUS ORGANIZATION) Organization advisory ADVISORY standards developed by IARC were found. The World Health which places is p r e s e n t e d b e l o w . NATIONAL 9.2.1 Regulations The reportable 2,3,7,8-TCDD CERCLA hazardous Water Act 9.2.2^ were substance ( F R 50, Advisory *^Air. quantity i n c a t e g o r y X. No. 65, (R Q ) The is p. for 2,3,7,8-TCDD authority provided for is 1 lb , listing by Section for levels 2,3,7,8-TCDD 307(a) of the as a Clean 13456). Guidance No health advisories (HAs) of 2,3,7,8-TCDD in air encountered. Water. AGENCY EPA ADVISORY Drinking water advisories: 1-day HA--1.0 x 10'^ (child) mg/L (child) Long-term HA--1.0 x 10*” mg/L (child) L o n g - t e r m H A - -3.5 X 10*® (adult) Drinking water 10"^ to mg/L EPA 1 0 ’6 m g / L X 10-day HA--1.0 Ambient 10*^ 10'^ (EPA equivalent excess cancer l e v e l - -3.5 x 10*® mg/L risk--2.2 x 10*® to 2.2 x 1 0 ‘H x 10*^ to 1.3 x 10*^2 1986b) water to mg/L quality 1 0 ”^ e x c e s s criteria cancer risk--1.3 m g/L ( E P A 1984) Food. AGENCY ADVISORY FDA Levels 9.2.3 Data by doses 2,3,7,8-TCDD Murray this et study, fish: No serious health concerns--<25 EPA (1985a) calculated data from ppt / (EPA 1985b) Analysis Reference for in al. rats (RfDs). based on (1979), were the as a reanalyzed by exposed to diets 83 a chronic three-generation Nisbet containing and Paxton in (1982). 2,3,7,8-TCDD 5005 oral study at RfD rats In levels p - 5 |0fc 84 that provided resulted effects on resulted in the LOAEL, doses of in d e c r e a s e d litter 0.001, fetal size in d i l a t e d and 0.01, and survival; fetal and renal pelvises, gestational index. Therefore, and was the RfD - (0.001 RfD where: calculated j j g/kg/day. middle neonatal the as The dose fetal dose of highest resulted survival. decreased The lowest weight, 0.001 dose in and dose changes pg/kg/day is a follows: / i g / k g / d a y ) / ( 1 0 0 ) (1 0 ) 0.001 fi g / k g / d a y - LOAEL, 100 uncertainty factor - 0.1 the - for 0.000001 inter- pg/kg/day and , intraspecies extrapolation, 10 - Carcinogenic in CAG when Group B2 uncertainty potency, when 2,3,7,8-TCDD factor q^*. EPA 2,3,7,8-TCDD is considered and/or chlorophenols. Group B2 humans is there are to inadequate, consider that there data to 2,3,7,8-TCDD are not support phenoxyherbicides (1982) has the_CAG only the 2,3,7,8-TCDD be exposure limited be EPA based on (1985a) the histologic evidence calculations lungs, diets were liver, different an based produced of (FDA) have Disease Kimbrough al. to et Squire an al . (1978a,b) on final (CDC) a virtual excess study, of cancer The FDA whereas the respectively, are 6.4, by dose of CDC to that and that of the the EPA. The tumors of the maintained different and , x hence, 105 (mg/kg/day) agencies. Both 2,3,7,8-TCDD, were calculations al. (1978a,b) derived by and 57.2 fg/kg/day. the Drug Administration (Hiremath dose on pathologic evaluation. and for 10"^ estimate for rats 1.56 other Food risk of in safe 27.6, et IARC analogous Squire two derived, safe is reexamination incidence tumor risk the a in the limited human in c o n j u n c t i o n w i t h cancer and The used data indicates carcinogen 2 years. and in carcinogen. incidence calculations Kociba a virtual unit each pathologic methods for et which al . based on were based study. EPA, 1986, the Thus, CDC, Kociba on the human and FDA, STATE Minnesota, on for B1 feasible. turbinates value Control intake values 9.3 for potency, 1984). evaluation increased Bl recommended conducted by and nasal Group which occupational (1978a,b) study the 2B, evidence also a human (1984) extent al. differences The calculated corresponds that the values Carcinogenic for et 2,3,7,8-TCDD q^* values. average Center from B 2 . NIOSH a quantitative Kociba h a r d palate, containing examinations was developed study by data but is 2,3,7,8-TCDD in Group carcinogenicity 2,3,7,8-TCDD, in Group fullest although animal animal that and phenoxyherbicides carcinogen. a potential the classified alone, human chlorophenols, of Group to that sufficient sufficient considered a LOAEL. has considered indicates 2,3,7,8-TCDD classification of in a s s o c i a t i o n w i t h a probable and/or (1985b) is consideration classified for use the either allowable have Michigan, of 2,3,7,8-TCDD. regulations or are r e g u l a t i o n s . (Regulations still being and N e w Y o r k have levels compiled at and the in the advisory time In set additional addition, process regulations states may of promulgating guidance of printing.) other from the states were et 10. Abernathy DJ, Gre e n l e e WF, H ü b a r d JC , Tetrachlorodibenzo-p-dioxin C3H/10T1/2 Adams WJ, cells. Blaine 2 , 3 , 7 , 8 -TCDD. Adams WJ , K.M. 1986. 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In: Complex Scientific Chromosome L. Banki, Task. examinations ed. The Medicina: in w o r k e r s Development Budapest. of a 239-256. producing Pesticide (Cited in EPA 1985a) - Czuczwa JM, Hites polychlorinated Czuczwa and JM, fates. *DeCaprio 1986. the guinea L, receptor as Mol an an L, of des 27: and PE . O'Keefe of with 1981. the Technol; and 18(6): 444-450. diben z o f u r a n s : Sources PW , Rej R, S i l k w o r t h JB, Kaminsky 2,3,7,8-tetrachlorodibenzo-p-dioxin a PCB-containing transformer fluid presence the LS. in 6:454-463. tissues and Carcinog Mutag; D'Argy combustion-generated 195-200. Association between murine for Sci dioxins 20(2): of R, Aim to of Ah T C D D -i n d u c e d 1:399-406. G. toxicity the sensitivity 1985. of Fetal thymus 2,3,7,8-TCDD organ and its culture congeners. 133-140. New Zealand. into defects Rosiers DM , fate Environ Airborne Technol; Toxicol; E, model Health, Z e a l a n d M e d J; furans. toxicity Teratog Investigation PCDDs RM . Hassoun congenital and Comparisons Pratt Pharmacol; Dept, oral in embryonic in vitro Environmental 1986. Sci Fund Appl palate. Dencker RA. McMartin pig: pyrolysate. cleft Hites Environ AP, 1984. dioxins Subchronic Dencker R A. 1980. allegations and 2,4,5-T 314-315. (Cited 1986. of an spraying in EPA Methodologies related compounds. Report to the M i n i s t e r of Health of association between human in and around teKuiti. 1985a) for materials Chemosphere; 15 : New ' contaminated with 1513-1528. C, O' o io 89 DiLemia R, Crimaudo effects on satellite application (Cited C, in environmental in EPA D o n n e l l y JR, Vonnahme 8280 C h o u d h a r y G, Keith Perspective. Chelsea, for Eitzer BD , in atmosphere. EPA. Hites 1979. 2,4,5-T EPA. TL, LH , Hedin eds. MI: of EPA. 1985b. Health Criteria (PCDD) and EPA. EPA. of Assessment No. 1987. (CDDs mixtures Hardell and L, dibenzofurans six-year in relation (Cited Document for in EPA for to forest 1985a) 2,3,7,8- 2,3,7,8-Tetrachlorodi- for Polychlorinated Office of Health the FY82 National Polychlorinated EPA, Criteria Dibenzo-p- and Environmental for (PCDF). EPA Report DC. estimating chlorinated Human Adipose Dibenzoz-p-dioxins Washington, Washington, Health October risks 625/3-87/012. J, SPW , No. 560/5- D C. a d v i s o r y , -------- 16, 1986. Draft. associated with dibenzo-p-dioxins R e p o r t No . McClain L, B e r g NO, and dibenzofurans Risk Assessment 27-33. in T, of 76: Forum. Vietnam Veterans: JAMA; 252(7)'.903-912. 1979. soft-tissue 3872-3875. Axelson substances: EPA 1984. A x e l s o n 0. the M o l l e r T, chemical (Cited al . defects. Moller tumors O ' B e r g N, to et with birth Lakartidningen; exposure 38: and 215-230. OH. Dibenzofurans mesenchymal substances. Ind Med; of IV. EPA, babies Hardell E r i k s s o n M, J Mulinare on malignant sarcomas areas in 399-435. DC. fathering E r i k s s o n M, chemical Voi. Substances, of a n d C D F s ) . EPA. for dioxins EPA. of C, DC. Interim procedures to E r i c k s o n JD, study of Toxic pp . 2,3,7,8-Tetrachlorodibenzo-p-dioxins. EPA Washington, risks Document Scan Analysis Polychlorinated Evaluation 600/X-84-194-I. Environmental 600/8-84-014, D r i n k i n g Water, exposures 42-47. and Dibenzofurans 27(3): Criteria Document EPA Cincinnati, Specimens. Office 1986b. 61(1): EPA-440/5-84-007. No . Broad Survey Office and TCDD for investigation of Oregon Quality EPA. 86-038, model dibenzofurans. Rappe of a field Subst, EPA Washington, 1986a. Genet; Inc.; Chem; Off Toxic Assessment, Tissue Publishers, in three EPA Report EPA Report and Concentrations Drinking Water a nd .Assessment Office, Dioxins. 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Poncelet Mutagenic Hileman Chlorocarbons 15 : S, and F D, Bioconcentration Chemosphere; Mercier, M. 1980. (Cited EPA 2,3,7,8- genotoxic effect of Mutat Res; W e n d l i n g JM, tissue from Genetic in a review. (2,3,7,8-TCDD) effects of Arch 1985a) 168:241-242. W i l s o n JW. a Missouri in 15:1495-1502. S a lm o n e lla ty p h im u r iu m . on 533-541. O r t h RG, adipose Moreland FS. Toxicol Appl rat. Green S, Moreland, 1975. FDA By-Lines; Sheu Greenlee WF, mechanisms (Biol. Lech. Greenlee action of WF, of 6: C. 292. Osborne Cytogenetic Pharmacol; R, toxicity 33: 1977. tetrachlorodibenzo-p-dioxin DC. 1986. 1986. population. Chemosphere; 1595-1600. Green the in F, azobenzenes 9(5): tetrachlorodibenzo-p-dioxin, G r a h a m M, F. including humans. Contam Toxicol; AK. Korte 2,3,7,8-tetrachlorodibenzo-p-dioxin organisms chlorinated Environ of on (Cited Cytogenic rat bone (Cited in E P A H u d s o n LG, of TCDD evaluation 161. several EPA of dioxins in 1985a) 2,3,7,8- cells. FDA, Washington, 1985a) Toxcano to h u m a n effect marrow of in WA Jr. 1984. epidermis. Studies Banbury Rep; on the 18 D i o x i n A c t i o n ) : 365-372. Dold KM, 2,3,7,8-TCDD Irons on RD, Osborne thymic R. 1985. epithelium. Evidence Toxicol Appl for direct Pharmacol; 79: 112 - 120 . G r e i g JB. and its 1979.-The structural toxicology analogues. of 2,3,7,8-tetrachlorodibenzo-p-dioxin A n n Occup; 22:411-420. (Cited in toxicity of EPA 1985a)* * G r e i g J. 1984. Differences between 2,3,7,8tetrachlorodibenzo-p-dioxin D i o x i n A c t i o n ) : 391-397. skin and in mice. liver Banbury Rep; 18 (Biol Mech 92 *Gross ML, L a y Jr, JO, Lyon PA, Tetrachlorodibenzo-p-dioxin Veterans. E n v i r o n Res; Hagenmaier H. 1986. and Hagenmaier H, K r a f t M, Comparison of various H a n i f y JA, 15: al. 1984. of related products. J a g e r W, 2,3,7,8-TCDD M a y e r U, sampling methods P, Nobbs C L, W o r s l e y RJ. Science; 212:349-351. L, 711-717. Hardell to (Cited S t a n d s t r o m A. 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Health effects 2,3,7,8-tetrachlorodibenzo-p-dioxini JAMA; of 255: 2031-2038. Holden 77-772. / Vi* C. 1979. (Cited Agent Orange in 1985a) EPA furor continues to build. Science; 205: 93 Holmstedt, Toxicol; HSDB 1980. Huetter under EPA 1982. conditions. S, Ehrenberg on bacterial L, 1987. 6, I 18. Arch National Library of 1987. Studies on microbial Pergamon Lofroth systems. Hutzinger 0, PCDDs and PCDFs: B l u m i c h MJ, IARC (Intemation Agency An Chemicals Ser metabolism Environ Gejvail 1: T. 32-33. V a n d e r b e r g M, overview. to H u m a n s . G, Ambio; Sci; on Lyons, the 1972. 5: of TCDD 87-93. Olie K. (Cited EPA effects of 1985a) Sources and fate of 581-600. on Cancer). WHO; in 1985. 14: 1982. Evaluation of France: Mutagenic (Cited Chemosphere; for Research Organization-1ARC monographs of Bank). April Ecclesiastes 1985a) 1985a) Hussain TCDD M. Seveso in EPA Data print-out. Phillippi laboratory to (Cited Substances On-line R, Prolegomena 211-230. (Hazardous Medicine. in B. 44(4): IARC the World Health Carcinogenic V o l . 1-29, Risk Suppl. 4; 238-243. Ideo G., B e l l a t i G, Bellobuono 1 9 8 2 ^-"Increased u r i n a r y an area polluted with C h i m i c a Acta; Jackson DR , Solubility G, A, Mocarelli D-glucaric acid tetrachlorodibenzoparadioxin Roulier M H . - G r o t t e n HM, 2 , 3 , 7 , 8 -TCDD in Rust LH, eds. Chlorinated Dioxins Chilsea, MI: Lewis Publishers, Inc.; 1987. and polychlorodibenzofurans adipose tissue. Johnson FE, Kugler Total MA, Environ; Brown chlorinated phenols. Lancet; Jones 1986. C h e l s k y M. cutanea tarda Kaminsky LS, The of environmental role chlorinated A, Brambilla children P. living in (TCDD). Clinica and TCDD DeCaprio SM. AP, dibenzodioxin 64: G i e r t h y JF, and in C, Choudlary Perspective. in h u m a n milk, and blood and 259-93. Soft-tissue 40 . (Cited discussion Arch matrices 1986. Rappe (PCBs), polychlorodibenzo-p-dioxins 1981. Further JS. In: 185-200. (PCDFs) 2(8236): exposure. Warner, soils. and Dibenzofurans pp. Polychlorobiphenyls (PCDDs) Sci SW, contaminated Keith RE, Marocchi 120:273-283. of J e n s e n AA. P, excretion by sarcomas in EPA concerning porphyria Environ Health; Silkworth JB, experimental dibenzofuran and 1985a) toxicity. 41(12):100-103. Tumasonis C. vehicles in 1985. Chemosphere; 14:685-695. Kaye Cl, Rao chromosomal J S, Craniofacial Kellin CK, Simpson damage Genetics Christian induced weight loss SJ, in males BJ, Rosenthal exposed FS, and Developmental Inhorn in mice, tetrachlorodibenzo-p-dioxin. SL, C o h e n MM. to A g e n t Orange Biology P e t e r s o n RE. rats'and Fund Appl guinea and Evaluation their Supplement; 1985. pigs Toxicol; 1985/ of families. 1:259-265. Hypophagia- treated 5:700-712. with 2,3,7,8- 5015 i 94 Khera K S , Ruddick effects and the 70-84. (Cited in K i m b r o u g h RD, pathology Health; of 32: JA. EPA Polychlorodibenzo-p-dioxins: Perinatal lethal test C D, Liddle JA, Kimmig F a l k H, Toxicol SJ, Stehr P, Fries (Ger.) K n u t s e n AP. in EPA 1957. 1984. RJ, dioxin (TCDD). *Kociba RJ*, chronic toxicity p-dioxin in rats. *Kociba BA. akna B e y e r JE, Toxicol DG, Appl et Environ Keyes Sei); DG, Ann 1986. and PCDF of al. I V, JC, Campbell Harris on Lamparski heptaand 1986. 281-287. (Cited of chlorakne) 115: durch 540-546. in humans. BS, Sei; Chirtel in E P A Results of 1985a) a 1978b. (Cited Toxicologic in rats. in EPA Gehring 320:397-404. and Ekarohita the rat. J PJ. 1979. M c K i n n e y JD, Birnbaum the induction of cleft palate (TCDD) (Cited LS. Med Long-term (TCDD) in EPA of in 1985a) different 15:2011-2022. N, Exp MW, Occup 1985a) Ezekiel distribution of Pharm studies Toxicol embryotoxicity Chemosphere; SJ, two-year 4 6 ( 2 ) :279-303. absorption and in Bull 2,3,7,8-tetrachlorodibenzo- C a r r e o n RM, M. 1986. 2,3,7,8- Therap; 1986. by in C57BL/6N mice. LL, N e s t r i c k TJ. 1980. Determination octochlorodibenzo-p-dioxin range. L L, Water Anal Chem; N e s t r i c k TJ, Perspectives dioxins: of residential 239:673-677. Effects of 2,3,7,8Toxicol Appl 84:115-124. trillion Lamparski of 2,3,7,8-tetrachlorodibenzo-p- (TCDD) on distribution tetrachlorodibenzop-dioxin Pharmacol; and Environ implications exposure 1978a. study and marmosets. the m e c h a n i s m o f thyroid hormones (sog. C a r r e o n RM. NY Acad tetrachlorodibenzo-p-dioxin Lamb Arch 2,3,7,8-tetrachlorodibenzo-p-dioxin Studies in mice L a k s h m a n a n MR, 4: B e y e r JE, animals. on Epidemiology episode. Health of TCDD Pharmacol; B e y e r .JE, laboratory Studies 1977. Dermatologie; 13:387-406. oncogenicity of R. 12C: ■- Toxicity Rev; DG, studies per effects 33:673-681. 1982. toxicologic PCDD RE. 1984. ather. and Keyes (De T o x i c o l Krowke Ser . 1984).' Drug Metab Keyes RJ , RJ, Chem 14:47-93. 2,3,7,8-tetrachlorodibenzo-p-dioxin Kociba Adv contamination Berufliche Immunologic Schwetz G. (TCDD) zyklische Contam Toxicol; Kociba of KH. aromatische (Cited Environ Environ Health; Schulz chloriette Cline tetrachlorodibenzodioxin poisoning 2,3,7,8-tetrachlorodibenzodioxin J rats. 77-86. K i m b r o u g h RD, soil. in W i s t a r 1985a) Carter a 1973. dominant of a Analysis. 52 : Frawley large scale isomers of tetra-, 'hexa-, in particle samples at parts 2045-2054. NN, Hummel RA, environmental Chemosphere; 15: Kocher study 1445-1452. CW, for et al. chlorinated 95 Lapeza for CR, the Jr., Patterson extraction in human adipose L a n g h o r s t ML, in human m i l k J, DG, tissue. of Anal tetra-, samples. Onuska Determination of Liddle of Chem; 1980. in 58(4): hepta-, Chem; W i l i n s o n R, dioxins JA. 1986. fish Automated apparatus 2,3,7,8-tetrachlorodibenzo-p-dioxin 713-716. Determination hexa-, Anal. F, Jr., enrichment S h a d o f f LA . concentrations Lawrence and 5 2: and 2037-2044. A f g a n BK. and of parts-per-trillion octachlorodibenzo-p-dioxins 1986. sediment. Methods in research: Chemosphere; 15 : 1085- 1090. Loprieno M, cytogenic Sbrana studies I, Rusciano on mice tetrachlorodibenzo-p-dioxin Chlorinated N e w York: Lu CH, Dioxins Baggs Toxicity RB, and Press; Redmond evidence Lascialfari exposed ( T C D D ) . In : and Related Pergamon D, and rats D, Henary EC , tetrachlorodibenzo-p-dioxin-treated nutrition. *Lucier , Toxicol GW, Ingestion (TCDD) Appl Rumbaugh of soil alters Pharmacol; RC , McCoy Z, enzyme T. 1982. In v i v o in EPA 0., et on the al., eds . Environment, 1985a) S c h e c t e r A, Gasiewicz alterations in guinea pigs fed by T A. 1986. 2,3,7,8total parenteral 84:439-453. Hass contaminated with hepatic Impact (Cited for metabolic Lari 2,3,7,8- Hutzinger Compounds: 419-428. D, to R, Harvan D, Albro P. 1986. 2,3,7,8-tetrachlorodibenzo-p-dioxin activities in rats. Fund Appl Toxicol; 6:364-371. L u n d g r e n K, rats Andries results exchanges by halogenated Luster MI, rodents in M, T h o m p s o n C, increased a-naphthoflavone: An aromatics. D e a n JH, Toxicol GW. induction animal Appl. B o o r m a n GA. treated with Lucier in v i t r o 1986. of model Altered immune myristate-13-acetate, and benzo[a]pyrene. Banbury Environmental Factors in H u m a n G r o w t h Lynge A E. 1985. manufacture Manara L, Coccia laboratory follow-up P, rodents Croci by 22:815-818. Marklund Kjeller S, Doughterty samples of cancer R. L0 , 1986. and pyrolytic Chlorinated Dioxins Publishers, Inc; pp. T. exposure functions H a n s s o n M, products. Vol 11, pp. 199-213. among workers incidence Br J In: of or of PCDDs Rappe and Dibenzofurans in C, Cancer; TCDD cholic T y s k l i n d M, Determination in phorbol-12- Rep, Prevention charcoal to Development; in Denmark. 1984. addition of Che m Toxic; H, study of phenoxy herbicides of 85:189-195. 2,3,7,8-tetrachlorodibenzo-p-dioxin, and treatment chromatid for h u m a n Pharmacol; 1982. Dioxin sister Rappe and C, Choudhary in 52:259-270. toxicity acids PCDFs Perspective. Iss to Ryan in chow. C, Fd Collazo inn i n c i n e r a t i o n G, Keith Chelsea, LH, MI: 79-92. 5017 eds. Lewis 96 Marklund S, Rappe, polychlorinated C, Tysklind, dibenzofurans leaded gasoline. Marple L, Chemosphere B r u n c k R, Throop, Berridge, L. L, octanol partition coefficient Environ Sci Safe Throop. 20(4): 16(1): Water Sci Identification solubility of run on Measurement of 2,3,7,8180-182. the water- 2,3,7,8-tetrachlorodibenzo-p-dioxin. 397-399. 2,3,7,8-tetrachlorodibenzo-p-dioxin metabolites Synthesis, biologic and toxic effects in the of the rat. 41:153-159. Safe S. 1986b. Synthesis, biologic 2,3,7,8-tetrachlorodibenzo-p-dioxin. G. of 20(2): major May cars 29-36. Technol; 1986b. from G, M a s o n G, 1986a. of 1987. in e x hausts Mason Toxicology; S. 1986a. Environ Marple, Technol; L. KE . dioxins (England); tetrachlorodibenzo-p-dioxin. B. Egeback and 1973. Chloracne from tetrachlorodibenzodioxin. the Br J and toxic Chemosphere; accidental Ind Med; production 30(3): properties of 15:2081-2083. of 276-283. (Cited in EPA 1985a) M c C a n n J.--1978. .. M c C o n n e l l EE . in various McConnell Unpublished 1985. species E E, study. Comparative of animals. M oore JA, a single oral dose. toxicity in EPA of DW. 1978. in rhesus Toxicol Appl 1985a) PCBs Environ Health Dalgard tetrachlorodibenzo-p-dioxin (Cited and related Pers; Toxicity monkeys Pharmacol; 60: of compounds 29-33. 2,3,7,8- (Macaca m u l a c e a ) 43:175-187. (Cited following in EPA 1985a) ♦McConnell EE, Lucier Bioavailability GW , after Rumbaugh ingestion by RC, et rats al. and 1984. Dioxin guinea pigs. in soil: Science; 223:1077-1079. M c N u l t y W. (TCDD) 1984. Fetotoxicity for Rhesus Macaques M c N u l t y W. 1985. in Rhesus Macaques McQueen birth EG , Veale defects. Toxicity AMO, in EPA 1985a) A l l i s o n DC , Res; doses m ila tta ) . Am fetotoxicity Alexander of Meyne Mutat and Report (Cited J, 2,3,7,8-tetrachlorodibenzo-p-dioxin J Primatol; of TCDD, (Macaca m u l a t t a ) . E n v i r o n H e a l t h Health. Hepatotoxic of (Macaca of 157:63-69. Bose the K, dioxin WS, Bates Division Jordan do n o t of SW, MN. 1977. Public Ridolpho damage mouse TCDF PF , and Persp; PCB New and human Zealand S m i t h J. marrow isomers 60:77-88. 2,4,5-T Health, bone 6:41-47. Dept 1985. chromosomes. 97 MMWR (Morbidity and Reports. Centers for and Mortality Weekly Serum dioxin Disease Report). 1987. in V i e t n a m - E r a V e t e r a n s Control Morbidity Epidemiologic - Preliminary and Mortality Weekly Notes Report. Report; 36(28):470-474. Mocarelli 1986. P, Marcocchi Clinical children. JAMA; M o h a m a d AH, A, Zerezghi M, JA, Health Moore Gupta BN, exposure Perspect; RW, plasma 81-85. K, JA. 1986. CL, and Yos JG. in T h e o b a l d HM, in male Haworth Orange using (Cited tetrachlorodibenzo-p-dioxin. of Agent of P, Young exposure to DS , Mantel dioxin Determination capillary Anal 1973. gas N. in Chem; of chromatography 58(2): Postnatal S, P e t e r s o n RE. treated with Toxicol Appl Zeiger compounds. of (TCDD). Environ 1985a) R o b i n s o n JA, rats S p e c k W, related EPA 469-471. effects 2,3,7,8-tetrachlorodibenzo-p-dioxin deficiency Mortelmans Gerthoux detection. Z i n k l JG, to 5: Potter Androgenic Caruso dibenzo-p-dioxins with microwave-induced maternal P, 256:2687-2695. polychlorinated Moore Brambilla laboratory manifestations Pharmacol; E. 1984. Toxicol 1985. 2,3,7,879:99-111. Mutagenicity Appl testing Pharmacol; 75:137- 146. Moses M, workers Prioleau with P.G. 1985. and without Cutaneous chloracne tetrachlorodibenzo-p-dioxin. Mottura A, Zei chromosome Seveso accident. M u l c a h y MT. 2(10): Nuzzo FJ, on Mutat 1980. 573-574. ♦Murray 1979. G, analyses FA, J Am Acad et al. findings exposure Dermatol; 1981. to in 1 2 (3>:497-506. Evaluation of results TCDD exposed Res; 238-239. (Cited in "Agent Orange." 85(4): aberrations 1985a) Nitschke KD, Humiston reproduction tetrachlorodibenzo-p-dioxin and (TCDD) study in the of CG, subjects EPA Kociba rats diet. chemical 2,3,7,8- of in E P A Three-generation past lymphocytes Chromsome (Cited Smith F, histologic with Toxicol the 1985a) RJ, given of after Med J Aust; Schwetz BA. 2,3,7,8Appl Pharmacol; 50:241-251. Naikwadi KP, Karasek FW . 1986. Gas chromatographic 2.3.7.8- tetrachlorodibenzo-p-dioxin tetrachlorodibenzo-p-dioxin capillary column. Nagayama J, mediated induction J isomers Chromatogr; Klyohara C, of lymphoblastoid cells 2 . 3 . 7 . 8 - TCDD. Arch by using a polymeric Kuratsune M. 1985. hydrocarbon hydroxylase polychlorinated Toxicol; 56: of liquid and crystal 3 6 9 ( 1 ) :203-207. M a s u d a Y, aryl separation from polychlorinated biphenyls Genetically activity dibenzofuran in hu m a n isomers and 230-235. 5019 'T ) - 6 1 98 N a u H, Bass R, Neubert D. 1986. Transfer tetrachlorodibenzop-dioxin (TCDD) toxicity Toxicol; Nelson the in the mouse. C J , H o l s o n JF , relationship occurrence ♦Neubert Arch G r e e n HG , between in Arkansas. D, Dillman I. 1972. Arch Neubert D, Zens embryotoxic Perspect; Pharmacol; dioxins. Overview N i e m a n n RA. of TCDD Lamparski 1986. of and (National J Department Centers for soil scale data. Assoc Institute of Health Disease and (Cited in E PA in m i c e study cleft of palate 1985a) treated with HJ. 1973. species. survey of the RA , Kocher CW , et environmental survey for chlorinated Hummel Chemosphere; 15: 69(6): of Occupational Safety and Human Current 2,3,7,8- and Public Institute capillary 976-980. Health). Intelligence Services National of electron-capture Chem; al. 1453-1460. determination in fish b y Off Anal Control, A Environ Health 1985a) '-I, 3 , 7 ,8-Tetrachlorodibenzo-p-dioxin. U.S. Retrospective 2,4,5,7-T effects Merker Surrogate-assisted gas-chromatography. 1979. of 2,3,7,8-tetrachlorodibenzo-p- F r a w l e y NN, large tetrachlorodibenzo-para-dioxin NIOSH and in m a m m a l i a n in E PA LL , a and postnatal 2 7 2 ( 3 ) :243-264. (Cited Perspectives and milk, 19:377-384. R o t h e n w a l l n e r A, 5:67-79. N e s t r i c k TJ, 1986. P, effects DW . use Embryotoxic acid 2,3,7,6- 59:36-40. Gaylor Teratology; of placenta agricultural 2,4,5-trichlorophenoxyacetic dioxin. via for 1984. Bulletin Health 40 Service Occupational Safety and Health. ♦Nisbet ICT, studies of P a x t o n MB. the 1982. reproductive Statistical toxicity aspects of TCDD and of three-generation 2,4,5-T. Am S t a t Vol; 3 6 ( 3 ) :290-298. N o l a n RJ, of Smith FA, Hefner JG. 1979. Elimination 2,3,7,8-tetrachlorodibenzo-p-dioxin following a (Cited EPA in Nottrodt against single IA, dose. Ballschmiter emissions (National of of K. Publ. Program, Program. No. NCI, Appl Causes tissue Pharmacol; (NIH) Program). distribution guinea 48(1): pigs A162. Research Bethesda, Triangle and reduction strategies incineration plants- Chemosphere; 1982a. for possible 82-1765. for from waste recent measurements. Toxicology NIH; 1986. PCDD/PCDF tetrachlorodibenzo-p-dioxin DHHS Toxicol and in female 1985a) interpretations *NTP oral (TCDD) Bioassay 15: of 2,3,7,8- carcinogenicity MD: Park, 1225-1237. (gavage Carcinogenesis NC: National study). Testing Toxicology 99 NTP (National Toxicology Program). tetrachlorodibenzo-p-dioxin DHHS Publ. Program, No. NCI, (NIH) NIH; for 82-1757. 1982b. possible Bethesda, Research Triangle Bioassay of 2,3,7,8- carcinogenicity MD: Park, (dermal Carcinogenesis NC : National study). Testing Toxicology Program. N y g r e n M, Rappe C, Identification of Lindstrom G, dibenzofurans in env i r o n m e n t a l G, eds. Keith, LH, Chelsea, MI: Lewis Oehme Mano S, the M, and O'Keefe P, of Meyer C, Chemosphere; Oliver RM . in 1975. 15: Toxic laboratory workers. O l s o n JR. 1986. pp. samples. D, for In: al. Rappe 1986. and C, in 1986. Choudhary Perspective. Quantitative of polychlorinated in o u t d o o r air. Hilker et dioxins 15-24. P. amounts cartridge P A, and Dibenzofurans Kirschmer S m i t h R, adsorbent water. Inc; femtogram dibenzofurans Reverse-phase and human Publishers, Bergqvist polychlorinated Chlorinated Dioxins M i k a l s e n A, determination dioxins H a n s s o n M, 2,3,7,8-substituted Chemosphere; Aidons K, trapping method 15(5): Wilson dioxins for d i h e n z o p- L. 607-617. 1986. in drinking 1127-1134. effects Br J Metabolism of 2,3,7,8-tetrachlorodibenzo-l,4-dioxin I n d Med; 32(1): and disposition in guinea pigs. - tetrachlorodibenzo-p-dioxin 49-53. of (Cited in EPA 1984). 2,3,7,8- Toxicol Appl Pharmacol; 85:263-273. O l s o n JR, Wroblewski V J . 1985. tetrachlorodibenzop-dioxin pigs a n d rats. O l s o n JR, Chemosphere; Gasieqicz excretion, in the Golden in EPA 1985a) O l s o n JR, TA, of TA, R A Neal. Toxicol Geiger LE, and Pocchiari Aspects, O t t MG, Holder engaged in Med; BB, 47-50. O t t MG, O l s o n RA, workers with Med; Ozvacic PCDFs. V. eds. Press; O l s o n R0 . 1980. of from guinea distribution, Appl Pharmacol; RA. 1983. Exposure 81-100. A to ( C i t e d ---- metabolism of systems. Dioxins: (Cited mortality The in in E P A C o o k RR . 1987. exposure (TCDD) 56:78-85. EPA analysis In: Coulston Human Health 1985a) of 2,4,5-trichlorophenoxyacetic (Cited potential Tissue in mammalian Accidental Academic the manufacture 22(1): Occup F, N e w York: 2,3,7,8- 1980. Neal 2,3,7,8-tetrachlorodibenzo-p-dioxin R, of isolated hepatocytes 2,3,7,8-tetrachlorodibenzo-p-dioxin Syrian hamster. Gasiewicz in 14:979-982. Neal, and metabolism Metabolism (TCDD) employees acid. J Occup 1985a) Cohort mortality to the h i g h e r study of chemical chlorinated, d i o x i n s . J 29(5):422-429. 1986. A Chemosphere; review of 15: stack sampling methodology for PCDDs/ 1173-1178. 5021 100 P a l a n s k y J, 1986. Kapila Studies mobility of Patterson S, Manahan on vapor 2,3,7,8-TCDD DG, H o f f m a n RE , persons Patterson LL . in h u m a n 16: DG, 1987a. adipose tissue DG, resolution analysis JAMA; 256: Jr, gas al. 15: TE. on 1389-1396. 1986. adipose Clevenger dispersing medium 2,3,7,8- tissue of exposed and 2683-2686. Boozer EL, Lapeza on whole-weight and lipid basis. H a m p t o n L, Lapeza DG, CR, Jr, e t al . CR , Needham (TCDD) Chemosphere; serum on a whole-weight H o l l e r JS, Anal Lapeza Chem; C R, 59:2000-2005. Jr., adipose tissue 58(4): 705-713. assumptions Toxicol for et Nemcova and prognosis tetrachlorodibenzo-p-dioxin (Cited in Philippe EPA M, FJ . M, of of K r a s n o b a g e V, Z e y e r J, Symp; Arch conditions. Rose RQ, FEMS Gehring Perspect; 5: Pitot PJ . 241-244. (Cited G o l d s w o r t h y T, diethylnitrosamine. Cancer Poiger H, C. dermal and Schlatter intestinal (Cited 1973. Schlatter metabolism *Poiger H, of soil. Lukas Environ Health; C. 15:9-12. C. R. 1981. Reg E. 36(1): Fate Res; of cultures 1981. 5-11. 2,3,7,8- and soil under­ 2210-2233. Excretion in the and tissue rat. Environ distribution Health 1985a) 1980. Promotion by of hepatocarcinogenesis 1980. 40:3616-3620. Influence of of TCDD. (Cited solvents 2,3,7,8- from in and EPA 1985a) adsorbents Food Cosmet Toxicol; on 18(5): 1985a) 1985. of TCDD Schlatter in EPA absorption in E PA hepatic Chemosphere; of human Chem; examination J i r a s e k L, in microbial 12: P o l a n d H. tetrachlorodibenzo-p-dioxin H, critical Anal intoxication by Huetter 2,3,7,8-tetrachlorodibenzo-p-dioxin Poiger A P i c k o v a J, in men. laboratory 477-481. analysis 1985a) (TCDD) HC, High-resolution dioxin contaminated chronic tetrachlorodibenzo-p-dioxin P i p e r WN, 1987c. spectrometric 1986. assessments 2,3,7,8- 6:684-307. P a z d e r o v a - V e j l u p k o v a J, development Murray in r i s k Pharmacol; al. for ... 2,3,7,8-tetrachlorodibenzo-p-dioxin. D J , S h u HP, used High- spectrometric and lipid basis chromatographic/high-resolution mass Paustenbach, 1987b. chromatographic/high-resolution mass of human Patterson of of 2,3,7,8-tetrachlorodibenzo-p-dioxin tetrachlorodibenzo-p-dioxin. The et in E e l s o n WT, of M a l h o t r a RK, role 935-936. Patterson gas LL, levels H o l l e r JS, Determination AF, and Chemosphere; Needham in Missouri. Jr, Vonders transport in soil. Tetrachlorodibenzo-p-dioxin control SE, phase in 1986. Influence the dog. of phénobarbital Experientia; Pharmacokinetics of and TCDD on the 41:376-378. 2,3,7,8-TCDD in man. 101 P o i g e r H, Weber kinetics of TCDD metabolite(s) Pocchiari the Schlatter in in F, dogs guinea eds. pigs. N e w York: Glover E. 1973. gamma-aminolevulinic hydroxylase. Pharmacol; II . A 9:736. P o l a n d A, binding Glover of ribosomal CH. 1982. a n d rats. Special In: Hutzinger of 0, Frei and Merian (Cited synthetase of in EPA 1979. the An and structure aryl and TCDDE, Impact in EPA Chlorinated dibenzo-p-dioxins: Potent acid study DNA. RW, of Compounds. 317-325. on 1985a) inducers hydrocarbon activity relationship. Mol 1985a) estimate of the m a x i m u m 2, 3 , 7 ,8-tetrachlorodibenzo-p-dioxin RNA of metabolism toxicity and Related P e rgamon Press; (Cited E. aspects Assessment Chlorinated Dioxins Environment. P o l a n d A, of H, C a n c e r Res; 39(9): to in v i v o rat 3341-3344. covalent liver protein, (Cited in E PA 1985a) P o l a n d A, Glover segregation P o l a n d A, of E. 1980. K n u t s o n JC. related halogenated of toxicity. Pola n < t A P , workers Rev Smith D, chloracne, 2,4-D Metter and 22: P o l a n d A, Greenlee action the of Sci; P o l a n d A, mice. Potter CL, D, K e n d e AS. Glover 1979. (Cited A 17(1):86-94. E. 1982. (Cited health and mechanism in EPA survey attention 1985a) of to parameters. Arch on and the mechanisms of related compounds. Ann Tumour promotion by (Cited 1986. in EPA TCDD in s k i n of 1985a) Relationship of alterations in 2,3,7,8-tetrachlorodibenzo-p- 6:89-97. Ertl DC, R e i s n e r RM. study of cutaneous established the 1985a) 271-273. treated with of 1984). Studies in E P A P e t e r s o n RE. S a k a m o t o M, chloracne: of A special dibenzo-p-dioxins 300(5889): Toxicol; application 1971. in EPA dioxin. SM, P. and psychologic M e n a h a n LA, for Possick 22:517-554. (Cited in rats ♦Puhvel G, Toxicol; Examination tarda, WF, Nature; Pharmacol; cutanea energy metabolism models Mol 316-327. 320:214-230. Fund Appl locus. 2,4,5-T plant with chlorinated Palen HRS/J the A h 2,3,7,8-Tetrachlorodibenzo-p-dioxin Pharmacol porphyria Environ Health; NY Acad 1982. aromatic hydrocarbons: Ann in a 2,3,7,8-tetrachlorodibenzo-p-dioxin toxicity with 1982. Hairless changes chloracnegens. Toxicol mice induced by Appl as topical Pharmacol; 64:492-503. Putoni in R, Merlo Seveso. Rappe C. 1984. PCDDs and 135 Environ Rappe Sci C, exposure Perspect; F, Lancet; Analysis Santi of polychlorinated can be Technol; 18(3): S. identified by L. 1986. Soft tissue sarcomas dioxins and furans. isomer-specific All 75 techniques. 78A-90A. Marklund, polychlorinated 60: M e a z z a L, 2:525. PCDFs N y g r e n M, to F i n i A, et al. 1985. dibenzofurans and Assessment dioxins. of human Environ-Health 303-304. 5023 102 Rappe C, Marklund emissions Reggiani Toxicol S, Kjeller from various G. T y s k l i n d M. Acute human exposure 6(1): 27-43. S i s k o A, PCDDs to TCDD (Cited in and Seveso, in E PA Italy. S. 2,4,5-trichlorophenoxyacetic herbicide in Finland. in EPA ♦Roberts EA, Scand J Work Manchester DK. S h e a r NH, toxicity: Rogers of 2,4- AM, O k e y AB, From rodent A n d e r s o n ME, to h u m a n B a c k KC. 1985. tissues. 1982. The Ah Mutagenicity of in E P A lymphoma cells. Kornel P C B s , dioxins, JF, A, and Research Treatment Robens B. R o z m a n K. n.d. Land (Cited Mobile Disposal, Triple Thermal 1987. Wastes, residue. Chemosphere; KPEG quadrapole Michigan properties of 15: 1325-1332. Separation of was t i n g 1985a) destruction Action. OH. mass State in L 5 1 7 8 y Presented Remedial Cincinnati, May and 14:661-674. at unit the for 13th Incineration 6-8, 1987. spectrometry and RM temp University. dioxins, furans and related syndrome and 2.3.7.8- tetrachlorodibenzo-p-dioxin. Toxicol Lett; R o z m a n K, 2,3,7,8-tetrachlorodibenzo-p-dioxin in 1984. 105:445-449. in c o n t a m i n a t e d wastes. Symposium: 1986. Res; P e t e r s o n RL. of Hazardous Z a b i k M. compounds. Mutat furans phosphoresence-dioxin Rondorf 37-42. 2,3,7,8- mouse and 8: receptor Chemosphere; acid Annual acid Environ Health; tetrachlorodibenzo-p-dioxin and perfluoro-n-decanoic CJ , J 1985a) dioxin Rogers in 1985a) acid applicators Mortality PCDFs 1213-1218. Hemberg and 1982. 15: dichlorophenoxyacetic (Cited V, 1986. Chemosphere; Environ Health; Riihimaki 1980. LO , incinerators. G r e i m H. cold-adapted R o z m a n K, hormones Health; Ryan JJ. Canada. 1986. rats. R o z m a n T, modulate 16: Toxicity Arch Scheufler the of Toxicol; E, toxicity 1987. Food Research Division, communication NY . September 2, 1987. R y a n JJ, PY, P i l o n JC, et al. 1984. 2.3.7.8- tetrachlorodibenzo-p-dioxin Sci Health telephone Corporation, Environ T, H 2,3,7,8-TCDD. Greim. J 1985. Toxicol Thyroid Environ 481-491. Personal Lau caused by 22:279-285. 59:211-215. Pazdemik of lethality Technol; 18(9): to Protection Branch, D K Basu, Incidence in Lake and Ontario Syracuse levels Ottawa, Research of commercial fish. 719-721. / ♦Key s t u d y R y a n JJ, Lizotte R, Sakuma dibenzo-para-dioxins, chlorinated Canadian chicken and pork samples. T, Mori B. dibenzofurans J Agric Food 1985ba. Chlorinated and pentachlorophenol Chem; 33(6): in 1021-1026. 103 R y a n JJ, Schecter distribution population. R y a n JJ, Lizzote dioxins and Chemosphere; 14: Lizotta chlorinated 14: of A, R, R, furans Miller in h u m a n s G A, 2.3.7.8bean 1985bc. from Chlorinated in Canadian h u m a n Vigano and maize SANSS P, Fortunati G, Cocucci tetrachlorodibenzo-p-dioxin plants. (Structure printout, April S a w a h a t a T, the Tissue general dibenzo-p-dioxins adipose Experentia; 42(5): and Nomenclature Search 3, isolated tissue. and Chemosphere; O l s o n JR, EPA S c h e c t e r A, chemical Tieman analysis Neal T, and RA. 1982. Ryan dibenzofurans populations. Dioxins Inc.; Schroy J M, Schaffner exposure Persp; (TCDD) formed Res F. 1985. of metabolites on of incubation with Commun; 105(1): 341-346. fat b i o p s i e s to p o l y c h l o r i n a t e d G. in h u m a n C, Patient for for ultrastructural 1986. adipose Choudhary in Chlorinated tissues G, Keith d i o x i n s , furans and Perspective. dioxin and from LH, exposed eds. Chilson, 1957. der Cheng Klinishce chloracne. in E P A BA , FD , Chemosphere; SC. 14: and control Chlorinated MI: Lewis Publishers und 1985. Physical/chemical experimentelle Arch Klin Exp Norris JM, S p a r s c h u GL, et al . S e e f e l d MS, P e t e r s o n RE . Digestible in rats S e e f e l d MS, of 1984. treated with Pharmacol; Characterization 74: zur 589-596. (Ger.) Keesey treated with 76: RE, Toxicology Persp; energy and 5: of 87-99. efficiency of feed 2,3,7,8-tetrachlorodibenzo-p-dioxin. 214-222. Corbett the w a s t i n g tetrachlorodibenzo-p-dioxin. Pharmacol;. 206: 1973. utilization S e e f e l d MS, of 1984). Environ Health study untersuchungen Dermatol; dibenzo-p-dioxins. Appl properties 877-880. chlorinated Toxicol On-line 60:241-254. Gitlitz Rappe Hileman KH. *Schwetz 1987. 51-56. atiologie (Cited JJ, levels I n: 2 . 3 . 7 . 8 - TCDD. Schulz System). Biochem Biophys and Dibenzofurans pp. of solution by 586-588. Identification liver biopsies after Environ Health S c h e c t e r A, Accumulation and nutrient 1985a) characterization PCBs. 1986. soil 1987. rat hepatocytes. (Cited in SM . from 2.3.7.8- tetrachlorodibenzo-p-dioxin rats 1985bb. 697-706. Sacchi *Key L. 929-932. L a u BPY. dibenzofurans S u n WF, SW , Keesey syndrome RE, in Toxicol Appl P e t e r s o n RE. 1984b. P e t e r s o n RE. rats Pharmacol; Body 1984a. treated with 73 : weight 2,3,7,8-tetrachlorodibenzo-p-dioxin. 2,3,7,8- 311-322. regulation Toxicol in Appl 526-536. X L 5 1 ^ 5025 104 S e i l e r JP. 1973. Experiencia; S h i v e r i c k KT , (TCDD) S h u HP, use Paustenbach assessment S l a g a TJ, DJ, 7: NJ; 2, 3 , 7 ,8-tetrachlorodibenzo-p-dioxin microsomal steroid metabolism Biochem Pharmacol; Murray pesticides. F J . 1987. and A 32: and serum 991-995. critical evaluation tumor promotion 2,3,7,8-tetrachlorodibenzo-p-dioxin. of the data in a Reg Toxicol cancer S. 1985. Senear mouse M i l l m e n HA, skin t u m o r i g e n e s i s . In : Weisburger EK, Eds. Noyes Handbook Pub. Park 230-250. Pearce (F R G ) . (Cited S m i t h AH, NE. 1985. Dioxins Fisher DO, 37:197-200. (Cited Fisher sarcoma phenoxyacetic et in DO, at the 5th 1985a) Giles HJ, Pearce study: exposure. 16-19. Symposium Bayreuth 1986) 2,4,5-T EPA International C o m p o u n d s . Sept. C h a p m a n C J . 1982. Zealand case-control acid al. Dip NP, among New S m i t h AH, Presented and Related in Hiremath miscarriages Congenital sprayers. N. 1983. Interview Chemosphere; Arch The New findings 12(4/5): defects Environ and Health; Zealand soft concerning 565-571. (Cited in 1985a) Smith FA, Schwetz BA, Nitschke tetrachlorodibenzo-p-dioxin 517-523. (Cited S m i t h RM, 0'keefe picogram per in E P A high-resolution Anal Chem; Sparschu GL, Appl of 2,3,7,8- Pharmacol; 38(3): DR , Aldous KM. of and 1986. Determination tetrachlorinated dibenzo-para-dioxins in of and indoor air by 2414-2420. FL, 405-412. FL , Lisowe Food Cosmet Toxicol; SRI (Stanford Research United Toxicol concentrations Jr, R o w VK, (Cited RW, 2 ,4,5-trichlorophenoxyacetic Producers. Teratogenicity mice. Jr. 1971a. Study 2,3,7,8-tetrachlorodibenzo-p-dioxin 9: Dun 1976. chromatography high-resolution mass-spectrometry. J r . , Dunn Toxicol; S p a r s c h u GL , Hilker dibenzofurans of KD. in CF-1 1985a) meter gas 58(12): teratogenicity Cosmet PW, cubic pentachlorinated of of various 1985a) 57-88. Chlorinated EPA 1983. in EPA carcinogenesis, of Nesnow S m i t h AH, -tissue the muta g e n i c i t y (Cited rats. Carcinogen Testing. Rig, on TF . of pregnant Pharmacol; on on hepatic of mutagenesis, risk of survey 622-623. Muther effects estradiol A 29: 9(40; R o w e VK. acid 527-530. Institute). States In E P A on 1987. SRI the the rat. Food 1985a) 1971b. fetal (Cited of America, of in Effects in E P A 1987 of high development in the levels rat. 1985a) Directory International, of Chemical Menlo Park, CA. 105 Stalling DL, Residues of p o l y c h l o r i n a t e d Laurentar Smith Great LM, P e t t y JD, Lakes fish. Chlorinated Dioxins Eds. Publishing Plenum Stalling pattern GC/MS DL , S t a n l e y JS, PCDDs and Stehr PA, possible dioxin Scand^J Sci; Tenchini A ML, Tucker N o r s t r o m RJ, of Sack tissue PCDD TM, AM, following et al. R o t h W. 1986. A Working June. 1986. Use residue JC, et EPA TY82 of data from al. 1986. NHATS ash. Chemosphere; "the hallmark 11: 165-171. Environmental epidemiologic E n viron Health; chloracne: of study 41(1): different 15: dioxin Update C, Pacchetti cytogenetic G, of 16-22. treatment 1135-1140. intoxication." and R. M o t t u r a A, study 1977. on overview. occurred Joint NIEHS/IARC (Cited in EPA Ann NY DeCarli L. E E, Moore infection as JA. a 1980. Internal TO , DC. T a y l o r ML, polychlorinated Karasek, ionization, 15: G a r r e t t JH, 1986. P. samples electron on exposed 1 3, to November, Lyon, ------- 1975. Office exposure 12(6): of Toxic et al . 1985. Pers; of 59: Sources and to 1319-1324. Substances, by high fate of compounds in 145-158. quantitation resolution and mass and related and polychlorinated capture Increased of 1985a) Comparison dibenzodioxins environmental Chemosphere; to in E P A Environ Health FW . in 73-85. Report, I m m u n Dec; dibenzodioxins, dibenzofurans environments. polychlorinated memo (Cited BASF Group sequela Cohn, H F. 5: abortions of persons the Working Thomas Washington, study in S, induced 1985a) McConnell of bacterial Agosti of Environ Mutagen; Mortality an accident which F a i t h RE, cases Infect flame the pilot 2,3,7,8-tetrachlorodibenzo-p-dioxin. complex S i m o n M. Investigation of Papers, susceptibility HY, G r a y AP , 2,3,7,8-tetrachlorodibenzo-p- Arch 1986. in fly Frentzel-Beyme T h i g p e n JE, Tong in of Y o u n g AL, and PCDF Remmers from associated with and non-exposed women. dioxin human E, Risks 15:1605-1612. in Missouri. Crimaudo Thiess Tieman 1983. 320:295-307. TCDD-exposed EPA, al. 221-240. adipose Chloracne, comparative France, LM, et dibenzofurans 15:1435-1443. Environ Health; 1979. p. and Environmental Compounds. NY; O n s t o t J, PCDD/PCDF 1985. Work Johnson JL and evaluation F a l k H, effects Z w i c k G, for - - T a y l o r J S. 1953. KE, in human contaminations L, Human Smith the Chemosphere; S u s k i n d R R. 1983. PH, in Boggess S t e i n G, techniques Corp., Chemosphere; health Stieglitz Acad Peterman PCDFs repository. In: and Related recognition analysis. H o g a n JW, dibenzo-p-dioxins of dibenzofurans gas in chromotography spectrometric with detection. 1141-1146. 5027 106 T o n g HY , Shore DL , Karasek dibenzodioxins and polychlorinated mixture by two-step quantitative T o t h K, dioxin liquid analysis. Somfai-Relle of herbicide Isolation Chem; Sugar J, Olah mice. E, Nature; 56: Bence S, J. in Giri ♦ T u r n e r JN, fluid 67: U m b r e i t TH, soil from Umbreit from a ymbreit or (Cited testing and in EPA of p u r e 1985a) of herbicide dioxin on mutation Carcinogenesis; and 5:1725-1728. Liver morphology products of in guinea pigs a polychlorinated biphenyl 2,3,7,8-tetrachlorodibenzo-p-dioxin. D, Hesse TH, 1983. G a l l o MA. industrial 2,4,5-T 548-549. dioxin Toxicol Appl 417-429. Patel an TH, DN. either pyrolysis transformer Pharmacol; Carcinogenicity containing 1986) Collins administered organic for Effect exchanges. chromatid complex 1984. induction sister a containing (TCPE) (Cited 1979. S u g a r J. 2,4,5-trichlorophenoxyethanol of for fractionation 2442-2447. 278(5704): Somfair-Relle of polychlorinated dibenzofurans chromatographic Anal S, 1984. 2,4,5-trichlorophenoxyethanol in Swiss T o t h K, F W. EJ , site. G a l l o MA. manufacturing Hesse contaminated soil Chemosphere; 15: 1985. E J , Gallo from 1986a. site. MA. Times Acute toxicity Chemosphere; 14: Bioavailability Science; 1986b. Beach, of TCDD 232: of dioxin in soil 497-499. Comparative Missouri, contaminated 945-947. and toxicity Newark, of TCDD New Jersey. 2121-2124. Van Miller JP , neoplasms in rats Lalich JJ, A l l e n Jr. exposed to low tetrachlorodibenzorho-dioxin. 1977a. levels Increased of incidence of 2,3,7,8- Chemosphere; 6(10): 625-632. (Cited in EPA 1985a) Van Miller JP, Lalich JJ, neoplasms in rats p-dioxin. Chemosphere; Velzy 1986. CO. A l l e n JR. exposed ASME to low 6(9): Standard JG, Moore, JA, Zinkl Environ Health W a l d e n R, Persp; Schiller CM. 5: Toxicol Appl W e b e r H, and Bimbaum 1985. the to the and Effect Comparative 1985. 77: in in EPA analysis 1985a) methods for of 2,3,7,8- system of laboratory animals. embryo toxicity four of 2,3,7,8- (sub)strains o£ adult 2,3,7,8-tetrachlorodibenzo-p-dioxin and male 490-495. 2,3,7,8-tetrachlorodibenzofuran Distribution of 2,3,7,8-tetrachlorodibenzo- (Cited immune (TCDD) Pharmacol; LS. of incidence 1179-1185. 1973. on Increased 149-162. tetrachlorodibenzo-p-dioxin rats. sampling 15: JG. tetrachlorodibenzo-p-dioxin levels 537-544. dioxins/furans. Chemosphere; ★Vos 1977b. (TCDF) excretion. in p r e gnant Arch Toxicol; C57BL/6N 57: (TCDD) mice: 159-162. 107 Weber H, Poiger metabolites Weber of H, Harris TCDD, Lett; H, Schlatter in m a l e TCDF, guinea MW, C. 1982. pigs. H a s e m a n JK, and TCDD-TCDF NCA, Birnbaum veterans seeking medical W e e r e n RD, its Wolfe Asshauer esters. WH, 707-716. J Lathrop (Cited Wroblewski J. P a n JC, assistance. 1985. VJ, Assoc GD , Albanese in H i r e m a t h O l s o n JR. Pharmacol; Wurrey'CJ, 81: Bourne Young S, 1985. 1985. TCDD- Teratogenic mice. potency Toxicol 1984. Young AL, studies Young AL, Kang JA, HK, Zeiger E. results 22(1): Sci RD. PM. rat 1984. and of Chemosphere; of acid 14: 2,3,7,8- g u i n e a pig. Application 11-14. and Toxicol the Zimmering S, M a s o n JM, Valencia testing D ro so p h ila . National EPA f r o m Dr. NIEHS. in of human of 33(6): federal 530A-540A. in EPA R, Toxicology spectrometry exposure to the 702-709. epidemiologic 14 : 779-790. dioxins as h e r b i c i d e __ (Cited in EPA experience trichlorophenol in gas 482-83. Chlorinated 17: of infrared Chemosphere; mortality in a for results TCDD. 1983. The (Cited 58(2): Contam Toxicol; to (Cited performed transform and measurement BM. 1980. test the analysis 917-921. metabolism in the Chem; NTP, for 68(5): 1986. Program. tested trace Chem; Technol; Memorandum mutagenesis of Hepatic Status 1983. of and results 1986) exposed Shepard Environ S u s k i n d RR. Med; 2,3,7,8of Vietnam 1787-1794. Moynahan Anal tetrachlorodibenzodioxin Occup of tissue 15: al. Environ 1985. populations contaminants. Levels in 2,4,5-trich l o r o p h e n o x y a c e t i c (TCDD) Determination K a n g HK. of 1986. in adipose Chemosphere; isolation/fourier determinations. AL. al. RA, et Kleopfer d i benzo-p-dioxins. Bull to of 117-122. 231-240. chromatography/matrix dioxin et Problems Off Anal tetrachlorodibenzo-p-dioxin Zack toxicity 14: in C 5 7 B L / 6 N (2,3,7,8-TCDD) 2,3,7,8-tetrachlorodibenzo-p-dioxin to L S. combinations S c h e c t e r AJ, tetrachlorodibenzo-p-dioxin Appl oral Lett; 26:159-167. Weerasinghe and Acute Toxicol of workers process 1985a~) exposed accident. J 1985a) Zeiger to Dr. Environmental E.E. McConnell Mutageneis on th e Development 1985a) W o o d r u f f RC. II. Results Program. 1985. of 20 Chemical coded compounds Environ Mutagen; 7: 87-100. t 502 9 ^ *? |3 0 ^ 0 3 0 11. Acute Exposure--Exposure less, as specified Bioconcentration chemical time at the time Carcinogen--A chemical Ceiling value (CL)--A exceeded, Chronix even in conception (either the time of at the and two Effect severity in advisory technical a of a 14 or the concentration of or during time in a the a discrete surrounding period. substance that should n o t be chemical for result of from life to the a toxic chemical; growth, and in útero a guidance Dangerous 30 m i n w i t h o u t health effects. in on the chemical or prior to postnatally effects may be organism. effect the on the conceptus distinguishing as a feature during which include the malformations ------ and death. exposure effects, an which increase such as exposed of acceptable enforceable federal, to Life any a development, development of on health assist as in produces a frequency or irreversible population when control. estimate concentration within level or. m o r t a l i t y , legally to of significant adverse to the stage appropriate more, effects developmental of u s e d here, based adverse span as its or Adverse is (FEL)--That days exposure during prenatal the 365 Profiles. occurrence or b i o l o g i c a l l y is n o t Immediately of o f .inducing c ancer. terms, substance environmental days concentration same The impairment chemical to exposure EPA Health Advisory--An a the concentration of unmistakable functional time terms Level compared with of specific Fetotoxicity--Any altered statistically a duration Profiles. quotient the s e x u a l .m a t u r a t i o n . occurred. variations, Frank capable parent), of prenatal between insult during that may any point Embryotoxicity result a divided by or Toxicity--The organism to at Toxicological developing detected for instantaneously. the Developmental chemical (BCF)--The Exposure--Exposure specified a Toxicological organisms exposure same to the Factor in aquatic period of water in GLOSSARY of or a effects federal state, Health drinking water information. standard, and local (IDLH)--The contaminant escape-impairing for but serves as officials. maximum from which symptoms levels A health or one could escape irreversible 5031 110 Intermediate days, as Exposure--Exposure specified Immunologic system in the result a chemical for a duration Profiles. occurrence of adverse to environmental Toxicity--The that m a y to Toxicological from exposure effects on of 15-364 the agents immune such as chemicals. In v i t r o - - I s o l a t e d as in a test from In vivo--Occurring Key Study--An the n a ture those living organism and artificially maintained, within animal of the the living or h u m a n adverse organism. toxicological effects produced study and that best the doses illustrates associated with effects. Lethal in the tube. Concentration(LO) air which has been (LCLo)--The reported lowest to h a v e concentration caused death of a in humans chemical or animals. Lethal Concentration(50) chemical in air expected to cause (LCso)--A to w h i c h death exposure in 50% calculated for of a a concentration specific defined length of experimental of a time is animal population. . Lethal Dose(LO) route other humans or Lethal Dose(50) to cause (LDLO)--The than lowest inhalation that is dose of (LD50)--The in 50% of a defined Lowest-Observed-Adverse-Effect in a biologically effects study or between or group significant group significant the of Level increases in and development, Minimal Risk that likely Level--An (noncancerous) Mutagen--A the genetic defects, chemical to chemical have introduced by caused or substance material a death a in lowest produces its been or dose severity dose of statistically severity of of statistically appropriate lowest calculated population. of or adverse control. chemical in a or b i o l o g i c a l l y effects between the control. changes that may adversely affect function. of daily human an exposure appreciable risk duration exposure. causes in a body or or structural specified miscarriages, and produces estimate that animal in f r e q u e n c y appropriate to b e ' w i t h o u t over which ( L O E L ) - -The frequency which has (LOAEL)--The studies which its Malformations--Permanent survival, a Level of increases studies exposed population is a experimental exposed population Lowest-Observed-Effect study of expected animals. death chemical dose mutations. cell. cancer. of A Mutations of mutation can to a chemical deleterious lead effects ' is a change to b i r t h in Ill Neurotoxicity--The occurrence following to exposure a of adverse effects No-Observed-Adverse-Effect Level there or biolo g i c a l l y are no frequency dose, statistically or population but severity are or no appropriate they are not severity of Permissible Exposure air (NOEL)--That seen between Limit averaged an upper-bound estimate curve determined by calculate cancer food, an risk per unit and pg/m^ Reference order Dose for effects hazard of during a of of RfDs additional an are not of is likely The the or (RQ)--The selected for under are Toxicity--The system toxicity may be The in sexual modifications this that directed system. in chemical at w h i c h there increases exposure slope procedure. is of The the in this frequency and its level in the q^* dose-response can be incremental for water, used to excess mg/kg/day for other a of the such functions as an Sect. used on to an a the of estimate a chemical. The RfDs cancer. a hazardous substance quantities are: t h a t — i s --(1) amount established 311 the of 1 lb by Clean Water 24-h period. of adverse from of such fertility, that effects exposure reproductive manifestation behavior, data the from application is b a s e d Reportable or under derived which on to deleterious of database occurrence to of types substances, result risk population consistent effects a spanning perhaps the h u m a n operationally by factor, over may of without quantity CERCLA reproductive alterations RfD CERCLA. measured Reproductive endocrine of low-dose to b e to n o n t h r e s h o l d or either at (with uncertainty entire under Quantities exposed produced allowable exposure modifying of reportable regulation the in may be exposed population reflect various Quantity Act. increases shift. studies) considered (2) at which (usually p g / L daily Reportable greater chemical adverse. the the estimate the that judgment applicable 8-h exposure and h u m a n factors and of carcinogenic potency, lifetime. uncertainty professional dose significant air). that (from animal system seen between dose the multis t a g e (RfD)--An of magnitude) potential NOAEL estimate nervous significant (PEL)--An over qj*--The as the Effects to b e or biologically effects control. effects control. considered Level appropriate (NOAEL)--That adverse its statistically workplace of and No-Observed-Effect on chemical. are to a on organs and/or toxicity may pregnancy dependent the chemical. the be noted outcomes, on the The related as or integrity of system. t Short-Term Exposure workers can be.exposed excursions between Limit are allowed exposure (STEL)--The for up per day, periods. The to maximum 15 m i n and concentration continually. there must daily TLV-TWA may be at not No least be to w h i c h more 60 than min exceeded. 5033 four 112 Target on Organ target extending assumed Toxicity--This organs from over Teratogen--A development Threshold most those a chemical an Limit workers expressed arising lifetime of as Value a TWA, as a normal Factor experimental variation in through causes (TLV)--A exposed Average from that a STEL, (TWA)--An or (UF)--A data. factor UFs are among adverse as used the uncertainty in e x t r a p o l a t i n g data rather to 10. defects extrapolating exposure, effects to those that affect the than NOAEL data. a substance The to w h i c h TLV may be concentration workweek. to from (4) Usually account of animal and of effect. exposure members the LOAEL exposure in operationally intended (3) equal limited adverse cardiovascular) CL. or 40-h in lifetime a allowable 8-h w o r k d a y sensitivity than single of renal, a chemical. concentration without uncertainty less range (e.g., structural the of a to (2) is a broad systems of e x p o s u r e can be averaged Uncertainty covers organism. Time-weighted over term or p h y s i o l o g i c a l the deriving (1) the RfD the the h u m a n population, data data for to the obtained case of humans, in study uncertainty each of these a that in u s i n g factors is set APPENDIXES 5035 2$ 9 ' 5 ' APPENDIX A: A peer p-dioxin Herbert of review panel was ( T C D D ) . The Cornish, Cincinnati New York have at Medical knowledge of were selected specified Section A peer in in in joint Dr . Shane James Olson, a n d Dr. of Medicine. 2,3,7,8-TCDD's physical e n d points, conformity with the These and Que Hee, State experts chemical mechanisms risk tetrachlorodibenzo- following members: of Dr. University University of collectively properties, action, to h u m a n s . conditions Superfund Amendments panel reviewers' their A into cited scientists listing the exclusion, compound. documents of comments the-profile. this of Michigan; and quantification of the 2,3,7,8 the All for peer human and reviewers review and Reauthorization Act of 1986, 110 . incorporated __ f o r for of Center; School key health exposure, assembled consisted University Buffalo, toxicokinetics, animal panel PEER REVIEW A of from ATSDR and determined which the p e e r profile, exists list are and as with databases also included reviewers' a brief part of of the this 115 reviewed will comments explanation of administrative reviewed in EPA has comments and a record. list be the included not the rationale record for of unpu b l i s h e d Ì>-5Ì31 APPENDIX B: DEPARTMENT AGENCY FEDERAL REGISTER ANNOUNCEMENT OF FOR TOXIC HEALTH AND SUBSTANCES ENVIRONMENTAL AGENCIES: Toxic Protection Agency ACTION: The Law by Human Registry Superfund Amendments hazardous amends and certain Disease Services (DHHS): Agency for (ATSDR); and Environmental substances which Priorities requirements a mandate is toxicological priority profiles list which both potential threat Register on April of are (CERCLA or (ATSDR) most List for of Superfund) for (42 U . S . C . the A g e n c y DHHS and Response, for EPA with found at (NPL). Among these statutory the Administrator each substance 100 chemicals. The list to h u m a n h e a l t h . This FR as to the m o s t et Toxic regard on to the prepare included identified list was 12866) 9601 facilities of A T S D R previously determined posed (52 (SARA) Environmental commonly for 1987 Reauthorization Act requirements Agencies 17th, and Comprehensive Registry CERCLA National chemicals the and Liability Act establishing Substances first and TOXICOLOGICAL PROFILES (EPA). 99-499) - Compensation, seq.) OF Notice. SUMMARY: (Public of Health and Disease AGENCY FRL-3269-7) OF AVAILABILITY Department Substances SERVICES DISEASE REGISTRY PROTECTION ( A T S D R - 2; NOTICE HUMAN AND on the the first 100 significant published required by in the SARA Federal section 110. This 25 draft notice announces toxicological the profiles expected for availability review and dates of the first comment. 5039 117 118 AVAILABILITY: The to b e available publicly following by draft the toxicological date Date/Proflie October 17, profiles CAS 1987: 56-55-3 Benzo(a)pyrene 50-32-8 Beryllium 7440-41-7 Chloroform 67-66-3 Chromium 7440-47-3 Chrysene 218-01-9 D i b e n z o ( a ,h ) a n t h r a c e n e Heptachlor/Heptachlor 53-70-3 epoxide 76-44-8 / Nickel 7440-02-0 N-Nitrosodiphenylamine 86-30-6 29, Aldrin/dieldrin 309-00-2 7440-38-2 B e n z o ( b )f l u o r a n t h e n e - Aroclor . 205-99-2 1248, 11096-82-5, 11097-69-1, 12672-29-6 1242, 1232, 1221, 53469-21-9, 11141-16-5, 11104-28-2 Tetrachlorodibenzo-p-dioxin November 5, 12674-11-2 1746-01-6 1987 Benzene 71-43-2 Bis(2-ethylhexyl)phthalate 117-81-7 Cadmium 7440-43-9 1,4-Dichlorobenzene 106-46-7 Methylene 75-09-2 chloride 1987 Cyanide 57-12-5 Lead 7439-92-1 Tetrachloroethylene 127-18-4 Trichloroethylene 79-01-6 Vinyl 75-01-4 chloride 60-57-1 1254, 2,3,7,8- 30, / 1260, 1016 November 1024-57-3 1987: Arsenic PCRs expected # Benzo(a)anthracene October are indicated: 119 A full 90-day profile, starting period each for public from draft comment the period will be actual release profile will be provided date. The indicated on for close the of each the front of comment each profile. Requests for Ms. draft Georgi Director, Agency 28 the case Five above copies address by of profiles for Toxic Substances Monday „-data 4770 through submitted profiles to: Registry : - you wish free delays, the and Disease except in response the review. charge, to sent period. for public Registry NE, One as will be should be comment available Buford Highway, Friday, to of requestors comments end of will be should bear sent 30333 all the draft \103, and Disease > forwarded, the Room ; GA of undue should be External Affairs Substances South the profiles requested will be In of C l i f t o n Rd. Atlanta, Specify profiles Jones Office for Toxic Chamblee 1600 toxicological copy to Ms. All at GA, from the 8am 28 to comments draft the comments inspection at Written control Jones written this and available. notified. legal holidays. docket each profile (ATSDR), Building Chamblee, notice of they become and the A g e n c y South, 4:30pm, and other toxicological number ATSDR-2.^ 5041 120 SUPPLEMENTARY INFORMATION: I. On October 17, 1986, and Reauthorization Act amends the Comprehensive Liability Act of 1980 Section 110 of requirements for the in o r d e r and (3) President 1986 SARA or amends Law Response, section of: to fill 42 of lists gaps which extends Compensation, 9601 et and and seq.). CERCLA by establishing of hazardous substances profiles data Superfund Amendments U.S.C. 104(i) (1) toxicological research program the 99-499), Superfund, preparation (2) signed (Public Environmental (CERCLA of priority, a the of BACKGROUND of those substances, associated with the substances. In compliance with published on April hazardous substances. into groups four subject of the of first spells out must be of set the list at of a than and profile is of Agencies' define is 1987 internal technical peer further time the given by to the development of of on of year of of CERCLA they first enactment are with the the by which the the 100 down to be of to be the total 100 SARA amendments. but no less often examination, information summary and are to b e the must include the of development. process quality and used filled will to the the the h e a l t h to b e a effects our of — The identify improve provided profiles are the Notice the priority ( 52 FR the key ability to been public The We public profiles review of of to are Although deadline, EPA. list and in These 100 availability we and in the dates of undergone and announcing their comment the and the profiles believe that comment scientific have scientific now participation profiles. States and documents subject experts. the prepared 12870). projected profiles. to to b e developed by ATSDR and have review ascertain and on are 1987 to also announces 17, used substance epidemiologic levels. The outside and information along with these October peer in exposure review of an for profiles or encouraging development completed The toxicological review by and data adequate Register notice extensive and guidelines Federal include exposure profiles April availability to public. the draft list 104(i)(3) necessary information be published 25 as when human the current per enactment EPA development timetable year that with This the seventy-five this were first one that guidelines the 25 w a s substances twenty-five the of i.e., the and further b r oken Section needs accordance 17, 25 toxicological human toxicological to group and remaining is m a n d a t e d required testing available least after available intention first ATSDR first priority 100 was profiles. at The CERCLA, requirements, profiles least effects. significant The made at of whether toxicological of completed within information is the list The of years. significant substance on available associated health each of 12866) the these 1987). three e v a luations. This determination of four years Each of of republication interpretation FR chemicals. to b e rate every levels (52 toxicological 17, once 104(i)(2)(A) priority Profiles were completed within Revision 25 content (by^October completed 1987 This second phase developed. priority SARA the 17, section is on will the not be extra important to merit. 5042 121 Although we of the 25 are reasonably substances were confident considered Federal R e g is te r process, this including unpublished data, notice which that during the solicits may aid key studies the p r o f i l e any the for each development significant revision of studies, these draft profiles. I I. The setting presented of LEVELS of a unique a hazardous respect causing a to group term. Since time to (where of a describe to w h i c h format for exposure have type of has been situations of effect is the u ser significant displaying and the is u s e d effects may be public comment effects use by in of the is to over a diverse interpret the appropriate at effects occur may to make this determinations in any particular levels level level may be in which over which to' a l l o w that chronic for considered exposures has specific symptoms gradual, intended l e v e l s , it w a s exposure a insignificant with debilitating are of in which that may be causing different developed encourage context profiles range graphically the immediately to significant human significance level available), and ranges-over which We low these the of EXPOSURE The on respect specific data are levels depends acute, of people who significance as example, significant with longer SIGNIFICANT HUMAN of problems. substance For highly specific set evaluated. OF instance. A significant human profiles to present the observed. and recommendations on this specific '■'issue. III. We of the are soliciting toxicological published on April priority list comments and the As the we health scope and There toxicological to provide scientists that in to and the the those the process used in exposure and the specific items reader during that we and would on the reviewing first those may have available industry, public. profiles; the the comment PUBLIC on We the in draft public welcome format development overall of process and the used in the include lay public risks originally of the associated planned document, like draw as to the candidates STATEMENT a public health with with the should be it to encourage for period. HEALTH to p r o v i d e the would strongly draft profiles as comment the profiles. the health development comments become general the the development. States, documents; human of Regis t e r notice, currently its of are rest are profiles them phases solicited We content summary the 12866) used COMMENT Federal the general -from all on of of on impact 25 eager A. The the first attention intended FR the m e t h o d s significant attention (52 PUBLIC previous substances. evaluating feedback development A and the of 1987 OF comment list are of levels close 17, professionals, comment public profiles. of hazardous are priority form, SOLICITATION should a statement concise chemical able to still / which statement of of concern. stand alone. be capable - 5043 of is the The If removed conveying 122 to the public substance. versions of different the We the public health general, other is The format for sections be used set subsequent sets - What this is presentation and number? Are would be LEVELS The process with each effect to used the significant to of the to "range" of not the such and content HUMAN this type selected that the can initial for of a information? graphic covers type both than of a upper single information that user? SIGNIFICANT the draft of health depending on may be while has effects data resulted gaps in circumstances essential in determining data gaps Toxicology data exposure suggestions m ay be (NTP) elements that This some less Program levels. for the associated individual critical process. DATA GAPS profiles other the N a t ional and define they EXPOSURE this the of to We modified of presenting identify the that audiences. might be have a way useful OF the the m o s t useful been made more range profile in this eventual comment in of values Is data gaps and we significant human requests toxicological format format levels. exposure, EPA, establish Our but the p r o f i l e s , g i v e n of presenting develop full identified exploring ways specifically way However, of ATSDR, the to p r o v i d e CERCLA different SIGNIFICANT a the to the chemical. evaluation effort has documents the IDENTIFICATION of needed on other ways levels been the of of Every of profiles reflects subset immediate. / of the documents? PROFILES effort (i)(3) by many OF useful generation E. identification format comment useful have these "k e y " , studies the v a l i d our best 104 timeframe. D. of in identified users profiles. there more Section of that of critical, or CONTENT OF THE including how lower bounds development important documents the m o s t first the represent to request the PROFILES chemicals. tight and of profiles, For capture FORMAT AND resource specifically and in the v a r i o u s as profiles of approaches. chemical-specific profile, required by clearly th e each that may be the draft on used profile of the a number comments for draft profiles constraints in the abbreviated evaluating invites been C. information statements alternative are of more IN THE DEVELOPMENT OF THE that we data toxicological and specifically effects associated with development statements notice for concerns the USED and studies concern here miss health DATA/STUDIES appropriate health considering recommendations B. In also f o r m a t s . This existing public solicits substantive are have are notice approaching this phase DEPARTMENT OF HEALTH 4 HUMAN SERVICES S P E C IA L Public Health Service Agoney lor Toxic Substances and Disease Registry Atlanta, GA 30333 FOURTH-CLASS RATE POSTAGE 4 F E E S PAID PHS/CDC Permit No. G 284 Official Business Penalty for Private Use $300 ABRAHAM W. HSIE DEPT.PREVENTIVE MEDICINE k COMMUNITY HEALTH DIV OF ENVIRON TOXICOLOGY UNIVERSITY OF TEXAS MEDICAL BRANCH GALVESTON TX 77550 '■ 5045 / ISP's/ \5 Y. 1 ^ 0 ^ V is it s i -177 5047 ’■ '^ & ’¥ • ; ■' : & -$T.. ; :j' ■• V ^ t ' 4 1 P - . ..... v;> 0» •• ■■,-■■.. ■ ■ - i ; ML. •V PRELE-UNAEY INFORMATION RELATIVE T O ___, •; THE TOXICITX-OF 2, 4-DICHL0R0PHEN0XY' , ACETIC ACID (E 2372 -3 ) - ..... \r-*.-• ; *•> •a •*,*vC‘w *♦ . • •*• • ■ : t ~V ^.. . — ; •• • - "" - rrVFfc T23. 14- 11-1 -'.&■. / : Or. I 219 A v .• ■;*; ■v.v w a 9-27-44 >- V raw 10-19-44 ^ W o A B j H. A. Wyse * w *»!* ■ w-.r . *. T* Technical Service & Deve?B±Jhent Div. (W. W. Allen) ♦.< • :f W. R. Veazey •• ', •-."Organic Research Laboratory ¿v- 'l' f ' ’*'» 1. • •’•# * « ? $ £ ... .. :v . * • • . ’ ...__ 1 ■ .. »■■■• • ;r-• £ v- V*‘ V :’ Ä ^ - 7 » Biochemical R a m é b k n t e jr THE DOW CHEMICAL COMPANY ***•«» k r S • .:v.. ~ V Rapt ■ pi. Br ' V - . /< . ¡ Z j. ^ : ■■ - ¿ 3 ■ - y y ".V-IV-**. J • ■#..'*'-*■*• -.v- •• •• «■•••••*»• ••,v- • : ;'f'/■ r ’-*-*- r^ - ; ‘v - .. ; \ v- Preliminary tests have indicated that 2^4-dichlorophenoxyacetlc acid has a moderate "acute oral toxicity and that " it does not produce marked skin irritation. •• .X.". •• iff:xj?'■ g .1. s . ix * A.' " We would not expect serious difficulties to result from J •■ . . • V * * ♦• « "• w *\* * •';''.*v-.Y' * •• * *’■“•’ 'T-’i handling this material in a reasonable manner. manner, 'v "*. * Similar tests vili be made on 2,4,5”trichloropheno^y- Ji / ■* • .' •j "'v acetic acid when a sample •■’- * ia-4 copies ** - ;r -, - . ' - .. Ti-f . '■-•r ' Î Y . f . . 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K e p h r r t t r'vice-.: t : r t t h e : n a i i u f a c t n r e : r e i r iic o u n u e nc d i f f i c u l t y i n t h o i r i ac t o r l - a • i c h . : v ..• /.lo y a c s , The e - ho r e e ra p lo y e a . i i i i n c c n t i n u . L '. e n t <.: n ct . c o u p e u n d i n e c n c o n t r i '- t e i fern»;:* F c r t:;;i.c .• :i.r», v.n-.. e r s t :. uc ,.hy i r c h o u l ä ::c t h a t :ac-n . h : : • •. ¿..yiln . ¿ r i e 1.li :i 1 i ì T • : . e i n f i e l c r-.:;.pivin“ t h e e h u a i e - . l s u b s ta n c G c i n a n i h i " : .a u l a cu. f a r • n y e f f e c t s vtòvorse t c Li 1 - * il.* J * *1 - . : e v .ertec i c n ■ c a c o i n Whie h a m. il i n vìi c.: c . • l i O: :i LI. . ... ,:a t e n e i v o l y v i *;li n cj -D ha a b ro k e n eu t 1. t ..e y a t t r i b u t e d t n i a c a s e t c o t n a : c . i, t e a t the e i v . p t i o u ; rii. sa ; . i. aravi i.u i ‘c «. 1 : c f .11 v. I t h f ; ,e u r a . e o f ■; e 1e; r r a ì n i n b i c V . . e*. T r a n c i ' t* ci f r -a: ir.,.»»* Jjv J o s e p h f . itcA in h cv fi y 7, 1 D4G » .* I <*,, a -ir.:; * ¿1-.. *"■. 1 1 I ¿1 ••, o O L * :v ” * J-* • • ’ > n r> / CCOCeli fr Z.7 J •J wi. « j V / X~ ^ #*f )« uoauai-j MN 0 7 3 2 8 0 C '- . - . C i T 0O »V O M 33 en ex 1 , V. :/0 *• •-J '-— !r^ 3 iyw3 Kja Vt • Ja 3, .►U -w* -J —'IwWW.—';.]• IwCÛ« t^aj “*) J .: .a j p -.jïi ->a ..j'3 •3. wie L: ;*»iculüU^.al Lab« "* i *J • * • ■- •*> *■* ‘ ^ V «.' >/ A o J L'W^'ciX ô O w —« U'« '. , r i l a r 3 . e t Ohio û ta to r . i l v o i ’Gi «y p rp o o n fco d : .• .• r e n e ? a ï t h o rcceirô c o a í t n p o t t h e . ..L . . v J 'C eh 'Z i'o l C o o t ï T e z e j » C h o 31 2 2 .3 r.:oo . j - . e j ' o t iloi-oloV èoûJ m -3 will be polished !X Î C ‘l".pe U a.;u '-ul - » « » e w e wU»--! c i b â v ü « - -ru! Very î/. ü . £. j r t a n t;cD/swP?- C V C 04G 13 5051 Za 5052 Za ✓ * J J ! 00 H2156896 Harrisson, Jos. V/. E., and Rees, Edward W. 2,4-D Toxicity-I. Toxicity Toward Certain Species of Pish. American Journal of Pharmacy 1 1 8 , 422-425 (1946). The L D 50 concentration of 2,4-D for minnows is approximately 2,000 p.p.m.; for sunfish, 1,000 p.p.m.; and for catfish, 2,000 p.p.m. The upper safe limit for minnows is 1,500 p.p.m.; sunfish, 500 p.p.m.; and esti­ mated to be about 500 p.p.m. for catfish. These figures refer to a seven-day exposure period during which the concentration of 2,4-D was kept at a fixed level. (R.H. Summary) 0012205 5053 3 5054 3 * DOW T &00264 l [ i f 1 ^ 3. Toxicity of 2 ,4-Dichlorophenoxyacetic Acid For Experimental Animals. Edwin V. Hill and Harold Carlisle. Journal of Industrial Hygiene and Toxicology. Vol. 29 pp. 85-95. March 1947. } V. b L L L 0002579 5055 DOW 500265 TOXICITY OF 2,4 DICHLOROPHENOXYACETIC ACID FOR EXPERIMENTAL ANIMALS* E d w in V. H il l 1 a n d H arold C arlisle * IT H IN the past few months several articles ( 1) have appeared in the scientific press describing the action and use of 2 ,4* dichlorophenozyacetic (2,4-D ) ad d and its am* monium or sodium salts as herbicides and plant hormones. This compound has also been used in orchards to prevent the premature dropping of apples. 2,4-Dicblorophenoxyacetic add is prepared commercially by reacting chloracetic acid, 2,4* dichlorophenol, and aqueous alkali or by the direct chlorination of phenoxyacetic add. It can be applied to plants either as an aqueous solution or dissolved in tributylphosphate diluted with diesel oil. .Potential health hazards may exist through inhalation when disseminated in aqueous or tributyl phosphate-diesel oil mists or by eating leafy vegetables or fruits contaminated by traces of the compound. Practically no data have been presented in the literature on the toxidty of this and related com­ pounds. Experiments were performed to deter­ mine the toxidty of the free add and its salts for ■experimental animals when administered orally, parenterally, and by inhalation. W M e t h o d s -a n d M a t e r ia l s Commerdal 2,4-D contains small amounts of phenolic impurities and related isomeric phe* noxyacctic adds. The following procedure was used to remove such impurities as well as inorganic contaminants and traces of colored material, a. Two hundred and twenty-one grams of 2,4-D were dissolved in 2 liters of 98-100 per cent ethanol with the aid of mild heating to increase rate of solution, b. T he resulting solution was suction filtered to remove small amounts of suspended matter which occasionally were present, c The filtered alcoholic solution was saturated with gaseous ammonia until a test portion • Received for publication August 1 1 ,’lWd. This work was conducted at Camp Detrick, Frederick, Maryland, from December, 1944 to June, 1945. ‘ LL Col., MC, AUS. •L l (Jg). USNR. of the clear supernatant liquid g a v e ' no further predpitate of the ammonium 2 , 4* dichlorophenoxyacctate. The solution was stirred continuously during the addition of the ammonia. d. The predpitated ammonium salt was suction' filtered, dissolved in 2 liters of hot distill««* water (about 70* C.), and 10 grams of de­ colorizing char added to the ammoniacal solution, which was then allowed to ««»«A at 70* C. for one-half hour. e. The decolorizing char was removed by gravity filtration and the filtrate cooled at 10* C. The ammonium 2 ,4-dichlorophenoxyacctatc gradually crystallized in fine needles, the crystallization being complete after twelve hours. f. The ammonium salt was suction filtered, dissolved in 2 liters of hot water (70* C.) and the 2,4-dichlorophcnoxyacctic add pre­ dpitated by the addition of the alkaline solution, with stirring and cooling, to ISO ml. of concentrated hydrochloric add (38 per cent S.G. 1.19). g. The 2,4-dichlorophcnoxyacetic add was suction filtered, washed with distilled water until a test portion of the filtrate gave no predpitate or cloudiness with aqueous silver nitrate, and re-crystallized three times from hot water. h. The filtered product, vacuum dried over concentrated sulfuric add for seven (lays, consists of fine transparent necdldike crystals, m.p. 139* C. (A Fisher-Johns melting point apparatus was used for the determination), reported m.p. (literature) 138* C , 139-140* C The neutralization equivalent was determined for 3 samples, 1) 221.1, 2) 221.1, 3) 221.0. Toxidty determinations were made on crude and purified 2,4-D . Sodium and ammonium salts of the purified acid were prepared and also assayed for toxidty at concentrations of 1 or 4 per cent. Solutions of the acid or salt were pre­ pared in jihysiological saline and adjusted to a final pH of 7.0 to 7.2. In the parenteral toxidty tests on these four materials, physiological saline o m o. 5 ^ "V I Z 9 o m a z I I eI M • i i I l i i 5056 0 0 0 2 6 8 0 86 JOURNAL OF INDUSTRIAL HYGIENE AND TOXICOLOGY r R esu lts Tie Acute Tosieiiy of tUt Purified Sodium Sail of 2,4-D A total of 17 experiments was carried out to determine the toxicity of the purified sodium salt of 2,4-D for the various experimental animals when given by injection or feeding with a stomach tube. In these experiments approximately 450 mice, 150 white rats, 125 guinea pigs, 70 rabbits and 3 monkeys were used. Since the titrations proved to be more precise and repeatable than titrations involving living agents such os bacteria, it was found possible to arrive at a fairly accurate LD m by using only 10 mice, 6 rats, 6 guinea pigs, and 4 rabbits for each 4 or 5 dilutions, covering a comparatively small range o f concentrations. For example, the mouse titrations consisted of tho injection or feeding of 2.5 mg., 5.0 mg., 7.5 mg., and 10- mg. of 2,4-D . The amounts usually administered to_guinea pigs were 100 mg., 150 mg., 2C0 mg., 250 mg., and 300 mg. Most of the titrations were earned out at lc.'-'t twice and in every replicates agreed satit.fr.cicr;!/. The toxic and tolerated doses determined b y these 17 titrations are given in .Table 1. Tiro LDi» of 2,4-D injected intravenously in rabbits is approximately 400 mg./kg. About 50 per cent of the animals die suddenly of acute ventricular fibrillation and those that survive this acute episode die within several hours to three days. The outstanding symptoms were referable to the skeletal muscle where a condition not unlike myotonia congcntia was present. The animals received symptomatic relief by the administration of quinine hydrochloride, but a fatal outcome could not be avoided. The symptoms observed in the smaller animals were, in order of appearance, stiffness of the extremities with some muscular incoordination, lethargy, paralysis of the hindquarters, stupor, coma, and finally, death. These symptoms were observed consistently regardless of the method of administration of the chemical. The data indicated that the toxic and tolerated doses are slightly higher when the material is given by stomach tube. As would be expected, animals fed with 2,4-D did not develop symptoms as early as did injected animals. In addition, symptoms were more variable in degree in animals fed by stomach tube and the 'final results of such titrations were less precise and repeatable. Table 2 summarizes the results of one of several ex­ periments carried out in which comparative data on toxicity by feeding and injection were obtained. Three monkeys were fed varying amounts of the material. One monkey receiving 0.75 gm. exhibited only slight atypical symptoms and re­ covered completely. Two additional monkeys were fed 1.0 gm. and 1.5 gm. each, but both p-mW.!« became ill about four hours after lyin g fed and regurgitated a large portion of the material so that no information on lethal doses for monkeys by feeding is available. N o typical sym ptom s' appeared in any of the monkeys, and all were apparently normal seven days after being fed. One animal, which was fed 1.0 gnuof 2 ,4-D ,w as kept under observation for an addlr^n-t e i^ y Dov^ 500266 was ordinarily used as a volume control but in later experiments a 1.81 per cent sodium chloride solution was used as an osmotic pressure and sodium ion control. The preparation of oil solutions of crude-acid required the use of an intermediate solvent. The acid was fust dissolved in either tributyl phosphate or n-butyl alcohol and these solutions were then mixed with the proper amount of diesel oil. Var­ ious combinations of 2,4-D , solvent and 0 2 diesel oil were tested. Each combination tested will be described in detail as the experimental work on the material is presented. All injections were made by accepted technics. Forced feedings were carried out in the smaller animals by means of a 04 or $S woven doth ureteral catheter and in the larger animals by means of a 08 rubber ureteral catheter. The catheters were attadiud to syringes of appropriate size and were lubricated slightly with petrolatum before use. It was found advantageous to an­ esthetize white rats and guinea pigs before feeding was carried out. Since nothing was known about the length of time necessary for effects to occur, all animals were held for more than an adequate number of days during the fust part of the work. It was found that deaths seldom occurred after the fifth day so that in the later experiments animals were sacrificed on the sixth or seventh day. days without the appearance of any delayed efforts. Two of these same monkeys were then used in injection experiments. The monkey which had been fed 0.75'gm . was injected mtrapcritoncally with 1.0 gm. of the purified sodium salt of 2 ,4 -D . U rn aaimal developed nausea and v o m itin g 5057 0 0 0 2 G51 TOXICITY OF 2.4 DICHLOROriIEXOXYACETIC ACID stillness o(.ihc less with some muscular incoordina­ tion, lethargy, hanging of the head and ptosis of the eyelids. The symptoms appeared about two 87 which had been fed 1.5 gm. seven days previously, was injected intraperitoncally with 1.5 gm. of the same material, l i t i s animal showed symptoms TABLE l Toxic and “ ole sated D oses o r tbz Sodiuu Salt o r 2.4-D ro a ExrESUUEVTAL An iu a u av I naction and F eeding AffafflUUATS VSIC3T 07 AMttlAl Per Asiaci Bx-Af. Per Aaiaul ms-Ae. White mouse..................... Intraperitoneal White mouse..................... Stomach tube 20 gm. 20 gm. 7.5 mg. 7.5 mg. 375 375 2.5 mg. 2.5 mg. 125 125 Guinea pig......................... Intra|>eritoneal Guinea pig........................ Stomach tube 300 gm. 300 gm. Rabbit................................ Intraperitoneal Rabbit................................ Stomach tube Rabbit................................ Intravenous 2.5 kg. 2.5 kg. 2.5 kg. n n o o or AMiMt* tïAtlOM IROU White rat.......................... White ra t.......................... Intraperitoneal Stomach tube 150 gm. 150 gm. Monkey.............................. Intraperitoneal Monkey.............................. Stomach tulie 3.5 kg. 3.5 kg. TOXICDOCS* 209 mg. 300 mg. 1.0 gm. 2.0 gm. 1.0 gm. ICO mg. 100 mg. TOtXSATXDDOSS** 666 1000 100 mg. 100 mg. 333 333 400 800 400 500 mg. 500 mg. 500 mg. 200 200 200 666 666 25 mg. 50 mg. 166 333 1.5 gm. 750 mg. 428 214 ¿9200QM o a orci JW7\ * Toxic dose—50 per cent mortality. ** Tolerated dose—largest amount causing no deaths. TABLE 2 Cou? a2ison o r tbs T oxioty o r tux Pusitied Çodxdu Salt o r 2,4-DiarLoaornENOXYACETic Acid roa White R ats bv I njection and F eeding HO1,4-0 OlVXM soon AfPCASANa or SVUROHS MOBTA&» 50 ICO 150 i.p. i.p. i.p . LP. Feed Feed Feed Feed LP. Feed 1 hour 30 min. 30 min. 30 min. 4-6 hours 2-3 hours 1 -2 hours 1 -2 hours None None 2/6 6/6 6/6 6/6 0/6 200 50 ICO 150 200 Saline control Saline control in* 4/6 5/6 6/6 0/6 0/6 * Mortality data recorded as ratio of deaths to total animals used. Rats used in this experiment averaged 150 gm. in weight. hours after inoculation, persisted for about fortyeight hours, and then began to lessen in intensity. A t the end of five days, the animal had recovered and was apparently normal. The second monkey, similar to those seen in the fust injected monkey and by five days had recovered completely. Toxicity of Crude Venus Purified 2,4-D Five titrations were carried out to com|>arc this toxicity of purified sodium salt of 2,4-1) with that of the crude material as it comes from the manu­ facturer. In these cx]>crimcnt<, the crude add was first converted to the sodium salt and was used in this form with all impurities present. The re­ sults, as presented in Table 3, indicate that there was no significant difference in the toxidty of the two materials cither by injection or feeding. The Toxicity of Pure 2,4-D Two experiments were carried out to test the toxidty of pure 2,4-D . Because of its insolubility, the material was suspended in S per cent gelatin solution and fed to white rats by means of a stomach tube. The addition of gelatin to the suspension prevented settling ami agglomeration of the partidcs. N o injection experiments were carried out, but the results of the two feeding titrations summarized in Table 4 indicated that 2,4-D possesses about the same toxidty as does its sodium salt. 5058 0002682 -j JO UHMAL OF INDUSTRIAL HYGIENE AND TOXICOLOGY TASLE 3 « f n —.. - rrn-f n- t~ Tnvrnrv n? PuiLTOD ANDCiUDE SODIO!! SAIT 07 2,4-D 3Y IirjTCROtt Atm 2 Y * — - - 3 o o uoatAUX»*** so n s White mouse SCO 375 250 125 Saline control I.P.* I.P. I.P. I.?. I.P. 1CG0 333 167 Saline control I.P. LP. I.P. I.P. I.P. 833 667 5G0 33 Saline control I.P. I.P. I.P. I.P. I.P. 4C0 2C0 Soline control I.P. I.P. I.P. 3/3 0/3 1C00 Feed** Feed Feed Feed 5/6 3/6 White rat 666 Guinea pig * Rabbit White rat 666 333 Saline control . Parì£id Crai* 10 /10 10/10 3/10 4/10 0 /10 0 /10 0/10 0 /10 CoEtrsla 500268 KO./ZO. irscca 0 /10 6/6 5/6 5/6 4/6 0/6 0/6 0/6 0/6 6/6 5/6 4/6 4/6 1/6 0/6 2/6 0/6 0/6 0/6 1/3 0/3 0/3 1/6 5/6 2/6 1/6 0 /6 * LP.—Intr&peritoneal injection of 2,4-D salts in physiological saline solution. ** Feed—Feeding by stomach tube of 2,4-D salts in physiological saline solution. *** Mortality data are recorded as ratio of deaths to total animals used. T A 2L E 4 A Coumusoh or m T oxicity o r P u u 2,4-D and tsx Ptnunxn Sonimi Salt o r 2,4-D by F ekoimo to White Rats and G uinea Pics U O ITAU TV* in a u moJ e o . 1 .4 -0 White rat 1CC0 ■ 666 333 Gelatin control Soline control Guinea pig 33 Celatin control Soline control 4/4 3/4 0/4 S o d iti» S a ilo ( 2 .4 * 0 C o» tr» U 5/6 3/6 0/6 0/4 0/6 0/8 0/8 0/4 0/4 * Mortality data recorded as. ratio of deaths to total animals used. The Toxicity of the Ammonium Sait of 2,4-D Experiments were carried out to compare the toxicity of the ammonium salt of 2 ,4 -D with that of the sodium salt. Amounts of the ammonium salt equivalent to the tolerated dose of the sodium salt for each species were fed to groups of 6 guinea pigs, 6 white rats, and 4 rabbits. l a this experi­ ment all animals survived, indicating that the ammonium salt is a t least no more toxic than the sodium salt by feeding. Two injection titrations were carried out on white mice and the results indicated that the ammonium and sodium salts possess about the some degree of toxicity by in­ jection also. The precoding two types of experi­ ments were carried out with both the purified salt and the crude salt and no difference in to x id ty was detected. One monkey was fed 1.0 gm. of the pure ammonium salt of 2 t4-D. T h e monkey regurgitated a large‘amount of the material about 000268 ¿ fo r th IH 7 ) TOXICITY OF 2,4 DICXLOROPHENOXYACETIC ACID The Toxicity of Crude Acid 2,4-D Dissolved in either Tributylphosphate or n-Buiyl Alcohol and Oil The toxidty of crude add 2,4-D dissolved in tributylphosphate and oil was investigated to tributylphosphate in oil, and a feeding titration in guinea pigs using 4 per cent 2,4-D and 8 per cent tributylphosphate in oiL It can be seen from Table 6, which summarizes the results of the first two experiments, that when a 1 per cent solution was used deaths occurred at one-fifth the tolerated dose of 2 ,4-D and all control animals died, whereas, when a 4 per cent solution was used, only occasional deaths occurred in the control group and in the scries of animals fed the tolerated dose. In view of the results of the first two experiments it was DOVV500269 three hours later and was apparently normal when observed the next day. N o symptoms other than the nausea were noticed. 'The results of the experiments on the ammonium salt of 2 ,4 -D arc outlined in Table 5. 89 TABLE 5 A Comparison o r tax Toxzcmr or Poax aks Cam» Annotami Salt with tmx Pu is Sooiun Salt or 2,4-D u o t i A llT T * * * sn a zs K S -/3 0 . BOOTS A m m o n iu m S a l t Pvt White mouse «Vhite rat Guinea pig - Rabbit Monkey C ru d o S o d iu m S a l t P u ra 7/10 3/10 10/10 4/10 0 /10 0 /10 1 /1 0 0 /10 0 /10 0 /10 9/10 500 37S 250 125 Saline control I.P.» I.P. i.p. i .p . I.P. 333 Saline control Feed»* Feed 33 Saline control Feed Feed 200 Feed Feed 0/4 Saline control 285 Feed 0 /1 C o a tro la 3/10 0/10 0/6 0/6 0/6 0/6 0/6 0/6 0/6 0/4 0/4 * I.P.—In tra peritoneal injection of 2,4-D salts in physiological saline solution. ** Feed—Feeding by stomach tube of 2,4-D salts in physiological saline solution. *** Mortality data are recorded as ratio of deaths to total animals used. determine whether or not the combination of solute and solvents was significantly more toxic than the solute alone. It was not the purpose of this investigation to determine the toxic levels of the solvent. One gram of 2,4-D is soluble in 2 ml. of tri­ butylphosphate and the resulting solution is miscible in all proportions in oiL Various con­ centrations of the agent in tributylphosphate and oil were prepared by altering only the amount of oil used and leaving the agent-solvent ratio con­ stant. The first two experiments on the 2,4-D tributylphosphate oil complex were preliminary in taturc and consisted of a feeding titration in white rats using 1 per cent 2,4-D and 2 per cent decided to test a constant amount of 2,4-D (the tolerated dose) in various amounts of tributyl­ phosphate and oiL For this work, concentrations of 1 per cent, 2 per cent and 4 per cent 2,4-D in oil containing 2 per cent, 4 per cent, and 8 per cent tributylphosphate respectively, were used for feeding experiments with guinea pigs, white rats and rabbits. When a 4 per cent solution of tributylphosphate in oil containing a tolerated dose of 2,4-D was fed to experimental animals in a comparatively small volume of oil, all or most of the animals survived. Only occasional deaths were observed in the groups of guinea pigs and white rats, and all of the rabbits survived. When the 2 per cent solution was used, b 0002684 JOURNAL OF INDUSTRIAL HYGIENE AND TOXICOLOGY so an amount of 4 per cent 2,4-D in oil containing 0.5 grains 2 ,4-D . This volume of material, 12.5 CC-, caused very violent regurgitation soon after the feeding was carried out, but the animal was apparently normal the next day. N o symptoms other than the nausea were noticed. Three experiments were carried out to test the toxicity of n-butyl alcohol solutions of 2 ,4-D . These experiments were similar in nature to those carried out with the tributylphosphate-oil solu­ tions. Two groups of rabbits were fed the toler­ ated dose of 2,4-D (0.5 gram) in 2 per cent and 4 per cent solution and all animals survived. In DOW 500270 A-cr cent tributylphosphatc in § 2 diesel oil. The amount of this material tolerated by the experi­ mental animals was equivalent to aliout 200 cc. for the average sized man. One monkey was fed comparable experiments in guinea pigs and white rats, only an occasional death occurred. Occa­ sional deaths were also observed in the control groups which received comparable volumes of n-butyl alcohol-oil mixture. Although not con­ clusive, these experiments indicated that n-butyl alcohol oil solutions of 2,4-D are at least of the same order of toxicity os the tributylphosphatc oil solutions. Sub-acute Intoxication in Dots A group of 8 dogs received 2 ,4 -D intravenously, varying from two injections of 200 m g./kg. each to six injections of 25 m g./kg., the injections being given at daily intervals. Two dogs (nos. 1 and 2) receiving two injections , each of JOQ m g./kg. died on the third and second 5061 0002655 TOXICITY OF 2,4 DICHLOROPHENOXYACETIC ACID day respectively after the last injection. Tne first symptom was an unsteady gait, with the extremities somewhat stiff and extended. On the day following injection both animals were lethargic and stood up only when forced. The gait was still unsteady and extremities stiff. The deep reflexes of both animals were equal and active, pupils in mid dilation and equal. Corneal reflex in dog no. 1 was absent in contrast with dog no. 2. Nictitating membrane was sluggish in dog no. 1 covering one-half of the eye I¿all; it was active and in normal position in dog no. 2. Two dogs (nos. 3 and 4) received two injections of 200 m g.A g. each on successive days. Both animals died on the second day. They were semi-stuporous for the first day after injection and responded slowly to stimulation. Pupils did not react to light and corneal reflex was either absent or extremely sluggish. Deep reflexes were equal and active in dog no. 3. The polymorpho-nudcar leucocyte count was 13,030 before injection and 1,400 the day following injection. Similarly, the lymphocyte count decreased from 3800 to 8C0. D og no. 4 showed no significant change in the number of polys but did show a reduction of ymphocytcs from 5,CC0 to 2,200. One dog (no. 6) was given six injections at daily intervals of 25 m g.A g. of 2,4-D . On the second day this animal showed blood oozing from the gums at the tooth margins which did hot increase in severity. On the fifth day after injections were terminated, decubitus ulcers developed over the outer aspect of the forelegs and right thigh which gradually became more extensive, deeper and infected. Neutropenia or lymphopenia were not apparent at any tme during the illness. The animal was sacrificed on the ninth day. Two animals (not. 7 and 8) received six injections, a t daily intervals, of 50 m g./kg. each. D og no. 3 died on the third day. This animal became lethargic and semi-stuporous after the third injec­ tion. On the second day after the series of injec­ tions were terminated conjunctivitis developed. Blood oozed from the gums and tooth margins, the gait became unsteady and coarse muscle tremors appeared in the extremities after moderate exercise. The deep reflexes were equal and active. The rectal sphincter was lax and allowed the escape of light, watery brown stools. The animal became more lethargic on the third day and died in the mid-afternoon. Polymorpho-nudcar leucocyte count decreased from 6,CG0 to 2,500 and 91 the lymphocyte count from 3,500 to 200 as a result of the intoxication. The platelet count, originally 323,000, dropixai to 138,000 in the post-injection .period. Dog. no. 7 dcvclcri necrosis of the gums on the right and left side of the lower jaw along the posterior as|)cct where the teeth of the upper jaw overlap those of the lower jaw. Blood oozed from the gum margins throughout the remainder of the mouth. The necrotic process gradually extended to involve the floor of the mouth, the buccal mucous membrane and exposed much of the mandibular bone. Decubitus ulcers developed on the outer aspect of the extremities on the Kth day. The animal was sacrificed on the sixteenth day when rectal temperature was elevated for the first time. This animal did not show any significant reduction in the polymorpho-nudcar count. There was, however, a reduction in the lymphocyte count from 2,600 to 1,100. It was interesting to note that smaller doses of 2 ,4-D appeared to have a cumulative action. The development of the decubitus ulcers and severe necrotizing lesions of the mouth occurred in all animals receiving 2,4-D over a protracted period. In general, the animals showed a slight reduction in the red count, hemoglobin level, and an increase in the plasma non-protein and urea nitrogen. There was no significant change in the total protein content of the plasma. At no time did any of the animals show any signs of icterus. DOVV500271 -Arch 1M7] Sub-acult Oral Toxicity in Rats Young male rats, weighing around ICO gnu. each, were put in individual metabolism cages and fed a stock diet of sucrose, Labco casein, Crisco, and salts. In addition they received a daily vitamin supplement containing pyridoxine, ribo flavin, nicotinamide, calcium pantothenate, choline chloride and thiamine. Following an observation period of one week, the animals were divided into 4 groups of 7 mts each. Croup 1 received 100 mg. of 2,4-D /kg. of diet; Group 2, 200 mg.; Group 3, 400 mg.; Group 4 served as controls. N o difference could be observed In the food consumption and growth rate of animals receiving 100 and 200 mg. of 2,4-D /k g. of diet. One animal in the group receiving 500 mg. of 2,4-D of diet died on the tenth experimental day, but In as much as no other member of the group was affected, the death was considered incidental. In view of the negative nature of the experiment, the diet of the rats of group 1 was changed on t h e 00025S JOURNAL OF INDUSTRIAL HYGIZNE A N D TOXICOLOGY twenty-first experimental day to contain 1CC0 mg. of 2,4-D /k g. of diet. Despite this large dose there was no evidence of reduced food intake and co effect on the growth rate during the subsequent two weeks. The experiment was discontinued one month from the time 2,4-D was first intro­ duced into the diet. It was concluded that the rats* diet containing as much as 0.1 per cent 2 ,4-D was not intoxicating. Sub-acute Oral Toxicity in Guinea Pits Two groups of 6 guinea pigs each were fed daily 50 mg. and ICO mg. respectively of the purified sodium salt of 2,4-D . On two occasions it was impossible to carry out the feedings on schedule so that the animals actually received only ten feedings in twelve days. Five out of 6 of the guinea pigs receiving 50 mg. at each feeding sur­ vived. These 5 animals were fed a total of 0.5 gm. in twelve days. Three out of 6 of the guinea pigs receiving 100 mg. at each feeding survived. The total dosage of tins group was ID gm. in twelve days. These results are nullified to a certain extent by the fact that three deaths occurred in the saline controls, making it appear that all deaths might have been non-specific and caused solely by the rather rough treatment associated with daily feeding by stomach tube. Supporting this con­ tention is the fact that none of the animals showed the typical complete paralysis before death. Aside from these discrepancies, the fact remains that 3 guinea pigs survived a total of 1.0 gm. of 2 ,4-D , .which is about 3 or 4 times the amount necessary to kill when given at one feeding. The Toxicity of the Sodium SaUof 2,4-D by Inhalation Attempts were made to produce symptoms in guinea pigs by exposing them to a wet cloud of sodium salt of 2,4-D in a spray chamber produced by nebulizing an aqueous solution of the salt and to a dry cloud of the crude add produced by directing a jet of air against a finely pulverized dry powder in the bottom of a small chamber. Tn no case did any of the animals exhibit typical symptoms and there was no gross evidence of lung irritation, even though concentrations of the material in the air were high and exposure times were tong. In the first experiment carried out in the spray chamber the animals were exposed to a CT (mg. min. per M 1) of approximately 6000-8000. The second experiment was not quantitative in that it was --i WSOOS Moa 92 impossible to calculate the CT given. I t hon been estimated that the animals were expored to & C T even greater than used in the spray chamber. N o data was obtained on the average ciac of the particles in the aerosols or thè fraction of the material impinged out of the inspiratory air and absorbed by the animai. Pathelopeol Chanfes Rats and guinea pigs dying of massive doses of 2,4-D were sacrificed sixty to ninety-six hours after administration of the chemical were autopsied (the central nervous system was not examined). Constant findings on gross examination were con­ gestion of the viscera and enlarged swollen kidneys. On section of the kidneys, the parenchyma bulged from the cut surface and the capsule stripped almost spontaneously presenting a dull yellowishred surface. The cortex was swollen and poorly differentiated from the medulla. Microscopic examination of the kidney revealed the swollen cortex to be due almost entirely to a massive clouding swelling of the epithelium of the proximal convoluted tubules, which in many cases com ­ pletely occluded the lumina. Some of the cell membrane had ruptured, with the escape o f the cytoplasm and nuclei to form an acidophilic debris in the tubules which was latex converted to hyaline and cellular casts found in the collecting tubules. The glomeruli and blood vessels were unaffected. Occasional animals showed slight patchy pulmonary edema and alveolar hemor­ rhages. The livers did not show any significant pathological changes. A group of 6 rabbits receiving from four to thirteen daily (except Sunday) injections of 50 mg./kg. were sacrificed on the day of or the day following the last injection. N o striking patho­ logical changes were observed in this group of rabbits. The kidneys of all animal« but one showed slight to moderate degenerative changes consisting of granular or vacuolar swelling of the cells of the proximal convoluted tubules. T he glomeruli were unaffected. Stimulation of hema­ topoietic and rcticulo-endothelial tissues was suggested by the occurrence of hyperplasia of the lymph nodes and spleen in several animals. Mod­ erate hyperplasia of the bone marrow associated with the extra-medullary hematopoiesis in the liver, spleen, sa d adrenals. This effect was not consistent throughout the group. Significant blood destruction, evidenced by considerable 0002SG? ( TOXICITY OF 2,4 DICHLOROPHENOXYACETIC ACID hemosiderosis of the 'spicvn, was present in only one animal. A review of the biood hemoglobin levels in this animal showed a decrease in the hemoglobin from a pre-injection level of 9.0- 9.7 gm. to 7.8 gm. after five injections. Of three other rabbits in this group receiving as many or more injections, two showed no significant change in hemoglobin level and one showed a slight decrease. The liver showed no significant changes. The lungs showed scattered patcchial hemorrhages in a few eases and moderate patchy edema in two. Another group of 6 rabbits receiving from three to seven daily injections of 100 mg./kg. were sacrificed on the day of or the day following the last injection. There was a significant decrease in hemoglobin and red ceils in one animal of this group, and a moderate decrease in hemoglobin in three others. N o changes in the white cell count or differential count were observed. The kidneys of this group of animals also showed parenchymatous degeneration as the most constant change. Marked hyperplasia of the bone marrow, associa­ ted with extra medullary hematopoiesis, occurred ' i one animal. Lymphoid and splenic hyperplasia ere observed in several animals but was not a constant finding. Hemosiderosis of the spleen was observed only once, in the animal showing the considerable decrease in hemoglobin anrl red cells. Evidence of slight lymphoid destruction was ob­ served in the spleens of two animals of this group. The liver showed no significant changes. Dogs succumbing to massive doses of 2,4-D or sacrificed were autopsied. In contrast with the other species studied, dogs showed a considerable susceptibility to the development of liver damage. The hepatic lesions consisted of ccntro-lobular degeneration, atrophy and lysis of the parenchy­ mal cells about central veins of the lobules, with congestion and dilatation of the pericentral sinu­ soids. In one dog, dying at thirty houn after two injections of a relatively high dose (200 mg./kg-)i there was an extensive though early necrobiosis throughout the liver. The kidneys showed cloudy swelling of the tubular epithelium; the reversible nature of this lesion is indicated by its absence in animals sacri­ ficed or dying after some delay after the last injection. On the other hand, one dog receiving a large dose (4C0 mg./kg-) showed actual tubular epithelial necrosis, similar in type to that observed in mercury poisoning. Striking lymphoid necrosis the lymph nodes, thymus and spleen was 93 evident, especially at the higher doses. Unequivo­ cal bone marrow changes were not observed, although it seems quite possible that with the proper dosages of the chemical such lesions might occur. An occasional animal showed evidence of blood pigment deposition in the spleen and lymph nodes. Minor focal parenchymal lesions were observed in the adrenal in several instances, accomitanicd by infiltration of the polymorpho­ nuclear leucocytes. DOW 500273 i f arch 19J7\ D iscussion The investigations disclosed that 2,4-D is a relatively non-toxic compound for experimental animals, having an LDt*, expressed in mg./kg., of 375 for mice, 1000 for guinea pigs, 666 for rats and 800 for rabbits when administered orally in aque­ ous solution. All of these species reacted similarly to the chemical. There was no apparent differ­ ence in the toxicity between crude acid and a highly purified preparation, or between the sodium and ammonium salts of the acid. Monkeys were shown to be able to tolerate 428 mg./kg. of 2,4-D when administered intraperitoncally. However, when the material was given in large doses by mouth, 1 or 1 ) grams, the animal became nauseated and vomited a large portion of the material so that accurate information as to the toxicity of the compound administered by the oral route in this species is lacking. Three-fourths of a gram of the material was given to one monkey without the development of vomiting or serious illness. Based on the best data available, which arc recognized to be inadequate, monkeys can tolerate single dosages equivalent to 214 mg./kg. In any assessment of the acute toxicity of a chemical based on data obtained from laboratory animals it should be borne in mind that considera­ ble variation in species susceptibility may occur and that the data obtained cannot always be translated into the toxic doses for humans. In this case, however, all of the laboratory animals tested reacted in a similar fashion to the material so far as could be determined from signs and symptoms which developed and from the patho­ logical lesions which were present at autopsy. Assuming that man is no more resistant or suscep­ tible than the rabbit or monkey, then the largest tolerated dose for a 75 kg. man would be 15 gras. With the exception of the monkey, all of the laboratory animals used lacked the vomiting" reflex to that they were unable to relieve them- 0002S38 I. JOUIIXAL OF INDUSTRIAL HYGIENE AND TOXICOLOGY f ‘I 1 ï f J1 I 1 b ! j- L L l i doses over a period of twelve day3 without any harmful effects. The experiments to determine the toxicity of the sodium salt of 2,4-D by inhalation tend'd to indicate that the material was relatively non-toxic when wet or dry clouds were inhaled. It should be pointed out that in none of these experiments was the average particle size of the aerosol determined nor the amount of material retained and absorbed by the respiratory tract determined. Further studies on the toxicity of this compound by the respiratory portal of entry will be needed before any final conclusions can be made. In view of the fact that the material did not produce any evidence of lung irritation and that the toxic oral dose of the material for guinea pigs is relatively high, it would tend to indicate that the material would be relatively non-toxic by this route. 2 ,4 -D dissolved in a solvent complex of tri­ butyl phosphate and diesel oil has been recom­ mended for use as a herbicide. This particular form , of material has the advantage over the aqueous solution in that the tributylphosphate acts as a co-agent or synergist to the 2,4-D producing greater plant damage than could be accounted for by the acid alone. T he present study did not attempt to determine the toxic or tolerated dose of tributylphosphate or diesel oil. There was no evidence of a synergistic or addi­ tive effect when a tolerated amount of 2 ,4 -D was dissolved in the tolerated dose of tributylphosphate oil complex and administered to the experimental animal*. Production of chronic poisoning by this material was not attempted. o o 500274 I ïd v c3 oí irritating material by vomiting. The experiments conducted ia monkeys indicate that the material is a gastric irritant in large doses, so that the possibility of the occurrence of acute poisoning in humans would seem relatively remote because of the large dose which man could pre­ sumably tolerate. Assuming that man is no more susceptible than the most susceptible animal tested, the mouse, then the calculated oral LDi* for man would amount to approximately 28 gnu. Sub-acute intoxication was produced in dogs by giving daily injections of the material over a period of six da)?. With the administration of smaller doses, 25 mg./kg., there was a suggestion that the material had a cumulative anion. In contrast with the other species studied, dogs showed a considerable susceptibility to the development of liver damage. Occasional lymphoid necrosis in the lymph nodes, thymus, and spleen occurred especially with high doses. A few animals demon­ strated a significant reduction in the number of lymphocytes in the circulating blood. The de­ velopment of decubitus ulcers and their occasional association with a significant reduction in tho leucocyte count was a disturbing observation. Reduction in the leucocyte count did not occur in every ease, but its occasional development should direct attention to blood studies in cases of sus­ pected intoxication in man. Investigations on the sub-acute toxicity of 2 ,4-D by the oral route were limited to two species, rats and guinea pigs. Rats were fed varying amounts of the material up to ^ per cent by weight of their diet for á period of one month without any significant effect on their food intake, rate of growth, or the development of any charac­ teristic signs of intoxication. Guinea pigs, which were fed 100 mg. per day of the material by stom­ ach tube over a period of twelve days until a total of 1 gram of the material had been administered, did not develop characteristic evidence of intoxica­ tion. In this particular experiment, non-specific deaths occurred in approximately the same per­ centage in both the test and control animals. All of these deaths were believed to result from trauma, associated with the passage of a stomach tube a t frequent intervals, in as much as none of the test animals developed the typical paralyses or skeletal muscular signs observed in poisoned animals. It was therefore concluded that guinea pigs could tolerare 1 gram of 2,4-D in divided S l’MUARY The LD m expressed in m g./kg. of 2,4-dichlorophenoxyacetic acid by mouth is 375 for mice, 666 for rats, 800 for rabbits, and 1000 for guinea pigs. The tolerated dose for these same species was 125, 166, 200 and 333 m g./kg. respectively. The largest dose administered to monkeys without ierious after effects was 214 mg./kg. Subacute intoxication waa produced in dogs by the daily administration of 25 m g./kg. over a six day period with the development of liver damage. Lcucopcnia was observed in a few »nimnU. Rats were fed up to per cent by weight of their diet over a period of one month without any harmful effects. Guinea pigs can apparently tolerate 1 gram of the material administered in divided doses of 100 mg. over a period of twelve days. 5065 0 0 0 2 5 2 9 lÎ6 r tk lW \ TOXICITY OF 2,4 DICHLOROPUENOXYACETIC ACID AacMowuDOZunrr The authon wish to thank Major Arthur Glazier and Captain Morton Golds ton of the Medical Division, Edgewood Arsenal for permission to use their data on subacute toxicity of 2,4-D for dogs and chronic toxicity in rats. 500275 A limited amount of work on the toxiaty of wet and dry clouds of the sodium salt indicated that it was relatively non-toxic. The toxicity of 2,4-D dissolved in tributylphos* phate and diesel oil was not enhanced by 'these solvents. BIBLIOGRAPHY (1) HanzBOAMs, E. M.: War on weeds. Science, 103: 465, 1946. ’ S p e c ia l P o o le r s DivistOM, Csmncai W a x p a u Szavicz: Plant growth regulators. Science, 103: 469, 1946. VAM OVTSSIEX, J . AMD VXLXZ, ISUAXLt Use of 2,4-dlchlorophenoxyacetic acid as a selective herbicide in the tropics. Science, 103:- 472, 1946. S io t h , F. G., H am m , C. L. a m d C a o u o m , R. F.: Control of ragweed pollen production with 2,4- dichlorophenoxyacetic add. Sdencc, 103: 473, 1946. H iuzsbamd, E. M.i Herbiddol scU'oa of 2,4dichlorophenoxyacetic sdd on the water hya­ cinth, eirhomia aassipca. Science, 103: 477, 1946. Emus, W. B., T uoupsom, H. E. and S u m , H. H.t Tributyl phosphate at a solvent for preparing concentrated and oil-miscible solution* of 2,4= oichlorophenoxyacetlc add and similar sub­ stances. Science, 103:476,1946. 5 0 6 6 0002630 5067 i - \ BIOCHEMICAL RESEARCH LABORATOR? THE DOW CHEMICAL COMPANY Pile T23.14-11-1? Chg. 1219 Rec'd Pin'd 6-9-48 Work B y E.M.Adams Subject TOXICOLOGY AND HYGIENE: --------- 2,4-DICHLOROPHENOXY ACETIC" ACID To: Britton's Div. Att: B. N. Schrauf R. C. Dosser J. B. Arnold Safety Dept. Att: John Knight Dr. H. H. Gay T2.2-10-1 t Rept. By ch ef k é-Zc '1 ? UNIT INDEX cn A summary of available information. cn INDEX HEADINGS Acetic acid, 2,4-dichlorophenoxy SUMMARY OF HAZARD The toxicity is such as to present but minor hazards of systemic effects. Skin irritation is possible from heavy prolonged contact with strong solutions or the solid acid. There is no hazard _1 _ of absorption through the skin. SUMMARY OP TOXICITY The systemic toxicity was moderately high as Indicated by single and repeated oral feedings to rats. For single doses: Largest dose survived by all rats Smallest dose to which all rats succumbed « 0.3 g./kg. « 1.0 g./kg. Por repeated (20) doses; 0.3 g./kg. rapidly produced serious injury and death, 0 .1 g./kg. had but very slight deleterious effect and 0.03 g./kg. had none THIS REPORT IS THE PROPERTY T«fC 111». w I-J 000898C , 5068 Biochemical Research Laboratory T23.14-11-1 Page 2 *• Examination of the affected animals revealed minor effects In liver and kidneys, the major action being local Irritation In the stomach and gut. When lncorporated^lnto the diet of rats, there was no adverse effect at. concentrations which the rats would eat, 0 .03# and less. At concentrations of 0.1# and more, the rats refused to eat. The dichlorophenoxyacetic acid had but very slight effect the skin of when tested In solution upon/animals. KO There was no evidence of absorption through the skin. In making and processing 2,4-dlchlorophenoxyacetic acid there appears to be no serious hazard of systemic effects. c/i cn Atmos- q q cn pherlc contamination by dust offers the only concern and this would have to be quite severe to be excessive. Any such excessive exposure would probably be first Indicated by symptoms referable to irritation in the xipper respiratory passages or by gastrointestinal disturbances. By comparison with other common substances, it offers very little hazard of serious organic injury, even from repeated exposures. Animal results indicate little irritating action upon the skin. There is the possibility that prolonged heavy exposures to solid or to strong solutions could cause a dermatitis. This applies to both the free acid and the sodium salt. cc-c in the use of 2,4-dichlorophenoxyacetic acid, the solution concentrations are sufficiently low and the total amounts Involved sufficiently sma^l.-that there appears to be no hazard to the appli­ cator or to farm animals. 5069 0008987 ec*8 T + :iit Mi i.t .l CU IL M , 8. » T ill M F-, On i ( < u \ cn o ü o I i l Y-\ of i sonic trie recordn.0 , d ie m ovem ent o f the pin under a full load o f 1500 g . Is lim ited 0 0.04 m m. T h e bridge signal w as fed through a direct-coupled am plifier to a atlrodc ray oscillosco|K for photographic recording. T h e bridge o u tp u t w as linear hrough full-scale deflection. R estin g tension o f th e m uscle w as m aintained a t a nnslant value throughout any on e experim ent, b u t varied from on e exp erim en t to nothcr betw een 75 and 200 g. For indirect stim ulation the sciatic nerve w as crushed and tied proxim ally. T h e istal segm ent was placed in a p lastic insulating bed containing tw o silver stim u latin g cctrodcs. D irect stim ulation w as cITccled through steel needles placed a t op posite id s of the m uscle. Stim uli, w hich were supram axim al in in ten sity , were presented s square w aves generated by a trigger circuit, th e duration of which w as controlled y a variable resistance-capacitance circuit. N erve action p otentials were recorded through a Sherrington electrode from the Dial nerve isolated peripherally. Stim ulation w as effected a t the p elvic en d o f the :i'alic nerve which w as isolated centrally. F leetrom yogram s were recorded through steel needles insulated to w ith in 1 to 2 nn. of their tips and placed in th e b elly of the m uscle ap proxim ately 5 m m . a p art, lie potentials were fed through a condenser-coupled am plifier to the ca th o d e ray icilloscopc a ml recorded photographically. W hen the needles were inserted sh ortly ter section of the sciatic nerve, spontaneous random spike p oten tia ls were observed, hese spikes lessened in frequency and then disappeared in the course of 10 to 15 inutes. N o studies were carried ou t until spontaneous a ctiv ity had subsided. Intra-arterial injection w as m ade through a sm all cannula tied in to th e contratcral iliac artery and directed cen trally. T h e crural artery of th e leg under stu d y as ligated. T h e aorta was occluded during injection b y draw ing up on a loose jalurc placed ab ove the bifurcation. T he injection volu m e w as 0.15 m l. T h is eparalion preserved normal blood supply to the m uscle under stu d y b u t is under mtinuing developm ent to attain further restriction o f the in jection to th e triceps rac. T h e sm all size o f the rat’s peripheral vessels enabled th e use o f a sim p le highcqucncy d esiccatin g apparatus (T lyfrccator’, Bircher) to coagu late v essels and, i u s , to reduce bleeding. When required, denervation of th e triceps surae w as perform ed 10 d a y s before cording by asep tic section of tbe sciatic nerve. Concentrations of potassium in scrum and m uscle were determ ined in control its of the sam e w eight in whom eq u ivalen t am ou n ts of KC1 had been in jected intracritoncally (i-p ). A nalyses were m ade in .a m odified B crry-C happcll-B arnes in:rnal standard flame photom eter (9, 10) . Prior to adm inistration of curare the trachea w as cannulatcd and artificial resiration provided b y a pump. The concentrations of the agen ts used and their dosages follow : 1) Sodium pentobarbital, i f n ig /m l., tvas adm inistered i-p in doses of 40 to 50 m g /k g . b o d y eight. a) Sodium a , 4 -diclilqrphi-noiyacctate m onohydrate (R a k e r) w as dissolved in w ater in a con* miration of j o m g/m l., and the f n w as adjusted to 7.4 w ith dilute I I C I for i-p in jectio n in doses of » to a jo m g /lg . 5071 L ’ * . . j) d-Tubocurarinechloride* wasInjected ¡-pindose*of 1 .j mg/kg. Thislsequ' Happroxt* matcly to one unit of cuiare per animal and produced total paralysis of all skeletal muvtie. 4) KCI was injected i-p as • ao mg/ml. (170 mEq/l.) aqueous solution. Doses were 400 mg/kg. (5-4 mEq.). j) Quinine dihydrochloride was injected intramuscularly in aqueous solution of 45 mg/ml. and in doses of 150 mg/kg. 6) Disodium d-l-a-locophcryl phosphate* was injected i-p in an aqucouS solution of too mg/ml. in doses of 1 gAg. 7) Magnesium sulfate was injected i-p in doses of 150 mg/kg. 8) Calcium gluconate was injected i-p in doses of too mg/kgRESULTS O ur observations a re in full agreem ent w ith those of B ucher on th e b eh avior of th e ra t which h as received 2 , 4 -D ( 4 ). In b o th conscious and an csth ctiscd ra ts full developm ent of m yotonia ap p eared some 3 0 to 4 5 m inutes a fte r i-p injection and lasted for hours. T h e injection of sm all am o u n ts (2 m g.) of 2 , 4 -D in to th e vascular tree w as followed b y extrem e generalised m yotonia in tw o m inutes. E ven under deep anesthesia an d full cu rarisation, th e resistance to passive m otion of th e ex­ trem ities was appreciab ly enhanced. P rim ary cfTccts of 2 , 4 -D on muscle function arc illu strated in figures 1 an d 2 . Increase in tension developed b y an isom etric tw itch, in response to a single su p ra­ m axim al stim ulus to the nerve, was of the o rd er of 2 5 to 3 0 per c e n t; d u ra tio n of tw itch until half relaxation was g reatly prolonged (figs. 1 A a n d 2 A). T h e electrom yogram of norm al muscle stim u lated b y a single shock to the nerve is a sim ple diphasic deflection (fig. iB ). W hen m yotonia had developed a fte r in ­ jection of 2 , 4 -D th e response becam e repetitive. T h e rapidly recurring, brief p o ­ tential changes were less th a n th e initial spike, a n d occasionally th ey did n o t ap p ear u n til a fte r a sh o rt period of electrical silence, ro to 50 msec., following the initial spike. On one occasion the silence persisted for 600 msec. (fig. 3 C ). In some records the regular rh y th m icity of th e repetitive response suggested th a t a single m otor u n it was firing d irectly u n d er th e recording electrodes. T h e d u ra tio n o f re p etitio n was variable, ranging from 10 0 msec, to 6 sec. Prolongation of tw itch a n d rep etitiv e firing decreased rapidly w ith repealed stim u latio n ; a t a stim ulus ra te of 1 2 p e r m in. m yotonic features dim inished rapidly during the first five or six consecutive single volleys (fig. 1 A, B ) . . R est for 1 0 m inutes resulted in com plete re tu rn of the m yotonic response. T h e sam e changes were noted following stim u latio n by a p air of nerve volleys delivered a t sh o rt intervals. M yotonia in m an an d g oat is characterised b y exquisite sen sitiv ity of m uscle to m echanical stim uli. 2 , 4 -D produced in the r a t th is sam e explosive electrical re­ sponse to tap p in g of th e m uscle or tendon an d also to th e insertion o r m ovem ent of the recording electrodes (fig. 2 ). T h e site of developm ent of th e m yotonic response to 2 , 4 -D w as delim ited p a r­ tially by injection of sufficient d-lubocurarinc chloride to block com pletely an y muscle * Generously provided by E. R. Squibb and Sons, Mew York, N\ V'., and by Abbott Laboratories, North Chicago, 111. 4 Generously provided by IIofinian-LaRochc, Xutley, X. J. £ m j 5 l M 00 0007163 F ig . ( . I ndirect stimulation o r the triceps surae bv the sciatic nerve . A . r . N orm a gram . s - j . C o n se cu tiv e m yogram s, 110 m in . a fte r 40 mg. o f J , 4 - D , in response to stim u l ■eicd a t a rate of la / m in ., illustratin g decreased duration and tbe phenom enon of ‘w arm -u p ’ . Im e : Jo m sec. 7 . T e n sio n : ao o g . B. 1 . N o rm a l electrom yogram , a - 5 . C o n se cu tiv e records 1 A . 6 . V oltag e: a o o /0 m g. q u in in e . 4 . $ m in. afte r ad d itio n al 10 mg. q u in ine. $. 10 m in. afte r 4 . 6 . T e n sio n : 300 g. 7 . R e stin g te n sio n : 100 g. 8 . T im e : 50 m sec. C. 1 . N o rm a l clcctro m yog ram . a & 3 . C o n se cu tiv e sw eeps 40 m in . after 30 mg. a , 4 - D , illu stra tin g a n u n u su a lly prolonged period of silen ce (600 m se c.). 4 . 5 m in . afte r 40 mg. q u in ine (irre g u la rity of trace is ow ing to 1 ao cycle in terferen ce). 5 . V o lta g e : 1 >tV. 6 . T im e : 50 m sec, (referring to 1 - 3) . 7 . T im e : s o m sec, (referring to 4). F ig . 4 . D iphasic action potential from tibial nerve , i . N o rm a l, a. i j m in . afte r 30 mg. a , 4-D . 3 . 60 m in . a fte r a , 4 - D . 4 . V o lta g e: aoo jrV . 5 . T im e : 30 m sec. m inutes. T h is increase in serum con centration presum ably w as reflected rapidly in the in terstitial fluid surrounding the m uscle cells, b u t a n a ly ses o f the to ta l m uscle itself revealed no sign ifican t increase in tota l K co n ten t. W e h a v e confirm ed HZTTS l Mflfl '.‘A. m u iile in response to a sin gle stim u # ___________ — ____ II course o f response to a , 4-D pro* . ; ;nttatiori o f th e m yoton ic response an d in degree o f rep etitiv e firing ( f i g n s B ) ^ ¡Jrttr&sed se n sitiv ity to acetylcholine w as d em onstrated b y in jectin g to o ¡ig. intftu Hally ( u ) . In th e preparation described here, th is injection elicited a few sm all tered^spike p o ten tia ls in the electrom yogram . 4 A fter tre a tm en t w ith a , 4*D the; e am ount o f acetylch olin e ev ok ed a n exp losive an d prolonged burst o f electrical v»ty-4. :iV. ;* I .. . • Q uinine h as b een show n to reduce or ob literate th e m yoton ic response occurring U ancously in m an ( 13, 14) and goat (6, 8) . T h e sam e effect w as produced on itonia induced in th e rat b y a , 4- D . T h e m yogram revealed a low ering o f tension »wing adm inistration o f quinine (fig. 3B ). T here are certain characteristics of response w hich w arrant notin g. F irst, tension developed after the second dose uinine fell to 54 per cen t o f th e con trol tw itch ; and second, th e duration o f the ch w as n ot reduced. T he effect o f quinine on the responses induced b y 2, 4-D town in figure 3 C where the characteristic repetitive firing o f th e m uscle was Lerated. Alpha-tocophcryl phosphate, w hen injected i-p in relatively large d oses in to normal , produces profound generalized effects: som nolence, ataxia, m uscular flaccidity, a and, occasionally, death follow ing con vu lsion s ( 15) . W hen adm inistered per* ly to tw o p a tien ts w ith m yoton ic d ystrop h y a su ggestive decrease in spontaneous Ionia w as observed ( 16). In jection of a-tocoph cryl p hosp hate in the rat, which received 2,-q D , obliterated th e evidences of m yoton ia a s effectively as quinine. Similar suppression of repetitive response follow ed injection of magnesium and urn. D ISCU SSIO N ^Thc neurom uscular apparatus o f th e rat, treated w ith 9 ,4 -1 ), m im ics faithfully characteristics o f m yotonia occurring spontaneously in m an and g o a t. In both uhfcUnccs, m uscle exhibits exquisite se n sitiv ity and rep etitive response to several iS iitlr . . . . fcpcU tive phenom enon. A lpha-tocopheryl p hosp hate now m a y be added to the of agents inhibiting repetitive responses in both sp ontaneous and induced »iònia. T hese effects o f 2 , 4-D , producing a m yoton ic reaction so rem iniscent o f the ltapcously occurring forms, are indistinguishable, likew ise, from th e consequences càtmcnt with a variety of substances; e.g., th e ‘vcratrin c’ alkaloids derived from Urum and Schotnocaulon, aldehydes, tetraeth yl am m onium ions, phcnanlhrcne-9* oxylic acid and v ' 1 M' 1' substituents, and dihydronaphthacridine carbonic 0007165 acid ÎFF£( jcn co ïis group xVe h av e added recen tly p cntam eth*: ÎM ahy, attd pcrH api'all, o f th ese d iverse veratrinic substances, h ow ever, p rod uce u m ê -iiq iè titiv é 'm p à iü e In n erve th a t th e y produce In m uscle. I t h a s n o t b een 1 'established in Spontaneous m yoton ia w hether or n o t th is rep etitiv e p henom en on ^occurs in n erve a s w ell a s in m uscle; nevertheless, som e observations h a v e su g g ested &£*.that m y o ton ia in m an is accom panied b y functional changes n o t lim ited to m uscle fiber proper ( 19) . In deed, it is reasonable to view repetition a s a stereo ty p ed re* '> S'f'gponse of excitab le tissu e (nerve, neurom uscular ju n ction and m uscle) to m a n y un* . ■' related a ltera tiv es w hich m ay p la y râles o f differing in ten sity on a ll ex c ita b le stru c• turcs. A ten ta tiv e exploration o f this p ossib ility m ight Begin b y ex a m in in g th e circum stances in w hich repetition h as been observed. In order to lim it th e an alysis, on ly those instances of repetitive response to sin gle stim uli h a v e been considered; ob viously, this lim itation excluded o th er im p ortan t changes in ex c ita b ility m anifested b y spontaneous firing or b y m easurable ch an ges in pre-discharge conditions, such as threshold, resting p oten tial, recovery cy c le etc. ( 20, 21) . T h e appearance in excitable tissue of rep etitive, rh ythm ic responses to a single stim ulu s occurs under the m ost diverse o f circum stances, a s for exam p le: a) increased external h yd rostatic pressure ( 22); b) stim ulation b y co n sta n t rectangular currents ( 23); c) increased external concentration o f 11+ ( 20, 24) , K + ( 12) , a ceta te, lactate, citrate and oxalate ( 24) , B a ++ and guanidine ( 25) , ad en osine triphosphate ( 26), D D T and quinoline ( 27) ; d) decreased external con centration o f Ca++ ( 28); t) inhibition o f cholinesterase a c tiv ity a t the neurom uscular ju n ction b y cserine ( 1 1 ), neostigm ine ( 29) and D F P (30) ; / ) tetanus toxin affecting the neurom uscular ju n ctio n (31); g) treatm en t w ith th at heterogeneous group o f substances w hich produce veratrinic effects ( 17); and h ) spontaneous m yotonia in m an and g o a t. In the face of our fragm entary know ledge of procésscs subserving excita b ility , the incongruity o f those con d itions and agents producing rep etitiven ess m akes it ex ­ trem ely difficult to ap p ly a n y unifying concept w hich can explain a ll o f the obser­ vations noted ab ove. An exam ple o f the difficulties encountered in a sim p le approach is contained in an a ttem p t to an alyse the phenom enon o f rep etitiven ess in term s o f distortions o f cation ic vtilicit. Preservation of to ta lly normal fun ction d ep en ds on m aintenance o f con centration s of N a +, K+ and Ca++ w ithin certain lim its. C ould these m any agen ts a c t b y disturbing th e required cation ic patterns? S im ple in­ spection of the structure of th e veratrinic agents reveals their variation through a l­ m ost full scale w ith respect to chem ical a ctiv ity , steric and p hysical characteristics; some are p oten tial m etal séquestrants or précipitants, others to ta lly in a ctiv e. L ack of an y com m on characteristic is em phasized not to im p ly that a unitary' p attern o f action is n on-existent b u t to stress the necessity for fittin g m any d iv erg en t ob ser­ vations in to a n y proposed schem e. . I t seem s lik ely that these m an y different forces and agen ts m a y a ct a t different loci in the com p licated train o f even ts responsible for the sm ooth ly integrated flow of energy w hich m aintains norm al excitability. O n ly vagu e d u e s ex ist to su ggest certain possible sites o f action . F o rex a m p le Lorentc d e N 6, from an elaborate an alysis in frog nerve, su ggests SJTfSTTdl Ilflifl Sfebin erlfii m i «tudyjbf p lan ts and m icro-organism s com es su gg estiv e,ev id en ce' |-D *diitorts m echanism s in volved in th e transfer o f oxygen (32, 33) ^ C om p atibly th,J>ut n o t proving, th is suggestion is th e suppressive action o f M g * + o n veratrinic cnomena ( 24) and the sim ilar a ctio n o f o-tocopheryl p hosp hate w hich is so p o te n tly •:/! lioxidative a s w ell a s an tip roteolytic ( 15) . , , . ¿.M odels of ex cita b ility or en ergy transfer expressed In term s o f ch em ical o r p hys1 structure, en zym atic a ctiv ity , surface a ctiv ity or m em brane p o ten tia l, m a y exlin Some or m a n y of th e observations; b u t none, as y e t, h a s been offered in sa tii:tory interpretation o f all the d a ta . , Jv.The phenom enon o f the silen t period intervening b etw een th e norm al single ke potential and th e outburst o f repetitive spikes follow ing treatm en t w ith 2, 4-D the sam e a s th a t described b y E ichlcr in the frog treated w ith sm all am ou n ts of :ratrine* (34) . A sp cculative'in tcrp rctation o f this phenom enon m ig h t assum e ponential d ecay of facilitating and depressing processes in itiated sim ultaneously the stim ulus (35). If under th e observed circum stances the depressing process caycd so rapidly th at the facilitating process remained unopposed, then a b urst of tivity m ight be released. T h e d ata available do not perm it a m ore specific analysis. S U in iA K Y ' 2, 4-dichlorphcnoxyacctatc produces in the neurom uscular apparatus o f th e rat x ra tn n ic response m arked b y repetitive response to single stim u li in m uscle and rve. T h is results in increased ‘twitch* tension and prolonged ‘twitch* duration. iCrepetitive responses and their sequelae are accentuated b y K + and acetylch olin e d arc obliterated b y a ctiv ity , quinine, Mg++, Ca++ and a-tocop h eryl phosp hate, esc phenom ena arc indistinguishable from those occurring in th e sp ontaneous m yoiia o f m an and goat, and also in response to several apparently unrelated chem ical, ysical and electrical agents. Si We ate deeply indebted to Dr. S. A. Talbot for designing and supervising construction of the cIronic instruments used in these studies. REFERENCES >N, W. A., R. E. S lade and W. G. T empleton. Brit. Patents 573, 919, 1941. . nhitauAN, P. W. and A. E. H itchcock. "Cantrib. Boyce Thompson Inst, tt: j i t , 1941. ^Thompson, If. E., C. P. S wanson and A. G. N orman. Bet. Cot. 107: 476,' 1946. j B uche*, N. L. R. Proe. Soe. Exp. Biol. Jfc MeJ. 63: T04, 1946. l-T oomsen, J. A rth.f. Psythial. 6: 706, 1876. . B roun, G. L. and A. M. H arvey. Brain 61: 341, 1939. .'C lark, S. L., F. If. L uton and J. T. C utler. J . Ken. iftn l. Dit. 90: 297, 1939. | K ou , L. C. Bull. Johns ilapUns IJosp. 63: rrt, 193S. .■ Folk, B. P., K. L. Z ikrler and J. L. L ilientiial, J r. Am. J. Physiol. 433: 381, 4948. i’ L uientiial, J. L., J r., K. L. Z ierler , B. P. Folk and M. J. R ilev . In preparation. . B roun, G. L., II. II. D ale and \Y. F eldoerg. / . Physiol. 87: 394, 1936. . Walker, S. M. Am. J. Physiol. 149: 7, 1947. . Kolb, L. C., A. M. H arvey and M. R. Wiiiieiiill. Bull. Johns Hopkins llosp. 6r: *88, 4938. . Wolf, A .. Arch, i f enrol. Psythial. 36: 381, 4936. » 5074 t8 . ' Z i e u e r ,' K . J^-'a n d j . U L r u E N T iiA L , J r .' i / ^ R r a v m , O . a n d O . H . A c iie s o m . i 8 .; E y z a o u i r r e , C . a n d J. L . L n tC N T H A t, J r . 19 / D e n n y - B r o w n , D . a n d S. N e v e r . U n p u b lish ed o b s e r v a t i o n s ^ 1 V i^ ' v :V.'- 7 •' *.**■ •: Physiol. Re*. ttf: 3 8 3 ,1 9 4 8 . - .., U n p u b lish e d o b s e r v a t io n s ..'* .^ ]' ' i Brain 6 4 : 1 , >941. ; Electrical Signs o f N trvoui Activity. to . E r l a n c e r , J . a n d H . S . G a s s e r . P h ila d e lp h ia : U n lv . o f P e n n a . P re ss, 19 3 7. $ • «••••< ■ s i. K atz, B . - Electric Excitation o f Nerve. London: Oxford Unlv. Press, 1939. t i. G rondfest, II. Cold S frin t ITdrkor Sym f . Quant. Biot. 4:179, 1936. • . s j. H o d g k in , A . L . . Phytiol. 1 0 7 : 1 6 3 ,1 9 4 8 . - • 14. L o r e n t e d e N6, R . . A Study o j Nerve Physiology. Studies Rockefeller In st. 1 3 1 - 1 3 * . *5. D on, F . T . and T . P . F eno. Chin. J . Physiol. 1 5 : 4 3 3 ,1 9 4 0 . *6. B uckthal, F ., A . D eutsch and G . G . K nappeis. Acta Physiol. Stand. 8 : 1 7 1 , 1 9 4 4 . * 7 . Gordon, II. T . and J. H . W elsh. / . Cell. Comp. Physiol. 3 1 : 393, >938. *8. K opplf.r, S. W . J . Neurophysiot. 7 : 1 7 , 1944. *9. H arvey , A . M ., J. L. L oientual ,J e. and S. A . T albot. Bull. Johns H opkins Hasp. 8 9 :3 * 9 , 7 •V ; 19 4 1. Harvey , A . M ., J. L. L ilienthal, J r ., D . G rob and S. A . T albot. Bull. Johns tlo p k in t Ilosp. 8 1 : * 6 7 ,1 9 4 7 . 31 . II arvev, A. M. J . Physiol. 98: 348, 1939. 3 1. S mith, F . G . Plant Physiol. 23: 7 0 ,1 9 4 8 . 33. Worth, W . A ., J r. and A . M . M cC abe . Science 10 S : 16 , 1948. 34. E i c i i l f . r , W . Ztschr.f. Biot. 99 : 243, 1938. 33. H arvey , A . M ., J . L. L ilientical, J r ., and S. A . T aldot. Bull. Johns H opkins Ilosp. 8 9 : 3a 547i «94«. 0G 07166 9T 7T T r :l u n n lo 507; Reference 4 «¡Id ahoek, nepectlTelp. AS of the 10 eoatrola «hewed ahoek, S died, 1 m aeverely shocked, u d the r a t «bowed m o d erate o r s l i d «h ock. Three week« later w m of tbo «arTirois of both tho 4 pcrimeatel and tha control granp w en again chaDcaged without-the drag with 0.6 cc of egg white, Lr. AU «bowed TABU 1 X rrscta o r A oK in im u rion or 16 G atina ( J U G u) or A c a rru a u eru e Acta, n Divina» Door«. • a Anarmtlacric B eocs Ma. Doaose 831 333 337 •30 041 •43 •43 347 34» •33 •38 •33 •34 •34 •3« •40 •43 •44 •40 »4« •04 13 fioiaa ia 3 i i m m «• 03 M •a 36 «3 «3 03 Mom 0# aa » aa « •a os M at •b r a • • . • • • • • • • 2 1 3 2 1 1 4 2 2 1 2 4 21 dar* later 3 4 1 #9 a o O 0 1. CanrosLL, Bsaar. BaaonorasT, i n s D., aad Good, Bosaar A. ^rec. tu*, car- «M . Jfad* 1047. 04. 261. 2. Coaran, A. F„ aad Karr. S . IL J. «*». Jfad, 1041, T7. 173. 1. D naics, C. L . IliT rnrocn, C. A , aad Bwtvr. H, r . / . rUe. /aeeaf.. 1026. 6. 427. 4. B om«ras«a, T. B ra . Beat, aeo. B id . Mad* 1040, 01« N ( 10L 6. Jaota. B. T. Brae. Amir. Pcd. «He. Baa* 1047, 0 1 «. R im . L A , Boiaraar, P. J., and trin a , W. W. Betaaee, 1040, 100, 332. 7. Bolliva*. C L P ia a n . T. V , aad Riaaaar, A W. P ra . B«e. «**. B iel Med., 1343, 07. 303. A Bwirr. H. P. J. ero. Bad.. 1322, 33. 738. cn o 00 05 ro Effects of 2,4-Dichloropbenoxyocedc Add on Chicks1 • 0 , , IfELVoi K. Bjoon and E x n r T. N o o n s» 4 4 1 Department « / Botany, Dnfearrify « / Wyoming # 0 0 41 •• To teat tho «ff«et of aeatylaalleplla ad d oa histaariaa aek, 11 rabbits weighing 1 -1 1 kg war* g in n 6 graiaa Of neetylaalieylle add, orally, a t StSO PAL oa ICareb 11, 1047. Thla doaago waa rapante* a t 0:00 AAL oa March 12 and again a t 2:30 PAL the aaam dap. One boor later, each rabbit waa Injected intravenously with 1.73 mg of hkUmiae phosphate. S e ra of tha 11 died (# 4 abode), S «bowed «arerò (# 3 ) shock, aad 3 «bowed aMdorato (# 2 ) ahoek. Tbo animala whieh wore preaedieatad with aeetplaaliepUe ad d «bowed atrikiag proteetioa agalaat anaphylactle ahoek. I t aaap bo praw ned, oa tho baaU of tbo M en to n rerlewed sbovo, th at thla waa dao to a do* errai« la the aatlbodlea a t tha time o f the ekeUengiag doae. Thero la aethiag to indicate anp protaetlro effect againat hlataalne ahoek, fo r though anaeeompanied bp aimoltanaoaa control«, the aaorbidltp la tho «boro aoriao waa d o lla r to th at reported la a predoni coauanaieatJoa (1). I t la iatam tln g to noto that the offoeta of tho drag were temporarp and th a t tho treated animala ahowed eoadderabla bppeneniitiritp 3 weoka later. Tho eontra d between tho reanlta with thla drag aad thoae reported earlier (p) fo r Beaadrpl ia completo. Benadryl gare good protection, In dm tlar «sperimenta, ngalnat hlataalne ahoek bot waa withont affect oa an­ aphylactic ahoek. Tho p m e a t drag, aeatplaalieplie add, however, effect!re Ip protect« againat anaphylactic ahoek Lat not ngalnat biatamine. I t la, with tha other aalleylCCIEN CE, O ctober 29, 194«, VoL 10« a tea, ap p a ra tip a trae antlanaphplaetle drag in th at it Interfere« with tha antigen-antibody reaction« to prerent or deereaae tha ontoward reanlta of tho challenging doae of antigen. With the inereadng n o of 2,4-D aa a harbieide it ia important to inquire iato tho poaribh toxio offoets of tho chemical oa animal«. Several larcatigatora bara atndiad tha effect« of 2,4'D oa mammal«. Tha lethal doea for mice, when injected aubentnneooolp or intraraocaly, haa been determined bp Bnehiar (1) to ba t86 mg/kg of body weight, MitebeU and K arth (B) reported that thay fad 200 mg of 2,4-D daily, to n a i l experimental aaimala with ao ill effects. TABU 1 C m n or AUAnoLAMina or 3,4-D on W arn Bocx C aicso DoMce (m s of acld/knof body welcht) 0.00 (control) J§ 3.80 28.00 280.00 < p z 9 * 1 \ > J lacera* la weight at end of fonr weoka (%> < N » m3 N 2 433 444 4M 427 373 Z la tha p ra e a t aspefimant Whlta Bock ehleka won naod. Tha dkanolamiaa of 2,4-D waa admialaterad orally through a pipette. In tho drat «apartment, data for whieh ore giro« ia Table 1, one part of tbo alkanolomlwo waa dilated with 10 parte of wnter. The dosages recorded are in terms of tho add equivalent. Five ehleka (each weighing approximately 60 gm a t tha beginning) w en used la each group. Tho chirk« w an weighed and then won g ir a tho appropriate doaa (Table I) three time« a week oa alternate dap* for n period of fonr weeks, making a total of 12 d o ra Aa the ehleka gained in weight, the o z z 0 >Coattibottoa Ma. 210 from the D eport»»« of B ettor and tha Aock? Manatala Heibarlas^.Ualvanltr of Wromlns. •XI 5076 0002798 f • » 1. B e ca a a . R . L . 1 Pree. gee. am . S M . UH.. 1040. S3, '20*. - 2. U ircaau , 1. W , aad Means, P. C. get. S u , » 4 4 ,100. too. A New Histological Procedure for Whole . Tissue Cultures Grown in Plasma A m u i i u Conan aad Coautt Watkouth Chester B m ttg Jtcsrereh /astitate, Jt syol Cancer Jgaspital, f»0tam flood, Aendoa, S .JFJ Standard taxtbook proeadaraa ( f , J ) reeeauaeaded for fixing and etaiaiag whole tissue outturn la plasma eoagula hare prorea uaaatlefmetory for the following reasons i (1) The dense fixed eoagulum presents a nearly impermeable barrier to the stale, thus aeeemitatiag pro­ longed staining periods) (2) the aoagulum itself stains diffusely and haarily, presenting adequate contrast be* twees the cells end the medium aad aeeeaeitating careful aad tlme roneualng deeolorising aad deratoplag pro* eedores) ({} the reeuttlag^preparation, when mounted, is thick, hoe o tendency to form sir bubbles, does not •T b s anthem i n sratefel to Hebert W. U adsestru tb, who msde tbs, eism lsstto es. 480 dry rsadily, end le generally Uadoquato fo r mietooeopU etndy. Tompkias, Cnnningbam, aad K lrk (d) mesutly attem pted te Imprese ternato by woohiag cattareo in a . salt salatina fo r 4 hra a t 7* C te remore eeluble p iatela before fixatioa, bat stlU had te reeert te prolongad etnialag (1 k r ia Delafleld’s kem ateaylia) followed by aer* ria l baure of waahlng. Earle (2) haa dariaed «empii* ratrd fixing, sU iaiag, nad meuaUag piocodwcs, too elab­ orate ta w arrant eraloatloa beta. t DOW 508325 •mount administered « u adjusted is ardor U m h U h the original dosage. A t tho o d e f four « w b they «am •g ala weighed. T te table «hows the perecatago lie m m is weight fer tho looM N tk period. The differences between the eeatrol group a id theee girea docagee of 0.28, S M , a id 28.00 m g/kg were le t d p ld e u t a t the 5% Im L The difference betweem the eeatrol groep a id the greap fire s • doeage ef 280 m g/kg waa barely significant a t the t% lereL Next, experiments were started to determine the lethal does of the elhaaolamiae e f 2,4-D whea dilated 1:0 with water. Each ehiek of a group of 8 (each areragiag 160 gm) wae girea one dees of 280 mg/kg of body w eight These ehleha «w eired. Each ehiek e f aaother greap e f S ehieha (arerage weight, 111 gm) waa girea a dees e f 768 m g/kg of body w eight All of these ehieha died. Postmortem examinetioa rereeled a fatty degeaetatioa with a pale se ttlin g ef the llrer, spleen, kidneys, aad h eart Hetnorrhagie gaatreeateritia waa abo erideat* Hence, for email ehieha the lethal does of dilated 2,4-D is eomewbere bo* tweca 280 and 768 mg/kg. Tho fact that a single does of 763 mg killed, bat not a total dees ef 2,260 mg administered orer a four-week period (280 mg/kg ehieha of Table 1), la diestee that the alkanolamiae of 2,4-D U not a eamolatire poison. The question might be raiaed as tc tho poeeibility of ehieha being hilled by feeding ea plaata which had been sprayed with 2,4-D. A t a spraying rate of 1 lb of 2,4-D/ aero, a ehiekea weighing 1 kg would hare to consume oil of the 2,4-D applied ea 72 sq f t within a day or two to obtain a lethal dees. P ia. 1. ntass appesisse* af staiard tteeae eeltam e: A, plsaam aadrltd : B. plasma drisd (a 1 2 ). VTe here found that most of the dlaadraatages e f the plasma eoagulum con bo eliminated by drying tho whole tissue culture preparation after fixation. The prosed ore is as follows: The culture in its eoagulum on a eqror slip ia fixed orernight in 3-4% formol containing 02% acetic aeld or la Carney's fluid for 1 h r. la general, aeetioaleohol fixatioaa giro a m art granular aad more opeqae dried specimen than aeetie-fonaol fixatioaa. The sorer slip is then washed thoroughly ia raaalag w ater (after aeetie-fonaol) or la descending strengths of alcohol, aad water (after Carney’s). A fter a final rinse ia dlitHlod water the carer slips are laid flat on a glass eurfaoa protected from dost (P etri dishes or etaiaiag dishes eaa be used) and allowed to dry slowly aad thoroughly in air. I f a preparation of a tube or a flask culture ia plasma is desired, the portion should be fixed aad washed in mtm, pried loess sad placed, like a paraffin sectton, ia a dish ef water, where it eaa ba manipulated with a dissecting needle sate a elide or eorer slip prerleueiy coated with a thin film ef M ayer's albumin, aad dried as a bore. Cultures grown ia a fluid medium should aot be dried. The dried cultures may ba stained w ith say one of the hematoxylins for from 8 tie IS a la , depending ea the strength ef tee stain. W elgert *s iron-hematoxylin has been found eery satisfactory. The staining ia simple to eoatrol after a prelim inary-trial, mad prolonged wash*, lag to derelep the color is aot necessary. Ones stained, the cultures should net be allowed to dry again. Poliowlag 'this, tee eqror allpa ere passed through the aleohola in tee usual manner, cleared iu-xyleae, end mounted. I f a I r ishman or Olomea stain ia used, acetone and acetone* xyleae dehydrations should be employed Instead ef the alcohols. The dehydration end electing procedures take considerably leas time with dried cultures, sines each SC IE N C E , O ctober 2». 1948, VoL 108* ^ q ^ rj 00027S9 5078 y 3 ^ Biochemical Research Laboratory TH E DOW CHEMICAL COMPANY «* R E S U LT 3 OP S K Z R IR R IT A T IQ R T E S T S OR VA RIO U S SA M PLES OP 2 ,* - D TA X ER A T V A RIO U S STA G ES IR T H E M ARUFACTUR2RO PRO CESS fb . T23.1U11-V CH#. 1 2 1 9 -0 2 ^ 5/53/*9 Fin’d 10/12/^0 W ork By V . X « RO V O Ckock B r it t o n 's D iv is io n A t t i J . V . B r it t o n R« C . D o sso r C . A . K lg h h lU H . R . H o y le /d - /¿ -4TS / E s e c u t iv o R o s e s re h C o a a it t e e T H i i ; ■ i-v.'hT Ü THE Pr DOW CMEr:CAL COMP — Y Or «.«•¿yk Ia p u r e 2 ,A - D p u r if ie d is e o n s id t r s b ly s e r e ir r it a t in g th a n th e p ro d u c t. cn 2 vt- D ie h lo ro p h e n o x y a e e t la s e id A e e tlo a e id t <0 ** >0 CD m CD o cn ir\ o 2 >t - d lc h lo ro p h cn o x y - J A fe w e a s e s o f d a m a t lt ls h a w s b e e n o b s e rv e d a ro u n d p a r t ic u la r o p e r a t io n s in th e a a n o fa c tu r e o f 2 #A » D . S a a p le s f r o n th e v a r io u s s ta g e s o f a s n u fa e t u r e w e re s u b s it t e d ir r it a t io n te s ts to f o r s k in d e t e r s in e w h e th e r o r n o t t h e r e w a s a s ig n if ic a n t d if f e r e n c e b e tw e e n th e v a r io u s p r e p a r a t io n s . M A TER IA L R a sw s 2 , A - D le h lo ro p h e n o x y a c e tle P o m u la t Cl < fi * Cl S tru e t u r a li a c id H -O-C-C -0-H H E a p ir ic a lt S a a p le s o b ta in e d b y i CgHgO ^Cl^ H . R . H o y le 0007478 f 5079 Biochemical Research Department T23.14-11-3 Page 2 Sample description: 3.R.D. K no. 2372-12 13 drier feed from plant no. 2. 267-Building containing 0.24£ of 2,4-dichlorophenol. K2372-13 is drier feed fro m plant no.l, 267-Building containing 1.2ljfc of 2.4- dichlorophenol. K2372-14 is unwashed cake fro m the filter box, 267 -Building containing 0.96# of 2.4- dichlorophenol. X2372-16 is a n especially purified sample of 2.4- D obtained from 267 -Bulldlng, low in 2.4- dichlorophenol. EXPERIMENTAL RESULTS W h e n the purified sample of 2,4-D, K2732-16, was repeatedly 00jy . 549306 K2372-15 is cake from the BIrd f ilt e r , 489Bullding containing 0.10£ of 2, 4-dichlorophenol. bandaged onto the shaven abdomen of a rabbit in a dry state, it produced no detectable irritation; w hen wet with water it produced only a very slight, simple irritation. <* When the 2,4-D cake fro m the B i r d filter, X2372-15, was repeatedly bandaged to the shaven abdomen of a rabbit, it failed to produce any appreciable evidence of irritation except a rather bluish coloration of the dermal tissue; w h e n wet it produced a slight amount of superficial denaturatlon, again with a rather marked purplish coloration. This lesion healed promptly after exposures were terminated. Whe n the dry 2,4-D unwashed cake from the filter box in the 267 - B u i l d l n g , ,£2372-14, was repeatedly bandaged to the shaven abdomen of a rabbit, it promptly produced an appreciable amount of irritation, characterized by hyperemia and exfoliation; whe n dampened with water this sample of material produced a moderate to severe 0007479 5080 iiiac hemica! T 2 3 . 14-11-3 Page 3 response. -T (2,4 , b - t rlohlotop h e n o x y a c e t l c acid) for use in the toxicological studies on aogs. The sample of 2 , 4 - D b e i n g sent la r egular p r o d uction 1natei loi a s s a y i n g 9 8 .5 ^ a n d h a v i n g a f r e e z i n g point of 132.ti*C. The sample of 2 , 4 , 5 - T being sent is a lso regular procuctl:; m a t e r i a l a s s a y i n g 100)1 a n d h a v i n g a f r e e z i n g point of 148.8Vj. B o t h o f these m a t e r i a l s a r e being s u p plied to you in the aci d form. B o t h a r e f a irly s oluble In olive oil and both cui oni Lj be put In a q u e o u s s o l u t i o n w i t h caustic. I bolieve you will fin-! that a a l i g h t e x c e s s of ca u s t i c la r e q u i r e d to put theae material;-, s o l u t i o n a n d that o n c e In solution, a large part of the e x cess al\nll c a n be neu t r a l i s e d . Hence, f o r a 5 .0^ a q u e o u s ».o’.utlon of the b o -.u .: salt of 2,4-D, the p H a f t e r s o l u t i o n is effected, can be adjusted d o w n to 7.2 w i t h HC1; w i t h 2 , 4 , 5-T the final pH of a 1.0£ aqueous s o l u t i o n can probably be i n the n e i g h o o r h o o d of 10 . W h e n 2 , 4 - D or 2,4,5-1' is used ae a n herbicide, the c o n ­ c e n t r a t i o n usually ranges f r o m 0 . 1 ^ to 0 .2,*. The a c t i v e luiteri-.l r.iay be a p p l i e d In a n y one of a numb e r of forms d e p e n d i n g upon tne particular r e q u i r e a e n t s of the job. fhe most c o mmon forms a. c- 1 . s o d i u m salt, a l a a n o l amine salts, and esters. _ ......... 09 * 1 © Almost all the toxico l o g i c a l work so far reportcu ^ w i t h 2 , 4 - D a d m i n i s t e r e d e i t h e r as the free acid In oI.L or r.' ■ s o d i u m salt In a q u e o u s s o l u t i o n at a pil of ( . 2 . *e navt to... r. . a fair a m o u n t of toxicol o g i c a l work on the other f«:rr.iO of L - a l s o on 2 ,-*.S>-f in its various fornn. V.e nave outervc=; ii.di. . 5083 of the ma t e r Lai :u*hei> little d i f f erence to u.u- .* ..),<•. ue«.. ;.ni .; that, acutely,, u.ere is wot u.ucn d i f f erence j .¿1 0 , . .... Dr. V. A. Drill Pig« 2 December 13, 1949 The toxicity picture as jre Jiaee At Am *i»sn An Atom following tabulations! Toxicity of 2.4*0 when administered in Single Doses. Route Rat Rat. Oral Oral Mouse Mouse Guinea Pig Chloks Aoorox. LD~0 (a.Ax.) Source Oral Sub-Cut. Oral 0.3 - 0.5 0.280 Lit. 1.000 Na Salt Dov - Incomplete Na Salt ACld Lit. Na Salt Dow - Just started Oral 0.666 Lit. Na Salt Dow-Incomplete Na Salt Acid Isopropyl esters Lit. Na Salt Dow - Incomplete Acid Lit. Na Salt 0.70 ^ o n n n Animal 0.5 - 0.6 0.5 - 0.7 0.375 0.5 - 0.7 0.3 - 0.7 0.38 - 0.76 Toxicity of 2.4-D when administered in Reseated Doses. Animal Route Source Notes Dogs Sub Cut or I.V. Lit. Na Salt 0.200 g./kg«/d*y-death in 2 da, 0.100 g./kg./day dsath in 2-4 days 0.030 g./kg./day dsath in 3 days 0.023 S « A 6 */d*y marked affsets Guinea Pigs Orally (/) Lit. Orally in diet Lit. Dow I.P. Lit. Na Salt 0 .1 0 0 g ./ k g '/ d * y 10 times in 12 uaj was tolerated O.lji tolerated 0 .1 £ -caused very slight effects 0. 03^ clear No marked histop&tftolSgical * Rats Mies C G O 'S £ 2 3 findings ho ne o p l a o t i c ¿rowtna R e p r o d u c t i o n G.K. 5084 Page 3 Dr. V. A. Drill December 15, 19*9 Toxicity of 2*4-D jibán ad*inlstered in -Repeated Dotas. - Cont *d Route Chicks Orally Lit. 0.028 «.A«. 3 tlnaa/vaak for 4 weeks - 0.1 . 0.280 g./kg. 3 tinaaAaak for 4 waakt - growth daprattlon Orally In Diet Dow 0.1* for 1 waak - O.K. 0.3* for 1 waak - O.A. azcapt for slightly dapraated growth rata. o Motes Source Anlaal Toxicity of 2,4,5-T Whan Adainlttarad in Singla Dotat Orally (This Information It from our data and It not eoaplata - We art finishing It.) Anlnal Rat Wo tat LD30 for the acid H a t between 0.5-1.0 g./k£ LD50 for the isopropyl attar lias between 0.3 - 1*0 g.Aff* Mous« Oulnaa pig Chick LD30 for the aeld lias between 0.3 -0.7 g . A s LD50 for the aeld lias between 0.3 -1.0 g. A s Por both the aeld and Wa salt Work Just begun Neither 2*4-D nor 2»4,5-T are aore than vary wild tkin Irritants «van in eonoantratad fora. In dllutad fora ready for uaa, they present no health hasard. Studies eonduetad upon guinea pigs by Hill and Carlisle on tha Inhalation of 2,4-dlchlorophenoxy aoatlo acid dust, either wat or dry, but of unknown particle site. Indicate that tha dust It not likely to causa aystaale Intoxication. This hat bean born out by our own ex­ perience with nan handling tha dust. Tha dust, however, it capable of causing Irritation of the note if the concentration in tha breathing zone becoast sufficiently high. ..... Tha following references nay be helpful if you wish to consult the literature for a aore detailed description of procedures, etc.: CCO-iT-iO Dr. V. A. Drill Page A December 13» 1949 1. Toxicity of 2#4-Dlchlorophenoxy Acetic Acid for Experimental. Animals. Edwin AT. Hill and Harold Carlisle (Camp Detrick, Frederick, MD.) J. Ind. Hyg. Toxicol. 29» 85-95 (1947) C.A. All 3217a (**y 20, 1947) 3. 2,4-D Toxicity. I. Toxicity Towards Certain Species of Fish. Jos. W. E. Harrlsson and Edward W. Reas (La wall and Harrlsson, Research Consultants, Philadelphia, Pa.) Am. J. Pharm. 118, 422-5 (1946) C.A. 41 (2529) (1947) 4. Effects of 2,4-Dlchlorophenoxy Acetic Acid on Experimental Animals. Nancy L. R. Bucher (Harvard University) Proc. Soc. Exptl. Biol. Med. 63» 204-5(1946) C.A. 41* 808 (1947) Data on the acute oral toxicity of both 2,4-D and 2,4,5-T for dogs Is lacking. Therefore, It is suggested that the acute oral studies be started at your earlleat opportunity so that Information will be available for choosing appropriate dosage levels for the 90 day feeding tests. In the single dose experiments we suggest that a notation be made regarding any symptoms exhibited at the various dosage levels, body weights for each day during 3 or 4 days Immediately following administration and on alternate days during the rest of the 2-week observation period. We do not feel that It Is neoessary to do any elaborate tissue studies on acutely dosed animals. We would, however, like to see animals that die and those that survive autopsled and the gross condition of the Q.l. tract, liver, and kidneys noted, A few sections at critical dosage levels might also be desirable - at your discretion. CGG4c4i \. 5086 i n o n o 2. Tolaranca of Farm Animals to Faad Containing 2,4-Dlchloro phenoxy aeatlc aold. J. V. Mltchall, R. E. Hodgson and C. F. OastJens (U.S. Dapt. of Agr., Washington, D. C.) Animal Scl. 3» 228-32 (1946) C.A. 40 (17); 5198. (Sapt. 10, 1946) Pag« t> December la, 194y AArSn Dr. V. A. Drill In the 90 day faading tests, we suggest that the material be administered with a part of the diet. Dosage levels cannot be decIdea upon until acute data la available. We would suggest that general ob­ servations, growth records, and periodic hematological examinations (initially, after 30 days, and after 90 days on the experiment) be con ducted during the experiment. At autopsy, gross and hlstopathologlcal studies on the liver, kidney, stomach, intestine, spleen, lung, heart, adrenal, testis or ovary, and possibly brain and vascular system are of Importance. We believe that negdttW^rfWIngs at upper dosage levels make similar studies at lower dosage levels unnecessary. The requisition for this work has been approved and I will see that an appropriate check is deposited as before. If there aze any details which are not clear or if any problems arise, please contact us at once. I regret that I did not have more time last week to spend with you. Nevertheless, I was very glad to get home early and avoid the snowstorm that settled on this area about 5 t00 p.m. that evening. Please give ay regards to Dr. Hays. Sincerely yours, THE DOW CHSHICAL COMPANY / c <> V. K. Rowe Biochemical Research Department VKR/uig '42 5087 ACUTE ORAL TOXICITY OP 2,4-DICHLORO- PHENOXYACETIC ACID TO RATS, MICE, GUINEA PIGS AND CHICKS J - £ E 8 Q S M 0 9 Reference 6 V. K. Rowe D. D. McCollleter and H. C. Spencer Biochemical Research Laboratory The Dow Chemical Company Midland, Michigan 5089 0002800 -1The 2,4-dichlorophenoxyacetic acid was administered by oral intubation to rats, mice, guinea pigs and chicks as a solution in olive For comparison, the sodium salt of 2,4-dlchlorophenoxyacetic acid was fed in aqueous solution to rats and guinea pigs. The. guinea pigs and the white rats were mature, young adult animals selected from the stock colonies of this laboratory. Young 0oV/508335 oil emulsified with 5-lOjS gum arable solution. adult white mice were obtained from the Carworth Farms, ROckland, New York. The chicks (New Hampshire Red) were purchased from a commercial hatchery and used when about three weeks of age. Mixed sex groups of each species were used in these tests. Table I summarizes the concentrations and volumes of olive oil solutions of 2,4-dichlorophenoxyacetic acid administered to each species. TABLE I SINGLE ORAL ADMINISTRATION OF 2,4-DICHLOROPHENOXYACETIC ACID Species Per cent Material in Olive Oil Rats Guinea Figs Mice Chicks 5, 10 10 5, 10 3 Volume Given by Stomach Tube Minimum (ce.) Maximum (cc.) 0.4 0.9 0.1 0.3 4.0 2.9 0.22 5.2 Mortalities resulting from the various single oral dosages of 2,4-dichlorophenoxyacetic acid and its sodium salt are given in 5090 0Ö02801 Tables II and III. All surviving*animals were observed until it was certain that they had fully recovered (usually about two weeks). Symptoms of poisoning, particularly with the higher doses, were ataxia TABLE II MORTALITIES RESULTING FROM ADMINISTRATION OF SINGLE ORAL DOSES OF 2,4-DICHLOROPHENOXYACETIC ACID Dosage (gm./ k g m .) Rats No. Fed No. Died 10 10 10 10 10 10 3 0 0 0.1 0.2 0.3 0.4 0.5 0.6 0.7 1.0 Guinea Pigs No. No. Died Fed 3 6 7 10 3 No. Fed Mice. No. Died 4 6 5 6 6 0 1 4 6 6 6 6 6 0 6 6 Chicks No. No. Fed Died 6 0 6 1 6 6 6 6 4 6 eoW 508336 and myotonia. TABLE III MORTALITIES RESULTING FROM ADMINISTRATION OF SINGLE ORAL DOSES OF 2 , 4 -DICHLOROPHENOXYACETIC ACID, SODIUM SALT Dosage (gm. A s m . ) No. Fed No. Died .0.3 0.5 0.7 1.0 10 10 10 14 0 2 4 14 Rats . Guinea Pigs No. Fed No. Died 6 6 6 0 4 6 The acute oral L D ^ q values with their 19/20 confidence limits v:ere determined for each according to the method described by Litchfield and Wilc o x o n 1. The results of the statistical treatment of the dosage- response data are presented in Table IV. 5091 1. Litchfield, J. T., Jr. and Wilcoxon, F.: J. Pharm. and s 1 -3- TABLE IV THE ACUTE ORAL LD50 VALUES FOR 2,4-DICHLOROPHENOXYACETIC ACID WHEN ADMINISTERED TO RATS, MICE, GUINEA PIGS AND CHICKS LD50 (19/20 Confidence Limits) Slope Species F orm Rats Mice Guinea Pigs Chicks Oll-acld Oil-acid Oil-acid Oll-acld 0.375 (0.302-0.465] O .368 :0 .3 12 -0.434 0.469 0 .397 -0.553 0.541 [0 .358 -0 .8 17 ] 1.6 2 Rats Guinea pigs Aq.-Na salt Aq.-Na salt 0.805 [0 .610 - 1 .063 ] O. 55 I [0 .4 17 -0 .727 ] 1.77 1.55 gm. A g m . Functi *The fold change In dosage required to produce one unit standard deviation In response along the line. DOW508337 1.23 1.17 I. 8 I The Litchfield and Wllcoxon test for parallelism of two lines revealed that the dosage-response Mine for the rats (oil-acid) deviated significantly ( 19/20 probability) from those for the mice and the guinea pigs receiving the 2,4-dichlorophenoxyacetic acid In olive oil. The line for the chicks also deviated significantly in parallelism from those for the guinea pigs and mice. The tests for parallelism of two lines and estimate of relative potency Indicated no statistically significant differences in comparison of rats versus chicks and mice versus guinea pigs. The dosage-response curves for guinea pigs (oll-acld) and guinea pigs (aqueous-sodium salt) did not deviate significantly In parallelism or relative potency. A comparison of the rats (oll-acld) versus rats (aq.-Na salt) revealed no significant difference In parallelism of the two lines,but the test for relative potency Indi­ cated that the free acid was significantly more toxic to rats than was the sodium salt. 5092 0Ö028G3 to '■ G ( 5 0 9 3 VO SEMICHRONIC ORAL TOXICITY OP 2,4-DICHLOROPHENOXYACETIC ACID TO RATS Oral Administration - Repeated Doses by Stomach Tube. II. Oral Administration in the Diet BOW 508338 I. V. K. Rowe D. D. McCollister and H. C. Spencer Biochemical Research Laboratory The Dow Chemical Company .. Midland, Michigan 5094 0002804 i / - DOW 508339 h 1- I. ORAL ADMINISTRATION - REPEATED DOSES BY STOMACH TUBE Experimental Procedure Thirty-one female rats, 60 to 80 days old, were divided according to body weights into groups of five each (six in the control group) and \ maintained on the stock rat diet. Before the repeated oral administration of 2,4-dichlorophenoxyacetic acid was started, weight records were kept on the animals for a period of 21 days to make certain that the groups were well matched in respect to growth. The rats in each of the groups were given by means of a stomach tube repeated oral doses of 2,4-dichlorophenoxyacetic acid dissolved in olive oil and emulsified in 2 to 3 cc. tion. of a 5-10 per cent gum arable solu­ The dosages fed and the olive oil solutions used are given in Table I. TABLE I DOSAGES ADMINISTERED AND OLIVE OIL SOLUTIONS OP 2,4-DICHLOROPHENOXYACETIC ACID USED Dose g.Ag. 0.0 O.OOJ 0.01 0.0'3 0.1 0.3 Per cent 2,4Dlchlorophenoxyacetic acid in Olive Oil Approximate Vol. of Olive Oil Solutlon Administered per Rat per Dose 0.00 1.5 1.0 1.0 1.0 1.0 1.0 0.06 - 0.20 0.60 2.00 6.00 5095 0002805 The animals were weighed at intervals of two and three days each group. Hematological examinations were made on the control rats and on those that had received 20 oral doses of 0.0? and 0.01 g./kg. 2,4-dichlorophenoxyacetic acid. At the end of the experiment, all of the surviving rats were killed by decapitation, and oxalated blood wa collected for urea-N determinations.^1^ DOW 508340, during the experiment, and a record was kept of the average growth of Each rat was examined for gross pathological changes, organ (heart, liver, kidneys, spleen) weights were obtained, and tissues were saved for microscopic examination. Hema- toxlln and eosin-stained sections of the following organs were prepared: lung, heart, liver, kidney, spleen, adrenal, pancreas, stomach and intestines. EXPERIMENTAL RESULTS The rats that received 20 oral doses In 28 days of 0.00?, 0.01, and 0.0? g./kg. 2,4-dichlorophenoxyacetic acid showed no 111 effects so far as could be judged from gross appearance and behavior, survival, growth, hematology (Table II), blood urea-N values (Table III), organ weights (Table III), gross appearance of organs, and hlstopathology. (1) Barker, S. B. and Urine. The Direct Colorimetric Determination of Urea in Blood J. Biol. Chem. 152: 45?-46?, 1944. n i .TABLE II AVERAGE HEMATOLOGICAL VALUES OBTAINED ON FEMALE RATS THAT HAD RECEIVED 20 ORAL DOSES IN 27 DAYS OF 2,4-DICHLOROPHENOXYACETIC AC I D Dose » g.Ag. Controls 0.0? 0.01 6 11.4 13-9 18.8 5 10.5 13.3 1 8 .> 5 10.0 12.9 17.4 10 90 15 85 9 91 Number of rats in group Erythrocytes (million per cu.mm.) Hemoglobin (g./100 cc.) Leucocytes (thousand per cu.mm.) Differential Count: r.Neutrophils (per cent) Lymphocytes (per cent) TABLE III AVERAGE BO D Y WEIGHTS* ORGAN WEIGHTS A N D BL O O D UREA-N VALUES OF FEMALE RATS THAT RECEIVED REPEATED ORAL DOSES OF 2,4-DICHLOROPHENOXYACETIC AC I D g-Ag- No. of Doses No. of Days 0.00 0.003 0 .0 1 0.03 0 20 20 20 0.1 0.1 0.3 15 15 2 28 28 28 28 21 Dose No. of Rats Surviving 21 2 6 5 5 5 3 2 2 B o d y Wt. (&) Final* Initial 195 191 191 187 190 19* 195 203 197 195 192 126 202 186 g.) Organ Weights (g./100 Spleen Liver Kidney Heart 0.42 0.40 0.42 0.41 0,51 0.43 0.48 3.40 3-37 3.38 3.25 4.68 4.08 3.0 8 0.75 0.75 0.79 0 .8 1 0.94 O .87 O .82 0.19 0.23 24.9 24.8 32.8 25.9 0 .1 6 2 6 .6 0.23 0.17 17.2 -- 0.23 0 .22 •Fasted overnight. 0Û02S07 mSOSMOQ cn o CO Blood Urea-N mg. % -4- Of the five rata that received repeated oral doaea of 0.1 g./kg. 2.4- dichlorophenoxyacetic acid, three loat weight rapidly after the flrat On the other hand, the other two DOW 508342 f ew doaea and 8howed a marked diarrhea. rata receiving 0.1 g./kg. maintained their initial body welghta and 8howed no evident 111 effecta so far as general appearance and behavior were con concerned. All of these rata were killed and autopsled after they had received fifteen doses In a period of twenty-one days. The outstanding finding In these animals was varying degrees of gastrointestinal irritation; there was also slight cloudy swelling of the liver with an increase in weight of this organ (Table I I I ) . Blood urea-N values obtained on these animals compared favorably with those for the controls (Table I I I ) . Three of the five rats fed 0.3 acid died after the second dose. g*As* 2,4-dichlorophenoxyacetic Consequently, the two surviving animals were killed and examined; severe gastrointestinal Irritation was observed in both of these rats. SUMMARY AND CONCLUSIONS Female rats that received 20 oral doses In 28 days of 0.003, 0.01, and 0.03 g./ks* 2,4-dichlorophenoxyacetic acid showed no adverse effects as Judged by gross appearance and behavior, mortality, growth, hematological values, blood urea-N determinations, organ weights, and gross and micro­ scopic examination of the tissues. On the other hand, rats receiving repeated oral doses of 0.1 g . A s * 2.4- dichlorophenoxyacetic acid showed varying degrees of gastrointestinal irritation with marked weight loss in three out of five animals. doses Higher (0.3 g.Ag*) w«re not tolerated upon repeated administration. 5098 0002808 -5- II. ORAL ADMINISTRATION IN THE DIET Ol : O £ Experimental Procedure ^ 00 The modified Sherman diet, which has been used s u c c e s s f u l l y ^ for several years In this laboratory as the stock ration for rats, served as the control and basic diet In this experiment. The follotving Ingredients were used In the preparation of this stock diet: Percent by weight Whole wheat, freshly ground 55 Dried whole milk 25 Dried extracted liver 12 Dried yeast 5 Iodized table salt 2 Calcium carbonate 1 The experimental diets were prepared by thoroughly mixing finely ground 2,4-dichlorophenoxyacetic acid with the stock diet on a "percent by weight" basis. No diet preparations over a month old were used. Female white rats from the stock colony of this laboratory were used In this Investigation. These animals were the descendants of rats obtained In 1938 from the Wlstar Institute. caged together In wire bottom cages. Five rats were The animals were fed from alurai- V». num hoppers which were weighed and refilled three times a week. Each rat had access to food and water at all times. 5099 nftAnoi'a -6- The rats were maintained on the stock diet fro m the time of v/eaning until they were 2 to 5 months of age, when they were divided according to body weights into well matched groups and started on the experimental diets. Groups of 5 female rats each were placed on diets 2,4-dichlorophenoxyacetic acid. All rats were weighed twice a week throughout the course of the experiment. DOW 508344 containing 0.00 (control), 0.01, 0.03, 0.10, 0.30 and 1.00 percent Records were kept of body weight, general appearance, and estimated average daily food consumption of each animal. During the course of the experiment, rats that were obviously quite sick were killed and examined to determine the cause of i llness. At the end of the experimental periods, all surviving rats were fasted overnight, weighed, killed by decapitation and examined. Certain organs (liver, kidneys, heart and spleen) of each rat were weighed and.the tissues of all animals of each group were saved for histopathological studies. Hematoxylin and eosin stained sections of the following organs were prepared: lung, heart, liver, kidney, spleen, adrenal and pancreas. In addition, stained sections were prepared from the stomachs of the rats that received the diets containing 1.00 and 0.30 percent of the test material. The concentration of urea-nitrogen of the blood was determined by the diacetyl monoxime method^ on the control rats and on those that had been maintained for 12 days on diets containing 1.00 and 0.30 per­ cent 2,4-dichlorophenoxyacetic acid and 113 days on diets containing 0. 10, 0.03 and 0.01 percent of the test material. 1. Barker, S. B., J. Biol. Chem. 1^2: ^53-463, 19^4. 5100 0 0 0 2 S 10 -7Exuerimental Results OOV/ 5 0 6 3 ^ 5 Pood Consumption The dally food Intake of the rats fed the 0.10, 0.03 and 0.01 percent diets was found to be from 10 to 15 grams per rat. It may be calculated that these rats, weighing about 200 grams, Ingested quantities of 2,4-dichlorophenoxyacetic acid of the order of 0.06, 0.02 and 0.006 gm./kgm./day, respectively. On the other hand, food consumption of the rats fed the 1.00 and 0.30 per cent diets was quite low, the exact amount eaten being uncertain because the animals wasted a great deal by pawing and scratching at the hoppers. Growth and Survival The rats on diets containing 1.00 and 0.30 percent 2,4-dichlorophenoxyacetlc acid lost weight rapidly after the start of the experiment (see chart) so that they were killed and examined after 12 days. One rat on the 0.30 percent diet died (Table I V ) . The rats receiving the diet containing 0.10 percent 2,4-dichlorophenoxyacetlc acid did not grow well. experimental period (113 d a y s ). Only one rat survived the full Two of the rats In this group were killed because of spontaneously occurring diseases (pneumonia and ear Infection), one was missing from the cage, and the fourth was found dead In the cage In such a condition that the cause of death could not be ascertained. The rats in the groups receiving 0.03 and 0.01 percent 2,4-dichlorophenoxyacetlc acid In the diet grew as well as the animals re­ ceiving the control ration. Deaths In these groups were attributable to either pneumonia or ear infection. Hematological values obtained on the blood of these rats after 106 days on the diets as well as those of the control group are summarized in Table V. within normal limits. All results were 5101 -8- TABLE IV SURVIVAL OP FEMALE RATS ON DIETS CONTAINING 2,4-DICHLOROPHENOXYACETIC ACID Number of Rats Surviving on Each Dietary Level 0.00 0.10 0.50 1.00 0.01 0.05 0 5 5 5 5 5 5 9 5 5 5 5 *a 5 12 5 5 5 5 4b 5b 20 5 5 5 50 5 5 5 4C 41 5 4 51 5 4d 5 4a 5 4 4e 62 4 5 4 3 69 4 5 3d 3 79. 90 4 5 3 2d 4 *d 3 2 100 4 4 3 2 113 4 4 2C lc 9 W S 0 S M O Q Days on Exnerlment a- dead In cage b- animals losing weight, sacrificed for hlstopathologlcal examination c- pneumonia d- ear Infection e- missing from cage 5102 H S ? 8 : -9- Gcneral Appearance The rats killed after 12 days on the 1.00 and 0.30 percent diets appeared thin and unkempt. Pat depots were depleted. ^ On the othercA O hand, the rat3 maintained for 113 days on the 0.10, 0.03 and 0.01 percentOO CO diets appeared to "be in good condition. ^ Body and Organ Weights A summary of the average final b ody weights, organ weights and blood urea-nitrogen is given in Table VI. Increased organ weights on the "grams per hundred grams" basis for the groups of rats receiving the 1 .00 , 0.30 and 0 .10 percent diets as compared with the controls was due entirely to decreased body weights. The body weights and organ (heart, liver, kidneys, spleen) weights of the rats on the 0.03 and 0 .0 1 percent diets were normal. TABLE V HEMATOLOGICAL VALUES F R O M FEMALE RATS MAINTAINED FOR 106 DAYS ON DIETS CONTAINING 2,4-DICHLOROPHENOXYACETIC A C I D Diff.Count(percent) freutr. Lymnh. Eosin. Rat Number Percent 2.4-D Erythrocytes (million/ cu.mm.) Hemoglobin (gm./lOOcc.) Leucoeytes (thousand/ cu.mm.) 8092 8076 0.0 9.0 14.4 14.0 29 68 3 0.0 9-3 14.2 21.7 14 78 8 7950 0.0 8 .1 14.0 13.5 - - 8099 0.0 8.4 14.1 13.4 20 71 9 7947 0.1 9.2 14.0 22.6 72 8079 0.1 9-.9 14.7 20.5 25 - 3 . - • 8101 0.03 9.0 15.1 18.7 12 83 5 8088 0.03 9.0 13 .6 24.0 12 82 6 - 5103 0CC2SÏ3 TABLE VI AVERAGE BODY WEIGHTS, ORGAN WEIGHTS A N D B L O O D UREA-NITROGEN VALUES P R O M GROUPS OP FEMALE RATS MAINTAINED ON DIETS CONTAINING 2,4-DICHLOROPHENOXYACETIC A C I D Percent In Diet Number of Rats Days on Diet Body Wt . (gnu) Initial Pinal* Organ Weights igm./lOO gm. body wt .) Spleen * liver Kidneys Heart ^ Blood Urea Nitrogen f 0.00 4 ' 113 174 206 0.40 2.44 0.73 0.33 29 .2 0 .0 1 4 113 175 207 0.39 2.33 0.72 0 .30 18.9 0.03 2 113 182 203 0.36 2.39 0.75 0.25 17.1 0 .10 1 113 176 175 0.38 3.14 0 .78 0.34 12.5 0.30 4 12 177 145 0.41 3. 5 7 O .89 0.39 37.7 1.0 0 5 12 175 127 0.39 4.72 O .85 0.47 2 0 .1 A Fasted overnight o t CJ o 8 ^ 8 8 0 S M O < 1 ro r- - o 1 0 1 * -11- DOW508349 Hlstopathology Microscopic examination of the tissues revealed a slight parenchymatous degeneration of the liver ceils In the rats on the 1.00 percent diet. Two of the four rats on the 0.30 percent diet showed slight cloudy swelling In the liver and In the proximal con­ voluted tubules of the kidneys. There was no evident Irritation of the stomach mucosa of the rats on the 1.0 0 and 0.30 percent dietary levels. The one rat surviving one the 0.10 percent diet showed only a very slight cloudy-swelling In the liver. None of the animals on the dietary levels of 0.03 and 0 .0 1 percent 2,4-dlchlorophenoxyacetic acid showed any changes upon hlstopathologlcal examination. SUMMARY A N D CONCLUSIONS Groups of 5 female rats each maintained for 113 days on diets containing 0 .00 , 0 .0 1 and 0.03 percent (0 , 100 , and 300 p.p.m.) 2,4-dichlorophenoxyacetic acid showed no adverse effects as Judged by growth, food consumption, general appearance, mortality, blood urea-nitrogen concentrations, hematological examination, organ weights, and gross and microscopic examination of the tissues. On the other hand, slight adverse effects were observed at concentrations of 0 .10 percent (1000 p.p.m.) fed In the diet for 133 days. Extreme weight loss of the rats on the 0.30 and 1 .0 percent (3000 and 10,000 p.p.m.) diets was associated with a low food Intake and necessitated sacri­ ficing the animals''on these dietary levels 12 days after feeding was begun. 5105 0 0 0 2 8 io • >4, 230 WEIGHT IN G R A M S 220 210 200 190 AVERAGE BODY 180 170 160 150 90î£ 140 0SC80S MOQ 5107 2 A . 5-T ACID po Trichlorophenoxy Acetic Ac1.d 93•93 * cn Trichlcrphenol 0 .7 6 % co Sodium Chloride 0,6% CO Freeze Point 150.6®C. Color Whits 2,^-L ACID Batch 7-2 cn / ’ 2,h-D 99.35% Dichlorcphenol 0 .1 Z % Sodium Chloride 0.035% Freeze Point 136,5°C. Color White 3 . & . S C . 2. 7 7 ¿j.. / 3 — L.. C . Reference . . r^dJC^L 1 3 . $ 4 5 . Sent.________ to ..•C at _____ m m . Hg De n s .---- at — ---- #C/_________ °C T 2 1 .___ BP. Handling Precautions Product state : Research .-.— — — Development .— — — __ Sp ec . Che m .--------- Plant------Use CONTEMPLATED---------------------------------------------------------------------------- --------------------------------- --------------------------------------------------------Tests requested : L» ^ * Y - Charge No . / / ■ * ? ■ , & u A A y - • * ¡ 1 ^ .1 1 . L it . Re f . — , D iscl. No .________ Patent ------------(Please do not write selow this line ) Tests By Date • • • • • • • • • a ••a a ta a a a a a a a a a a a a a a a « a a a « a a a M a a M a * ia a «a vt/ JA General screen i ng................—...........—Su!....—............ „ 7^:U7* Noted By__i. S a m pl e .1.datd.z!>zln.. .B y 5112 t o ___________________________ .........................D a t e ........... . CT C C c 'C P a r M. c IB. ASSOULY (Paris). , Le désir d'obtenir de belles récoltes a, de tous temps, été lé souci perm anent des agriculteurs. P our y parvenir, ils ont essayé divers moyens : enrichissement du sol p a r des engrais; accroissement du rendement p ar l'emploi de substances de croissance; cnlln, destruction des mauvaises herbes qui, en absorbant les engrais, l'eau et les matières minérales, entravent le développement des plantes utiles. On est parvenu aujourd’h u i,'d an s le domaine des produits de croissance et des herbicides, A des résultats tout-ù-fait remarquables. Mais leur fabrication suscite des inquiétudes dont le Conseil Supérieur ¿"Hygiène de France s'est fait l'écho dans sa séance du 13 février dernier. On a d ’abord pensé que ces produits pouvaient présenter une action cancérigène, et cette appréhension s'explique fort bien quand on voit là ... manière dont ces substances agissent sur les plantes, où elles provoquent des tum eurs, des hyperplasies. Mais flnalcmcnt, cette action cancérigène a été écartée p ar le pro­ fesseur Simonnet. P a r contre, le Conseil Supérieur d"Hygiène de France a souligné le danger d’intoxication, que pouvaient présenter ces produits pour les ouvriers qui les fabriquent. A vant de nous attacher à l’étude de la toxicité des substances en cause, nous croyons utile de rappeler quelques notions techniques à leur sujet. c o m p o s it io n e t a c t io n d e s h o r m o n e s v é g é t a l e s ■ Nous avons employé les mots d'hormones végétales, parce que c'est ainsi que l'on désigne couram m ent ces produits destinés à accroître le rendem ent. Il conviendrait peut-être d'employer plutôt (comme le remarque Pierre Chouard) le nom de substances de croissance qui désigne, sans préjuger de leur nature intim e, les agents chimiques de tous ordres, capables d'exercer, A de très petites doses, une stim ulation puissante sur l'élongation des cellules végétales jeunes. Primitivement, ces substances o n t été appelées auxines ou hormones de croissance végétales ou phyto-hormones. E n réalité, les auxines sont seulement les substances fabriquées p ar la plante ellemême e t qui exercent ce phénomène de stim ulation, d'élongation cellulaire p a r un . mécanisme horm onal.. On s'est aperçu, ces 10 dernières années, qu'une m ultitude d'autres substances avaient le même pouvoir que les auxines; on les a dénommées hormones végétales. . . . . Ces hormones ont une action qui varie suivant le moment d’utilisation et le b u t : recherché : elles exaltent la croissance des plantes ou, au contraire, la ralentissent; elles accélèrent la germination des graines, excitent la formation de racines; de germes, ettent .de —— boutons, de ----—fleurs; —» •»— ou, au contraire, —7 -ralentissent ces fonctions; — r elles —— — sperm -----------------—la formation artificielle de*fruits sans graines, sans fécondation p ar le pollen; elles aid en t (1) Communication à ia Sodili de Médecine du Travail, le 20 novembre 1950. 0 ( 3 1 2 LO 0 / 5116 I d D É S H E R B A N T S E T S U B S T A N C E S '¿ E ¿ R O IS S A N C E ' 27 . ; V FABRICATION DE L’ESTER DU 2-4 D " - wv L a fabrication de ces hormones végétales s'effectue seulement pen«lant quelques mois, en général de février à m ai, c'cst-à-Uirc pendant la période oii l'agriculteur a . ' besoin de ces p ro d u its.; •. n existe su r le m arché deux sortes de produits désherbants; l'u n est d base d ’e ster de l'acide 2-4 D. que l'o n émulsionne dans de l'huile minérale avec un m ouillant. L ’utili- ] ' sateu r procède d une seconde émulsion du produit dans l'eau et le projette ensuite |i 9 9 8 9 6 &la réparation des dégdtscausés p ar la gelée, facilitent la transplantation, le bouturage ' C’est dire que leu r champ d'application est vaste. • . Que 30nt hormones T — Ce sont .des dérivés oxy-acétiqucs du noyau benzine . e t du noyau n ap h talin e; des acides pliényl, naphtyl et indol-acétiquc, proplonlque et =■ . ■butyrique, ainsi que certains de leurs dérivés, comme p ar exemple des nitrilcs, des-. esters alkyliques, des sels d ’alcali e t d'ainmonium, e tc .- L 'un des plus connus est.. ■' l'acide 2-4 dichlorophénoxyacétlque, qu’on appelle, par abréviation, le 2-4 D. * Chacun de ces produits a une-action particulière et leur emploi varie donc suivant ■ le résultat & obtenir ; p ar exemple, pour le bouturage, on utilise l'acide indol-acétlque, ' '• l'acide.indol-butyrique ou l'acide naphtaline-acétique plutôt que le 2-4 D. qui a une 1 . " - action tro p déform ante dans ce cas; contre la chute des fruits, on se sert de l'acide naphtaline-acétique ou de scs sels, ou bien encore du 2-4 O . mql« à grande dilution. Signalons aussi que le 2-4 D. appliqué en pulvérisations ou aérosols, stimule puissam­ m ent la m aturation des fruits. - Un nom bre considérable de corps ont été ainsi étudiés et mis au point e t certains . " (comme le 2-4 D . dont nous parlons plus h au t) se sont révélés être de l’ordre d'une centaine de fois plus actifs que l'aûxine proprem ent dite; si bien qu'aux doses ordinaires ■ ce n 'est plus le pouvoir normal de croissance qui s’exerce, mois l'excès de ce pouvoir, aboutissant & la toxicité. De cette toxicité a dérivé un emploi nouveau : l’utilisation comme her bi ci de. . . . . . . . . . Cette utilisation des hormones comme herbicides sélectifs est venue aussi de la con­ naissance de leur action, stim ulante sur les diverses fonctions de la plante, y compris la form ation de tum eurs. L'usage des hormones, en ta n t que substances de croissance, provoque souvent, après l'application, des hyperplasies, des cals cicatriciels qui se form ent su r la surface de section d 'u n organe, tige ou pétiole en généraL Les hormones, -. à doses herbicides, ont une action intensifiée; elles provoquent une exaltation folle - des m anifestations de croissance p a r hypertrophie e t hyperplasie, une activité désor. donnée qui entraîne l'apparition de monstruosités les plus diverses, la consommation . ■ Intense «les réserves et finalement la m ort, au stade herbacé. n s'ag it donc d 'u n e destruction radicale, puisqu'il n 'y a pas formation de graines. Cette action est, en outré, sélective; les hormones s’attaq u en t aux mauvaises h erb es" mais respectent les plantes utiles. E t ceci s'explique facilement : la résistance des ' plantes varie su iv an t les espèces; celle des céréales, p ar exemple, est beaucoup plus . forte «pie celle des ravenelles, coijucllcots ou bleuets. C'est pourquoi lo traitem ent est - sans effet su r les céréales, d condition cependant qu'elles aient dépassé le stade 3 feuilles. Il devient aisé de composer un produit de telle manière «pie seules les mauvaises herbes soient touchées, tan dis «pus les autres plantes resteront insensibles d son action. Il convient, en outre, d ’observer, lors de l’emploi, certaines précautions pour obtenir . un bon résu ltat. E n général, on opère &la fiu-du printem ps, lorsque les tiges des plantes . o n t a tte in t leur plein développement et ont épuisé les réserves de leurs racines. On - vaporise le p ro d u it, en solution, sur le feuillage; il adhère fortem ent, pénètre rapide. : m ent dans les feuilles; la sève le véhicule ju squ'aux racines; il provoque alors, en même tem ps q u 'u n a rrê t de croissance au stade herbacé, une prolifération désordonnée des . cellules «pii a b o u tit & la m ort, ainsi que nous l’avons vu. L a destruction est obtenue en 15 jours ou 1 mois. " - Ces produits présentent un avantage considérable sur les désherbants habituellem ent • - employés : chlorate de soude, qui d étru it to u te végétation sans distinction; acide sulfuriipie, qui acidifie le terrain , ou sulfate de cuivre, susceptible d'une légère sélec-.Uvité, mai» d o n t la m anipulation présente do graves dangers ou des inconvénients e t ; > ■ ' d o n t l’action n ’e st pas - aussi totale. ; „ ■ -v ;: ;v " . H O.0 , s 28 M. A SSQ U LY llxé par le mouillant, adhère néanmoins. Le second qui semble avoir la faveur des • agriculteurs, ne contient ni émulsifiant, ni mouillant.. A base de sel de soude du 2*4 D. il est directement soluble dans l'eau. Mais il risque d’étre lavé par la pluie, puisqu'il * ne contient pas de mouillant. De nouveaux produits à base d'ester sont actuellement en fabrication, comprenant une plus grande quantité d'émulsiflants. On pourra alors verser le produit directe­ m ent dans l’eau. Le mélange se fera seul, sans qu’on a it même besoin de l'agiter. D ans^'usine que nous avons visitée, on procédait seulement & la réaction d'estéri­ fication de l'a d d e 2-4 D. qui arrive to u t prêt d'une usine de province. ; Ouvrons une parenthèse pour signaler que dans cette usine qui fabrique l'acide lui-même, les ouvriers sont imprégnés d ’une très forte odeur de phénol, caractéristique de ce produit. Cette odeur est extrêmement tenace; clic persiste, même lorsqu'ils q u itten t leurs vêtements de travail. Ce qui leur a occasionné quelques désagréments : on considère, par exemple, leur présence indésirable dans les lieux publics, cafés, tram w ays, etc.... Fermons ici notre parenthèse et revenons à notre usine parisienne. La réaction d'estérification a lieu à partir du sel de soude de l’acide 2-4 D. sur lequel on fait agir de l'alcool éthylique à 95*. porté è une tem pérature de 70*. A ce mélange, on ajoute ensuite de l'acide sulfurique à 98*. ce qui entraîne une élévation considérable de tem pérature. La cuve est alors refroidie et maintenue à 70*. Cette opération ayant lieu en vase clos, l'alcool distille et retombe dans la cuve. . Le produit est ensuite lavé à l'eau, toujours en vase clos et on procède h une décan­ tation. 11 reste alors dans la cuve un liquide sirupeux qui contient environ 1 p.- 100 d'alcool et qui dégage encore une forte odeur de phénol. PATHOLOGIE _.\V • .;;v L’Interrogatoire des ouvriers affectés & cette opération révèle qu’lis présentent les •. troubles suivants : somnolence avec sensation de jambes lourdes, irritation des voies aériennes supérieures, gastralgie avec perte d’appétit, goût sucré dans la bouche avec ■ ... . hypersallvation, sensation d'ébriété et d'hypersensibilité de l'ouïe, le moindre bruit faisant sursauter les ouvriers. Les ouvriers employés au conditionnement se plaignent d'asthénie avec jam bes . lourdes, sensation d'ébriété et de tête vide,. A quoi pourraient être dus les symptémes signalés ci-dessus ? On peut imaginer qu'ils sont causés par le produit Uni lui-même ou par les émanations des différents consti- * tu an ts, au cours de la fabrication. C’est ainsi que l’irritation des voles aériennes supérieures est bien connue dans l'Intoxication par les phénols, qui produit des catarrhes (laryngites, trachéites). D 'autre p art, il est signalé par les auteurs que l'intoxication chronique p ar le phénol — d ’ailleurs discutée — peut se m anifester entre autres p ar des troubles digestifs, ■- J anorexie, ptyalisme, céphalées, vertiges. Or, les ouvriers nous ont signalé tous ces • ' . signes, parmi les troubles qu'ils rattachent & l'exercice de leur profession. * • ■ : ' - On pourrait rattacher également quelques-uns des symptômes relevés aux émanations ■■ probables des vapeurs d'alcool en cours de fabrication. Cette opinion n 'est toutefois . pas partagée p ar l'Ingénieur que nous avons vu pendant notre visite, car, d'après lui, . d 'au tres fabrications m ettan t en jeu des quantités im portantes d'alcool ne présen-. ‘ tcralcnt pas les inconvénients signalés par lo personnel. ■ E n résumé, on peut donc dire que la plupart des signes pathologiques peuvent être dus au dégagement de vapeurs de phénol. Toutefois, des signes plus particuliers, tels que le goût sucré, la sensation d ’hypersensibilité de l'ouïe paraissent originaux e t • dus au produit fini lui-méme. Les troubles rapportés ci-dessus pourraient constituer un syndrome caractéristique du travail au contact d erp ro d u its en question, si d'autres auteurs les avaient signalés. Nous avons procédé à des recherches bibliographiques, mois nous n'avons pu trouver ' - de référence, du moins dans les sources que nous avons utilisées, concernant, leur •- . : action sur le personnel employé è la fabrication. . . . . • -, 0012192 5118 6889 S I ZH0 0 1 . sur les surfaces à désherber au moyen d'un pulvérisateur. En cas de pluie, le produit, 29 D ÉSH ERBAN TS E T SU B ST A N C E S DE CROISSANCE . ■ P a r contre, diverses publications signàlcnL tes travaux effectués sur leur toxicologie. U nous paraît intéressant de résumer ci-dessous les recherches effectuées dans les différents pays du monde. '•toxicologie : Le professeur R . Fabre, dans un rapport présenté aux Journées Médicales Interna­ tionales de Paris en 1937; signale les tum eurs qui se forment chez la plante autour .d’une piqûre d’hétéro-auxine, acide indol 3-ncétlque. U termine son rapport sur quelques questions pleines d ’Intérét : < Les auxincs végétales ont-elles une action sur le m éta­ bolisme ou la croissance d’un organisme animal ? Aucune expérience définitive n’a été tentée dans ce sens jusqu’ici. Mais il serait fort intéressant d’établir un parallèle . entre l’action encore bien discutée des hormones sexuelles sur le développement des plantes e t celle des auxines sur le développement des animaux. E t s’il y a action, sur quelle cellule de l’organisme porte-t-elle 7 A utant de question sans réponses, qui doivent ten ter physiologistes e t médecins. » De même, cette question do la toxicité du 2-4 D. pour les humains fu t posée lors de la deuxième réunion du N orth Central States Wced Contrâl Conférence. Quelques membres relatèrent alors des cas d’ingestions accidentelles de quantités variables, mais toujours sans altération apparente de la santé des accidentés. K raus fit savoir à. l'assemblée qu’il avait lui-méme absorbé quotidiennement 1/2 g de 2-4 D. p u r pendant 3 semaines, sans ressentir de désagrément. . Des expériences furent tentées â Bcltsvllle, afin d’éprouver la toxicité du 2-4 D. sur les anim aux. L a première consista à foire paître, pendant 15 jours, des vaches et des moutons dans un pâturage préalablem ent tra ité avec l’herbicide. On avait même doublé, la dose habituellem ent nécessaire à la destruction des mauvaises herbes. Les' anim aux furent ensuite examinés p ar des vétérinaires, et trouvés en excellente santé. Dans la seconde expérience, on fit absorber â une vache 5,5 g de 2-4 D. par jour, e t ce pendant 3 mois. L ’animal consomma facilement le grain d’alimentation avec lequel la substance chimique é ta it mélangée. Sa production laitière ne subit aucune baisse e t un veau, nourri entièrem ent de son la it pendant 1 mois, ne m ontra aucun symptôme de maladie. Une autopsio n’a pas révélé d ’effet apparent du 2-4 D. su r les différents organcs.de cette vache. M. K . B jom e t H . T . N orthen donnèrent pendant 4 semaines une dose quotidienne de 200 mg/kg à des poulets. Cela fu t sans effet, étais la dose de 765 mg/kg fu t mortelle. E . V. H1U, e t H . Carllsle, entreprirent des expériences sur 450 souris, 150 ra ts blancs, 125 cobayes, 70 lapins et 3 singes. Le 2-4 D. fu t administré avec les alim ents, en injec­ tions intra-veineuses, à des doses croissantes pour atteindre e t déterm iner la dose mortelle. A ce stade, les anim aux m eurent soudainement de fibHlatlon ventriculaire aiguë. S’ils échappent à la m ort instantanée, on voit apparattre la raideur des extrém ités, avec do l’Incoordination musculaire, la léthargie, la stupeur et finalement, le coma e t la m ort. ; Ces symptômes sont constants, quelle que soit la façon d’adm inistrer lo 2-4 D. e t quel que soit l’anim al utilisé pour l’expérience. . . .. : L’Investigation m ontre que le 2-4 D . e st-u n composé relativem ent peu toxique dont la dose mortelle e st de : ■. 375 mg/lcg d* souris, — de cobaye, ' 660 — • de rat, ' 800 ‘ ~ ’ de lapin, '-10 0 0 •-.* .• • ■’*; •/ \ • , ■: . . . ■•■ . „ ; quand ces doses sont administrées en solution aqueuse p ar la bouche. E n supposant que la tolérance pour l’homme soit similaire à celle des anim aux, la plus forte dose que pourrait supporter un homme de 75 kg serait de 15 g, ' Cés expériences o n t été conduites avec du 2-4 D. utilisé comme herbicide, softs que sa toxicité soit augmentée p a r celle de ses solvants. Aucune expérience n ’a été entreprise pour m ettre en évidence l’Intoxication chro­ nique avec les effets de l’absorption p ar la voie respiratoire. De leur côté, K lng et Prnfound firent des essais en aquarium sur la brème et le bar.Ils trouvèrent que dans ce milieu clos, le 2-4 D. â 1 p. 100 n ’est pas toxique pour ces poissons, mais qu'il le devient légèrement à 100 p. 100 , concentration qui ne peut être quo tem poraire et. locale en eau courante. ' - ■ ‘- E n vérité, on se trouve 1&devant un produit nouveau dont le mode d'action lui-même sur les végétaux n 'est pas encore très déflni en agronomie e t reste encore un peu m ys• térieux. —r — . . v. . E n ce qui concerne l'organisme humain, le problème est entier, au tan t du moins que .'n o s recherches. nous perm ettent de l'affirmer. • nûsuatû ■ ' ” ,r -.V ' 'f On a donné le nom d ’hormones végétales & certaines substances chimiques capables - < d'exercer, h de très petites doses, une stim ulation puissante sur l'élongation des celx Iules végétales jeunes, action exercée habituellement par une substance élaborée p a r . la plante elle-même. Ces hormones sont des dérivés oxy-ncétiques du noyau benzène . et du noyau naphtalène, des acides phényl, naphtyl et indol-acétique, propiontque *" et butyrique, ainsi que certains de leurs dérivés : nitriles, esters alkyllques, sels d'alcali, ' .. d'am m onium, etc_ L 'u n des plus connus est l'acide 2*4 dicblorophénoxyacétique, . appelé couramment 2?4 D. • . L'action intensive de ces hormones les a fait utiliser comme herbicides sélectifs. Elles agissent en provoquant une exaltation folle des manifestations de croissance, une activité désordonnée qui entraîne l’apparition des monstruosités les plus diverses, . la consommation intense des réserves, e t flnnlement la m ort au stade herbacé. -• Les ouvriers employés & la fabrication de l’ester du 2*4 D. se plaignent de somno­ lence avec sensation de jambes lourdes, d'irritation des voies aériennes supérieures, de gastralgie avec perte d'ap p étit, de goût sucré dons la bouche-avec hypersalivation, . de sensation d'ébriété et d'hypersensibilité de l'oufe, le moindre b ru it les faisant, sursauter. .. • - , Des expériences faites sur les anim aux ont montré que le 2-4 D. absorbé avec les aliments on inoculé en injections intra-veineuses, n 'é ta it toxique q u 'à doses assez élevées. ■ : am uopasm ix . . Bjorn , M. K. et N oirm sif, H. T. — Elfects of 2-4 Dlchlorophenoxy -ecétie acid on chieics ' Science, 29. 1948, p. 479-480. - - - B massb-Orossabo, L. — Les mauvaises herbes. Cuovaro, P. — Les progrès récents dans la connaissance et l'emploi des substances de crois­ sance. Revue Internationale de Botanique appliquée et d’Agrieullure tropicale, n“ 307,308; 309, 310. 311. 312,1948; n - 313. 314, 317, 318, 319. 320, 1949. - Cayouxttx, Richard. — Ce m erveilleux 2-4 D. Ministère de l'Agriculture, à Québec. F aurb, R . :— Les hormones végétales. Rapport présenté aux Journées ¿/éditâtes Internationales l&37f A iïitt MtTTr n Ki.r.. J . W ., Hoooso.v, R. E. et Oaetjkxb, C F . — Tolérance of farm »nimnt« to feed contnining 2-4 O. acid. J , Animal Science, S, 1946, p. 228. F am m fA, L. T. — 2-4 D. Indus. Hgq. Neunletter, 8,1918, p. 9. . l l u , B. Y. et Carlislx, H. — Toxicity of 2-4 O. for experimental animais. J . Indus. Ilgg, et Toxicol. 29. n* 2 , 1947, p. 85-05. J anot. — Phytohonnoncs. Conférence faite h la Aioiso.a de la Chimie, le 15 décembre 19-11. 0012194 5120 5121 IH 5122 T-C N PHENOXYACETIC ACID. 2,4n . DICHLORO Selective Herbicides and Growth Substances. Pathologic Effects on Man During the Manufacture of the Ester of 2,4-D. Arch. Mai. Profess. 12^, 26-30, (1951). M. Assouly. (French). Workers employed In the manufacture of an ester of 2,4-dlchlor phenoxyacetic acid complain of somnolescence w ith heaviness of the legs, Irritation of the upper respiratory passages, gastralgia with loss of appetite, of a sweet taste in the mouth with Increased salivation, a sensation of drunkenness, and hypersen­ sitivity of hearing, the least sound causing them to jump. In animal experimentation it was shown that whe n 2,4-D was Ingested Zift9Ç I 7»*nn ! or injected intravenously, high doses were required to produce intoxication. — Biol. Absts. Industrial Hygiene Digest Vol. 16, No. 4 - April 1952 0012195 5123 •Tndlreot Effects of Herbicides C. J. W i l l a r d Under ‘’Indirect Effects of Herbicides" wa have Included all eff e c t s other t h a n direct damage to the weeds or c rnz plants to w h i c h t n e y are.spoiled* The dis c u s s i o n will concern the newer o r g an i c h e r b i c i d e s only* This la an extensive U t r e r a t u r e on the effec t s o f sodium c h l orate and arsanicals, w h i c h does not need r e v i e w i n g here* 1* 2. 3* 4, 5* Eff e c t s out s i d e the treated field due to drift.or vapors of tne herbicides. Unfo r a e e n effecta o n ' t h e "balance of nature". D e s t r u c t i o n of wild flowers, ram© cover, etc., in r o a d s i d e spraying. P o i a o n o u a offocte of herbicides, &• Direct effecta b* P c i o c n l n g from poia o n o u a plants, not usually eaten, but e a t e n after spraying, c* P o i s o n i n g from o r d i n a r i l y harmless p l ants made p o i s o n o u s by tne effeot of sprays on tnem. E f f e c t s o n the c o m p o s i t i o n of crops. DOW 755434 T h e indirect effects.we have considered are: The first problem, drift of dust, droplets, or v«oore to adjacent or n o t - s o - a d j a c e n t s u s c e p t i b l e plants, has bee n so w i d e l y discussed that I will talcs time to m ake just one point about vapors. Any effe c t . o f vapors must be more or lees a mass action effect. Dr. Al b a n a n d T hav e U 9 e d exp e r i m e n t a l amounts of esters around susoe r t i b l e plan t s for years, w i t h no effects on them. But those wer e small plots. If y o u spray 10 acres with esters a n d wind moves slowly across this are a (Have you noticed h o w almost every story of vapor damage b e g i n s "There was h a r d l y any wind"?) the a i r may e a s i l y pic k up a t o x i c l o a d of vapor. As w e e d c o n t r o l men, w e . m u s t constantly emphasise the h a z a r d s Involved In u s i n g these e xtremely powerful chemicals, and p e r s o n a l l y and by precept, see to It. that every pr e c a u t ion Is taken a g a i n s t d a m a g e where no damage is intended. This oannot be said too often, o r made too emphatic. Carelessness in appli cation has a l r e a d y r e s u l t e d in some severely restrictive laws, and otners will f o l l o w if they seem to be needed. Several articles have a p p eared expressing tne fear that we "weed men" will run w i l d and e x terminate local flora of all kinds In all places. Ki l l i n g plants even with modern chemicals, still takes time a n d material, w h i c h cost money, «a are riot'likely to uso e i t h e r when we do not have c.n economic reason for doing so. In so d o i n g vp may kill some plants toat we do not ‘ntenl to M i l , simply b e c a u s e tnay are in bad comnanv, CGG5C13 5124 ■•■a Gtiould not be, nor >■ ; /.», n f- .i-J o r oblivious to changes In the b a l a n c e of should study .those c h a n t s r^nd, so far on poosiblo, avoid r e c o m m e n d a t i o n s thot produce unfr.vorr.blo changes, o r suggest ways of overcom.1 n?r them. For example, repeated roadside s p r a y i n g may take out l e g u m e s and, on many s^lls, reaulre nitrogen f e r t i l i s a t i o n t o - m a i n t a i n a healthy sod* UUW 755435 T h e c o m p l a i n t about k i l l i n g wil d flowers elong v * asides 1 g one to w h i c h >*a must, at least partially, plead guilty, ~ut without apologies* Home o f these complaints remind me of the nntlvivlsect l c r 'roups thr o u g h o u t the country, to whom, if one wore to Judea b y e i ther actions, the 11ft of a st^oy dnn> \p more aacrec than the lives of children, tfe wil l always hove some unreasonable c o m p l a i n t s , . So far ns In us ll9s, we should he "wise as serpents and h a r m l e s s as d o v e s “. We should listen to these complaints, *»t-udy them, a nd then, If the weight of evidence 13 then on the side of weed control, a n s w e r the m c o o peratively and court e o u s l y and (to ahead. Dr, E, P, S y l w e e t a r has done this b e a u t i f u l l y -In h1a roadside spraying campalffn in Iowa, He started tnls In cooperation wi t h the Highway Dore-traent In 19U£, The unsi g h t l y unremoved, deed br u s h a l o n g the road started c r l t ’cs ta l k i n g so vigorously that the c a m p a i g n was " o o f t - podalod" In l°4d, r e s u l t i n g In p o o r e r results and gre; ter e x p o n s g f o r the H i g h w a y Department, So, this spring, Dr. S ylvester arranged for a p r i n t e d d e bate between himself a n d o n e of his active o p p o n e n t s in the Sunday Dao Moines R e g i s t e r a n d Tribune, w n l c h has & Sunday circulation of nearly half a million. He aloo d e f e n d e d the r o a d s i d e spraying p r o g r a m at some 5° m e e tings t h r o u g h o u t the state, A.s a result, the Iowa roadalde spraying p r o g r a m is g o i n g ah e a d full steam, w i t h relatively little criticism. H i s h a n d l i n g o f the p r o g r a m was a model of public relations for all of us, 9o far as a n y t h i n g I might say her e d e f e n d i n g roadside spraying is concerned, I could o n i y say w h a t Dr, S ylwester said, 'or say it less veil. T hat art i c l e has been reprinted several times, most r e c e n t l y In "Down to E a r t h ” (Fall, 1950), I sue-gest that y o u get It. It is a t r e n c h a n t defense of the beauty, safety, and economy of g r a s a c o v e r c d roadsides, p r o d u c e d by spraying, a9 c o n t rasted with the miles o f p o i s o n ivy, w i l d carrot, brush, hemp, reftweed, etc,, etc,, w h i c h still corstitute the av e r a g e roadside. Folson! n£t by herbicides la n more serious matter, F o r c u n ? ‘r ely o u r p r o b l e m s are small compared tc those of the entomologists, but we defini t e l y do have problems. In the first place, any n e w thinsr gets blamed for a n y t h i n g that happens 'n its vicinity# 'vhen D", of o u r Department mad e a veeV*o tnu^ of ■northwestern.Ohio..in p u t t i n g on a few s p r a y i n g and d u s t i n g d e m o n e t ^ a c t l ^ ns wi fn ?, 4-D, two r e p o r t s of p o i s o n i n g followed - potn l a t e r sorwn to he °rrrneous, but i l l u s t r a t i n g tne tendenev to blame t r o u b l e s onto n n v f M n r new. Last s umm e r I I n v e s t i g a t e d s case of allege:! sheep nol soninr: by a CGQ'IRLS 5125 “b r u s h k i l l e r ” u s e d b y .a n •e l e c t r i c coop. Investi g a t i o n showed that there w a s t o r e a s o n a b l e chance that the arraying war. responsible, but the spray t r u c k p a s s e d alone; a l l t t l 9 before the sheep took sick, no the two t h ings wer e linked, bot h by the f a rmer a n d by the veterinarian. O f t h e n o w herbicides, T C A la caustic to the skin a nd the dlnltros a r e d e f i n i t e l y poleonoun. The Dii*s are a serious hazard to the p e r s o n a p p l y i n g t h e m — d e a t h has resulted from l ong e m o 3 u r e to a i r c o n t a m i n a t e d by droplets o f DN npray - but so for no co-clatnts of l i v e s t o c k p o i s o n i n g f r o m DN*s have come to our attention, and g r a z i n g e x p e r i m e n t a l l y sprayed ar e a s with them at four tires the r e c o m m e n d e d doses has g i v e n no serious results (3). The same is C true of T C A (£)• 5“ m T h e r o h a v e b e e n a f e w r e p o r t s of persons who are allergic _ to o r a f f e c t e d by 2 , 4-D. There is no r e ason to suppose that at least coma of t h e s e r e p o r t s e r e not true. These are i m p o rtant to '•'3 the f e w i n d i v i d u a l s concerned, and worthy of serious study, but we n o w h a v e e x p e r i e n c e enough to say that they are highly exceptional. ^ To hal o put the m a t t e r In p r o p e r perspective, in my p e r s o n a l ^ experience I k n o w mor e p e o n i e who are p o i s o n e d by e^vs then I have definitely h e a r d of b e i n g u n f a v o r a b l y affec t e d by 2,k-o, M a n y teste (£, 11, 1?) h a v e shown that stock w i l l consume grass s p r a y e d w i t h 2 , 4 -D a n d rea d i l y and w i t h o u t injury under o r d i n a r y conditions. T h e s e mate r i a l s h ave b een d i r e c t l y fed to p i l k c o w s wit h o u t I n j u r i o u s e ffects on the cattle. H o s t reports also state that t h e r o was no effect on the mil k - Dr, G r i g s b y in a l etter r e p o r t i n g u n p u b l i s h e d w o r k - t h i s summer ( l ^ O ) says that cows fed large q u a n t i t i e s o f 2,4— 3 gave m i l k w i t h a charactari sti c p h e n o l f l a v o r d u r i n g tho f e e d i n g and for 12 hours after the l^st cose of 2,4-1) h a d b e e n given. Th e s e e x p e r i me n t s and the experiences of thou s a nds of formers w i t h s p r a y e d p a s t u r e s p r e t t y w e l l dispose of direct 2,-’-0 toxicity to stock. However, almo s t all of ua have seen sprayed plants eaten that h a d not b e e n eater, b e f o r e - all the way from 9. M. B a l e i g h ’s case of r odents c h o o s i n g to eat rows of corn treated p r e - e n e r g e n c e w i t h 2,k-D to stock eating Canada thistles (0, Loe}, velvet leaf (J, L. Hutchison), Jimaon weeds (F. W. 9 1 1 f c ) t wild p a r s n i p (Geo. 3riggs, C, J. Willard), sunflowers (!, F. Yost), docks (Crafts a n d Harvey), r o u n d leaved m a l l o w (N. Shafer) and un p a l a t a b l e w e e d s in gen e r a l (Chas* J. Gilbert, II. S. Wood). Clearly, if any of these weeds were injurous or poisonous, their sudden c o n s u m p t i o n a f t e r s p r a y i n g could cause sickness or death of the stock. A c t u a l instances are few but suggestive, C. J. G i l b e r t m e n t i o n s oasee of p o isonous range weeds c a u sing death whe n eaten after spraying.' Bot h W a r r e n Shaw and O l i v e r Lee re>rort p o i s o n i n g of cattle from ? , ^ -D-crrnved wild cherry and if this O G O ielS 5126 - aIf r*t definitely p o i s o n o u s plant, la p r e s e n t in an area to be opraysd, it would c e r t a i n l y bo well to kee p stock out of it. J. K. F l eetwood reports h e a r i n g of a case of awl no eating* arrayed mature cockle-burs with severe sicknoss following* Crafts and Harvey (U) make a e l d l a r r e p o r t c o n c e r n i n g arrayed thiotles eaten by lambs* Virgil F r e e d m e n t i o n s oev e r a l inatances of ragwort c o l s o n l n g livestock - a f t e r b e i n g s prayed w i t h ?,U-D, He writes! "normally, l i vestock ^ b e c o m e p o i s o n e d on this p l a n t only when they are forced to graze it £ by lack of o t h e r vegetation, p a r t i c u l a r l y In the late winter and ^ early s o r i n g mon t h s i However, In the instances mentioned above, ^ it was a case of the a n imal a c t u a l l y seeking out this olant two orc. three days a f t e r s p r a y i n g a n d c o n s u m i n g sufficient quantities to.be ^ toxic. P l a n t s that h a d b e e n t r e a t e d for a o o r l o i l o n g e r than one to three day s d i d n o t a p p e a r to be pala t a b l e to .the animal and the initial p a l a t a b i l l t y w a s a t t r i b u t e d to th9 m a r k e d Increase in sugar that a c c o m p a n i e s a p p l i c a t i o n of ? , U - B to these plants* formally t he h i g h e r s u g a r con t e n t extends f rom F^.to ?2 hour« a f t e r spraying, t h e r e a f t e r d e c l i n i n g steadily," A wise precaution, therefore, would be to k eep stock out of s p r a y e d .'paaturas containing any poisonous weeds f o r 3 t o - d a y s to a week. M u c h the most s e r i o u s p o s s i b l e 2,4-D p o i s o n i n g effect that has bee n r e p o r t e d was due to the acc u m u l a t i o n of nitrates in sugar beet leaves t h a t hav e b e e n s p r a y e d w ith sub-l9thal amounts o f 2,^-0, For those d a t a I a m a l m o s t e n t irely Indebted to an u n p u b lished pa p e r submitted to "Science" by L* M, S t a b l e r and 2« iih!teheed, which I am a b s t r a c t i n g In part. N i t r a t e p o l o o n i n g of stock has been known for some time (1, 2, e s p e cially in c o n n e c t i o n w ith oat hay. Beet leaves from a M a n i t o b a f i e l d d a m a g e d by airplane spray i n g of a n adjacent whest field in l b b g c a u s e d the d o r t h of several steers, A local v e t e r i n a r i a n . r e c o g n i z e d t h e symptoms ae those of nitrate poisoning. T h e s t a ndard treatment, i n j e c t i o n with methylene blue, r e v i v e d l i v i n g but recu m b e n t st eers w i t h i n an hour. The leaves were analyzed '"for nitrate and showed n i t r a t e levels for above those known to be toxic. 5# 7) 0004816 In August, lQ^Q, acres of beets on seven f BT’rr.s In North D a k o t a w ere sprayed wit h insecticide c o n m i n ot'ei with it 19'ves fr'1m each fiel-3 and. flelds,' The lewvea frorr untreated fields a v e r a g e d 0 , ? ? p e r c e n t K?vO?; fhp. r. or the trod e>3 V*nge of 1,f ! l to ?.77 T)0rceut. matter of forgro 1a c n n«1.ie*,ed toxic. Is nr.t :i"ac’, but \ n "i■ .‘-•p tr, nltr1r.« v;-.1 cii n: 1i-• r* • a fi: t:1 Lun>., t.;V3 "•i¡i •i«) gur..;ood Cdrlnclell* fton, ) a nd opldortjort (T r a d o o c n n t l n a n r . ) hove also been indVroFel Tr^) na c o n t a i n i n g lstfcol auantiicio'a of nltrata under some conditions. Vi>i>436 Hr. C* D* Floyd, State Apiarist for Klnnocnta, reports one r a t h e r cloer ccae o ? p o l o o n l n r of bees p a s t u r i n g on mustard s p r ayed a f t o r it car.a I n bloom* Ho was not ablo to ducllc'-te the roault tfca nc^tt yoar, oven by f e e d i n g the boos oyruo contaminated by ouprooodly tho oomo 2 , b - D formulation used the y ear before. However, aor.^thlrr? k i l l e d the beaa the p r e c e d i n g y e a r (lntog}# Hr, Floyrl V o c o r t o of InJury wore r e c olved In 1^50. Apparently tho trou b l e is at loaat unusual. Tfcsre will bo many of these problems, reel and 1 mug! r.ary, ae w e e d control c o n t l n u o n to develop. C u r ettl.tu.1n, aa good weed control men, r.uat ba to c o n s i d e r every complaint, even those /r»i cl. lo o k absurd, cautiously o n d carefully, neither anylng "Impoedible" w i t hout investigation, o r fourfully a c c e n t i n g unproven r e a ronelblltty„ There h ave beer many reports of 2,to«o I n c r easing the protein content of ¡grain (c, ° t Ik), In gonb^al, there offoeta soon physiolo g i c a l l y to be l i k e the Increased p r o t e i n o b t a i n e d In a dry se&eoni that to, the p r o t e i n elements In the grain arc laid down first, e n d If the grain does not “fll*." veil, it. will b? higher In p r o t e i n than one that does fill* Usually, at leant, the p r o t M r p e r acre hr.a not b3«n I n c r e a s e d by spraying. However, m^ny o f t h o e f f e c t s on the v e g e t a t i v e p a r t s o f p l c n t e ( 1 0 , 3) a r e not. so s im p le and r v l 3o f i e l d i s open f o r the s t u d y o f th e e f f e c t s r f '2 , k —n t r . 1 rel*-t.pd compounds%on f o r n r e . The v e g e t a b l e r.en >now w e l l t h e -Trustor development o f f i b e r lr: rsr^rsp-uo, e t c , , f o l l o w i n g 2 , to—1> t r r r i t n e n t s * r n f i e l d c^^rp wo have no 1i>+s s 9 y e t , but th o problem l r hi c h i y 1 rro"t«..nt. 0004517 5128 < - 6 - l i t o r a t u r n Cited nr-adloy, \l9 B ai Crpcan, H. F. 5? Ponth, O. A* Ltveetnck p o l o o n ir.% b y .es« ¿127 a n d b t b n r p l n n t n c o n t aining nitrate, '/yominar Cook, B. consent tho a f fect of aoll tyro and fartliltora on nitrate L* of .tics acs Zita ospron*?c«l ono and the total nitrogen content of of t h o cr n l l grcrtnn. m . 2 l°ko. #v i As?r. Srp. S t a t i o n Sul. £*!•• J. Aa. soc« f.raron 2?: ■?r,7» lio? t (1OT0) 3» Corna, W. G. B f f o e t » of 2 , U » D a n d «jotl n o i nture on the catalase aatl v i t v , roapivatlon, a n d p r o t e i n content of bea n nlunta. Con. Jour. Poa. 2 ^ : 3 9 3~u0 5 ( l q W k. G r a f t o # ’-a . 8* avA Barvoy, Agren. l i 2 0 o * 3 2 C ( 1 9 ^ ° ) 5* ravidaon# tf. I?.# at nl* Nit r a t e polaontnrr of livestock. Can. Jour, of C o s p .t!od. Vol. 5, l^Ui* it* A* tteed Control* o o f , o n 1. Advancon In 6 . Srlclrcon# I. C.„ Sooly# C. I.# end Vlncoa# K. H. Cffoct of 2 , U - D uron o r o t o i n c o n t e n t of vheflt3. Jour. Aner* Boo. Arron. ^ 0 : (j 5 ^ ® 5 < 3 0 U . ^ 3 > 7. Gilbert# C. ^^roon, B. F,# Srndley, V, 9, A Reoth, 0, A, ’ Jltrstc? accu^nlr.tion in c u l t i v a t e d p l n n t a anfl weoda. 'Wyo. A p t . B*r?, station#; B a ll, 2 7 7 # 1 9 U6 . g. Grlpoby# 9, R# ft Fnpwcll# B, 1), Bora» e ffect» of h e r b i cide» on m a t u r e and o n (rraRins? ltvastoek. Rich. A?r, Uxo, station ^uavt, sail. Vol* 32# Ko. 3# Fob. l ^ O * Q. xV UQlu’OGoiij ¿j. i*. T b o offcct o f 2,&-D on vrhoat. Proc* -th Ann. M o o t i n g Fo r t h C o n t r o l Wood Control Conference# Bee* l^^f (Topol?n# ICnnaao) op. 7 7 * 3 ?• 10. H u l l 1near# C. H# Eff e c t » o f 2,* -I? o n the n l t r o j a n ond c a rbohydrate o c t a b o l l a a o f tho c o r n Rea* Boo. N.C.W.C.C«# Sprlnarfleld, 111# 1PU£, Bee. VIII# A t t r a c t 5, 11« Kaphart, L. «« M o d e r n t o r of discussion at f>t* i«ul meotlnc* of N.C.’ a .C.C# Proc. 2nd Ann. M e e t i n g »J.C.W.C.O.# at. Paul, Finn. 1<*5. PP. bg-75. 12. Mitchell, J, v.# et al* Tolerance of farm animals--to feed cen­ tal nl nff c# ^ - D acid. Jour. Anii.% r,c \ . 13* Olaon, C, E, * Dakota plants. Vhltahead. «•« ‘» i ^ r a t e content or Ac a . of 'Jci. Pf'J0,r 30 m e r.«ourh 1C-.1 ( i oitr-) 5129 • 7 " C.»: csr/J-.iS o f U îlla r â , 0 , J# Cífoofc o f 2#ÍM> on o ro tein noocarcH iioport II.C.U.C.C. Reooaroh Cocual tte e 77? Ctbc~3 'O 'j nono only aro from unoubltchod co-munlcntIona •to t*I:o vv USoï** . . . CG04E13 ,B ia n n j? , : Ja ü4- u f c o î a o f 2 , U-D on ancrai» boato. C ryatal-lzed fdOwG p-D’ïa oO';o.r b eota. V ol. XV, Ho. 1, Jon. 1959* n r r. r* V» * 15» &£ íjCLÜ . . •> ', . R-& D REPORT DOW C H E M IC A L U.S:A. » •• ES-274 RESTRICTED: lor ul« «««hin Th« Dow Ch«micol Company only. March 16, 1979 " t wfcN' '<"•.*• " Environmental Sciences Research 1, 9 1¡ 9, 0 ; 0 0. 0 0 3 • • _£ of " t ox ic i ty e l e v e n h e r b i c i d e s t o 20 d a p h n i d s ;.3£3Q9' ac ui a s m , ? AZ E ;»:«■ P j l : *-;h i 3 W. Z M. M c C a r t y $ s c ’• * r f»e s U //O / ' / ■ /*; ¿ “ fc^ t n e s s i j r i p T j A t .^ This report 1 s: 1 1 i nteri m [_X] ne* /J.O 4/c■ /-?<> DESCRIPTIVE SUMMARY WITH CONCLUSIONS: Hnclua« m ?hn space references *o 3o?a books, and ro earner re«j*C'j '•»<” is, potems and publications.; a n d mct nl y: n 1 Xj =!NAL E l e v e n h e r b i c i d e s w e r e e v a l u a t e d for a c u t e t o x i c i t y to the aquatic invertebrate, Daohnia magna Straus. T h i s s t u d y wa-s c o n d u c t e d to p r o c u r e e n v i r o n m e n t a l d a t a o n these pr o d u c t s . • i i 1 The the 48-hour sample's LC5O'values and their confidence evaluated are as follows: • • intervals for 48-Hour LC50 S atipie i . i• 2,4-Dichlorophenoxy Butyl ester DOWAIIOL* 8 <4 4(11 i Ester of2, 4 - D ^OWAIIOL PI3 Ester o f - ’2 , 4 , 5 - T •* ' ;• -*hleC 4 Formula 40 n R - c r b i.tfn DMA 6 K - 114 5 7 MCP amine r - n 2 .2 - ' ’ "■ PI3 . •'. : • ••' DOWAIIOL ethanol E3 ester Confidence ( 0 . 7 - 1 ' . 6 ) ’. m g / L 13*. 2 (.9.9-16,-5)' m g / L 13".7 .456 ester of of (0..00-33.5) ( 394-528)' mg/L mg/L C4 2 5 - 5 0 6 ) ’ m g / L 146 (36-223) 2,4-D ‘T r a d e m a r k mg/L • . 467 dichlorprop Interval mg/L •1.2 4'6. ( 22-r.ôè) Unseq. 3utoxy 951 2.,'4-D (248-2Î6)-t m q / L ’■•'iO. 6 ’ .(3-1*4) . .. DOWANOL Iso-octyl •1262 acid PIB;E3*te'r" of. S i l v e x T .o c S 'l’ DR.#lcf-«4/1 2,.4-D.'\ acetic mg/L 14.7 (11.7-19.2) mg/L 32.6 •'25.6-43.9) mg/L The Dow Chemical ■Li DISTRIBUTION? Ï ■* ,r i = - 1L £ i p 1 2 a j n * » a 7:C.*« X£ n *3 a l REPORT «NOÈ < s i r 1** ; • • ; r - - -4 ^ C604S2Q , 5131 755911 a w * mi¡ p ac 5132 t HN073280 l i e d } in United ätaU» o f America Bnrist»! fra» AssoaraoM cw Fooa it Dnoa Otkcims erras Ucnco Crttct Vol. XV, N'.. t, Ottnbtr ItSI CHEMICALS IN FOCÜ3: A IIEPOIIT TO THE ASSOCIATION OF FOOD AND DRUG OFFICIALS ON CURRENT DEVELOPMENTS. PART II. PESTICIDES ARNOLD J . LEHMAN, M.D. . Chief, Dleition of Pharmacology, V. B. Pood and Drug Adminitiraiion DOW'j95G 2 INTRODUCTION Tho pesticides bs discussed under the sectioned headings of acute toxicity, dermal toxicity, subacute and chronic toxicity, biochemistry, end pathology. In the tabutar eununarics to follow it will bo necessary to mention many of tho individual pesticides a number of times. Since their chemical names arc too cumbersome for this purpose ths common or short* term designation will bs given preference. A list of tho pesticides to be considered is given below. In this listing the short term is given firet with the raoro customary chemical designation, wherever applicable, presented in tho second column. Because of the diversified character of tho pesticides from both the chemical and utility standpoint, a classification based upon either one or the other of these aspects did not appear to be practical; hence, in an attempt to cegrcgato tho compounds into some useful order an arrangement was devised which is based partially upon chemical similarity and partially upon use. Chemical names will not bo repeated in tabulations and the groupings as presented below will be maintained. P lant Product! and Derivatioae 1 . Dorris 2. Rotations 3. Pyrethrins 4. Adothrin 3. Nicotino 0. Sobadiila 7. Jlyonlo Rotonono and rotenoids Oleoresins Allyi cinorin Vcratrino-Iiiio aiknioido Alkaloids: ryanodine, rynoino, and others 8. Dihydrorotonono Thtocyanatea 9. Th&nito 10. Lethauo-C0 11. Lotbano-334 12. Lelhano flpecisi Isobornyl-thiocynno acctato Uota-ihiocyano-cthyl esters o t aliphatic acids with 10-18 carbon atoms Ecta-butoxy-heta-thiocyano-dicthyl ether Lcthanc-00,3 parts and Lcthano-331,1 part Copper 13. Copper chloride 14. Copper carbonate 16. Coppor sulfnto hfer-ury 10 . Calomel 17. Corrooivo sublimate Merci. us chlorido Mercuric 'hiorida 0GG4S4C 122 5133 . ■•m - «. .. I S v~t»< y _ir ASSOCIATION OV FOOD AND DRNQ 0 F 71CTAL0 123 18. E thyl msrcurio pbozphato 19. Thonyl mercurio triothanol ammonium lactate Arsenic 29. Arcenio trioxide 2 1 . Potassium oroanito 22. Paris croon 23. Calcium nreonnte 24. Load areauato Coppor acato-arsenite Fluoride» 28. Dariuia fluoailleato 23. Cryolito 27. Sodium fluorido 23. Sodium fluosilicato / Sodium aluminum fluorido o O Afiacellaneout Mclala 29. Sodium eelonito or cclonato 29. Cadmium chlorida 31. T a rta r ematio * Potassium antimony! tartrnto -3 32. D D T 33. T D E 34. D FD T 35. Mothoxychlor 30. TI11I 37. A im 33. n m i 39. n u n 40. Lindauo 41. Toxaphsno 42. Chlordano 43. Aldrin 44. Dioldrin 45. Iloptachlor 48. Prolan 47. Bulan Dlchloro-diphcnyl-lrichloroothano Dlchloro-diphonyl-dichloroethana Difluoro-diphonyl-trichloroothnno Dlmothoxy-diplionyl-trichloronthana Technical bon zone hesaehlorids Alpha ¡corner of bcnznno hoxnchlorido Bota lsomor of bensenn hexochioride D elta isomer of benzeno hasachlorida Gamma iaomer of bensono hoxacblorida Chlorinated campbcna l,2,4,0,0,7,fl,8-Octochloro-2,3,3a,4,7,7a-liexohydro-4,7-math,-moindece 1,2,3,4,10,10 • ücxachloro • 1 , 4 , 40 , 8 , 8 ,8 1 • hoiahydro-l,4,8,8-d:m thanonapbtiialone 1,2,3,4,10,10 • Hoxschioro • 0,7 • epoxy • 1,4,40,5,5,7,8,8a - ootahydro • 1,4,5,8 • dlmothnnonaphthalona 1,4,5,0,7, 8 ,8 • llcptachloro - 3a,4,7,7a • tcirahydro-4,7-metiianoindono 2 -H itro-l, 1 .bia(p-chlorophcnyi) propane 2-Nitro-l,l-biu(')-chloropl)snyl) Lutano Organic Pknpkaiia 48. Parathiou 49. D im ethyl paratiiion CO. Paraozone 51. OMPA 82. EPN C3. T E P P 54. III5TP 55. AB1M7 0,0-Diotbyl-p.nitrnphepyl-thiophwiphatrj 0,0'D itocthyl'p.iiitrophr.nyl-thiophoephale 0 ,O.Diothyl-p-r beta • acetyl • ethyl • 4 hydroxy-coumarin Strychnine nitrato 72. 1030 73. Castrix 74. ANTU 75. W arfarin 76. Strychnine 77. Thallium oulfato 78. Usd cquill 79. Zinc phosphide Miteellantoun 80. Phygon 81. Spergon 82. Phsnothiazine 83. Aramito 2,3-Dichloronaphthaquionno Tctrachlorobcnznquinone Thiodiplionylamins B eta' - chloristhyl - beta - (p - tert. - butyl • phenoxy) -alpha -me thy 1-cl hy 1 r.ul fit c Tctrainctliyl-thuirum-disulfide Glyoxalidino acetate Zinc and r.odium iedtn of ethvlcnc-bia-dithiocarhr.mic acid 84. Arasan 85. Fungicide 311 C SO. Dithar.c S E C T IO N I . A C U T E O IIA L T O X IC IT Y The acute toxic dose valuer for the pesticides arc listed in Table I. Practically all of the data were obtained in the laboratories of the Division CG04C4S 5135 ASSOCIATION OF FOOD AND DUUG OFFICIALS 125 of Pharmacology, and, with a very few exceptions, the rat was employed as the test animal. The values for a few of the older compounds ouch as derrta, sodium fluoride, and arsenic trioxido cro so well documented in the literature that they were not necessarily checked in thb laboratory. In some instances technical difficulties would not permit the calculation of „ an LD hi hence, in the interest of accuracy tho term “approximate LD»” ia given as the column heading. Wherever possible ths pure compound dissolved in an innocuous solvont was employed, and dosages were ad* ministered by stomach tube to fasted animals. In an attempt to present as much useful information as possible in tabular form, the chief characteristics of the poisoning symptoms, tho time elements involved in their appearance and duration, end the clinical cource of the poisoning as observed in the onimal3 are briefly summarized. A few additional comments on tho various groups of pesticides follow. Thiocyanaiea: Except for thanite, tho acute toxicity was determined on the commercially available concentrates which aro 50% solutions in dec* domed kerosene. The LDeo’s as tabulated were calculated on the basis of the pure compound. Copper: Although the compounds listed are not used as pesticides, their toxicity doss ccrvo to give eome estimation of tbs toxicity of copper oxy* chloride, oxide, phosphate, silicate, zeolite, oxychloridcsulfatc, and Bor* deaux Mixture. Mercury: The general trend of phenyl mercuric compounds is an J,D»o of 50 mg.A s- or less. Arsenic: The ratio of toxicity of arsenites to arsenates may be considered cs 1:1.6; hence, other arsenical pesticides not listed, such os sodium and zinc arsenites, would have an LD*a of about 13-14 mg./kg., and copper, magnesium, and manganese arsenates would be in tho 22-23 mg./kg. range. Miscellaneous Metals: The value for sodium selenite or sclcnate is calcu­ lated on the basis of selenium. The toxic dose of cadmium chloride is listed because this metal is a constituent of a t least one fungicide, a com* bination of cadmium copper zinc chromate and calcium sulfate. o •a to ci CÍ CONTINUED IN THE N E X T ISSU E 5136 t A m t s t.-J II: Lily c ï ¿c A rra '"« * u v .3 i U à d -J G ^ is ìn ii li» (hr:t f C ^ rjU ra DOW75S5G! C . a h d C w :::a r i . i Natural producía cad drrivctiras 1. Dorris -1CS1 Frequent c'cu7ubivo Within minutes 1 to 3 days crisuraa followed by to fi to 0 hours drprczsica cf central errvous cyctsa 3. Rotonona 153 Caca sa for ¿arria 3. Pyrothrins £33 Central norvous eyatom Usually within 10 hours irritation; hypsrascit§ hour ability, trasera, and convulsions Death may bo delayed for as long as 8 day?, respiratory paralysis is a major cuuts of death 4. Allothrin C3> Tremors and canvulaiona COminutes Fatalities nppesr to b# rare 21 hours after poisoning. Death due to respiratory paralysis 5. Nicotine CO-CO Clonlo convulsions Within minutes 8 to 20 minutes Curaro-liko psralyds of respiratory mus­ cles and death 0. Echadilla teso Hatching, nuasubr eparms, otada, coma Within S min□tC3 7. R yania 780 Depression ef central ner­ 6 to 8 minutes vous oyctem 8 . Dihydrorotonons CO 0 bosra Death by respiratory failurs in 21 hours. Fatalities ram after Crd day although death c a y fcs delayed 10 cr 12 day»' 2-1 hours Gymptomorimllar to aeenito. Death from respiratory or cardiao paralysis may occur within 3 to 10 minutes after tha ingestion m a fatal dess 2 to 10 hours Continuous depression up to term inal convulsions. D eath usually within 24 hours although largo dccce m ay ba fatal within the hour after ingestion Rapid callapcs within min- Occasionally oymptoms m ay be delayed for a few hours. D eath isuus to respiratory paralyîis es for dsrna Thiocyanates 0 . Thanlto 10 . L othsns -00 1 1 . Lcthano-ESl 12. Lethono Special 1000 6G0 CO 400 Deep depreoeion, cyano- Within a few minutes cis, dyopnes and tonio convulsiona UtC3 Copper 13. Copper chlorids _ 14. Copper carbonate 18. Coppar sulfate 140 180 SCO Violent retching, rcuscu- Within a few minutes lar epaems end collapse 1C. Calomel If dors is ratai nod could bo fatal in 1 hour Symptomo of gastrointestinal irritation may cubsido, b u t dcath may foiluw csvcral daya inter bcccuss of damage to im portant organe ouch as livcr and kidnsys Mercury Gyrapt oms of morcurialism ut tliis dore b u t no desths (210 ) 17. Corrosive cublim ita 37 S h eet, dyeautsry, and anuria 13. E thyl mercuric phewpkata 19. Phenyl mercuric tricthunoi ammonium lactate £3 Similar to corrosivo cublim ate 30 l i Immédiats Symptoms nernet till death Shoak clTcct oí lits irritan t may be fatal within 21 honre. Acuta kidnay i la rangs m.By ho fatal by iho tlîird dey C0Ü'Îg4 ? 5137 ARSENIC 20. Aresuio trióxido 21. P otom iun amenito 32. Pttria_grc3n 23. Caloium nrconato 21. Lead arssn sta 13 M £3 Violent c ^ trc o n to ri'ia, dinrrkca, rico water stools Almost immediately A fow hours to several days £0 ICO Death from exhaustion and dehydration. Convulsions and general paralysis may oesur before onset of B.-u.trccnterilie. D eath -within -a-fow hours to several days Fluorides 2fi. Barium flaosilicato 23. Cryolite 27. Sodium fluorido 23. Sodium fluosilicate 175 2C9 200 125 Abdominal diotrers, diar- Immediately and ur. to 0 rhea, cyanosis, dye?* non, fibrillation of skclhoura ota! rouaclea «* Stupor and woakness . may persist for 23 hours In fatal poisonings death can result in a fow minutes 12 to 18 hours D eath duo to respiratory failure Miccolliu'couj Metala Garlio breath, nervousness, central nervous system depression V/ithiu 15 min­ utes 83 Salivation, diarrhea 15 to £0'm in­ utes 115 Percllolo thoss given for aresnio poisoning 20. Sodium eolenito or eelensto 2.5 30. Cadmium chloride 31. T a rta r emetto vomiting, i G astroenteritis may be a contributing eauso of death. D eath within a few hours to several daye C C c t Chlorinated hydrocarbons 32. D D T 2 S0 Extreme excitability, trem ors, twitching, convulsions, eoma, death W ithin com m ­ utes 2 1 hours Time interval for a fatal outcome is var­ iable, but death usually occurs within 2 1 houra 33. T D E 3100 Lethargy W ithin 21 hours 2 to 1 days Convulsione do not appear to ko a major symptom. If death has not occurred by the 1 th day prognosis appears to be favorable 3-1. D FD T 1120 Similar to D D T 35. Mcthcxychlor C000 Largely depression of centra! nervous system Within 21 houro 2 to i days Tremors havo b/.en noted but tr s not a prominent symptom 30. TB II eco Convulsions which may givo W3 y to a central nervous sjviem depres­ sion Within com m ­ utes 2 weeks Symptoms may persist for 2 weeks and deaths have bean observed lata in tho 2 wsoko observation 37. ABH eco H ypercxeitability convulsions About 1 hour 2 weeks Deaths delayed up to 2 weeks with con­ vulsions pcf.aistin* to the cr.d 53. BBH coco Principally tremors 2 to b hours 2 dayj I'jflth occurs between tho i’:id and day 30. L'J-il KvO Depressant to the cent ml nervous system 1 to 2 hours 1 day» Death may Le delayed to between liu: 2nd And Olh il.-i.y j ^ ~ ^ Apparently the f.uurico part, of tho mole­ cule does not contributo to tho sym p­ toms of ecuto poiconing end • 513 8_ - •; < V j! lililìaO lLj'J—ii/ 1‘ith.ft L. tr/O-ilvL? |.C: idle? l a ^ i k a . Couvalncsa pin. tn th n iu d IlyFSK-aaíUvlty, trem­ 1J bou» erai end convulsiona 21 bou» bloat of tho fatalities occur within bourn. Dels e i dratha have bean no' uptotLoCihda 7 457 Hypsieresltivity, trcm< era, end convulsiona 16 minutas 2 ¿ayo Treeless brgin in lOminutesjcouvnloio' appear efts? about 21 hours end tn persist for 2 days; deaths may bo c layed up ta tho Cih day 43. Aldrin 07 Trcmc-m ned convulsiona Within 1 hour 2 ¿aya Tims of death variable; may bo delay up to tha 6th day 44. Jloldrin 87 Tremo», muscular epanres, ccnvukions Usually within 2 days tho hour Ccatha may ba delayed up to tha Cth d 07 longer 45. Hcptachlcr C9 Tremo» and convulsiona SO minutes to on hour 2 days Clmilar to aldrin, dieldrin end calorda 49. Lítidans 125 l7yp:tt:n.¡UivjSy convukiuns 41. ToxcpLsns es 43. Chiodano, technical and 13 ióísuísj i»* u 45. Proion 4000 Tremo» end eonvuldoco Within about 1 Prolonged for Persistant convulsions up' to time dcatn which may not occur within days hour houm 47. Bulan 3S9 Convulsiona About 2 hours Fatalities uaually occur within 4 h ^ 4 bou» 5139 GGOS343 cr cc v Organio Phosphates 43. Parethion 49. Dimethyl parethion 49. Parsozono 61. OMPA S3. EPI7 63. T15?.? 64. TOTIP 63. AOP-47 63. E-003 3 16.2 7.6 13.6 14.5 1.3 7 6 19 Generaliced fibrillary tremo»i ealivetion, lecrimation, diarrhea end convulsions • lGccinutea to 1 12 hours hour Tha symptomatology cf tha individua organic phosphates iaeimilar. Dcatfc usually occur in 1 to 21 hours Dic¡t¿ o Compounds 67. DNOC 63. DN-111 23 ZZO Increased respirator}' rato Within hour tho 21 hou» Time of death variable hut usuali within 21 hours with byperpyrcai contributing causo Several h o u » D eath occurs G overn! hours after ¡exes tion Lesa then 21 hours Most fatalities occur within tho first hours Herbicides £9. 2,4-D £C0 A taxia, m yctcria, gas­ 29 minutes trointestinal irritation CO. 2 ,4 ,6-T 220 Similar to 2,4-D 61. Naphthaicno acctio ncid 62. M ctbyl-1-naphthalone acetic acid 1053 G astroenteritis, depres­ sion arid paraiyeia CO minutes acra Lintlcccucrs W ithin m inute: 21 Lourd TX Tr«* .•»»«*» îin.-l ra the noia 85. D ithane 5GG3 |: D iarrhea Delayed dentbs over 1 wce'x 3 dayo 3 hours Ba­ Most deaths occur within 5 days Delayed deaths up to 2 wcakj 1 1 5141 223961 Parto«« bO ■ --vllfel *' V-.-;VS?á topaala 00« !•■■««■« U • 4 *0 3 # ^ a plooo ■ f o n o« «os fo o « . n a atibada eaoaaBM tfela r o M lly , t t w a r o U lt s Ufe M tin ta « tram a tba atemafe tafea, ta ita a aaa*. fiarlas tfea ata«? tfea h M I j n r t «fei| ¿ o str a l bloc« eouata aara tafeaa b a fw * «fea «áfelU atrtt 1 aa 1 toa «ru«» °a U a J-Hfe «ay tac aa tba Ota «ay «t taa toadla t i t a cf tfea I t ataa^JiCblfe aaa «abarato*« atut a p b etoalactrta col «Timbar ( 6 ) . laatb a* « t tba e ca p la lta a of tba atudy, taa a alaala 1 Qpafe tfea aataaM , tu a a a a lataatifefe . . lúa«, s« « rt, llv a .-, kldaay, adraaai, a p la ta , tfeyroU ^ M »r¿ •I • “*31* wrary *■ taataa acd flxa« ta líí foraalla. í «ara takaa froa taa « a. Jyajteaa jruduatd. m Tba a ffa a t of m rlou* «oaaa o« 2,4*0 «a eurvival for 1« «aya ta aojan ta rabia 1. Iba oral IB^ q 1« appraxlaatal? 1>J aa/kicx. 1, la do«-a taat 41 «« iba ayaptona produce« by 2,4 -0 varia« fraa a Ñ« a a ilJ ataxia ano atlfTaaaa lo tba b la l '..«a to a « a fta ita ayoteala. IfeltlaX c ffc c ta aa.-a o ftaa nota« b Ouura a fta r tba oral a b a la la tra tlo * of 2.4*6. .'tía aataala aar* «cura Tulat taaa a o r v l aa« a altgfet atab la aaa oaaaalnfeal ly preaos: at th la t la a . Iba blad l ^ a «ara alaoya affaotad f l r o t , tba faralaes la ta r » rhara aaa uaually a p r o n ta s Iva lfearaaaa la apa aa la aot at a l l . tba oui« la .a aaa je lata« attb laoraaalm atabla aaa ama «asa apealaos anal? aitaooa« tba hind lim a la a ip aatla aoaamat la a tla « a faa aoaefeás. Tfea ■nao j-rk aaa atib ar n o r » l » hyparactiva. la toa «arly a ta .a a * t u ia t . aaata pélaosla« tfea afeifefelfe otra aamll? If tba a a im l la llf t a « aa« M a to aalk tba «ffesfe aa« a tó a la ara b o tica « , «at «feas aalk*« t - ;v " ■ ■•••Vf •' ••••:• . "ilciiftittti^ t f f H M hjr 2.4-0 tt«i ik* poMiki* m t r U a 0.7- -** T m ••¿yâCs» ; . • tai H * ' t u * ■ - - - ' l* , & ¡ 'r «og « w i t t u t a t r a K h * par ««at of t r f a i toa A ■a» M t praaaatt U dog Ma. 104 ja « t h f a t 4 a • i ' ' ■; ".•*•2 a > ) a n | n i a > ta Ma ft**- • lie b t dlfffeaa rodaoaa o f ta* daadaaol a n o —. b. r C rr»a — U n t o . n *ro ta * t a y r o i d . a d r e n a l. a * a r t , o l a y w a t p e r io d . *oa a o a ls a lfla a a t a ra r o r k id a t r ta Utk* aalMla that a r r l r t t i « t iaor**ao l a aoa* a t t a * oc**» « a le t ta «aa bo w w 4 in t-v» •■'U, H and n a tn*t r * e » i r * d ta* h lA o o t doro o f 2 ,1 -fi aad A o dlad tu.-lrv u>* « ip -r ta » o t. C iio m it.a asi g ^ C J • ■ 1 U " a !»OU U -i u«*»-pT»«*a V J : «V *0 4 p C U rh* a w t o a o produced by ta* k l^ i a la c i# oralo doaoa 0 -a itw y «aa not o o * * r r * d t a t a * p r o a a a l atudy. i i* rr r.- » « . jy ia . . i' tu« acut» iiM a *n o **d * a l l d - u ; -.«<» * imi * d c a * i .t n i i l l y » r * * * r--* S i . t i - r du**.» io * * . a *« tn f a i a ^ l* *.n» i » *t»o tr.» a r l a 'f i r r i t a t i o n . end u.* in n a r i ir. a , *r.«o l i f t * « ta . *pp**rod to b* boaorrbacla* ..raaucad « n a r a t t a a ad «oaaoqaoat N l ^ t la y a * f t * r Jw i .a u t « : o f A a dosa d lffu aa r*da*aa y point to • (p in o l w oaafer*! l aal A» M* tarant* atady ta* «ara aure lead doaad f ! • 5 **d 19 A f . (« -ô per t i l o o f ( i d ; o a t^ tt. Ocaoa o f cO « ■ par fella . k M W t 4M a «r " t ;ñ 5147 produca daatb aban a d a lb is ta ra d o**r « ¡a r la d a t 11 to «•} a o j o , a o a fa -m lty « l u rb ia la la aub-aauta atu d laa la Ic a a ¡a r f-r a a d by M ill aad l a r l l a l a ( i l s rfeay a d a la la ta ra d 2 ,à -0 lntraaan ou aly la dooaa o f 25 -T j l a<. par d i o f » b la j a e t l o n * . oueb « n ia a la d a d 2 0 >72 ü m t 17,SoO yt~ u tt i m a 2 e » À_____ 1_____ B L W 13.82 5.33 1 5 .* » 83 23 13.70 4.08. H.3V> 71 27 <4i 0 4.01 10.800 11 12 36 14.72 " 4.44 13U5» 73 21 ...Hatt,., 4.14 - Z * l> 9 - - 2 3 — 5.12 >47 13.550 •J J2 8.74 > 8 8 I 8 0 OO 12 Î7 4 .« « -U . _2L 41/ , . 2 . V - 1>31 > 8 4 14.450 54 33 23 54 :■ 36 14.81 > 3 8 17.850 ; , v 30 14.74 4.97 15.530 70 30 ¿ S S T a r r y ™ < r 10.43 4.55 20,450 20 71 47 “ 30 12.47 4.47 14.200 ?3 ^ -. ‘S.\ ' ___ 22.-19*28-. 3.42 -1 3 .1 3 2 -—SL—— 12— 0 14.06 5*92 14.350 22 43 42 ¿5 36 14.53 5.74 * 15.55« 11.57 10 o 3» h * tf« 4 9 9 “nr -M ----- 14*34---- 7.03 0 1404 * .3 3 30 14.78 4.21 11.4S0 15.200 17.500 TT" 30 0 30 17.800 17.800 14.40 13.35 405 4.24 M M U.;« uTT 11.1 U 3#* * 0 *20 18 73 60 15 28 71 -ÏL- 34 45 36 10.43 12.44 99 11*32 V. ft 4.02 4.14 0 13m > 36, 14.44 5*86 > 22 22 545* 15.400 *4 U.850 K U | Im o 3 3 3 0 2 3 2 0 9 5 5 3 . ' 0 • ' 7 *. 7 5 * 0 .0 0 « •* JN Í «a* <* 19 0 3 litt t lT lM 3 2 54 -44. • to 28 3 1 14.34 i¿i ^ 4 7 m13.15» * • a — 1 12*199___ 12____ 22. 44 a - i Ak *t ¿uT 0 0 13.3 . 8 0 0 4 * 2 7 5 ¿LÍL. .. 0 0 ■o 3 II422 ___ p ______ U ____ L 47 73 . 3 3 / 4 Jr ^ 4 l 15.000 77 X . .1 .7 .7*31_U a i S __ 14_______ i____ L. 3 9 - 12»33___2» •20 17,350 17.300 30 • o 5 3# 30 » i&iM 3*191 -U - H I H a u iiU r H la 4 7 -ii. 11 JL. _L t la * .4 . u 4 » la c .a , ft TLÆt.ff‘i'V’7 M. 5155 Made in United State» of America ARNOLD J. LEHMAN Chief, Divirion of Pharmacology, U. S . Food and Drug AdminUtration MNO02 527 CHEMICALS IN FOODS: A REPO R T TO T H E ASSOCIATION OF FOOD AND DRUG OFFICIALS ON CU RR EN T DEVELOPM ENTS. PA R T II. PESTIC ID ES SECTION II. DERM AL TO X ICITY Pesticides may have uses other than those related to agriculture. A num ber of them may ñnd clinical application in the control of ectoparasites on m an and animals, some may find use as insect repellents, and still others may be useful in treating fungus diseases in human and veterinary medicine. The development of these various uses involves application to the skin; consequently, dermal toxicity studies are of considerable impor­ tance. M any of the pesticides are not amenable to clinical use, hence, the list of compounds investigated for dermal toxicity is not as extensive as th a t presented in Section I dealing with acute toxicity. The general method of approach in evaluating dermal toxicity was to determine the effect of a single acute dermal exposure, the pesticide being employed in the dry form or in solution in an innocuous solvent, dimethyl phthalate. If these initial tests indicated a favorable toxicity, subacute studies were undertaken. These were either daily applications of a series of dosage levels for a period of 21 or 90 days, or occasionally wear-test experiments in which a definite quantity of the compound under test was incorporated in each square foot of cloth and the animal permitted to wear the treated cloth as a jacket for 21 days. Only limited wear-test data are included j n the tabular summaries. Frequently animals failed to survive the scheduled 21- or 90-day test period. This is shown in Table I I I by indicating the number of doses which were fatal to the animals. Dosage levels were determined to a great extent by the proposed use o f the compound. If the results of single or multiple exposure experiments indicated th a t the compound was too toxic to warrantfurther consideration, additional experiments were not done. This accounts for the numerous instances of listing dermal toxicity values as less than ( < ) the value pre­ sented in the tables. Identity of the compounds listed in Tables II and I I I will be found in the preceding article of this series.1 In summarising the results it may be stated th a t insecticidal materials which are solids or formulated as wettable powders and dusts are poorly absorbed through the skin and a single massive exposure may not be serious. A t least, if prompt remedial procedures are invoked immediately after large quantities are spilled on the skin the chances of injury appear 1 Quarterly Bull., Assn, of F. and D . Officials of U. S ., XV, N o. 4, p. 122. 3 DOW 882090 Reprinted from A w o c u n o x o r F ood Ac Dptra O m c u u o r t n U m ttsb S t a t u V oL X V I , No. 1, JnnuAry H U t GGG5G10 4 ASSOCIATION OF FOOD AND DRUG OFFICIALS TABLE II Dermal toxicity: Single acute (24 how) exposure in rabbit* vxsn cD i am oxncan LDUMO./EO. T O U nilA T lO X sm u Lxm cn H B i a t u r im Plant products and derivatives Botenone 10% technical end» in dim ethyl phthalato >•«0 Pyrethrin» 20% pyrethrina in aoya b*»a oil >1880 Allathrin Nicotin» 8 w u U » in Undiluted alkaloid 80 Ryania la powdered to m >«00 Slight body anight to e, generalised wea Irate», partial paralysis and iaocftrdination of aetremitiea No d p iillim t local No systotaie affacte •Ida effecta; severe irritation of abraded »kia Severe skin irritation No local »kin effects No effect Convulsions, respiratory failure and daath No »yatemic effects Thiocyanates Thanite (.0 ml. Lethana-60 Undiluted technin»! (rada 60% coaeanttate in petroleum distillate Lethaoe-3St 50% eoaeentrate in pe­ 0AS-0J mL troleum diatillate Lethase Special Lethane-60, 3 parte L»thaaa-38t, 1 part lO Jm L L0 ml. L ittle evidence of pononiag until lethal dcae 1»reached Ifodarate »Ida lrrita- Little evidence of p»»— «'m tioa due to petro­ until lethal data ia ap­ leum diatillate proached No significant local ir- Extremely toxic; no «am ­ ritatioa ine signs of poisoning until lethal doaa ¡a ap­ proached. For em plom e eee Table I Slightly irritating due Aa for Lethane-60 sad to patrslaum die- Lethana4M tillate No local »Ida affacte Chlorinated hydrocarbons DDT DDT Hicrotiled powder *0% wettabla powder D ry technical grade 30% technical grade in } Apparently not absorbed, hence no evidence of toxioity No local akin irritation Slight trem ors, anorexia, and emaciation. Complete recovery in 8 to S days Köderst» akin irrita­ Hyperexcitability and ecotion vulaiona prior to death Fatal outcome delayed 7 days poet exposure Moderate »Ida irrita­ Symptoms of poisoning but tion no deaths; raoofscy in 8 to 6 days Blight akin irritation No vysBptoma of task ity « Koderate skin irrita­ HypenxcitebUlty Indicat­ tion especially of ing soma absorption abraded area» No effect >3830 J im a th y i p h th a la ta TD E 30% technical grade in dim ethyl phthalato 1300 TD E D ry technical grade > «00 Methoxyehlor TBB 30% technical grade in dim ethyl phthalato 30% ia dimethyl phthalato >3830 >1880 COOSGli 5 TABLE II —Continued t n S T IO B t V O B M D lA H O S r i AmOXXM ATS LDmK O . / K O . o i u c A i x m c n tm m c tm c n Chlorinated hydrocarbons—Continued UsdftM <150 2000 No aida irritation dimethyl <7*0 No aida irritation teoh&ical >4000 teehnscnl >4000 No orideaca oi aida ir- No erideneo a t poae eyeritatioa tsmio effects Ai lor prolea Ax {or prolan >4000 dimethyl >ut 1% in vanishing eraam baas D ry (n q rJ form ¡0 2% Toxaphene Moderate sida irrita- Seven rymptoma, oobtuInone, ro o m y ia 6 to 1 tioa days No aigntdeant local ir­ I H yperexdtability proritation ; peering into eonenWoaa No significant local ir- 1 end death ritetioa Moderate aida irrita- H yperexdtability, b a t d o deathe; reoorery in 3 to 3 tioa day« Moderate liria irrita- Tremora Iaadiac to oontioa Tuleioai aad death Seram «Ida irritation Tremora, eoaeolaioai aad death Serem tetania type of eonNo akin irritation ▼uliiona aad death No aida irritation L ikaaldria D ry form ta >4000 <710 Heptachlor 20% ia dimethyl phthalata dimethyl 20% ia P^thaiaU dimethyl 4% ia phthalete dimethyl 4% ia phthalete D ry powder Prolan 20% ia phthalete Undiluted Undiluted grade Chlordaae Aldrin Dialdria Bulas Stada 1400 <300 S lith t edema of akin Ataxia aad muaeolar weaknaaa prior to death Mild symptoms as sbovs 81i|ht edema of akin but ao deaths Moderate akin irrita­ Secern depresión, m aeeaier tion cepecially of paraiysid and death abraded areaa C005G I2 5158 DOW 882092 ASSOCIATION OP FOOD AND DRUG OFFICIALS 6 ASSOCIATION OF FOOD AND DECO OFFICIALS TABLE ll-Concluded AmOKDCATI raucvumox tursexDE LDu no./»». rrttnezexffscxi DZUULimen Herbicides—Continued 100 1%in n t v E^dothsl Hálete bydr»- Technical grade powder Iqrb aid* in >4000 1 Severo aidn irritatioaj Afiortn* ta d hwnoturie specially oi abraded beten death am a No local alca offacta No eyntemle affocta Activators a-Propyl 4% in boma phthalata Piperonyl 30% in butoxido phthalata Yaa Dyka 2M 8% in phthalata Sulfoxida U ndiluted (rad* Fiptroayl cy- Se» tabla m elohexaaona dimethyl >37* dimethyl >1880 dimethyl >470 technical > (.0 ml. No akin irritation Ko •yvtemio eflecte aotod a t thia doaaca Hyperexcitability and oonNo akin irritation ▼nlaioaa but ao dootho SUsht irritation of aidn' S ich t anorexia and «nicht kn Moderate akin irrita- No eyatemio effeeta taon TABLE I I I Dermal toxicity: Multiple (repeated daily) expoture in rabbit* yssnass Am oznun LDm, MOe/KO. HUltV« Plant products and derivatives Rotenone 100-200 Pyrethrins 200-400 Allethrin Nicotine Dihydrdrotenone 40 100-200 No deaths at 100 mg. during a 21-day daily ex­ posure period. No survivors after 12 to 14 doses a t 200 mg. No deaths at 200 mg. during a 21-day daily expoaure period. No survivors after 8 to 14 doses a t 400 mg. No injury up to 1 gm. per square foot of cloth during a 2 1 -day wear test Based on a 2 1 -dav test. In a 21-day wear test no survivors after one exposure at 2 gfh. per square foot of cloth; no deaths a t the 1 gm. per square foot level No deaths a t 100 mg. during a 21-day daily ex­ posure period. No survivors after 10 doses at 200 mg. 0q05G I3 5159 ASSOCIATION OF FOOD AND DRUG OFFICIALS 7 o TABLE III—Continued nSTtCZBX AVPSOXOUTS LDit, MO./KO. levisr* Thiocyanates Thanite < 1.0 ml. Lethane-60 < 1.0 ml. Lethane-3S4 Lethane-3S4 Spe­ cial 0 .1 ml. 0.5 ml. No survivors after 4 to 5 doses a t this level; effects of lower doses not determined No survivors after 5 doses at this level; effects of lower doses not determined (Approximately half of the animals died after 8 | doses during a 2 1 -day daily exposure period Chlorinated hydrocarbons DDT 150 TDE 200-400 Methoxychlor 600 TBH < 200 Lindane 20-50 Toxapbene Chlordane 40 20-40 Aldrin <5 Dieldrin <5 Heptachlor <20 Prolan 400-800 Bulan <200 Parathion 5-15 - ■ ,L__________ Based on a 90-day exposure period. Intercurrent infections resulting from lowered resistance brought about by the chronic posioning effects of DDT appeared to be contributing cause of death. No survivors a t 300 mg. after 10 to 14 doses Severe symptoms but no deaths a t 200 mg. during a 90-day exposure period. No survivors after 6 doses at 400 mg. Based on a 90-day exposure period. At 1200 mg. all animals died after 7 to 19 doses No survivors after 5 to 8 doses. Effects of lower doses not determined Severe symptoms but no deaths a t 20 mg. during a 21-day exposure period. No survivors after 2 to 9 doses a t 50 mg. Based on a 90-day exposure period At 20 mg. one animal died after 51 doses during a 90-day exposure period. No survivors a t 40 mg. after 10 to 26 doses No survivors after 4 to 10 doses. Effects of lower doses not determined No survivors after 14 doses. Effect of lower doses not determined No survivors after 14 doses. Effect of lower doses not determined Symptoms but no fatalities at 400 mg. during a 21-day exposure period. No survivors a t 800 mg. after 6 doses No survivors after 3 to 3 doses. Effects of lower doses not determined , Severe symptoms at 5 mg. during a 21-day exposure period. No survivors at 15 mg. after 3 to 5 doses < oo 00 ro o CO 8 ASSOCIATION OF FOOD AND DRUG OFFICIALS TABLE I II —Continued kstxod* Amozoun ID«, HO/K& ip fu n Organic phosphates Dimethyl parathion 25-100 Paraoxone <1 EPN T E PP <5 ASP-47 1-2 E-838 10-40 Symptoms of poisoning but no deaths at 25 mg. during a 21-day exposure period. No survivors after 2 No survivors after 14 to 15 doses. Effects of lower doses not determined D eath of one animal in the group exposed a t the rate of 1 gm. per square foot of cloth towards the end of a 2 1 -day wear test No survivors after 9 to 13 doses. Effects of lower doses not determined Severe symptoms but no deaths a t 1 mg. during a 21-day exposure period. No survivors after 3 doses a t 2 mg. Severe poisoning but no deaths after 40 doses at 10 mg. during a 90-day exposure period. No survivors after 11 to 15 doses a t 40 mg. Herbicides 2,4-D Endothal >40 Maleic hydroxide >40 No toxicity noted after 1 m l./kg. of a commercial product containing 15% of the active ingredient applied daily for 10 doses . :i No significant systemic effects a t this dose during a 21-day exposure period. Higher doses were not tried iNo effects at this dose during a 21-day exposure period. Higher doses were not tried Activators n-Propyl ¡some Piperonyl butoxide Von Dyke 264 Piperonyl cyclo­ hexanone Death of one animal a t this dose during a 90-day exposure period. Higher doses were not tried Based on a 90-day test Symptoms but no deaths a t this dose during a 90day exposure period Death of one animal a t 100 mg. after 45 doses during a 90-day exposure period. No survivors^ after 4-10 doses at 200 mg. to be slight. Single massive exposures to insecticides which ore liquid at ordinary temperatures, or solutions of solid materials in kerosene or other petroleum distillates, must be considered as dangerous. C005G15 5161 ASSOCIATION OF FOOD AND DBTJG OFFICIALS 9 882096 Repeated daily skin exposure to insecticides, especially in solution, con­ stitutes a real hazard. As a rule many of the materials in concentrated formulations are skin irritants which serves as a warning of dermal contact. However, in dilute solutions this warning sign is lost and because of the delayed effects of a number of insecticides, serious poisoning m ay occur during the handling and application before the danger of poisoning is realized. G t COOSGiG 16 5163 \8 *v - .-.v .■Ì*':v' & ® Mowhsaloal Reseeroh Deportami '■vi' 12 5084 »ob jM t resulte i ',*>#53 1. -%** THE DOW OBKZCU. COMPAIT or s m ^ zn tm rzoR resta o t m m :g m sjwsE CORREHT PRODOCTIOH 2 .WDICHLOROPRBROmCSTZC acid aì COMPASSO WItH PAST PRODOCTIOH MATERIAL. Pln ' d ;VJ- Work Bfr ■ «•pt. To Check Central K t i u n h Index, Bxeevtlve Reeearch Conlttee Texas Pile A Western Central Reseereh Pile * < ir V - s s - i i. • -- .•_! * «a* H. 1. Borie r - - . . ./ f-f V |T Brlttaa's Divisimi Attni R. C. Dosser C. A. Hlghblll ¿¿m m «r^rer o,ep|cér? A P S i -j i s s l ^ Medleal Depertaent Attni H. H. Gay '• 3 Safety Depertaent MaeCnteheon Atta* - ■v * A : — «»* IhLrh L.G.f. APRn i« Aarlonlturai Chealeals Divisimi, 1^*7 Bidè. Atta« I. V. Brlttoa W f l M The skla Irritatine propertles ©f enrreat prosastica:;.' 2,V-D «old ere stallar to those of 2,WD eold preparo! hy tha\ aev aethod latroduoed in 1950. 2 vV-Dlohloropheaax7aoetlo aold Fheaoxyeeetlo soldi 2(Wlehloro», Aoetle soldi 2,Wlshloropheaoxr*t Aeetlo soldi phsnaxy-, 2 #'*-dlehloro-, 0( TMIS KEPOST » THKPftOPCKtY. . ’O f . ' v ' v « ^ « TNI MW CRtM.tU CdHPAHT . -rii 515 4 / rt >V psUV a ' J gw g* *..■ rT 23 .1V - U . * 1 " ••’ , ^ 'm * « • 2 m ■ ■- '" r .v à s S c â ÿ * / rf}»‘ ** A f t v i u « i o f dermatitis baro . ooanootlon w ith tho a a a u fa e tu re o f 2 ,e - 0 o e ld by 9 » Bov I jp-.'-iJ: • Company, •••• A s a a p le o f c u r r a n t p ro d u c tio n a a t s r l a l . - f o r s k in i r r i t a t i o n to o ts f o r com parison w ith p a s t p ro d u c tio n ■ s to ria i, SfflgMBMS' 1, ' '> ? Tbs s k in i r r i t a t i n e p r o p o r tla s o f « o rro n t 2.W-D s o ld aro s i a i l a r to th o se o f 2 V aoiA p roparod b j r t h a ao th o d in tro d o o ed i n 2, 1950, -V Oaap 2 , 14-d io h le ro p h e n o sy u o e tio a o id i s m o d e ra te ly ‘ i r r i t a t i n e and s tro n c s o lu tio n s o f th o a o ld a ro s l i ç h t l y t o th e skin* on proloncod and .re p e a te d s k in o o o ta o t, 3, irritntind ■v S?**** • HI - • o ifth a , D e rm a titis w i l l p ro b a b ly r e s u l t i n some su b ì s e t a a to r i a l , p a r t i c u l a r l y i f w e t, i s allow ed t o e o n t a o t t h o ,1R s k in re p e a te d ly o r f o r proloncod p e r io d s , ._>*■«*;t s i ^ œ ï î • V . ' f •/• MATERIAL Haaet 2,W"»Dlohlorophanoxyaeetle a o id ÿ-: Form ulât Cl S tr u c t u r a l ! Cl ■ S a p irlo a li CqH^Oj CI j B ,R ,0 . H i e H o.! B ,R ,0 , X H o,I Bour s e ! o H 0 i. . 2 • . . 0 - .C - C - Œ ' ’ - T 23.1V -U -9 2372-21 P la n t fr o d u o tlo a Saap io o b ta in e d by! B , R, Hoyle ;'-«VVi v ■0001-7 ••• ** 4 jr% h. ' t « • '^ e " rf-H* J p f T 33.1V -11-9 pm * 3 SS©$,':7r . rasramra uma +* When d ry 2 , h- 0 mold w i bandaged re p e a te d ly o n to then «90?.^":* ■V■/-*<,*'•"' shaven abdaeon o f a r a b b i t , f i r * days p a r w t k , f a r a t o t a l Cf;v 30 a p p lic a tio n s l a 1W day*, v a ry a l i g h t s k in i r r i t a t i o n o f guest ~ * ¥j0'$i-;r ■' a b l* s ig n ifie s * » * r e s u l t e d . •# 4<^ % % kben daap 2.W-0 a a ld aa a ta a ta d l a sa a a n a a a a r , a o d o ra t* «Ida I r r i t a t i o n o b a ra e ta rls a d by hypo r aa l a p i * & ' ;4S : fcSCVs*'- aa d a n p a r f l e l a l n a o ro a la r e s u l t e d . ' Repeated a p p lle a tlo n a o f a lOjf aaapanaloa o f th e a a to ria l;< l a « « to r to th a o a r and shaven abdoaan o f a r a b b it prodoaod a l i g h t * ; w . a k in - I r r i t a t i o n . ■t>r Repeated a p p lle a tlo n a ' o f a l.O jf aaaponaloai o f 2 ,W > a e ld l a a a to r t o tha a a r and shavan abdi a o f a r a b b it •■iv j'C'V -:^*A prodooad v a ry a l i g h t a k in I r r i t a t i o n on th a abdoaan. •r* Caaaa o f d a r a a t l t l a have boon obaerred l a e e o n a e tlo a w ith ' th o a a a n fa c tu r* o f 2 tV ^ ie h lo ro p h e a o x y a e * tle a a ld l a th e p a s t o a a s t a t e r o f o o o aalo n a. Tha p ra a a n t p ro b la a than l a r e a l i t y l a an o ld p r o b la a . ■■- ■«♦ *> - A rev iew o f tho eo n e la a lo a a roaehad l a to a& eo lo g lo al t a ta d la a oa 2.V-D a e ld re p o rte d by V, K. Rove oa 1O -12-J0 (B lo o h aale al R aaaarch Departm ent P ll a R oa, T23.lW-U«3 and •'j/iA'ifcj • '‘i t M t 2 3 « l h * U ^ ) f o llo v a i 1 . " I t would sa a a fr o a th e se r o s a l t a t h a t ' la p u ro 2,U«4> l a nor* l i k e l y to produce a s l f n l f l o a a t aa o u a t o f a k l a ‘ I r r i t a t i o n th a n la pur* a a t e r l a l i aad a l a o , I t aaaaa t h a t a daap p ro d u c t l a nor* l i k e l y to produce I r r i t a t i o n th a n l a a d ry i i i .•«Sag 2 . "Although n e i t h e r th o a a t e r l a l p re p are d by th o o ld o r th o now • e th o d l a a s k in s o n a l t l s e r , th e d if f e r e n c e In a b i l i t y to praduoo.1p r la a r y sk in i r r i t a t i o n l a s u b s t a n t i a l and th o lik e lih o o d Of i k l a 5* 1 i? !? !’) i \J ■> o o o i - 7 . $ # a ( - U X ’**. a . 9; J S „V.** - - « - ^-«o® t*- CT' %■ lì. ff t .. ■ í '■ ■ ■ V'tV-'"- 5 1 Z*J >. • * ,r : # ^ ^ p ï w à ^ 3 M l e r : A •:ife^#+jitoaterniCentral Research File tttcnTs-'iDivision Agricultural-Chemicals Division Attn: J. W. Britton : \ -i- ‘ "i ’ ' ■■•,.. s * i » « h . 1-1* . 7 Bldg. TTM*1 Field Agricultural Chemicals Research, v ¿ ’ $Attn» W. C. Dutton ..-’ ^Attn* . .■; , , •:;--2i'r_r¿.Vä '.';,V • *•• •*••• -■ r.' • v -:í .* ^¿¿Agricultural Chemicals Laboratory,:Seal’Beach -í/^'^Attni ' J. F. Kagy • V i. ^ ■'•'V-¿V •. :' V v¿'.' ’Vv>£ Agricultural Chemicals Research Laboratory Attn* G. E. Lynn J. S. Johnson Medical Department Attn: JJ. H. Gay Safety Department Attn: S. M. MacCutcheon UNIT INDEX Results of range finding toxicological tests on • 2,V-dlchlorophenoxyacetlc acid prepared by a-new method are , presented. Health hazards and precautions for safe handling are given* TOS REPORT IS THE PROPERS OF THE DOW CHEMICAL COMPLY C0017C4 5169 j •*. *;•£<■*’ B 3 ^ w i - h *«• * ;, * ■ - •:• :.** * i & . - ■ '-'i J*'4M dBfc ' ■*•"£l**• 5n?tf*'c4 A cetic a c id «(2 ,V-<1tohTnrophouoxi)-, 1•■"5.4 - v -■ • IT. ■ ■ ■ * .— ■ ■ ■ s*H& . — „ 11 . . . 'fii A aew n e tte d fo r p rep arin g 2, t ^ !l o h ioru phana«yi< ¿/-f.:;■««J «old m s ln tro d eed In 1950 which g ars a w h ite r and o d o rlaaa p ro d o e t. What a re th e ta x ic o lo g le a l s u b je c t n o ta r ia l p rep ared by th e new aethod o f hew do th ey eoapare v ith th o se o f th e o ld e r p re d n o tt. -^ _ soRarera? 1. :7 • # • is *s v -1 y f ’& iift'* o ra l to z lo ity . 2. «*.•* •*• -tJ 2,U -D ichlorophenoxyaeetle s o ld l a a o d e ra te i n < The fre e a c id and I t s stro n g s o lu tio n s a r e 'i ^ e ‘•m • I r r i ta t in g to th e e y e s. is*. to th e e y e s. 3. Weak so ln tlo n s a re s lig h tly i r r l t a t v ■:,*%*** . The d ry a a t s r l a l does n o t pr odnoe s ig n if ic a n t a k ln j I r r i t a t i o n on re p eated a p p lic a tio n to r a b b it a k in , b a t a t i . "';y s o ln tlo n s prodnoe s lig h t sk in I r r ita tio n « U. 2 (W D iohlorophaao^raoetlo s o ld I s n o t abosedi * th e s k in In any a p p re c ia b le q u a n tity . V **«• : i¿1?'* ,l * * - V v 5. 2.Woiehlorephenosyaoetls said prepared by the aethod la slgnlfleantly loss irritating to rabbit «Ida than * i oldtr produet. Tests oarrlsd oat oa hnoans a 14 U Howe oa 10-12.J0 (lloohcaleal losearah «o. T23.lW.U-d) hare aeafimed tho oanelaslana laadhol tren* stadias oa rabbits. Together these tests bare shtea that did . . • * !i.» • •. • V . 5170 00017C5 ■^UA^/.L-ÍH II i»><: HÌ •.'.•ïi'A\i >••-i sI ítfS** ■M ■tí- L:?-> í r U V : !í “ ■*( *A‘í^; W % $ •*>1 r - M M U >C rj f u ¡j R *'•?• li * 1 I«» s í» »i ÎÇ*V Kíl&íV Bfì i i! |T .it h M « * ß a ií * ¿ |l ,\H *4) ;t <$ Sí - : . : O • " ,, « • y F £ A '£ !' (í U .4-ii-wríí í ‘ét*#\ ■ i j M I RvF;;: * plíi| C Ì !í ti 6 i* 'ií» " :i. í ' !>■ i i fr­ itó' B: 7j>r i W r r n .,:' :-.r- - ' : ■ an ftrtfrttfli - - 2 ,W le h la ro p h a n o x y a a a tla M id ? v ^ : # - M g g r v t 2A«tirfn Xb ‘ »V v o f th a M id l a propylana c ly e o l m i p laaad 1b th a aya a f o • - ■■ •- • m r t « j« I r r i t a t i o n o h a ra a ta rlsa d by aarkad p a la v Eft ... and a o rn a a l d a a tf* ra a n lta d . ■ Khan a 1 «0J( a o ls tla B o f t h a a a t ‘ ln propylana (ly e o l w u ta a ta d ln a a l a l l a r a a a n a rt a l l f h t ^ tra a a la n t aya I r r i t a t i o n ra s o lta d « •f-i. ¿jfiÜgi la n a d la ta aaaM ng o f f aya v ith v a ta r radnead th a aao o n t o f aya w g » y i w - w « « tln I r r i ta t io n Khan th a d ry 2 ,V-d m IA «u baadacad ra p M ta rtlj sh aran abdonan o f a r a b b lt, f l r a daya p a r v aa k , f b r a t o t a l a p p lle a tlo n a ln 1U d ay a , no a lc & lfla a n t a k la I r r i t a t i o n I B apaatad a p p lle a tlo n a o f a 10£ a o ln tlo n o f th a a n to r la l l n e a r b lto l a e a ta ta to th a a a r and aharen abdi -■-M i o f a r ü b lt prodnead alicht akln Irritation oharMtarlsad by allcht a e a lln a a a and h a lr lo a a . SttU&i&nslaa Zn th a ak ln I r r i t a t i o n ta a ta eontfaatad, th a ra n a ' a a Vt! arld a n a a th a t 2 ,V -d leh lo rep h a n o iy aaa tle M id n a al th a ak ln ln any a p p re e la b la d o a s tlty . * "tmT™ 2 ,V -0 leh lo ro p h aao x y aaatla M id and I t a s t l'-5\ a re a a ra ra ly l r r i t a t l a g to th a ay a« . Tha d ry fr a a M id a p p a ra n tly la n o t tr r lf e a tla c to,. oiM.' ln tM t a k ln t b m r a r , p ra rlo o a m k an th la 10-22-50 by T . K. Bona (S la a h a n la a l Raaaarah I o , T23. 1W U - 3) h a t ahowi th a danp prodaa« t a t a a l l * U y t a th a ak ln an p ro lan cad and ta p a a ta d a a n ta a t, ÜOO ■ ’S* -ff.: 5172 0001797 8 8 » 3 S f o V ;/• ’ T23.1V-11-7 _^>»V 1 ’•■• h|» J «7 th is n o ta r ia l produc e s lig h t sk in i r r i t a t i o n on prolonged J/3?5f ■ta d re p eated ak in e o a ta a t. A ccordingly, th a ra id good re a so n t a 1 ¿Z£?.;r. H av a th a t prolonged and ra p a a ta d sk in e e n ta e t fay hwnans w ith t t r ^ " jp X v r^ daap a a ld o r I t s stro n g s o lu tio n s e ig h t r e s u lt l a th e ^ & o f da* i t l t l s . « li: >9t•*£nu 2,V-01ohlorophenexyaeetle so ld I s a o d a ra te ly to rlo sw allow ed. Although i t would ba p o ssib ls fo r a p arso n to ^ •* # ¿ 3 '* ,; a s u f f lè ia n t anouat o f th a a a ta r la l to ba dangerous to U f a , th a ra ; • ■'^ *i I s s lig h t h asard from in g e stio n In o rd in ary In d u s tria l o p e ra tio n s , PRECAUTIONS TOR SATE HANDLING ■ 't-fi Z t I s stro n g ly recesneaded th a t ays p ro ta a tlo n b a re -- . .¡b**SVs - q u lre d ah a n ara r th a fra a a c id o r I t s stro n g so lu tio n s a ra h an d led , -Pace s h ie ld s , gogglss w ith sld a s h ie ld s , o r -the e q u iv a le n t «111 v ig il - . a ffo rd s u ita b le p ro te c tio n . S u ita b le f a c i l i t i e s fo r w ashing th a 4r ay es w ith w ater should ba re a d ily a v a ila b le . . . ■„ ■ ■ ■ ■ I fre c a u tlo n s should ba tah sn to ' p ra v a n t prolonged o r re p eated sk in c o n ta c t w ith 2.^ -d lch lo ro p h sn o x y aeatlc a c id , : e s p e c ia lly w ith th e dasp a c id o r I t s stro n g s o lu tio n s , such as »•it n ig h t oacur fro a w earing c o n taa ln atad c lo th in g ■ ■ ■ tV- or f t f a ilu r e to wash th a sk in soon a f te r c o n ta c t w ith th a n o ta r ia l, ^ ^ 5’ lib b e r g lo v es nay ba s u ita b le fo r p rev en tin g c o n ta c t cn th a b a h ts * « r is ta . IgAT TO DO POP E2P03DHM 1i - ■ ■ ¿itj I f 2 ,b -4 1 sh lo ro p h en o sy seetle a a ld e e a ta e ts th e e y a s , «.»fd th ey should ba «sahad tharauahir w ith flow ing w ater fo r at least f if te e n a la n te s , and soon a s u o s a lb la . ■-a \ c In th a ev en t o f c o n ta c t upon th e p e rso n , earn ...■‘- v 5173 0001733 i jSÇ/', ? W $ ß ? % W & k S.~ : ••• •‘ •53aJ*-M* ' * • o r ahoaa should to w i w d «ad t t e s k ia th o ro u g h ly « i« b M ap « a l « t o r . U r tili! i t t t n t t l . lt atela i r r i t a t i o n la » « » ra n t o r l f « er p a ia p e r s is ta . C oats * a lo th in g should t e u tsh ed thoroughly b efo re ro -u s a , Xf 2,b ^leh lo ro p b en caq raeetlo «o ld should t e e * l l » * d / » .•'•■■ .■ .*■ »• '•.•"■»i'»*.--^S I tin e sh ould t e Indnood p ro a p tly by tin k lin g S to th ro a t vittoJKT r *.•")>. t p'~ / fin g e r o r by c a r ta i 111 a s » soap s o lu tio n o r a a lt s u t ( 2 ta b le sp o o n fo ls o f ta b la s a l t l a a g la s s ot u n ta r)» i l e a l a tta n tlo n th an should te o b tain ed , a t o aaa. V«-»V ' ••> *.*vr Í | v , ? A ,- ■’■S-.i JL’ -’ -V* - - ■W'V, .* 0001739 <2 o % I V I t c f 'r i n t n l (m m A s h n h r M r t o r F» m . h **: !> i:v*u O i i v i a i . k c.7 m i : I ' h i t m » .St a t i c V*.|. ; : v j # No. V.'Si CHJ&HCAIJI I^iOTO: A HKl’OHT TO TIJK AiiriOCIATIOX OF FOOD AX3) DltKO OFFICIALS OX CURRENT DiiVKLOI’i.IiOX'i'3 •vj Vj4 ro 00 P A R T I I . PESTIC ID ES o S E C T I O N III. SIJI* A C U T E A N D C H R O N IC T O X I C I T Y f ARNOLD J. LEHMAN, M.D. C h ie f, D iv isio n o f Pharm acology, U . S . Food and D rug A dm inistration DOW S e c tio n s I an d I I in th is se rie s o f a rtic les on p esticid es d ea lt w ith a c u te a n d d c n n a l to x ic ity , am i a p p e a red in t h e O ctob er 1951 a n d J a n u a ry 1052 issu e s o f t h e lh ille t in , r e sp e c tiv e ly . T h e p resen t a rtic le summ arizes- th e su b a c u te a n d ch ron ic t o x ic ity o f n n u m b er o f im p o r ta n t p esticid e s. R eca u sc o f th e tim e a n d effo rt in v o lv e d in lon g-term stu d ie s, t h e lis t is n o t a s e x ­ te n s iv e a s t h a t fo u n d in t h e first tw o a rticles. T a b le IV su m m arizes th e su b a c u te a n d ch ro n ic to x ic ity in ra ts. I t w ill b e u n d ersto o d from th e w ord ­ in g o f t h e ta b le h e a d in g s th a t b e tw e e n th e lo w est le v e l w ith groas effec ts a n d th e h ig h e st le v e l w ith o u t g ro ss effec ts th ere w ere n o in te rm ed ia te - d o sa g e le v e ls. S o m e sp e c ia l e ffe c ts o f th e ro d en ticid cs in ra ts are p resen ted in T a b le V , a n d T a b le s V I a n d V II su m m arize th e h ig h lig h ts o f ch ron ic stu d ie s o f ce rta in h e a v y m e ta ls a n d ch lo rin a ted hydrocarb on in secticid es in d o g s, flum e c f t h e s e s tu d ie s a re still in progress a n d therefore certa in co n clu sio n s in t h e p resen t p a p er m a y bo altered in th e lig h t o f later d a ta , a n d certa in m issin g figures v/iil e v e n tu a lly b e su p p lied . I d e n tity o f t h e co m p o u n d s w h ich a p p ea r in S ectio n I I I w ill b e fo u n d in t h e first a r tic le o f t ills se rie s.1 S U B A C U T E A N D C U It O X IC T O X IC IT Y IN c c c K RATS T h is is su m m a rized in T a b le IV . A n y s tu d y o f less tiia n a b o u t a y ea r m u st b e classed a s cu b a cu te . A p o in t o f in te r e st in th e ta b u la tio n is th a t th e re­ sp o n se o f a n im a ls t o ch ron ic in g e stio n o f a p esticid e d o es n o t follow a s e t p a tter n . F o r cx a m n lsj in t h e fe e d in g o f rotenon o there is a g o o d p arallelism b etw e en le v e ls fe d a n d d egree o f e ffe c t. T h e le v els o f fee d in g d escen d from a v a lu e w h ich p rod u ces e ffe c ts su c h a s lo ss o f a p p e tite , retard ation of g r o w th a n d in so m e in sta n c e s e v e n sy m p to m s o f p o iso n in g , to a low er v a lu e w h ere n o e ffe c ts a r e ob serv ed , th e n to a n ev e n lo ver v a lu e a t w hich o n ly m icro sco p ic c h a n g es in tissu e s a r e se e n , an d fin ally t o a v a lu e w hich p ro­ d u c e s n o e ffe c ts. T h e r e a re so m e ex c ep tio n s to th e rule th a t th e d ieta ry le v e l, a t w h ic h e ffe c ts a s en u m e ra ted a b o v e are p rod uced , is a lw a y s h ig h er th a n th e le v e l a t w h ich p a th o lo g ic tissu e ch an ges nia!:o th e ir a p p ea ra n ce. 1Ounrterly Hull. Anjn. of F A D 0 :Tk:i::!j of U.f?., XV, No. 4, p. 12J. 47 C C 0 4 S 5 X 5176 TA1JLK IV Subacuto anil Chronic Toxicity in the Rat DIETARY CONC.ZHTIATIOH 10 n l t l T i t UIIUOII PESTICIDI Timeo ( Uvei V/lth Fe^lint In Lowest Gross Kflccts Vietiti Highest Uvei Without Ctrr.3 Etlccu Lorrrit Uvei With Tiesuc Dusse* Highest Uvei Without Tissue Dsouz* 5 COCO 2 1000 Natural Products und Derivatives H otenono.................................... Pyrethrins.................................. A llcthrin..................................... R yania......................................... Dihydrorotenono...................... 00 0000 10,000 15,000 100 104 104 10 10 10 25 .1000 eooo 15,000 10,000 25 O 101 101 2500 40 1000 10 500 10 250 2.5 101 104 101 104 101 101 40 3* 10* 45 1000 10,000 10 0 .5 3* 10* 15* 500* 10,000 0.1 15* 600* 2COO 2000 Chlorinated Hydrocarbons D D T ............................................ T D E ............................................. D F D D T ...................................... M cthoxychior............................ T b l l ............................................. A BU ............................................. BB IÍ............................................. D B II............................................ Lindano....................................... Toxaphene.................................. Cblordanc.................................. Aldrin.......................................... D icldrin...................................... Prolan.......................................... Bulan........................................... 100 400 1000 200 100 100 10* SCO* 100 400 75 50 50 2C0* 1000 104 101 23 101 101 101 101 101 104 101 104 10 10 40 40 50 100* 200* 100 50 50 50 100 25 25* 25* 200* 5 ICO* 200* 500 50 £0 10* SOO* ICO 100 2.5* 25* 25* 200* 200* 1 2C0 10 10 50 25 Organic Phosphates Parathi^a.................................... ISPN............................................. ASP-47......................................... H E T P .......................................... Tricthyi phosphate................. D iethyl phosphate.................. 101 52 52 12 31 20 25 1G0 10 130 GO 1000 10,000 GÍW0 1» ) 100 GO 180 5000* 5000 43 CGO 75S576 * TLo Metals Copper (as chlorido).............. Mercuric (as a cc ta to )............. Mercuric (as phenyl morcurie a ce ta te )........................ Selenium (from grain)............ Selenium (as sclcnido)............ Cadmium (as chloride).......... Antimony (os trioxide).......... Barium (as ehlorido).............. © .40 ASSOCIATION 'JO f FOOD AND J3RUQ OFFICIALS TABLE IV—Continued nsncoE T ot« of Level Will, Fecrtin* in Lswnt (jioij £Jscli YVc:ls llfchttt Ixvr.i Viilbnt (•mi Eflccts Lcrcit Level W.:l, ri5,v* Desuse lliclcu Level V.'i.bo-Jl Tit»:i Decer: Dinitro Compounds DN-111.................................... 1000 35 1000 2C0 Herbicides 2,4-D....................................... 2^4,5 -T ........................................ Mcthyi-1-naphthalene acetic acid...................................... Endothal............. .................. Maleic hydroxide................... SO 30 5000 cooo I0G0 1CC0 1000* 1000* 101 17 17 20,000 5C0* 50,000 2500 20,000 £000 50,000 10,000 / 2500 2500 10,000 Activators n-Propyl-isom o.......................... Pipcrouyl butoxide.................. V an D yke 204........................... 17 17 17 ¿000* 5C00* 5000* 6000* 5GC0* COCO* Rodenticides 10S0............................................... A N T U .......................................... W arfarin...................................... Zinc Phosphide......................... 104 104 1 to 2 5 25 100 6.25 (death)* 10 £0 5* 50* 100* Miscellaneous A rasan......................................... Aram itc....................................... P h ygon ........................................ 341 C ............................................ 65 32 52 26 1000 5000 2500 10.0GC 300 1COO 1000 5CG0* 1000 200* 5000* 200 * Lowest level fed. Para'Jiion is an example of this. I t m ay be noted th a t no histopathological changes could be dem onstrated in animals fed a diet containing 100 ppm. parathion for 2 years. This concentration is 4 times th a t which produced sym ptom s of poisoning, and 10 times the level a t which no such effects were noted. O ther compounds showing similar responses are rr.ethoxychlor and E ndothal. A nother point of interest is th a t the LD-0 of a pesticide is of very little if a n y significance for predicting it3 chronic effects. B B II m ay be cited as a n example. In thi3 instance the LDW of B B II ¡3 approximately GOOO 0 G 0 4 G 5 3 5178 ¿¿QSGmmoO OICTASYCONCXU1ATIUII CCMS» » 1 IUi.*.1.1 50 ASSOCIATION OF FOOD AND DRUG OiT7CIAL,l mg /k g , a relatively low order of acute toxicity. However, the chronic in­ gestion of 13BH leads to gross and microscopic evidence of intoxication a t the low level of 10 ppm. in tho diet, a Irish order of chronic toxicity. A th ird point is th a t tho behavior and general appearance of an animal chronically ingesting a pesticide is a crude endpoint us a measure* of chronic toxicity. This is dem onstrated w ith D D T. A dietary concentration of ICO ppm. is ncccssaiy before evidence of poisoning is' observed, y e t microscopic evidence of liver damage is seen a t 5 ppm. The importance of hisiopathological examination of tissue of cnimals on long-term feeding studies cannot be overemphasized. T O X ICITY O F ROD EN TICID E3 IN BA IT FORM | Table V presents a sum m ary of the influence of food on the effectiveness of rodcnticides. These pesticides are usually employed in the form of baited | TABLE V Toxicity of Rodcnticidta LSI, Cf UO./KC. «cowman* Is Bait FciTB A* tbe C hcninl 10S0 Castrix ANTU Strychnine 1.7 1.7 0 10.2 Thallium sulfate Zin. phosphide R ea squill 23 450 3C0 5 10 30 25 (females) 15S (males) 25 450 100-500 (females) 6CQ-SCC0 (males) food, hence, for tho purpose of this experiment various dosage levels of each rodcnticide were incorporated into 4 grams of laboratory diet and these 4-gram portions were offered to starved rats. Animals n ot prom ptly con­ suming tho entire am ount were rejected. Tho eitect of the bait on the oral L D yissum m arised below. A comparison of the values shown th a t the bait reduced tho rodenticidnl effect ot ail compounds except thallium sulfate and zinc phosphide. .War­ farin was not tested in this m anner because its effect is dependent on a con­ tinuous and prolonged insuit to the coagulating mechanism of the blood and therefore not adaptable for study under tae above conditions. S U B A C U T E T O X IC IT Y O F SO M E H E A V Y M E T A L S IN DOGS The compounds listed in Table VI were fed to the animals 7 days a week, the prescribed dosage being mixed with a :1-naphthalene acetic Growth retardation acid c Questionable slight hypoplasia of bone marrow. None of the characteristic hepatic cell changes seen with the chlorinated insectieidca Negative. None of the characteristic heputio cell changes scon with the chlorinated insecticides Examination not complote but it apjiears th at tlioro will bo only _ nl"0,cl," ll'a'___________________________o T S T c g M O f T *•: V ? V .* * ■.... ' ■ -v Kndolhul Maleic hydratide Growth rolardalion Growtli retardation k Negative oxccpt for slight changes of inanition Negative Aeliva ora q-l'm pyl ¡Homo Pqtcrunyl Imloxidu Van l)ykc 2til All Hlmwed liver nnlargriiifiil of u|>-. All allowed about the Hamo ilegreo of periportal liopalin cell hypertrophy und slight fully chango, and of ruuul tubular pruxImuU-ly tho samo degree pigment of a weur-and-lear typo 10S0 ANTU Wurfurin Zi ne phosphide M ortality; bleached teeth; enlarged Chronic congestion of spleen; atrophy of testis. Teeth not seospleen; atrophic testes tioned M ortality (200 ppm); growth retar* Hyperplasia of spleen, bone marrow and thyroid; thickening of dation; spcctuele eyes; thinning. spongiosa and thinning of cortex of long bones; hyaline cen* trolobular hepatie cells; calcified renal medullary tubular of hair; deformities of legs and casts; terminal focal necrosis of stomach mucosa, thinning of feet adrenal cortex Multiple hemorrhages Multiple hemorrhages; questionable slight inflammatory changes in heart and liver M ortality. Liver damage (discolora­ F atty degeneration and nucrosis of liver tion, nutmeg appearance, rough­ ness of surface) Miscellaneous Arasan M ortality (2500 ppm); disorientation; (unsteady on rear logs) Arumi to Liver tumors and discoloration; growth retardation High m ortality (2500 ppm); growth rolurdution Growth retardation Phygon 5189 3D C Preliminary examination indicates th at histological changes will be slight, except for changes of inanition, in abdominal and thoracic viscera. At 2500 ppm, calcification in brain stem and cerebellum, and dystrophic changes in leg muscles Ilcpntie cell adenomas frequent a t 6000 ppm; hyperplastic changes at 1000 ppm; 200 ppm not yet oxamined Possibly increused incidence of focal nephritis; 2500 ppm uni* mats not examined Unusual. Negativo except for foamy macrophages in lamina propria of villi of small intestine ASSOCIATION O F FOOD AND DRUG OFFICIALS Hoden licldca 6 T 6 T 8 8 M O C J CG0360S 132 ASSOCIATION OF FOOD AND DRUG OFFICIALS L auo, £ . P. and K cnze , F. M .: Effect of carbon tetrachloride on toxicity and storage of methoxychlor in the rat. Federation Proc., 10, No. 1., P art I.t 318,1051. D atidow , B., H aoan , E. C., and R adohsxi, J . L.: A metabolite of chlordane in tissues of animals. Federation Proc., 10, No. 1., P art I., 291,1951. L auq, E. P., K unze, F. M. and P biceett , C. 8.: The occurrence of DDT in Human V fat and milk. Arch. Ind. Hyg. and Occupational Med., 3, 245,1951. ¿J' D atidow, B. and F hawlet, J . P .: Tissue distribution, accumulation and elimination of isomers of benzene hexachloride. Proc. Soc. Exp. Biol. Med., 76,780,1951. 'v^ D atidow, B. and R adomski , R. L .: Metabolite of heptachlor, its analysis, storage and toxicity. Federation Proc., 11, No. 1, P art 1 ,336,1952. F hawley, J . P., H agan , E . C. and F itzhuoh , 0. Q . :A comparative pharmacological and toxicological study of organic phosphates-anticholinesterase compounds. J . Pharmacol. Exptl. Therap., 105,156,1952. Pathology N elson , A. A., et al .: Histopathological changes following Administration of DDT to several species of animals. Public Health Reports, 59, 1009,1944. N elson , A. A. and W oodard, G.: Severe adrenal cortical atrophy (cytotoxic) and liver damage produced in dogs by feeding 2,2-bis-(parachlorophenyl)-l, 1dichloroethane (DDD or TDE). Arch. Path., 48, 387, 1949. by feeding at low levels, 1 to 100 ppm. Federation Proc., 9, No. 1, P art 1 ,339,1950. {j «i ■'* •• GG05G03 5190 Ï6TS Harnessing Chemical Sprays to Serve A the SPORTSMAN through new game management practices Since the beginning o f tim e man has looked at new ways o f doing things w ith suspicion, som etim es justifiably. T h e American sportsman proved no exception w hen he first saw highway departments and power companies doing w holesale spraying of highway and utility right-of-ways to control brush w ith new chemical weedkillers. In many sections of the country, there has been an outburst of vio­ len t criticism of chemical vegetation control practices. T h e spoksemen for the sportsmen have claimed that such spraying destroys gam e, gam e habitat, flowers and natural beauty. W h ile there are certain real hazards connected w ith chem ical weed control, properly used, m ost of the new id killers can successfully be harnessed to serve the ^ jrtsm en through better gam e management practices w ithout any significant evils. It is true that all spraying should be done intelligently and under the supervision of trained, com petent people. T he manufacturers o f weed control chemicals are doing their level best to see that their materials are judiciously applied. Like the sportsman, they condem n spraying which is done promiscuously and indiscrim inately. T he sportsman need have no fear that maintenance men o f highway departments and of America’s power, telephone and telegraph com panies are not alarmed w hen they hear reports of promiscuous spraying, because they are alarmed. T h ey have much at stake. Chemical vegetation control m eans a savings o f m aintenance costs that adds up to m il­ lions of dollars. T hese savings are directly reflected in our utility rates in the case o f power companies. Railroads and highway departments are effecting similar savings. The American farmer was the first to make the use of chemical weedkillers standard field practice. In the grain states he increased phenomenally his output o f small grains and corn by reducing w eed com petition. For the most part, his application of chemicals in crops has not brought objections from the sportsmen. It has always been essential for farmers to cut brush in pastures and in drainage ditches. Like industrial users of spray materials, h e was quick to discover the economies o f chemical controL M ost farmers are anxious to leave cover where it does not interfere w ith sound farming prac­ tice. 3 y selective use o f brush killers woody species that beneneither sportsmen nor farmers are bein g removed from fence rows to perm it desirable species to spread or to be planted if they are not present. In this new field o f habitat control by chemicals the sportsman and farmer have surprisingly parallel interests. The program presents the sportsman w ith a new opportun­ ity to better his public relations program w ith the farmer if he makes him self familiar w ith the farmer's problems and understands their relationship to gam e habitat. T he chemicals m ost widely used for vegetation control are 2,4-D and 2,4,5-T. N either of these chemicals is toxic to man, animals, birds, fish or insects w hen used in accordance w ith recommended practices. Actually, in the case of 2,4-D and 2,4,5-T dosages in ex­ cess o f those com m only employed have been found to have no adverse effect on animal life. Tests have been conducted at several land grant colleges in w hich pastures were sprayed w ith common weedkillers, including 2,4-D and 2,4,5-T w ithout adverse effect on cattle, horses, sheep, pigs, and chickens. In the laboratories of America's chem­ ical industry and in the land grant colleges and in private testing laboratories these materials have been thoroughly studied by toxicologists. 'Wide experience over m illions o f acres has verified these experimental findings. For ex­ ample, 2,4-D has been widely used for water hyacinth con­ trol on the bayous of. Louisiana and in Florida w ithout ad-, verse effects on fish. T he hue and cry against indiscriminate spraying without, any doubt started in instances where spray operators on These men of West Penn Power Company crews are spraying a brush killer which is non toxic to wildlife. The relatively small strip of land they spray will be converted to grass by the second season. Over the years it will save the company's customers millions of dollars. Most conservation people recognize that this spray operation represents conversion of habitat rather thari destruction of habitat. 5192 D o w n to Ea r th , Fall, 1952 y highways attempted chemically to eradicate tail brush badly in need of cutting. It is understandable that the sportsman who saw the un­ pleasant browned appearance that resulted from a few instances of such indiscrim inate and ill advised spraying would w ell b e concerned about the future of w ildlife habitat. Fears commonly expressed in relation to roadside spray­ ing include the danger of erosion and the destruction of habitat. M ost thinking game management m en w ill agree Railroads have a different problem than utility and telegraph companies in that they must remove brush for several yards from the track and all vegetation near the ties. This is a must for safe operation of our nations railroads. Obviously, grazing on railroad right-of-ways is not safe for animals, wild or domestic. that the presence of brush along the roadside for game cover has been overemphasized. Such heavy cover is hazardous to the motorists, to the game, (w hen you consider fast m ov­ ing traffic) and it sets up an ideal opportunity for the poacher w ho shoots from car windows. Actually, there w ill still be adequate roadside cover for sm all game. Grasses grow profusely when brush is removed and more sunlight can reach the soil. Grasses are not damaged by 2,4-D or 2,4,5-T. Thick grass sod is one of the best covers for the prevention of erosion. In Okla­ homa, soil conservationists have found 45% less run-off from a productive pasture previously sprayed to k ill scrub oak than from adjacent m ixed grass-brush land. Dead brush along roadsides after chemical spraying is admittedly unsightly but it is a temporary factor in the process of chemical conversion from brush to grass. Spray crews know now that they must spray brush before it grows too tall or remove it by cutting. Stumps are then sprayed to prevent regrowth. Unless sprayed brush is tall, the in­ creased growth of grass obliterates much of the dead brush. The control of weed vegetation on right-of-ways is an integral part of maintenance. Proper use of selective herb­ icides can serve to convert game habitat rather than to destroy it. W here the outlook is conversion of habitat rather than destruction, game management authorities are beginning to see the value of selective herbicides as new tools. In actual practice, some of the utility companies who are criticized by the sportsman when he first sees discolored brush, may in the long run pay for and provide improved habitat. The game specialists w ith whom the subject has been discussed are generally agreed that the grass land devel­ oped along lines by vegetation control is good for rab­ bits and deer. 2,4-D and 2,4,5-T are ideal for use in game manage­ ment because of their very low toxicity to all animal life and the degree to which accurate application can be controlled. For exam ple, it is possible to k ill selectively susceptible species growing with more resistant species by using marg­ inal concentrations. The Forest Service has used these materials applied w ith hand sprayers to the base o f large hardwood trees to lib ­ erate desirable pine piznrings a t a co st considerably isekrw “that o f hand slashing. These same compounds are used to remove hardwood species from the more resistant Lob­ lolly pines in the southeast : In many areas under today's improved forest fire protec­ tion we have a condition in which w ildlife species h i need of grass and low-growing brush are finding uniform forest stands, such as aspen in the lake states, a very unfavorable environm ent This is a recognized fact and, in areas where there are no desirable trees, game managers already have resorted to the use of the axe, controlled fire and even the bulldozer in an effort to open up the forest crown to perm it growth o f low brush and herbaceous material. W hen w e consider costs o f labor and mechanical bulldozing it is apparent that chemicals can com pete very satisfactorily. Judiciously used, chemical sprays can often do an even better and safer job at this lower cost. Consider the case of controlled burning — for game habitat improvement. Often on poor soils it is several years before any volum e of ground vegetation reappears. In the m eantime w e have an erosion problem and quite possibly we jeopardize our public educational program for forest fire prevention. U sing an aerial application it is possible to kill or injure the overstory of poor trees such as aspen and thus encour­ age the development of low growing shrubs and herbaceous plants. The plausibility of aerial application is proved by die past spraying of 500,000 acres o f m esquite in the south­ west at an average cost of approximately $5.00 per acre to improve carrying capacity of rangeland. W e do not wish to im ply that selective herbicides w ill revolutionize game management practices. The need for interspersion of cover type is w ell recognized for such spe­ cies as sharptail grouse, ruffed grouse, w hitetail deer, cottontail rabbit, and other w ildlife. Chemical sprays are just another tool available to professional game m en in their effort to manipulate environmental factors to speed up production of today’s hard hunted species. Like controlled burning and other established practices, indiscrim inate use of herbicides could prove an enemy of w ildlife. Properly employed, they can be useful tools in the hands of game management people. As new herbicides are developed and carefully tested it w ill becom e increasingly easy and profit­ able to selectively kill worthless plants w hile encouraging the type of cover that may be desired in any given area. Farmers know what chemical maintenance can do to reduce weed population and taxes that are used to support our roads. Every­ where spray crews go there is widespread interest. This picture was taken in Ohio. Such programs can be conducted■without injury to game or adjacent crops. Education and cooperation are the keys to success of such programs. 8 5193 Woody Plant Control Along Roads n Shasta National Forest By A lva G. N e u n s , Training Assistant, California Forest and Range Experiment Station1 A mountain road in the Shasta National Forest mowed in ¡949 and sprayed in 1950. quite another. Manpower shortages make it almost impos­ sible to keep good foremen or spray men for any length of time once they are trained. Consequently, job operations and equipment were simplified as much as practicable. During the 1950 season, then, it was possible to spray 70 miles of previously cut, sprouting roadside brush with a high degree of success and at reasonable cost. The follow­ ing rules for spraying and the slogan, "100% Coverage — 100% Control,” were based on the experience gained that year and are now used as a basis for all crew training.- C o n t r o l l i n g sprouting brush species in the moun­ tains with sprays of 2,4-D and 2,4,5-T depends mostly on how the chemical is applied. This has been demonstrated on the Shasta National Forest in California, where a fight is being waged against roadside brush with the battle cry: 100% C overage — 100% C o n t r o l One hundred and eighty miles of brush were sprayed in 1950 and 1951, and emphasizing the importance of total coverage accomplished three things. First — and most valuable — it gave something to achieve, a definite goal, so that "green’' crews could be trained quickly and easily. Second, it assured complete kill of all sprouting growth above the ground and a maximum of root kill. Third, control was successful and long lasting enough that retreatment the next year was usually unnecessary. The last is important because of the inaccessibility of most forest roads. Shasta National Forest has a network of 1200 miles of forest roads. They wind their way over steep mountain­ Work closeous country from an elevation travel slour of 1000 feet or less to 9000 feet. Some 900 miles of them have brush covered rights-ofway which in the past had to be hand-cut, bulldozed, or mowed with a brush mower every 3 to 5 years to keep the roads from disappearing be­ neath the brush. In a search for less costly brush - control methods, the California Forest and Range Experiment Station tested the selective herbicides — 2,4-D and 2,4,5-T — from 1945 to 1949. Tests were made on plots growing more than 15 troublesome species, including live oaks, deciduous oaks, maples, manzanitas, chinquapin, cherry, snowbrush, and whitethorn. It was found that all of che above ground parts of these species could be completely killed and that the total root kill was high if complete coverage was achieved. Seedlings of all species could be completely killed. H ow to Sp r a y : ( 1 ) Cover all leaf and stem surfaces to ground level with spray— especially the undersides of the leaves. ( 2 ) Travel slowly so that you can see what you are spraying and know when the spray is covering the plant. This usually means walking rather than riding the truck. ( 3 ) Hold the nozzle close to the shrub so that the spraymist will not be blown away or wasted. (4 ) Get down into dense growth using your short boom to pull it apart as you spray. (5 ) Keep pump pressures high enough to ensure spray turbulence and misting. Regrowth like this is ready for spraying. (6 ) Check back after spraying from time to time to find what degree of coverage you are getting. Rule (6) is very important. The spray-crew foreman should check back continuously to make sure all the growth is being treated. W hen diesel oil or an oil and water emul1The California Forest and Range Experiment Station is main­ tained by the Forest Service, U. S. Department of Agriculture, in cooperation with the University of California, at Berkeley, Calif. "The use of 2,4-D and 2,4,5-T for Brush Control on California Roads and Trails. U. S. Dept, of Agriculture. Forest Service, Berkeley, 1950. Chemical spray carefully applied by research workers on plot-sized road shoulders is one thing, but the same formulation in the hands of the average spray man is 14 5195 7 —- sion is used as a carrier for 2,4-D and 2,4,5-T, the spray material can be seen on the leaves and stems of the plants for several hours after the spray is applied. And nothing shows a spray-crew the importance of complete coverage more than to take them back over the job a week or two later when the leaves are drying. The men can see how much is still green when they don’t get coverage — also how dead the plants look when they do. Equipment used on the Shasta National Forest is simple, inexpensive, and readily converted for fire fighting and other purposes. It consists of a stakeside truck geared to travel very slowly and big enough to carry eight, 50-gallon drums—an average day’s supply of spray mixture. A pump used in fire fighting that will develop up to 250 lbs. pressure (100 lbs. is minimum operating pressure) is mounted on a rear corner of the truck bed, 2 high-pressure hoses are attached and fitted with a short boom for average spray­ ing and a longer boom for high banks. Trigger-type shut­ off valves and solid cone nozzles (30° angle spray, 1.1 GPM at 100 psi discharge) on swivel fittings complete the assem­ bly. Pumping is done directly from the 50-gallon drums. A low step is mounted on the back of the truck to carry spray men short distances or while spraying light brush. This equipment is easy to use and requires little main­ tenance; 180 miles were sprayed continuously with no lost time due to equipment failure. Walking speed and trigger control on the nozzles con­ tribute as much to economy of spray as they do to total coverage. Complete control of the spray is a necessity wherever plant cover varies .in density as it does on moun­ tain road shoulders. Dense cover always requires contin­ uous spraying, but medium and light densities do not. In•ermittent spraying — shutting off the nozzle between plants—prevents waste of material on bare ground. Average spray costs in 1950 on 70 miles of road, includ­ ing labor, equipment and materials, were $28.00 per mile. Brush on all road shoulders had been cut and cleared for visibility and reduced fire hazard, with a brush mower or bulldozer, and the sprouting growth was treated with chemical spray. Some mechanical clearing costs as much as $240.00 per mile. Chemical control can eliminate the need for cutting in the future. Re-spraying will be neces­ sary from time to time, but costs should decrease as control ecomes well established. To decrease costs, oil and water emulsions were substi­ tuted for straight diesel oil as a carrier in 1951. Although the average cover was much denser in the area sprayed T h e spray m e n -walk unless there are very high banks to treat. that year, costs on 110 miles of road were $25.00 per mile. As little as 4 gallons of oil per 50 gallons of emulsion were used, but oil was increased as the season advanced to insure better penetration of tougher leaves and stems. During the 1950 field program an attempt was made to vary concentrations of 2,4-D and 2,4,5-T according to whether the predominating woody plant on the site was hard or easy to kill. Since spray material output averaged one 50-gallon drum per mile, it seemed possible to change the formulation in each new barrel to fit the hardest to kill species on the mile ahead. A table was provided from which the foreman could figure the number of quarts of each chemical needed per 50 gallons for a given concen­ tration. Concentrations ranged from 8000 ppm 2,4,5-T and 4000 ppm 2,4-D to 1000 ppm of each for light retreat­ ments. In 1951 to further simplify the job, concentrations were standardized at 4000 ppm 2,4,5-T and 2000 ppm 2,4-D. It was found to be eas­ ier to mix chemical, oil, emul­ sifier, and water at headquar­ ters than depend on mixing and changing concentrations on the job. The slight advantage gained in root kill from higher concentration is offset by the loss of efficiency and labor time involved. Spraying can begin as early as May in the lower alti­ tudes and continue throughout September in the higher altitudes. The season of most rapid plant growth does not occur until late in July in .the high country, so that a long spray season can be planned. Woody plant species, exposure, stage of plant growth, soils, temperatures, moisture and growth rate all change mile by mile on forest roads. These and other factors are thought to influence the root killing power of 2,4-D and 2,4,5-T. But only one influencing factor can be made consistent throughout a mountain road brush-control job. That one factor is coverage, and a well-trained spray-crew and foreman who make total coverage a matter of pride can make the difference between poor control and 100 percent control. V 5196 D own to Earth, Summer, 1952 AH 5197 A+ i Controlling Scrub Aspen W ith BASAL SPRAYS By J o h n L A r e n d , Research Forester Lower Peninsula Forest Research Center, Lake States Forest Experim ent Station, East Lansing, M ichigan in number to satisfactorily restock the stand w ith aspen reproduction. Although the density and vigor o f aspen sprouting can be reduced by summer cutting of the parent tree, the reduction is not sufficient for satisfactory controL B a s a l Sp r a y s T e s t e d : Starting in February 1950, die Lake States Forest Experi­ ment Station initiated a number of chemical herbicide tests in northern lower M ichigan designed to control sev­ eral low value hardwood species, including scrub aspen, oak, and red maple, for pine release purposes. D ifferent methods of chemical control including basal sprays, were applied to these low grade hardwoods in the winter, spring, summer and falL A lso, since aspen sprouting has been found to be somewhat affected by the season o f the year when the parent trees are cut, a special seasonal study was made of scrub aspen treated w ith basal sprays at weekly intervals throughout the entire growing season (M ay through O ctober). B a s a l SPRAYS, also called "bark and dormant sprays,” for controlling undesirable woody plants have been under study by various investigators in M ichigan since 1946*. Ib is paper describes the results of basal spray tests made three years ago which appear to offer a satisfactory method for controlling scrub aspen in northern lower M ichigan. The basal sprays tested included esters of 2,4-D (Esteron 44 and Esteron T en-T en), 2,4,5-T (Esteron 2 4 5 ), and 5 0 /5 0 mixtures o f each (Esteron Brush K iller) m ixed w ith Grade 3 diesel oiL Concentrations of 2 and 4 percent by volum e (8 to 16 lbs. o f acid per 100 gals.) were applied to T h b C o n t r o l Pr o b l e m : Aspen is a valuable tree species for wood products on good forest sites in the Lake States region. However, on poor sites aspen is usually an inferior grade tree. As a result, forest and game managers frequently need to control scrub aspen to improve w ildlife habitat or establish tree species m ore adaptable to the forest site. W hen aspen trees are cut or girdled mechanically, numerous sprouts and suckers develop from che stumps and roots for distances as far as 100 feet from the parent tree. Consequently, aspen reproduces itself very w ell after cutting. T o date no satisfactory method for reducing the sprouting o f scrub aspen has been reported. Several investigators ** have reported that the season of the year when aspen trees are cut has som e effect on the vigor and number of sprouts and suckers which develop. Sprouts developing from aspen trees cut during the summer months have been found to be less numerous and less vigorous than those developing from trees cut at other seasons o f the year. Aspen sprouts developing from trees cut in the summer, however, have generally been sufficient 1At Dow Chemical Company, Midland, Michigan, and Michigan State College, East Lansing, Michigan. ' Zehngraff, P. J. 1946. Season cutting affects aspen sprouting. Lake States Forest Experiment Station Technical Note 230. *Stoeckeler, J. H. 1948. The growth of quaking aspen as affected by soil properties and fire. Jour, of For. 46(10) ¡727-737. Aspen sprouts and suckers which developed from aspen trees treated with -basal sprays during the dormant and earh growing season. 10 at various seasons: ( 1 ) dormant (February); (2 ) early growing season (M ay); (3 ) summer (Ju ly); and (4 ) fall (Septem ber). The sprouting results on the other test areas were the same, L e., no sprouts or suckers were found among the scrub Aspen rrearea in July and only a few among the aspen trees treated in September. However, sprouts were both numerous and vigorous among the trees treated in February and in May. the basal portion of individual trees to heights of 2 and 4 feet. In all cases the basal portion treated was carefully w etted to produce-runoff com pletely around the tree. r E c r s o f B a s a l Sf r a y s o n T o p s o f Sc r u b .A s p e n : A ll of these ester formulations o f 2,4-D and 2,4,5-T when m ixed w ith diesel o il at concentrations o f 2 to 4 percent by volum e and applied to die basal portion of aspen in su ffic ie n t q u a n tity are capable of k illin g the top of the tree. T he volum e o f herbicide applied in basal spray work is very important, especially for the larger trees. For example, only slight differences were observed in the rate of top k ill and subsequent sprouting o f aspen treated w ith 2 and 4 percent mixtures of esters of 2,4-D and 2,4,5-T in diesel o il when the basal portion of the trees were sprayed to a height of 4 fe e t Moreover, a 2 percent mixture applied to the basal portion of the trees to a height o f 4 feet is consistendy more effective than a 4 percent m ixture applied to a height o f 2 fe e t H owever, spraying the base o f the tree to a height of 4 feet requires a considerable volume of herbicide compared to a 2 root h eig h t S u m m a r y a n d C o n c l u s io n s : Scrub aspen trees have been killed without subsequent sprouting and suckering for three years in northern lower M ichigan by applying basal sprays during the summer months of late June, July, and August follow ing full leaf developm ent. Aspen similarly treated during the dormant and early growing seasons suckered prolifically beginning the secon d year after the basal spray treatment. Season of the year during which the scrub aspen is treated with basal spray has considerable effect on the concentra­ tion and volume of spray applied for an effective top k ill as w ell as subsequent sprouting of the treated trees. A 2 percent mixture (by volum e) of 2,4-D and 2,4,5-T (Esteron Brush K iller) in diesel o il sprayed on the basal W hen scrub aspen is treated w ith basal sprays (2 to 4 percent mixtures by volum e of 2,4-D and 2,4,5-T esters in diesel o il) during the dormant season, the tops are gen­ erally dead by the end of the first growing season follow ing the treatm ent The treated trees usually leaf out in the spring, but the foliage starts turning yellow shordy after full leaf developm ent Scrub aspen treated w ith basal sprays during the early grow ing season starts showing the effects o f treatment in about 10 days. However, trees treated in late July and August probably w ill not show the effects o f the basal spray treatment until the follow ing growing season. In other words, during the early grow­ ing season the tops of aspen trees are more easily killed by basal spray treatment than they are at any other season of the year. D uring the late growing season, and in the fall and w inter months, aspen trees are generally more resis­ tant to basal sprays. It is during these seasons that 2,4,5-T esters are more effective than 2,4-D esters. Also, a larger volume o f herbicide needs to be applied to the basal portion of the tree during the fall and-winter months than during the early grow ing season. Consequendy, a 3 percent m ix­ ture by volum e o f Esteron 245 (2,4:5-T in diesel oil applied from d ie ground level to a height o f 2 feet in soaking quan­ tities is an effective year-around basal spray treatment for woody plant control. E f f e c t s o f B a s a l S p r a y s o n S p r o u t in g o f A s p e n : N o sprouts or suckers were found at the end of the first g ro w in g season from the aspen trees treated w ith basal sprays at different seasons of the year. However, those cut and girdled sprouted prolifically the first year— the sprouts averaging about 3 feet in height. A t the end of the second growing season suckers were found w ithin 2 to 5 feet of the basal-sprayed aspen, e xce p t fo r th ose treated a fte r fu ll lea f d evelo p m en t. T h re e g ro w in g seasons a fte r the basal spray treatments the same sprouting condition prevailed — no suckers or sprouts were found from aspen treated after full leaf developm ent through September, whereas suckers developed from the aspen sim ilarly treated during the dor­ mant period and during the early grow ing season before full leaf development. Aspen trees killed by basal sprays applied during late June, July and August after full leaf development. No sprouts or suckers have developed for three growing seasons after the trees were killed with basal sprays. (Trees sprayed summer, 1950. Photo taken in September, 1952). portion of the tree to a height of 2 to 4 feet is effective in killing the top of aspen during die growing season. How­ ever for year-around basal spray treatment, a 3 percent m ix­ ture of 2,4,5-T (Esteron 245) or 12 lbs. of acid equivalent per 100 gallons; o f Grade 3 diesel oil, applied to die basal portion of the tree to a height of 2 feet is recommended. Basal spraying aspen w ith either o f these herbicide m ix­ tures during late June, July and August follow ing full leaf developm ent offers a promising method of controlling scrub aspen w ithout subsequent sprouting for at least three years. H ie weekly basal spray tests were conducted only on one area of scrub aspen. However, sim ilar basal spray treat­ ments were also applied to scrub aspen at tw o other loca­ tions in northern lower M ichigan during the same year 11 _ 51 D ow n to TH, Summer, 195 008S .DOWN to EARTH 1 T ^ ^ C o p y r ig h t I Q 5 3 by The D o w Chemical Company J-IW A review of agricultural chemical progress volume 9 no 3 winter 1^53 Chemical Control of Woody Plants in C A LIF O R N IA By O liver A Leonard , Associate Botanist, University of California, Davis, California Ribes sp. must be controlled in the sugar pine (P irns lambertiana) sites, since many members of this genus are alter­ nate hosts for the white pine blister rust. Willows along stream banks hinder streambed maintenance and thus make flood control more difficult and complicate the mosquito control problem. Chemical brush control along rights-ofway is becoming more common. Poison oak (Rhus diversiloba) is of personal concern to more people in California than any other woody plant pest. It is controllable with either 2,4-D or 2,4,5-T, although 2,4,5-T is slightly super­ ior. From one to three applications, spaced at yearly inter­ vals, have been necessary for a complete controL There is some evidence, too, that acid or amine formulations, plus 1 % low toxic spray oil, may result in better control than is obtained with some esters. The Botany Department, University of California, Davis, California is conducting basic research on the chemical con­ trol of woody plants, including ( 1 ) cuticle penetration of herbicides and ( 2 ) radio-isotope studies. Working with live oak (Quercus wisUzenii) and toyon (Pbotinia arbutifolia), for example, it was determined that 2,4-D moved rapidly out of treated leaves and thence downward in the bark in February when the shoots were still "dormant.” Growth of roots during this period related both to the downward flow of materials from the leaves and to the fact that these woody plants are sensitive to hormone sprays at this time of the year. Some general results of experimentation in chemical weed control in various types of woody vegetation are presented. B otanist O liver A . Leonard on the D avis Campus o f th e U niver­ sity o f C alifornia p oin ts out 2,4-D filled cuts around th e base o f a large live oak tree. T h e etas go an inch or tw o in to th e wood. W oody plants constitute problems on some of the follow­ ing areas in California: ( 1 ) 2,500,000 acres of commercial timberland that is little more than brush; ( 2 ) 7,500,000 acres of woodland-grass; ( 3 ) 7,300,000 acres of chamise; ( 4 ) 2,400,000 acres of chaparral other than chamise; ( 5 ) 5,000,000 acres o f big sagebrush. "Woody plants, also, are problems on some other areas. A map is presented showing the distribution of some of the. more important areas where brush is a problem in California. There are many groups of people whose welfare is involved in controlling, woody plants. Ranchers are interested in brush control in order both to reduce brush encroachment on cleared rangeland and to reclaim valueless brushland by transforming it into good pasture. They are interested in controlling brush and trees around springs and along water courses in order to have more water available for livestock and domestic purposes. In some areas, there is interest in controlling "woody plants to make more water available for irrigation. Foresters are interested in woody plant control to aid in reforestation. Scotch broom ( Cytisui scoparius) is a serious pest in the forested areas of several Sierra Nevada counties and prevents natural pine reforestation. It is readily killed by one application of 2,4-D (low-volatile ester) applied as a foliage spray in June or July, but seedling problems have not been solved. Coastal Sagebrush a n d Coyote Brush A reas : Coastal. sagebrush (Artem isia califomica) and coyote brush (Baccharis pilularis) are readily killed with 2,4-D at rates of 2 to 3 pounds per acre, applied by aircraft. Accord­ ing to available evidence, results should be more uniformly satisfactory when amine 2,4-D is used. The recommenda­ tion is 3 pounds acid equivalent per acre in about 8 gallons of water and one gallon of light-medium summer spray o il The favored period for spraying is during April, May and June, but good results have been secured on coyote DOWN TO EARTH * A R eview o f A gricultural Chemical Progress Published by T h e D ow C hem ical Company, • ON THE COVER • M idland, M ichigan, m anufacturers o f agricultural chemicals. Tw enty-one thousand copies distributed to those engaged in agricultural research and instruction in Am erica and abroad • Eugene Perrin, Editor. IN THIS ISSUE • C hem ical control o f woody plants in California • A sim ple device fo r applying E steron. 245 * E thylene dibrom ide fo r th e control o f nem atodes in th e organic soils o f M ichigan • D alapon, a n e w . system ic grass killer introduced fo r control o f industrial vegetation • T im ber stand im provem ent on the W . G. Jones State Forest, Conroe, Texas • R eport on Venezuelan experim ents on grass control in rice w ith Premerge. See picture legend on P*g* 3 2 5202 brush up to September 1. Two pounds o f 2,4-D ester (low -volatile), plus 1 gallon of Diesel oil and.8 VS gallons o f water per acre have resulted in kills only slightly in­ ferior to chose obtained with the amine treatment. Control of mixed coastal brush with one application by aircrafc is not adequate and must be followed by some addi­ tional spraying, probably with a ground rig. A t present, the iniual spray is with a brush killer using 2 pounds of acid equivalent per acre, plus 1 gallon of Diesel oil and 8 Vi gallons of water, applied by helicopter. The advantages of spraying over other methods of brush control have been outstanding, especially in terms of in­ creased grass production. C h a pa r r a l a n d C ha m ise A reas : Because chaparral and chamise occupy so much land in California, considerable study has M en undertaken to determine feasibility of control methods, the pattern of recovery from various treatments and the performance of desirable associated vegetation and subsequent seeding. Chamise and chaparral have frequently been burned in the past, but these areas usually revert rapidly to the origi­ nal condition because of crown sprouts and seedlings. The sites are frequently almost devoid of grass before being burned and killing the brush is of little value. Grass can be established most readily by burning the brush in the fall and seeding the desired grasses and legumes immediately. Grass competes with the brush sprouts and seedlings and results in the death of many of the latter. A broadcast spray should not be applied until the second spring after the burning and seeding so that ( 1 ) the legumes will have gone to seed once, building up a reserve of hard seed in the C hem ise urea seeded to perennial grasses. T h e author is standing on th e boundary betw een a sprayed ( background ) and unsprayed ( foreground) area. T h is is the same general area fro m w hich th e data presented in th e table w ere obtained. D r. Leonard is pointing to the location w here 2,4-D am ine bat been placed in a cut in this interior liv e oak 33 m onths pre­ viously. T he wood is rotting and stem s have broken over. E ight separate cuts and 16 m l. o f undiluted 2,4-D am ine were used. soil and ( 2 ) sprouts will have developed and most of the brush seeds will have germinated. Good success has been achieved in killing brush sprouts and seedlings developing after a fire on chaparral ana cha­ mise areas by applying a broadcast spray, consisting of 2 pounds of low volatile 2,4-D per acre in 2 gallons of Diesel oil and 31 Yi gallons of water. March and April appear to be the optimum time for spraying. Such a spray will kill most of the chamise ( Adenostoma fasciculatum), yerba santa ( Eriodictyon californicum) , golden'fleece ( Haplopappus arborescent) , deer-weed (Lotus scoparius), rush rose ( Heliantbemum scoparium), creeping sage (Salvia sonom ensis), and brush seedlings, including chamise, Ceanotbus sp., and Arctostapbylos sp. Seventy percent of the toyon (Pbotim a arbutifolia) may be killed. A partial top kill of interior live oak (Quercus w islizem t), coffeeberry (Rbam nus californica), and redbud (Cercis occidentalis) is se­ cured. The latter species can be killed by from one to chree applications of a spray containing 4 pounds acid equivalent of a brush killer in one gallon of Diesel oil and 98 gallons of water. Where C ea n o tb u s sp . are abundant, there is some advan­ tage in using a spray that contains some 2,4,5-T, especially on the larger plants. Aircraft spraying has not been sufficiently successful thus far to be used commercially. Early in these studies, the isopropyl esters were com­ pared with the mixed propylene glycol butyl ether esters of 2,4-D and 2,4,5-T, when formulated the same. One such test conducted in March 1951 resulted in a 22% and 20% kill of chamise sprouts with 2 pounds of the isopropyl esters of 2,4-D and 2,4,5-T, respectively, and a 78% and 54% kill with the propylene glycol butyl ether esters. The advantages in using some oil in the spray mixture have been demonstrated. There is some evidence to indicate that an oil of low toxicity gives a superior kill to that obtained with Diesel oil on chamise (high volume spraying). There has been an increase in die production of grass on areas that have been sprayed, over that on unsprayed areas. One example of this was on a chamise site. The brush was broken down with a bulldozer in the summer of 1950 and the area burned in October of the same year. Immediately following the burning, the area was seeded by airplane to harding grass (P h a la ris tu b e r o sa ), smilograss (O ry zo p sis m ilia c e a ), tall fescue (F estu ca a ru n d in a c e a ) , and rose 5203 D o w n to Ea rth , W inter, 1953 the sap-wood Trees are sensitive at all times of the year, but less chemical is required to kill trees in the winter and spring than in the summer and falL On Digger pine ( Pinsts sabiniana),. ihe cuts can be 8 inches (center to center) and good results obtained using 2 mL of 2,4-D amine ( 4 lb. acid equivalent) per cut (undiluted). On Blue oak ( Quercus douglasii), the cuts can be 6 inches center to center and again 2 mL per cut should be used Live oak is a vigorous sprouter ana the cuts should be con­ tinuous for best results. Stumps can be treated the same as the standing trees, although some retreatment will be neces­ sary; for example, from one to 6 sprouts had appeared on 4 out of 18 live oak stumps that had been treated 2 years previously with the amine of 2,4-D. 2,4,5-T amine ap­ peared to be rather ineffective, since most of the stumps developed sprouts. Big Sag ebru sh : Sagebrush ( Artem isia tridentata) is being controlled ef­ fectively with a mixture containing 2 lbs. of 2,4-D ester, plus V i gallon o f Diesel oil and 9 gallons of water, applied in June. T his helicopter (.coming in fo r a landing ) -was used to spray coyote brush.' . > clover ( T rifolium b irtu m ). Broadcast. spray treatments were applied in 1932, (note table for one example). The quick returns as a result of spraying are indicated. It should be cautioned that the results obtained in seeding this peren­ nial grass mixture were unusually good. The effect of 2,4-D on chamise sprout kill and on grass production one year after spraying is shown in the table. A broadcast spray was applied on April 17, 1932, using the propylene glycol butyl ether .ester of 2,4-D in 2 gallons òf Diesel oil and 37VS gallons of water per acre. Grass production was determined in July 1953. (Coop, study with Charles Carlson, Calif. State Division of Forestry). : 2,4-D per acre K ill o f Cbamise sprouts D ry w eight o f grass per acre lbs. % lbs. 0 1 2 0 50 87 639 1378 1771 Co n c l u s io n s : Many different problems exist on the millions of acres of brush covered land in California. Ch<»miraU are being used in many situations and their use will increase since certain jobs can be done better. However, on much of the low-value rangeland, it will be necessary to do a good job more economically than is now possible. This is a challenge to industry as well as to federal and state research and extension agencies. & J: W oodland -G rass A rea s : There is considerable interest in clearing some of the woodland-grass areas. These areas are often burned, but a top kill is usually not obtained, except in live oak thickets or where brush has invaded. 1.Bulldozing is commonly employed, especially where the soil and topography are suitable. After bulldozing and after fire, live oak sprouts develop from underground parts and grow rapidly, so that within a few years die area reverts again to the original state. The present recommendation is to spray the sprouts with a mixture containing 4 lbs. of 2,4-D plus 2,4,5-T (Brush Killer), plus one gallon of Diesel oil plus 98 gallons of water. The spraying should be in the spring and early summer months and repeated until the sprouts, are all killed. Very few sprouts will be alive after the third year. The best kill of live oak sprouts thus far achieved by one application was with a spray containing 2 lbs. of 2,4-D (amine form) in 100 gallons of water plus .1 pint of a sticker spreader. The spray was applied in October of 1950 and by July of 1953, 56% of the stumps had no live sprouts left. The cut-surface method using the amine of 2,4-D is ap­ plicable to the oak woodland areas. Cuts made with an axe or hatchet should be deep enough to extend well into V:? ir?-'• < _ v-y. !■ ■ 4 5204 w A Simple Device for Applying Esteron 245 By DAVID B. COOK Cooxrox Forest, Stepbentown Center, New York S'*«"" — w-»-n->JTw.tact ■ r EDITOR’S NOTE: Mr. Cook reports on a home-made variation of basal bark application equipment. Two years’ use has convinced him of its practical value. W e are glad to publish his ideas which may prove helpful to others. ESTERON 245 is rapidly making a name for itself as an effective silvicultural tooL Mixed with water, it can be used advantageously as a foliage spray on woody plants up to eight feet talL Trees larger than that, and up to 6 inches in diameter at the ground, can best be treated with a basal spray of ESTERON 245 in oiL This latter technic is particularly useful in the selective deadening of unwanted, unmerchantable stems where a residual stand is to be re­ tained. It is also a means of treating cut stumps to prevent sprouting. The trade literature suggests that basal sprays be applied with a knapsack or small tank sprayer equipped with a wand bearing a nozzle set to deliver a narrow, solid cone of spray at low pressure. Such equipment is quite expensive. Moreover, it is impractical to use it for anything but "hor­ mone” sprays because of the difficulty of completely clean­ ing it, and because "hormones” are effective against certain sensitive plants even in very small amounts. And, in prac­ tical operation, it is difficult to maintain an even, low pressure and to keep most of the spray on the target. My early experience convinced me that such rigs were too complicated at least for my use; that pressure was un­ necessary; and that simple gravity flow would be sufficient. It was apparent that a free flow of mixture would give a more thorough soaking effect and would run down onto the thin bark a t the root crown better than would even the coarsest pressure spray. A simple and effective basal sprayer was assembled from a two-gallon lubricating oil can with an air vent punched in the cap and a short pipe tapped into the bottom. For convenience in carry­ in g , t h e can w as mounted on a ply­ wood packboard with rope shoulder straps. T o the can was at­ tached three feet of Y&" "pure latex tub­ ing”, such as is used for handling blood. (I learned the hard way that ordinary red rubber tubing disin­ tegrates from contact with kerosene). On th is w as threaded a clip-type tubing damp and the end slipped onto a 24" length o f Va " alumi­ num pipe. Valve and aluminum pipe are Basal spray applicator made w ith mounted in a wooden 2-gallon o il can. T h e applicator in use. T h e drenching stream gives the stem a good soaking. handle to give a better hand hold. The whole rig weighs 3 V* pounds and can be put together for a little labor and about $1 in cash,- the latex tubing being the most expen­ sive item. In operation, die tip of the wand is placed against the desired spot on the lower stem, the valve undipped and die thumb pressure released enough to give the desired flow. The bore of the tube is Y&", so the liquid flows out in a solid stream about the size of a match-stick, at no great ressure and with almost no wasteful spatter. Rate of ow can be nicely adjusted by thumb pressure. On trees up to 2" in diameter, the kerosene will spread readily all the way around the stem from a single point of application. Somewhat bigger stems will require some lateral movement of the wand, while those over 4" must be spotted from opposite sides. In moving from tree to tree, the valve can be clipped shut; die small bore of the tube will keep the liquid from running out. W hen not in use, the wana is carried in the dothespin clip on the board, ’ leaving the worker with both hands free. This simple rig has several advantages. There being no pressure to maintain, the operator has one hand free and nothing to think about except aim ing the wand. H e has but to shrug his shoulders to find out from the sloshing how much liquid the can still contains. Filling is simply and quickly done. In two seasons of work, it has proved it­ self a good tool. The formulauon used has been a 4% solution of ESTE­ RON 245 in kerosene, plus Vi teaspoon of R ed-0 oil dye; when freshly applied, this bright red is conspicuous, espedally on grey birch. W ith this device, I have been able to accurately apply 2 gallons per hour, that being enough to treat about two hundred 3-4" trees. It is not so effident on stems less than Vi" in diameter because of the difficulty of holding the wand tip on such a small target, and letting out so small a quantity of liquid. The drenching stream gives the stem a thorough soaking, which may be one of the factors contributing to my uniformly good success in getting complete kill on a wide variety of northern hard­ wood species. 5 5205 D o w n t o Ea r th , W inter, 19- TIM BER STA N D IM PROVEM ENT O n the W. G . Jones State Forest, Conroe, Texas B y C harles T. Stbalby , J r.* inches. The sizes of the stems sprayed ranged from less than one-half inch to eight inches, with the average about four inches. One gallon of the spray solution treated approximately seventy-five stems. The number of stems treated per acre by girdling ranged from ten to thirty, with the average of about fifteen. The number of stems treated by spraying ranged from seventyfive to one hundred fifty, with the average of about one hundred ten per acre. To check die results of the treatments, a cruise was made of - the compartment in November 1952, dividing the treated stems into four degrees of results: dead, dying, ab­ normal and not affected. Table 2 is a summary of the results of the treatments, showing the number and percentage of stems treated, the total number of stems and the cost per stem treated. Tables 3 and 4 show the results of the individual girdling and spraying treatments. The project from the standpoint of releasing established pines was very beneficial A total of ten thousand four In many .areas of the south the once pure stands of south­ ern pines have undergone changes, so that today the under­ story and in many cases the overstory is a mixture of south­ ern pines and southern hardwoods. T o help insure and in­ crease survival, to enable more rapid growth and better development, to increase the maximum financial yields and to remove low valued hardwood competition from the higher valued intolerant pines, various individual treat­ ments and combinations of treatments have been used in timber stand improvement programs. Several examples of individual treatments are girdling, frilling and felling. A combination treatment could be any of these individual treatments and the addition of a chemical herbicide. . The W . G. Jones State Forest, under the administration of the Texas Forest Service, is dedicated to demonstrational and research use. In an effort to obtain cost records and to set up a project as a demonstration of timber stand improvement and research with 2,4,5-T, a combination treatment of girdling and basal spraying with 2,4,5-T was used on a ninety-nine acre compartment of the forest. The compartment treated was typical of many East Texas forest stands. The pine-hardwood overstory was composed of loblolly and shortleaf pine, several species of red and white oaks, sweet and black gum, mockernut hickory, American elm and dogwood, with pine making up sixtyfive percent and hardwood thirty-five percent of the stand. H ie understory was composed o f seedlings and saplings of hawthorns, bays and yaupon in a seventy percent hard­ wood— thirty percent pine—proportion. The initial treatment consisted of girdling all hardwoods, eight inches d. b. h. and larger, which overtopped and/or interfered with established pine trees or reproduction, using the two hack girdling method. After the girdling was com­ pleted, the compartment was treated using the basal spray method. A ll hardwoods less than eight inches d. b. h. that overtopped and/or interfered with the pine were treated. A two and eight-tenths percent concentration, by volume, was used for the basal spray, by mixing one and one-half gallons of Esteron 245 with fifty-three and one-half gallons of diesel oiL This concentration was selected on the basis of research work conducted by Ray E. Goddard, Assistant Silviculturist of the Texas Forest Service, who found satis­ fying killing results were obtainable with it, on the basis of preliminary work, and that it was more economical than the higher concentrations. Spraying began in late March, 1952, and was completed in the latter part of April, using local unskilled labor. An inspection or the results of the treatment was made in November, 1952. Table 1 is a summary of the cost study of the individual treatments and of the entire project. The sizes of the trees girdled ranged from eight to eigh­ teen inches d. b. h., with the average tree about twelve 12 5206 L ooking up a t p in e saplings and dead branches o f overtopping hardwoods. hundred ninety-four trees were released; seven thousand one hundred twenty-eight by spraying and. three thousand three hundred sixty-six by girdling. From the initial observations o f this project it appears that the com bination treatment gave more complete results in timber stand improvement than would either method by itself. Although the costs per acre for the individual treatments were lower than that o f the combination, it only cost an additional one dollar and eighty-nine cents to re­ lease an additional seven thousand one hundred twentyeight trees by spraying, after the compartment had been girdled. T o girdle or lop this smaller material would have m artially increased the girdling cost, not to- mention the probability o f sprouting, if no chemical was used. To spray the larger trees instead of girdling would increase the TABLE I Summary o f C o st Study T im ber Stand I m pr o v e m e n t P r o je c t Cost Ite m '— H ours . C ost/A cre 93.0 $ 69.75 Girdling la b o r ------------27.90 Girdling supervision____ 42.5 Total girdling cost---------$ 97.65 $ 0.99 79.0 Spraying lab o r----------- — $ 59-25 21.76 32.0 Spraying supervision-------28.16 Diesel oil (220 g a L )----78.60 2,4,5-T (6 g a L ) ----------Total spraying cost_____ - $187.77 $ 1.89 Total girdling c o st_____ $ 97.65 187.77 Total spraying c o s t____ $285.42 ' $ 2.88 Total cost ___________ spraying cost materially above the cent and one-half per stem, hence higher costs per acre. The results of this project show that over ten thousand pine trees were released at the nominal cost of two dollars eighty-eight cents per acre by using a combination treat­ ment of girdling all hardwoods over eight inches d. b. h. and spraying all hardwoods under eight inches d. b. h. that were over-topping and/or interfering with established pine reproduction. TABLE II Su m m ary o f R esu lts o f T reatm ents 1 A cre Item 99 A cres Total hardwood stems - _____________ 868 - 85.932 Total hardwood stems treated__________ 136 15,444 ' Percent of hardwood stems treated_:____ 15.6 15.6 Total hardwood stems sprayed__________ " 121 11,979 ; Percent hardwood stems sprayed_______ _ 13.9 13.9Cost per stem sprayed__________ _____ : $.015 : $.015 Total hardwood stems girdled______ - ___ - 15 ‘ 1.485 Percent hardwood stems girdled______ 1.7 1.7 Cost per stem girdled_________________ $.065 $.065 . ' TABLE HI Sum m ary o f Spra y in g T r ea tm en t T im ber Stand I m pr o v em en t P rotect Item Total hardwood stems sprayed__________ Total dead _________________________ Percent d e a d _____ ______ ___________ Total dying _______________ ________ Percent d y in g _______________________ Total abnormal______________________ Percent abnormal ______ ’________ ____ Total no effect______________________ Percent no effect_____________________ 1 Acre 99 Acres 121 101 83.5 5 4.1 15 12.40 0 11,979 . 9,999 83.5 495 ‘ 4.1 1,485 12.4 0 0 1953 O bservations o f R esu l t s : In the spring of 1953, several weeks after the vegetation ■» had leafed out, an inspection was made of the results, Many o f the treated stems which had been reported as 5207 D o w n t o E arth , W in te r, TABLE IV a low percentage concentration, by volume, of 2,4,5-T is sufficient to kill or seriously affect the overtopping smaller hardwoods in timber stand improvement work in-.south­ ern pine-hardwood stands. ' -. Su m m a r y o f G irdling T r e a t m e n t T im ber S ta n d I m pr o v e m e n t P r o je c t Item ________________________ 1 A ere 99 A c rti Total hardwood steins girdled___________ 15 • 1,485 Total d e a d ---------------- 1--------------------14 1,386 Percent d e a d -------------------------------------- 93.3 93-3 Total dying__________________________ 0 0 Percent d y in g -----------------------------------0 0 Total abnormal_______________________ 1 99 Percent abnorm al_____________________ 6.7 6.7 Total no effect ______________ !________ 0 0 Percent no effect______________________ 0 0 Su m m a ry : The results show that after eighteen months, over fifty dead the previous fall had leafed out over one-half of the crown surface. A much smaller number had stump sprouted; had partially leafed out in the crown; or had leafed out from the stem. Over fifty percent of die stems treated showed no visible effects of die spraying what­ soever, and had seemingly recovered. A late summer inspection of the compartment showed chat many of the trees which had seemingly recovered at Pine sapling (foreground ), dead sw eet gum ( im m ediately behind p in e ), and dead oak and sweet gum at le ft. Leaves are dead on tree at le ft. TABLE V C om pa riso n o f Spraying R esu lts T im ber Sta n d I m pr o v em en t P r o tect ' PERCENTAGES _________ Percentage dead __________ Percentage dying _________ Percentage abnormal____ __ Percentage no e ffe c t _______ ____ -Ite m Pall 1952 Spring 1953' Sum m er 1953 4.1 12.4 0 31.4 1.6 14.0 53-0 52.2 38.2 4.0 5.6 percent of die treated stems are dead, in many cases already fallen. Another thirty-eight to forty percent should be dead within a year. A portion of these stems may leaf out again next spring, bur this summer’s results show that only a relatively few will be able to survive and. re: cover completely. In the final analysis, if the results follow present indica­ tions, over ninety percent of the overtopping smaH hard­ woods in southern pine-hardwood stands can be adequately controlled in a timber stand improvement program which is economical and easily accomplished and which produces the desired results. T reating girdled hardwood w ith botai spray. the time of the spring inspection, had died or were dying. Some few had recovered and showed litde visible evidence of any spraying effects. Sweet gum made up over ninety-five percent of the stems which had leafed out and had seemingly recovered in the spring and late summer inspections, and one hundred per­ cent of the stems which showed no spraying effects in the late summer inspection. The red oaks made up the greatest proportion of the remaining five -percent, with only minor percentages for all other species. The following table shows a comparison of the results of the spraying treatment at the various inspection periods. From the three observations of the results it appears that • Formerly District Forester, District 6, Texas Forest Service, Ginroe, Texas. At present General Manager and Forester, Indepen­ dent Pulpwood Producers, Inc., Geveland, Texas. 14 \5208 - * \ > ued. On to r of 13 day3 d those died he« e litter ü dustrv as a fodder supplement for beef cattle, thereby improving’ meat quality and increasing quantity in less time with less food. R cferrn c ei 1. S m a ll , J . IC. T h e n o r m a l o f S o u th e a s te r n F lo ra . Now Y o rk : T u b l. by a u th o r (1 0 3 3 ). li. F e e r t , S. D., a n d F o x , L . E . J . A m . P h a r m . A s s o c .. 41, 433 (1 0 5 2 ). 3. P i n c e s . G „ a n d T h im a n n . K . V. T h e H o rm o n e s , V ol. 2. N ew Y ork : A c ad e m ic P re s s (1 0 3 0 ). 4. W’E lciiE nT , C. K .r a n d K o rb ig a n , S. J . E n d o c n n o Jo i/i/, 5. 741 (1 0 4 2 ). 3. Z o n d e k , B r L a n c e t, lO , 10 (103G ). *». S l e e t i i , R.\ B. A g ric u ltu r a l D e p t., U n iv e rs ity o f F lo rid a , p e rs o n a l c o m m u n ic a tio n . 7. A n d r e w s , F . N\, B e e s o n , W . M., a n d J o h n s o n , F. P . J . A n im a l S c i.r 9 , G77 (1 0 3 0 ). 8. IY eodeb , M . G.. L a c t e h . U \ M., a n d F o o t e , P . A . J . A m . P h a r m . A s s o c ., 41, 2 3 0 (1 0 5 2 ). 9. I I a l l io a x , J . E . J . In d . E n g . C h em ., 1. 2 06 (1 9 0 0 ). 10. P i c k e t t , J. .\L F lo r id a E x p t. S ta . B u ll. I T (1 8 0 0 ). 11. T r e n c h , R . B., a n d A bb o tt , O . D . F lo rid a A g r . E x p t. S ta . T e c h . B u ll. N o . 4J4 (1 0 4 8 ). determination as described by Triiog and Meyer (1). Four hours after trratment the sprayed plants showed epinasty and otlier 2.4-D elTeets. Ten hours after treatment many of the leaves had curled, and the plants were becoming recumbent. The next morn­ ing the plants were somewhat chlorotic and the dis­ tortion had increased. The following day many of the leaves had developed necrotic spots. One week after treatment the leaves on most sprayed plants had withered and those that adhered were very chlorotic and sickly in appearance. The uprooted plants remained fresh for the first day but after that they deteriorated so rapidly that by the end of the week it was not possible to obtain leaf samples. ■ ; Figure 1 shows the fluctuations of inorganic phos L S 0-5V 3O 9 2.4- D Affects Phosphorus M etabolism A. J. Loustalot, M. P. Morris,. LS0-5S.'ia6 I V 224 J. Garcia^ and. C. Pagin. F ederal E x p e rim en t S tation- in P u e rto R ico ,1 td a y a g u e z . A preliminary experiment in which Commelina sp. and Xanthosoma sp. were analyzed 24 hr, and 1 wk after being treated with.2,4-D (2,4-dichlorophenoxyacetic acid) showed that the percentage of water-solu­ LSD-5T.-274 CV 329 ble phosphorus in treated plants was consistently higher than in untreated plants. The following ex­ periment was carried out to obtain additional infor­ mation on the effect of 2,4-D on phosphorus metabo­ lism. H arm t time offer freofmtrtf (Horn) A prepared field, was divided into 12 plots each P ic. I . Levels of in o rp m le phosphorus (ppm o f d ry m at 5 2 x 2 4 f t and planted to a variety of white beans, te r) In bean p lan ts analyzed a t various in te rv a ls s f t e r tre a t Blanca Bonita (P.R. 1632).. The experimental design m e a t w ith 0.1% solution of sodium 2.4-D. consisted of 3 treatments each replicated 4 times in phorns in the leaves, stems, and roots, respectively, of . randomized'blocks. When the plants were about 15 in. high and had treated, check, and uprooted plants. The data obtained at each sampling period, ex started to set fru it 1 plot in each of the 4 replications was sprayed with OJ.% aqueous solution of sodium pressed as parts of inorganic phosphorus per million 2.4- D. The plants in another plot of each replication parts of dry material, was analyzed statistically by were uprooted at the same time and left lying on the the analysis of variance and the least significant dif ground-to die gradually. The third plot in each repli­ ferences between treatments determined. Four hours after treatment there was no appreciable cation was left as a control. Two rows of unsprnved or undisturbed plants were left as borders around each difference in the amount of inorganic phosphorus in plot. The treatments were started at 6 A.xr., and sam­ the leaves and roots of treated and check plants, but ples of 100 plants were taken from oneh replication the stems of treated plants had a significantly higher of all treatments at 4,10, 24, and 48 hr and 1 wk after amount than the checks. The uprooted plants had somewhat less inorganic P than the treated plants in treatment. The leaves, stems, and roots were separated, fresh all 3 organs. Ten hours after treatment inorganic P and dry weights obtained, and a composite sample of had dropped in all organs of all treatments except in 300 g of dry powdered tissue from each replication the stems of uprooted plants, where it was somewhat was nnalyzed for inorganic phosphorus. Aliquots of a higher; but in roots, stems, nnd leaves of treated hot water extract of the dry tissues were clarified with plants it was higher and significantly more so in the 0.5 g charcoal and used fo r inorganic phosphorus roots and stems than in the corresponding organs of check plants. The samples taken 24 h r after treatment 1 A ilm lnlalpnsl hy th e O tH re of E xperim ent S tatio n s, A erlshowed a sharp rise in the level of inorganic P in roots c u ltu ral Itew urch A dm inistration. I'SD A . N ovem ber 20, 1933 ii 5210 „ 4 ^ - 0009995" 00W2156862 IV4I9 M anuscript received F eb ru ary 11, 1033. ¿fate r * Reference--- G-Y II2S 0 4 and 1 ml of arsenomolvbdatc reagent (Nel­ son’s [5] reagent diluted with 2 parts of distilled water) are added. The tubes arc plugged with loose fitting corks and placed in a boiling water bath. A fter a 15-ruin heating period the tubes are cooled to room temperature and the contents diluted with 5 ml of dis­ tilled water. Color intensity is determined using a photoelectric colorimeter equipped with 660 mp filter. A series of tubes containing known quantities of the antibiotic are prepared and treated simultaneously with the tubes containing unknown quantities of the antibiotic. The values obtained with this series of known solution are used to calculate the constants of Beers’ law and to standardize the determinations. The sensitivity of the method varies somewhat with the particular antibiotic under consideration. The practical working range fo r Terramycin and Aureo­ mycin is from 2 to 40 pg/tube; fo r erythromycin it is 5-S0 pg/tube; and fo r carbomycin it is 10-160 pg/tube. Smaller quantities may he determined if only 1 ml of distilled water is added after the heating period, and micro cells are used to determine the opti­ cal densities. Apparently the hydrolysis with 6 N acid is necessary to obtain maximum sensitivity of the method, and use of more dilute acid resulted in re­ duced sensitivity. Only ■Terramycin will reduce the arsenomolyhdnte reagent without a preliminary hy­ drolysis, and in this instance the working range has been found to be from 20 to 160 pg/tube. Some of the data collected in analyzing aqueous solutions contain­ ing known quantities of Terramycin are summarized in Table 1. TABLE I A n a l y s i s o p S o l u t io n s t o r T e r r a j i t c m ' C o n t e n t Solution I . T n e n c , a n d M etf .h. In d . E n g . C h em . A n a l E d ., X. I3G (1020). M a n u s c rip t receiv ed J u n e IS . 1933. C olorim etric M e th o d fo r D eterm in a tio n o f A u reom ycin , C arbom ycin, E rythrom ycin, an d T erram ycin in A q u e o u s S o lu tio n D istiiicd w ater 2% Glucose 2% S ta rch D. Perlman S qu ibb In s titu te f o r M edical R esearch, N e w B ru n sw ick, N e w Jersey Wo have observed that the acid hydrolvzates of Aureomvein ( I ) , carbomycin (2), erythromycin (.?), and Terramycin ( i) react with the arsenomolyhdnte reagent to produce blue colored complexes. The opti­ cal density of the color formed has been found to he a function of the quantity of antibiotic present. Satis­ factory results have been obtained with the following procedure. Aliquots containing from 10 to 40 |ig of antibiotic are added to colorimeter tubes and the solution evapo­ rated to dryness using an air jet. Two milliliters of T erram v cin added* p g /m l T erram y cin fo u n d t p g /m l 0 10 30 100 0 10 30 0 10 30 9.7 ; 9 .9 ; 10.2 29.8; 30.7; 30.7 98.6; 99.7; 101.5 0 9.5 ; 9 .7 ; 10.0 27.7; 30.6; 30.6 0 9 .1 ; 9 .7 ; 9.7 28.8; 29.0; 29.6 0 • T o r m m r c i n h y d ro c h lo rid e w .w u sed in th c« e h tu d le a . A ll a n a ly s e s n ro p re s e n te d in te r m s of th e fre e hnse. | A n tib io tic e x tr a c te d fro m a q u e o u s s o lu tio n w ith m e th y l Iso tiu tv i k e to n e . This method cannot be applied directly to solutions containing carbohydrates and other substances which react when heated with the arsenomolyhdnte reagent. These four antibiotics may be separated from carbo­ hydrates by extraction from aqueous solution (pH 7.2) into chloroform, amylacetate, «-butanol and methylisobutyl ketone. All the antibiotic has been re­ covered in the solvent phase when equal volumes of the solvent and aqueous solution have been used. S c i e n c e , Vol. 11S 5211 000999C, I pqpqcI7Mnn » items, and leaves of treated plants; the level in the roots and leaves was significantly higher than in the checks. There was also a rise in the inorganic phos­ phorus content of stems and roots of cheek plants, hut it was less than in treated plants. The leaves of check plants showed no increase as did the leaves o£ treated plants in this phosphorus fraction, indicating that the most pronounced effect o£ 2,4-D on phos­ phorus metabolism occurred in the leaves at this time. Forty-eight hours after treatment the level of in­ organic P in^leaves, stems, and particularly roots of treated plants was significantly higher than in check plants. In the roots, stems, and leaves of uprooted plants it was about the same as in the check plants. One week after treatment inorganic P in the roots of treated plants had increased significantly and this co­ incided with a sharp decline in the leaves, indicating that it may have been translocated from the leaves to the roots. There was practically no change in the inor­ ganic phosphorus fraction in treated stems. Although by this time the level in the leaves and stems of check plants also declined, there was no corresponding in­ crease in the-check roots as there was in the treated roots. Inorganic P in leaves and stems of treated plants fluctuated in. most instances like that in the check plants, but .this fraction was consistent, and at most sampling dates, except the first, significantly higher in roots, stems, and leaves of treated plants. This experiment provides a clue to the mode of ac­ tion of 2,4-D, e.g., it may inhibit or interrupt the phosphate metabolism in the plant. These data and the fact that very small amounts of 2,4-D produce drastic effects suggest that 2,4-D may inhibit or poison the enzyme or system responsible for the hydrolysis or synthesis of the high energy phosphates. 3*1 5212 A ^ no o DOW 321(529 &• ro Vj>4 o Toxicity of 2,4-Dichlorophenoxyacetic Acid and 2,4,5-Trichlorophenoxyacetic Acid A Report on Their Acute and Chronic Toxicity in Dogs VICTOR A. DRILL, Ph.D., M.D. AND TOMIHARU HIRATZKA, M.D. DETROIT 5213 0002070 Copyright, 1953, by American Medical Association D O W 3 2 1 6 3 U Reprinted from the .1. M. A. .Ireliii-es of Industrial Hygiene aud Occupational Medicine January 1953, Vol. 7, pp. 61-67 TOXICITY OF 2,4-DICHLOROPHENOXYACETIC ACID AND 2,4,5-TRICHLOROPHENOXYACETIC ACID A Report on T h tir A cute and Chronic Toxicity in Dogs * VICTOR A DRILL. M i.D , M.D. TOMIHARU HIRATZKA, M.D. ocntoiT AMO B O T H 2,4-dichIorophenoxyacetic acid (2,4-D ) and 2,4,5-trichlorophenoxyacetic acid (2,4,5-T ) may act as plant hormones and herbicides 1 and are now being fairly widely used for the control of certain types of weeds on farm lands. They are being sprayed from airplanes on many acres of specialized farms and grazing land and are present in trademark preparations sold at retail for home gardeners. W ith the widespread use of these herbicides, possible toxic effects are important, as accidental ingestion may occur or residues may be inadvertently encountered in food. In an earlier report, 2,4-D was mentioned as being nontoxic to animals and man when administered orally .1*’11 However, the injection of large doses of 2,4-D in animals produced symptoms similar to those seen in clinical myotonia .1 More detailed observations were reported by H ill and Carlisle who studied the acute and subacute effects of 2,4-D in various species of animals.* There have been no reports concerning the toxic properties of 2,4,5-T. The present study concerns the acute oral toxicity and chronic oral toxicity of 2,4-D and 2,4,5-T in dogs. EXPERIMENTAL PROCEDURES Adult mongrel dogs of both sexes were used. Most of the animals were housed in the laboratory for a period of two to three months before the study was started. During this control period they were immunized against distemper with the Green vaccine. The dogs were fed a standard stock diet (Friskies) ad libitum. Both 2,4-D and 2,4,5-T were commercial materials, with a purity of 98.5 and 98.9% and freezing points of 132.8 and 150.6 G, respectively. In the acute studies, the calculated dose of 2,4-D or 2,4,5-T was administered as a single oral dose in capsules. The dogs were observed for a period of 14 days, at which time the surThis study was supported by a grant from the Dow Chemical Company. From the Department of Physiology and Pharmacology and the Department of Pathology, Wayne University College of Medicine. ■ 1. (a) Hildebrand, E. M . : War on Weeds, Science 103:465-468, 1946. (b) van Overbeek, J., and Velez, I .: Use of 2,4-DichIorophenoxyacetic Acid as a Selective Herbicide in the Tropics, Science 103:472-473, 1946. (r) Smith, F. G.; Hamner, G G, and Carlson, R. F .: Control of Ragweed Pollen Production with 2,4-DichIorophenoxyacetic Acid, Science 103:473-474, 1946. (. 2,4-D 14.» 93 8.8 15.2 M 12.0 14.7 11.8 174 13.0 9J 124 13.4 2.4>T 74 124 114 34 84 124 123 13.0 93 114 10.5 14.9 10.9 ' 03 ro T able 2.—Body Weight and Survival of Dogs F.ed 2,4-Dichlorophenoxyacetie Acid and Dow, Mr./Kf. a ■O redn dog the : ' 23 F........... Death Dart ! ) s 24 F........... 33 F........... s s s 37 X............ s 49 13 29 33X. 33 F.. 31 X. 35 X. s s s s 49 75 11 59 43 X. 48 F. * F In thii column menu female: X. mala. t Daily dose divided and administered one-bait In a. m. and one-balf In p. m. 1 S In this column meana survived 90 day test period. 47F. 43 X. In dogs that died while receiving 2,4,5-T, the prominent effects were weakness,' slight stiffness in the hind legs, difficulty in swallowing food, and, in one dog, bleeding from the gums. • F to tbl* t Dally dost To conserve ( d ) Blood Count: T he administration of 2,4-D or 2,4,5-T did not have any significant effect on the hemoglobin, red cell count, or total white cell count of animals that survived or died during the study. The differential blood count remained normal in the surviving animals. In three animals that died a terminal fall in the percentage of lymphocytes was observed (Table 3 ). (3 did nc and !• This : death demur early : ( e ) Organ W eights: There was no significant change in the weight of the thyroid gland, adrenal gland, heart, liver, or kidney in animals that survived the 90-day period of study. Iirtw o of the throe dogs that died during the administration 5217 0002074 s wed signs ere quiet, h in d legs, han usual hree days w ing and ;dily con1 nutcosa. of 2,4-D , there was a slight increase in heart and kidney weight. A sim ilar change w as noted in two of the four dogs th at died as a result of 2,4,5-T. ( / ) Gross P athology: T w o dogs receiving 2,4-D (26 and 34) showed areas of redness in the duodenum. W ith the adm inistration of 2,4,5-T there developed in dog 42 a diffusely reddened duodenum and jejunum , while in dog 48, autopsied on th e 59th day, a generalized icterus was dem onstrated. C « cc tc T able 3.— E ffect o f 2,4-D and 2,4J - T on H em oglobin, T otal Blood, and D ifferential C ounts Differential Count -tcid and Do* No.* Dom . U *./K *. a t ........ D eath D a jt < a r .................. ! 8 8 8 8 c ‘ i a t ........ 8 s Day H b. Om. % 0 a 90 13J0 13.82 11.70 BBC, UIL WBC C ontrol D o n 5.72 17X00 5.93 15X00 11X50 0.09 0 SO a 11X0 14.71 14.SJ 9.01 6.66 9.19 10X00 13,150 7X00 0 10 » 9 .a 10.79 12.74 9.03 5.63 ÏXZ 15X00 11X00 14.700 0 11*9 13.31 11.99 5.93 5.78 9.47 14X00 14,900 9 ,4 a 17 M................. a 90 s B as., % E osId ., % D eath Day a a 27 0 0 1 0 0 0 6 7 0 •• n 11 ii 79 a 9 7 0 0 0 0 0 1 0 a. 67 56 28 1« 18 1 1 1 0 0 0 « 1 4 1 0 0 0 4 7 4« 39 a 51 61 66 17 a 5 4 4 « i 0 0 0 0 66 61 n a n 55 a 1 0 0 ,, „ - 14-D 49 18 S U ................. at zs 0 10 8 1 . ....... 8 8 8 8 .g 8 8 8 at 90 0 a 49 MU........ ISM................. a 11X00 10.41 11.44 n_a 5.01 606 (J 5 15,400 m a TO a 11.19 14.49 14J9 5.5« 5X9 5.67 14X50 9 ,7 a 15X00 54 47 79 41 14 4 7 tt 4 0 lia m is x a a 44 4 9 14.90 147 i 1M 79 a 11 14X9 11,150 9X S 17 4 U U « a 9 0 0 0 1 9 •0 1 •• 49 19 0 0 12 0 0 0 10 0 0 « •• 79 i a 1.4X-T 49 U........ 0 11 59 49 T _ ....... 1 n 15,450 «X a 77 91 7.10 19X50 11X00 a U 0 1149 14.M IS 4 47 r. ........ 0 12.44 in nxa a a a a 49 U................. 0 1149 U S 11.97 9X1 i«xa in 14X50 7X79 n n 44 19 a a ~~ 13,4a 49 a - 1194 194 IS 199 1X7 . a ■eakness, )ne dog. ' N eut.. Lym ph., M on., % % % 114 n M 9 9 1 1 0 4 4 i 7 0 •• a 0 1 1 0 1 m m 0 0 0 0 a 11 *1* to th li eolum a maana fem ale: U . m ale. t Dally dose divided u d edralnlstertd one-belf In *. m. end one-belt la p. m. To comerve iptce, only date on control dots end tbe hl*hest dole* a n (Iren. have any count of rem ained ill in the it of the .ived the listration ( g ) M icroscopic C hanges: T he heart, lungs, thyroid, adrenal, ovary, o r testes did not dem onstrate any significant changes. T hree of the dogs receiving 2,4,5-T an d tw o receiving 2,4-D showed an occasional area of focal necrosis in the liver. T h is finding, however, was not related to either the dose of the com pounds or the death of any animal (T able 4 ) and was judged to be of no significance. T h e duo­ denum of dogs 26, 34, 42, and 48 showed varying degrees of hyperem ia and some early infiltration of cells in the mucosa. T he subinucosa and serosa were norm al. 0002075 5218 ai oc 6 A slight increase in the number of casts in kidney sections was noted in some dogs (Table 4 ) , but again this was unrelated to dosage and was of doubtful significance. diffe COMMENT In the acute studies, single large oral doses of 2,4-D administered to dogs may produce signs characteristic of clinical myotonia. This effect of 2,4-D was similar to that observed after various experimental species had received injections.1** In addition, some dogs showed signs of irritation and pain when the skin of the back of the neck was grasped, which may indicate meningeal irritation. The extension and crossing of the hind limbs, when lifted by the shoulders, may point to a spinal of a O Z ZP to T able 4.— H istological Changes in L iver and K idney o f D ogs F ed 2,4-D or 2 ,4 J -T fo r 90 D ays Ci CO C” Llvtr Doc No. D o«. M f./K c. 23............................. 24............................. 25............................. 26............................. 27............................. 26............................. » ............................. 30............................. 31............................. 31............................. 33............................. 35............................. Son» Son» t 1 5 5 10 10 10 so 20 so 36............................. 37............................. 38............................. 39............................. 40............................. 41............ .......... 42............................. 43............................. 44............................. 45............................. 46............................. 48................... ......... Non* Non* S s Death D ar 49 ss Conce* tlon S.4-D 0 0 0 0 1+ *+ 0 0 1+ 0 1+ s+ Focal Xecroit* Ca*t* T ab. Deem . 0 0 0 0 1+ 0 0 0 0 0 1+ 0 0 0 0 1+ 0 1+ 0 0 0 0 0 0 0 0 0 0 0 1+ 0 0 0 0 0 1+* 0 0 0 1+ 1+ • 0 0 0 1+ 0 0 *+ 1+ 0 0 0 1+ 0 0 . M s s 10 10 10 20 so so .. 49 7# so 0 0 0 0 0 0 Î+ 0 1+ 1+ 1+ *+ E cant of th 2,4,5 perip leucc dogs days, perip L occas chror. signii 2,4,5organ 0 0 0 0 0 S.4.4-T „ tratii ness, All f and sliglr dose WitI t: appre produ oral L mg. p spastic 0 0 1+ 0 0 0 0 A! 2,4,5-' sympt 0 * Al*o a l + fibroil*. T Iuuc* e n d e d on bast* o t 0, 1 + , S + , t+, and *+ . I central lesion. Other' effects occasionally observed were sneezing, rubbing of the eyes, and diarrhea. The effects of 2,4,5-T, previously studied, were not as severe, being limited to stiffness in the hind legs and ataxia. The higher doses of either compound produced anorexia and weight loss. Although initial effects may be seen six hours after the compounds are given, the syndrome developed slowly in the dogs, and death was delayed, occurring several days after the administration of the compounds. In the chronic studies, the dogs survived doses of 2, 5, or 10 mg. of 2,4-D or 2,4,5-T per kilogram of body weight without apparent ill effects. However, the daily oral administration of 20 m g./kg. of 2,4-D produced death in three of four dogs within 18 to 49 days. This delayed death may indicate a cumulative effect of 2,4-D . The signs observed in animals dying from the 20 m g./kg. dose of 2,4-D ♦ T1 died, ; study. gums, swallo in the De to patl 5219 0002076 f I 7 some dogs ignificance. i I fo r 90 D ays lu tj ------------ —> Tub. Dcs»n. 0 0 0 0 0 1+ 0 0 < 0 0 0 SUMHABY s 0 0 0 0 1+ 0 0 0 0 0 0 t ng of the as severe, s of either ay be seen ivly in the tion of the f 2,4-D or wever, the ce of four e effect of : of 2,4-D i The acute oral L.D.M of 2,4-dichlorophenoxyacetic acid (2,4-D) in dogs was approximately 100 mg. per kilogram of body weight. Doses in this range or higher produced a definite myotonia accompanied by anorexia and weight loss. The acute oral L.D.so for 2,4,5-trichlorophenoxyacetic acid (2,4,5-T) was in the range of 100 mg. per kilogram or higher. Such toxic doses produced only signs of a mild spasticity. All dogs survived the oral feeding of 2, 5, or 10 mg. per kilogram of 2,4-D or 2.4.5- T 5 days a week for a period of 90 days. These doses did not produce any symptoms or changes in body weight, organ weights, or blood count. Three of the four dogs receiving repeated doses of 20 mg. per kilogram of 2,4-D died, and all four dogs receiving 20 mg. per kilogram of 2,4,5-T died during the study. Toxicity to 2,4-D at the dosage level was accompanied by bleeding from the gums, necrotic changes in the buccal mucosa, and some difficulty in chewing and swallowing, but there was only little evidence of clinical myotonia. A terminal fall in the percentage of lymphocytes was observed in three of the animals. Death during the repeated administration of 2,4-D or 2,4,5-T was not related to pathological changes in the liver, kidney, or other organs examined. Printed and Published in the United States of America 0002077 UOW 321636 dogs may ras similar ions.1’* In the back of ension and 1 spinal or differed somewhat from those seen in the acute studies. With the chronic adminis­ tration, the animals exhibited chiefly a stiffness of the hind legs and ataxia, weak­ ness, difficulty in chewing or swallowing, and occasionally bleeding from the gums. All four dogs which received 20 mg./kg. of 2,4,5-T succumbed between the 11th and the 75th day of the study. Toxic effects in these animals consistently included slight muscle spasm and difficulty in swallowing food. Thus, the chronic oral toxic dose for 2,4,5-T in dogs seems to be in the same range'as that obtained with 2,4-D. With both drugs, there was a terminal loss in body weight. Dogs surviving the chronic doses of 2,4-D or 2,4,5-T did not show any signifi­ cant change in hemoglobin, total blood counts, or differential blood count. In three of the dogs (64. 57, and 105) which died while receiving 20 mg./kg. of 2,4-D or 2.4.5- T, there was a definite decrease in the percentage of lymphocytes in the peripheral blood. Hill and Carlisle * have observed a fall in the polymorphonuclear leucocyte count in one dog and a decrease in the per cent of lymphocytes in two dogs to which 50 mg./kg. of 2,4-D was administered intravenously each day for six days. In mice which received chronic injections there was no alteration in the peripheral blood picture.1 Large doses of 2,4-D administered intravenously or, in the present study, orally occasionally produced some necrosis in the liver. However, the administration of chronic oral doses produced only a slight and inconsistent focal necrosis of doubtful significance (Table 4). Deaths during the chronic oral administration of 2,4-D or 2.4.5-T were not correlated with significant lesions in the liver, kidney, or other organs examined. 5221 K * n * SUSPECTED POISONING O F DOGS FROM EATING GRASSES TREATED WITH 2,4-D D. L.-Baker, O.Y.M., F. K. Ramsay, D.Y.M., M.A., and E. P. Sylvester, B.S., M.S., Ph.O. Iowa State College Ames, Iowa I •4' V. ’/ • * i&■ ■ - , v . ; ‘ I-? ■ f. ,«*'y Frequently dogs are admitted to the vet­ erinary clinic with the history th a t the owner feels th at the animal has been poisoned from the effects of having con­ sumed grass treated with 2,4-D. I t has been commonly observed th at dogs, on oc­ casion, will eat various quantities of grass. We were unable to find any information in the literature concerning field cases of the toxicity of 2,4-D-treated grasses in dogs. Hill and Carlisle1 have noted that, by intravenous daily administration of from 25 to 200 mg. of 2,4-D per Kg. of body weight, effects ranging from sub­ acute intoxications to death may be pro­ duced. They further noted that dogs in­ jected with 2,4-D showed a considerable susceptibility to the development of liver damage. Other work concerning 2,4-D poisoning in various animals has been done by other workers.3'* Since there is so little information available on 2,4-D poisoning in dogs, in which the 2,4-D was consumed on treated grasses or administered per os, the following trials were conducted. Individual, heavy stands of lush grow­ ing bluegrass, orchard grass, timothy, and quack grass were sprayed w ith the water emulsion ester (butyl ester 4 Ib./gal.) form of 2,4-D, a t the rate of 4 pounds per acre. These are the most common grasses that dogs are most likely to eat. The spraying was a t least twice as heavy as ordinarily recommended and sufficient to cause foliar damage to the grasses. The spraying was done on June 18, 1952, and the grasses were harvested in four sepa­ rate lots (112 Gm. each) 48 hours later, on June 20, 1952. These grasses were chopped in small pieces and mixed in pro' Dr. Baker Is aaociato protestor of veterinary med­ icine. Dr. Bamsey is associate' professor of veterinary pathology in the Division of Veterinary Medicine and Dr. Sylvester is extension professor of botany and plant pathology194 portio S S R W y j y w ith fresh horsemeat and dog meal. This mixture was fed to two healthy one-year-old mon­ grel dogs, weighing 20 to 25 pounds each, in .equal .amounts, in three consecutive feedings, tw o on June 20 a t morning and evening meals, and the last one a t the morning meal on June 21. Apparently the palatnfoHity of the food was not altered appreciably, as the dogs immediately con­ sumed all of the food. The dogs were observed but no change in attitude or signs of any ill effects could be detected a t any time within 96 hours following the first meal. On June 24, both animals appeared to be in excellent health, and a subsequent trial was instigated. Each animal was given 5 cc. of the same ester concentrate of 2,4-D orally in a gelatin capsule. This dosage represents approximately 500 mg. of 2,4-D per Kg. I t is noteworthy that this was a single dose given per os. No clinical deleterious effects were observed a t any time within 96 hours following this oral administration. On June 28, one dog was sacrificed. Necropsy examination of this animal failed to reveal any macroscopic lesions. The other animal, under daily observation, remained in apparently excellent health for the fol­ lowing 82 days, a t which time these experi­ ments were considered concluded. We have not arrived a t any definite con­ clusions with this meager information. However, in most instances of suspected 2,4-D poisoning entered a t th is clinic, some other cause has been definitely estab­ lished. We are not cognizant of a proved field case of 2,4-D poisoning in dogs having occurred in this area. • i/ t lV [fas2 Clini Sodi 400 H. W . Monroe, i F. E. Ec! Detroit, l e [ 4 i ) Intrav tended c recent : been eir agents anesthes: the oldei every br inductior cedures. The a constant! thetic in m et the anestheti proaches paper is in dogs i To ap anestheti low toxic cated, an side reac desirable investiga clinical ii Material i References Since J administc hospital, action, dc period of sodium w the very about equ was used 1. Hm. E. V- and CaxUalo. H.: Ind. Hyg. & Toxicol, s e (1947) 33-93. 2. Ennis. W. B.; Thompson. H. S - and Smith. H. B.: Tributyi phosphate as a solvent for pnparins concentrated and oil solutions of 2.4-D acid and similar substances. Science te e (1946) 473. 3. Grigsby. B. B . and Curtson. B. 1\: Soma effects of herbicides on pasture and grazing livestock. Mich. Agr. Exper. Sta. Quar. Bun. aa (1850) 378. 4. Bildebrand. S. M.: War on creeds. Science te e (19«) 463. 3. Bildebrand. S. H.: HerhlcMal action of 2.4-D on the erater hyacinth (Xichontte craasipes) Science 103 (1943) 477. 6. Smith. T. O.i Hammer. C. I*, and Carisnn. B. 7.: Control of ragweed- pollen- with 2.4-D. Sdaswo te a (19«) 473. Dr. Knlrk stofi of th Parke. Davi. •Thiâmylâ March. 1953 The North Americas Veteriaeries JIW JJ 0012187 5222 5223 Ä i C H E M IC A LS A RE E F F E C T IV E /or Woody Plant Control in the South By A. H. W a l k e r , Extension Range Specialist, Texas A and M College System, College Station, Texas all states but neither is die brush problem so immense. Texas has some 63 million acres infested. Sometimes the brush infestation is so heavy that live­ stock h an d lin g on the range is difficult. Often this brush grows on the most productive land. Some brush con­ trolled areas have produced five times as much grass as untreated ones the year following treatment even under drouth conditions. Average production increase following woody plant control usually varies from 20 to 50 per cent, depending upon rainfall, site and management practices. This means more pounds of meat per acre and increased efficiency o f production. In timber areas, hardwoods often hold back or crowd out desirable pine trees. Control of these hardwoods increases the quantity and quality of for age production and also makes livestock handling easier In forest areas the release of desirable timber growth i: both practical and profitable. Proper management following brush control is ven important. Control of woody plants so that an operator car continue to overgraze is folly. Brush control is just ont phase of a pasture management program just as building a tank or constructing a fence. For permanent benefits tc result to a pasture, certain range management practice such as proper stocking, deferred or rotation grazing ant in some cases reseeding and fertilization must be followec W e are primarily concerned with chemical control c woody plants here and it does offer the most promise of an one method. Mechanical equipment such as bulldozer: anchor chains, brush and weed cutters and root plows ofte: have a place in brush control and maintenance work. Some times a combination of mechanical and chemical method, w ill give the best control per dollar spent. Controlled burr, ing in timber areas to increase grazing can be utilized L some cases. N o one method of brush control can be recoir Editor's N o te : The use of chemicals to increase forage and livestock produc­ tion by the control of undesirable brush on range land and pas­ tures is relatively new and challenging field. The author is among the first to conduct an extension program which has brought such information to ranchers. a W oody plant contpal-is-one of the biggest problems faced by the farmer, ranchman and timber producer in the South today. There are about 240 million acres in the United States infested with undesirable brush and nearly half of this, or 108 million acres are in the fourteen southern states. W ith this brush under control, how much would it add to livestock producers’ income?— possibly one-third of a billion dollars annually. Undesirable trees and brush use up water and minerals which should be available for forage growth for increased livestock production. Some water resources information recently compiled by the Experiment Station in Texas illustrates this point. These data show that the annual average precipitation amounts to 361 million acre feet of which 131 m illion acre feet is lost to brush and weed growth, 44 m illion acre feet runs-off and only 30 million acre feet is used by desirable plant growth of all kinds. Actually, brush and weeds use one third o f all precipitation received. W e hear a lot about un-used run-off and find in Texas that three times as much water is utilized by worthless brush and weeds as runs off down our rivers. Actually, brush and weeds use four times as much water as all desirable plant growth— this includes all cultivated crops as well as range and pas­ ture grasses. W e have a potential of four times the pres­ ent production in Texas if this water could be properly utilized. Lack of adequate water is not such a problem in Top Left: Pott oak and blackjack oak aerially sprayed in May 1952, shows 63 percent root kill 18 months following treatment. For­ age production has been materially increased. (Texas Agri. Expt. Sta. photo). Lower Left: Cut-off stumps treated with 2.4J-T in diesel oil did not sprout. Good native grass made a quick come-back. Right: The author examines the trunks of post oak trees basal sprayed with 2,4J-T in oil two years be­ fore. On six inch trees and larger, frill treatment is cheaper and is very effective. 10 5224 M 2,4,5-T ds to •¡vi.makiii*“' JM 'OKM ATIOX OX I I e UUICIUES AND A S KVAI.UATIOS OK T llK IB U s e receiving ¡,4/ hT were slight mustele spasm, and difficulty in swallowing. llisrnlogicnl studies re* vealed mild change* in the liver and kidney*. The A nt experimental toxieolocieal work eondneted on farm animal* wan that of Mitritoli and TABLE 1— Mato rial* Studied* Csmmon a a m a 3 .4 - D 3 .4 ,5 - T • M CP S - M s th rl- 4 - e h la ra p h ra a s ra c M i« a d d 4 - C h la (« te to la x )ra < * < le a d d 3 - ( 3 .4 .3 - T ric h te re p h a n a s r ) - p ia p to n ic a c id Silvas * lh « M la tte a « . C h r m ic s l n a m e 3 ,4 - D ic h la ra p h m « x ra c r(i< a d d 3 ,4 .3 - T rt c h la r o p k m a x jr a c X l* a d d a n W ife s h a m ic a l* « M i ia h a rb id d a l fo n a i- eo-workcr».* Thews investigator« pastured sheep and eons on folia go sprayed with more than rcrommended amounts of 2,4-D without effect. Thoy also fed a loetating cow SJi Gm. of 2,4-D daily for 10(1 days without ill effects. Milk from the cow did not contain a demonstrable quantity of 2.4*1). No effects were observed in the ealf to which thia milk wan fed. nor was there eny 2,4-1) found in the serum of the ealf. The serum of the cow, however, contained 8.4 p.pjn. of 2.4-D though none waa found in the liver, kidney, or fatty tissues. Early ia IPSO, flrigshy and Farwell* published the results of experiments with 2,4-D and 2,4,5-T involving domestic livestock. In thia study, alfalfa w ai sprayed with various herbieidal formulations including three different 2,4-D preparations and on* 2.4,3-T preparation. Livestock including horse*, daily and beef cattle, sheep, swine, and chickens were immediately pastured ia the freehly treated areas. In summary of thia work, the authors stated, “ Result! of thia experiment indicate th at none of the herbicide* used had any serious physio­ logical effect upon the livestock involved. They also indicate th a t none of the livestock preferred any of the sprayed arena to those th at were unsprayed; however, with the lots sprayed with 2,4-D herbicide«, the livestock grazed the areas sprayed almost as woll aa the nasprayed. 8inec the rates used were two to four times greater than recommended dotage, it seems th a t the farm use of these materials for pasture weed control is a rea­ sonably safe procedure." Mattsiala S t u m d Commercial herbicides generally ran lie described as formulations of active ingredients. The active ingredient usually appears ns a salt which ia solu­ ble ia water or as an ester which ia soluble in oiL Formulations may lie designed for use as water solutions, w ater emulsions, oil solutions, oil emul­ sions with water, or dusts, depending upon the use for which they are intended. Basically, most fo r­ mulations contain, besides the active iagrrdieata. a dispersant, a solvent, a wetting agent, and per­ ils ps a diluent. These a n usually considered to be inort ingredients although thoy may have a pro­ found inducnes upon herbieidal effectiveness. Ia the studies reported herein, toxieologirai d ata a n reported not only fo r the basic aetivc ingre­ dients aad their ehemieal derivatives (table 1) bnt also for the Aniihed formulatioas as they are marketed* (table 2 ). Oual A D M unsnunost o r S u r a u Domes Procedure.—The toxicity of the various herbi­ eidal materials aad formulations wfaea adminis­ tered ia single oral doses to various s pee ics of animals baa been determined. All rata, guinea pigs, and rabbits need were young adult animals from the stoek 'colonies of this laboratory. The mien w e n young adults obtained from Garworth Farms. The ehieka used were New Hampshire Beds obtained from a commercial hatchery when about 3 days of ago aad were about 3 weeks of age when treated. The teat awterials were administered by iatulwtion unless otherwise indicated. Aqueous solutions were used wherever possible. Olive oil or corn oil solutions w en used where w ater was not appro­ priate. Surviving animals were observed until re­ covery waa certain, usually about two weeks. Remit».—The resulta of the acute oral studies are summarized in tibie* 3 and 4. • I t s U a d e -sa sM d d ra g a lis te d ia ta b le 3 s a d i a th e te s t a n « re d a c ts e t T h e D e w C h e m ic a l ( V , M id la n d . K ic k . TABLE 3—H erbieidal Form ulations Studied T n d sa ia s Aativa iaeredienss ( » ) 4 5 .0 A lk a a a la m ia a s a lte W 1 .4 - 0 3 .4 - D e w w « * d k ille r (fo rm a la 4 0 ) 44.0 Imarapjrl etter al 3,4-D E s t e r a * 3 4 S (e ld f e r a n ls t ia a ) E s ter e « 3 4 5 ( pr w s t f o r m a l* lia s ) E s te re « te n -te a B r a s h k ille r S O -M • B r a s h k ilt e r T E s t e r a « 7 « (u a s d ia * 0 s n ia tia a a a ljr ) E s t e r n a 7 S E (a s a d i s s ilh a r *a a s ia t ic a a r w a te r s m a ls ia a ) E s t s r o a b ra s h k ille r (a id fo rm « 1 s t t e a ) E s t e rn a b ra s h k ilt e r (p ir n »a t fo n o « la i t e a ) E a r n s (w a s caU ad H - 1 0 7 1 ) D a w M C F s m in a w a a d k ille r (J ^ J 3 3 .3 T s a p ra p y l « s ta r o t 3 .4 ,5 - T ; 1 3 .1 M lz c d a m p i r a t r r s a l 3 ,4 .5 - T « 5 .3 M a n te , d i- , u ip r a p iim s (b r a s i h a t e i r t h a r « a te ra a l 3 .4 ,5 - T 7 0 .3 M a n te , tr ip ra p jr la a s ( ly e s l b e te l a th a r a tte ra a l 3 ,4 - D 3 7 2 Matei « s t e n a l 3 .4 - D : I l i Sauri s a u ra a l 3 ,4 > T SM Batyt estere al 3, 43-T 3 7 .1 Im p r o p r i s s ta ra o l 3 ,4 - D ; < 3 t .4 a - B a t e i « s ta ra a l 3 ,4 - D M J Is a p r a p il « ste r i a 1 3 ,4 - D ; M J B s t y i s t e s i* « 1 3 .4 - lb 3 5 .5 la s p ta p irl « s t e n a l 3 ,4 - D ; 3 4 .4 Ia a p ra p y t a m s rs a l 3 .4 ,5 - T 3 4 J M a n te , d i- , tr ip ro p jrls a a ( t e s a i b a te t « th a r m ta rs a l 3 ,4 - D i 3 3 .0 M a n te , 41-, tr ip r a p r ia a a (t e c a i b a te t c ib a r m u r a « 1 3 ,4 .5 - T 6 4 .5 M a n te , di-, tr lp ro p rt e a * ( ir n a l h o te l U b a r ta ta ra a l *11v n 5 5 .1 A lk a a a la a u n a s a lto s i M C P 1 .5225 li V ‘00287.0 624 Y. lv. Kowe ano T. A. II vjia» In nil cases where the (lata were suffi­ ciently extensive to allow statistical analy­ sis, the l.d.3, values with their 19/20 confi­ dence limits were determined by the method of Litchfield and Wilcoxon.T In Am. J. V«t. Ko . U fTtim 1044 all other eases, the Ld.*, values were esti­ mated and the range between the dose al­ lowing survival of all animals treated and the dose causing all aiiimulx to die is given. F o r comparative purposes, some of the vai- TABLE 3— Sum m ary of A cute O ral T oxicity of V arious Basio Horbicldal M ataríais • Ldue • (19/30 ennddaaca Iboita) Material Btx Vrhirie SpfHn (mt/kt-l 3,4-0 (3,4-Dlchlarnphtnoxracaiic acid) Rat« 375 (303-404) M Olive eil Mice MR (313-434) M Oliv»eil Guiñe* pift MaadP Oliveeil 440 (397-443) Chicha MeodP Olivaeil Mi (354-417) Rane« Do*« (4) Captata 100 (35-350) . ■ * Rane* 3.4-D, »Ihttaltaiint aalta — (340-704) Chiche (1) Water (dea (eoa . ' •• : ■ . ’ add equlvaient beala) 3,4-D, n4ha Mit tos (410-1.003) Beta T Water iT— I- 440 Reta (3) Water . Ceinro pipe 4SI (417-737) M Water ---, anise* pipe (3) Water 1.000 --374 Mire (3) Water 400 -Rabbita (3) Water 3,4-0, ieepreprt atar Reta Mend P Oliveeil 700 (549-401) 440 (441-471) Guión pia M Olivedi 441 (304-744) . Mice M Oliveta 1,430 (1.137-1.749) Chiche MaadP Olivaoil 3,4-D. a M betri «ton 430 (330-944) Reta F Carota 444 (404-1.IM) Oaíete pife T CeroeQ ¡>Rabbita F Cernea 434 (353-713) Ranre Mire 713 (400-1,040) F Cernea MendP Undiluted 3,000 115904,940) Cblrke 3,4-D. »cae-. 41-, tripreyrleae (treat botri 170 (410-4401 Rué P Cunta «thertetare 3,4,5-T (3.4,5-Trichlonpheoesraeetie uU) Rete 100 (301-440) M OliveeO Mice M Oliveta *49 (345-419) . 341 (107-473) Quinte pin MendP Oliaceli MaadP OliveeO >10 (311-454) Chiche Raute 100 (40-340) De*e(4) CapeeIt 3,4,5-T, ieepreprt eater 494 (430-594) (Rata MendP OUvoeU P OliveeU 440 (343-447) /Ouiueo pige P OliveeO MI (190-799) .Mice 3.4.5-T, auxed botri aten Ret • f Cernea a i (919-739) Rente Rabbit 713 (5OO-1.000) M Cenali 9 40 F CaneO (•74-1,313) Mice Rente P CeneO t u (404-1,000) Scinco pige • Rente 3,4,S-T, alni Miri teten 740 (400-1.000) P OUveett Rota Reefe MCP (4-eklere-e-tatoxraeaticaciderS-Metkri700 (400-1,000) Rot M Cenca 4-cklsrephtnareeetia odd) Reefe MCP, etaiooioU Rat M Water LSOO (1,000-3,000) Kenia L300 (430-1,000) Onion pife M Water caves, (3(S.4.3-triehleropheaeBy) pnpieote 4 4 0 (500-740) Kat MaadP ContU Rente •aid) •flea, wived botri eetan ~Bat 900 (350-1.000) 7 Canafl Rente Rabbita 750 (S0O-1.00O) P Undiluted Chicha UM (707-3.0M) MandP Con til (Ovos. Bese-, di-, trlproprleoc fijad botri 431 (473-414) Rot P CeneO ether atan Ranen OeioMpifa K CeneO * 1,340 (500-3.000) Rente Mica P Con eil 1,410 (1.000-3.0M) Chicha LIM (447-1,470) MaadP Candi Rabbit« 410 (410-1.070) P Undiluted o O , 508765 5228 lisi.J.V kt.I(k». IX K O IC M A T IO N O S O c n u i i 1(1.14 Il£ U IIIC II> E S AN O A N K V A J .P A T IO X OK T l I K J K U « K n<*s givcii in flic lite r a tu re a rc included in ta b le 3. Toxic symptoms generally observed in animals made ill with this type of com* pound include loss of appetite, loss of weight, depression, roughness of coat, gen­ eral tenseness, and muscular weakness par­ ticularly of the posterior -uarters. Post­ mortem findings usually include irritation of the stomach of small animals and of the abomasum of rum inants, minor evidence of liver and kidney injury, and in some in­ stances congestion of the lungs. Ooal AminnsTXATioN* o r S n u n s Dosxs o r 2.4- 0 t o B a t s a n s C h ic k s Experiment 1, S a lt. — Procedure. — M a tc h e d groups of 5 or 0 ynung adult female rata from tho stock colony of this laboratory wore fed 2,4-D five times s week for four weeks by intubation. The dosages employed wero 0.0 (controls), 3.0, 10.0, 30.0, 100.0, and 300.0 mg./kg. adiniuisterod as aliquots of olivo oil solutions emulsified in about 2 mL of 3 to 10 per cent aqueous gum srabic solu­ tion. The controls received appropriate doses of olive oiL Result*.—The control animals and those animals receiving 3.0, 10.0, and 30.0 m g./ kg. of 2,4-1) showed no adverse effects as judged by gross appearance and behavior, m ortality, growth, hematological values, blood urea-nitrogen concentrations, organ weights, and gross and microscopic exami­ nation of the tissues. Hats receiving 100.0 m g./kg. showed varying degrees of gastrointestinal irrita­ tion, slight cloudy swelling in the liver, and a depressed growth rate. Those receiving 300.0 mg./kg. failed rapidly and died. Se­ vere gastrointestinal irritation was die principal adverse efFcct observed. Experiment 3, B u t i.P r o c e d u r e . — M a tc h e d groups of 3 young adult female rata from the stock colonies of this laboratory were placed on diets containing 0 (control), 100, 300, 1,000, 3,000, and 10.000 p.pju. (parts per million by weight) of 2.4- D. Results.—The controls and those animals receiving the diets containing 100 and 300 p.p.m. of 2,4-1) fo r 113 days showed no ad­ vene effects as judged by food consump­ tion, growth, general appearance, m ortal­ ity, blood nrea-uitrogen concentrations, he­ matological examinations, organ weights, and gross and microscopic examination of the tissues. Those on the diet containing 1,000 p.pjn. for 113 days suffered slight adverse effects fi-».» characterized by depress«! growl It rale, excessive mortality, slightly increased liver weights, and slight cloudy swelling of the liver. Those on the two higher diets were de­ stroyed after twelve days as they were not eating aud were rapidi)' lwiug weight. Ex­ amination revealed increased liver and kidney weights and slight pathological changes in these organa. Experiment 3, Chick*.— Procedure.—Twelve New Hampshire Bed chicks weighing from 210 to 340 Om. each were divided into four groups of 3 chicks each and maintained for aerea days on diets containing 0 (controls), 300, 1,000, and 3,000 p-pjn. of 2,4-D. Results.—The only adverse effects ap p ar­ ent grossly were a redaction in food intake and a retarded growth rate in the birds fed the diet containing 3,000 p.p.m. of 2,4-D. No histopathological examinations were made. Osa i. A sso n sn u io M to S t u m Purpom.—Since essentially all of the toxicolog­ ical information oa 2,4-D aud 2,4,3-T type herbi­ cides was obtained using small animals and since the practical application of these m aterials in-, voIves exposure also to large aninuiis, experiments wero undertakes to ascertain if extrapolation o f the d ata obtained oa the laboratory animale to largo animals was justified. Estero a Ernsk Killer.—Sineo e s t e r o a b r a s h killer (formulation of esters of 2,4-D ami 2,4,3-T) is perhaps the widest used of tho products dis­ cussed herein, it was ehosea for study. In tho experiments outlined in tho following paragraphs, administrations wero all by intuba­ tion. Treated animals wero constantly under ob­ servation and tho ordinary clinical olwerrations were routinely made. In general, only punitive observations are reported. Experim ent 1.—A steer weighing 291 kg. was given a single done of 1,000 m g./kg. No perceptible symptoms of toxicity were observed. Experim ent 2.—A steer weighing 293 kg. was given 1,000 mg./kg. of estenui brush killer on each of three successive days. General depression, decreased food and wa­ ter intake, and decreased rumen m otility .were observed following the th ird dose. These symptoms became increasingly se­ vere until the steer died on the tin n ì day following the lost dose, i Death was undrainatie; no convulsions, struggling, or signs of pain were observed a t any time. Nec­ ropsy revealed the following: Bumcn con­ tents were dry and .-:..ellcd strongly of the 5227 62« AM. J. V*KT. UIS. H W « » 134« V. K. Howe axd T. A. II ymas examination of tissues from the lung, heart, spleen, adrenuls. pancreas, thyroid, thymus, bladder, and lymph nodes revealed no ab­ normalities. Sections from the liver re­ vealed small areas of focal hemorrhagic necrosis surrounded by areas of fatty de­ generation. In the kidneys, very slight in­ terstitial edema and congestion in the me­ dulla and cortieal medullary region were observed. Although gross examination of the gastrointestinal tract revealed mild irritatiou in the duodenum and abomasum, sections of these organs failed to indicate any changes of significance. Kuron ( II -1078).—Because of the cur­ rent interest in derivatives of chlorinated plienoxypropiouie acid, one of the more promising formulations, kuron (table 1). was included in the work with eattie. A steer weighing 230 kg. was given 100 m g./ kg. of kuron on each of fifteen consecutive days. No adverse effects were noted when judged by general appearance, behavior, and weight gain. The animal was killed fo r examination forty-eight hours following the last dose. Gross examination of the in- TABLE 4— Sum m ary of Aeuto Oral T oxicity of V arious HcrbicM al F orm ulations (Dow) Valeria! ■ Spati*« Max 3.4-Umr m i kfllar Colata pia« f Water 3.4*13*« c H kinrr (fa m u li 40) Bau F Water Velici* Satana 44 Hata X Olir*«) E r t a la * 3 4 S (a U ) Bau r Esulala* io «ater Sam oa S4S ( m v ) Sam oa i*a*Ua Bniafe killer SO-SO Baia Rat« RaU Cuiaaa pica Mica X lilla* all (llia* eli Rauiaiaa io «ater KaniUian ia «alar Cera ai) Kraal kUlarT Rabbiu Ckicka Rat Calata pio* S in Rabbiu Ckicka r F T T r Undllntrd S a n a r Undiluird F KanUino In «nter y Kmuiaiaa In votar y mire ail X Undiluted U nod K Undiluted Hruak kBarTO KaU — r C an oil Bra«h km»* TOE Rat« K rnrn nil Salam i Woafc kUar (a U ) Rat» Rata (•aiata pia» Coinça pia» Rabbiu X Kmulnim in «nier ■31 end F V a n oil X Corn nil t F l ’ora nil V and T Cara oil Click» SUar» X and F Tarn ail CadilMtd S a u n a k n u k k iU e r (s a w ) lAee (10/3U ronSdeae* liait«) (a c J k c .) 3.MO <1.000-3.000) Rane* MO (700*1,000) Raaao •SO (300*1.000) Kaaaa I.D O ft (300-3.000) Rana* 4041 (600*1.000) TOO (660-600) I.O TO (700*1,060) 1.160 («30*1,630) »00 (634*1.070) Raaa* 1.430 (600*3.000) 4.0011 (3.700*6.900) 1 .2 0 0 (763*1.660) 1.410 (676*3.300) 1.230 (036*1.630) « 4 9 (604-1,190) s,mN) (1.330*3.940) Itaaa* 730 (M O .I.IM 0 ) Kara* 300 (3 6 0 -I.IH H I) Rs«ire 1.000 (300-3,000) 4 M («00.960) I.3 3 U (1,040*1.430) 1.400 (1.390*1.640) MO (7 9 0 -1 .1 6 0 ) Rana* 3,000 (1.000-3.000) Graaur than 1,000 5228 0002323- OUW508767. cstcruu: tile uboinastuu was impacted and the coni cuts of the intestines were entirely fluid: the mesenteric vessels were congested and the spleen was dark and shrunken. Experiment 3.—A steer weighing 295 kg. wax given 300 mg./kg. on each of two con* scent ive (lavs. On the third day, the animal was off feed and nnueu motility had essentially ceased. On the fourth day, the steer appeared perfectly normal and there were no discernible after-effects. Experim ent 4.—Another steer weighing 336 kg. was given 500 m g./kg. on each of three successive days. No toxic symptoms were observerd and the animal remained on full feed. Experim ent 5.—A steer weighing 230 kg. was given 100 mg./kg. of esteron brush killer on each of fifteen consecutive days without any outward appearance of ad­ verse effects. The animal was killed for study forty-eight hours following the last dose. Gross examination of the internal or­ gana revealed only slight petechial hemor­ rhage in the. duodenum and a mild diifuse irritation in the abomasum. Ilistological «27 I xwmwiatiox ox Iltam ciuts axd ax E vall-atiox ok T a g » Use s< stm t»! the similarity in susceptibility of cattle and small laboratory animal* to this type of herbicidal materials. I t has also been demonstrated that cattle are distinctly more tolerant of this type m aterial than dogs. The following conclusions with regard to cattle seem justified: 1) A daily dose of 30 m g ./k g probably can be tolerated for prolonged periods without adverse effect. 2) A daily dose of 100 m g./kg. would D iscussion' not be expected to cause any ill effects un­ Siyitificauce of Experim ental Result*.— less exposures were continued fo r a week or Study of the data presented in table 3 in­ longer. However, if exposures were con­ dicate* that the various forms of 2,4-1) and tinued, some liver and kidney injury, aud 2,4,5-T all fall in the same range of toxicity perhaps some gastrointestinal irritation for rata, mice, guinea pigs, and rabbits. The could occur. Death or serious illness would dog appears to be somewhat more suscep­ not be anticipated. 3) A daily dose of 500 m g./kg., if coutible to these materials than the other spe­ tinned for several day’s, could result iu cies studied, and chicks appear to be more tolerant. I t is also apparent th at MCP serious effects. A single dose of 500 m g ./ (2-incthyl-4-chlorophenoxy’acetic acid), sil- kg. would not be likely to cause serious vex (2-|2,4,5-trichlorophenoxy] propionic effects. 4) A single dose of 1,000 m g./kg. m ay or acid), and their derivatives are less toxic may not cause illness. Repeated doses of acutely thau the corresponding derivatives of 2,4-D and 2,4,5-T. The l.d.M values for this size are very likely to cause death. Evaluation o f Direct H atard to Live* 2,4-D and 2,4,5-T and their common deriva­ tives are iu the range of 300 to 1,000 m g./ stock.—In actual practice, esteron brush kg. fo r the rat, mouse, guinea pig, and rab ­ killer (a formulation of esters of 2,4-D anil bit, whereas fo r II CP and silvex, the corre­ 2,4,5-T) aud similar form ulations arc not sponding values range from 600 to 1,400 ordinarily used on areas containing appre­ ciable forage a t a rate greater th an 2 quarts m g.A g. The data presented in table 4 show th a t per acre or 47 mg./sq. f t Assuming that off of the herbicide ap­ the acute oral toxicity of the commercial plied a t this dosage rate is deposited ou formulations, in general, tends to be pro­ portional to th at which m ight be expected edible forage, an animal weighing 350 kg. from their content of active ingredients. (770 lb.) would have to consume all the Thus, it appears that the “ inert ingre­ forage on 744 sq. f t r f treated area to ac­ dients” present in these formulations arc quire a dose of 100 m g./kg. of the herbi­ less toxic than the active agents. F u rth e r­ cidal formulation. Such a dose could be more, the “ in erts” do not appear to exert tolerated daily for a number of days with­ a potentiating effect upon the toxicity of out any serious effects. Based on the same the active materials. F o r the commercial assumption, it would be necessary fo r the formulations studied, the Ld.M values for animal to graze completely an area ten rata, mice, gninea pigs, rabbits, and cattle times as large, or 7,440 sq. f t (0.17 acres) range from 500 to 2,000 m g ./k g .; for chicks in order to acquire an acutely toxic dose (approximately 1,000 m g./kg.). the range is from 2,000 to 4,000 m g./kg. Since these assumptions are based upon The results of repeated oral adm inistra­ tions indicate th at 2,4-D and 2,4,5-T can be complete availability* to livestock of all the tolerated without adverse effects in doses herbicide applied, an obviously impassible only slightly smaller than those which cause condition, it is apparent th at there is little, toxic effects when given only ouce. This if any, direct hazard to livestock o r wild­ fact demonstrates th at these m aterials have life foragiug areas treated with heabicides a low degree of chronicity. of the type described herein. This has been The studies with cattle have demon- confirmed by the extensive use of these mati*nial organs revealed no abnormalities. Histological examination of tissues from tlio lung, heart, liver, spleen, pancreas, thymus, bladder, lymph nodes, rumen. ret­ iculum, omasum, and abomasum revealed no. abnormalities. Very* slight interstitial edema and congestion was- observed in the m edulla and cortical m edullary regions of the kiduey, but since this has frequently been observed in untreated animals, its significance is questionable. I U O 0002824 O U O 5229 c G2S V. K. Howe and T. A. II ymas Icrials over a number of years w ithout any proved eases of adverse effects. - Eraluntion o f Indirect Hazard to Live­ stock.—There has been much discussion of the possibility th at spraying with these herbicides might cause some plants to be* come toxic, toxic plants to become more toxic, and ordinarily noupalatable plants to become palatable. -Considerable investi­ gation of these possibilities has resulted in the general conclusion th at from a practi­ cal standpoint these suppositions were more fanciful than factual. In 1950, W illard u of Ohio State Univer­ sity and A gricultural Experim ent Station . pointed out that the spraying of certain types of plants with 2,4-D increased their nitrate content to hazardous levels. The leaves of sugar beets accidentally ' sprayed with 2,4-D were reported to have caused the poisoning of some livestock. However, siuce 2,4-D is never knowingly used on sugar beets, this docs not appear to present a serious practical hazard. Other ■weeds such as lambsquarter, pigweed, and smart weed have also been reported as con­ taining increased quantities of n itra te after treatm ent with 2,4-D. Since livestock do not ordinarily cat these species, it would seem that the hazard presented by the ac.. cumulation of nitrate in these plants is ; slight. The lack of reported cases of nitrate ' / poisoning would seem to substantiate this supposition. Wild cherry is recognized as a hazardous plant to have in grazing areas. Couch* hits suggested th at the w ilting of wild cherry leaves results in the hydrolysis of cyanogenic glucosidcs to form free hydro: cyanic acid (IIC N ) and, therefore, th a t cut wild cherry is much more hazardous to live­ stock than the growing plants. By analogy, it has been postulated th a t wild cherry leaves wilted as a result of treatm ent with 2,4-D and 2,4,5-T herbicides likewise might be more toxic than growing leaves. Bar­ ren s and Lynn * and Grigsby and Boll,1* working, independently, d e m o n s tr a te d d e a rly th at the IIC N eontent of wild chcrxy leaves sprayed with estcron brush killer was actually lower than the content .msprayed leaves. Dr. W illard also has called to attention the possibility th at treatm ent w ith 2,4-D type herbicides m ight make certain natu­ rally poisonous plants more palatable or even attractive to livestock. This is a possi- Am. J. V kt. i : k». UCTIWM 1354 bility but, so far, it has not been estab­ lished ns a fact. In evaluating the hazard associated with the forementioned possibilities, Dr. Wil­ lard in 1951 ** stated as follows: “ In the first place, literally dozens of carefully controlled feediug experiments, both with 2,4-D and 2,4,5-T directly and with vege­ tation sprayed with them, have uniformly and without exception failed to show any poisonous effects on auy kind of livestock. “ In the second place, literally millions of acres of pastures with livestock on them have been sprayed in the last six or seven years without injury to livestock. I am sure there'is no experimenter here who has not used 2,4-D in occupied pastures. In our state, one company over a period of four years has sprayed 60,000 miles of roadside in twenty-six counties. D uring that time, he has had only five reports of suspected poisonings, none of them substantiated, 2 of swine, 2 of cattle, 1 of sheep. A company in Pennsylvania, which has sprayed 10,000 acres of right-of-ways p er year, reports ju st 4 cases, only 1 of which was not com­ pletely cleared up as due to other causes, in five years of work.” La 1953, Dr. S. N. P e r tig 1* of Cornell University summarized the situation re­ garding poisoning as follows, “ There are no known cases of actual herbieidal poison­ ing from field application of presently used h e rb ic id e s marketed as nonpoisonous.* There is no sound toxicological evidence available of the presently publicized ni­ trate-phenol and nitrite poisoning as a common cause of death. True, nitrate and nitrite poisoning has occurred in cattle, sheep, and swine from eating plants grown on high nitrate soils or from other feed sources. None of these, however, have been definitely traced or shown to be caused by herbieidal treatm en t All the alleged cases of herbieidal poisoniug of livestock and wildlife th at have been definitely diag­ nosed have bceu caused by one of the fol­ lowing: (1) lead, (2) arsenic, (3) hard­ ware disease, (4) poisouous plants, (5) old age, (6) parasites, (7) drowning, (8) poor marksmanship, (9) contaminated food, and (10) injection or oral dosage of some medi­ cine or drug. In all cases th at have been carefully surveyed, even though the herbi* D r . Turtle’« cUaufteatfea S t a r t h U M aurkrta4 M M -Iw iM m u ladaSaa rash aularisla aa Ibaaa Aiaaaaard la tfcia paprr. 5230 0002825 A uct¿ uïïi»*“' I nkjiimatiox on II ehbicides and an E valuation or T h km Lad eide has been associated with the trouble, it has in no case been directly or indirectly related to the deatlis reported.” ■ U» lad acress to spray tanks or other containers of the mutcrials. R ifin n c u . S ummary and Conclusions 1 Bucher, K uqr R.: KITerta ed M'OirUanpImiAcid on Rxprrimeatal AcimaU. P w . Sm. j 1 j The acute oral l.d.j, values for 2,4-D, exyaertic KxptL SM . u d Med, H , (1940): 304-303. 'HID. Kdiria C , end Cxrltxie. Harold: Toxicity ad ■\ 2,4,5-T, and their various derivatives com* DtcbleroplMaexyacetic Arid for Kxprrimraul Aaimonly used in hcrbicidal preparations fall 3.4mala. J. Iaduxt. Hy«. aad Toxicol, .'* , V* * r^, •• *■ ■ ** - -. -fc> - r i i > . F r ( - -J -T p-x ••-V, ■>,* -r -x,r v.v* v-iiAv Vr.TJv^# • ? Vi N^I^7* *>• V r > V ^ c f .* ":& T l % - KILLING SMALL UNDESIRABLE HARDWOODS by u« of the Co r n ell t o o l TABLE I By R a y E . G o d d a r d , Assistant Silviculturist, Texas Forest Service, A lto, Texas V o lu m b o f C h em ica l So l u t io n s a n d Average Siz e o f T rees T reated by B asal Spray and Co rn ell T o o l M ethods Applying 2,4 J - T in D iesel o il to hardwood with Cornell tool. The device is heavy, much like a spud, and permits the injec­ tion of the chemical in cuts in the basal area. Method of Application Basal spray Cornell tool Cornell tool Cornell tool Results of a small scale study, established in July and August, 1952, on the W . Goodrich Jones State Forest near Conroe, Texas, indicate that 2,4,5-T applied with a Cornell tree poisoning tool may be an effective method of releasing young pines from competing undesirable hardwoods. Plots, one-tenth acre in size, were set up to compare the effectiveness of 2,4,5-T ester applied as a basal spray and with a Cornell tooL The propylene glycol butyl ether ester was used in each case. A 2Vi per cent solution of 2,4,5-T, by volume, in diesel oil (10 lb. acid equivalent per 100 gal­ lons of spray), was applied by the basal, spray method to all hardwoods on two plots in July and August, 1952. Solu­ tions of 10 and 20 per cent 2,4,5-T in Diesel oil (10 and 20 percent 2,4,5-T, 40 and 80 lb. acid equivalent per 100 gal­ lons of spray) were applied with the Cornell tool to trees in two plots in July. In August, one additional plot was treated with a 10 per cent solution in Diesel oil and one plot was treated with a 10 per cent emulsion in water, both applied with the Cornell tooL As both application methods are primarily useful in the treatment of small trees, the plots were located in a stand dominated by sapling size hardwoods. However, the few larger hardwoods found on the plots were treated by the various methods being tested. Species predominating were redgum, blackgum and southern red oak. Smaller numbers of post, white knd water oaks, yaupon, holly, sassa­ fras, tree huckleberry,'red mulberry and red haw were .also found on the plots. W ith basal spray applications, the bark of treated trees was wet to the point of run-off from the root collar to Coacen' tration (b y volume) PerCent 2Vl 10 10 20 D iluent Diesel oil water Diesel oil Diesel oil Average Average volume used diameter of per tree* treated trees m l inches 200 8 3.7 2.6 3.2 3-3 11 11 * Aretwge determined from the total amount need per p lo t approximately one foot above the ground. Cornell tool cuts were spaced approximately one inch aparr around the tree base. Table I indicates average diameter of treated trees and the average volume of each solution used per tree. In September, 1953, a little over one year after treat­ ments were applied, 100 per cent of the hardwoods 10 inches in diameter and smaller treated with 20 per cent 2,4,5-T by the Cornell tool method were dead. Of these, only 1.6 per cent have resprouted. The 10 per cent 2,4,5-T in Diesel oil treatment was also effective. A total of 88 per cent of hardwoods under 10 inches in diameter were dead.. Hardwood mortality was less and sprouting more pro­ lific in the plot where die 10 per cent 2,4,5-T water emul­ sion was applied with the Cornell tooL Trees treated with 2,4^-T by die basal spray method usually die more slowly than when fresh wounds are treated. On die basal spray plots, 53 per cent of the hardwoods un­ der ten inches in diameter were dead one year after treat­ ment. By die start of the second spring after treatment, 63 per cent were dead. Many of the trees still surviving at that time were in a weakened condition and had been drastically reduced in living crown. ; .. The results of these treatments are summarized by species groups in Table U. As only two larger trees have died as a result of any of these treatments, only those less than ten inches in diameter have been included in these data. T a b l e ~IL'~-Percentage o f D ead H ardwoods 10 In c h e s d. b. h . and u n d er T w o Y ears a fter T r ea tm en t w it h _____________________. Basal Spray Co rn ell T ool Basal Spray T r eatm ents Spe c ies Redgum Bladegum Oaks* Yaupon Sc Holly Miscellaneous* All species 216% . 2.4.3-T in Dieiel OU Co r n e l l T o o l T reatm ents 10% 2.4.3-T in IVeter 10% 2.4.3-T m Jieiel Oil Number o f trees ¡reeled Per cont of trots dtod For cont of trots sprouting - Numbor of trots trostod Percent of ¡reel deed Forconi of trots sprouting Numbor of trots trootod For cout of trots iood Percent of trees tprouting 70 63 41 57 100 90 63 . 49 15 57 5 3 129 49 33 83 60 100 61 4 20 49 20 40 7 4 120 88 90 .88 100 75 88 0 17 14 6 11 117 0 0 12 0 0 2 23 0 0 17 0 3 0 0 1 20% 2.4.3-T in Dieiel Oil Number o f tree! . ¡reeled For cont of trots dood 100 47 100 10 ' 5..... "1 0 0 100 1 100 1 64 100 Percent of ¡reel sprouting 0 10 0 0 0 2 1 Includes mostly southern red oak with a few post, white and water oaks. *10010363 sassafras, wax myrtle, tree huckleberry, sumac, mulberry and. red haw. 5 5234 D o w n to E arth , F a ll 19. 3 5235 3 a _ a. APPENDIX V I I Determination of 2,4-Dichlorophenoxyacetic Acid (2,4-D) in Grain and Seed T ROLAND P. MARQUARDT and E. N. LUCE The Dow Chemical Co., Midland, Mich. The 2,4-D acid is extracted from a dry sample o f m eal with an acidified ether-chloroform solvent. By further treatment, including a chromatographic separation, interfering m aterials are removed. The color produced by the reaction of 2,4-D acid with chromo­ tropic acid in concentrated sulfuric acid is measured on a suitable spectrophotometer. As little as 5 y of 2,4-D acid .in q 200-gram sample can be detected. j r~ : i-Z /tk * 4 * i **■*■*• » __Jrw\ lie use o f 2,4-dichlorophenoxy a c e t ic a c id (2,4-D acid) as a weed killer on a variety of crops has made it desirable to develop an analytical method capable of detecting minute quantities of 2,4-D acid which may be picked up by the plant. The possibility of translocation of the 2,4-D acid must also be determined. The method of Marquardt and Luce (-/) was not adaptable directly to this problem, and the procedure of Gordon and Bcroza (o) did not ofier a means of separation from the substrata or indicate the possibility of detecting only trace nritirs. A method of extraction and separation of the 2,4-D acid was developed and the colorimetric method of Freed (2) further refined to give the accuracy demanded for this problem. R a a g en tt Chloroform, technical grade. Ether, U.S.P. grade. Acetic acid, glacial. Sodium hydroxide, approximately 52V. Hydrochloric acid, concentrated. So­ dium hydroxide, approximately 0.5V. Phosphotungstic acid. Dissolve 40 grams of phosphotungstic acid (approxi­ mately P jO,.24WO».25HjO) in water and dilute the solution to 100 ml. Hytlo Super-Cel, a Cclite product, diatomaceous silica, made by JohnsManville. Absolute methanol in U.S.P. chloro­ form, 3.0 and 7.0% by volume (see sec­ tion on chromatographic separation of 2,4-D acid). Buffered extraction solution, pH 6.85. Dissolve 10.0 grains of dibasic sodium phosphate, NajIIPO«.7HiO, and 10.0 grams of monobasic sodium phosphate, NaHjPOi.HjO, in water and dilute to exaedy 1.0 liter. Chloroform, analytical reagent grade. Ihromotropic acid. Dissolve 0.10 gram of the sodium salt of chromotropic acid (l,H-dihydroxy naphthalene-3,6-disulfonic acid) in 100 ml. of concentrated sulfuric acid (95.5%). Sulfuric acid-stannous chloride solu­ tion, dilute. Mix 200 ml. of concen- trated sulfuric acid with 800 mi. of water and cool the dilute acid to room tem­ perature; then dissolve 50 grains of stannous chloride, SnQj.2HjO, in the acid. 2,4-Dichlorophenoxyacctic acid, stand­ ard solution. Dissolve 0.500 gram of 2,4-D acid in chloroform and dilute to 500 ml. Dilute a 10.0-ml. aliquot to 100 ml. with chloroform. Dilute a 10.0-ml. aliquot of this second solution to 100 ml. with chloroform. The final solution contains 0.01 mg. of 2,4-D acid per ml. Apparatus The tube for the chromatographic column is a glass tube, 17 mm. in inside diameter and 50 cm. long, one end of which is constricted and attached to a smaller glass tube, 5 mm. in inside diam­ eter and 10 cm. long. A suitable apparatus for regulating air pressure and an interval timer are also required. A Coleman spectrophotometer, Model 11, equipped with a PC-4 filter and 5cm. cuvettes, was used in this investiga­ tion. Any photometer measuring light transmittance at 565 m u should be suit­ able. Proemdurn Grind the grain or seed to a fine meal to pass through a No. 10 sieve. In this work a No. 1 Wiley mill was used. Place a 200-gram sample of the meal in a 2-liter glass-stoppered bottle, and add 500 ml. of technical grade chloro­ form, 500 ml. of ether, and 10 ml. of acetic acid. Stopper the bottle, fasten­ ing the stopper with Okonite tape. Place the bottle and contents in a shaking machine for 1 hour. Use a 2-liter filtering flask, a Büchner funnel 16 cm. in diameter, and What­ man’s No. 1 or similar grade filter paper. Rinse the bottle by adding 50 ml. of technical grade chloroform and 50 mi. of ether and wash the insoluble material with the rinsing solution. Discard the insoluble materiaL Using a 2-liter separatory funnel, extract the 2,4-D acid from the filtrate once with a solution of 200 ml. of water and 50 ml. of 5V sodium hydroxide and twice with a solution of 200 ml. of water and 10 ml. of 5V sodium hydroxide. Shake well each time to ensure complete extractions. Discard the organic layer and wash the combined aqueous extracts three times with 100-ml. portions of technical grade chloroform. Acidify the aqueous solution with 50 ml. of concentrated hydrochloric acid. Extract the 2,4-D acid from the solution with 125 ml. of ether. Repeat the ex­ traction with 100 ml. of ether and com­ bine the extracts. Discard the aqueous solution. Filter the ether solution, using What­ man’s No. l" or equivalent grade filter paper, into a 250-mi. separatory funnel. Extract the 2,4-D acid from the filtered solution once with 50 ml. of 0.5V sodium hydroxide and twice with 25-mi. portions of 0.5V sodium hydroxide. Combine the caustic extracts. Pour the caustic solution into the 250ml. separatory funnel and wash it three • times with 10-mi. portions of chloroform, discarding the washings. Acidify the solution with 10 ml. of concentrated hydrochloric add and add 5 ml. of phosphotungstic add solution. Extract the 2,4-D acid with three 10-ml. portions of chloroform and combine the extracts. Prepare the column for the chromato­ graphic separation by placing a plug of glass wool in the bottom of the tube at the constriction. Add Hyflo Super-Cel and settle it in the tube by bouncing the tube gently on a large rubber stopper. Continue adding more Super-Cel and bouncing the tube gently until the SuperCel is packed in the tube for a distance of 20 cm. Filter the chloroform solution into the tube, using No. 1 Whatman’s or e q u iv a ­ lent grade filter paper. Apply air pres­ sure at the top of the tube to push the solution into the Super-Cel. Using proper air pressure, percolate 150 ml. of the 3.0% methanol in chloro­ form solution through the column at the rate of 150 ml. per hour, followed ^ ’^ 5 ^ g Reprinted from AGRICULTURAL AND FOOD CHEMISTRY, Vol. 3, No. 1, Page 61, January 1955 n n m 3 * g Q Tn n n KERBCIDE DETERMINATION It o acre (as low volatile ester) should be applied after plants have reached full leaf in the spring. The cost will be about $8.00 per acre but these oak areas have more favorable rain­ fall and will produce more forage than many mesquite areas. It should be emphasized that for both mesquite and oak, control rather than eradication is die result. Additional sprayings will be necesary to control regrowth. Can all kinds or upland and bottomland hardwoods be aerially con­ trolled economically? There is a need for more research on this problem. Aerial application is die cheapest method known with the limitation that acreages must be large and no susceptible crops are dose by. Possibly hardwoods may be controlled by aerial applica­ tion of 2,4,5-T, silvex, or MCP in timber areas. Earlier re­ search indicated chemicals would kill pines but later work makes researchers wonder. These possibilities should be fully explored. Chemicals may be applied to the foliage of individual trees, to the cut-off stump, trunk base of standing trees or in cups or frills made in the bark 2,4,5-T ester and ammon­ ium sulfamate are the most widely used and accepted chem­ icals. In Texas and Oklahoma, 2,4,5-T has generally given cheaper and more effective results with less labor. Stump treatment using 2,4,5-T ester diluted in diesel oil or kerosene gives good control of sprouting for most spe­ cies (Figure 2). Trunk base treatment using 2,4,5-T in oil gives good results on trees under six inches in diameter but some species require twice the concentration of chem­ ical as others (Figure 3). Ammonium sulfamate crystals in cups spaced not over six inches apart and close to the ground or applied in water solution to frills usually con­ trols sprouting on oaks. A solution of 2,4,5-T in oil ap­ plied in single overlapping axe cuts shows much promise of cheap, effective control of all kinds of trees. The value of any program is dependent upon its accep­ tance by the people. Extension woody .plant control in Texas has been presented through county agent training meetings, method demonstrations in counties, newspaper, magazine and radio stories and TV demonstrations. In 1952, working with the county agents, the writer conducted brush control meetings or demonstrations in 46 different counties. In 25 counties, 33 field demonstrations were set up using 2,4,5-T and am m o n iu m sulfamate on stumps, trunks and in frills. These demonstrations were perman­ ently marked with the kinds of chemical used and method of treatment. Last year, 40 brush control meetings and 30 field demonstrations in 24 different counties were held. For the past two years, this gives a total of 86 brush con­ trol meetings, 63 field demonstrations in 49 different coun­ ties and 150 hardwoods demonstrations conducted by county agricultural agents and this specialise As a result of an evaluation of these field demonstrations together with experiment station results, a circular, "More Grass from Controlling Hardwoods with Chemicals" C-330, has been prepared for the guidance of Texas farmers and ranchmen and is available from the Texas Agricultural Ex­ tension Service. A Ch a llen g e Whereas brush control is the biggest problem facing the farmer, ranchman and timber producer in many southern areas, it also presents a very great opportunity. In busi­ ness 3 or 4 percent interest is considered a fair investment while the application of present woody plant control infor­ mation can result in 20 to 50 per cent increase in desirable production and 100 per cent gain is not uncommon when coupled with proper range, pasture and timber management. mended exclusively. Operators should study their particu­ lar conditions in die light of available information to de­ termine the most profitable methods to follow. ost chemical research work on woody plant control u.. range or pasture land should be guided by these funda­ mental objectives: (1 ) development of effective treatment, (2 ) cost and possible net return and (3) range manage­ ment practices necessary for continued benefits. Some fed­ eral and state research men who have contributed materi­ ally to chemical brush control in the South are: C E. Fisher at Spur, Texas on mesquite; Dave Savage and Pat Mcllvain at Woodward, Oklahoma on sagebrush and Shinnery oak; Fred Peevy and John Cassady on Louisiana oaks; L E. Chaiken on hardwoods in North Carolina; Harry Elwell on oak brush at Guthrie, Oklahoma; and R. A. Darrow and Wayne McCulIy on post and blackjack oaks, prickly pear, whitebrush and huisache in Texas. This work has guided further research and demonstrations in the South. Chemicals used in brush control work may be selective such as 2,4-D, 2,4,5-T and silvex or nonselective such as ammonium sulfamate, arsenic and kerosene. Fundamentals of the ultimate in a woody plant control chemical are: (1) nonpoisonous to man or livestock; (2 ) effective under a wide range of conditions; (3 ) cheap; (4) easy to apply and to handle; (5 ) not damaging to grass or cultivated crops. Present chemicals meet some of these requirements but not all of them. Chemicals may be applied by broadcast methods or by individual tree or plant treatment. Tractor or jeep drawn power spray equipment can be used on low growing brush or following mechanical control work. Aerial application provides economical and rapid coverage of large brush areas. Such broadcast methods have certain limitations. At the;ent time, most chemicals applied when the brush is in . foliage, control only certain kinds of brush and must be used very carefully in susceptible crop areas. Sprays may be applied to individual plants with hand operated or cattle sprayers. Such equipment is suitable for foliage treatment of scattered individual seedlings or small brush and for frill, basal and stump treatment. Extensive brush control research conducted at the U. S. Southern Great Plains Field Station at Woodward, Oklahoma shows that three-fourths of the sand sage­ brush can be killed with one proper aerial application of 2,4-D. Forage and beef production is increased from 50 to 75 percent if grazing is light or summer deferment is prac­ ticed following treatment. The cost of sagebrush spray­ ing on experimental areas has been repaid completely the first year following treatment. More than one million acres of mesquite have been aeri­ ally sprayed in Texas during the past 3 years with Yi -Va lb. of 2,4,5-T (as low-volatile ester) per acre in diesel oil at a cost of $3.00 to $3.50 per acre. Top kills will average around 90 percent and root kills 40 percent. Mesquite is the biggest one species problem in Texas with a dense stand on 35 million acres and scattered stands on another 15 million acres. It is a prolific sprouter from the bud zone. Stage of growth, moisture condition, size of trees and m an, agement following treatment largely determine the bene­ fits derived from spraying. Grass growth and beef produc­ tion have been materially increased on all areas sprayed and the writer has yet to visit an area on which mesquite sprayi° " has not paid for itself. erial control of post oak and blackjack oak has been tried on a limited scale. It appears practical and economi­ cal (Figure 1). Two pounds of 2,4,5-T add equivalent per ii 5237 D o w n to E arth , Summer 1954 3 o 3o 5238 C -T - • * m Summary o f Toxicological Information on 2,4-D and 2,4,5-T Type Herbicides and an Evaluation of the Hazards to Livestock Associated with Their Use V. JC ROWE, M.S.. and T. A. HYMAS, D.V.M. Midland, Michigan cnee in potency I«tween crude and purified projw- DOW 5087F3 T h e rs E o r herbicide* for tiie control of undesirable vegetation lias been practiced rations, or lictween tiui sodium or ammonium salts. for many years. Some of the older mate* Deaths from large dose* were believed due to ven­ riaU commonly used are chlorates, arseni- tricular fibrillation; if death wan delayed, myoto­ nia. stiffness of extremities, ataxia, paralysis, aad cals, compounds of borou. certain oils, and cornu were olamrvedr even table salt. While many of these mate* In Kulmeutr studies, the wine investigators ol>rials are still being used for this purpose, served intoxication in dogs after six daily intra­ research over the past decade has revealed venous injections of 22 GnuAg.; rata were fed a a family of new herbicidal materials. They diet containing 1.000 parts per million (p.p.ra.) of are the chlorinated phenoxvacetic acids and 2.4- D for a month without harmful effects; guinea closely associated compounds -of which pigs tolerate*! ten doses of 100 m gA g- over a 2.4- D (2,4-dichlorophenoxyucetic acid) and twelrc-dny jieriod; aad the inhalation of the so­ w it as a dost failed to eanso systemie effects 2.4,5-T (2,4,5-trichIorophenoxyacetic acid) dium in guinea pigs. The main evidences of ehronie in­ are prototypes. toxication oliserved in rats were visceral conges­ Early in the development of these new tion an*l edematous kidneys with degenerative herbicides, studies were undertaken by The ehnngcs ia the tubules; only dogs exhibited hepatic Dow Chemical Company to determine their damage with central degeneration aad congestion. Ia 104», Bjorn and N’orthcn' reported the re­ toxicity to warm-blooded animals. I t is the purpose of this paper to make the acquired sults of studies conducted on ehieks with an alka-. toxicological information a v a ila b le to uolamine w it of 2,4-D. Tho acute oral lethal rang« was found to be 380 to 762 mg.Ag. When given others. V repeatedly (12 doses ia 28 days), 28 mgAg-/doso waa without effect, while 280 mg-Ag./dose caused depression of growth. These authors express no Uucherl ia 1046 wan among tlx* Ant to report concern about the likelihood of tosic effects ia the reanlU of experiment* with »mail anirnul* using ehirkeus under ordinary conditions of nao and 2.4- D. Temporary myotonia lasting from eight to point ont that at a spraying rate of 1 lb. of 2,4-D twenty-four hours or more following a single in­ per acre (a normal rate of application) a ehiekea jection of ISO to 250 mg./fcg. wa* observed' in weighing 1 kg. would have to consume all the miee. rats, rabbits, and dags. Repented injections 2.4- D applied on 72 sq. ft. within a day or two to o f «mailer amounts, 50 to 100 mg.Ag./day to mire ubtuin a lethal dose. for ninoty days, failed to elicit either n character­ Ia 1P55, Drill and llirutska' reported the results istic chronic syndrome or n striking histological of acute and ehronie oral toxicity studieo on 2,4*1) picture. Miee undergoing this treatment became and 2,4,5-T with dogs. These men found that the pregnant and bore apparently normal litters. Re- acute oral l.du> values wera about 100 mg-Ag. far IK-atcd injections of 2,4-D did not alter the rate of each of the material*. At this dosage level, 2,4-D growth of twu transplantable mouse sarcomas. produced definite myotonia accompanied by anor­ In 1047, Hill and Carlisle* published the results exia and weight low while 2,4,5-T produced only of toxicological studies involving various prepara­ signs of mild spasticity. Both 2,4-D and 2,4,5-T tions of 2,4-D. In sente oral studies, they found were fed five times a week for ninety days at the L i s for miee to be 375 mgykg.; for rata, G68 dosage levels of 2, 5, sod 10 mgVkg. without ad­ utg-Ag.; for rabbits, MOO mgykg.; and for guinea verse effects. At a dosage level of 20 m itA a , pigs, 1,000 mgVkg. The hugest dose administered Imtli materials caused serious effects; 3 of 4 dogs to monkeys without serious after-effeeta was 214 on the 2.4-T> died while oil of tho 4 dogs on 2,4,5-T mg-Ag.; 428 mg-Ag. roused nausea, vomiting, died. The animals receiving tho 2,4-0 exhibited lethargy, musel# incoordination, aad head drop. stiffuem of the hind legs, difficulty in swallowing, These workers obserrsd that all species m eted bleeding of the gums, ueerorie ehangeo in the similarly and that there was no significant differ- lmerul mueow, and mild liver and kidney changes. A significant decrease la the number of blood Vrotm the Biochemical Besrarrh Départant (Bowel aad the Acricultural Chrsikal Rasesreh Laboratory lyntphoeytes was observed terminally ia 3 of the 4 animals. The toxic symptoms noted in the animals (Kjioss), The Dow Chemical Co^ MMIaad. )(irh. Unaumnuc Review [822] 5239 : 0002319 Calculation Calculate the parts per million of 2,4-D acid present in the sample as follows: Mg. of 2,4-D acid X 1000 grains of sample p.p.m. of 2,4-D add Preparation of Graph Add 0.0, 1.0, 5.0, and 10.0 ml. of the standard solution containing 0.01 mg. of 2.4- D acid per ml. to 200-gram samples of untreated meal, and determine 2,4-D acid according to the procedure. Using the data obtained, construct a suitable graph. Data obtained by the authors on wheat meal are shown in 1 able I. Reference data on known amounts of 2.4- D acid were obtained by pipetting 0, 1.0, 5.0, and 10.0 ml. of the standard solution containing 0.01 mg. of 2,4-D ad d per ml. into four 50-ml. beakers and gently evaporating the chloroform by­ use of a steam bath. Then 5.0 ml. of chromotropic add reagent were added and the color was developed according to the procedure. Results are given in Table II. The data from Tables I and II are shown graphically in Figure 1. Com­ parison of the two sets of data shows that the recovery of 2,4-D acid, although not complete, is adequate to detect and esti­ mate trace quantities of the acid in wheat meal. The over-all loss is corrected in the preparation of the graph. Graphs made for meals of barley’, flax, dried peas, and oats were similar to the one prepared for wheat meal. For oats, it was fouQd expedient to use 100-gram samples. The analytical data found on the samples of grain and seed listed above correlate very well with the information available in each case. Chromatographic Separation o f 2 ,4 -0 A cid two The developing solution and the elut­ ing solution used with the Hyflo SuperCel for the chromatographic separation of the 2,4-D acid should be carefully standardized in order to make a proper separation. The adsorptive strength of Hyflo Super-Cel was found to vary somewhat ^ 1* Table 1. Transm ittancy V alues for 2,4-D Acid in W heat Meal % rroflsmrtfoftcy ¿ ,4 -0 Acid, Mg. 89.0 Nil (blank) 79.8 0.01 58.8 0.05 42.0 0.10 Tabla II. Transmittancy Values for 2,4-D Acid* 3 ,4 -0 Add, Mg. % TrorumiHoocy Nil (blank) 0.01 0.05 0.10 97.1 86.2 53.7 27.8 from batch to batch. Therefore, a uni­ form batch of Super-Cel, in adequate supply for standardization of the chro­ matographic separation and for use in many subsequent separations, should be set aside. The presence of water was found to affect the elution strength of the solu­ tions used in the chromatographic sepa­ ration. Since the water content in tech­ nical methanol may vary from drum to drum, several gallons of methanol from a selected drum should be stored and used for standardizing the solutions and for the duplication of these solutions there­ after. In this investigation, methanol containing 0.03% water was used. It is probably not necessary to store chloroform, as variation in quality from various lots appeared to have no signifi­ cant effect on the elution strengths of the solutions. Prepare the column for the chromato­ graphic separation by packing the tube with Super-Ce] for a distance of 20 cm. Pour about 30 ml. of chloroform con­ taining 0.10 mg. of 2,4-D arid into the tube. Apply air pressure at the top of the tul>e to force the solution into the Super-Cel. Percolate 150 ml. of 3.0% methanol in chloroform solution through die column at the rate of 150 ml. per hour, followed by 25 ml. of 7.0% chloroform in meth­ anol. Discard the effluent. Then pass 50 m>. of 7.0% methanol in chloroform through the column at the same rate and save the liquid in a 100-ml. beaker. Repeat this tlirce times with the similar portions of the eluting sulution. Test for 2,4-D acid in each beaker as follows: Evaporate the liquid by use of a steatn bath, add 10 ml. of chromotropic acid reagent, and swiri to make a uni­ form solution with any residue. Piece the beaker and contents in an oven set at 150° ± 2s C. for 10 minutes. Then examine the solutions for the wine-purple color which indicates the presence of 2,4D acid. For a good chromatographic separa­ tion of the 2,4-D acid, results should be: _ „ _ , 509189 Figvre 1. Recovery of 2,4-D odd from wheat moot stannous chloride, solution. Mix well and (¡her through a Whatman’s No. 42 or equivalent grade of filter paper. After 30 to 45 minutes, determine the per cent transmhiancy of the filtrate at 565 ma, using 5-cin. cuvettes with water as the reference liquid set at 100%. Read the milligrams of 2,4-D acid contained by the sample front a graph made front the data obtained by analysis of untreated meal to which known quan­ tities of 2,4-D acid had been added. DOW ml. oi 7.11% methanol in chloroform. Discard llir clllucnt. Then percolate 15» ml. of 7.0% meth­ anol in dilornforiu through ilic column at the same rate, collecting the cfllucnt in a 250-inJ. beaker. Pour the diluent into a 250-ml. separa­ tory funnel. Extract the 2,4-D acid from the solution with three 25-mi. por­ tions of buffered extraction solution and combine the extracts. Return the extract solution to the separatory funnel and wash it three times with 10-ml. portions of analytical reagent grade dtloroform. Discard the wash­ ings. Acidify the solution with 2 ml. of con­ centrated hydrochloric acid and extract the 2,4-D acid with three 10-inl. portions of analytical reagent grade dtloroform. Combine the extracts in a 30-ml. beaker. Decant the chloroform solution from the globules of water adhering to the beaker into another clean, dry 30-ml. beaker. Gently evaporate the chloro­ form by use of a steam bath (test tube clamps may he used to hold the beakers d rrp in the steam bath to facilitate the evaporation). Evaporate just to dry­ ness and remove the beaker immediately. Cool. Add 5.0 ml. of chrnmwmpic acid re­ agent to the residue in the beaker and swirl to make a homogeneous solution. Place the lieakcr and contents in an oven set at 150s =fc 2* C. for exactly 10.0 minutes (use interval timer). Cool the solution to room temperature (winepurple colored if 2,4-D is present). Pour the solution with stirring into about 30 ml. of sulfuric acid-stannous chloride solution, rinsing the beaker with a few milliliters of the solution. Stan timing with the interval timer and cool the solution to approximately room tem­ perature, using a cold water bath. After abuut 5 minutrs, pour the solu­ tion into a 50-inl. volumetric flask and make tu volume with sulfuric acid- H H flO 0 ri 1 Discussion Phosphotungstic acid is used to sepa- ' rate proteins when the 2,4-D acid is ex­ tracted with chloroform from the acidi­ fied aqueous solution. The buffered extraction solution sepa­ the desired compound. The procedure as given has been used to determine 4chloro-o-toloxyacetic acid (MCI* acid). Although this procedure was developed principally for the determination of 2,4D acid in grain and seed, it can probably *be used for other agricultural products. Fresh vegetables such as peas should be dried by suitable means and the analysis made on a meal of the dried sample. It is possible to detect less than 0.05 p.p.m. of 2,4-D acid in samples by this analytical procedure. Interferences by other compounds were not experienced as, under the conditions of the procedure, the color test is specific for aryloxyacedc acids. LHorxdurw CHod (1) Bricker, C. E., and Johnson, H. R., Ind. Eng. C km ., Anal. E d., 17, 4003 (1945). (2) Freed, V. H., Scitnct, 107, 98-9 (1948). (3) Gordon, N., and Beroza, M., Anal. C km ., 24, 1968 (1952). (4) Marquardt, R. P., and Luce, E. N., Ibid., 23, 1484-6 (1951). R a tia td fo r rttruw Ja m m y 20, 1954. coptod S n tm b tr 8 , 1954. MOa The fourth 50-tnl. portion of 7.0% methanol in chloroform is not run through the tube during a regular analy­ sis. It is done here for testing the chromatographic separation to make sure that all of the 2,4-D acid is eluted with the first 150 ml. of 7.0% methanol in chloroform. If the results do not show a good sepa­ ration of the 2,4-D acid, the amount of methanol in the two solutions of chloro­ form should be adjusted. rates the 2,4-D acid from some of the small amount of acidic material remain­ ing with the 2,4-D acid after the chro­ matographic separation. After the final separation of the 2,4-D acid, a very small amount of acidic material from the grain sample may be present, which will give an amber color when healed with chromotmpic acid reagent. The stannous chloride in the dilute sulfuric acid bleaches the amber color, but under the conditions of the experiment it docs not bleach the winepurple color. Though the solution from an untreated grain sample obtained by analysis is not water-white, the per cent transmittancy is high and constant. The color reaction of 2,4-D acid with chromotropic acid is not quite specific. Formaldehyde will produce the same wine-purple color (7). According to Freed (2), phenoxyacetic acid and its derivatives will give the same color, perhaps because formaldehyde is a de­ composition product. The procedure can probably be used for phenoxyacetic acid and many of its derivatives besides 2,4-D acid, although the solutions of methanol in chloroform may have to be modified to make a proper chromatographic separation of 06I60S 1. At must, a faint w ine-purple color in the first beaker, indicating a trace of 2,4 -D acid. 2. A deep w ine-purple co lor in the icco n d beaker, indicating ino»t of the o rig inal 0.10 mg. o f 2 ,4 -0 acid. 3. A pale w ine-purple color in the third a k cr, indicating only a sm a ll portion of m e orig inal 0.10 mg. of 2,4 -D acid. 4. N o w ine-purple color noticeable in the fourth beaker, indicating the absence of 2 ,4 -D a cid . Ac- a 0002832 33 5242 w m * T H È STATUS Of ? £ $ T 1 C 1 D £ S ü M O e f t T H £ M tL L tA *m €Nom M r o THE. F e& £ ftA i poop,OlíJJG 4 c o i e r i e G-E Ly n n , The Dow Chemical Company A C -r at the present time and on which there seems to be con­ siderable confusion. 1. Some products are exempt from the provisions of the Law because they are not considered poisonous or dele­ terious. Example: Sulphur. 2. Products which have no tolerance established. This category would include materials which leave no residue and, therefore, require no tolerance. Example: 2,4-D on small grains. . 3. An exemption from a tolerance. Food and Drug may exempt materials from a tolerance because of their safe characteristics. Examples: Pyrethrum and Copper. 4. A product may have a tolerance established in so many -parts per million. Example: DDT, which has a seven parts per million tolerance. 5. A zero tolerance. A zero tolerance would apply when a material can be used but only under such conditions as result in no residue when the crop is marketed. Example: Mercury. A zero tolerance is often confused with "no tolerance.” 6. Certain provisions of the Act may be extended until July 22, 1956. "Those interested in finding out where a product stands in the six categories listed above should write to the manufacturer." Although representatives of industry and Government have tried to clarify the status of pesticide chemicals under die Miller Amendment (Public Law 518, 83rd Congress) to the Federal Food, Drug and Cosmetic Act, there are still some misunderstandings regarding recommendations that can be made for the coming season. Mr. L S. Hitchner, Executive Secretary of the National Agricultural Chemi­ cals Association, speaking on the subject at The Confer­ ence on Problems Involved in the Study, Evaluation and Application o f Pesticides with Special Reference to Safety of Use, Washington, D. G, October 19, 1955, said in pan: "Under present Federal legislation, industry is required to supply two basic types of data before sale in inter­ state commerce. 1. To the United States Department of Agriculture, information and evidence showing the effectiveness of the material and of particular interest at this time the pesticide residue remaining on a crop when used in accordance with label directions. This information, in due course, reaches, where a residue remains, the Food and Drug Administration. 2. To the Food and Drug Administration, the toxicity daca required by them in order to determine a tolerance, if one is required. In establishing a tolerance the amount of residue remaining is considered. "The residue data is obtained from work done by com­ panies in their own laboratories; from held operations; from the work of Land Grant Colleges, both on cooperative proj­ ects and independently; from private research institutions and from expert research organizations doing work in toxi­ cology and medicine. "The result of this program means that adequate infor­ mation on residues is deposited by the company to reason­ ably assure, that when used as directed, residues, if any, will be within the tolerance determined by the Food and Drug Administration. Under this program the grower is reasonably assured that when used as directed there is no • excess residue and the public likewise is assured there is no undue hazard to the public health. "As a practical matter, growers and others will have to ' depend primarily on the label to assure there is no excess residue. All instructions on the label, including directions for use, timing of applications and other conditions must be explicitly followed.” Mr. Hitchner also pointed our that there were, "six cate­ gories in which the residue tolerance problem can fall Status o f D o w A gricultural Ch em ica ls All current labels for Dow herbicides, insecticides, fumigants and fungicides have been cleared by the Pesticide Regulation Section of the United States Department of Agriculture. In those cases in the following text where extensions are indicated it is probable that tolerances or exemptions will be established before the law becomes fully effective on July 22, 1956, and that current label directions will continue to be in effect. H erbicides 1. Baron (2-(2,4,5-trichlorophenoxy)ethyl 2,2-dichloropropionate). Not presently used on food crops, therefore, no tolerance is necessary. 2. Brush Killer T, Brush Killer 50-50, Esteron Brush Killer, Esteron 245, and Kuron (2,4-D, 2,4,5-T and silvex [2,4,5-trichlorophenoxypropionic acid] brush killers). Not 8 5243 used on food crops, with the possible exception that some might be sprayed on pastures in areas immediately around brush. No residue problem exists (see 3) and no toler­ ances are necessary. 3. All 2,4-D and 2,4,5-T weed killers. Data have been presented to the ILS.D-A. that show no residue problem exists on small grain or in milk. In addition, the F.DA. has agreed that there is no residue problem in meat. No residue tolerance is necessary. 4. Dow MGP Amine Weed Killer, (4-chIoro-o-toIoxyacetic acid, or 2-methyl-4-chlorophenoxyacetic acid, amine salt). The character and use of die chemical are similar to 2,4-D. It is the opinion of qualified experts in the field that no residue problem exists and that no tolerance will be necessary. An Industry-National Agricultural Chemi­ cals Association Committee is now conducting analyti­ cal investigations. 5. Dalapon, Sodium Salt 83%, (sodium 2,2-dichloropropionate). Data have been presented to the U.S.D.A. that show no residue is present in sugar cane at harvest. With respect to pre-planting applications, dalapon is destroyed in warm, moist soil within several weeks after application (see D o w n to E arth , 11 (2 ):2 (1955) ). 6. Dow General Weed Killer, Premerge and Dow Selective Weed Killer, (formulations based on dinitro-orec-butylphenol). Dinitro-o-rec-butylphenol has a zero tol­ erance. Dow has presented data to the U.S.D.A. that show no residues are present in the harvested crops when the products are used in accordance with label directions. These have been cleared by the U.S.D.A. It should be noted that in pre-harvest treatment of seed crops with Dow General Weed Killer, the label warns that the forage is not j o be used as animal food, since such forage may contain DN residue. 7. Sodium TCA 90%, (sodium trichloroacetate). Data have been submitted to die U.S.D.A. that show no resi­ dues are present in sugar beets, cabbage and tomatoes when used in accordance with label recommendations. In view of the sugar cane data on dalapon, it is unlikely that residues will occur in this crop. Data are now being obtained. It is not anticipated that residue tolerance will be necessary for TCA. 14. Lindane, BHG Tolerances for a large number of, raw agricultural commodities have been set as follows: l i n d a n e ............................. 10 ppm benzene hexachloride . . 5 ppm An extension has been granted for lindane for several additional items until January 22, 1956 and on BHC in meat until March 1, 1956. 15. Lime Sulfur, Sulfur. The residues left by these pestiddes are exempted from a tolerance. 16. Ovotran Wettable (p-chlorophenyl p-chlorobenzenesulfonate). An extension has been granted until March 1, 1956 for citrus. A petition has been submined for tol­ erance in and on dtrus, apples, pears, peaches, plums and prunes. 17. Parathion. A tolerance of 1 ppm has been set for a wide range of fruits and vegetables appearing on the Dow labeL An extension has been granted to January 22, 1956 for field CTops, forage aops, hops and olives. 18. Systox. A tolerance of 0.75 ppm has been set for apples, broccoli, brussels sprouts, cabbage, cauliflower, muskmelons, oranges, pears, potatoes, strawberries and walnuts. 19- Dowfume 75, Dowfume EB-5, Dowfume EB-15, Grain Fumigant (80-20 Mixture). All grain fumigants con­ taining carbon tetrachloride, carbon disulfide, ethylene dichloride, and ethylene dibromide have been extended until March 1, 1956. Dow and other companies have been doing analytical work with the Food and Drug Administra­ tion to obtain the necessary data to establish tolerances or exemptions. 20. Methyl Bromide. An extension has been granted until January 22, 1956 for certain raw agricultural com­ modities. Dow has filed a petition asking for tolerances on apples, pears, quinces, onions, tomatoes, eggplants, beets, beans, cocoa beans, wheat, rice, rye, oats, barley, com, grain sorghum, sweet potatoes, alfalfa hay, peanuts, potatoes, pecans, turnips, rutabagas and cottonseed. 21. Dowfume W-85, Dowfume W-40, Garden Dowfume ( ethylene dibromide soil fumigants). An extension has been granted until January 22, 1956. Dow has filed a petition asking for tolerances on: Lima beans, com, strawberries, sugar beets, asparagus, lettuce, sweet potatoes, parsnips, potatoes, turnips, rutabagas, carrots, celery and cottonseed. (Tobacco was not requested since it is not a food). Addi­ tional data are being obtained for tolerance in other aops. Insecticides 8. Arsenare of Lead. A residue tolerance of 7 ppm of combined lead has been set on the raw agricultural com­ modities that appear on the Dow labeL 9. DDT. A residue tolerance of 7 ppm has been set on a wide variety of raw agricultural commodities appearing on the Dow labeL An extension has been granted for meat until March 1, 1956. 10. DN-Dry Mix No. 1 (dinitro-o-cydohexylphenol). A residue tolerance of 1 ppm on citrus has been set. 11. DN-Dry Mix No. 2 (dinitro-o-cresol). A zero tolerance has been set. No residues from stria dormant application. 12. DN-289 (dinitro-o-xec-butylphenol, triethanolamine salt). Zero tolerance. No residues from strict dormant application, or when applied after harvest. 13. DN-111 (dinitro-o-cyclohexylphenol, dicydohexylamine salt). A 1 ppm tolerance has been set on apples, apricots, beans, blackberries, black-eyed peas, celery, cherries, citrus, grapes, loganberries, neaarines, peaches, pears, plums, quinces, raspberries and strawberries. F ungicides 22. Lime Sulfur, Sulfur (See item 15). 23. Ferradow (ferbam). A tolerance of 7 ppm has been set for a wide variety of fruits and vegetables appearing on the Dow labeL O th er A gricultural Ch em ica ls Nematocides, defoliants, and plant growth regulators are not defined as economic poisons by the Federal Inseaicide, Fungicide, and Rodenticide A a. They are not registered by die U. S. D. A. and tolerances cannot be set under the Miller Amendment. Tolerances of 1 ppm naphthaleneacetic acid (App-L-Set) and 5 ppm for 2,4-D were s a on certain fruits as a result of the 1950 hearings. Data are being obtained on Color-Set (2,4,5-trichlorophenoxypropionic acid) to determine if residues - are present at harvest. Magron (Dow magnesium chlorate defoliant) has been investigated and no residues of magnesium chlor­ ate have been found on rice or beans defoliated with this produa. 9 5244 D o w n t o Earth , Winter 195 34 5245 i T ...A.- 1 Containing 6.15 Pounds per FOR THE Ac||ve Ingredient: 2,4,5-Trtchlorophenoxyacetlc Acid, Propylene Glycol ( C | l l , | ) Ip c ( l l , <0 j l Buryi Ether E s t e r s __________ _____________________ 2,4,5-Trichlorophenoayacellc Acid Equivalent 42.5?» Inert In g re d ie n ts____________________________________________________________________ «5.3 34.7% Esteron 2 4 J it recommended for use in controlling herboceous and woody plants grow­ ing in rangeland, pastures, fence rows, ditch banks, farmyards and right-of-ways. It is cffectise in controlling certain 2 ,4 -0 resistant plants such as ash, brombles, ground chefty, hawthorn, horse nettle, maple, mesquitc, oak, osage oronge, palmetto, poison ivy, prickly pear cactus, salmonbcrry, wild blackberry, wild rose and certain species of Ribcs, as well as many other woody ond herboceous species. D IRECTIO N S PREPARIN G THE S P R A Y : Add holf the required amount ol water or oil to the spray lank, then odd the E.tcrnn 245 with oqilalion, and finally the bolancc of the water or oil with continued oqilolipn. W A R N IN G : II Esteron 245 is to be used In preparing straight oil mi «lures, «la not let «voter get into the Esteron 245 itself nor into lire fin­ ished m ivlqic. N O T E: Esteron 2 15 ferms on emulsion— not a solution - in water, and the Esteron 245 lends to separate out on slandinq. Provide agitation to prevent such separation and ensure uniformity of spray mixture. For Trealinq Smeli A rc a i: One lahlcspoonlul cf Eslcron 245 in l ' j «voler is a|-.v''ximatrly eguivalent to onc quoti in 100 galloni. gotico« ol oil or FO LIA G E T R E A T M EN T : Spray woody growth up to 6 or 8 fuel toll alter foliage is well developed, using o d r e n c h i n g sproy containing 3 quarts of Eslcron 245 per 100 gallons ol «safer. Toller brush can be sprayed successfully, although in many cases basal bark or stump treatment is prcfcroble. Poison Ivy, most brambles and some other species may be controlled bv using 2 quarts per 100 gallons of water. Coverage should ha 'omplele, ond oil ports of the plants, Including foliage, shoot stems ond bark, should be wet with the spray, Bed results usually w ill be obtained from applications made scon o iler maximum lalioge development in the spring. W ith good growing conditions ond adequate soil moisture, applications may l,e mode up to 2 or 3 weeks behrre nor­ mal Irost dale. Less effective control may result during hot, dry sveolher when deep soil moisture is deficient. Power equipment with pressures up lo 250 pounds will aid en to.. CD /? 5 7 Gallon of Powerful, Low Volatilty Esters of 2,1,5-T • 2,4,5-T Acid Equivnl CO N TR O L OF M AN Y SPECIES OF TREES, BRUSH AND BROAD-LEAVED W In o b ta in in g sa tisfa c to ry sp ra y c o v e ra g e . R ep eo t a p p lic a tio n s m a y be n e c e ssa ry os n ew g ro w th d e ve lo p s, b ut a sin g le tre a tm e n t in a n y one yeo r is u su o lly s u ffic ie n t. NOTE: M a n y b ro o d -le a v e d w e e d s o re co n tro lle d b y th is a p p lic a tio n . STUM P T R E A T M E N T : W here qrowlh is more thon 6 to 8 lert toll, cut it clme the ground ond spray the slumps and stubs with -I gallons of Esteron 215 in 96 g:l'< ns tor I pint In 3 qoll-nst of diesel oil, fuel oil or kerosene, mixed llmtoughly. thoroughly all exposed bark, as well os cut surfaces. This means spraying until run­ down or run-off la the ground line is noticeable. Old or rough hark requires mote spray volume than young or smooth bark. Apply at any lime, including t h e w i n t e r months, prcfcrobly to ficshly-cut stumps. Best results ore usually obtained on stumps two inches across or larger. '•F R ILL ” T R E A T M E N T : For large trees, make a single-hack girdle or " frill” cf ^.ot. lopping axe cuts completely around the Iree as close lo the ground ns possible. T >*-1 1 the injured area with a mixture of 2 gollons of Eslcton 2-15 in 100 gallons I 1; pint in 3 gallons) of diesel oil, fuel oil or kerosene. A IR FL A N E A P P LIC A T IO N S: To control mesqulte, use I to I 1 1 pints o f Esteron 2 15 per ocre In 3 gollons of woter ond 1 gollon of w atcr-clcor diesel ht-l. Apply 50 to 80 dttys o iler lirsl leaves appeor. Do not treat If tlrculh hos prevented heavy frli-g c growth. For post oak and btock|ack oak, use 2 quarts of Esteron 2-15 per acre. M is this either with 3 to ‘I gollons of wolcr and I gollon of dtescl cil. or with 3 la d gallons of diesel oil. Apply oiler foliaqc Is fully developed. N O TE: Soil mnislu-c must be odequale for normol growth. Rc-trcot os accessory In succeeding years. 9 S 6 T 2 0 N11 9Z0GCH r m I Cl o BASAL BARK TREATM EN TS Brush and small trees con he controlled by spraying the basal parts of brush stems and tree trunks to a height of 12 to 15 inches from the ground line. Use a solution of 4 gollons of Esteron 2-15 in 96 gallons I I pint in 3 gallons! of diesel oil, fuel oil or kerosene. W ith certain species, 2 qollons of Esteron 2d5 In 98 gollons of diesel oil, fuel oil or kerosene is effective. Knapsack or power equipment moy be used, but complete wetting of the indicated orco is necessary, par­ ticularly at the ground line. This means spraying until run-dawn or run-off to the ground line Is noticeable. Old or rough bark requires mare sproy volume than young or smooth bark. Low pressures are deslroble. Apply ol any time, including the winter months. Often dcloyed response and killing can be expected. Treated brush or tree-, preferably should not be cut for o period of one year following application. Do ■ vr ■ LV•t |Im:' te n , vci DIn ! ,1,.. r* • in p l- w ■'1• L-rr i ■ I1 c,p pt é rmw ■s •OvU. I. *.■* + q|UJ,i ¿',. ’’., *V.* ?*‘n^V’v-’v* .?*J',,V»t:' v{[•• •'. '• . *>: »I-1■. •):.' , 1\■• wjoa.n0., «.A iiA / k r b ' t k i d U J u , 4/i.' / ',.' 'V !t‘ il. ■irftwJl. J ¿*'••> i 7»S M08SSN MULTl-USl □ » U S » A ? ! » WSSSS) » a m » »lotilty' Esters of 2,4,5-T • 2,4,5-T Acid Equivalent 4 Pounds per Gallon ¡>n I* 7 E C O N TR O L QF M A N Y SPECIES OF TREES, BRUSH A N D BRO A D -LEAVED W EEDS in oblolnino satisfactory spray coverage. Repeal applications may be necessary as new growth develops, but a single treatment In ony one year is usuolly sufficient, N O T E: Mony broad-leaved weeds are controlled by this application. BA SAL BARK T R E A T M EN T : Brush and small trees can be controlled by spraying the basal ports of brush stems and tree trunks to a height of 12 la 15 Inches from the ground line. Use a solution of -t gallons of Esteron 2-15 In 9 6 gallons 11 pint In 3 gallons I of diesel oil, fuel oil or kerosene. W ith certain species, 2 gallons of Esteron 2-15 in 98 gallons of diesel oil, fuel oil or kerosene is effective. Knopsack or power equipment may be used, but complete wetting of the Indicated area Is necessary, par­ ticularly at |he ground line. This meant spraying until run-dawn or run-aff to the ground line It goliceabte. Old or rough bark requires mare spray volume than young or smooth berk. Low pressures are desirable. Apply at any time, Including the winter m -o lli'. Often delayed response and killing can be expected. Treated brush or trees preh-'ddy should not be cut for o period of one yeor following application. STUM P T R E A T M EN T : Where growth is more than 6 to 8 feet tall, cut it close to the ground and spray the stumps ond stubs with -1 gollons cf Esteron 2-15 in 96 gollons . _ tor I pint in 3 notionsi of diesel oil, fuel oil or kerosene, mixed thoroughly. W et . jlt'or^ugbly ql| exposed bark, as well as cut surfaces. This means spraying until run'• 'd o w n ar run-aff to the ground line Is noticeable. Old or rough bark requires mare spray volume than young ar Smooth bark. Apply at any lime, including the winter months, preferably to freshly-cut stumps. Best results ore usuolly obtained on stumps two inches dcross or larger. " F R IL L " T R E A T M EN T : For large trees, make a single-hack girdle or "frill" of over­ lapping axe cuts completely around the tree as close to the ground as possible. Treol the ln|urcd orro with a mixture of 2 gollons of Esteron 2-15 in ICO gollons ( * i pint In 3 gallons I of diesel oil, fuel oil or kerosene. A IR PLA N E A P P LIC A T IO N S: To control metquite, use I to I ! i pints of Esteron 245 per acre in 3 gallons of woter and I gallon of water-clear diesel fuel. Apply 50 to 80 doys after first leaves appear. Do not treot If drculh has prevented heovy foliage growth. For post oak and blackjack oofc, use 2 quarts of Esteron 245 per acre. Mix ihls cither with 3 la 4 gallons ol water ond I gallon of diesel oil, or with 3 to. 4 gallons ol diesel oil. Apply alter foliage Is fully developed. N O TE: Soil moisture must be adequate for normal growth. R c-lreal os necessary in succeeding ycors. I x j *•l4 J W A R N IN G ! ..'it L to L come . Do not apply Esteron 245 directly to, or otherwise permit into contact svith vegetables, flowers, grapes, fruit trees, ornomcntols, cotton or other desirable plants which are sensitive to 2 ,4,5-T , ond do not permit spray mists containing it to drift onto them, since even minute quantities of the spray may cause severe injury during bath growing and dormant periods. (Coarse sprays are less likely to d rill, t Accordingly, applications by airplane, ground rigs and hand dispensers should be carried out only when there Is no hasard from drift. Do not apply by airplane in the vicinity of cot­ ton, grapes ar other desirable 2 ,4 ,5 -T susceptible vegetation. At higher temperatures vaporization may cause in|ury to susceptible plants growing nearby. Do not use on lawns of creeping grosses, such as bent, except for spot spraying, nor on freshly seeded turf until grass hos become well established. (Most legumes are usuolly damaged or killed.l Do not contaminate Irrigation ditches or water used for domest'C purposes. Do not store near fertilizers, seeds, insecticides or fungicides. To avoid injury t4 desirable plants, do not store, handle or opply other agricultural chemicals with Ihti same containers or equipment used with Esteron 24 5. Local conditions moy affect the use of herbicides. Stole agricultural authorities in many stales issue to immandations to fit local conditions. Be sure that use ol this product conforms to all applicable regulations. C A U TIO N ! M AY CAU SE SK IN IR R IT A T IO N Avoid Contact with Eyes, Skin and Clothing N O T IC E Seller makes no worronty of ony kind, express or implied, concerning the use of ibis product.. Buyer assumes all risk of use or hondling, whether In accordance with direc­ tions or not. U. S. Patents No. 2.390.941; 2 ,3 9 6 .5 1 3 ; 2,4 7 2 ,3 4 7 and 2,562,855 THE DOW CHEMICAl COr.UWflY M ID LA N D , M IC H IG A N *• * M ID LA N D D IV ISIO N 5247 966120NH (7nnPTI fJjflfl 3 * 5248 increased number of disk ings between spraying and seed­ ing resulted in a slightly lower stand of trefoil plants. This was not apparent at the Southeastern Ohio test. In other irds, a lesser amount of tillage on the 7-day interval ._rt a firmer seedbed which meant less coverage of seeds and earlier germination. Current tests in Ohio are directed at determining the minimum amount of tillage necessary with herbicide treatment. TABLE II P ercent Stand o f K entucky Bluegrass in T illed and U n -tillbd Areas R em a in in g in th e Fall o f 1936 Following H erbicide T reatments Made on __________ October 20, 1955 and April 19, 1936_________ Harbicida Rnta lb*/A Acid £ f manient fa ll Applied_______ Spring Applied Tilinga N o*tilinga Tilinga lio*tilinga D alap o n -1 0 ________ Trace Dalapon- 5 _______ Dalapon - 2 0 _______ Trace Dalapon - 1 0 _______ ATA- 4 ____________ Trace ATA- 2 ____________ ATA - 8 ____________ Trace ATA- 4 ________________ 75% Trace 45% Trace Trace Trace 30% Trace 10% 35% 90% 60% Table II summarizes data taken in a field test at the U.S.D.A. Soil and Water Conservation Research Station at Coshocton, Ohio. Duplicate plots were laid out in the fall of 1953 for spraying to compare fall and spring treatments. Estimates of stand of bluegrass in the 1955 fall and 1956 spring treated areas were made in the fall of 1956. The main point brought out in these data is the effect of tillage on percent of bluegrass kill, both fall and spring. The tillage implement used in this case was a disk harrow. All tillage plots were disked thoroughly 5-10 days after spray­ ing and fall sprayed plots were disked again in the spring prior to seeding. The bluegrass kill was almost complete on fall and spring sprayed tillage plots. The heavier fall herbicide rates without any tillage also resulted in almost complete kill of bluegrass. The importance of tillage in conjunction with spring applied chemicals is quite appar­ ent even at heavier Tates. The stand of trefoil obtained by band seeding in this test was variable and inconclusive but indications are that satisfactory stands of trefoil can be established with fall applied herbicide and no cultivation. On these fall-treated, no-tillage plots the trefoil stand was estimated to be from 50 to 80 percent. Again, winter survival will affect appraisals of results. In conclusion these data show the possibilities of using herbicides in seedbed preparation for birdsfoot trefoil. Proper use of available materials will suppress competing vegetation sufficiently to permit better stands of trefoil with a minimum of cultivation. Dalapon can be used effectively as either a spring or fall treatment It lends itself readily to spring application because of its low toxicity to birdsfoot trefoil seedlings. On the other hand ATA is probably best applied only in the fall if spring seeding is intended. There is need for further research to determine: (1) The minimum amount of tillage necessary, (2) The usefulness of the new pasture renovation seeding equip­ ment that is on the market, (3) Minimum amount of chemical to use under various conditions. REFERENCES (1) Sprague, M. A., The substitution of chemicals for tillage in pasture renovation, Agron. Jour. 44:405-409, 1952. (2) Kates, Allan H., M. A. Sprague and R. J. Aldrich, Promis­ ing chemicals for seedbed preparation, Down To Earth, Vol. 10, No. 4, 1955 (Pages 10 and 11). R ESID U E TO LERA N CES FOR DOW A G R IC U LTU R A L CHEM ICALS By G. E. Ly n n *, The Dow Chemical Company It is now well recognized that pesticidal chemicals must be used in agriculture to prevent crop and animal losses during production and storage. Pesticidal chemicals, such as insecticides and weed killers, have become a necessary part of the farming economy and take their place in importance along with the tractor and other modern farm­ ing tools. Pesticides can be used safely and without hazard to the consumer of the food crop. Although some pesti­ cides may leave trace residues on food crops, it has been shown by extensive toxicological investigations that the small amounts of most pesticides that appear in food can be consumed without any adverse effect on the health of man or animals. Under these circumstances the Miller Amendment to the Federal Food, Drug and Cosmetic Act provides a means by which adequate residue toler­ ances on raw agricultural commodities can be established to permit the use of the pesticides. The Dow Chemical Company has obtained the necessary data to establish many such tolerances for its agricultural chemical products. Additional data are being obtained to expand the list of commodities and crops on which both old and new pesticides may be used. Many pesticidal chemicals are used in ways which do not result in a residue. Application early in the growing season, or use at low dosage rates often eliminates the residue problem. Numerical tolerances are not necessary under these circumstances. These chemicals are, however, subject to registration and ample proof of the lack of resi­ due must be submitted to the United States Department of Agriculture before a registration is granted. For example, Premergef, containing dinitro-o-sec butylphenol, is recom­ mended for a variety of pre- and postemergence herbicidal uses. Each use has been investigated thoroughly not only for effectiveness on weeds and safety on crops but also for possible crop residue. When used as directed on the label, there is no crop residue problem. Users of agricultural chemicals are urged to follow the directions that appear on the labels. Any other use, or deoarture from label (Continued next page) 'Midland, Michigan. t Premerge is a trademark of The Dow Chemical Company. 5249 Dow n t o \E a r t h , Spring, 195 7 . i Chemical Company has carefully investigated these chem cals and when each is employed in accordance with got agricultural practice, as directed on the label, no pubi health hazard will result. The following information summarizes the status many registered Dow Agricultural Chemical products as January 15, 1957. recommendations, may result in poor performance and residue problems not covered by tolerances. Some chemicals, used in agriculture, are not classified as pesticides by the Federal Law and thus cannot be regis­ tered. Dow agricultural chemicals in this category are Magron, Colorset, App-l-set, anhydrous ammonia and the soil fumigants, Fumazone, Telone and Dorlone. The Dow R ES ID U E T O L E R A N C E STATUS FUMIGANTS D ow A gricultural Product A ctive Ingredients Dowfume * EB-5 and Dowfume EB-15 ethylene dibromide ethylene dichloride carbon tetrachloride Dowfume 75 ethylene dichloride carbon tetrachloride Dowfume 80-20 carbon tetrachloride carbon disulfide Dowfume C carbon tetrachloride carbon disulfide ' ethylene dibromide Dowfume 1; carbon tetrachloride ethylene dichloride ethylene dibromide sulfur dioxide Dowfume G ethylene dichloride carbon tetrachloride ethylene dibromide Tolerances Carbon tetrachloride, ethylene dichloride, carbon disulfide, and the orga bromide residues from ethylene dibromide are exempted from the requ ment of a tolerance when used as a fumigant for the following grains: barley, corn, oats, popcorn, rice, rye, sorghum (milo) and wheat. Tolerances of 50 ppm are established for residues of inorganic bromi (calculated as Br) in or on the following grains that have been fumiga with ethylene dibromide: barley, corn, oats, popcorn, rice, rye, sorghum (milo) and wheat. Sulfur dioxide in amounts used does not constitute a residue problem. Dowfume H carbon tetrachloride carbon disulfide sulfur dioxide Dowfume J ethylene dichloride carbon tetrachloride ethylene dibromide Verri fume* carbon tetrachloride carbon disulfide Serafume* carbon tetrachloride ethylene dibromide ethylene dichloride carbon disulfide Methyl Bromide methyl bromide The tolerances for residues of inorganic bromides (calculated as Br) or on raw agricultural commodities which have been fumigated with met bromide are as follows: (a) 5 ppm in or on apples, pears, quinces. (b) 20 ppm in or on eggplants, onions, tomatoes. (c) 30 ppm in or on beets, rutabagas, turnips. (d) 50 ppm in or on alfalfa, hay, barley, brans, green brans, lima brans, snap beans, corn, grain sorghum (milo), oats, rice, rye, wheat. (e) 75 ppm in or on potatoes, sweet potatoes. (f) 200 ppm in or on cottonseed. Dowfume MC-2 methyl bromide chloropicrin When used in accordance with directions as a plant bed treatment and w' transplants are moved to soil not treated with this fumigant, no detear residues are probable in or on a food crop. Dowfume W-85 ethylene dibromide The tolerance for residues of inorganic bromide (calculated as Br) in or raw agricultural commodities grown on soil treated with ethylene dibrorr are as follows: (a) 5 ppm in or on lima brans, strawberries. (b) 10 ppm in or on asparagus, cauliflower. (c) 25 ppm in or on cotton. (d) 50 ppm in or on sweet potatoes. (e) 75 ppm in or on carrots (with or without tops), parsnios. Picfume* chloropicrin Leaves no residues on food crops when used as a soil fumigant in accord: with Dow label instruaions. Trademarks of The Dow Chemical Company 10 5250 * % HERBICIDES rt'tw A gricultural P roduct A ctive Ingredient! Tolerances Dowpon* -dalapon, sodium salt (2,2-dichloropropionic add, sodium salt) Tolerances established for 5 ppm in or on sugar bens and 35 ppm in or on cottonseed (calculated as 2,2-dichloropropionic add). No residues result from early pre-planting treatments when used in accordance with Dow label instructions. No residues result from use on sugar cane. Dow Sodium TCA 90% trichloroacetic add, sodium salt dinitro-o-sec butylphenol, alkanolamine salts dinitro-o-sec-butylphenol, ammonium salt dinitro-o-sec-butyiphenoi, When used in accordance with Dow label recommendations, no residue in or on a food crop will result. Premerge* Dow Selective Weed Killer Dow General Weed Killer tmeron* 44 tirteron Ten Ten 4Estexon 99 Esreron 76E Ccteron Brush Killer Csteron 245 Reddon* flow 2,4,5-T Amine :ed Killer i\.uron* 2,4-Dow Weed Killer, formula 40 Home Use Weed Killer Dow MCP Amine Weed Killer Zero tolerances established. When used in accordance with Dow label recommendations, no residues in or on a food crop will result. 2,4-dichlorophenoxyacedc add, isopropyl ester 2,4-dichlorophenoxyacetic add, propylene glycol butyl ether esters 2.4-dichlorophenoxyacetic add isopropyl ester and 2.4-dichlorophenoxyacetic acid, butvl esters 2,4-dichlorophenoxyacetic add, propylene glycol butyl ether esters, and 2,4,5-trichlorophenoxyacedc add, propylene glycol butyl ether esters 2,4,5-trichlorophenoxyacetic add, propylene glycol butyl ether esters 2,4,5-trichlorophenoxyacetic add. triethylamine salt 2 (2,4,5-trichlorophenoxy ) propionic add, propylene glycol butyl ether esters 2,4-dichlorophenoxyacetic acid, alkanolamine salts When used in accordance with Dow label recommendations, no residue in or on a food crop will resu lt (Note: A tolerance of 5 ppm 2,4-dichlorophenoxyacetic acid has been established in or on apples, dtrus fruits, pears, and quinces.) — 2-methyl-4-chlorophenoxyacetic add. alkanolamine salts * Trademarks of The Dow Chemicai Compûnj INSECTICIDES D ow A gricultural Product A ctive Ingredients Tolerancei DN-Dry Mix* No. 1 dinitro-o-cydohexylphenol A residue tolerance of 1 ppm is established in or on citrus fruits. DN-11I* dinitro-o-cydohexyiphenol, dicydohexyl amine salt A residue tolerance of 1 ppm is established in or on apples, apricots, beans, blackberries, black-eyed peas, celery, cherries, citrus fruits, grapes, logan­ berries, nectarines, peaches, pears, plums, (fresh prunes), quince, and raspberries. DN-Dry Mix* No. 2 dinitro-0-cresol Zero tolerance established. When used in accordance with Dow label recom­ mendations, no residue in or on a food crop will result. DN-289* dinitro-o-sec. butylphenol, tri-ethanolamine salt Zero tolerance established. When used in accordance with Dow label recommendations, no residue in or on a food crop will result. Ovotran* Wettable ovex (p-chlorophenyl p-chlorobenzenesulfonate ) Residue tolerances are established as follows: (a) 3 ppm in or on apples, peaches, pears, and plums (prunes). (b) 5 ppm in or on grapefruit, lemons, oranges, and tangerines. Dowicide* A Sodium-o-phenylphenate A tolerance of 10 ppm is established for residues of sodium o-phenylphenate (calculated as o-phenylphenol) in or on each of the following citrus fruits: oranges, lemons, limes, grapefruit, tangerines, rangelos, citrus citron, kumquar. A tolerance of 5 ppm is established for residues of sodium o-phenylphenate (calculated as o-phenylphenol), in or on apples and pears. * Trademarks of The Dow Chemical Company ii 5251 D o w n to Ea r th , Spring, 1957 5252 A CD file no. HG-51 V T O X I C I T Y OF "HERBICIDES TO DOMESTIC ANIMALS by T. A. Hymas, DVM Agricultural Chemical Research THE D O W C H E MICAL COMPANY Mid l a n d , Michigan May 29, 1958 I n reviewing the p a p e r that I a m about to present I believe the title should more appropriately be "The Lack of Toxicity of Herbicides to Domestic Animals" rather than "Toxicity of Herbicides to Domestic Animals." T h e use of herbicides f o r the control of undesirable vegetation has been practiced f o r many years, some of the older materials that have been used a r e the chlorates, arsenlcals, boron compounds, certain oils, and even table salt. While some of these materials are still being used on occasion f o r this purpose, r e s earch over the past 14 years has revealed new h e r b i ­ cide materials that are superior to the older materials and in some instances a re less toxic to animals and also man. The herbicides that I wish to d i s ­ cuss at this time are organic chemical compounds that act 3 ystemically when applied to vegetation. T hey have a selective toxicity to specific liants and a r e currently being used throughout the world f o r the control undesirabl e vegetation. I am going to confine my remarks to several of the 2,4-D type materials which can be classed as chlorinated phenoxy-acetic acid compounds, and to a relatively n e w comer to the field of selective herbicides, dalapon, known chemically as 2,2-dichloropropionic acid. T o begin, I would like to give some comparative data that has been ob­ tained by numerous investigators, to compare the toxicity of these herbicides when gi v e n orally to different species of animals. In Table No. 1 the ac u t e oral L D c q values for several species of animals fed the most widely used chloropnenoxy-acetic acid compounds is given. These are 2 , 4 - D (2,4-dlchlorophenoxyacetic acid), 2,4,5-T (2,4,5-trichlorophenoxy-aceti c acid), and silvex which is 2-(2, 4 , 5 - t r i c h l o r o p h e n ox y) propionic acid. The chlorinated phenoxyacetic and propionic acid compounds are very s i m i l a r in chemical make-up, they are also similar in their toxicity to animals as one might expect. Dogs a p pear to be the most s u s ­ ceptible species to the chlorinated phenoxy acetic and propionic acid materials. Where there are two sets of figures given in the table they represent a range of values to cover the pure acids, also esters and salts of the compounds, sex differences, and work by different investigators. While, the basic acid has not b e e n greatly altered when formulated, as an ester or salt, the foiroulation may only slightly alter the biological ctivity of the toxicant as measured by herbicidal activity and toxicity ico animals. 5 2 5 3 The value giv e n for cattle was obtained using a mixture of 2,4-D and r% it c m 2,4,5-T. 0003361 Tl ’H -2 - Becauae of the similarity of toxicity of 2,4-D and 2,4,5-T to laboratory animals and the wide use of the combination of these herbicides in . commercial products, it was decided to use a combination of the two materials (Esteron* B r u s h Killer) containing approximately equal parts of 2,4-D and 2,4,5-T as esters, to study the acute oral toxicity of the materials in cattle. t The cattle used for these studies were dairy cattle of mixed breeding weighing fr o m 250 to 336 kilograms. Table No. 2 is a summary of the work accomplished. Experiment # 1 : A steer weighing 291 Kg. was given a single oral dose of 1,000 mg/Kg acid equivalent of E s t e r o n B r u s h Killer, sufficient herbicide to prepare 1 1 gallons of spray at the highest level recommended, 4 qts /100 gal. of spray, or 1 6 .6 gallons at the usual recommended rate of 3 qts/100 gal. of spray. This dosage had no observable effect on the steer. l A second st e e r weighing 295 Kg. was given 1,000 mg/Kg acid equivalent of this same mixture of 2,4-D and 2,4,5-T, as E s t e r o n B r u s h Killer, on each of three successive days. The quantity of herbicide given would treat 1/2 acre of heavy brushy, woody, or weedy area at the recommended use level. T he day f ollowing the third dose a general depression was noted, the animal was off feed and lethargic. The general attitude and behavior became progressively worse until the animal died on the third day following the last dose. D e a t h was undramatlc. The steer evidenced no pain, no struggling, or signs of violence. Upo n autopsy it was noted that the rumen had a strong odor that is characteristic of the 2,4-D type materials. The reticulum, rumen, omasum, a b o m a s u m and fore-part of the duodenum were abnormally dry and Impacted. R u m e n stasis was evident. Beyond the duodenum the G. I. tract contents were fluid. No other gross abnormalities were evident. A third st e e r weighing 295 Kg. was given 500 mg/Kg on each of two c o n secu­ tive days. Oh the third day the animal was off feed and rumen motility had essentially ceased. Otherwise the animal appeared normal. On the fourth day, this steer appeared normal and there were no discernable af t e r effects. A f o u r t h animal weighing 336 Kg. was given 500 mg/Kg on e a c h of three successive days and no toxic symptoms were observed. The quantity of herbicide g i v e n would make 2 7 -5 gallons of spray at the recommended con. centration of 4 lbs. acid equivalent per 100 gallons of spray. The animal remained on full feed without showing effects of a n y kind. < A fifth st e e r weighing 2 5 0 Kg. was then given 100 mg/Kg of the 2,4-D, 2,4,5-T mix t u r e on 15 consecutive days and observed carefully. There were no outward appearances of adverse effects. The animal was necropsled 48 hours following the last dosage. Gross examination of the internal organs revealed only slight petechial hemorrhage in the d u o denum and a very mild diffuse irritation in the abomasum. Histological examination of U tissues revealed very mild pathology in the liver and kidneys. The liver contained small areas of focol hemorrhagic necrosis surrounded by areas 5254 * Trademark of The Dow Chemical Company 0003362 I' of f a t t y degeneration. In the kidneys a slight interstitial adema and congestion of the cortical medullary regions was observed. The a m o u n t of 2 , 4 - D and 2,4,5-T that was ingested by this steer was 0.8 lb., wh i c h is sufficient to make 20.5 gallons of spray to treat dense or hard to kill fol i a g e or herbage. It would be Impossible for one or more bovine to consume all of the toxi- c cant applied in 2 0 to 30 gallons of spray even if they were confined in c an area where all vegetation was treated. Some toxicant drips or is c sprayed on the ground and m uch vegetation that is sprayed is of an unpalatable nature being brush, weeds or the like. I do not believe the chlorinated phe n o x y acetic acid herbicides constitute any hazard whatsoeve: to cattle If they are handled with reasonable care and used according to recommendations of the manufacturer. I would n o w like to consider dalapon few minutes. ^ j (2,2-dichloropropionic acid) f o r a Slide # 3 gi v e s the a c u t e oral toxicities of dalapon In several species of animals. In eva l u a t i n g these figures it is well to r e m ember that the LD c q value f o r s o d i u m chloride (table salt) is 2,500 to 3,500 mg/Kg In laboratory animals. The figure reported f o r dogs, an L D c q of greater than 1,000 mg/Kg, is the largest dosage that dogs would retain without emesis. We have found that quite often it is very difficult to determine acute oral toxicities in dogs because of their ability to regurgitate such materials and our inability to control this reflex even with the use of morphine, various sedatives, tranquilizers, etc., which, of course, we are always reluctant to use because of the effect of the anti-emetic Itself. I would like to say at this point that where we have had an opportunity to compare the ac u t e oral toxicity values of herbicide materials In small animals w i t h those obtained using cattle, the two values are usually close. If real differences exist, to date, in our work, cattle have tolerated the l a rger doses, perhaps because of the bact e r i a l flora in the rumen and t h e i r a b i l i t y to deco m p o s e organic materials. \.y We attempt e d to a t t a i n acute oral L D e Q values f o r dal a p o n In cattle but, as can be s e e n in slide #4, w h e n 4,000 mg/Kg, approximately 3*5 lbs. of the chemical was "pumped" into a steer at one time, 5-5 lbs. was "pumped" into a n o t h e r steer ove r a ten day period and a n equal amount on a weight basis "pumped" into a suckling calf, and the worst reaction seen was a transient d i g e s t i v e upset and, I might add, a very effective grass killing Job in o u r grass pasture where u rination occurred following dosing. We gave up t r y i n g to kill cattle with d alapon as it appeared they would need to be drow n e d in it. Histopathological studies of tissues f r o m the animals that were sacrificed revealed no abnormalities. We concluded that dalapon was not a hazard on farms, rights-of-way, in ditches, or far m ponds, or under a n y conditions of practical use. This conclusion has b een substantiated by several years use in this country and o t h e r countries without a single substantiated case of poisoning being brought to our attention. And, we believe if poisoning cases had occurred we would know about them. B e i n g associated with a chemical company f o r several y&lHl336 r ' 5255 - 4- I know that where chemicals are used and any abnormality is observed, the first.thoug h t is u s u a l l y — the chemical did it. Any legitimate chemical manufacturer s has always expended considerable time and effort to deter­ mine the hazards of having chemicals used where men and animals are likelyrto be exposed. £2 A great deal of work and effort always goes into toxicology work on the ^ product a l s o the preparation of labels and literature to inform the consumlng public of the limitations, handling hazards and possible hazards ^ that may be encountered when the chemical is used. ^ D a l a p o n is being widely used to control undesirable grass species such a s ^ J o h n s o n g r a s s , quackgrass, and is also being used to control the d e s l r a b l e Q species of grass that are growing in undesirable locations. It is also finding wide usage to control cattails and marsh grasses that so commonly choke drain a g e ditches, waterways, farm ponds, wildlife refuges, etc. The a ctive material may be applied at a rate of f rom 1.5 to 40 lbs. per acre d e pending on the type of problem being attacked. When the material is prepared as a spray up to 20 lbs. p e r 100 gallons, a 1,000# cow would have to drink at least 15 gallons of the undiluted most concentrated spray mix to evidence the slightest symptoms of intoxication. And she could probably survive eve n a dose of double this quantity or 30 gallons of the 20 lbs', per 100 g a l l o n spray mix. If a 1,000# cow had access to a spray tank c o n ­ taining the usual and most frequent concentration of dalapon, 10-15 lbs. p e r 100 gallons, she would need to drink 33 bo 50 gallons of the spray mix to evidence the slightest intoxication. It Is interesting, to n o t e that since the advent of the 2 , 4 - D herbicides, over 225 mil l i o n lbs. of the herbicide have been used. This would be sufficient to treat some 565 million acres of land at a practical use level of 0.4 lbs. acid equivalent p e r acre. To date, to our knowledge, there has not be e n one single substantiated case of poisoning due to the herbicide. As you g e n t l e m e n might guess, there have been numerous a l l e g a ­ tions and there have b e e n a n u m b e r of cases of reported poisonings that have occurred f ollowing appli c a t i o n of herbicides, but in every case that has b e e n followed up and a diagnosis was possible, the herbicide has been exonerated. There has b e e n m u c h d i s c ussion of the possibility that spraying with herbicides might cause some plants to become toxic and other plants that are ordinarily non-palatable to become palatable. Considerable investiga­ tion of these possibilities has resulted in the general conclusion th^t from a pract i c a l standpoint these suppositions are more fanciful than factual. To date the only effect on herbage that has bee n proven that might be harmful to livestock is in the case of certain plants that have been sprayed with 2,4-D or s imilar materials. The nitrate content can be i n ­ creased. F o r instance, in the case of sugar beets, accidentally sprayed with 2,4-D, the nitrate content of the leaves is actually Increased and it is possible that the feeding of sugar beet tops treated with 2,4-D could cause nitrat e poisoning. Of course, the only time 2,4-D would be applied to sugar beets is by accident, as the material is not recommended for use in sugar beet production and has never been applied for thinning purposes. It has b e e n reported that other plants, such as l a m b s q u a r t e r s , pigweed, ' 5256 0003364 4 C c. t I »OPS om a nd smartweed will increase their nitrate content a f t e r treatment with 2,4-D. Since livestock do not ordinarily eat these species, it would not s e e m to be a praqtical problem, although it is possible that if animals were starved to eating suc h plants and the plants were sprayed with 2,4-D that nitrate poisoning could occur. It might be pointed out that environ­ m e n t a l and mechanical factors can cause as significant changes in nitrate level as herbicides. T h e ques t i o n has bee n raised many times: Will wild cherry present a special problem whe n sprayed w ith 2,4-D type herbicides? It has been r a t h e r conclusively p r o v e n by Grigsby and Baugh, also G rigsby and Blakeslee at the Michig a n State University, also by Bar r o n s and Lyn n of The Dow C h e m i c a Company that wild cherry does not increase in H C N content when it is treated with 2,4-D. Thi s has been proven by chemical methods and further substantiated by practical type tests. In a test conducted by Grigsby and Bla k e s l e e of the M i c h i g a n State University and L. L. C o u l t e r of The Dow . Chemical Company, sheep were pastured on wild cherry that had b e e n sprayed. Immediately pr i o r to the time the sheep were turned into the brushy pasture, two quartersectlons of the pasture including the wild cherry plants were sprayed with a mixture of 2,4-D and 2,4,5-T, the sheep graced the wild cherry p l a n t s — not only at the time they were turned into the pasture but f o r several days following, and until the vegetation actually turned yellow and brown, and was so m a r k e d l y affected by the herbicide that it was no -^longer palatable. At no time was there any adverse effects noted in these Jsheep, and at n o time was there evidence that the treatment of the herbage affected the normal habits or the normal functions of the sheep that grazed on It. Th e r e was no preference shown f o r either treated, or untreated plants. I n all of the alleged cases of herbicldal p oisoning that have b een definitely diagnosed, the diagnosis has b e e n such causes as: shipping fever, leptospirosis, a n a p l a s m o s i s , arsenic, poisonous plants, lead, p o o r ma r k s ­ m a n ship (hot lead), parasites, hardware disease, etc. It should be born in mind that many types of weeds and foliage have in the past and will continue to cause poi s o n i n g w h e t h e r they are sprayed or whe t h e r they are not sprayed wi t h herbicides. It is a l s o known that various members of the;‘sudan grass and sorghum f a m i l y have caused difficulties in the past and undoubtedly will cause them in the future. Here, again, losses occurred p r i o r to the use of m o d e r n herbicides a n d -will continue despite herbicide usage. Some workers have felt that these systemic herbicides may improve the palatablllty of n o r m a l l y unpalatable poisonous plants to a degree where animals wi l l graze them. However, in actual gra z i n g trials on desirable grasses and legumes, livestock have never shown preference f o r the treated foliage. In fact, where there has been a preference shown, it has always b e e n f o r the untreated plants, and until it c a n be proven that the palatablllty of a n y plants is actually increased by the use of herbicides, we should keep a n open mind. I have prepared a table (Slide #5) to show the m i n i m u m and max i m um spray concentration of the products used and the min i m u m quantities of spray a ‘-'1,000# cow would need to drink, or sprayed area she would have to consume all of the vegetation f rom (assuming all spray is retained on the vegetation) to evidence even mild intoxication f rom ingestion of the herbicide. 5257 000336 -6- Y o u should remember that the more concentrated the spray, the heavier and more u npala t a b l e the vegetation to be s p r a y e d . I have brought along a few slides to give you a better Idea of Just how these materials work and where you might expect to see them used. £ \ 9 n p q ni11 t c Table #1 ACUTE ORAL TOXICITIES OF 2,4-D a n d r e l a t e d c o m p o u n d s LD50 (MsAg) 2,4-D 2,4,5-T Sllvex Mice 375-713 389-551 1410 Rats 375-805 481-500 600-621 Rabbits 424-800 712 750-819 Guinea pigs 4 6 9 -1000 381-750 1200-1250 Dogs 100 100 — Chicks 540-2 0 0 0 310 1190 Animal Species . % Cattle >1000* * Mixture of equal parts 2,4-D and 2,4,5-T 5258 000336 i * - 7 - S' Table #2 TOXICITY OP A MIXTURE OP 2,4-D, 2,4,5-T GIVEN ORALLY T O CATTLE Dosage Mg/Kg. S-291 1000 Observations No effect S-295 1000/da. f o r 3 da. Fatal S-295 500/da. for 2 da. Moderate transient toxicity S-336 500/da. f o r 3 da. No effect S-250 100/da. f o r 15 da. G rossly no effect. Microscopically, liver & kidney mildly affected. Table # 3 A C U T E ORAL TO X I C I T Y OP DALAPON (Sodium Salt) A n i m a l Species L D c jQ V a lues Mice >4,000 Rata 6 ,600- 8 ,10 0 G u i n e a pigs 3,400 Rabbits 3,400 V Dogs >1,000 C h icks 5,700 Cattle >4,000 5259 o o < m $ ? u u n u O J U d I J Sex and Wt. In Kgs. Slide #4 TOXICITY OP DAL A P O N (Sodium Salt) G I V E N ORALLY T O CATTLE Dosage MgAg_ Observations S-336 2,000. No effect S-290 3,000 Mild transient toxicity S-337 3,500 Mild transient toxicity S-394 4,000 Mild transient toxicity P*-252 1000/da. f o r 10 da. M o d erate toxic symptoms S*-59 1000/da. f o r 10 da. N o effect * 180£9QM0(I S e x and Wt. in K g s . * Necropsied f o r h i s t o p athology studies Slide #5 Herbici d e 2, 4 - D and 2,4,5- T Dalapon (Sodium salt) Sp r a y C o n ­ centration lbs/100 gal. 1000# Cow May B e c o m e IntoxIcated if She Ingests Spray Con- . All Spray from Sprayed Area centrate (Sa. feet) (Gallons) 0.25 400 174,240 4.0 25 10,890 1.5 333 143,748 20.0 25 5,445 5260 0003368 3 1 52S1 3T F O R E S T R Y LIBRARY JU N 2 7 í: . ú UNIVERSITY OF CALIFORNIA BERKELEY eing more susceptible to the than Scotch (P. sylvestris t red (P- resinosa Ait.) pine, though environmental condicannot be rigidly controlled e field during and after aptions, spraying should be done er optimum conditions of teni­ a e and light intensity to in­ minimum seedling damage. servation Dept., Albany, N. Y. 3. F aulkner , B. 1952. Notes on nursery irrigation and on chemical weed con­ trol practices in the U. S. A. and Canada. Forestry 25(2): 126-134. 4. L aCroix , J. D., and A. T. Guard. 1956. Morphological and histological modifications of pine seedlings in­ duced by petroleum naphtha. Canad. Jour. Botany 34(4) : 621-627. 5. S toeckzler, J. H. 1949. Control of weeds in conifer nurseries by mineral spirits. Lake States Forest Expt. Sta. Paper No. 17. 5 percent of the sprayed trees sprouted. Relative effectiveness of the treatments as site preparations is revealed by survival of planted loblolly pine seedlings. In Novem­ ber 1954, after a very dry summer, loblolly survival was 53 percent on the plots where silvicide was ap­ plied on stumps, 55 percent where stumps were left untreated, and 83 Literature Cited percent where hardwoods had re­ J. D. L aCroix an d A. T. G uard ‘ SSI t t , F. M. 1951. Use of petroleum roducts as selective herbicides in Respectively, Department of ceived basal spray. The silivicide uthern pine seedbeds. Jour. PoresBiology, University of Detroit, had no adverse effect on the pine 19: 773-775. Detroit, Mich.; Department of seedlings under either treatment, u s o n , E. J. 1949. The use of oil Lrays for the control of weeds in Biological Sciences, Purdue even when planting followed its ap­ Coniferous nurseries—1948 supple­ University, Lafayette, Ind. plication by only a few hours. ment. Mimeog. by N. Y. State Con­ However, none of the three methods afforded an adequate “ one shot” release treatment for Texas conditions. A f t e r t h r e e growing seasons, two of which were very dry, pine survival on the two cut treatments had declined to 45 and 27 percent for the areas with and without silvicide, respectively, asal Spray with 2,4,5-T for Winter Hardwood Control while on the basal-spray area it In East Texas was down to 64 percent. Many of the pines needed release—on the ■Removal of competing hard- butyl ether ester) in 16 gallons of cut areas from hardwood sprouts, "oods is essential to re-establish- No. 2 diesel oil, a concentration of and on the basal-spray areas from ent of pine on many dry sites, 23.5 pounds of acid per hundred invading herbs and vines and from erial spraying of silvicides or gallons. In both treatments the the expanded crowns of the few and-c-learing with heavy machin­ solution was applied to the point surviving hardwoods. Had adequate ery promises to be useful on ex­ of runoff. release been given at the end of the Of the original stand of 13-yeartensive operations. For owners of second year, 80 percent of the pines mail tracts, however, hand meth- old mixed hardwoods, 20 sweetgum on the basal-spray areas might ds that can be applied at planting (Liquidambar styracifi.ua L.) trees have survived, as compared with or stumps on each plot were ob­ .time are most acceptable. less than 50 percent on the other ■iSearch for such a method at the served for three years. By April treatments. ustin Experimental Forest in east 1954, 92 percent of the stumps The poor showing of 2, 4, 5-T Texas revealed that for Decem­ without silvicide had sprouted, solution on cut stumps appears to ber application, and, presumably and this increased to 100 percent be related to time of application. In throughout the dormant season, 2K by the following November. The 2, tests made during the growing sea­ 5-T is more effective when ap- 4, 5-T solution on cut stumps de­ son even more dilute solutions on lied as a basal spray than when layed but did not reduce sprout­ cut stumps have permanently in­ ing : 37 percent of the stumps painted on cut stumps. hibited sprouting. For applications ~ Triplicate sets of plots were sprouted by April, 90 percent by concurrent with planting, which treated and planted to loblolly pine November, and 97 percent by April must be done during the dormant .(Pinvs taeda L.) in December 1955. Untreated stumps produced season, 2, 4, 5-T is apparently best 1953, near the beginning of the nor­ significantly larger and slightly applied as a basal spray. mal planting season. On one set of more sprouts than stumps with plots all hardwoods were basal- silvicide, but by the third growing J ames R. D avis sprayed, on a second set all hard­ season sprouts on both treatments Southern Forest woods were cut close to the ground, had outgrown the surviving pine Experiment Station,1 a&d on the third the hardwoods seedlings. Forest Service, U. S. Were similarly cut and the stumps Ninety-three percent of the Department of Agriculture Painted with silvicide. The silvicide basal-sprayed tr e e s e v e n t u a l l y l-°r the first and third treatments died, though more than half of 'East Texas Research Center, main­ in cooperation with Stephen F. was a mixture of 1 gallon of Dow them retained their leaves through­ tained Austin State College, Nacogdoches, , steron 245 '(a propylene glycol out the 'first growing season.1' Only T““ 5263 5264 \ 31a > Krprinlrd from the J ournal o r F orestry, Vol. 57, No. C, June 105» V Concentrated or Diluted 2,4»5-T as a Supplement to Girdling? John J. Stransky T h i s a r t ic l e reports on tests in­ stalled to determine 'whether fallstrength 2,4,5-T concentrate, ap­ plied in a fine spray, would control sprouting and hasten crown kill of hardwoods girdled by ax or ma­ chine. A n earlier study evaluated the effect of 2,4,5-T on cut stumps 2). A supplementary chemical treat­ ment is needed where vigorously sprouting hardwoods are girdled to release small seedlings, or where, as in east Texas, sprout control is essential to conserve limited mois­ ture (4,5). An effective concentrate would reduce the bulk of liquid enough so that the silvicide could be carried and applied by the ax man or operator of the girdling ma­ chine, thus eliminating the heavy transportation costs and separate { T a r author is on the staff of the South­ ern Forest Experiment Station, Forest Service, 17. S. Department of Agriculture, E ast Texas Research Center, maintained in cooperation with the Stephen F . Aus­ tin S tate College, Nacogdoches, Texas. stems and were judged free of un­ derground connection with their neighbors. Girdling was either by ax, in Post oak (Quercus steUata Wanwhich case a standard double-hack genh.) and sweetgum (Liquidambar ttyracif.ua L.) were chosen as girdle was cut, or by the “ Little Beaver” power girdler, produced test species because of their preval­ by the Haynes Manufacturing ence as weed trees on upland pine sites and because both sprout pro- Company Livingston, Texas. This lifically when girdled. In some areas gasoline-powered machine employs sweetgum has also shown a ten­ a flexible shaft to power a cutting dency to produce enough callus head which produces a smooth, growth to bridge the shallow cut rounded incision about % inch wide. Properly used, it cuts made by the power girdler. through the bark and well into the The study was conducted on up­ sapwood, completely severing the land pine-hardwood sites within the cambium. Stephen F. Austin Experimental The chemical was a commercial Forest, near Nacogdoches, Texas. preparation of 2.4,5-T, Dow EsSoils are sandy loams overlying teron 245, containing 42.5 percent clay, and fairly representative of propylene glycol ether ester by the better Upper Coastal Plain sites weight (4 pounds acid equivalent in east Texas. Study trees were of per gallon). The dilute solution was two d.b.h. classes. 3.6 to 5.5 inches made up with Diesel oil, in propor­ and 5.6 to 7.5 inches. All had single tion of 1 to 50, and was painted on the trees with a brush. The concen­ trate was applied to both ax and ‘This study was designed and partially machine girdles from a plastic installed by T. A. H arrington. application required with standard dilutions. D esign 1 5265 4*»>>'■i1 J o k e 1959 ¡«squeeze bottle which dispensed a very fine spray. Both dosages were applied in such quantity as to give approximately equal amounts of %4,5-T acid to each tree. The main installations, in April 1955, subjected sweetgums and post oaks to ax and power girdling with­ out chemical, with dilute chemical, and with the concentrate. On each of the two species the six treatments were randomly applied, so that ten trees in each of the two size classes received each treatment. There were three replications of the treatment pattern. Since other workers (3, 6) have found seasonal differences in re­ sponse to silvicides, parts of the ex­ periment were replicated in Janu­ ary, July, and November 1955. Five sweetgum trees were treated per size-treatment group. Treatments were limited to power girdling with dilute, concentrated, and no chem­ ical. There were three replications each season, as in the main study. The amount of concentrate or equivalent applied in each treat­ ment is shown in Table 1. All trees were inspected one and two full growing seasons after be­ ing treated. Crown condition, num­ ber of sprouts per tree, and length of dominant sprout were criteria for determining treatment effec­ tiveness. This report is based on the second year’s data. Method of Girdling Without chemicals, ax girdling killed crowns of 90 percent of treat­ ed trees, significantly more than girdling with the machine, which killed 75 percent (Table 2). The difference was greatest in post oak; there tops of all ax-girdled trees died within two years. With chem­ icals, the two girdling methods were substantially equal, both in top-kill, which consistently exceeded 90 per­ cent, and in percent of completely dead trees, which varied with con­ centration. All tops still alive after two years were damaged, and are expected to die eventually except in the few cases where the girdle has been bridged by callus tissue, or where the tree is sustained by root grafts. T a b l e 1.— Co n c e n t r a t io n Season of installation E q u iv a l e n t P e r T r e a t m e n t oh 2,4,5-T concentrate per tree As concentrate As dilute solution Species Sweetgum Sweetgum and post oak Sweetgum Sweetgum W inter -Spring Summer Fall Ce -2.2' .9* 1.0 1.0 Ce 1.0 1.0 1.0 “Thinned 50-50 with Diesel oil to facilitate application in low temperature. This ■light dilution produced resulta comparable to the full-strength concentrate and was not distinguished in the analysis. T able 2.—Average P ercent o r Sweetoum and P ost Oak T rees Crown-Killed and Completely D ead T wo T ears A rre s T reatment in April 1955 Treatm ent Crown-killed with and without sprouts Crown-killed without sprouts Percent1 No chemical Power g ird le ----------------------------------- , 75 Ax g ir d le ___ !--------------------------------90 Mean — ------------------------------------------------- 85 Concentrated 2,4,5-T Power g ir d le _____________________________89 Ax girdle ..... ■■ ■ ------96 _______ ______________________ 99 Dilute 2,4,5-T Power girdle .---------------------------------At girdle Mean M w ii 99 100 100 40 46 42 55 57 56 87 86 _______________ 87 “Mean values weighted by are sin transformation. effective, and provided acceptable sprout control. Of the trees that remained alive below the girdle, those that received dilute acid applications developed fewest sprouts. Results with the concentrate were only a little better than those from simple girdling. Chemicals The height of the dominant sprout A ll chemical treatments gave a follows a similar pattern, averag­ highly significantly better topkill ing about 0.8 foot shorter with di­ than simple girdling. Both concen­ lute 2,4,5-T than with the other trated and dilute 2,4,5-T were ef­ two treatments. fective, killing the crowns of more The superior effectiveness of the than 97 percent of the treated trees. dilute solution suggests that the While there was little difference Diesel oil rather than the 2,4,5-T between concentrated and dilute might be the effective agent. Studies acid in top-kill efficiency, the con­ by Campbell and Peevy (1), how­ centrate proved to be a poor sprout ever, as well as informal tests in inhibitor. In the April tests, the Texas, indicate that Diesel oil alone average percentage of completely has little sprout-inhibiting quality. dead trees was not high enough for It appears rather that effective the concentrate to be significantly quantities of 2,4,5-T are translo­ better than simple girdling; in the cated into tissues below the girdle seasonal test with sweetgum the only when diluted. This parallels complete kill percentages were experience with 2,4,5-T on cut identical (39 percent) for both of stumps, where large quantities of these treatments (Fig. 1). The dilute chemical were most effective dilute solution, with a complete kill (2). Apparently the rather viscous of 87 percent on both species after concentrate is not readily absorbed spring application, and an average into the wood, while the thin Dieselof 56 percent on sweetgum in the oil solution penetrates much more seasonal test, was significantly more readily. The difference between the two gir­ dling methods, therefore, is believed to reflect promptness rather than completeness of kill. With both methods sprouting was excessive without chemicals. 5266 * 434 J o u r n a l o f F o r estry NO CROWN KILLED: WITH SPROUTS 2,4,5-T CONCENTRATED DILUTE 2,4,5-T 2,4,5-T E3 EE) WITHOUT SPROUTS F lo. L.—Crown kill and sprouting of girdled sweetgums, by season and treatm ent (average of ax and machine girdling). Season, Species, Tree Size In the sweetgum tests, spring treatments were most effective in both top-kill and inhibition of sprouts. Treatments in sommer were better than those in fall and winter (Fig. 1 ). Similarly, on trees not completely dead, spronts were fewest and smallest after spring treatment While the differences between seasons were not statisti­ cally significant, they are consistent with those reported by other work­ ers (5, ff). Without chemicals about 75 per­ cent of the post oaks sprouted after girdling as compared with about 40 percent of the sweetgums. The dif­ ference was not statistically sig­ nificant, and is inconsistent with considerable experience which rates sweetgnm the more difficult species to controL The tests were confined to a rath- er narrow range of diameters—3.6 to 7.5 inches d.b.h.—because these size classes have generally sprouted most vigorously after girdling. Within this range, cLbJh. of tree was significant only in the percent of sweetgums top-killed in the sea­ sonal test. Here smaller trees were top-killed more effectively than large ones. Differences were largest where no chemical was applied. Conclusions A x girdling was significantly bet­ ter than power girdling only in topkill without chemical treatments. With chemicals there was no dif­ ference between ax and power gir­ dling. Top-kill differences reflect promptness rather than eventual completeness of kill, since remain­ ing tops are moribund. A ll treatments involving 2,4,5-T resulted in significantly higher top- kill than girdling alone. Concen­ trated and dilute 2,4,5-T killed crowns equally well; the concen­ trate, however, proved to be a poor sprout inhibitor, while the dilute solution was very effective. There was no significant differ­ ence between the response of sweetgum and post oak. Within the limited diameter range of the tests, size of tree did not affect results significantly. Dilute 2,4,5-T was most effective in reducing sprouting when applied in early spring, least effective in the dormant season. It is evident that the saving in material costs and transportation of dilutent envisaged in the design of this study cannot be fully achieved by application of undi­ luted 2,4,5-T. Where promptness of top kill is a primary considera­ tion, application of full-strength 2,4.5-T in cambium girdles may prove usefuL Where both prompt kill and inhibition of sprouts are required, the chemical should be ap­ plied in dilute form. Further tests are needed to deter­ mine whether some intermediate concentration of 2,4,5-T can be ef­ fective without being too bulky for use with a power girdler. Literature Cited 1. C ampbell, B. S., and I t o A. P ervi . 1950. Chemical control of undesirable southern hardwoods. Jour. Range Mgt. 3:118-124. 2. D avis, J . B. 1958. Diluted 2.4,5-T more lethal than undiluted in east Texas. Jour. Forestry 56:516. 3. Grano, Charles X 1958. Besponse of southern red oak to seasonal appli­ cations of 2,4,5-T. Jonr. Forestry 56: 140-141. __ 4. M n m r , A. L 1956.. W hat gives with girdling t Southern Lumberman 193(2417) :214-215. 5. P eeyt, P eed A. 1956. Methods for controlling hardwoods. Fornata and People 6(3) :22-25, 34-35. 6. S hipman , B. D. 1958. Effect of sea­ son of treatment on girdling and chemical control of oak and sweetgnm. Jour. Forestry 56:33-35. 3S 5268 3 $ T -C N ' THE DOW C H EM IC A L COMPANY ^ y VT^c’r BIOCHEMICAL RESEARCH LABORATORY J O U R N A L A B ST R AC T K ~ - ( ( 0-t>) SUBJECT») OR COMPOUND«). 4 AUTHOR»)! f./O Z G J - (> h a J l .L u . I j P s5 f t . A . f & . C-n 03 h ')' CO cn -7 0 U ___ > ABSTRACT 6 c m ^ C c * » ■z j u o ^ M - ¿ t'V M * * * , u ~ * . , ¿ - - V / q --- -------- 1 / i f + .X -9 * u ^ e^ _ u * . . ^ Uk i z ¿ v t y . t 7- /3 ' c -r '~ f /-/- i A '* i^»-r r ~ > * - / * * - ? : / / . ? ? ^ ' y ' ^ « r £ S ROUTE TOt LABORATORY! « 4 e0 « W -rffeO. « WfjTC « .i« a r ~ fc e .i.- 0009999 8T1 5269 ^■ m . T y fc s fC . DATE: "■ < e - « j i FILE : Piiirto ii u. s . a. t-g monoiodoacetate glycolysis The result was the stimulation a of without diametrically ph y tohormoyies which which it are an­ [3]. Fundamental principles Optimal doses of auxins like indoleacetic acid and 2,4-dichloro phenoxyacetic acid can increase oxygen consumption of vegetal tissue from 20 to 60% [4]. hydrate depletion. This respiration increase accompanies a carbon Strong concentrations inhibit g r o w t h and r e s p i r ­ ation in the same way as fermentative glycolysis [5]; but r e s p i r a ­ tion can "still be.kept elevated at those doses which inhibit growth. Auxin activity is thwarted by reagents of -SH groups such as io d o ­ acetic acid. Thus auxins act through intermediary sulfhydryl e n ­ zyme groups, most particularly those dehydrogena cycle. -S of the Krebs Interaction with the -SH group of coenzyr.e A should be for- seen as possible. Auxin action resembles coenzyme action. It p a r ­ ticipates in m o s t phosphorylation systems and contributes to the aerobic transport of phosphates [6]. to fraction I vegetable proteins. In plants, auxins are bound This fraction has a phosphatase activity and can, along w i t h animal phosphatases,rcatalyze phosphorylations [7]. In stimulating concentrations, trans­ auxins p r o ­ mote water absorption by vegetable cells, absorption whi c h depends on an oxidative metabolism and very particularly -^n oxidative phosphorylations [8]. Moreover, these substances provoke in tis­ sues a considerably diminished potassium rate whi c h is elevated in „ 5272 0007451 È Page 2 f malign tumors. But their m o s t remarkable property is to influence the decisive work of the nucleic acid content of cells. DMA and RNA rates increase or abate as a function of auxin c o n c e n t r a t i o n [9}. r Finally, auxins are antagonistic to such cell division factors as adenine and kinetin [1]. Pa rticularly interesting is the action of phytohormones on the tissues of cultured vegetable tumors ("crown galls") . concentrations totally lack a stimulating action other hand, [11]. Weak On the the grov/th of tumorous sunflower and periwinkle t i s ­ sues lessens in the presence of we a k concentrations of i n doleacetic acid, indolebutyric acid and 1-naphthaleneacetic acid. More e l e v a -4 ted concentrations (1 x 10 ) totally inhibit this growth. Such concentrations also stimulate normal sunflower tissue [12]. At -4 1 x 10 , indoleacetic acid stops the wet weight increase of t u ­ morous tissues and tissues "accustomed" to scorrcnera [13]. The 2,4-D inhibits tumorous sunflower tissue at concentrations of 1 x 10 * 13 J , and tumorous *5 tobacco tissue from 1 x 10 only. A u x i n action in plants increases as they gr o w more rapidly; young tissues are more susceptible than old ones. same plant, In one and the the less mature and less differentiated cells are the mo s t responsive. . Struckmeyer and his collaborators [14] have histologically o b ­ served transformations of tumorous sunflower tissues under the i n ­ fluence of auxins. Weak concentrations have not, on the whole, modified wet weight but have diminished m e r i s t e m cells per surface u-nit and have increased scalariforn vessels. tions, A t strong c o n c e n t r a ­ the number of cells of all types greatly d i m i n i s h e d and the w e t weig h t fell impressively. Proof In ’spite of these facts, auxins have been neglected in the search for cytostatic substances, sons- no doubt for the following r e a ­ The mere name "growth substances", implying a g r o w t h s t i m u ­ lant, may be one of the causes of their omission. nations are deceptive, centration, But such d e s i g ­ since auxins can, according "to their c o n ­ also stimulate as well as inhibit growth of vegetable 527:b o o ? 4 5 2 Page 3 » tissue, and since weak concentrations, again, inhibit tumorous tissue. Besides, auxins have attracted to themselves the reputation of being carcinogenic. Previously the formation of a callus at benign hyperplasia. ment: Gautheret [15] advanced a m o s t serious a r g u ­ From a carrot tissue whi c h he had cultivated for m a n y years liUl/V I the surface level appeared suspect, even though it was really a in a cultured m e d i u m increased with 0.1 mg/£ indoleacetic acid, Gautheret isolated a cellular strain able to continue to gr o w and proliferate without any additional indoleacetic acid. This new tissue, which he called "habituated tissue", was friable and trans lucid, consequently morphologically modified. Grafted onto a 4 healthy plant, it became a true tumor of "habituated" tissue, [16j . This transformation apparently cancerous tissue, was g enerally attributed to a u x i n ’s influence. this type of perception. It is acceptable today to doubt In effect, Sanford and his collaborators, starting from a single mesenchymal cell, successfully isolated at the end of twenty months a cellular strain ("clone") which, in 97% of the cases, instead of forming sarcomas, inoculated the cells of healthy mice [17] . Other authors likewise have obser v e d i n v i t r o such a transformation of normal mammal cells into cancerous cells [18 The appearance of malign i t y lasted from four months up to four years. Goldblatt and C a meron attributed the malign appearance to an a n a e r o biase. In none of the other cases was it possible to determine the cause of a given carcinogenic agent. fallowing conclusion: In fact, Sanford came to the all cell strains, cultivated in a hete r o ­ logous medium for a sufficiently long time, finally acquired the p roperty of giving birth to either sarcomas or carcinomas when these cells have inoculated animals of the species fr o m which the culture originated. In other w o r d s ,*every tissue, culture is event- • ually carcinogenic. Two other facts have provoked the thought that phytohormones can be carcinogenic: a) table tumors, produces the B a c t e r i u m tunefacisns, agent of v e g e ­ indoleacetic acid in an a rtificial medium; b) vegetable tumors contain more auxin than corresponding healthy Page 4 tissue. However, indoleacetic acid production by 3. reaches at most 125 nicrograms per liter iumefacisns [19], when 10-30 million micrograms is necessary to artificially provoke a gall [20]. Other bacteria also produce auxins in an artificial medium; but they never instigate the formation of tumors [21]. The high auxin rate verified in vegetable tumors is due to the fact that because of their incomplete oxidations, they consist of more free auxin and less bound auxin in protein fractions. The auxin- protein connection depends in effect on normal oxidations and 1 the presence of adenosinetriphesphate [22]. A n o t h e r reason for eliminating auxins from the list of actual cytostatics is the fact that these are substances whose action is strictly limited to vegetable metabolism. However, past and p r e s ­ ent publications report on work proving their influence in animal metabolism. We recall the discovery of the regular presence of auxin a and indoleacetic acid in human urine [23] whi c h was a t t r i ­ buted to the action of intestinal bacteria. In 1949, Raoul and M a rnay verified a growth stimulation in young rats under the e f ­ fect of w e a k doses of indoleacetic acid [24] . Zambotti and De B e r n ­ ard thr ew into relief indoleacetic acid's influence on r e s p i r a t i o n and glycolysis in'animal cells [25] . Finally the works of Louis have shown an impressive parallel between indoleacetic acid's effect on the cultured tissues of vegetables as well as animals. In a n a e r ­ obic conditions, weak concentrations have been stimulants and strong concentrations have been inhibitors to cultured cardiac cells of chicken embryos. In aerobiosis, only inhibition by v e r y strong doses was observed. division; mitoses Indoleacetic acid distinctly works on nuclear increasing concentrations severely diminish the numb e r of [26] . Thus, I have led up to the search for whethet auxins, whose cytostatic effect on "crown galls" has previously been pointed out, jT can as well inhibit animal and human tumors. 0007454 > 5275 Page 5 Toxicity G i ven the massive use of auxins in agriculture, has been tested several times. their toxicity Six grams of 2,4-dichlorophenoxy- acetic acid was administered to cows for three and a half months without provoking the least symptoms of intoxication. A research worker has taken 0.5 grams of the same substance for twenty-cne days w i t h o u t disturbance. In spite of this, icity, I veri f i e d one m o r e instance of 2,4-D t o x ­ this time in the form of ethyl ester, among rabbits. At •first, a dose of 0.01 g/kg of weight was injected intramuscularly. A f t e r five days, the dose was increased to 0.02 and, after another five days, to 0.05 g/kg. tieth day. The experiment was stopped at the t w e n ­ The appetite and vivacity of the animals were not at all impaired as the doses were raised. As a continuation, I par- enterally administered auxin to animals and humans in average daily doses of 3 to 4 mg/kg for very long periods, without ever observing toxic phenomena or undesirable reactions. A patient re• ceived, m for one hundred forty days, an ethyl ester of 2,4-D, then an isopropyl ester of 2 , 4 ,5-trichloropher.oxyacetic acid in daily intramuscular doses of 0.10 to 0.30 grams with o u t presenting symptoms of intoxication. olive oil solution. These products were employed in an Intragluteal injections caused neither burns m nor aches and were easily reabsorbed. During the course of a year, the hundreds of injections never provoked an abscess. *•■■**. ■ Concerning administration p e r o s , daily quantities of 0.30 g and more soon released a laxative effect. it should be noted that (divided into doses of 0.10 g) Other subjects, particularly those given 2,4-D and 2,4,5-T esters, experienced a disagreeable gout. The fact ought to be emphasized''that hematopoietic organs and the blood formula v/as not influenced. i The rat e s of chole s t e r - ol, urea, and glucose in blood were never augmented. hypoglycemia was occasionally verified. ho time raised during the treatment. Only a slight A r t e r i a l pressure was at We are dealing with a group 0007455 5276 Page 6 of ve r y weakly toxic s u b s t a n c e s , not having, in opposition to the antimetaboliies and nitrogen mustard derivatives, any effect on hematopoiesis. After not having ever exceeded daily doses of 0.45 g from October 1956 until October 1957, I decided to use daily quantities of 1.25 to 2.50 g (once at 1.25 to 1.50 or twice at 1.0 to 1.25 g intramuscularly). Even at these very elevated doses, the injected substance was painless and easily reabsorbed; on the other hand, certain disturbances soon appeared. They consisted of a lassitude, vparticularly in the legs, a heightened thirst, vertigo, two cases, a slight confusion. in The last two symptoms did not a p ­ pear at the mo s t elevated doses. certain inebriety. and, The patients exhibited then a Twice I noted diarrhea, and once, vomiting, A few days after the interruption of treatment, disappeared completely. the toxic symptoms As a notable secondary effect during treatment wi t h these elevated doses, arterial pressure mark e d l y diminished. ■ Drill has pointed o ut among mammals the following toxic s y m p ­ toms: weakness, diarrhea, weight loss, and, in certain species, a curious state of a myot o n i c response to quinine [27]. Preliminary analyses The initial analyses were attempted with incoleacetic acid and the sodium salt of 2,4-dichlorophenoxyacetic acid. The first was m i x e d with equimolecular quantities of piperazine, rendered thus hydrosoluble, and injected intravenously in a 1 to 2% solution at a r a t e of 0.02 to 0.10 g/day. The second, h y drosoluble naturally, was administered in the same fashion in a dose of 0.10 g/day. cases (a lymphogranulomatosis, tic c ar cinoma mastitoides, stomach cancer, i.e., Hodgkin's disease, a metasta­ a lymphosarcoma, a m u l t i p l e m et a s t a t i c and an adenbcarcinoma of the prostate w i t h post- o peratory métastasés) were thus treated for twenty-five days w i t h ­ out either amelioration or aggravation. Neither of the two sub­ stances provoked the appearance of toxic symptoms. were painless, Five non-hardening, The injections and did not release, s e c o n d a r y ^ i ^ c t i o n s 000745« Page 7 These analyses seem to confirm the inactivity of auxins in humans in contradiction to the qualified observations on the c u l ­ tured tissues of animal origin. human blood pH One can ask if, at a very elevated' (pH of human blood: 7.3 to 7.4; pH of vegetable t i s ­ sues:: 5. 5 to 6.0), auxins have not been dissociated to the poi n t of losing their activity. Effectively, Audus has been able to verify that a weak dissociation of 2,4-D is linked to an elevated toxicity for plants. were dissociated. Already at a pH of 3.28, 50% of the m o l e c u l e s Audus admits that 2,4-D works uniquely as a non- vdissociative molecule [231. Among the derivatives of indole a c e t i c acid and phenoxyacetic acid, the alkaline salts and the amid e s are ionized in the same style as the acids; only the ester and nitr i l e molecules and the corresponding aldehydes continue to be nondissociative. Analysis is thus followed up for these latter categories, principally with the esters. Analyses with non-icr.icable auxins Derivatives e m p l o y e d : Fr o m October 15, 1956 to October 11, 1957, auxins w e r e a d m i n ­ istered in daily doses of less than 0.45 g . .1 successively e m ­ ployed the ethyl ester of 2,4-dicnlorophenoxyacetic acid, isopropyl ester of 2,4,5-trichlorophenoxyacetic acid. exists on the m a r k e t in the form of a brown liquid, proportion of vegetable oil. (50%), and the The first soluble in any Even at the strongest co n c en t r a t i o n s the solutions stay clear and resistant to sterilization. The second appears in the form of a light brown crystal composed of a compact mass- that ought to be melted at 8 0° to 90° to be able to d i ssolve in olive oil. Mixed in cold oil, iately recrystallizes; but after reheating solution stays stable, at .least up to 15%. By October 1957, the substance i m m e d ­ (sterilization), the the impression became clear that a d a i l y dose of 0.10 to 0.20 g of these esters could not bring abo u t in humans conditions which corresponded to a concentration of 1 x ±0 .to 1 x 10 -7 because such concentrations were the m i n i m u m rate necessary to obtain total inhibition in a vege t a b l e tumor culture. i 5278 0007407 Page 8 One m u s t in reality administer doses near 1.0 g at least and, tained above 1 mg % for the w h o l e day, p e r dicrr. q uantities of 1.50 the administration of such propyl esters of 2,4,5-T given its limited solubility. I am o b ­ liged thus to return to 2,4-D ethyl ester; but I have r u n up 4 against the toxic troubles I indicated before. I am f o r c e d then to find more active derivatives that allow a d m i n i s t r a t i o n ’of ^smaller quantities. The ethyl and methyl esters of 2,4,5-tri- chlorophenoxyacetic acid fulfill these conditions. . 6 0 6 Î 1 Ê C quantities is not possible wit h 25 to 50% solutions or w i t h iso­ M M However, I to 2.50 appear appropriate. " since it is important that the rate in the interior level be m a i n ­ Phytobiological tests fixed at first an acti v i t y five times greater than that of 2.4- D ethyl ester. Thes e w h i t e and compact needle crystals do not have a limited solubility in oli v e oil. Instead of the projected 30% solution, a 20% solution wa s prepared wi t h the addition of 5% acetonitrile. The substance recrystallized however and it had to be reheated in a boiler to redissolve it before each injection. Above 30° the solution was stable. phytobiological or otherwise, 2.4.5- T methyl ester. None of the e x p erimental data, has been published c oncerning the One knox*/s nevertheless that the esters of phytohormones inhibited germi n a t i o n and budding m o r e as the r a d i ­ cal alcohols of these esters allowed the number of carbon atoms more ^ riance. The white 2,4,5-T crystals of m e t h y l ester are again less soluble than those of ethyl ester. The r e c r y s t a l l i z a ­ tion of a prepared 10% solution required reheating in a boil e r b e ­ fore injection. The fear that instigated the u se of the m e t h y l ester was now confirmed. The m e t h y l 2,4 , 5-trichlorophenoxyacetat e could be administered parentarally in doses of 0.85 g per day for six weeks without provoking the least apparition of trouble. r The effects of the two primary derivatives e m p l o y e d (.ethyl 2.4- dichloropheno^yacetate and isopropyl 2,4 , 5-trichlorophenoxyacetat e tate) we re g r a c s o rr.odo the same and the second did not exceed by a bit .ne first in efficiency. The ethyl and meth y l esters of 2.4.5- trichloronnonoxyacctic acid proved to have a cytostatic activity superior to that of the isopropyl ester. This precedence 5279 0007458 Page 9 I inspired confidence in methyl ester whose tolerance was perfect. Successfully ameliorating its solubility will allow highly e l e ­ vated doses to be given. Clinical e x p e r i e n c e : Of the 41 cases treated, 5 were animals. The auxins used had a particularly m a r k e d effect on b e n i g n turners. In four cases of fibroadenoma of the prostrate, troubles (retention, dysuria, vtant urination) the pains, and stuttering and precipi- disappeared during the first week. A t the end of ten to fiftèen days, hypertrophy had retrcceeded in such a m a n n e r that the treatment could stop on the fifteenth day. Efficacy was also r emarkable on a recently appeared diffuse adenoma of the right lobe of the thyroid. After fifteen injections of 0.15 g of 2,-4,5-T isopropyl ester, a goiter, as large as a prune, disappeared. A fibremyema of the uterus retrcceeded, ment, during twenty days cf t r e a t ­ from the size cf a grapefruit to that of a detectable pain curve. On- cancerous tumors of reduced m a l i g n i t y and with o u t d e t e c t ­ able metastases, efficacy was equally remarkable; amelioration and, -in three cases, recovery there was always (one facial fibroepi- thelioma in a donkey and two epitheliomas of the type u l c u s r o d e n s in a dog and a c a t ) . 4 In cases of average malignity, there was a clinical reco v e r y (tumor at the point of cholecystic parting in a 55 yea r old w o m a n wi'th enormous metas t a s e s of the mesen t e r i c ganglia), ameliorations, m o m e n t a r y halts, or a slackening of the process occurred. cases of great mali g n i t y coast type) The (for example pulmonary cancer of the P a n ­ or preagonic m a l i g n i t y reacted hardly or not at all. It is noted that at doses less than 0.45 g, the esters of 2,4-D and 2,4,5-T ^ e r e perfectly well supported by infants. seven ye a r old boy, A afflicted w i t h a m a l i g n lymphogranulomatosis, r eceived a daily dose of 0.10 g for three months w i t h o u t any u n d e ­ sirable secondary reaction. 5280 0007459 Page 10 Among the treated cases were mammary ad e n o c a r c i n o m a s , b r o n ­ chogenic carcinomas, rectal cancers, epitheliomas, lymphosarcoma wi t h multiple tumors, Radiotherapy a little a lymphoma, a etc., but no leukemia. (less than three weeks) prior to auxin treatment increased tumor sensitivity in these lymphosarcomas. A remarkable effect was observed in a m a n affected with a b r o n c h o ­ genic carcinoma who, a few days before treatment with 2,4-D ethyl ester, h a d received 1500 r in six sessions. old woman, A fifty-eight year suffering from an inoperable cancer of the uterus, re­ acted in such an extremely favorable manner at that time to 2,4-D ester treatment that she then received a total dose of 11,480 r.' After 48 injections starting on December 24, 1956, she remains r• clinically healed. Radiotherapy and treatment with esters of auxins, applied simultaneously, seem to have a reciprocal potential. A proof was furnished b y a 56 year old woman afflicted with a lymphosarcoma with multiple tumors. A single treatment of 2,4-D ethyl ester gave an amelioration; but the patient had developed a resistance and did not react to a second series. was begun again, On April 4, 1957, treatment this time with 2,4,5-T isopropyl ester at a rate of 0.10 g/day intramuscularly, simultaneous with X-ray radiation. Although the patient received in all, as of May 12, 1957, 2100 r (fifteen sessions of 125-150 r under 175 kw.with 1 m m Cu and 1 m m Al, distributed across four areas, axillary), r^ult, only two cervical and two the gangliary tumors nearly totally retroceded. This for a second radiotherapy and considering the given dosage, was unforseen. In another case, also treated simultaneously wit h X-rays and a 2,4,3-T isopropyl ester, a total dose of 2250 r spread over twenty-two days sufficed to completely dissipate Hodgkin's tumors. This also was a question of radiotherapy repe a t e d for the second time. In a third case, where the efficacy of esters of auxins seemed to h6ve been exhausted, a simultaneous radiotherapy resulted in new and palpable progress. In an osteosarcoma of the thigh, on the other hand, simultaneous radi.._ion was a disastrous influence and had to be stopped. > 52810007460. Page 11 The efficacy of esters of auxins was at a m a x i m u m for the first two weeks; then began to decline; for one of two weeks. gress at all. After a month, but stayed still notable there was hardly any p r o ­ Wi t h 2,4,5-T isopropyl ester, perhaps a little faster. efficacy diminished It seems thus to develop a certain re ­ sistance. simultaneous administration of riboflavin p e r c s and intravenously. Note that the auxins employed w e r e contaminated by polychloro-. phenol. These latter considerably augmented the action of auxin- oxidases by whi c h furthermore they induced or amplified formation [29 It is possible that these polychlorophenols are in part r e s p o n s i b l e for the resistance provoked above. As w i t h 2 , 4-dinitrophenol, they disconnect respiration, prop e r l y called oxidative phosphorylations, and prevent the formation of ATP. However, the commercial esters of auxins of w h i c h I availed myself continually b r i n g to mind i s o ­ mers such as 2,6-D and 2, 4 , 6-T. of active auxins. Also, These isomers inhibit the action in the clinical analyses w h i c h have b e e n attempted in a Parisian hospital, we exerted ourselves to util i z e substances carefully purified by recrvstallization in the appro* priate solvents. .We further hope to be able to test, on an index of raised activity, other auxins which I was not able to prepare in sufficient quantities. Discussion In 1953, Silberberger and Skoog [9] proved that auxins like indoleacetic acid wo r k in a direct fashion on the amou n t of nucleic acids in vegetable tissues and that growth depends to a large e x ­ tent on this relation. The amount of ribonucleic acid and d e o x y ­ ribonucleic acid in a vegetable tissue cultivated i n v i t r o begins to increase at th^ moment when increasing q uantities of indo l e ­ acetic acid are added to the culture medium. For d e o x yribonucleic acid, for r ibonucleic acid, this increase is optimal at 0.014 mg/1; it is only optimal at 3.3 mg/î.. Exceeding the optimal doses of indoleacetic acid diminishes the amount of nucleic acids. At > 5282 0007461 t n # h 6 •/ The efficacy of esters of auxins regularly increased through Page 12 90 mg/1, it ought to be lower than that of the controls and finish by r e a c h i n g — 50% and less towards 270 mg/&. Increase and decrease of this amount parallels the fluctuations in wet weight. call here S t r u c k m e y e r 's observations of vegetable tumors. We r e ­ [14] on the tissue cultures He remarked on a considerable drop in the w e t w e i g h t under the influence of strong concentrations of auxins. We also recall here that 2,4-D ethyl ester is more active than indoleacetic acid and that tumorous tissue is only sensitive to the inhibitory effect of phytohermones. We can thus suppose that at ■elevated concentrations of certain auxins, the tumor's amount of ribo- and especially deoxyribonucleic acids can be reduced. This is particularly important if one bears in mi n d the decisive role that ribo- and especially deoxyribonucleic acids play in the d i v i ­ sion of nuclei and cells. Fro m 1953 to 1956, Miller [30], sta r t ­ ing wi t h a preparation of deoxyribonucleic acid from herring sperm, successfully isolated a highly cytokinetic substance, whose p r e s ­ ence was impartially verified in analogous preparations from the thymus of a calf and in yea s t extract. This factor, w h i c h he called kinetin, was capable of r e l e a s i n g cellular division in a concentration of one yg/Jl. 6-furfurylaminopurine. It turned out to be a puric derivative, Fresh preparations of deoxyribonucleic acid did not exhibit kinetin action; this only appeared after a bandoning the preparation for months or after a sojourn of sixty minutes in an autoclave at 125°. The kinetin seems to be a p r o ­ duct of deoxyribonucleic acid disintegration and it is correct to svippose that its presence in tissues is strictly d ependent on the amount of deoxyribonucleic acid. It thus seems justified to p r o ­ pose the hypothesis according to whi c h the cytostatic action of an auxin is explained by the rarefaction of cell division factors such as kinetin. From the work of Skjpog and his school follows that, of t$ie two elements of v egetable growth, and cellular division, concentration; [31] , it elong a t i o n the first depends o n having all of the auxin the second, on the factors such as kinetin or an integral part of this 'last, adenine. A modifi c a t i o n between the proportions of adenine and auxin favoring the first expresses r** a ( 5283 o o o v is; I Page 13 itself by an increased DMA content; a modification favoring auxin finds its expression in an elevation of the RN A content. This leads us to propose the following equation: Adenine (kinetin)/auxin = DMA/RUA = Skoog and Tsui have shown that elevated adenine concentrations I and w e a k auxin concentrations favor budding and that on the c o n ­ ** f division/elongation. centrations [32]. This antagonism between the factors of cellular division such as adenine dines like guanine, ' hand, (which can be potentialized b y pyrimi- • thymine, and cytosine) and kinetin on the one and the factors of elongation such as indoleacetic acid on the othe r hand, is of a very peculiar nature. In fact, kinetin is only active in the presence of traces of auxin [33]; inversely, it seems that auxins need the presence of adenine to have an effect on cellular elongation [34]. Still, a great preponderance of the one can stifle the characteristic effect of the other [35]. It is perhaps daring to transpose into the domain of animal 1 biology facts discovered in that of phytobiology; however, for the mom e n t these latter alone can help us understand the effect of auxins on tumors in humans and animals. Summary In w e a k concentrations, auxins accelerate the catab o l i s m of carbon hydrates, activate oxidative p h o s p h y l o r a t i o n s , increase o x ygen consumption in tissues and stimulate growth. concentrations, they complete the depletion of carbon hydrates, obstruct anaerobic glucolysis, tissues, In elevated and inhibit growth. reduce the rate of p o t a s s i u m in Auxins ^regulate the r a t e o f nucleic acids and are antagonistic to factors of cellular d i v i s i o n such as k i netin or -6-furfurylaminopurine. growth is remarkably selective. Their inhibitory action on It is most evident for certain vegetable species than fwx others and, in all cases, stronger as the tissue in which it operates is younger and less differentiated. 1 52840007463 n r ^ trary budding was inhibited by elevated auxin and we a k adenine c o n ­ Page 14 The utilization of auxins as synthetic herbicides is bas e d on these properties. Tissues of vegetable tumors are more sensitive to auxin's inhibitory action than corresponding normal tissues. Auxins operate, bolism. although to a lesser degree, on animal m e t a ­ The author has undertaken to utilize certain auxins for the selective inhibition of b e n i g n and m a l i g n a n t tumors in humans and animals. Auxins in ionizable form prove to b e inactive. the contrary, non-dissociable derivatives manifest considerable cytostatic action. dichlorophsnoxyacetic acid, phenoxyacetic.acid, (esters of alcohols) The ethyl ester of 2,4- the isopropyl ester of 2,4,5-trichloro and especially the met h y l ester of the same acid are particularly active. The a nti-tumor effect of these a u x ­ ins is reinforced by riboflavin. slightly preceding On Radiotherapy concomitant or (less than three weeks) treatment w i t h auxins considerably reinforces the cytostatic action of this last, even w h e n the dose of rays is subliminary [sic]. The cited esters of auxins have a very slight toxicity and never depress hematopoeisis T h e i r hyperglycemic effect was confirmed. They we r e p r e f e r ­ ably administered in an oily solution by intramuscular injections of 0.50 to 1.0 g/day. Rests of four days can be i n tercalated b e ­ tween successive series of fifteen days. The esters of auxins utilized m u s t be purified of any c o n t a m ­ ination by the corresponding polychlorophenols a n d * neighboring isomers, whether inactive or antagonistic. Acknowledgments A m o n g the treated cases, m y thanks to Dr. E. C. Preston Animals, Tangiers) there w e r e five animals. I direct (The People's D ispensary for Sick for having permitted the treatment of them. I equally thank h i m for the efficient* help and enligh t e n e d counsel that he so often squandered on me. However, I have to express my gratitude to Messieurs the Pr. H. V e l d s t r a and H. Linser at the same time as to the Rhone Poulenc Laboratories, and Phili s-Roxan for the substances which they freely consented to put at my d i s ­ posal. 5 2 8 5 0 0 0 ? 4 6 4 _ Page 15 [1] O. Warburg, N a t u r w i s s c n s c h . , £1, 483 (1954); £2, 401 (1955). [2] E. Lundsgaard, Am. Rev. B i o c h e m . , 1_, 377 a n d W. D. Bonner, Am. J. Botany, 36_, 214 [3] J. Berger, P. Smith and G. S. Avery, Jr., Am. J. Botany, 601 (1946). [4] G. S. Avery, Jr., Plant Growth Substances, p. 105, F. Skoog, Univ. Wiscons. Press, M a d i s o n (1951). -[5] F. G. Smith, Plant Physiol., (1938); K. V. Thimann (1949) . 33, 23, -70 (1948). C J. Bonner, Am. [7] B. Axelrod, J. Biol. C h e m . , 1 7 2 , 1 ' [81 [9] J. Botany, 36, 323, 429 «It o c i V [6] (1949). (1948). D. P. Hackett and K. V. Thimann, Proc. Nat. Acad. Sci., 3 8 , 770 (1952); J. Bonner, R. S. Bandurski and A. Millerd, Physiol. Plantarum, 6, 511 (1953). J. S i l b e r b e r g e r , Jr. and F. Skoog, Science, 1 1 3 , 443 (1953). [10] F. Skoog and C. Tsui, Plant Growth Substances, p. 263, Univ. Wiscons. Press, Madison (1951). [11] R. S. De Ropp, Am. J. Botany, 3£, 248 C. R. Acad. Sci., 22£, 1728 (1950). [12] R. S.^ De Ropp, Am. J. Botany, [13] R. J. Gautheret, 3£, 53 C. R. Soc. Biol., (1947); R. J. Gautheret, (1947) . i£2, 774 (1948). [14] B. E. Struckmeyer, A. C. Hildebrandt and A. J*. Riker, Am. J. Botany, 36, 491 (1949). [15] R. J. Gautheret, Bull. Soc. Chim. Biol., 2£, 13 (1942). [115] G. Camus and R. J. Gautheret, C. R. Acad. Sci., 2 2 6 , 744 [17] K. K. Sanford-, G. D. Likely and W. R. Earle, J. Nat. I n s t . , 15, (2), 215 (1954). (1948). Cancer [18] H. Gold b l a t t and G. Cameron, J. Exper. M e d . , £7, 525 (1953); W. R. Earle and A. Nettleship, J*. Nat. Cancer Inst., £, 213 (1943); G. O. Gey, M. K. Gey, W. M. Firor and W. 0. Self, A c t a Unio. Intern, contra Cancrum, 6, 706 (1945); K. K. Sanford, W. R. Earle, É. Shelton, E. L. Schilling, E. M. Duchesne, G. D. L i k e l y and M. M. Becker, J. Nat. Cancer Inst., LI, 351 (1550). [-19] S. B. Locke, A. J. Riker and B. M. Duggar, Acir. Res., 535 (1939). " 59, 519, .5286 0007465 . \ Page 16 II. Berch and B. M. Duggar, J. Agr. R e s . , 6 3 , [20] A. J. Riker, 395 (1941). [ 21] S. B. Locke, A. J. Riker and B. II. Duggar, J. Agr. 519, 535 (1939). [ 22] S. M. Siegel and A. W. Galston, _39, 1111 (1953). Proc. Nat. Acad. Res., £9 t Sci. Wash. [23] F. K o e g l , A. J. Haagen-Smit and H. Erxleben , Z. Physiol. Chem., 228, 90 (1934). [24^ Y. Raoul and Ch. Marnay, Bull. Soc. Chim. Biol., 31, 289 (1949) [25] B. De Bernard, Boll. Soc. Ital. Biol. S p e r . , ££, 81 (1952); V. Zambo'tti and B. De Bernard, Boll. Soc. Ital. Biol. Sper.’, 28, 928 (1952). [26] R. Louis, Zeitschr. f. Zellfschg., 41, 385 [27] V. A. Drill, Pharmakology in Medicine, Co., New-York, Toronto, London (1954). L. J. A u d u s , N e w Phytologist, £8, 97 (1955). 60/7, McGraw-Hill Boo k (1949). [29] P. L. Goldacre, A. W. Galston and R. L. Weintraud, Arch. chem. Biophys., £3, 358 (1953). Bio- [30] C. C. C. F. O. Miller, Proc. Soc. Exp. Biol, and Med., £3, 561 (1953); 0. Miller and F. Skoog, Âm. J. Botany, £0, 768 (1953) ; or Miller,. F. Skoog, F. S. Okumura, M. H. Von Saltza and M. Strong, J. Am. Chem. Soc., 7£, 1375 (1956). [31] F. Skoog, Brookhaven Symposia in Biology, £, 400 [32] F. Skoog and C. Tsui, Plant Growth Substances, Wiscons. Press, Madison (1951). (1954) . p. 263, Univ. [33] C. O. Miller, F. Skoog, M. H. Vo n Saltza and F. M. Strong, J. Am. Chem. Soc., [34] C. O. Miller, 77, 1392 Proc. (1955). Soc. Exp. Biol, and Med., £3, [35] F. Skoog and C. 0. Miller, Symposia Soc. Exptl. 118, Cambridge Univ. Press (1957*). 561 Biol., (1953). 11, ( 000746*. 5287 5288 H-o ■ 5291 «EF SERVICE J o u r n a l o f th e W e e d S o c ie ty o f A m e r ic a 5292 BR IEF PAPERS Iso-octyl Ester of 2 ,4 ,5 -T in Hardwood Control Conctntraiion (lb ahg) C a r t e r B . G ibb s 1 40. . 20. . an d D u k e (2) in 1955 reported su ccessfu l u se o f a 36 lb ah g so lu tio n o f th e p ro p y len e glycol b u ty l eth er ester o f 2 ,4 ,5 - T a p p lie d by'‘m ean s o f a tree in jecto r.- T h is to o l is essen tially a m o d i­ fica tio n o f th e C orn ell to o l describ ed by C o p e and S p aeth (3) in 1931, a n d su b se q u en tly tested w ith a m m o n iu m su lfa m a te (I , 5) and 2 ,4 ,5 - T (4, 6, 7). S in ce th e iso-octyl ester o f 2 ,4 ,5 -T is g en er a lly a v a ila b le a t lo w er cost th an th e p rev io u sly tested p rop y len e g ly c o l b u ty l eth er ester, a stu d y to assess its effectiveness was in sta lled in M arch 1957 by the N a co g d o ch es R esearch C en te r in co o p era tio n w ith th e Sam H o u s­ to n N a tio n a l Forest. E ach o f fou r co n cen tra tio n s o f th e ch em ica l, 40, 20, 13.3, an d 8 lb a h g in d iese l o il, w as a p p lie d to 30 sw eetgu m s (L iq u id a m b a r styraciflua L .) an d 30 oaks ( Q uereus stellnta W ang en h ., Q. m a rila n d ica M u en ch h ., and Q . falcata M ich x .). T r ees w ere o f tw o d ia m e te r classes, 0 .6 -3 .5 inches a n d 3 .6 -6 .5 in ch es. A to ta l o f 480 trees w er e th u s in jected — all in circum basal w ou n d s, o n e p er in c h o f estim a ted tree d ia m eter. S in ce an a im o f th e stu d y w as to e x a m in e th e effectiveness o f variou s con cen tra tio n s u n d er field co n d itio n s, n o sp ecial m easures w ere tak en to assure ex a c tly u n iform a p p lica tio n s o f the silv icid e. E ach treated tree w as in sp ected for crow n c o n d itio n an d sp ro u tin g in M ay a n d O c to b er 1957 an d in N o v em b er 1958. A t th e first in sp ectio n , th ree m on th s after treatm en t, crow n dam ­ a g e w as v isib le o n ly on trees treated w ith th e h ig h e r con cen tration s. B y th e en d o f th e first g r o w in g season th e m o st effective treatm ent, 4 0 lb ahg, h a d to p -k illed 100 p ercen t o f th e sm a ll trees and 93 per­ c e n t o f th e large trees. T o p -k ill was slow er o n sw eetg u m th a n o n oaks. N o treated trees sp ro u ted d u rin g th e first g ro w in g season. R esu lts a t th e en d o f th e secon d g ro w in g season are sh ow n in T a b le 1 as p ercen ts co rresp o n d in g to th e m ea n a n g le o f eq u a l in fo r m a tio n (arcsin \ J % tran sform ation ). T h e d ifferen ce b etw een 100 p ercen t to p -k ill for 4 0 lb a h g and 75 p ercen t fo r 20 lb a h g w as h ig h ly sig n ifica n t. T h e 64 p erc en t top-kill w ith 13.3 lb ah g w as sig­ n ifica n tly m o re effective th an 33 p ercent w ith th e 8 lb ahg. D if­ feren ces in c o m p le te k ill w ere sim ila rly sig n ifica n t. D ifferences in b o th to p -k ill a n d co m p lete k ill b etw een oaks a n d sw eetg u m w ere sign ifica n t, an in d ic a tio n th a t sw eetgu m w as harder to k ill than the oak sp ecies tested. T h e r e w ere n o sig n ifican t d ifferen ces in eith er top -k ill or co m p lete 13.3. a v is D ’Nacogdoches Research Center, maintained at Nacogdoches, Texas, coop­ eratively by Stephen F. Austin State College and Southern Forest Experiment Station,' Forest Service, U. S. Department of Agriculture. ’Manufactured by Reuel Little, Madill, Oklahoma. S .. k ill b etw een th e tw o tree s m trees sp ro u tin g (in c lu d in g t. greater for th e sm a lle r size T h e effectiven ess o f iso-oct oaks an d gu m s in ea st T e x c ester o f 2 ,4 ,5 - T ca n b e bas> W h ere o n ly oak s are to b e ci o f th e iso-octyl e ste r m a y be s sp ecies w ith sim ila r resistance co n cen tra tio n w ill be requir Lnx 1. B rasincto .v , J. J. Poisoning < Fanner 9(6):5. 1950. 2. D avis , J. R„ and D u k e . V . B . ( man 191(2393): 171-112. 1955. 3. C o p e , J. A., and S p a e t h , J. N . Jour. Forestry 29:775-783. 19' 4. G oddard , R. E. Rilling small ui tool. Down to Earth (Dow Ch 5. G rako , C. X . Effectiveness o f . Lumberman 185(2316):44. 46, 4 6. Jokela, J. J., and Lorenz. R. W. nating cull trees from woodlanu 1955. 7. Smith, J. L. Tests o f injected che sas mountains. Proc South. IV ¿v u 462 5293 * 3 463 G ibbs : I so -o c t y l E ster T a b le 1. Top-kill and complete kill two growing seasons after injection with 2,4,5-T. d Control ise o f a 36 lb ah g ester o f 2 ,4 ,5 - T ssen tia lly a m o d iid S p a e th (3) in fa m a te (1, 5) a n d V i % r i- s v a ila b le at lo w e r n itv l eth er ester, arch 1957 b y th e i th e Sam H o u so f th e ch em ica l, to 30 sw eetg u m s us stella ta W an chv.). T r ee s w ere : s. A to ta l o f ro u n d s, o n e p er effectiven ess o f sp ecial m easures o f th e s ilv id d e . and sp r o u tin g in en t, crow n d am • co n cen tra tio n s, ■ctive treatm en t, rees an d 93 per:etgu m than o n w in g season. ii are sh o w n in a n g le o f eq u a l :eren ce b etw een r 2 0 lb ah g w as lb a h g w as sig8 lb ahg. D if. D ifferen ces in sw eetg u m w er e to k ill than th e 2 O ak S w eetg u m B oth O ak S w e e tg u m B oth 4 0 .............................................. 2 0 .............................................. 13.3........................................... s .............................................. 100 92 69 39 100 54 55 27 100 75 64 33 100 82 64 35 100 50 54 23 100 67 59 29 k ill b etw e en th e tw o tree size-classes, th o u g h th e total p ercen ta g e o f trees sp r o u tin g (in c lu d in g those w ith liv e tops) w as sig n ifica n tly greater fo r th e sm a ller size class. T h e effectiven ess o f iso-octyl ester o f 2 ,4 ,5 - T w h en in jected in to oaks an d g u m s in east T e x a s m eans that ch o ice o f a lo w -v o la tile ester o f 2 ,4 ,5 - T can b e based o n rela tiv e costs o f th e ch em ica l. W h ere o n ly oaks are to b e co n tro lled , th e 20 lb a h g co n ce n tr a tio n o f th e iso-octyl ester m ay b e satisfactory. W h ere sw ee tg u m o r o th er sp ecies w ith sim ila r resistan ce to silv icid es are to b e treated, a h ig h e r c o n ce n tr a tio n w ill be req u ired . L iter a tu r e C ited S ' ¡Í :£. * .z. both* ?.sed, and# id. Table 5 -crs upon# 'he effect-4 u s to beii nd tissue^ discussed . ■on thy-J ated J ;? ? c • 1* - t í The iodinated amino acids of the rars. and in rats depleted of iodine by thyroid glands of treated and control means of the Remington diet for two rats were analyzed 24 hours after radio­ weeks there also was no effect of the drug iodine administration. Using the ion•m I :i uptake by the thyroid (Table 2). exchange chromatographic system of Since the association of an increased Galton and Pitt-Rivers (7) on pancreatin 1 thyroidal I 1*1 uptake with a lowered digests, we obtained the results shown in *rrum PBI is found in nephrosis, we Table 5. Total iodine analyses were per­ investigated this possibility, but kid­ formed on aliquots of the thyroxine frac­ ney histology remained normal, there tion and the pooled iodotyrosine eluates, was no increased urinary protein loss, and the percentage of thyroxine among and the serum electrophoretic pattern the total iodinated amino acids also was was unaffected by 2,4-D administra­ found to be unaffected by 2,4-D. The results of the determination of the tion. Iodide clearance, as judged by .•«rum inorganic iodide-Im levels 24 serum iodide level and PB I by the hours after tracer administration, also isotope equilibrium technique are shown ciid not differ significantly from the con­ in Table 6. trol levels. When we attempted to meas­ To confirm the magnitude of the serum ure other criteria of thyroid function, we iodide levels found, we calculated the f--und that many of these remained com­ difference between the serum total pletely unchanged (Table 1 ). Thus, the iodine and the serum protein-bound thyroid cell height and hiotslogic appear­ iodine levels in 11 control rats by chem­ ance were not affected by 2,4-D, and ical assay. This difference averaged neither was the rate of loss of radio- 1.0 ± 0.1 (standard error) /ig/100 ml, in i<>dine from the gland. When rats were good agreement with the levels cal­ kept on a goitrogenic diet for two weeks culated from radioactivity data in th e while being given 2,4-D daily, goiter animals in the experiment of Table 6 development was unaffected and the th at were on the same iodine intake. terminal value of the thyroid:serum radioiodide concentration (T:S) ratio was not elevated significantly (Table 3). T able 4. P itu ita ry an d serum th yrotropin assays i'ituitary TSH stores measured by Mc­ Kenzie’s assay (5) were unaffected by Pituitary TSH Content; 2.4-D. Serum TSH levels could be meas­ Controls (5) 93 ±15* mU per gland 2.4-D-Treated (5) 10S ±20 mU per gland ured only after concentrating pooled ■-*ra from 12 rats by Bates’s procedure Serum Levels; Controls 0.028 m U/cc 0 . We found levels of about 0.025 milli2.4- D-Trcated 0.024 mU/cc units 'mU)/cc of serum in both 2,4-Dt rested and control rats (Table 4). * Standard error of the mean. 5300 0003469 4 F L O R S H E IM A N D V E L C O F F Volum e 7j 4 T able 5. Analysis of thyroidal iodinated amino acids Controls 2,4-D-treated Number of rats 12 24-hr I:" uptake 6 .9 ± 0 .8 % Serum PB I 3 .9 ± 0 .1 0g / 10 O ml 24-hr conversion ratio 30 ± 2% I ” 1 Distribution: Thyroglobulin 4 .9 ± 0 .4 % loaotyrosines 62.6 ± 2 .4 % Iodothyronines 10 .6 ± 0 . 8 % I ,B Distribution: Iodothvronines as fraction of total iodinated amino acids 19.4 ± 0 .7 % 12 1 0 .2 ± 1 . 0 % 3 .2 ± 0 .1 #g'100 ml 26 ± 2 % Significance s of difference 4 (F) ; < .0 2 < .0 0 1 t : .' •2 4 .2 ±0.1% , 62.9 ± 1 .9 % 1 1 .7 + 0 .9 7 # 2 1 .0 ± 0 . 8 % -4 î > .i » :& -•> 1 i D iscussion control rats with regard to th e distribu-4 2,4-D has rather unusual effects on tion of stable iodine and of radioiodine 4 thyroid radioiodine uptake and serum among thyroglobulin, iodinated ty ro -i PBI, the two most commonly used sines, and tri- and tetra-iodothyronine.-4j criteria of thyroid function. I t is one of We did not determine whether or not the $ the very few compounds known th at iodide content differs, since this fraction * elevates I ’}1 uptake. The only previously was not separated from partial hydroly- -4 described situations in which similar re* ®s products and iodine bound to in-+t suits are obtained are nephrosis and soluble cellular debris. i iodine deficiency. In both of these there is The results of the 24*hour radioiodine * evidence of increased thyrotropin secre- uptake test, therefore, seem to conflict a tion from the pituitary, whereas we with a battery of other thyroid function 4 could show no evidence of this after tests. I t appears th a t the enhancement * 2,4-D administration. Pituitary and of thyroidal I >Muptake must be caused * serum thyrotropin assays support the by a change in the specific activity of f evidence afforded by the unchanged serum iodide due to a lesser dilution of thyroidal I m release rate and by the : the injected carrier-free radioiodide, i histologic appearance of the glands and The isotope equilibrium experiments at % the epithelial cell height. In our hands, both the 5 and the 10 fig per ra t per day 5 the index of precision of McKenzie’s iodine intake level showed clear depres- -» assay has averaged about .32, which sion of the serum iodide level. T he likeli- i allows for a large margin of uncertainty hood of radiation damage to the thyroid ■ in the results. With the low levels of TSH in these experiments is negligible, since i found in serum, which we estimate at the total radiation dose received by the i about 0.025 mU/cc, we could use only glands was well below the 3,000 r five mice per point, and this severely threshold below which radiation damage -3 limited the accuracy of the assay. The has not been demonstrated (10). The k TSH level we find in the serum of our iodide levels found were of the same A rats is much lower than th at which has order of magnitude as those calculated 4 been reported by D ’Angelo, using the from the difference between chemically \ stasis tadpole assay (9). determined serum total iodine and the i In the present study, we found no PBI, which is not in agreement with • differences between 2,4-D-treated and the work of Gerbaulet and collaborators * DOW 680485 j Ì .July, 1962 11 ), who ! scrum to 1 value calci 1*131 diffère derbaulet’. tains iodid iodide as tl pound m a\ It is of i isotope equ tarried oufrom radio: and signifie termined o: ¡cal analys The avera similar find Middleswo: this discret It does not to achieve the 26-day I’itt-Rivers calculating ing equalit; iodinated rr the tumov. iodine after that 23 da: 97l“c of eq tions, since 4.0 to 4.6 • Simon (15), technique i Experiment 1 : Strum Iodide l*Ul Chemical PB I Experiment 2: Scrum Iodide 1'BI Chemical PBI * Standar* E F F E C T S O F 2,4-D O N T H Y R O ID F U N C T IO N Volume 71 $1 raiacance '4 d:a*rence <.02 < .001 •t i .Ü * > .l ?» 3 listribu- 3 ioiodine -v i tyro- 4 Tonine. not the Taction ydrolyto in- •. i . who believe iodide levels in human ^•rum to be about one twentieth of the 7 :‘.ue calculated from the total iodinej>HI difference. However, it appears that (ie-rbaulet’s assumption th at saliva con* :.iins iodide in equilibrium with plasma ;.,.dide as the only iodine-containing compuund may not be valid (12,13). It is of some interest that, in all the isotope equilibrium experiments we have carried out, the PBI level calculated from radioactivity data was consistently .ir.d significantly lower than the PBI de­ termined on the same samples by cheroiva! analysis using TCA precipitation. The average difference was 35%. A -imilar finding has been reported by Van Middlesworth (14). The significance of this discrepancy is under investigation. It does not appear to be due to a failure to achieve isotopic equilibrium during the 26-day experimental period. Using Hit-Rivers and Hall’s (10) method of calculating the period needed for achiev­ ing equality of specific activity of all iodinated materials in the animals from the turnover rate of thyroidal radioiodine after a single tracer dose, we found that 23 days should suffice to achieve &:■ ; of equilibrium under our condi­ tions, since the half-life observed was 4.0 to 4.6 days. However, Morel and Simon 115), using a much more laborious technique for determining the attain­ ment of equilibrium, required 32 days to achieve 95% equilibration in rats th a t were considerably older than the ones used in Pitt-Rivers' and our studies. In experiments to be reported elsewhere, we have administered the long-lived isotope I ,Mfor 35 days and still observed highly significant differences between the PB I calculated from radioactivity data and the chemically determined PBI, which were of the same magnitude as those re­ corded in Table 6. For these reasons, we believe that isotopic equilibrium was , attained in our experiments and th a t other explanations must be sought for the discrepancies between the P B I values obtained by the alternate pro­ cedures. In any case, the determination of serum inorganic iodide levels, which was the object of these experiments, should certainly be valid since inorganic iodide equilibrates much more rapidly than the organically bound iodine. Thus 2,4-D appears to be one of the few agents other than iodide itself th a t has been shown to alter the serum iodide pool specifically. Our results again emphasize the necessity of using a large number of thyroid function tests when exploring thyroid-pituitary interrela­ tionships, since many peripheral factors, few of which have been explored ade­ quately, can invalidate several of the commonly used tests. 0 1 05 00 o ►Ö* 00 05 h w :2Æ?ïS? .3 f T able 6 . Isotope equilibrium determination of serum iodide and PB I Controls 2,4-D -treatcd Significance of difference (P) Experim ent l : 14 rats p e r group on d a ily done o f S tig iodide S trjjn Iodide P 3I Chemical P B I 0 53 ± 0.03 Mg/100 ml 0.78 ±0.03* pg/100 ml 1.92 ± 0 .1 0 pg /10 0 ml 3 .3 ± 0 .1 ¿ig/10 0 ml 0 87 ±0.08 >ig/100 ml 1 7 ± 0.1 Mg/100 ml 1.48 ± 0 .0 8 pg/100 mi 1.92 ± 0 .0 7 jig/10 0 ml 3 .2 ± 0 .1 ^g /10 0 ml 0 92 ±0.04 *g/100 ml 1 35 ±0.09 pg/lOO ml 1 9 ±0.1 «ig/100 ml Experim ent 2 : 14 ruts p e r group on d a ily dose o f 10 ng iodide .Serum Iodide PBI Chemical P B I ators .005 <.001 <.001 < .0 0 1 <.001 < .0 0 1 * Standard error of the mean. a 5302 0003471 F L O R S H E IM A N D V E L C O F F R efer en c e s 1. Sds, J . and P . K ertai, Acta Physiol. Acad. Sci. Hung. 14: 367, 1958. 2. D atia, S. K. and G. Banerjee, J . Indian Chtm. Soc. 3 1 : 397,1954. 3. Florsheira, W. H ., Endocrinology 62: 783, 1956. 4. Bodansky, O., R . S. Benua and G. Pen* nachia, Amer. J . Clin. Path. 30: 375, 1956. 5. McKenzie, J. M ., Endocrinology 63: 372, 1958. 6. Bates, R . W., M . M . Garrison and T . B. Howard, Endocrinology 65: 7,1959. 7. Gaiton, V. A. and R . P itt-R iven, Biochem. J . 7 2 : 310,1959. Volum e 71 ■ 8. Simon, C. and F . Morel, Int. J . Appi. Radial. 8: 35, 1960. 9. D'Angelo, S. A. and R. E . Tram o, Ann. N . Y. Acad. Sci. 72: 239,1958. 10. Pitt-Rivers, R. and J. E . Rail, Endocrinology 68 : 309, 1961. 11. Gerbaulet, K., W, Fittig and W. Maurer, Klin. Wschr. 38:474,1960. 12. Nicolas, G., Schweiz. Monatsschr. Zakn heilk. 70:633,1960. 13. Ceia, M., J . M. Sobral and W . Clode, Art}. Pat. 3 1 : 411, 1959. 14. Van Middlesworth, L. and A . P . Intoccia, Metabolism 6 :1 , 1957. 15. Morel, F . and C. Simon, C. R. Soc. Biol. 151:1106,1957. Iodide to Epir NORM / Harrison i University il -jj 41 o ? Oí 00 o CO <1 a| 11 j| H PH E : A berg, lease of thyrotro creased thyroid ■ re-exami from stu by the In these release ' thyroid ing adm epineph: mental i plasma vein anc these v; publishe were ini duced i examine port th< associat TSH. Ii . epineph prompt thyroid - M a le ; in w eigh Rcceiv * Prese Universi: M innesoi * Prese Stanford Alto, Cal 5303 0OO34|I2 -5*h Poes s Trans. 8b - f 12162 ftecord UOM 1 71415 MID. CODEN Cl Clinical Facts Lille Medical V. 7(10) 1049-51 (1962) P O L Y N E U R I T I S A F T E R U S I N G A N E E D KILLER: CONTAINING A C I D 2-4-D. By Philippe Foissac - Gegoux, Annie Lelievre, Bernard Basin and Pierre Warot. B y 1958, one of us (*) had already drawn attention on the risks Involved wit h the m o r e and the mor e frequent use of chemi­ cal p r o d u c t s in agriculture: two observations were considered at the b e g i n n i n g of this w o r k which concerned the dangers de­ rived from handling organics derived from phosphorus, powerful i n s e c t i c i d e s ; one of these observations concerned an agricul­ tural w o r k e r who handled, m a n y months before the installation of a p o l y r a d i c u l o r e p h r i t i s syndrome weed killers, sodium and °003930 5305 (L p o t a s s i u m salts of 2- m e t h y l -4-.c hlorophenoxyacetic acid; f o r several reasons, we had eliminated in this case the toxic origin of the neurological troubles..., but we ignored at that time ÜÜW 1 714156 that these products could have a toxic effect. In truth, the cases of intoxication due to weed killers are rare and are e s p e cially known by toxicologists and doctors; but it is feared that they m a y multiply: for almost all.the substances destined to stop the development of weeds are d an­ gerous to man. M i n e r a l comp o u n d s were being used such as chlo­ rates and sulphuric acid: these were three times more incon­ venient wit h an action often little selective, hence a global destruction of grasses and cereals, of a delicate use (risks of explos i o n and fire) and of a sure toxicity (burns-methemoglobinemy). The "vegetable hormones", an und i sputed progress. the last commers, make up This name is gi v e n to chemical bodies whose action is comparable to that of phytohormones elaborated by the plant itself. In small doses, they in fact stimulate the growth, of y o u n g v egetable cells; b u t in the large doses that they are used they cause "a crazy exaltation o f the m a n i ­ festat i o n s of growth, a dis o r d e r l y a c t i v i t y which drives the m o s t deverse monstruositles, the intense Consumption of reserves and f i n a l l y the death o f the grass". Uiese "hormones" are oxyacetic derivations of the benzine nucleu s and of the n a p h talene nucleus of the phenyl, naphtyl and indolacetic, propionic and butylic acids as w e l l as some of th e i r derivatives. One of the m o s t wide l y used is 2-*J- dichlor o p h e n o x y a c e t i c acid (Ac. 2-^-D.) whose action powerful a n d 5306 i' n '■ 'i 1 selective, Is not, however, w ithout Inconvenience as the following o b s e r v a t i o n proves. Observation: G . .. Gerard, 52 ye a r s old, farmer, was referred to the N e urol o g y Clinic on J u l y 30, 1962 by D o c t o r Chuffart for sen­ sory and m o t o r p r o blems of the lower limbs. This man, without any pr e v i o u s p a t h o l o g i c a l problem, w h o Is not an alcoholic, dates the b e g i n n i n g of his troubles prec i s e l y on June 2, 19 6 2 . On that date he felt, suddenly, an acute p ain throughout the entire right orbital and perio r b i t a l region becoming weaker the next day m a k i n g roo m for (burning sensation) p a resthesra and numbness in the same area. A l s o on June 3 rd , he noticed a (decreased sensitivity) h y p o e s t h e s i a in the right leg. On J une 16, two weeks.later, the right leg recovered a normal s e n s i t i v i t y but the n u m bness was now felt on the left leg; besides, the l o w e r left limb is n o w "weaker" and cannot move e x c e p t w i t h the aid of a cane. The p r o b l e m s r e m a i n e d about the same untlll J u l y 30 when he entered the service. He n o w felt a persistant hypoesthesia in the ri g h t eye area, w i t h h y p o e t h e s i a of the cornea, the patient now felt some p a i n in the same area. The s u b j e c t is u n c e r t a i n of his lo w e r left limb where a alight r e d u c t i o n - g l o b a l - o f the segmentary force is noticed. There is no o t h e r trophic va s o - m o t o r trouble. The knee r e f l e x e s are r a ther sharp while the ankle reflexes are suppressed. The l o w e r left limb suffers from a global 5 3 0 7 GC0^ 3 2 h y p o e t h e s i a o n l y deep s e n s i t i v i t y w a s maintained. The n e u r o logical ex a m i n a t i o n of the upper limbs is strictly negative. Besides the g e n e r a l cond i t i o n is excellent. The blood f o r m u l a reveals an eosinophilia (8# o n A u g 2 and 9# Aug 30). Two electrophoretic blood tests, several weeks apart show a slight decrease in a l b u m i n a (43 then 46 grams) and- a n increase of a l p h a - 2 - globulines (1 3 then 15 grams p e r liter). The renal bi o l o g i c a l exam, DOW 1 714158 is no albuminuria. There the galactosuria provoked, the reactions of f l oculatlon do n o t show any abnormalities. The e l e c t r o c a r d i o g r a m is normal. the eye and the E.E.G. results: So is the b a c k s i d e of The e l e c t r o diagnosis shows the following ------ p r o o f of s t i m u l a t i o n of Aug 2, 1962: of the u p p e r limbs; Integrity in the l o w e r limbs there.is no evident weakness but slight problems of e x c i t a b i l i t y (galvanic hyperexcitability galvanic and faradic h y p o e x c i t a b i l i t y of the e x t ensor of the left b i g toe ----- h igh c h r o n a x l e s of the extensors o f the left big toe) w itness h o w e v e r a slight neurogenic seizure (Doctor Spy); ------- e l e c t r o m y o g r a p h y of the two frontal tibia on A u g u s t 4, 1962 (Doctor Ramez): trace of ne u r o g e n i c seizure of the two muscles examined. The p a t i e n t leaves the c l inic on September 8 , 1962: during hosp i t a l i z a t i o n he was subm i t t e d to a daily injection o f 200mg of v i t a m i n B1 and 250mg. of v i t a m i n B 6 ; besides 5 > 0 0 0 gammes of hydroc o b a l a m i n e were a d m i n i s t e r e d every three days. aches p r a c t i c a l l y disappeared; T h e facial the deficiency of sensitivity in 5308°° the face and the lower left limb was notably Improved* the m o t o r d e f i c i e n c y disappeared and the patient was able to wal k normally; the a reflexia achillelne persists, there is no v i s i ­ DOW 1 714159 ble atrophy. There are still el e c t romyographic signs of neurogenic seizure of the right frontal tibia (the left side was not examined). On the c o n t r a r y the electrodiagnostic of sti mula­ tion :of A u g u s t 28 are a b s o lutely normal. The a e t i o l o g y of this p o l y n euritis of the lower limbs with ir r i t a t i v e and d eficient right trigeminus was suggested to us b y the pat i e n t hi m s e l f who was able to relate the be g i n ­ ning of his trouble while killing grass. the last days of M a y 19 6 2 . Made June 1st and This o peration consisted of a spraying o f two different solutions but b oth having a base of acid 2 - 4 -D, c o n t a i n i n g 235 and 410 gr a m s per liters. It was carried out w i t h a t ractor wh i c h tows the spraying device and which is in a cabin, open in the back; it was very w i n d y when the o p e r a t i o n s took place and the wind beat the toxic substance in the cabin. These e x c e p t i o n a l circumstances, the long exposure to the toxic substance, we had to a d m i t that it was a p o l y neuritis due to 2 - 4 - D acid. B e s ides, this o b s e r v a t i o n is not alone in the litterature that i n c l u d e s a c e r t a i n n u m b e r of facts o f intoxication by these substances, n o t a b l y in the case o f farmers: some hours after inh a l a t i o n of the toxic substance there is generally vomiting, 5309 c c O *J !j *1 6 then subjective sensitive troubles of, the type, o f .paresthesis o r e v e n pain, then m o t o r troubles, limbs o r the l o w e r limbs. This sensitive - m o t o r polyneuritis i n c o nstantly associated with an albuminuria and an inter- ~ o c o l i t e board pic t u r e chart description ------ is usually irre­ versib l e or very slowly and ver y p artially reversible. The toxicity o f these substances is equally proven in the case o f intoxication of w orkers at a vegetable hormone factory: Die p e n e t r a t i o n of the toxic substance through inhalation, but also through the skin; the workers often complained of somnolence-, of the l o w e r limbs feeling heavy, gastralgia, loss of appetite, hypersialosis, as well as hypers e n s i t i v i t y of hearing: these troubles are, generally, transient. all of Finally, experimen­ tally, 2 - 4 - D acid administered intravenously or even orally can be fatal: thi3 happens suddenly, either in a neurological stage in the form of a c o m a wit h hypertonic of the limbs, or ventricular f i b r i l l a t i o n w i t h relatively large doses, but varying according to the animal tested (about 3C0mg to one gram). To our knowledge, there w a s no acute intoxication. Bibliography A s s o u l y M . - Selective weed killers and grow t h substances. T e c n i c a l survey. Path o l o g i c a l effect on m a n during the m a n u ­ f a c t u r e of the e s t e r of 2-4-D. Arch. M a i . Prof., 1951 1, 2 6 . G o l s t e i n N. P . J o n e s P.H. and Brown J.R. --- P o l y n e u r i t i s a f t e r exposure to an ester of 2 - ^ - D acid. DOW 1 7141G0 ' the paralysis of the four I * * H i l l E.V. a n d C a r l i s l e A. ------ T o x i u i t y o i 2-*l-D. for e x p e r i m e n t a l animals. Jour. Indust. Hyg. and Toxic, 197^, 29, 2, 85-95. O.R.S.T.O.M., Paris, 1958. Footnote p. 1 (*) P. Nayrac, M. Fontan, J. Delahousse: P. Warot, J. Lescut and L i l l e Medical, DOW 1 714161 Lhoste I. - C h e m i c a l weed killers. 1958, 3* 3» 161-16U. 5311 0C0333G t ó - n - iy 9t ¿ « 1 7 5 cu ra i’} : ■ i-IU£ M B b lC A L / o f f . j - / ( n c r ï) l'O L Y X K X 7.77'/; A P I t K s rsA U K D I X m i s //A7.7M.\T : L '. U 'I D K par l'IiilipiN* KiiL'%'.»r-»*'K«airx Annie L i-3.ikykk. l!<-ni;inl M\six et l’iorre Wtittvr. A v ra i d ire , les ea s d 'in to x ie a tio u p ur d e s d ésh erb a n ts so n t rares et son t su r to u t co n n u s de» to x ico lo g u e s et d e s m éd ecin s d u tr a v a il : m a is il est à c r a in d r e q u ’il» n e se m u ltip lie n t : c a r p resq u e touti > les su b sta n c e s d estin ées à e n tr a v e r le d é v e ­ lop p em en t d e s m a u v a ises horl>es son t d a n g e reu se s l>onr l'h om m e. O n u tilisa it n a g u è re de» com p osés m in érau x p a rm i le sq u e ls les ch lo r a te s e t l'a c id e su lfu r iq u e : c e u x -c i a v a ie n t le tr ip le in c o n v é n ie n t d ’u n e a c tio n s o u v e n t jm-u sé le c tiv e , d 'o ù u n e d es­ tru ctio n g lo b a le d e s h erb es et d es céréa les, d ’u n em p loi d é lic a t (r isq u es d 'e x p lo sio n e t d 'in c e n d ie ) ■-( d 'u n e to x ie ir é c e r ta in e (b rû lu res • m éth ém o g lo b i­ n ém ie). I.es « h o rm o n es vég éta it1» », d ern ière» ven u es, (» in stitu e n t u n p ro g rès in d iscu ta b le . Ou d o n n e ce n om à d e s co rp s ch im iq u e s d o n t l'a c tio n est co m p a ra b le à (»die d es p h ytoh orm on e» é la lx n c c s pur la p la n te elle-m êm e. A d oses fa ib les, e lle s stim u ­ lent en effe t la cr o issa n c e d es c e llu le s vég éta le» je u n e s ; m ai» a u x d ose» forte» a u x q u e lle s on le» em ploie, e lle s p ro v o q u en t « u n e e x a lta tio n fo lle d es m a n ife sta tio n s d e cr o issa n ce , n u e a c tiv ité d éso r­ d on n ée q u i e n tr a în e l'a p p a r itio n d e m o n stru o sités les p lu s d iv e r se s, la con so m m a tio n in te n se d es réserves et fin a lem en t la m ort a u s ta d e h erb acé ». (*) P. N avra«', i!. F oxta». P. W arot. J. L khiut et J. Du . vkocssk : /.;/!«• M i-tlicul. 1958. X 3. 161-164. t'e s « h o rm o n e s » sont des d ériv és oxyncétiqucs d u noyau benzène et d u n o y au n ap b taiciic des acides p bcnyl, n n p h ly i et iiidolucétiquc. propion iq u e et h u ty liq iic. a in si q u e d e c e rta in s de (cuis d ériv és. 1, ' o.î ; des p lu s em ployés est l'a c id e 2-4d iclilo ro p h én o x y acétiq u c (A c. "2-4-D.i dont l'.iction, p u issa n te et sélective, n ’est c e p e n d a n t pas •léitucc d e tout inconvénient ain si q u e le pro u v e l'o b serv a­ tion su iv an te. DOW 1 714162 - D é j à «-ii lî»r*S. I*un d 'e n tr e n ou s t.*) a v a it a ttir é l ’a tte n tio n -sur le s risqu e» q u e com p o rta it l 'u tili­ sation d e p lu» e n p lu» fr e q u e n te en a g r ic u ltu r e d e p rod u it» ch im iq u e» : d e u x o b ser v a tio n s éta ie n t à l'o r ig in e d e c e tr a v a il d a n s leiiu el l ’a ce cn t a v a it été m l» s u r le» d a n g e r s do la m a n ip u la tio n d e s d ériv és o r g a n iq u e s d u p h osp h ore, in secticid e» p u is­ sa n ts ; l'u n e d e c e s o b ser v a tio n s co n cern a it un o u v rie r a g r ic o le a y a n t m a n ip u lé, p lu sie u r s m ois avan t r in s t a lla t io u d*un sy n d r o m e d e p o ly ru d icu lo n évrite. d es d é sh e r b a n ts, s e ls so d iq u e et p o ta ssiq u e de l'u eid c 2 -m éth yl-4-oh loro«ih én oxyacétiq u o ; p o u r p lu sieu rs raison», n o u s a v io n s é lim in é d a n s eu ea s I o r ig in e to x iq u e de» tro u b les n eu rologiq u e» ..., m a is n ous iu n o rio n s à l'é p o q u e «pie ces p r o d u its p o u ­ vaient a v o ir u n e a ctio n to x iq u e. *.’r /h O B S E R V A T IO N : 0 „ Cuvant. 52 ans. agriculteur, est adressé à U (.'Unique Neurologique te 30 juillet 1962. par 1« Docteur C n rrro tT pour des rrnuldt-s senxitifs ;•( moteurs de* memlnv.s inférieurs. Cet homme. sans anti-relents put hologiques notables, qui n'est pas éthylique, situe très précisément le début de ses troubles au 2 Juin 1962. H ressentit ce jour-ln. brusquement. une douleur vive duns toute la région orbitaire et périorbitaire droite, s'atténuant dès le leiulemain (tour taire place 4 des paresthésies et à une a-.n-stiiésie ncctqiaiit le mén:>- t -rritolre. la* 2 juin également, il note l'apparition d'une Itvpoestiiésie de la jambe d ro ite. Ia> 16 juin, soit deux semaines aînés le début. la jambe droite a récupéré une sensibilité nor: ale mais l'anesthésie, qui persirto & lit face, a gagné la jambe gauche ; d'autre part. le membre inférieur gauche est - plus faible a et la marche nVst possible qu'avec l'aide d’une canne. Los troubles restent sensiblement identiques Jusqu'au 30 juiller. date de l'entrée dans te service. On constate alors la ix-rsistaner d'une liypovsiliésie dans le terr.toire de (’ophtalmique droit, avee hytioesrliésie mrnéeunc ; le malade accu..>as d 'a lb u m in u rie . .«angtiine ré v é le u n e é o s in o p h ilie . sa n g u in s . & p lu s ie u r s s e m a in e s d 'in t e r v a lle . mont t i nt lég ère de l'a lb u m in e (13 put.« 46 tin»* hait*»* g ra m m e s) une h p a r lit r e ) . le s ré a c tio n s il« iln ciila tin ti n e m o n tran t p as d 'an o m alie *. L ’é le ctro ca rrilo R ra n im e fond psi m e m b res lé g e rs s u p é r ie u r s ; tro u ille s g a lv a n iq u e de aux m em b res l'e x c ita b ilité • h y p o e x c ita b ilité in fé r ie u r s , il (h y p e r e x c ita b ilité g a lv a n iq u e et fa ra d iq u e île l'e x te n se u r p ro p re du R ro s o rte il g au ch e - e b ro n u x ie s ce p en d an t d 'une létcère a tte in te ueurogènc (D o c te u r t j r x ) : — é ie rtro m y o g m p h ie des d eu x ja m b iè re a n t é r ie u r s du 4 a o û t 1963 ( D o r t e u r I I a m i x ) : tra cé d 'a tte in te neuroRêne di*a d eu x m u s c le s e x p lo ré s. m a la d e q u itte la c lin iq u e le S sep tem b re 1963 : d u ra n t l'h o s p ita lis a tio n il a été s o u m is à u n e in je c tio n q u o tid ien n e de 2nn niK. de v ita m in e B 6 ; en B1 et 2.'0 m g.. de o u tre .' l'tiy d ro co b a la m ln e lu i a été a d m in is t ré e & la dose de 5.ut«t R a n im a s tous les* tr o is jo u r s . Les a lg ie s f a c ia le s h a la t io n il u to x iq u e s u r v i e n n e n t lia h it iu ili-m o n i île s v o m is s e m e n t s , p u is «1rs t r o u b le s s e n s it i f s s u b j e c t i f s OH à t y p « tle p a r e s t h é s ie s o u m é m o «h* i lo u le u r s . p u is é lev é e s d es e xten si tirs du s r o s o rte il c a n d ie ) téntoiRnent v ita m in e tn m il.ro «le île s donne n’y a (tas de r é a c tio n de ra le n tisse m e n t é v id e n te m a is Le c e r ta in L 'é le c ir m lia g n o s tic — é p re u v e dp s tim u la tio n du 3 août 1!*62 : in té g rité le s un p a r e c s s u b s t a n c e s , n o ta m m e n t n o rm a l. 11 pn pat de m êm e d V cii et dp l ' E .E .G . le s re n se ig n e m e n ts s u iv a n t s : dp* f a i t s d 'in t o x ic a t i o n e lle * d e s a g r i c u l t e u r s : ip n -lip p -s h e u r e s « p r i s l ' i n ­ L f b ila n b ioln iiiq ui1 r é n a l, la u aln rtoxiirn - pr«»v«H|uée, du l i t t é r a t u r e ip ti m m p o r t o ont p ratiq u em e n t d is p a r u ; les tro u b le s s e n s it ifs d é fic ita ire s de la face et du m em b re in f é r ie u r g a u ch e se so n t n o tab lem en t a tté n u é s ; le déficit m o te u r a d is p a r u et la m a rc h e est red even u e n o rm a le ; P a ré fle x ie a c h illé e n n e iie rs is te ; a u cu n e a tro p h ie ne s'est d essin é e . 11 p e rs is t e d es s ig n e s é le ctro m y o g ra p h iq u e s d’a tte in te n eu ro g è n e d u ja m h ie r a n t é r ie u r d ro it (l'e x a m e n n 'a p as àitm au g m e n ta tio n «les alphu-2-glohulint-a (13 p u is l à g ra m m e s A i t re s te , e e it e n lis i- r v a t iu t i n Y -d p u s iso lé e iln u s la t r o u b le s m o te u rs, l'a t t e in t « ' p n r a ly t j t p i c jm>u - v a n t i n t é r e s s e r b-s « p ia tro in i-n ih rc x n u h 's m em bre;« ^ in f é r ie u r s . — ^ in r p n s t u n im c n t nssoeiéi* à un«* n ib u m i n u r i e «■( à u n ç t a b le a u ^ iV t t c |M ilynévrit«* d 'e n t é r o c o lit e — M - n s itiv o - n io trie e s«- r é v è le h a b it u e lle m e n t i r r é v e r s i b l e «,u t r è s le n t e m e n t e t t r è s p a r t ie lle m e n t r é v e r s ib le . I.a itixieifé «1«* «•«•s stilista n ees est égalem ent prouvés' par b-s cas «l'in to x ica tio n ch ez le s ouvriers ■ra v a illa n t à la fab rication «les b orm oues végétales: la |MMiétruti»M «lu to x h tu e s«' ferait p ar iubalati'in. m ais au ssi par v«»i«* «-ulanéc ; b-s o u v rie rs se plai­ g n en t sou ven t «le som n olen ce, tle se n sa tio n s «le l«»ur«l«*ur M .K. and N««i:im.\ ll.T. — Effe«-ts of 2-l-«lich!«’ -i. p!n-no.\yai:**ilc acid on chicks a‘ci> 194$. M«>.v «s « a <; c i D i V i n i C . — S u r l'in to x ic a tio n alxui* p a r un d fe ii'-rb a n t. 1'aciile 2-l-D . C o n trib u tio n ctlu iq u e. f ' t l i ’i M n lic tt. 1361. | | . 6. :S0-!S5. i73-4S'i. Les «-mpto-s. <•« ucricuitOTA d*-s désbrrbanlT sélicilis • l ui» végétation. A y ric u lttm - Ti*.\ Al.S. — -Ac -suj*-t »it** «¡«^herbütm r«;lectifs. A n h. X'tl. 1951. 1. 2C-d. «t lu. ,i M oxorym uE M :* lad ilte . de rx cas de ai e m s d s Pierre W’akot. -bn-ipies IH x v x . René Lj-xuik et Michel D uuuiihs . L on gtem p s con sid érée com m e l ’a p a n a g e p resqu e e x c lu s if «le l'a n im a l, Jr in fe c tio n listé ric n n o sem b le m* m a n ife ste r ch ez ("homme a v ec u n e fréq u en ce rue : d'après Bi_VA^yr et SnuiEU, TO cas seulenient «le listériose lutmuVnc avaient été rapportes avant tandis «pu* .‘liai nouveaux eus l'étaient »K* lilâti à lilâT. \ Sans «lotit«- I«* «ruiun«>stic\ bactériologique est-il fait plus souvent «pt'autrcfolsY « o u passe souvent à «*ôté «lu diagnostic ImetérioIieaSque paire que l'on «Varie trop vite. s«»us l'ap p ellatiu V d c facilité de « eoutaiiiiiiants diphtériniorphcs » ilc-Ttau-illes ('¡nuit positif is««lés d'hetiiocitltures, «le liqUiHe céphaloraeliidien ou «le tout autiv produit e t \ i u i vint •l’autlienthiues listét ia » ( L . u ' i : y w o x s i k ) . \ l a i s il est très vraisetulilalile aussi qui- la diffusion, \eeim nue, «le nuf.-eij.iti dans le règne animal oiilfluîne la niulliplieatioti «les transmissions acvidentellcai à l'homme. Il est rare «pie le e liu ie ie u fa sse le d ia g n o stic «le listé rio se i-t en d em a n d e eo n lir m a tio n a u bio­ lo g iste : rien «l'étonn ant à c e la si l'o n co n sid ère q u 'il s 'a g it .1'u n e in fe c tio u d e c o n n a issa n ce récen te e t d on t la fr é q u en ce e st d a ille u r s to u te r e la t iv e : la litté r a tu r e fr a n ç a ise n e eo m p ortait en effet, en 19."f). u u e i l o b serv a tio n s. D 'a u tr e p art, les listérios«-s se p résen te n t so u s d es a sp e c ts e |in iq u e s e x tir m e m e n t «livcts : l'im p o r ta m e sta tistû p ie «le S i:i.ini:kii. [ x n ta n t s u r les .‘144 ea s p u b lic s «laits le inonde en l!)û(î. relève USD é ta ts scp tiru p yoh cm iip n -s «lu iiiiuvi-au-ué. 100 m cn iiig itv s p u r u lc u le s avis* ou sa n s e n c é p h a lite , 41 form es sa n g u in e s à ty p e «le inonotiui-léose iiife e tie tise . Kl e o n j u m tiv ite s g r a n u ­ lo m ateu ses. le s a u tr e s «-as i*««neeruuut «les lo ca li­ sa tio n s v isc ér a les «liverv-s : <-iulo«*ai‘d lte s. m yoear«lit«-s. liu 'lrites, «-le... L il l e .Médical. î Tout au plus pi'ut-ou parfois soupçonner l ’ori* gin«* listcrieune d'un état septicopyohéini«iuc du nouveau-né. ou «l'une méningite purulente de IVnrant «m de l ’adulte, parce qu'il s'ugit là des manifestations les plus frequentes de l'infection. .Mais, le plus souvent, le mérite du «liagnostie revient au seul baetériologistc... et ce n'est «pl'a posieriori, lorstpie l«> germe est identifié, «jue le elinieien mettra l'accent sur «(uel.ptes nuances séméii>l«>gi The noxious effects of 2,4-dichIorophenoxy-acetic add have been under detailed study at our Institute [5, 6, 13, 14], in view of their significance in labour hygiene. Dichlorophenoxy-acetie add (in the following DCPA) is a «ted killer used in increasing amounts mainly in corn fields. It has been shown by us th at chronic treatm ent with DCPA caused very dow, 2 to 3 Ha, high amplitude waves to appear on the electroencephalogram, and poisoning with the substance induced disturbances [S, 6] in. thyroid fraction [13]. As changes in thyroid function are known to produce eleetroeneephalo* paphic changes [8], the question arose whether DCPA damaged the nervous ayitem directly, or the changes observed in the EEG were due to an action •a thyroid function. This problem has been investigated in the present experiments. A tu tal of 15 c ita , 11 esperimenti! sud 4 contro!*, weighiag L S t * U k |i «ero a n d , ffcs ttectricsl aedvity of thè centrai aervous System was studied la eneh animai by maona ri 2 frontal aad 2 oceipital «orticai built-ia eketrodee, as watt aa bipolar sabaortieul abetrodee yrissd hi thè maaoacephattc rotieular formatimi. To placo thè eortkal «locaoda*, opeaàags { s a in diaaMter had beau cut la thècalvarium and tho stivar alactrodas w on ploeod on tao j^ a . To iasart thè subcordcal eleetrodaa, aa opaning 4 mm la diamatar was mède, thè dura m inristifi tho silver eloetrodes aa d asod in a class tuba wero iaaertod iute tho rotieular fori by maana of a sterootaetle apparata*. The coordinatee were detarudaad ocoorttag to gs—vlcornAi [9] A itar i : tho' riectrodss a fibria sponge waa placed la the drill boia, ■¡I* haarittr of the deep was eontroQed' hiatolagiraily aitar tba experiment. •Si ¡1 ** i 35' 74 L DtU, J. SdS saS L 1UK0LR3 Tk» «Iwtw J ii v m w w t r i in place w ith acrylate. A fter EEC tracings had basn recorded from the otherwise intact cats, they wets jeeted to thyroidectom y under aseptic conditions. Sahsaqnantly, the controls wars not treats* in any way. The experim ental »■«»»«1« wore given 100 pg/kg thyroxine (Hoffm an*—La Rack*! daily. Wo than stadiad for 10 days tha experim ental group to determine w hether changes a the EEC would occur. No appreciable changes having been noted, from the 11th day oa u* experim ental animals were given 100 mg/kg of DCPA daily. Tha m aterial was obtained aft** recrystallising three tim es the commercial preparation. Using a 4-channel EEC apparatus, tha spoutaneona electrical activity of tha eoetss and reticular form ation was m ooned daily. We also stadiad the changes in tha duration of cortical desynchronisation in rsspesw to th o electrical stim ulation of the reticular form ation. Tho apparatus need for stim ulation em itted rectangular impulses, w ith tha paramstn* « V and 1-5 V, frequency 300 Ha, duration 3 msec. T otal duration of stim ulation was 5 ***• Considering th a t tho stim ulations w en applied once a day, no adaptation could infleeo* th a resp onses according to d ata in tha literature [2, 4]. The cardiac rata was determined daily. Result» < iw The results. obtained m ay be outlined aa follows. In the control groop th e pro-thyroidectomy avenge frequency of 6 to 7 Ha decreased 24 hoot* after thyroidectomy and remained a t a low level throughout. In tho experimental group no appreciable change in frequency ws* noted after thyroidectomy. On the 10th day after thyroidectomy the frequency ranged from 5-5 to i i H i, aa compared with the pre-thyroidectomy value* of 6 to 7 Ha. On the other hand, frequency decreased 24 hours following the administra­ tion of DCPA. and after 5 days it was as low aa 2 or 3 Ha (Fig. 1). .13 « »1 Fig. 2. Changes in the frequency of spontaneous cerebral electrical activity in the eontrd and the experim ental group Arrow: thyreideetom y. Double arrow: onset of DCPA treatm ent 5317 0006317 •• momna s n or action or a num i 75 Similar change* were in the amplitude of the wave* (Fig. 2). In the control group the 60 to 70 fiV amplitude* slightly iacreasedafter thyroidectomy, then remained a t the 90 to 100 /tV level throughout. n n » - U M ,J |t In the experimental animal* the initial value* were the tame and 10 day* ether cbaagM t»r1 efttr thyroidectomy they were «till at the 60 ftV level. 11th dmy »ho. ,(obtained titer, ihey were mb* Mata not treats* DOW 1 249364 Uthe peremeten ilation wee 5 tee. •a could inflneaee < control group' sated 24 hour«* » frequency tn t] f the frequency a lectomy valuesj Fit. 2. Change* ia tba am plitude of «pontaneoua earahral tU ctrieal a c tin tr, thetdminiatra«^ Fig. 1). Sign* «a ia Fig. 1 O rdiaata: am plitude ia /»V. Ahada«a: day* Twenty-four hour* after the onset of DCPA treatment the amplitude hneoed parallely with the decrease of frequency. On the third day the average amplitude was 100 /jV, and by the fifth day it reached value* around 180 la 190 In the ££G tracings of the controls (Fig. 3), 3 days after thyroidectomy lin t was a significant slowing down (Fig. 3/II), as related to the initial value {fig, 3/1). The rhythm was still slower 11 disys (Fig. 3/111) and 15 days (Fig. jfpT} following thyroidectomy. l( In the experimental animal*- (Fig. 4) there was practically no EEG , ¿jnge after thyroidectomy (Fig. 4/II), as compared with the pre-thyroidectomy : Bsdag* (Fig. 4/1). Twenty-four hours following the onset of DCPAadministra* : jjgg frequency decreased and the amplitude increased a t sites (Fig. 4/IU ). | ¿flax 5 days the tracings displayed exclusively big, slow,- toxic wave* (Fig. 4/IV). tivity ia the etinent The duration of desynchronisation in response to stimulation of the formation showed characteristic changes (Fig. 5, Fig. 6); the value |(gfrmmd before thyroidectomy was taken to he 100 per cent, later deviations item calculated on th a t basis. ,0006318 , 5318 7« x. otu. j. ads a t l mxoLiTs la the control group the duration of desynchronisation rapidly decreased after thyroidectomy» From the 6th i i y on no desynchronization resulted ■ response to stimulation with 0.2 Y. From the first day on the duration of desynchronisation elicited by stimulation with 1.5 V was markedly reduced* lX u m ss) as fi 2-7 7-0 m s -9 /w F{f. 3. EEC r f control gronp. Diagram: cita and eonnaxion batwcan dectrodas. Nuiw—b* No. of rà g ia dectrodas. FR : reticolar form ation load L P rio r to thyroideetom y, aorm ai a ctirity . IL 3 days a itar thyroideetomy. Storia* Increata e f ampUtode, dyirhythm ic corra. IO . 11 days aitar thyroidoctomy, and IV. » days a ita r thyroideetom y: slowiag and dysrhythm ia slightly iacreaaad In the experimental group desynchronisation time was reduced by 16 per cent on stimulation a t 0 2 Y 10 days after thyroidectomy. The duration «m desynchronisation decreased rapidly on DCPA administration, after 24 hours by 50 per cent already, and on the fifth day no desynchronisation at all resulted on stimulation at 0.2 Y. »E nvois s m o r Action o r a chkmical h cubicioè r 77 Tea days after thyroidectomy the desynchronisation elicited by 1.5 V ^imnUtioa was reduced by 13 per cent. The duration ,of desynchronisation rspidly decreased in response to DCPA, to 45 per.*ent-by the Sth day. DOW 1 249366 7-3 Ffta-a 3 -2 B 2- 7 7-3 FBB-9 t—/****—*. wAAAwyVAsn % „i»V y , >»n 3- tv 2- 2 7 7 -3 ra e -a 3- 2 Fig . 4. EEG of experim ental prm p. Si$ns u in F lf. 3 r N otasi activity before thyroidectom y. IL Normal activity 10 days after thyroidectom y in 24 boars after onset of DCPA treatm ent. Slowing and a t sites increase of am plitude. [Y, 5 days after onset of DCPA treatm ent. Exdnaively big, alow, toxic delta w ares are risible Changes have been noted in the cardiac rate, too (Fig. 7). In tbe control poop thyroidectomy waa followed by a gradual decrease, in the experimental paap the heart rate decreased by 5 per cent in 10 days after thyroidectomy, following the onset of DCPA treatm ent the decrease was 25 per cent after the Hist day and on the 5th day it was 36 per cent. C006S2.0 : 5320 K 78 l o t s . u d i a i i m xou n Accord; ^Identical in t i(n d ia anixna j.' The apf ■definitely in ^together witi « marked \ \ * \ ■ttpatmmM 02 V •m ini | f ? J * 5 « 7 J 0 Fig. S. of tk * 100 M 02V 13V 13V JOjjff 12 » H IS C k tu it la tlM do tation o f to r d o l daayaehroaisatioa a f a r «lactrieal adwdrd** M k u v form ation. Graphical representation. D eration M a n thyroid******* se a t. Dtffaranca from this g trsa la par eant. Arrow: thyroidectomy. D"** arrow: oaaot of 0C ?A traatm ant r : 1m 1X)* I |J h .« .'H 'I’ll, i.'i r *]■ i Fig . 6. Changes la th s deration of w r i n l deaynchroniaatioa foOowiag adm olatioa of tb* rooealar - fbrm atioa : w ith 13 V. Arrow: tho C ut, low-ampiitnde deaynehroonati** aoaaaa aad tho baaal aetM ty reappear». ......... L : Experim ental groap. II: Control groan. 1/1, _ J„. M o n thyroidectom y. 1/2: Ton day* a f a r thyroidectom y. Doaynchum liatioa tim e hardly ahortaaod. 1/3: S days a fa r aaaat » DCPA troatm eat. D uration of dm yachraaiiation strongly reduced. H/2: $ day*, n/3 1» day*, a f a r thyroidaetom y. Darmtioa of daayachreaiaatloa strongly radacad 0006521 5321 Compa the dixninnti In the ■lowing of fro ‘ iJ w tr fit we • literatur During ht the ex per l |. The cfa treatm ent pi Thu», i with intact The ch M severe a s ’ nacrous m or action or a cbodcal a a m ia s s 79 D iscussion According to the above data, the EEG cbangea induced by DCFA w en iitnticsl in thyroidectomised animals maintained with thyroid sobstitationi in -"■"»■1« with intact thyroid [5, 6]. The appearance of 2 to 3 H*, over 100 ftV, big, alow waves (which are definitely in the pathological zone) is indicative of a severe toxic state, and together with the shortening of the duration of desynchronisation it points is a marked reduction of central nervous excitability. D o n 249368 Comparison with controls has made it clear th at the slowing down and d i diminution of excitability were not due to thyroidectomy. hi the-control group thyroidectomy was immediately followed by a ||tW>g of frequency, an increase of amplitude and a diminution of excitability. Q m fit well into the pattern of the EEG in hypothyroidism, as reported in literature [8]. ' During the 10 days of thyroid substitution these changes did not occur ¡g the experimental animals, except for a slight, not significant slowing down. The changes clearly observable as soon as 24 hours after starting DCPA mitui—•* prove th a t they had been produced exclusively by that treatm ent. Thus, it may be assumed th at the EEG changes noted in the animals jjtb intact thyroid were not due to alterations in thyroid function. The changes in the reticular formation, as determined by EEG, were just g severe ss those in th e cortex. The cortical desynchronisation in response 0G06G22 r ! 5322 72 * U iU tdlT tl]] NO ^ OGübàp-4 5325 5326 0003129 I. TOXICOLOGICAL PROPERTIES OP TORDON* Biochemical Research LabojcafcûEX The Dow Chemical Compan y __ Midland, Michigan C O N T A C T S TfiXAUL , 1963 S E C i & L i ì T O jSX OTHERW ISE CONFIDENTIAL January 15 January ip, i 90j I N F O R M A T I O N SUMMARY AN D CONCLUSIONS t h b D O W CHEMICAL COMPANY 100£ Survival Animal LD„ 50 100£ Fatal g.Ag. g.Ag. g.Ag. Chick (M) 3.98 7.95 approx. 6 Mouse (P) 1.00 7.95 between 2 and 4 Rabbit (Mixed sex) 1.0 3.98 approx. 2 Cavy (P) 2 3.98 approx. 3 Rat (F) — -— 8.2 - range (6.3-10.7) g . A g b ody weight** • DOW 526671 Acute oral toxicity studies were conducted on several species of a n i m a l s . In each case the material was f e d b y single dose' Intuba­ tion as a suspension or slurry In corn oil. A summary of the acute oral toxicity data appears below: ** Calculated by the Well Modification of the Method of Thompson. Undiluted Tordon when applied directly to the conjunctival sac of the laboratory rabbit produced slight to moderate redness of the conjunctiva which subsided between 48 hours and 1 week. The material was essentially nonirritating to the cornea. Tordon was applied to the shaved, abraded and Intact rabbit belly on a repeated, prolonged, confined basis, (continuous contact under a bandage for 11 days to Intact skin and 3 days to abraded skin). Under this conditions of rather drastic exposure, the material produced essentially no skin. Irritation. Tordon was studied for skin absorption properties b y applying It w i t h Dowanol D P M In graduated doses to rabbit skin under an Impervious cuff. Judging b y weight gain and general appearance of the animals, Tordon Is not absorbed In acutely toxic amounts. 5327 # Mamo friT* ^ a M a w m B4 - 0003130 C- 14 SUMMARY OF TOXICOLOGICAL DATA Acute Oral Toxicity Dose 1 fg.Ag.) No. Died No. Fed Capsules 3.98 0/2 Chick Capsules 7.95 3/3 Mouse 20£ suspension In corn oil 1.0 0/2 Mouse 20# suspension In corn oil 2.0 1/2 Animal died ¡2 days after feeding. Mouse 20# suspension in corn oil 3.98 1/2 Animal died within 2 hours after feeding. Mouse 20# suspension In c o m oil 7.95 2/2 Same as above. Rabbit 39.8# slurry in c o m oil 1.0 0/2 Animals appeared n or­ mal during and after fee d i n g . Rabbit 39.8# slurry In c o m oil 2.0 1/2 Animal died 3 days after feeding. Rabbit 3 9 -856 slurry In c o m oil 3.98 2/2 Same as above. Cavy 20# suspension In c o m oil 2.0 0/2 Animals appeared normal during and after feeding. Cavy 20# suspension In c o m oil 3.98 2/2 One animal died overnlte the other three days later* Rat 39.8£ slurry in c o m oil 6.3 2/5 One animal died 4 days after feeding the other 8 days. Rat 39.856 slurry 7.95 1/5 Animal died 3 days after feeding. . 10.0 V5 One animal died 3 days after feeding the other 10 days. 12.6 5/5 Animals okay In one hour but had diarrhea the following day. Animal Preparation Fed Chick In c o m Rat Rat oil 39.856 slurry In c o m • oil 39.856 slurry In corn oil Response-Remarks Animals died overnlte. * Animal was fed 1/2 dose in the morning and 5328 0003131 C- 15 o O Eye Contact - Rabbit Treatment No. of Animals Undiluted Unwashed 3 Very slightly painful. Slight to moderate conjunctival r e d ­ ness subsided in 48 hours to one week. Undiluted Washed with water 3 Washing enhanced recovery in 2 of the 3 animals tested. Response-Remarks 526673 Material Skin Contact - Rabbit Material Condition of Skin No. of APP1 * Site No. of Animals Undiluted Intact 9 Belly 3 No irritation observed Undiluted Abraded 3 Belly 3 Essentially no irri­ tation observed. He a l ­ ing appeared to be normal in all cases. Response-Remarks Skin Contact Absorption - Rabbit (Cuff Technique) Material Duration of Exposure Undiluted 24 hours 1.0 0/2 Material colored skin reddish-brown, otherwise animals appeared normal during and after exposure. Undiluted 24 hours 2.0 0/2 Same as above. Undiluted 24 hours 3.98 0/3 Exposure was made to Intact skin. Response was essen­ tially the same as above. Undiluted 24 hours 3.98 0/3 Abrasions were made the full length of the body 1-2 centlmenters apart. Response was-essentially the same as above. NOTE: Dose No. Died (g./kg.) No. Exposed Response-Remarks Material was weighed out for each rabbit and slurried wi t h a small amount (8-9 cc.)of Dowanol D P M (Dipropylene glycol methyl ether). 5329 0 0 0 3 1 3 ? HI 5330 ■ A !. H E A L T H ' - i .'; / , eiertroItol. Ofic. an.l TreatImrrtici'le In* Health 16 :JJ3, »nine >n Man, 1*151. J. R.: Un indorcs de un do. Bol. Ofic. Nervous System Effects of a Chemical Herbicide mmuniention to P., and Cappel, ratory Animals, Dieldrin Poison:tee on ToxicolH .: Study of ddcs Gilordan, Industr. Hyg. I. DESI. M.D. PROF. J. SOS, M.D. J. OLASZ, M.D. F. SULE AND V. MARKUS C : Estudio de ' dieldrin y in en humanos, . 1957. femorandum on h Service, U.S. n and Welfare, BUDAPEST, HUNGARY T h o u san d s o f new com pounds a re being encou n tered in m an ’s env iro nm ent ail over th e w orld. T h e biological effects o f changes in the chemical en v iro n m en t a re beginning to appear. N ew ly synthesized substances are cau sin g different pathological processes. I t is necessary, th e re fo re , to become acquainted w ith the d am aging effects o f new su b ­ stances on the hu m an org an ism in o rd e r to devise protection ag ain st them . T h e chem ical herbicides a re one o f a group o f the increasingly used co m p o u n d s.' A m ong them , 2,4-dichIorophcnoxy acetic acid (h e re a fte r designated 2,4 -D ) plays an im ­ p o rta n t rple. T h is com pound w as synthe­ sized* in 1944 by Z im m erm an, and its Submitted for |Hil»lirutiaii June 20, 1%1. From the Institute of Pathophysiology (Direc­ to r: Prof. J. So*) of the University Medical Sellout, Budapest, Hungary. ■ i**v 95 utilization as a chemical herbicide was pro­ posed by M itchell and M a rth .' In H ungary it is called D ikonirt and has been marie use o f to an increasing degree since 1952.1 Its molecule is sim ilar to th at o f tryosinc. It ex erts an effect sim ilar to plant horm one. It causes disproportionate grow th and quick death o f certain plants. I t has a selective effect on different plant species. A m ong the m onocotylcdonous dom esticated plants, 2.4D kills the dicotyledonous w eeds.1-1* In 1946 H ild e b ra n d 8 w as o f the opinion that peroral 2,4-D had no pathogenic effect on eith er experim ental anim als o r m an. H ow ever, not m uch later B u c h e r 4 and H ill T found m yotonia, m o to r d iso rd ers, paralyses in th e extrem ities, and gastrointestinal sym p­ tom s such a s vom iting a n d d iarrh ea in experim ental anim als treated w ith this com­ pound. O th e rs observed leukopenia ,3 atax ia an d c o m a 10 a fte r experim ental in trap eritoneal ad m inistration o f large doses o f 2.4D. M ost recently G o ld stein 6 reported on 3 patients in whom grave sen so ry an d m otor sym ptom s, paresthesias, p araly sis, and se­ vere pain ap p eared due to the percutaneous effect o f 2,4-D . R ecovery w as only partial even a fte r years. In H u n g a ry , B o rd a s 1-3 dealt w ith the problem . H e found w eight loss and sym p­ tom s like those m entioned above in his anim als b u t could n o t dem onstrate any ob­ jective changes in w ork ers -spraying 2,4 D. " Sos et al . n *12 have been exam ining the effect o f 2,4-D on thyroid function using radioiodine and increased activity ha> been detected. H ow ever, none o f these studies has dealt w ith the problem o f w hether o r n o t 2,4-D has a dam aging effect on the function o f the central n ervous system . T h e presen t e x ­ perim ents u n d ertook to stu d y this question. ' • 0 •. * • D O W 1511217 A.: The Ef•irc of Rat« to Health 18:263, - •~l . •• < •M M e th o d s Thirty-two white male rajs weighing about 240-250 gui., 5 cats, and 2 dogs were used in these experiments. The electrical activity of the central nervous system was examined hy the use of electroencephalograms in acute and chronic 101 5331 0007167 I I % J I ,• I I •■ 4 » % *; > -i< ». i.-» r.t I 9« H E R VOUS A R C H IV E S OF E S V IR O S M E S T A L H E A L T H c\pi**iife<. Twn frontal ami 2 occipital chvtrodc« were pl.ieol »«I the terehral cortex, ami a hii>olar *id>cnrtic:il electrode wa* placed on the reticular format ion. In the acute experiment* the aninialwcrc auc'thcti/cd with 40 mg. <>l pcntuharhilal (Nembutal) per kilogram of holy weight. ami the elcctrolo were placed on the cortex ami into the MiU'ortex after removal of calvaria. In the chronic experiment* the electrode* were lixed to the top of the skull with dental cement and acrilate, and the electroencephalogram* were taken on unancMhrtizcd animals moving without limitatioit. The ‘pontaneous electrical activity of the animal's hrain as well as response measured in cerebral electric activity to sound and electrical stimuli were recorded. A whistle lasting for 5 sec. was used as the sound stimulus. For the electrical stimulation a s|uare-wave impulse generator was used. The parameters of the square waves were: voltage 1 volt, frequency 100 per second, duration of im­ pulse 8 msec. The entire duration of the stimu­ lation was 5 sec. To examine higher nervous activity, conditionedanxiety-reflex experiments were performed with the method elaborated in our Institute.11 Electric shock was applied as the unconditioned and light a* the conditioned stimulus. 2,4-D furnished by r.ord.i- wax administered in its pure form. In the acute experiment* a dose of 200 mg. per kilo­ gram of l>ody weight was administered intraperitou<".dly on a single wcasion. The same amount was given daily in the chronic experiments until the death of the animal*. In the experiments in which cortical application wa* used, some crystalline granule* of the drug were placed upon the exposed cerebral cortex. Tyro-ine was used for the con­ trols. The brain and spinal marrow of the animals were examined histologically at the end of the experiments. cn f Î /W e r . --------------- — E x p e rim e n ta l R e s u lts I t is well know n that if a norm al anim al ^ u n d er com plete rest undergoes stim ulation, q q the frequency o f cerebral electric activity will im m ediately rise and the am plitude decrease w ith resum ption o f the original activity som e tim e a fte r cessation o f the stim u­ lation. T h is phenom enon is the so-called “ desynchronization.” A ny chemically active agent w hich induces the inhibition o f cere* /v bral functi o f n erv e • desynchror d u ra tio n o abolishes i sen sory ini n eu ro n s ac creases the also a stat o th e r stim I n th e fi? effect o f ra ts w a s < ! •' I H* I ; i ' »» 0»t*ichrmmm- ’’ * ) B » ij /^vftSA Fig. 1.—EEG curve of sleeping ra t Sound stimulus was applied at arrow. A , prior to treat­ ment. B, thirty min. after injecting 200 mg. per kilogram of body weight of 2,4-D; no desynchronization. C, after 60 minutes; desynchronization appears due to sound stimulus. The drawing of the head shows the location and connection of each electrode. The numbers shown are the number of electrodes. 102 VoL 4, Jan., m 2 ’l / Î > ■ Fig. 3.—I (left) and t on the cortf electrodes 1electrodcs 1on the sub: afterwards: decrease in electrode* 1 change wlia; utes; norm.-, sumed; form formation. c 5332 ' V f Disi rl al. f . 1 ll.TU SERYO U S SYSTEM EFFECTS OF HERBICIDE. 97 ¿M» nfr yw<■1« ri- « Fiir. 2 .—Effect of electric stimulation on the duration of desynchronization. The abscissa shows time elapsed since the injection of 2,4-D, expressed in minutes. The ordinate shows the duration of desynchronization in sec­ onds. -pc UUW 1511219 .S' • •• r . •M 'P < Acute .I Sound • treat* 0 mg. D; no -; demuliis. m and imbers I Fitr. 3.—EEC effect of 2.4-D (left) anil tyrosine (right) placet! on the cortex: 2,4-D between the electrodes 1-4; tyrosine between the Hcctrodcs 1 -2 . //, prior to placing on the substances. B, 25 minutes afterwards; with 2,4-D a marked decrease in amplitude between the electrodes 1-4; with tyrosine no change whatever. C, after 40 min­ utes; normal electric activity re­ sumed ; form. rrt. indicates reticular formation. . . I. I t 1 ''>fj Disi ft at. It — — nr 5333 0007169 •'Si. $ / 1• I l I. • I». .\ 98 ARCHIVES OE ENVIRONMENTAL HEALTH i ' • I l jt ■ I 7 :i(ii>n a fte r electrical >limulus liecame sh o rte r, an d , on the .iserage, at the tw entyfilth m in u te a f te r in jection, dcsynchrouization w as abolished. A fte r the lapse o f 50 m in. the resi*onsc could he elicited again. and a t the seventieth m inute it reached the d u r a ­ tion ch aracteristic o f the period p rio r to injectio n o f the d ru g . In the n ex t series o f acute experim ents p e rfo rm e d on S ra ts an d 1 cat, some g ra n ­ ules o f crystalline 2,4-D and, in the controls, cry stallin e g ran u les o f ty ro sin e w ere placed d ire c tly on the cortex. •: • .1 *♦ •• .1 ir> i• i .i Yv In response to 2,4-D the E E G curves 10-15 m in. a f te r application o f the d ru g show ed a m arked d ecrease in am plitude and an increase in frequency. T h e change dem ­ o n stra te d itself only locally between the 2 electrodes betw een w hich th e 2,4-D had been placed. A t o th e r sites of the cerebral c o rte x a n d in the re tic u la r form ation there w as n o change in electrical activity. T he am p litu d e resum ed its original size a fte r 33 m in. on th e average. T y ro sin e , applied in th e co n tro ls, induced no change (F ig . 3 ). In the next exp n ¡m en tal scries chronic expei imeiiis w eie pel formed on S rats, 4 eats, and 2 dogs. D u rin g the chronic experim ents the fre ­ quency o f spontaneous electric activity showed a steady slowing. Instead of the waves having an average frequency o f 7 per second as p rio r to treatm ent, there w ere large slow ...ives o f 2 -per-secund frequency on the tilth day a fte r injection. T h e quick w aves ap p earin g with desynchronization slowed dow n from 12.5 to 4.5 p e r second by the fourth day. A lso the duration of desynchronization w as m oderated day a fte r day. l ’rio r to treatm ent it averaged 32 sec. A fte r the lapse o f 24 hr. the duration w as only 21.2 sec., a 34.2% decrease. O n the second day o f treatm ent the du ratio n o f desynchronization was 14 sec., a 54.5% d e­ crease. O n the th ird day the d u ratio n o f desynchronization w as 6 sec., an 81.2% d e­ crease. O n the fourth day this d u ratio n w as only 4.1 sec., a 91.4% decrease. O n the fifth day th ere was no desynchronization at all in response to the sound stim ulus (F ig s. 4 an d 5) . NERVOUS Si r •t r. 1 Bwf *i.*i' w ; ;c {+-**,**' i ! *1t } M t ■•*> Ijf* I »V vi \ • i L - 4 L - - 'CHRONIC CAI B. ; » »I OX to o ■J to ai *»■ >r; a » ; > 0 « * Ooyt Fig. 4.—A, slowing of spontaneous and desynchronization iref|uciicy in chronic experiment; abscissa is number of days; ordinata is fmiticncy per second. II, decrease in the duration of desynchronization. On tiic fifth day no doynchroui/.atMiii is shown at all. Abscissa shows num­ ber of days. Ordinata shows duration of doynchrom/.atiuii in seconds. Arrow marks onset of treatment. ^ 101 Voi. 4. Jan., 1062 2 Fig. 6.—; O tti et al. 0007170 5334 I 7 1 99 ST.h’t 'V U S S V S T I ' . M l . l / l . l l s Of- l l l . HI UCI P f i I , ,• < I c ÉWw'Vûf. ' 4 * .5 A —* ,Vl,M l ^ w/- •'•ivy. itlfW; *l\ ,. v Vi„-'1^r wjU’''v'*/ *— ’- w ^ - j V ' , , v " '/^W _~''u—' v ^ ? A ^ v j ' . - r . v V / ;"■/ J \ I Z jV'i j*tf- .t to sound stimulus, prior to treat­ ment; duration: 32 see. The entire duration cannot be marked on the diagram. Frequency is 12.5 i>cr second. C, spontaneous activity on the fourth day of treatment; large, slow waves of 2 -pcr-«ccnnd fre­ quency. D, desynchronization due to sound stimulus on the fourth day of treatment, significant shortening in duration. Frequency is 6 per second. Double arrow marks end of desynchronization. 0■ • r ' 7 •■ [ h^/'i I * > * — i"',r ft I — i■■•___ _— i— I— I- • I — I— I I \ I ■ ^ . . » . » CONDITIONED ■ UUW 1511221 1V.J/ ! ^/'l j-1 l; 5 - I:.I 2 .4 - D c b ro u g h t a t quency an in th e cou cells. T y ro chem ically, ever. T h u i specific. Si: w hen place v ery likely and n o t sor precipitatec the d ru g ’s 2.4- D a ex e rte d an acute E E G ished th e r co rtex an d this blockii m in. a f te r m in., it ma; due to 2 ,4-i stream , par with a d c 2,4-D , non: E ven on onset o f t h n f t i et aL r it. jflisILTIi 9 • S r .K l’OUS SVSTF.M EFFECTS OF ULRMCtUH 101 r if • ra ts w as 23% in th e cour>e o f the e x p e ri­ m ent. T h e anim als generally died on the six th day. H istology failed to reveal changes in the g rey o r w hite m a tte r o f the brain o f the anim als. A m yelopathy m anifested as dem yeiinizatiou w as detectable in the dorsal portion o f the spinal cord. W ith Spielm cy cr’s stain in g m ethod the nerve libers showed dem yelinization partly in the pyram ­ idal tract an d p a rtly in the Goll an d I’.urdach bundles (F ig . 8 ). in frqueney and a m arked decrease in the duration o f desynchronization in the chronic E E G experim ents. j!y the fo u rth day the characteristic large, slow, toxic waves ap|>varcd on the electroencephalogram . O n the fifth day no desynchronization m anifested itself due to the constantly increasing toxi­ cosis. O n the basis o f all these finding-, it is very' likely that 2,4-D accum ulates in the organism and in the cerebral tissue and causes a constantly m ore severe paralysis in the function o f the latter. fifth day , no m id be elicited tp c rim e n ts had liac o u tp u t. It d a y an d 35.5 % in g them selves sc m entioned tio n s, how ever, o r fo u rth day ight loss o f the st-r: v £ v rJ O - t v •i .* • ' «. s ' yV,*' In evaluating th ese ex p erim ents it is necessary to sep arate the E E G changes w hen 2,4-D w as adm inistered in trap eritoneally from those o ccu rrin g a fte r direct cortical application. T h e la tte r series o f I t w as im possible to p erfo rm conditionedex p erim en ts w ere p e rfo rm e d in o rd e r to reflex an d E E G studies in one and the same ascertain w h eth er 2,4-D itself o r som e o f t anim al. H ow ever, the resu lts obtained in the m etabolized p ro d u cts form ed in the the 2 g ro u p s o f anim als are parallel. A fte r organism caused th e E E G changes. 24 hr. th ere w as a 34.2% decrease in the 2 .4 - D crystals placed on the cortex du ratio n o f desynchronization and a 2 1 % b rought about a reversible increase in fre ­ decrease in th e conditioned-reflex responses. quency and decrease in am plitude obvious O n the fifth day there is no desynchroniza­ in th e course o f excitatio n o f the cortical tion in response to stim ulation, and the con­ cells. T y ro sin e, w hich is v ery close to 2,4-D ditioned reflexes cease completely. chem ically, d id not cause any change w h at­ T h e question o f the point o f attack m ay ever. T h u s the actio n m ust be considered now be raised. I t is know n th a t the intactness specific. Since 2,4-D also produced changes w hen placed d ire c tly on the cortex, it is o f n o t only the cortex b u t also o f the reticular form ation in the m esencephalon is very likely th at th e 2,4-D molecule itself, necessary f o r both norm al desynchroniza­ and not some o f its decom position products, tion and conditioned-reflex responses. precipitated th e pathological sym ptom s a fte r E lectroencephalogram s show ed the same the d ru g 's in tro d u ctio n into the organism . 2 .4 - D ad m in istered i n t r a p e r i t o n c a l l y changes both in the co rtex a n d in the retic­ ex erted an effect o f a d ifferen t type. In the u lar form ation. S ep arate changes re fe rrin g acute E E G e x p erim en ts it reversibly abol­ only' to lesions o f the cerebral co rtex w ere ished the norm al d esynchronization o f the not dem onstrable. D esynchronization and co rtex .and o f th e re tic u la r form ation. Since conditioned-reflex responses decreased and this blocking began o n the average o f 1 0 disappeared sim ultaneously. T h ese data p e r­ m in. a fte r the in jectio n and w as o v er in 60 mit the assum ption that intraperitoneal a d ­ m in., it m ay 1« assu m ed that the effect was m inistration o f 2,4-D prim arily dam ages due to 2,4-D on cereb ral cells via the blood the reticu lar form ation. T h en , the lesions stream , p araly zin g norm al function. E ater, induced in this region paralyze the function w ith a decrease in th e concentration o f o f the cerebral cortex. 2,4-D , norm al function w as restored. E v en o n the first a n d second day u tte r onset o f the tre a tm e n t there w as a slow ing * f Cl .■ . io •C P F rp m the conditioned-reflex exam inations it is obvious th at 2,4-D also dam ages the higher n ervous activity. T h is effect takes place as early as 24 hr. a f te r ex p osure and by 48 hr. is ra th e r pronounced. B y the fifth day all the conditioned reflexes cease. Comment ttyt I r 'i » • H istology failed to dem onstrate any changes e ith er in the cortex o r in the sub- I'ot. 4. Jan., 1962 Of si ft at. 107 \ 5337 0007173 * 102 •i i» •iV K • I t : e m i u o x m e x t a l h e a l t h v u tlk a l regions, allow ing the conclusion th.it T h e change- found in anim al cxiierim ents n o obvious m orphological change is p ro ­ srtggc -1 the m vd <»f caution in the u-c of 2.4- 1). luciva-cd protection and s o c ia l duced in the n erv e cells by 2,4-1). O bviou-ly n m ro lo g iral exam ination of w orkm en in th e m etabolism an d biological functions of cufilact with 2,4 1) is necessary. th e nerve cells a re dam aged by 2.4-D. T h e possibility o f subm ieroscopic changes in Prof. I. S'"—. In-limit' <>f Pathophysiology, Uni1 the stru c tu re cannot be excluded. ver.-ily M*;ili«.il St I11..I l’.tul.t|t.'-li lliigycs E.-u 9, P.ihI;i| k - i IN. 11lint;.try. T h e histological changes m anifesting them selves in th e region o f the spinal cord R E FER EN C ES m ay l>c responsible fo r the paralyses o f the extrem ities. 1. Honl.t-, S .: F.xamin.itioii of the Occupational ; O n the b asis o f the d ata obtained in Toxic Effect of Plant Protective Agents in Hun- .4 anim al ex p erim en ts it m ust be assum ed th at gary, Dissertation, P.mla|>cst, I960. 2. Rordas, S .: Szcrdahelyi. J., and Sziza, M.: hum an su b jects exposed to th e action o f Occupational Toxic Significance of Recent Plant 2 .4 D also m ay suffer via sm aller dose, Protective Agents in Hungary" I. Experimental m ild er disturb an ces in function o f the nerv­ Toxicologic Studies, Munkavcddcni, 4:14, 1958. J. Bortlas, S .; Kanyo, B.; Nagy, M.t and Weber, o u s system an d o f h igher nervous activity T .: II. Workshop Exposure Studies, Munka- , respectively. vedelem, 4:19, 1958. Functional changes in the nervous system 4. Bucher, N. L. R.: Effect of 2,4-Dicldorodem o n strated them selves a fte r 24 hr., o th er phcnoxyacctic Acid on Experimental Animals, pathologic sym ptom s becom ing obvious only Proc. Soc. Exp. Biol. Med. 63:204, 1946. 5. Drill, V. A , and Hiratzka, T .: Toxicity of on the th ird o r fo u rth experim ental day. 2.4- Diclilorophcnoxyacctic Acid and 2,4.5-TriI t is possible th a t, in laborers w orking with chlorophcnoxyacctic Acid, A.M.A. Arch. Industr. 2 .4 D in w hom rou tin e m ethods fail to Hvg. 7 :61. 1953. d em o n strate pathologic sym ptom s, E E G 6 . Goldstein, N. P .; Jones, P. H., and Brown, studies m ay call atten tio n to early changes. J. R .: Peripheral Neuropathy after Exposure to I t is d esirable th e re fo re th a t w orkm en en ­ an Ester of Dichlorophenoxyacetic Acid, J.A.M.A. 171:1306. 1959. gaged in sp ray in g 2,4-D be protected from 7. Hill, E. V., and Carlisle, H .: Toxicity of ex p o su re to th e poisonous d ru g an d th at Dichlorophcnoxyneetie Acid, for Experimental increased atten tio n be paid to the detection 2.4Animals, J. Industr. Hyg. Toxicol. 29:85, 1947. j o f early signs o f toxicity. 8 . Hildebrand, E.M .: War on Weeds, Science 103:465, 1946. S u m m a ry 9. Marth. P. C., and Mitchell, J. W .: 2.4A fte r th e p aren teral adm in istration of Dichlorophenoxyacctic Arid as a Differential Her2 .4 - D a reversible inhibition o f cerebral hiciilc. Birth. Gaz. 106:224. 1944. 10. Sollmanii. T. H.: Manual of Pharmacology electrical activity w as observed in th e acute ex p erim en ts, a n d in chronic experim ents and its Applications to Therapeutics and Toxicol­ ogy, 8 th Ed.. Philadelphia, W. B. Saunders Com- j th e sam e w as p resen t to a gradually in ­ panv, 1957. creasin g degree. T o x ic E E G signs w ere 11. S6 s. J„ ami Kcrtai, P .: Effect of Dichlodeveloping. A cco rd in g to conditioned-reflex rophcnoxyacctic Acid upon the I 131 Uptake of ex p e rim e n ts, th e h ig h er nervous activ ity the Thyroid, Acta Physiol. Acad. Sci. Hungary 14:367,’l95R. suffered severe dam age. T h e changes are 12. Siis, J.; De-i, l.: Kcrtai, P.. ami Olasz, J .: » p robably produced by th e 2,4-D molecule The Neuroendocrine Effects of Tyro-inc-Analogue itself an d n o t b y som e o f its degeneration Coni|xxinds in Auiiu:il Exvprinicnts, Medical Con­ pro d u cts. T h e point o f attack seem s to be gress, Sopron, 1960, Lecture, unpiihlishcd (Lata. th e re tic u la r form atio n . T h e changes m ani­ 13. Weis*. K.; Vaietiszky, Ss and Weisz, P.: fest them selves as early as 24 h r. a fte r Method of Estalilishinciit of Conditioned Anxiety ex p o su re. Keflex in Rat, Kiserl. Orvosttul. 5:1, 1953. DOW 1511224 ■i .iK cniri-.s o r * 10S 5338 0007174 1 Pollut 5339 f~ -- ■1 DOW 509619 , J on C.3 ----------------- 5340 •- l -CJ o I R * t » * i * t ( t m m t h * J O U R N A L a t th m A m e r U a n V o t f r in o r y Alecflcal A s to t/v ffo n , V ol. 1 4 3 , N m . 4 , A vg. 15. 1945, p p . 39S »399a DOW 509620 Chronic Toxicity cf 2,4-D Aiksnclamine Saits to Cattle J. .S’. Palmer, D.V.M., M.P.H. A vaii^uile uata ind icate th e low toxicity of ch lo rin ated phcnoxvacctic acid com­ pounds to c a ttle .’ *'® These conclusions are based on re su lts derived from d irect oral dosage o f c a ttle o r from exposing cattle to p a stu re s sp rayed w ith v arious hcrbicidal co n cen tratio n s. T he hazard is negligible when h erb icid es a rc applied in concentra­ tions th a t effectively control dicotyledonous plants. M any different, hcrbicidal compounds have been introduced on the m arket, but 2,4-D w as th e first of its general type to be accepted. I t h a s been used in v ast am ounts fo r co n trolling annual weeds in cereal crops an d p a stu re s and also fo r weeds and b ru sh in t u r f and ranges. Re­ search d ata on o th e r anim al species indi­ cate th a t it is relatively m ore toxic than o th e r com pounds . 1 T he likelihood o f cattle g ain in g access to in to x ic a tin g am ounts of th is herbicide in sprayed areas, sp ray tan k s, and storage co n tain ers prom pted th is study. regimen, was calculated at a rale oi 50, 100. 200, ami 250 ing./k". nf body weight. Tlic daily dose oi llir liijuid herbicide varied from 12.5 to 70.5 Cm. and was arlinini.-tcrcd with a do-c syringe, using water as the vehicle. Each steer was restrained in a squeeze chute to assme accurate dosing. One steer each was treated with 50, 100, and 200 mg./kg. and 2 steers with 250 mg./kg. An untreated steer was maintained as a control. Results One s te e r tre a te d in itially w ith 70.46 Gm. (250 m g ./k g .) w as elim inated from the study. A chronic re sp ira to ry infection developed, considered as in cu b atin g before th e te s t w ith th e focal p o int in th e pharyngeal region. T he herbicide tre a tm e n t probably acted as an ir r ita tin g fa c to r and se t up a fu lm in atin g condition. The c h a r­ a c te ristic signs of labored re sp ira tio n and w eight loss w ere observed. The ste e r was tre a te d a f te r 1 weeks w ith ap p ro p riate th e ra p y and then sold a f te r re g a in in g w eight. Also, pneum onia unexpectedly de­ veloped in the control steer, b u t an tib io tic Materials and Methods th e ra p y brought quick recovery. OÎ 6 yrnrlinx 5trrrs uriphins 300 to 10il 11>. u*rtj A direct relatio n sh ip w as noticed be­ initially. 5 won* IIoUtcin*Fii(*'ian>, anti ] was a tw een th e q u a n tity o f daily intake, the ac­ IlcioOml oivMt. The stooi* wort* foil a mnint«‘ti;uirt.< cum ulated total, and th e length of the ration t of a prolfiti roncontiatc r:»ili inmiiin" amt a comlmiuluMi of Uo;-;ui anti alfalfa hay oat h treatm en t period. W ith Ihc increase in daily dose, less tim e w as required to pro­ eioitiii". Tlu: talion wn* h )mii weight Jnu was itoiortnl ill liio |»t in*'i|>7tU a* well a* in |l«t rnn* duce signs of poisotjing so that the total am ount ingested was decreased (T able 1). Irnl. WVi^htft won® «Irtoiinilird wroklv. 'I ho 1Î.M) t»M ht't hiritit roiilaiiiO(| (tV e alkatiola T he s te e r treated w ith 50 mg. I;g. was mill«* sail.* Tin* daily «!«»*<•. on a given 112 daily tre a tm e n ts (5 per week) ra n g in g from 12.5 to lit.7 (,’m. Ih irin g l?rr»w Tot!io>]<>!,.ir<«l I»>\< Aniin .i l D i - t i i v th is lim e, the s le e t's w eight increased Aft«l K r M f f ! i D iv is io n , A;:« i c u l t m *1 KYm -h h I i S ( tv iu > , U S D A , K r i m i l c , T r t . r . . from to fiti:'. 111. The treatm en t period Tin* %tfiler t h . t n k s lit ■ l- b o if tr- V v r m p î« *y r ts W . Rec* extended from Ju n e 25 to l>i-c. ■!. 1062, t u f , T. S f t o t , anil I t. J o i n s in m ii i. t ii n .: ti»«*s Mmly. •Tl»v | r < t htflnridf was a rtiinm m t.il prt.;lm l. w ith an approxim ate averag e of 4.5 treatla lu ll t» ! W r r i l KiMrr, 40,'* p r i n t m ' i d men Is weekly. When the stu d y ended, this b y lit.- l)*iw C l t c m i r a l C<«., M h I1.i «u !. M i d i 398 J.A.V.M.A., Vol. 143, No. 4 5341 000287S I c TACLE I— Chronic Toxicity of 2,4-D AH'onolominc Soft to Cottle-—*Rclationihip of Dcily Oor-c to Ac­ cumulated Doicgc A ccum u lated Daily dose dolale (Gm.) To produce intoxi* Total cation 1*29.3 261C.0 Unaffected Poisoned 70.5 1958.6 1049.1 Disqualified Poisoned Poisoned lìasi«. (niR./lig.) Variation (Gm.) 212 SO 100 12.5-19.7 25.9*35.7 « 112 IS 2546.4 290 200 250 70.3 54.2*58.7 51.8*53.3 34 15 i 44 20 1419.fi 714.3 213 2U 21) 21« 21!/ rnvm nr __ il * •Given daily 5 days a s te e r hud no appnront signs o f intoxication. C attle given doses of 100, 200, and 250 m g ./k g . h ad sig n s of poisoning a f te r 8 G, 3-J, an d 15 tre a tm e n t days, respectively. T he m ore heavily dosed steers w ere tre a te d to te rm in a tio n of th e experim ent. T he s te e r given 100 m g ./k g . had to be m edicated a f te r it developed a digestive d iso rd e r ch aracterized by extrem e tym ­ pany. Tw o days w ere req u ired to correct th e condition, although anorexia w as never observed. R um en atony fro m accu­ m ulated herb icid e w as ap p aren tly th e p re ­ disposing fa c to r; th e ste e r w as removed from th e exp erim en t and m ade an uncom ­ plicated recovery. P rim a ry sig n s of intoxication in steers given 200 and 250 m g ./k g . o f th e com­ pound w ere developm ent o f d ry cracked m uzzles and ulcerated mucous m em branes, w ith ep ista x is on ro u tin e re s tra in t. A clear, viscid nasal discharge, associated w ith a fe tid odor, w as noticed. P ro g ressiv e a p a th y an d depression w ere ap p aren t. One s te e r given 250 m g ./k g . had m oder­ a te ty m p an y associated w ith m clena. N e­ cropsy revealed severe hem orrhages in the larg e in te stin e and pulm onary congestion. T he s te e r given 200 m g ./k g . had m uscular w eakness in th e h in d iju arters and a sta g ­ g e rin g g a it, d ro s s pathologic changes noticed at necropsy w ere inflam m atory sw elling o f th e kidneys and a pneum onic condition considered to be the im m ediate cause of d eath. Sum m ary a n d Conclusion« L D aily oral doses of all.anokim ine salts of 2 , 1 -dichlorophorioxyacrtie acid ( 2 , 1 -D > w ere given to 5 yearling stecra to de­ te rm in e th e chronic pixiciiy of |h is eomAugust 15, 1963 Results pound. T he doses w ere based on m illigram s o f 2,4-D p e r kilogram of body w eig h t and w ere ad m in istered by dose sy rin g e, w ith w a te r as th e vehicle. Doses w ore given daily 5 days a week. S igns o f poisoning w ere noticed in the m ore heavily dosed steers (250 m g ./k g .) a f te r 15 tre a tm e n ts, in c o n tra st to 86 tr e a t­ m ents at 100 m g ./k g . A dose of 50 m g ./k g . h ad no a p p a re n t effect on 1 ste e r a f te r 1 1 2 doses. A d irect relatio n sh ip w as th u s de­ tected betw een th e relativ e q u a n tity of daily in ta k e to th e accum ulated to tal dosage and th e len g th o f th e tre a tm e n t period. I t w as concluded th a t th is h erbicide h as a low degree o f chronic toxicity, a p p a re n t­ ly because i t is n o t read ily stored in bovine tissu es. However, i t is possible fo r cattle to ingpst enough to produce m o rb id ity and m o rtality . DOW 509621 Sucr Nu. No. treatment** To produce imoxi* Total carimi R a fa ra n co i •Rock. W. H., Itiniis, W., .lame«. T_, and William?. M. C : Rc*nlts nf » ed ili" lleiliicidc-Ticalcd Pianti­ lo Calie? and Slut*|i. J.A.Y.M.A., 1311, (March 15, ] % 1 1 : 320.3ÌI. 5Crafls. A. .S.: The Chcmisliv ami Mode of Action nf 1(•-■'Incide*. Iiilcr-rimrc l'nhlislirrc, New York, N. Y.. ]%1. i(»ritf*«liy..Tì. li., and » liv id i, K. 0.: Suine EfTcct? nf llriliii idi-- mi Pa-litri* ami un dra/iii" I.ivestoek. Micidiali Agi ir. Kxprr. Sia. Qnail. Unii. •'■2. (19301: .".:i:-:i!:r>. mnd (’•articii-, *Mii. dirli. J. \T„ ]Inil^'.«*ti, R. Tolerauto of Kami AniniaU I"• Kred ContainC. ing 2.1 •1lit Moi ii| I n . i f riti* Arid. J. \ iiíin. Sci.. 5. (Pilii.l : Z ‘liad. •i.ir. I?. 1).: \«Uanri-s in Veit*i ¡iin ly Srirnrc, Vid. I. 11..- To>icily of Ili-cv lieidi -■ an- 1 j lerlnritlc. lo hive*■11» k., Arailoniu: Pic-a Ine., NV\\ Y.nk, N.Y.. fl9.'i.,;i : 2('> m*(*• ‘Iti.n .*, v . K.. ami ÍT\itiu*>. T. A. Summnrv of Toxirul o/irul iiifurinrfion on 2 ,b l) ami 2. ;.r.-T Typo ilrilthidr* ;alni an alii.iti<•11 of the limititi? to 1.¡w -tt••.1. A»•o'iat*-i| uitli *! !lift! V* Ain. J. Vet. i b - ., i'., ( O ri., io :,!) ; i. 399 0002879 U i, Jf »«i í 4 I ot p Vjj ii --- -V; ro/i; Toxicity of 2,4Dichlorophenoxyacetic Acid for Man and Dog O JOHN K. SKABIKV. MO tí dee ' hit: O ■NJ O NEW ORLEANS ^>5 ID S that ;>or: the cinc tion ¡'fin agre that 'he prep. was .n'et: mgr li­ 3 H a 3 000630! ir» •H ?■ » 4 In N ovem ber, 1959, Goidstein. Jones. and B row n *1 reported three cases o t peripheral neuropathy follow ing exposure to an ester o f dichlorophenoxyacetic a d d (2 ,4 -D ). Tlie first patient had spilled 60 ml of a 1 0 C^ solu­ tion of an ester o f 2,4-D on his forearm s w ithout w ashing it off. T h e patient felt unduly fatigued on the evening of the same dav, and w ithin the n ext ten davs he developed nausea, yom iting, and 2 0 -pound weight loss. A pproxim ately two months later he had a sim ilar e x j» su re on his legs, ami he developed nausea, vom iting, and diarrh ea d u rin g the subsequent five days. O ne week a fte r the second exposure, he com­ plained of num bness and aching of the digits of all extrem ities. T h ere was desquam ation of the skin of the palm.- and soles, and within six weeks a fte r the second exj>o>ure he had well-developed neuropathy. *1 he second patient had a less determ inable am ount of exp.osure by derm al 'vetting with kui'm it:«! for ¡-uhlic.atir.t; Aug 10, 1002, i m res-n r of Medicine. P q'-.irtnent of M rlicute. I.i.nlji.ir.a Slate t.'iiivcr-uy School of Medicine. an ester o f 2.4-D. Shortly a fte r exposure, she noted swelling and aching o f the feet and legs, which persisted for approxim ately I '/i weeks. A pproxim ately one y ear later she had a sim ilar exposure to the herbicide with recurrence of d inilar sym ptom s. However, she later developed anorexia, loss of 20 pounds in weight, and severe pain in her iegs with painful swelling of the m etacarpal joints of both hands. A pproxim ately five months a fte r her second exposure she had a riaccid paraparesis. T he third patieiir reported had been spray­ ing with a solution o f an ester of 2,4-D. D uring this activity his sleeves and trouser legs had been wetted with the herbicide, and some of the spray m ight have been inhaled. O n the following day, he noted malaise, headache, nausea, and vom iting. V ertigo was severe on the secuml day following - praying. Paresthesias in the extrem ities and p.«in m the legs appeared within fo u r or five days, followed by tw itching of the muscles in the caives ami am is. K ascvulations became generalized. T here were m> neurological or to 2 ■.■lair iu e r dichi re norrr XI. : hera no ti­ the t the ; know cocci t'/CUc that cours- were ‘her.'1 .' *.*ni y ñ rs: ^JSi'l. TÍJctic C^i.id ., list, kept pare;o.ch: 8:. J.i n TOXICITY a . ¿1 sf :vr • f-t •• I ■’T ^ J ll 2- r‘ M :n *:i\ ".5>; ‘‘ •V ? * .e • he - ■I .•* t ‘.V .< ** JIU tv -VTjHf• ■*• * ' • » *5 \ 'Ì f p r.' ■/•r's n •t ?» ^? , *V v e x p o su re, M ie f e e t a n d r \- .¡u ia te ly 2 y 4 ^ ¡ it e r » h e h a d , lucide w ith H ow ever, of 20 i h er legs >;sl jo in ts ’ : c m onths *.i-: a Ilaccid : vm Sprayr of 2,4-D . . ;:.'l tro u se r jrr.icide. am i »Tii inhale*!. a-d malaise, * rti-o w as 'I'rt-ying. pain in : . \ e iltlVS, . / in the became .If or » ,i ' ; ? i > electrom yographic changes, hut his sym p­ toms persisted. In a subsequent com m ent 5 on this report of ¡leripheral neuropathy follow ing exjiosure to an ester of 2,4-D , GoWlwater points out that these a re the first such cases to be rel*>rted and that one w ould have to know the com position o f the vehicles, solvents, and emulsifiers used in the com m ercial p rep ara­ tion b efore incrim inating 2,4-D as the principal toxic com pound. G oldstein ct al agreed w ith G oldw ater’s com m ent and stated that in one instance they w ere able to obtain the label fro m the container o f the 2,4-D preparation. O n this label, the preparation u as stated to contain “ 2,4-dichloruphenoxyacetic acid, isopropyl ester 44 c/c and inert ingredients 5 6 % .” Because o f these reported cases of toxicity to 2.4-D , and the recent study of a man claim ing disability o f a different type subse­ quent to sp ray in g w ater hyacinths with a dichlorophenoxyacetic com pound, I decided to rep o rt certain therapeutic trials w ith plant horm ones carried out in 1948 and 1949. M y in terests in plant horm ones w ere therapeutic ra th e r than toxicologic, and since no therapeutic efficacy was dem onstrated in the trials, no attem pt was m ade to publish the results. In 1948-1949, there was no known th erapeutic agent fo r dissem inated coccidioidomycosis, histoplasm osis, o r cryp­ tococcosis. It appeared rem otely possible that the p lan t horm ones m ight alter the course of these fungous infections. Since we were unable to find any evidence o f therapeutic effectiveness in the literature, and only m eager rep o rts o f anim al toxicity, I first screened a m ix tu re of indole-3-acetic acid, indole-3-propionic acid, a-naphthaiene■icetic acid, and 2.4-dichlorophenoxyacetic acid in guinea pigs infected with C nccidloidcs im m itis. N o therapeutic effect was noted. S ix fem ale m ongrel dogs, of ap p ro x i­ m ately one y ear of age. w ere infected with fl¡sto tU ism a caf'sidam m . T h ree dogs w ere kept as controls, and three were treated pnrenterally w ith the sodium salt of 2,4dichlorophenoxyacetic acid. O nly the treated dogs are of interest here. D og 1 received a total of 464 m g o f sodium 2,4-D in 28 injections over 37 days, of which 2S8 ing were given intravenously. O n a weight basis, the injections am ounted to 1.17 m g /k g of body weight. D og 2 w as given 389 mg in 24 injections over 32 days, of which 261 mg were given intravenously. O n a weight basis, this am ounted to 3.2 m g /k g of body weight per injection. D og 3 received 3S9 m g in 24 injections over 32 days, o t which 277 mg w ere given intravenously. T he dose per injection was 2 .6 m g /k g o f body weight. Dog 1 was autopsied 202 days a fte r the first injection o f 2 ,4 -D ; dog 2 was autopsied on the 8 6 th Ja y following the initiation o f treatm en t; and dog 3 was autop-ied on the 85th day. All dogs had gained substantial w eight du rin g the experim ental period. All dogs showed evidence of visceral histoplas­ mosis at autopsy, but th ere w ere no gross o r microscopic lesions in the liver, spleen, lungs, kidneys, o r adrenals other than those due to histoplasm osis. N o neurological tis­ sues w ere exam ined. A m oribund patient w ith dissem inated coccidioidomycosis w as selected for in tra ­ m uscular injection of auxins. T his patient received a total o f 40 m g of the sodium salt of 2,4-1), 3.3 m g o f indole-3-propionic acid, 3.3 mg of indole-3-butyric acid, and 3.3 m g of a-napiuhaleneacetic acid d u ring a period of fo u r days. D eath occurred on the fifth day. T h e patient exhibited no unusual sym p­ tom s d u rin g treatm ent, and the autopsy findings were only those of generalized coc­ cidioidomycosis and hypoxia. DOVI 1392107 ce 203 In A pril, 1949, I decided to treat a 4Syear-old male patient w ith dissem inated coccidioidomycosis w ith auxins to the limit of tolerance. It was obvious that the patient would die w ithin the n ex t several weeks unless the course o f the disease could be modified. T he details o f treatm ent appear in Table 1. Intravenous im'iidon of 2 gm of 2,4-D produced no changes in his physical examination and pro­ voked no complaints from the patient. When 3,600 me of 2 ,4 - 0 were ¡riven intravenously dum p a period of two hours, a reaction appeared. T. .trine the latter part of the infusion, the patient heemre very (piiet, .'emisliit»'roiis, and exmhited fihr.uarj 5345 0G06906 / / 392108 J 204 ARCHIVES OF T a ble \ D ue i : J .3 > 4 Routeof Administration l. — Administration af Auxins to Man 2.4-D, M« IM IM IM IV IM IV IV IV IV 3/ 4 /« 3 / 3/49 3 / #/49 3 / 7/49 3 / 9/49 3/10/49 3/11/43 3/13/49 3/13/49 3/1V49 3/19/49 3/21/49 3/23/49 3/24/49 3/23/49 3/28/49 3/2S/49 3/30/49 4/ 1/49 4/ 2/49 4 / 4/49 4 / 3/49 4 / i/49 rv !'/RO.VH EX T AL HEALTH Indoi*4-Üutyrt« Acid, M ( a e n k Acid, M | I 2« 0.M 0.64 l.M l.M 1« 160 24 120 120 240 440 400 IJ2 c.a 132 l.M l.M \Z 2 13.2 13.2 IV IV IV IV IV IV IV IV IV IV IV IV 2.000 rv ZjOO 6.94 13-2 MO MO soo soo soo 33 33 33 33 3J 33 33 33 33 33 900 SOO yo soo soo soo React** * A— :£■ Local buruJcf rrocsino added—ooo« & ■*' « .V:S Nooa Nooc Non* Nona Nona Nona NJ-D TOXICITY L HEALTH • . -s- Tabi.e 3.—Subacute Toxicity of 2,4-D for ■inimats •¥ j \ Animal to d •- No. Intraven­ ously 3 J 0 X I L D i .. 100 X - LU ¿0 X ®(l died, 1 sjcriflu«d very 111) IS x <1(sacrificed very ill) 2 2 s \.*fr i i llet U'■ t produced in the hum an patient and that proI duced in the two m onkeys. H ow ever, on a w eight basis, the dose adm inistered to our patient intravenously was approxim ately 6 6 m g /k g o f body weight whereas the m axim um intraperitoneal Jose given to the monkeys was 428 m g /k g o f body weight. A lthough the m ajority o f studies o f acute toxicity o f the 2,4-dichlorophenoxyacetic com pounds are in d o se agreem ent, the num ­ ber of anim als tested by some au th o rs was smail, and some disagreem ents ap p ear which cannot be resolved on the basis o f the data presented. F o r exam ple, the acute I .D-,.. for oral adm inistration o f 2,4-D to dogs was 100 m g /k g according to D rill and H ira tz k a s based on the study of fo u r anim als. O th er w o rk e rs ' gave an am ount of the butyl ester of 2.4-D calculated to contain 500 m g of 2.4- D per kilogram to each o f two dogs w ith ­ out observing any deleterious effects w 'thin *>6 hours. Clue of the two dogs was autopsied and showed no evidence of ar.v m acroscopic lesions. T he other animal rem ained in apxcellent health fo r the follow ing 0 0 0 6 9 0 » ' m l> 5348"' . vP rîi - - - y DOW 1392111 ■■ » i n TOXICITY 'E .lt.T H 'h ' _nipt<>ms ce nbs. In „ ru b b in g St ! T h o se :ll ', ; 1.5 grn -• ) te intra- m l •a\ :e ( j I vom itm ptotns t-f gs, with ,• ,ftharjjy, the eyed about tV sred fo r . îtensitv. ■is com* iv.ved 12,V je Jy p rio r . a ; ]{ ra. tare th a t ^ sou ¡urn • -Î .* i n . voiced v.,, lated to ¡c was a reaction .,-h.nt pro­ per. on a : 1 to our ately 66 r .taxi.-mim . : U'lnkeys r of acute , 'Xyacetic ,) : he uum; i" i' was . ■ which ’•he data A fur ” e-' was r.uzka 3 w*; O ther ester mg of . - with' within ,.:i!..p'ied i "'Cupic in aj>, "¡¡"wing ' 1 1I rj ‘. , 1 I / 1 1 I 1 I 1 1 i11 207 s j days. O ne would hesitate to cunciude ft'iuii these data th at the butyl ester is less t..\ic than 2,4-dichlorophenoxyacetic acid. T he lowest dosage adm inistered in trav e­ nously to dogs (25 m g /k g ) over a period of ,-ix days was toxic ,4 producing decubitus ulcers o f the extrem ities and necrotizing le­ sions about the m outh. A low er lim it for -ubacute toxicity in dogs has not been estab­ lished. A ccording to D rill anti H iratzk a * the largest dose o f 2,4-D w hich could be given i>rally to dogs fo r extended periods w as 10 n ig /k g o f body w eight. C om m ercial herbicidal p reparations o f the chlorinated phenoxvacetic acids and associ­ ated com pounds are usually form ulations of ■.oe o r m ore active ingredients with solvents, dispersing an d w etting agents, and some­ times co-solvents. T hese may contain as active in g red ien ts either the acids o r their -alts, esters, o r am ines. T h e exact com posi­ tion o f liquid form ulations is o ften not stated in labeling. A lthough Rowe and H y m a s 9 -tated that the acute oral toxicity o f the commercial form ulations of the D ow C hem i­ cal C om pany tended to be projiortional to '.hut expected from their content of active in­ gredients an d th at the "in e rt in g redients" did not ap p e a r to e x e rt a potentiating effect upon toxicity, H ill and C a rlisle 4 found that the toxicity o f 2,4-dichloronhenoxyncetic acid dissolved in tributylphosphate and oil was much g re a te r than the toxicity o f 2,4-D .done. T h e increase in toxicity was due to the tributyiphosphate-oii m ix tu re itself. T he sim e au th o rs p erform ed sim ilar studies w ith A"-butyl alcohol-oil solutions of 2,4-D , and 'Cited "A lth o u g h not conclusive, these e x ­ perim ents indicated th at X -butyl alcohol-oil concentrations of 2,4-D are at least o f the sam e o rd e r o f to x ic ity ’"as the tributylphosphate-oil solutions." T hese findings by H ill and C arlisle lend strength to G oldw ater’s criticism 3 o f the report by G oldstein . 1 B u ch er ,3 w ith o ut indicating the num ber of anim als studied, stated th at the histopathulogy produced by 2,4-D in acutely in­ toxicated mice consisted o f wide dilatation of the blood vessels of lungs, liver, and kid97 5V-'nii-v neys. ¿ h r noted moderate atrophy of the liver in one chronically treated dog. There was no significant variation in the peripheral blood of chronically treated mice from that observed in controls with respect to hemo­ globin. red ar.d white blood cell concentra­ tions, and differential counts. H ill anti C arlisle 4 found thac rats and guinea pigs dying of massive doses of 2,4-D consistently exhibited congestion o f the vis­ cera and enlarged swollen kidneys. M icro­ scopically, there was m assive cloudy swelling o f the proxim al convoluted tubules w ith cast form ation. T he glom eruli and blood vessels w ere unaffected. Occasional anim als showed slight patchy pulm onary edem a and alveolar hem orrhages. Rabbits that received injec­ tions of one fourth of the L D 50 parenterally fo r from 4 to 13 days showed slight-tom oderate degenerative changes in the p ro x i­ mal convoluted tubules. Scattered petechial hem orrhages were present in the lungs of a few animals. T hree to seven daily injections of one half of the L D ;<, produced a signifi­ cant decrease in hemoglobin in fo u r o f six rabbits. T he renal changes w ere sim ilar to those observed in the rabbits receiving one q u a rte r of an I.D -U fu r a longer period of time. in addition to the developm ent o f bleeding and necrosis of the gum s and decubitus ulcers, dogs also exhibit evidence of liver dam age more frequently than o th er animals studied .3’3 T he histopathology varied from occasional areas of focal necrosis in animals subjected to chronic oral toxicity studies to extensive necrobiosis throughout the liver in one dog 4 given two daily injections of 200 m g /k g of the sodium salt o f 2,4-D . F o r the most part, tings succum bing to massive doses o f 2,4-D showed centrolc.bular degeneration, atrophy anti lysis o f the parenchym al cells about the central veins o f the lobules, and congestion and dilatation o f the paracentral sinusoitls .4 Dogs also showed cloudy swell­ ing of the renal tu bular e p ith e liu m ,° anti one dog receiving a m assive do>e 4 showed actual tubular epithelial necrosis. Lymphoid necrosis was observed in the acutely poisoned o b o u sio 1* i i l: i• i< t: j t i. 1* I, (' >. I I 5349 .¿g S S g g g / * $ § S ¿* * ,vr^*s.v< * ,*•. ***7 ■ •••-i?» ■..T.-...>Tv-'V' ■ ; % '*■■»•-? •V-.'Jr5'5 392112 --08 >• i r ». ARCHIVES OF ENVIRONMENTAL HEALTH.'?'] dogs, an d some o f them showed term inal lym phopenia in the peripheral blood. R eferences to hum an toxicity are meaner. H ild e b ra n d 10 m ade the sim ple statem ent “ Some investigators have consum ed the chemical and suffered no ill effects." Assouly 11 stated that w o rkers employed in the fabrication o f an ester o f 2,4-dichlorophenoxyacetic acid developed sym ptom s of som­ nolence, an o rex ia and gastralgia, increased salivation, a sw eet taste in the m outh, a sensation o f drunkenness, heaviness of the legs, and hyperacusis. W h eth er these sym p­ tom s w ere related to the 2,4-dichlorophenuxyacetic acid o r to the chemicals used in the process o f esterification is not known. C ertainly a sw eet taste in the m outh with sialarrhea suggests the sym ptom s were re­ lated to the chemicals used in esterification. H yperacusis w as not noted in either of the tw o patients reported here who received parenteral 2,4-D . M itchell, H odgson, and G aetiens 12 state in a footnote "D r. E. J. K rau s of the U niversity o f Chicago reported on Jan u ary 26, 1946, that a hum an (adult male") had taken daily by m outh, 5G0 mgm. of purified 2,4-D either a fte r the m idday o r evening meals d u rin g a period of 2 1 days w ithout experiencing ill effects." . .* / S u m m a ry T hree dogs with experim ental histoplas- ' Vmosis w ere screened fo r chronic parenteral toxicity o f sodium 2,4-dichlorophenoxyacetic acid. T he dogs were given parenteral sodium 2,4-D at the rate o f 1.17 m g /k g o f body weight, 3.2 m g /k g o f body weight, and 2.6 m g /k g of body weight per injection. T re a t­ ment extended between 32 ami 37 Jays. In .. y this dosage, there was no evidence of chronic toxicity. O ne patient was given a total of 40 mg of the sodium salt of 2.4-D. 3.3 m g of indole-3propionic acid, 3.3 mg o f indole-3-butyric acid, and 3.3 mg of a-naphthaleneacetic acid intram uscularly during a period o f four days. Xo toxicity was observed. A second patient w as given a total o f 369 mg o f irdoIe-3-butyrie acid, 33.3 mg of «naphthaleneacetic acid, arid 12,712 mg of the sodium salt of 2,4-dichlorophenoxyacetic acid parenteraliy d u ring a period of 34 days w ithout-observable toxicity. M ost o f this m aterial was given intravenously. Tw o days later. 3.6C0 mg o f sodium 2,4-D were given intravenously and produced definite clinical signs o f toxicity. T he p atient recovered clinically’ from this toxic episode within -'S hours, and showed no fu rth e r evidence of neurologic o r m uscular change in the sub­ sequent two weeks of life. D eath was due to dissem inated coccidioidomycosis. The autopsy fimlings are described. A single intravenous dose of 2.000 mg of sodium 2.4-1) did not produce clmical evi- 5350 r m./ • r p! ->. A. 2,4 t 2.4iUs TT69000 It would seem logical that chemical modifi­ cation of the chlorinated phenoxyacetic acids m ight well result in altered pharm acological action and toxicity. A n interesting exam ple o f this is th e dim ethvlam inoethyl ester o f /’-chlorophenoxyacetic acid. T h is ester was produced by T huillier, R um pf, and Thuillier.la T h e ester has an L D ;i, to r mice of 330 m g /k g when given intravenously and an I.D ja o f 845 m g /k g when given intraperitoneally to dogs. Chronic toxicity studies in rats indicated that 300 m g /k g could be given for m ore than five m onths w ithout alteration of the blood cell count o r of the organ system s. H ow ever, it ap|teared to act as a central nervous system stim ulant and was capable o f raising a lowered arterial blood pressure. T h ere was no evidence of a m yotonic action. Coirault, P ournre. Dam asio. R o u it, Deligne. David, and Tnlairach 14 reported using this ester therapeu- tically on the neurosurgical sendee o f two “ hospitals in the m anagem ent of stuporous ■ and comatose patients. T he oral dose for . /,] man averaged 400 to 1,200 m g in 24 hours, btu doses of 4 gm have been given without reaction. T he ester was given intravenously in doses o f 250 to 500 m g at the rate of about ; » 250 mg in five m inutes. T h e authors state v '_. that this material had beer, given for from eight days to six m onths w ithout a single reaction. No inform ation is given concern- v-? ing the possible herbicidal activity of this -V . chlorophenoxyacetic acid ester. ■'J. •EAI.TH. V Ot two .uporous .. lose fo r •'V *, :4 hours, > * ; . w ithout venously of about 'V ■>rs state ror from • a single concern/ . o f this '\Vf 'J'i. . .*■<• :areiiteral ••xyacetic i* il sodium uf botiy •. and 2 .6 n. T reatJays In ■i ch .»ic 49 mg of indole-3-3-butyric ■.ce’.ic acid /•lur days. »i* • ' ... 2C9 : j -d t o x i c i t y 7. Eyzaguirre. C ; Folk, B. P. ; Zierler, K. L.; and Lilienthal, J. L., Jr. : Experimental Myotonia and Repetitive Phenomena: The Veratrinic Effects of 2,4-Diciilorophenoxyacetic Acid in tlie Rat. Amer J Physiol 153:69-77, 1948. 8. Baker, D. L.; Ramsey, F. K.; and Syl wester, E. P. : Suspected Poisoning of Dogs From Eating Grasses Treated With 2.4-D, N Amer Vet 34:194, 1953. John H. Seabury, MD, Department of Medicine, 9. Rowe, V. K.. and Hymas, T. A. : Summary of 5 hool of Medicine, Louisiana State University, Toxicological Information on 2,4-D and 2,4,5-T . - . l Tulane Ave, New Orleans 12, La. Type Herbicides and an Evaluation of the Hazards to Livestock Associated With Their Use, .Amer J Vet Res 15 622-629, 1954. REFERENCES 10. Hildebrand, E. M. : War on Weeds, Science 103:465-468, 1946. 1. Goldstein, N. P .; Jones, P. H .; aud Brown, 11. Assouly, M. : Désherbants sélectifs et sub­ J. R.: Peripheral Neuropathy After Exposure to stances de croissance; Aperçu Technique; Effet an Ester of Dichloroplieroxyacctic Acid, JAMA pathologique sur l'homme au cours de la fabrication 171:1306-1309, 1959. de l’ester eu 2-4D, Arch Mal Prof 12:25-30, 1951. 2. Goldwater, L. J . : Peripheral Neuropathy, 12. Mitchell, J. W. ; Hodgson, R. E ; and GaeîJAMA 173:87, 1960. jens, C. F.: Tolerance of Farm Animais to Feed 3. Bucher, N. L. R.: Effects of 2,4-Dichloro- Containing 2,4-Uichlorcplier.oxyacetic Acid, J Ani­ lihenoxyacetic Acid on Experimental Animals, Proc mal Sci 5 :226-232, 1946. Soc Exp Biol Med 63:204-205, 1946. 13. Thuillier, J.; Rumpf, P. ; and Thuillier, G.: 4. Hill, E. Z„ ar.d Carlisle, H .: Toxicity of [Derivatives of Acid Regulators of “Plant Growth: 2.4Dichlorophenoxyacetic Acid for Experimental I. Pharmacologic Properties of the DimethylaminoAnimals, J Industr Hyg 29 :S5-95, 1947. ethyl Ester of p-Chlorophenoxyacetic Acid (235 5. Drill, V. A., and Hiratzka, T .: Toxicity of ANP) I, C R Soc Biol (Paris) 133:1914-1918, 1959. 2.4OichIorophenoxyaeetic Acid and 2,4,5-Trichlo-14. Coirault, R. ; Pourpre. H. ; Damasio, R. ; rnphenoxyacetic Acid, AMA Arch Industr Hyg Rouif, G. ; Deligne, P. ; David, M.; and Taiairacli, 7:61-67, 1953. J. : [The Treatment of Consciousness Disorders in Neurosurgery by the Dimethylamincethyl F.ster of 6 . Bjorn, M. K., and Northern H. T .: Effects of p-Chloropiienoxyacetic Acid (ANP-235)], Presse 2.4Dichlorophenoxyacetic Arid on Clicks, Science Med 68:215-216, 1960. 108:479-460, 1948. donee o f toxicity in the second patient, whereas 3,600 m g w as definitely toxic. A total of 16,312 m g o f sodium 2,4-D w as adm inistered to the second patient. T h e re were no necropsy findings w hich could be attributed to toxicity o f 2,4-dichlorophenoxyacetic acid. ■ I I I ' Uj » Î •• !: i .! :i i ,i ! .. ! } r ■» 5 5 ? 4. r -1 ¡■.I >< i-::ce fit ■ :l:c sllb■ was due — . The In countries where public health has been longest in effective operation, and where natality has responded to the new pattern of living, the demographic structure has undergone profound change, with a shift of population to older groups. Thus over-iiopulation and ageing are new challenges no less demanding than some of the infectious diseases which are passing. Moreover, where public health has succeeded emphasis has shitted from infectious to degenerative disease, now equally prevalent; some due to real increase, but mostly a manifestation of demographic changes resulting in a higher proportion of old people. New hazards also have arisen as the pattern of life changes with "development” ; carcinoma of the lung can be related to excessive smoking and smoke-laden atmospheres; suicide, alcoholism, and supposeciy stress diseases are directly or indirectly nervous phenomena, the response of the human to the strains of a highly organized society. There is still much barring the way to Utopia. C. P haser EfcncxiifGTOX, in Hobson, W .: The Theory and Practice of Public Health, London: Oxford University Press, l'>6 l, p. 7. 1 11!" '»f 1 eviV.î S tilbury 5351 -Jo O 6 So s fi. m ' SV. DALGAARD-MIKKELSEN a n d EMIL POULSEN Department of Pharmacology and Toxicology, Royal Veterinary and Agricultural College, Copenhagen, Denmark T ails or Contînt » I. Introduction........................................................... 235 II. Inorganic herbicide*................................................................................................... 236 A. Arseni te»................................................................................................................... 226 B. Sodium chlorate...................................................................................................... 227 C. Sulphuric acid...........................................................................................................227 III. Organic herbicides......................................................................................................... 227 A. Chlorinated phenoxy-acids......................................................................................227 B. Chlorinated aliphatic acid* and their sodium salts..............................................231 C. Carbamates and ally! alcohol..................................................................................234 D. Substituted urea*..................................................................................................... 236 E. Triasines................................................................................. 238 F. Substituted phenol*..................................................................................................340 G. Miscellaneous organic herbicides............................................................................ 241 1. Tributyl phosphorotrithioats (D E F )................................................................ 241 2. Maleic hydrazide................................................................................................. 243 3. EndothaJ sodium................................................................................................. 243 . 4. Diquat (Regione)................................................................................................ 244 5. Aminotriazole...................................................................................................; 244 IV. Conclusions.....................................................................................................................245 OOW517G99 TOXICOLOGY OF HERBICIDES I. INTRODUCTION In their endeavours to raise productivity, agriculturists have made use of chemicals to a steadily increasing extent. For the purpose of controlling injurious insects, fungi, weeds, etc., a great number of pesticides has been developed, among which the insecticides, especially the organophosphatc cholinesterase inhibitors, have attracted considerable toxicological interest. Relatively little has been written about herbicides, t.e., compounds which have been found useful for weed control. Many of the toxicological data underlying assessments of the risk involved by using them in practice originate from confidential, non-published reports placed at the disposal of the authorities concerned. Such data have not been included in the present survey. Of a number of compounds, the toxicology of which has l>ccn elucidated in relation to their use for purposes other than as herbicides, only a brief account will be given with references to relevant literature. Regarding the practical use of herbicides, reference is made to handbook literature (70, 80, 119, 120). Weedkillers usually are classified into two main groups, selective and non-selective. The selective substances can be used on crops without damaging the cultivated plants, whereas the non-selective types kill all vegetation. Substances of the latter group may therefore be used also for destroying potato haulm and for desiccation of green parts of plants which may unduly delay harvesting, in which case they are often called desiccants or de­ foliants. The herbicides are grouped, according to their mode of distribution in 225 5353 226 DALGAAUl>-M!KKKI.SEN AND FOtTLSEN the weeds, into contact herbicides, which are active only at the site of applica­ tion, translocated herbicides, which arc distributed throughout the whole plant from exposed parts of foliage or roots, and Tcsidual herbicides, which are spread on or in the soil and arc effective mainly against germinating seeds. For the present survey of the toxicology of the herbicides, a classification according to chemical configuration has been chosen. Inasmuch as common names, abbreviations, and registered trade marks * are used indiscriminately in the literature, we have, as far as possible, included the common names recom­ mended for pesticides by the British Standards Institution, marked (*), names approved by the British Weed Control Council (f), and those approved by the Weed Society of America (}). o O 3 cn •M o n. INORGANIC HERBICIDES Prior to the Second World War, mainly inorganic compounds were used for chemical weed control. However, as their actions are not very selective and they are often very persistent in the soil, so that damage to cultivated crops has been difficult to avoid, they have been replaced by organic compounds to a steadily increasing extent. Yet, various familiar inorganic compounds are still used as herbicides, e.g., calcium cyanamide, cupric sulphate, ferrous sulphate, mercurous chloride, potassium cyanate, and sodium tetraborate (borax). The toxicological data on these are well known from the handbook literature. The same is true of arsenites, sodium chlorate, and sulphuric acid, but these compounds will nevertheless by described briefly, because their use as herbicides has introduced special problems of toxicity. A. Arsenites Arsenites are used as non-sclcctive herbicides, especially for destruction of potato haulm, as aqueous solutions of potassium-ortho-arsenite (K*As*Oi) and sodium ortho-arsenite (Na»A8»Oj). The preparations may also contain metaarsenite and pyro-arsenite alkali salts. In rats, the LD50 for alkali arsenites has been found to be 70 mg/kg when administered by mouth and 150 mg/kg when applied dermnlly (32). In domestic animals (40) and man (105) the toxicity is considerably higher, fatalities having occurred in the horse, cow, and man after oral ingestion of 2 to 10 mg/kg. The solubility, preparation, and purity have a considerable influence on the toxicity of arsenic compounds (48); the toxic properties, actions on the animal organism, manifestations of poisoning, etc., have been excellently summarized recently (108). The use of onenites as weedkillers has occasioned an extensive series of acute, often fatal cases of poisoning in domestic animals, especially cattle, which have eaten contaminated crops or residues of sprays (40). Concentrations toxic to man have been found in milk from cows fed on crops contaminated by arsenites (61). Cases of direct human poisoning have also been observed. An instance has been described where one woman died and four others fell ill after drinking water from a reservoir which had been contaminated through a leaking f>ump valve in the 5354 0002881 227 tan k of an a p p a ra tu s for spraying arscnitc weedkiller. T h e drinking-w ater contained 180 p.p.m . (p a rts p er million) of As (7, 8 ). T his instance of poisoning in Great. B ritain resulted in a v o lu n tary agreem ent lwtwccn industry an d th e authorities for discontinuing th e use of arscnitc herbicides (32). B . Sodium chlorate Sodium chlorate an d o th e r chlorates arc extensively used for killing all vegeta­ tion of farm yards, railw ay track s, roadsides, etc., as well as for destruction of p o tato haulm . C hlorates possess no p articu larly great acute toxicity, th e LD 50 for sodium chlorate adm inistered by m o uth to ra ts having been sta te d to be 12 0 0 m g /k g (32). N evertheless, oral ingestion of both sodium chlorate and potassium chlorate, used in oral hygiene, has caused num erous coses of poisoning in m a n (21, 33, 105). In dom estic anim als, too, cases of acute poisoning have been ob­ served a fte r oral ingestion of chlorates (40). T h e m echanism of th e poisoning— m cthcm oglobin production a n d its consequences—has been well studied (21, 33, DOW 517701 TOXICOLOGY OF HERDICIOES 100). A special problem is th a t of th e high inflam m ability of sodium chlorate. T his can be reduced in com m ercial p reparations b y adm ixture w ith calcium chloride or sodium chloride, b u t a fte r dissolution in w ater an d spraying, dried residues m ay become ignited. In D en m ark , a tra c to r d river whose clothes had become im pregnated w ith sodium chlorate d uring spraying, died of severe burns after th ey were ignited by a red-hot exh au st pipe. C. S u lph u ric acid Sulphuric acid, which in certain countries, e.g., G reat B ritain, is com m only used for p o ta to hau lm destru ctio n , is often applied in high concentrations. Accidents w ith severe skin b u rn s a n d eye burns m ay therefore occur (32). In ­ halation of sulphuric acid m ay also exert a toxic effect, as indicated by studies on guinea pigs (3, 4) an d on v olunteer h um an subjects (5). n i . ORGANIC HERBICIDES A . Chlorinated phenoxy-acide C hlorinated com pounds of phenoxy-acetic acid, propionic acid, an d butyric acid, h av e been very extensively used w ithin th e p a st 15 years os herbicides under th e com m on nam e of "h o rm o n e w eedkillers." T h ey stim ulate p a rts of susceptible p lan ts, p articu larly w ithin th e group of dicotyledons, to excessive, uncontrolled grow th, w hich causes th e p lan ts to die (109). T h e m ost im p o rtan t com pounds, which in th e form s of w ater-soluble salts o r lipoid-soluble esters constitute th e activ e com ponents of com m ercial preparations, are listed in Table 1 , where th e L D 50 values in ra ts are also given. I n addition to th e com­ pounds listed in th e tab le, p-chlorophenoxyacetic acid (4-C PA ), sodium 2-(2,4(iichlorophcnoxy)ethyl su lp h ate (2,4-D E S-sodium , Scsone, SE S), and y -(2 ,4 ,5 trichlorophenoxy)butyric acid (2 ,4 ,5 -T B , 4 -(2 ,4 ,5 -T B )) are used as herbicides of this ty p e. 5355 0002832 228 DALGAAItD-MIKKKLSEN AND POUL8EN TABLE 1 Chlorinated phenoxy-acidt o O o -si ro 5356 00028G3 TOXICOLOGY OF HERBICIDES 229 DOW 517703 T h e nciilo toxicity following oral adm inistration to a num ber of experim ental anim als is m oderate. T h e LD50 as a rule is of the order of 300 to 700 m g per kilogram for the species examined (89), excepting dogs, which seem to be m ore susceptible, where the LD50 for both (2,4-dichlorophcnoxy)acetic acid (2,4-D ) an d 2,4,f>-T has been found to be 10 0 m g A g (27). E xperim ents w ith oral ad m in istratio n of various salts and esters of 2 ,4 -D as pure chemicals and as com m ercial preparations showed no significant difference in toxicity to a num ber of small anim als from th a t of th e free 2 ,4 -D acid (5 3 ,8 9 ). S hort-term studies on ra ts revealed no signs of reduced in tak e of food or inhibition of grow th in response to ad m ixture of 2 ,4-D a t 400 p .p.m . in th e fodder for 30 days o r 1000 p.p.m . for 14 days. Subcutaneous injection of 50 to 100 m g 2 ,4 -D p er kg to mice d aily for 90 days had no dem onstrable effect on th e general condition, fertility, o r tissue histology (53). In T able 2 arc recorded th e results of feeding phcnoxy-com pounds to larger anim als. E xcept for th e ra th e r consider­ able toxic effect on sheep of 2 ,4 ,5 - T P , adm inistered in th e form of propylene glycol butyl e th e r esters, these experim ents showed th a t th e large herbivores seem to tolerate prolonged ingestion of phenoxy-com pounds to th e same extent as ra ts. In dogs, on th e o th er h an d , these com pounds were found to have a pro­ nounced toxic a c tio n : 20 m g A g given fo r 2 to 3 weeks produced severe, fatal poisoning. P aren teral an d oral adm inistration of toxic doses of 2 ,4 -D to experim ental anim als brings about a characteristic complex of signs and sym ptom s (1 8 ,2 7 ,5 3 ), which has been studied p articularly in dogs. A fter a few hours th e anim als display a disinclination to m ove. T his passiveness is gradually aggravated, and a picture of m yotonia develops w ith rigidity of the skeletal m uscles and ataxia. T he condition m ay im prove transitorily w ith m ovem ent. In severe cases the anim als show progressive a p a th y , depression, and m uscular w eakness, especially of th e hindlegs, w ith periodic, clonic spasm s, and, finally, coma. T h e m uscular signs are accom panied by m arked anorexia; frequently, irritatio n of th e nose and eyes is indicated by scratching reactions. F u rth e r, bleeding from the nose and m outh m ay occur, as well os diarrh ea w ith blood-stained stools. Local irritation of th e alim entary tra c t often causes vom iting. H owever, this sign m ay be absent, even a fte r oral adm inistration to dogs (27). A utopsy m ay reveal necrotic ulcers of th e oral m ucosa and signs of irritation w ith histologically dem onstrable inflam­ m ato ry changes and necrosis of th e small intestinal m ucosa, as well as focal necrosis in the liver (27, 53) an d degeneration of the renal tubules. W hen given by m outh to dogs, even in fatal cases 2 , 4 ,5-T h a s produced only weak signs in th e form s of atax ia an d stiff m ovem ents of the hindlegs (27). As for the hum an response, a rep o rt is available (9, 75) of a m an who in a self-experim ent consum ed 500 m g of 2 ,4 -D daily for 3 weeks, w ith no perceptible effect. A ease of acu te fa ta l poisoning has been reported from D enm ark, where a young farm w orker com m itted suicide by oral ingestion of n o t less th a n €500 mg of 2 ,4 -D . W hen found, th e body showed signs of h aving been subject to violent convulsions before th e occurrence of d eath . T he changes post m ortem were unspecific hyperem ia of th e lungs, liver, an d brain (78). 5357 0002SC4 230 DALGAARn-MIKKELSF.N AND POOL8EN Daily Dost. — Animal Specie* Body Wright »«As Xnult MCPA Cow 30 Tolerated without detectable symptoms during 21 days (24) 2,4-D Sheep 100 (85) Dog 2-10 — Dog 20 Tolerated without detectable eymptome during 35 days Tolerated without detectable eymptome during 90 days 3 of 4 animals died, 18th to 49th day 2,4,5-T Sheep 100 Dog 2-10 Dog Sheep 2,4,5-TP Ref. No. (27) ÜOW517704 VV TABLE 2 Chlorinated ■phr.noxy-aeitia; xhart-term itludit» in dog, ekeep, and etna (27) (85) 20 Tolerated without detectable eymptome during 35 days Tolerated without detectable symptoms during 90 days 4 of 4 animals died 11th to 76th day (27) 100 Lethal after 11 doses (85) (27) The action of ingested phcnoxy-acid herbicides on muscular function, which is reminiscent of that following administration of halogenated acetic acid com­ pounds (22, 23), suggests an interference with carbohydrate metabolism. Two cases of a transitory diabetiform condition observed in spraying personnel following work with chlorinated phcnoxy-acid herbicides point in the same direction. However, hyperglycemia and glycosuria could not be reproduced with certainty in rabbits; only one out of live animals responded in this way in ex­ ploratory experiments, where the doses ranged from 125 to 500 mg/kg and the period of administration from fi to 50 days (68). The results of a long .series of investigations into the toxicity to domestic animals and game of herbicidal preparations and treated crops suggested that acute poisoning caused by consumption of crops from sprayed fields is unlikely to occur (16,24,46,75,87), excepting that "grass-eating” dogs may be poisoned by newly sprayed lawns. Human beings and domestic animals are. presumably liable to poisoning only by oml ingestion of highly concentrated preparations or spray solutions. A special problem in connection with the use of chlorinated phcnoxy-acid compounds is the tendency of the preparations—even in extremely low concen­ trations—to impart to water, milk, and other nutrients a very persistent chlorophenol-like odor and taste. Spilling of highly concentrated preparations close to wells or water courses has in several instances, by percolation through the strata, given a disagreeable taste to the water for long periods afterwards. Administra­ tion of 2,4-D to dairy cattle does not seem to result in excretion of Biologically demonstrable amounts in the milk (75). 5358 00028G5 3 TOXICOLOGY OF HERBICIDES 231 B . Chlorinated aliphatic acids an d their sodium salts DOW517705 Very few toxicological d a ta arc available regarding o ther chlorinated aromatic acids, introduced alone, o r in com bination as pre-emergence and selective herbi­ cides for agricultural crops. 2,3,6-T richlorophenyl-acctic acid (fen aej), 2 ,3 ,6 trichlorobenzoic acid ( 2 ,3 ,6 - T B A tt, T B A , T C B ) and 3-am ino-2,5-dichlorobenzoic acid (am ibenfl are used for these purposes. T h e LD 50 of 2 ,3 ,6 -T B A a fte r a single oral dose to ra ts was sta te d to be w ithin th e range of 700 to 1500 m g /k g (32). T h e chlorinated aliph atic acids possess m ore general phytotoxic properties. Some of these arc used, therefore, for defoliation and desiccation of cultivated p la n ts prior to m echanical harvesting. T he m ost im p o rtan t com pounds are: sodium trichloro-acetatc (T C A -ft sodium ), 6odium monochloro-acctate (SM CA), an d sodium -dichloropropionate (D aJapon-sodium *tt), as well as th e combined com pound, 2-(2,4,5-trichlorophen o xy)-ethyl-,2',2'-dichloropropionate (E rb o n f t) . j Studios on th e acu te toxicity of sodium trichloro-acetate have shown th a t this substance is not v ery toxic (118), th e L D 5 0 a fte r oral adm inistration being 3320 m g /k g for ra ts and 4970 m g /k g for mice. T h e anim als quickly go into an onesthesia-like state, which lasts for 36 hours, to be succeeded by coma and death, or by aw akening an d survival. Sodium trichloro-acetate has a far less localirritatin g action th a n th e free acid, which can corrode the skin an d mucous m em branes. S odiu m monochloro-acetale (SM CA) is considerably m ore toxic. I ts action on bacteria a n d in th e anim al organism is rem iniscent of th a t of o th er monohalogensu b stitu te d acetic acid com pounds, such as monoiodo- and monobrom acetic acid, th e enzym e-inhibiting an d bacteriostatic properties of which have been thoroughly studied (vide 22). T h e L D 50 a fte r oral adm inistration has been found to be 76 m g /k g for ra ts , an d 80 m g /k g fo r guinea pigs (118), and of th e same order fo r geese (20), w hereas fo r mice th e toxicity is som ew hat lower, th e LD50 being 255 m g /k g (118). I n th e sm all experim ental anim als ap a th y an d loss of weight w ere noticed, an d in th e fa ta l cases d eath occurred w ithin 3 days. The fatal oral dose for young c attle was found to lie w ithin the range of 100 to 150 m g/kg. Colicky-like restlessness an d incoordination were observed, developing in the course of 4 to 5 hours into universal fascicular tw itchings, gnashing of the teeth, an x iety , dyspnea, an d tachycardia. C om a an d d eath followed after 9 hours (23). T h is p icture, which is p robably referable to blocking of oxidative m etabolic processes, is rem iniscent of th a t seen in pigs (22) a n d dogs (6) poisoned with m onobrom acetic acid. T h e gross a n d microscopic findings a t au to p sy were likewise identical w ith those described fo r m onobrom acetic ocid-poisoncd pigs (22). Sodium 2 ,2-dichloropropionate (D alapon ) h as become of p articular interest owing to its hcrbicidal actio n on m onocotyledons, such os grasses, in relatively small q u an tities, therefore being useful as a selective herbicide in certain crops of cultivated p lan ts. D alapon is absorbed by p lan ts an d th en translocated. On this 5353 0002SG6 TABLE 3 Carbamate» — m DALQAAIID-MIKKEL 8 EN AND POULSEN Ül o ccn studied from several aspects through a com­ prehensive series of experiments (81). Administration of single doses to a number of small experimental animals has given LD50 values for rats, mice, guinea pigs, rabbits, and chickens within the range of 4000 to 9000 mg/kg. Two young cattle survived 1000 mg/kg given by mouth daily for 10 days. Beyond transitory symptoms in one of these (anorexia, diarrhea, indisposition), no signs of a toxic action were found by clinical or pathological examination. After oral administra­ tion of 15,50, and 100 mg/kg daily to dogs for one year, blood and urine analyses, as well as liver function, tests and histological examination of tissues revealed no signs of a toxic action, beyond an increased weight of the kidneys following the dose of 100 mg/kg. Examination of tissue specimens from the dogs showed up to 78 p.p.m. of Dalapon in kidney and liver tissues (81). Feeding of rats with Dalapon for 2 years in doses of 100,300, and 1000 p.p.m. in the fodder, corresponding to about 5, 15, and 50 mg/kg daily, was tolerated with no signs of a toxic action, apart from a minor increase in kidney weight on the largest dose. In these experiments with 1000 p.pjn., 10 to 30 p.p.m. of Dala­ pon was found on chemical analysis of the liver and kidney, and 20 p.p.m. in milk. Thus, cumulation docs not take place. In rats, reproduction and lactation proved to be uninfluenced through three generations with daily administration of 300 to 3000 p.p.m. of Dalapon in the fodder (81). The local-irritating action of Dalapon has been studied on rabbits, the skin of which was exposed daily for 10 days to a 10% aqueous solution. No more than signs of a mild, transitory irritation was seen (81). C. Carbamates and aüyl alcohol Within this group of herbicides, which, as shown in Table 3, comprises carba­ mates, thiocarbamatcs, and dithiocorbamatcs, is a number of compounds which are used especially as pre-emergence or selective herbicides on certain crops. Propham and chlorpropham have also been used to prevent potatoes intended for consumption from sprouting while stored. The toxicity of these compounds with prolonged ingestion is therefore of particular interest. Propham (0-/sopropyl N-phcnyl carbamate, IPC). Various investigations have shown th at carbamate estera possess a carcinogenic action. Thus intraperi­ toneal injection of isopropyl carbamate raised the frequency of lung tumors in a strain of mice from 17 to 90% (06). The idea of inquiring into a possible carcino­ genic action of Propham, which splits off aniline by acid hydrolysis, therefore suggested itself. However, with prolonged oral, intramuscular, and intrapleural administration of Propham to rats and mice, no signs of a carcinogenic action were found (56). This is in agreement with the results of experiments on groups of four rats each, given 400,800, and 1600 p.p.m., respectively, daily in the fodder for 3 months. No signs were demonstrable here of a toxic action on the general condition, growth, or fertility, and no pathological changes were seen a t autopsy or on histological examination of the tissues (101). 5362 00028G9 TOXICOLOGY OF HEIIIUCIOF.S 235 DOW517709 Experiments with the chemically closely related Chltrrpropham (tsopropyl-N[3-c.hlorophcnylJ carbamate, CII’C) have shown it to have a very low acute toxicity to rats and rabbits, LD50 by oral administration having l)ccn found to l>c of the order of 5000 mg per kg for both species (115). In feeding experi­ ments on male rats given 310 to 20,000 p.p.m. for 90 days, no effect was observed on growth. Doses of 1250 p.p.m. and higher produced an increase in weight of their livers, without histological changes being demonstrable (116). Jn the light of the above investigations, the observation of the development of skin tumors following painting of the back skin of Propham- and Chlorprophamfed rats with croton oil is of considerable theoretical interest (34). However, comprehensive feeding experiments (67) with administration of 2000 p.p.m. through 2 years to rats and 1 year to dogs revealed no carcinogenic effect. Feeding a t 20,000 p.p.m. provoked in both species signs of a toxic action, which was manifested in rats by retarded growth, increased mortality of the male animals, and increased liver and kidney weights, though without demonstrable histological changes. In dogs, retarded growth, increased weights of liver, kidney, and spleen, and splenic congestion were noted (67). Barbanc (4-chloro-2-butynyl-N-[3-chlorophenyl]carbamate) displays a some­ what greater acute toxicity than the Propham compounds, the LD30 after oral administration having been found to be 600 mg per kg for the rat and rabbit, and 2-10 mg/kg for the guinea pig. Dermal application of 1600 mg/kg over a period of 24 hours caused no deaths among rats. Oral administration of 9, 19.5, and 37 mg/kg daily for 22 days produced no toxic reaction, whereas 75 mg/kg over the same period effected loss of weight. Feeding experiments with rats showed no toxic action of 150 p.p.m. for 18 months (36). Barbane is a potent skin-sensitizing agent in man, in whom allergic reaction with rash develops at subsequent contact. Protection against cutaneous contact is therefore necessary during its use. Plastic (polyvinyl chloride) seems to be a more suitable protective material than rubber (36). SMDC (sodium N-mcthyl dithiocarbamate dihydrate, metham-sodium) has a special sphere of application, being used for killing weed seeds, soil nematodes, and the like by so-called soil sterilization. When applied in the soil it liberates gaseous methyltsoihiocyanaUs, the active substance, which is also available commer­ cially as an aqueous solution (Trapcx) for similar purposes. The LD50 of SMDC by oral administration to rats has been stated to be 820 mg/kg (29). The toxicity of mcihylisothiocyanate is considerably higher, the LD50 being 97 mg/kg (82). The main toxicological interest attaches, however, to the pronounced locally irritating action of these substances on the skin and mucous membranes, espe­ cially those of the respiratory organs, as well as the possibility of absorption of toxic amounts by these routes. The LD50 of SMDC after dermal application to rabbits has been found to be 800 mg/kg (29), whereas daily rubbing of methylisothiocyanate in 10% ethanol for 9 weeks into rabbit ears gave no more than a weak reaction (82). AUyl alcohol. This unsaturated alcohol has the same range of application as SMDC and methylisothiocyanate; it is applied in solution to the soil, where its 5363 000289® 23C DALGAARD-MfKKELSEX AND POULSEN DOW 517710 action is strong, but of short duration, so that cultivated plants can bo sown about two weeks later and grow without the interference of weed seedlings. Allyl alcohol has a strong locally irritating action and can be absorbed through the alimentary tract, the lungs, and intact skin. Extensive investigations of its toxicity have been made on experimental animals. The early literature, to which Miessner (74) contributed significantly by his investigations from 1891, has been summarized thoroughly (79). The toxicity and mechanism of poisoning have been studied in the United States within recent years (30, 65, 94, 95). LD50 values after oral administmtion to the mouse, rat, and rabbit ranged from 50 to 100 mg/kg, and after percutaneous administration to rabbits, from 45 to 90 mg/kg. Some time after the administration the animals were found apathetic, but not anesthetized, as they reacted strongly to pain stimuli. Lacrimation was seen, and often also increased secretion by salivary and gastroenteric glands, increased motility of the intestines, dyspnea due to pulmonary effusion, loss of weight, hemoconccntration, and vasodilatation. Autopsy revealed congestion of the organs as well as periportal necrosis (30,65). Local application to rabbit skin produced only transitory signs of irritation. Instillation into the eyes of rabbits resulted after 1 hour in redness and swelling, which were intensified in the course of 24 hours. In some instances keratitis was seen, but the condition returned to normal within a week (30). In acute inhalation experiments on rats, half of the exposed animals died in the course of 1 hour a t an air concentration of 1060 p.p.m.; 165 p.p.m. for 4 hours or 76 p.p.m. for 8 hours gave the same result. Experimental volunteers exposed to air concentrations of 0.78 to 25 p.p.m. for 5 minutes one to three times weekly over a period of 50 days, developed no signs of pulmonary dis­ comfort or affection of the CNS. The subjects could barely detect 0.78 p.p.m., while irritation of the nasal mucosa was first noticed a t 6.25 to 12.5 p.p.m., and eye irritation at 25 p.p.m. (30). Acrolein (acrylic aldehyde), the aldehyde corresponding to allyl alcohol, has been used for control of water plants and algae by introduction under water. The substance has the disadvantage of being highly toxic to fish, but this seems to be compensated for by its quick elimination. The LD50 after single 'subcu­ taneous administration is 30 mg/kg for mice and 50 mg/kg for rats. Moderate anesthesia occurs, as well as a few convulsive fits and dyspnea, especially in mice. Autopsy with histological examination of tissues has revealed pulmonary edema, chiefly perivascularly and hyperemia, slight fatty degeneration of the liver, and focal inflammatory processes in the kidney. At poisoning after inhala­ tion, where 0.3 p.p.m. for 30 minutes has a lethal effect, the histological changes— edema, hyperemia, and epithelial cell degeneration—are confined to the lungs (95). D. Substituted ureas The most important compounds belonging to this group of herbicides have been listed in Table 4, in which it is shown also that these compounds have a low acute toxicity when administered by mouth to rats. For 3j(p-chlorophenyl)-l, 1- 5364 00028Si TOXICOLOGY OF HEKDICIDKS 237 DOW 51771Ì TABLE 4 Subetituted ureae dimcthylurca (Monuron) the approximate lethal dose by single oral administrat ion to guinea pigs and rats has been set a t 670 and 1500 mg/kg, respectively (28). To the same group belong also N-butyl-N'(3,4-dichlorophenyl)-N-methylurea (W huronft), 1 ,3 - d i(2 ,2 ,2 - trichloro-1 - hydroxycthyl)urca (Dichloral urea, D< '(JJ, DU), and N-cyclooctyl-NW-dimethylurea (OMU). The sulwtituted urea derivatives destroy numerous species of plants, and their activity persists for a long time. They ore used in large doses for eradication of all vegetation of farmyards, railway tracks, roadsides, etc. They may also be used in much lower concentrations on cultivated crops, such as asparagus. Owing to the long persistence of these substances, residues may be present in harvested crops. In feeding experiments on rats given Monuron at 25, 250, and 2500 p.p.m. for 2 years, no effect was observed on survival or frequency of tumors, as compared with control groups and in the colony of normal rats. The conclusion was drawn that “there was no indication that Monuron is canccrogenic” (54). In the group given 2500 p.p.m. the mole rats were seen to lose weight after only one month of feeding. During the experiment mild anemia was also found, and a t the end the 5365 0002892 238 DALGAAIMi-MIKKELSEN AND POULSEN OOW517712 weights of the liver and spleen had increased. On histological examination of tissue specimens none of the groups presented changes attributable to an action of Monuron. In experiments on dogs, which received in their diet 2.5,12.5, and 25 mg Monuron per kg for one year, no signs of toxicity were seen during the ex­ periment, and a t autopsy with histological examination of tissue specimens no changes were noticed that seemed to originate from feeding with Monuron (54). Bats given 500 p.p.m. of Fenuron in the diet for 90 days tolerated this dose with no signs of poisoning (91). Diuron was likewise tolerated a t 50 p.p.m. While 5000 p.p.m. of this compound for 90 days caused no deaths, but loss of weight, a fall in the number of red blood corpuscles, and pathological changes in the spleen were noted (28). Preliminary experiments using Monuron and Diuron on guinea pigs have shown no signs of allergic skin sensitization. Neither have such reactions been observed in experimental subjects or factory employees working with these com­ pounds (28). E . T riazinea The announcement of the phytotoxic and plant-growth-regulating properties of a series of aminotriazines (41), especially the 2-chloro-2,4-bis(ethylamino)1,3,5-triazine compound, Simazine, initiated the synthesis of a great number of herbicidal derivatives. The two most commonly employed of these are shown in Table 5, whereas the compounds known under the names of: Propazine }(2-chloroTA B LE5 Triazinti Cnwwna Fenaala Caapaud S-CUoro-4,8-bit (tlkylamino)-l ,S,Strianru Sima- Cl MOO (42) 1750 (43) *MWt t I V H LDJ0. HK/kf bf Patent M . Aamisiatn- Ne. tie* to Rett \ H NN -l A-l/ / \ / \ CHtCHi N CHt—CHa t-Chloro-4-ethylamino S-isopropylaminot,3,S-lriatin* Cl A in sum! l H CH, \ \ 4 H ^C / / CH, H A V '''C H i—CH, * _____ 5366 0002803 TOXICOLOGY OF HERBICIDES 239 DOW 517713 TABLE 6 4,6-Ws[tsopropylaminoj-l ,3,5-triazine), Trictazineî (2-chloro-4-diethylamino6-cthyIamino-l,3,5-triazine), IpazineJ (Isodiazinc, 2-chloro-4-diethylamino-6wopropylamino-1,3,5-triazine), Simetoncf (2 ,4-5w[ethylamino]-6-methoxy1,3,5 - triazine), PromctoncJ (2,4 - 6ts[t$opropylamino] - 6 - methoxy - 1,3,5triazine), and Atratoncf (2-ethyiamino-4-wopropyIamino-6-mcthoxy-l ,3 ,5-triazinc), are still being used mainly experimentally. These compounds are almost insoluble and persist for a long time in the soil. They have been used mainly for eradication of all vegetation as well as for local wcodkilling under fruit trees and shrubs. Certain species of plants, e.g., asparagus. and maize or com, arc, however, resistant to the hcrbicidal action of the triazincs. l'hoir use on areas with these cultivated plants is therefore practicable but involves a risk of residues in crops for consumption. Studies on the toxicity of Simazine (42) showed that the LD50 after oral ad­ ministration to mice, rats, rabbits, chickens, and pigeons in all cases exceeded '*000 mg/kg, whereas all rats survived daily doses of 2500 mg pure Simazine per kg for 4 weeks, and 1250 mg of a commercial preparation per kg. R ats fed for two ycara on a diet containing 1,10, and 100 p.pjn. of Simazine revealed no signs of 5367 0002894- 240 DAI.GAAHD-MIKKELSKN AND POULSEN F. Substituted phenols DOW 517711 toxicity (40a), and screening test* for various acute actions, such os spasmoiysis on the isolated intestine and analgesia in mice, did not demonstrate significant activity (40a). The LD50 of Atrnziue (‘J-chlnro-4-ethylamino-6-tsopropylamino-l ,3,5triazinc) (43) after a single oral administration was found to be 1750 mg/kg for mice and 3080 mg/kg for rats, while a daily dose of 400 mg/kg through 6 weeks proved fatal to half of the test rats. Dinitrophenols. The checkered history of the dinitrophenols also includes a chapter on their applicability as herbicides. In agriculture these compounds are used also as insecticides, ncaricidcs, and ovicides, for potato haulm destruction, pre-harvest desiccation of leguminous seed crops and apple-thinning sprays. The chances of contact are therefore great, and their users are often poisoned when protective measures are disregarded. Three derivatives are commonly used: DNOC (DNC, 2-methyl-4,6-dinitrophenol), Dinoscb (DNPB, 2-sec.butyl-4,6-dinitrophenol), and Dincx (DNOCIIP, 2-cyclohexyl-4 ,f>-dinitrophenol). The chemical configurations and LD50’s for rats after oral administration are shown in Table 6. The mechanism of toxic action which is based on the uncoupling of oxidative phosphorylation, has been studied in detail; a survey has been given in this journal (17). A great num­ ber of toxicological data have been given in Handbook of Toxicology (77), and poisoning of man and domestic animals has been reported in recent surveys (12, 32, 40, 72, 80, 88). Clinical accounts are available from Belgium (52) and Great Britain (15), among others. Up to 1950 seven British cases had been re­ ported of fatal poisoning with DNOC used as a weedkiller. Countermeasures against poisoning (14) have since been satisfactory in Britain, no fatal accidents having occurred aftertheir introduction (12,32). Peniaehlorphenol. Like the dinitrophenols, pentochlorphcnol is an effective uncoupler of oxidative phosphorylation. The mechanism of toxic action has been studied in experiments with molluscan and mammalian tissues as well as on en­ zymes (110, 111, 112, 113). It seems to be due to inhibition of the intracellular transfer of energy-rich phosphates as well os their synthesis. In agreement with this, the symptoms of intoxication include acceleration of respiration, hyperpy­ rexia, hyperglycemia, glycosuria, and promptly occurring motor weakness. As in cases of dinitrophenol poisoning, rigor mortis sets in early and is very pro­ nounced (26, 45, 64, 71). The acute toxicity of pentachlorphenol as well as that of its sodium salt have been studied by oral, subcutaneous, and dermal administration of single doses to rats and rabbits (2G, 64, 71, 102), and by single subcutaneous injections into dogs (71). The LD50’s and the minimum lethal doses were found to vary considerably, ranging from 27 to 550 mg/kg. The highest toxicity was noted for pentachlor­ phenol dissolved in mineral oil (fuel oil) given by mouth to rats, and the lowest for the sodium salt administered in the same way to rabbits. After dermal appli­ cation, mineral oil solutions arc likewise more toxic than solutions in vegetable 0002895 TOXICOLOGY OF IIKliniClDKS 241 oil ¡iiul in wilier (64). Furthermore, the local reaction, which may manifest itself l>y changes from cryllicma to grave dermatitis, is stronger with mineral oil as vehicle ((>1,71). The results of short-term studies on a manlier of animal species suggest a certain degree of cumulative action, especially after subcutaneous injection (71); signs of such have also been seen after oral administration to rats and eats (26). In experiments on calves, on the other hand, cumulative action has not been ob­ served (51). From the pract ical use of pentachlorphenol, this compound is well known as a skin irritant (12); in addition, toxic amounts may be absorbed through the skin. In relation to its use os a timber-preservative, human coses of poisoning have occurred, some of which have been fatal (73,107). Sodium pcntachlorphenate in aqueous solution with mineral oil added, used as a herbicide in pineapple plantations in Australia, has been described as the cause of poisoning of nine workers, five of whom died. In one, who died after 21 hours, pentachlorphenol was demonstrated by chemical analysis of tissue speci­ mens in quantities of 2 to 14.5 mg per 100 g tissue (45). In cattle, coses of poisoning have been described which ran a fatal course within 24 hours of oral ingestion of 5% pentachlorphenol in kerosene (97). Mild signs and symptoms of poisoning in sheep and cattle have followed 25 mg/kg (85). o O £ CJl r-* Cl G. Miscellaneous organic herbicides 1. Tributyl phosphorotrithioate (DEF). Organophosphates have become very extensively used os insecticides within agriculture and horticulture. A compre­ hensive literature is available on the toxicity of these compounds, related to their cholinesterase-inhibiting action (55). In studying DKF, which is used as a de­ foliant in cotton fields, attention has been focused on the relatioaship between the toxicity and the inhibiting action on the cholinesterase activity in different organ tissues (76). The LD50 after oral administration to rats is 325 mg/kg (Tabic 7). With daily intraperitoneal injections of 50 mg/kg into rats for up to 60 days no signs of poisoning were observed, although after sacrifice cholinesterase of brain tissue was found to be 14 % of the average in control groups. Injection of 100 mg/kg was lethal after 5 to 30 days. In acute poisoning experiments de­ pression and inactivity were seen after one hour. These symptoms persisted for 2-1 to 48 hours, unless irritability or manifestation of pain or tremor was produced by external stimuli. The depression developed in some eases into muscular weak­ ness with rigidity. Not till 2 to 0 hours after injection did the animals also display the usual signs of anticholinesterase poisoning: profuse urination, salivation, and lacrimation, and respiratory impairment increasing to respiratory paralysis and death (7G). The cholinesterase activity of the red blood cells and brain was often poorly related to the. cholinergic signs, often with freedom from any signs or symptoms despite a low activity level (76). Having licon observed also in cases of poisoning with other organic phosphorus compounds (39, 63), this fact does not militate against the hypothesis of cholinesterase inhibition os an etiological fac­ tor in the mechanism of poisoning. The somewhat differing manifestations of poisoning, and the failure of prophylactic and therapeutic effects of enormous 0002896 ( C fla p ú u d 8,8,8 -trib u tyl phoiphorotrithioal» ¡ ,3 ,3 ,8 -Utrabydro-S,8-diozopyridatine Ditodium 7-ozabicyclo-(3,3,1 )heptant-t,3-di• earbozylate, Diaodiura 3,ft-andoxohexahydrophth«Uta 9,t0-dihydro-8a, lOa-diazoniaphmantkrtn* dibromidi, l.J'-etbylene-a.a'-dipyridylium dibromida « en co O O o rj co <£) o 3-amino-t ,3,4-lriatol» TABLE 7 M ie c e lla n e o u t o rg a n ic h e rb ic id e » LD50, mffAfbr C ouM or Abbrcvltld Names Perotal Admia* auiioo to Hsu Fwmttli CH,—CH«—CH,—S B«(.So. D BF 325 (76) Maleic kydrazide, MH( 4000 03) Endothal -»odium 1t 80 (32) 400-440 (83) 25000 (90) CH,—CH,—C H . - 8 - P 0 1 C ch'^ \*h 1 NH 1 CH V i 0 H «/ H tC ^ I ^ C H —COON» 1 o 1 H,C 1 CH—COON» v ^H HC—CH HC^ HC—CH N C -C / D iiu o l/tt Region»* ^ C H , 2B r' llc— N^ \ +— CH H^C—cil, HN -- N hA A— NH, V Amitrole.t ATA, amtxol, •mitol S T i A T S M O a NBBtOOd Q N V NHSTiaUMlW-iUlVVDaVU CH.—CH ,—C H ,-B / A TOXICOLOGY OF HEIU1I0IDK8 243 DOW 517717 doses of atropine (70) suggest, however, that other facto» also contribute to­ wards t he toxic action of DKF. 2. Maleic hydrazidc (1,2,3,6-tctrahydro-3,6-dioxopyridnzine) is used for spraying on roadsides, hedges, and the like, to facilitate and reduce the work with cutting, etc. The growth-inhibiting properties are utilized further by spraying the compound on root crops 2 to 4 works before harvesting, as this greatly reduces the tendency to sprouting during storage. In experiments on a number of small animals, the acute and chronic toxicities of this, compound were found to be low. Thus, for instance, the LD50 after oral administration to rats was 4000 mg/kg, while prolonged feeding experiments on rats with 10,000 p.p.m. of maleic hydrazide in the fodder showed no signs of a toxic action within the normal life span (13). The demonstration of the fact that in Vicii faba roots maleic hydrazide not only inhibited the growth of cells, but also caused breakage of the heterochromatin in the chromosomes (25) aroused a great interest in ascertaining whether animal cells might also be susceptible to this action, or whether it is specifically directed against plant cells. This is im­ portant, because the ability of substances to produce chromosome breakage may be accompanied by carcinogenesis. In thorough investigations, comprising weekly subcutaneous injections into mice and rats of 500 mg/kg for 100 weeks, as well as feeding experiments over the same period with 10,000 p.p.m. of maleic hydrazide in the diet, no signs of a carcinogenic action were demonstrable. Tests for a possi­ ble co-carcinogcnic action on application together with croton oil to mouse sk in revealed no greater tumorigenic action than th at of the oil alone. Implantation of maleic hydrazide pellets in the bladder of mice did not give rise to tumor forma­ tion, and administration of 125 mg of the compound per kg to tumour-affected rats had no effect on the tumour growth or cell mitoses. In experiments with tissue cultures of mouse epidermis and guinea-pig skin, no effect was seen on cell divi­ sion, mitosis, or tissue respiration (13). The fate of maleic hydrazide has been studied in rabbits. After oral administra­ tion of 100 mg/kg, the rabbits excreted 43 to 62 % unchanged in the urine in 48 houra. Coupling products were not detected, and the fate of the remaining 40% of the administered amount could not be determined (13). 3. Endothal-sodium (disodium 3,6-endoxohexahydrophthalate), which has strong plant-damaging properties, is used particularly as a defoliant and desiccant in leguminous crops, and has also been tried as a pre-emergence herbi­ cide, e.g., for sugar beets. The compound, which is closely related to cantharidin, is very toxic. After oral administration to rats, the LD50 has been found to be 80 mg/kg (32), whereas the lethal dose by intravenous injection into rabbits and dogs is of the order of 5 to 10 m g/kg (44). Shortly after administration, scratch­ ing of the nose was observed, and in the dogs, intermittent retching and vomiting also occurred for 2 houra, followed by death or recovery. In the rabbits, head­ shaking and irregular, greatly accelerated respiration were noticed (98). For both animal species, death was attributed to respiratory failure. In experiments with isolated ventricles of frog hearts and rabbit atria functional depression was seen in response to a 1 in 20,000 solution of Endothal-sodium (44). Hence, cardiac de- 5371 0002SS8 244 c DAI.(»AAW)-MIKKELSR.V AND POULREN o o $ 517713 prcssion has been supposed to l)c tlm cause of death. However, intravenous in­ jection of Fndolhal into anesthetized dogs, atropinized os well as non-atropinized, caused a fall in blood pressure and respiratory failure, whereas changes of the ECG were first noticed in association with severe respiratory impairment in the terminal phase. While cats reacted in the same way as dogs, although with a less pronounced fall in blood pressure, domestic fowls were found to be insensitive to doses four times larger than those which were active on dogs and cats (98). The effects of Endothal both in vitro and in vivo were found to be characterized by manifesting themselves only after a long latency (98). This points towards a metabolite as the proper active factor. Application of a 10 to 20% aqueous solution to the skin produced erosions which developed into necrosis, in some cases resulting in death owing to cutaneous absorption (44). The LD-50 after dermal application to rats was 750 mg/kg (32). 4• Diquat (Reglone). Diquat (9,10-dihydro-8a,10a-diozoniaphenanthrene dibromide) has been proposed as an agent to produce desiccation and defoliation prior to harvesting, destruction of potato haulm, and as a pre-emergence and a total herbicide. The LD50 after oral administration to rats was 400 to 440 mg/kg, whereas the LD50 after subcutaneous injection was 20 mg/kg, with death occurring in the course of 5 to 7 days. At autopsy, the intestinal canal, especially the cecum, was distended; histological examination revealed signs of gastrointestinal irritation, and in the lungs, thickened alveolar walls, particularly after large doses (83). Intrapcritoneal injection of 500 mg/kg to rats produced cyanosis and convul­ sions, culminating in death after 2 hours. Subcutaneous injection of 1 mg/kg into rats for 3 weeks caused no Rigns of toxicity. Similarly, feeding experiments to rats over 14 months with .500 p.p.m. in the diet disclosed no carcinogenic or toxic action (83). Following oral administration of CM-labclIcd Diquat, 90 to 97% of the amount of activity given was recovered in the feces in the course of 48 hours (83). This, correlated with the difference in toxicity after oral and parenteral ad­ ministration, suggests minimal absorption from the gastrointestinal canal. 5. Aminotruuole (3-amino-1 ,2 ,4-triazole) is hcrbicidolly active against nu­ merous species of plants. After absorption, it is translocated through the root as well as through the foliage. I t breaks down rather quickly in the soil, and the exposed plants become chlorotic, as aminotriazole seems to interfere y ith chloro­ phyll production (47). In animal experiments, aminotriazole showed a low acute toxicity, the LD.50 after oral administration having been found to be 14,700 mg/kg for mice (114) and about 25,000 m gAg for rats (90). No signs of poisoning were seen after in­ travenous injection of 17.50 mg/kg to a cat (90), of 1600 mg/kg to mice (90), or 1200 mg/kg to a dog (114). A single intraperitoneal dose of 4000 mg/kg was tolerated by mice, whereas 21 doses of 1000 mg/kg each to rats, distributed over 45 days, produced an increase of the thyroid weightof 328% in males and 410% in females, while at the same time the growth and intake of food were found to bo normal (90). In feeding experiments on rats extending over 68 weeks, ino effect was seen on the growth or intake of food a t 10 and 50 p.p.m. in the diet, as compared with O u t tv 0002899 TOXICOLOGY OF HERBICIDES 245 DOW 517719 controls, whereas 100 p.p.m. caused a decrease in growth and food intake in male rats during the last few experimental weeks. However, in male rate given 50 p.p.m. the thyroid Income enlarged after 13 weeks (114). Investigations into the effect- on the thyroids of rats of 2 years of feeding with aminotriazolc am stated to have given the following results: In the control group, there was found one ease of cystic follicle with papillary changes, while among the animals given 10 p.p.m. one out of ten examined presented adenoma; further, after 50 p.p.m. two out of 15 examined had adenoma and one apparently adenocarcinoma. Hats given 500 p.p.m. of aminotriazolc in the diet for 17 weeks, which were then retained on a diet free of the compound for 2 weeks prior to sacrifice, appeared to have normal thyroids a t the time of sacrifice (62). The observat ion that rats fed with 100 p.p.m. of the compound for 2 years "developed a significant number of thyroid adenomas and adenocarcinomas,” as well os the demonstration of residues of aminotriazolc in marketed cranberries (31), caused prohibition of sale of cranberries and cranberry products of the 1958 and 1959 crops from certain parts of the United States. Furthermore, it resulted in an official announcement from the Secretary of Health, Education, and Wel­ fare that the reason for the prohibition was "possible contamination by a chemi­ cal weedkiller, Aminotriazole, which causes cancer in the thyroid of rats” (37). In the ensuing discussion, doubts were raised as to whether the marked anti­ thyroid action of aminotriazole can justly be characterized as carcinogenic (10, 62). The mechanism of the antithyroid action of aminotriazole seems, on the basis of experiments on rats, to be identical with that of thiouracil derivatives (la, 62). The absorption of I m by the thyroid is depressed in both rats (la) and humans (10), because its incorporation as organically bound iodine is obstructed (la). Following injection of aminotriazole, a reversible inhibition has been noticed of the catalase activity in thyroid tissue (la) as well as in kidney and liver tissues (49, 50, 104). However, to obtain a definite inhibition of the catalase activity, larger doses of aminotriazole are required than are necessary to depress the ab­ sorption of I m by the thyroid (la). Hence, inhibition of catalase activity can hardly account for the antithyroid action. Studies, in vitro, using purified catalase preparations from liver and red blood corpuscles, have under special experimental conditions shown irreversible inhibition (69). However, the results of experi­ ments with thyroid tissue suggest that inhibition of thyroid peroxidase is the essential factor (2), a hypothesis that is borne out by the results of experiments with peroxidase from milk and vegetable tissue, which are likewise inhibited by aminotriazolc (19). I t should be added, however, that, the question has not yet been clarified, since enzymes that are active in the purine metabolism of bac­ teria arc also inhibited, in these cases reversibly by aminotriazole; this observa­ tion is utilized experimentally (84,117). IV . C O N C LU S IO N S Apart from the fact that most information regarding the toxicology of the lierhicidcs is os yet rather fragmentary—a situation which will be considerably improved when reports from the biological laboratories of industry become avail- 00 0 2 9 0 0 246 DALGAARD-MIKKELSEN AND FOULSEN DOW517729 able to the public—it is important that attention not be confined to the toxicity of the original compounds. Their influence on plant metabolism might, for in­ stance, block synthetic chains, leading to the accumulation of endogenous toxic products in vegetables. In this respect it has been shown that 2,4-D, applied in subtoxic amounts to such plants as sugar beets, increases the concentration of nitrate to twenty times the normal level. Consequently, feeding with the foliage involves a risk of poisoning (tw). Attention has also been focused on the increase of cyanide in cyanogenic plants exposed to the action of herbicides (103). As to the toxicity of metabolites of the herbicides themselves, which are formed in the soil and plants after their distribution, little is known. The problem has been touched in connection with investigations into the breakdown of herbicides by the aid of the micro-organisms in the soil. In particular,'the decomposition of the chlorinated phenoxy-ncids and the chlorinated aliphatic acids by soil bac­ teria and fungi has been submitted to extensive studies (vide 11, 57, 58, 59, 60, 93,121). I t appears that the breakdown of these compounds is almost complete, involving dehalogenation (57-60) and hydroxylation of the side-chain (93). Therefore,^persistent deleterious effect on the microflora of the soil need not be feared after use of the great majority of these organic herbicides (38). This is probably due to the fact that in most cultivated soils the populations of bacteria and fungi are so rich that there will always be found a t least a few individuals that con adapt to almost any substance presented to thepij These will synthesize new specific enzyme systems using the foreign substance as substrate. The re­ sistant strains of micro-organisms will therefore multiply excessively, until the “substrate” has been decomposed (11) YIn general, the biotransformation of herbicides in soil does not seem to lead to the production of more potent compounds7(ll, 93, 121). Certain inorganic herbicides, e.g., chlorate, may, however, have a deleterious influence on bacterial nitrification in the soil. By reduction of chlorate, hypochlorite may be produced, which has a very strong inhibitory ac­ tion on the growth of the soil bacteria (1). Pentachlorphenol may have a harmful effect, especially on the soil fungi; various emulsifying and surface-active agents in the hcrbicidal preparat ions may also interfere with the soil flora (38). . The fates of the herbicides in plants have as yet been scarcely elucidated, but intensive studies are being carried on to clarify biotransformation in plants. The explanation of selective phytotoxicity has been founded on these studies;'Uccisive progress was gained when it was discovered that the herbicidolly inactive 4-(2,4-dichlorophenoxy) butyric acid is converted by beta-oxidation to the active 2,4-dichlorophenoxyncctic acid in plants where specific beta-oxidase systems are present. These plants arc harmed while species not possessing the specific en­ zymes are resistant (109). Conversely, the triazinc derivative, Simazin, in non­ sensitive plants is degradated by the presence of different factors such as catalase, peroxidase, and polyphenols to substances without phytotoxicity (46a) J Great difficulties in analytical procedures have to be solved before the fates of Jrerbicides in plants are clarified ^92). However,-there is reason to expect th a f c more rational basis will eventually be available, both for the study of the metabolism of herbicides in the animal organism, and for the planning of experimental in- 5374 0002901 TOXICOLOGY OF HERBICIDES 247 REFERENCES 1. Aaxno. B.:OntheBiechariiom of thetnxio action alohlertlea aadoamoraUtedoaholi rumoa youngwheat piaale. KiinxL LaotbrllAmk. Ana. 13:17-107, 1144. la.ALRXAnoca. N. M.: Antithyroid actios oil-amioo-1,2 ,4-triaiole. ). bW. Chom.234: 140-110, 11». 1 Alcxakor*. N. M.: Iodide peraxidaaa in ret thyroid And aelirary giando Aad iU in h ib it» by antithyroid noropouiuU. Fed. Proe. 14: bo. I, port 1 , 1 7 1 , 11». 'J. Axoon. M. 0.: The rexpiratoryiutponaaol guinea pirn toeulfurieaeid mint. Arab. iodoetr. Hlth II: 4*7-414, 1IU. 4. Annua. M. 0 .. SenuLt, R. Z. abb D n n t t , P.: Toxicity el eulfurie odd mint to nine» p i». And. iadiatr. lin t. 5: JIM ». 1132. I Annua. H. 0.. 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