Document 93b2B4d3VbNy9mRmjDqEJE3d5

k Y) --------------- > vK- ,a.!37a -C ic C i^ssJ . , I M * 0 7 fc - ' ' Journal of Occupational Medicine Official publication of AMERICAN OCCUPATIONAL MEDICAL ASSOCIATION and of AMERICAN ACADEMY OF OCCUPATIONAL MEDICINE OCCUPATIONAL ALLERGY TO LABORATORY ANIMALS: EMPLOYER PRACTICES To determine current American em ployer attitudes and policies regarding employee allergy to laboratory animals, questionnaires were sent to 155 institutions. Laboratory animal allergy was reported as a workplace disease o f animal house employees by 108 facilities (70% ), w ith rat and rabbit ex posure the most frequent cause. A uniform policy regarding the problem in o o U.S. animal facilities is not presently apparent. T THE RISK OF INFERTILITY AND DELAYED CONCEPTION XO oD ASSOCIATED WITH EXPOSURES IN THE DANISH WORKPLACE ---J >- .1 -- > o re -njj -'rjj jr z?< Data collected from medical records and mailed questionnaires were utilized in a case control study o f the association between in fe rtility and a num ber o f Occupations and occupational exposures. Results suggest that male exposure to heat, and female exposure to noise, textile dyes and lead, m ercury, and cadmium are associated w ith in fe rtility . * t: o VIBRATION WHITE FINGER DISEASE AMONG TREE FELLERS IN BRITISH COLUMBIA Symptoms o f vibration w hite finger disease (V W FD ) were present in 70% o f 146 men who had been engaged in tree felling fo r 11 to 15 years and in 75% o f the men who had been employed in this occupation fo r more than 20 years. There was objective evidence o f disease (delayed finger rewarming after cooling) in 31 o f the 4 3 fellers w ith symptoms (72% ) and in .13 o f 74 control (18% ) w ith o u t symptoms. The median latency period fo r fellers w ith sym ptom atic VW FD was 7.3 years. ^ PR0JCT10NS OF ASBESTOS-RELATED DISEASE 1980-2009r Among persons w ith nontrivial exposure to asbestos, it is estimated th at approxim ately 19,000 cases o f mesothelioma and 5 5 ,0 0 0 cases o f lung can cer w ill arise in the United States between the years 1980 and 20 0 9 , and th a t there are approxim ately 6 5,000 U .S. men now alive w ith clinically diagnosable asbestos. The cancer estimates are based on the number o f asbestosexposed workers required to produce the current national incidence o f m esotheliom a. rs> cn as r*o co co 4700 May, 1983, Volume 25, No. 5 'j ) - 0009692? J Letters to the Editor Rcoders arc invited to submit letters fo r pub lication in this department Submit to Doris Flournoy, Publisher, journal o f Occupation al Medicine, 1845 W. Morse, Chicago, IL 60626. Letters should be typewritten, double spaced and should be designated " For Publication. " Herbicide Exposure and Pulmonary Disease To the Editor: Exposure to the herbicides Karmex Idiuron) and 2.4-D has not been associated with signifi cant pulmonary disease. This report describes an unusual exposure which took place in 1982. It is a common practice in the rail road industry to reduce the foliage along the tracks by spraying herbicides such as 2,4-D or Karmex. During the spring o f 1982. two healthy young men, an engineer and conductor, re ceived an intense exposure to these herbicides. The train was being driven into a strong head wind while pushing tank cars containing the herbicides in fro n t o f the locomotive. Significant amounts o f the spray were whipped by -the wind into the locomotive as it pushed the spraying u nit along. The herbicide spray opacified the front windshield o f the locomotive, making it constantly necessary for the en gineer and conductor to lean out o f the side windows in order to drive the train. After the second day o f the spray exposure (the day when the train pro ceeded into the head wind), both patients noted symptoms o f itching and burning involving their oral and nasal mucosa and their conjunctiva. In areas where their skin had sweated or had made contact with glasses frames, small ulcerations appeared. Within 24 hours both men developed significant chest discomfort with cough, which was initially productive o f a mucoid sputum. They complained o f mild headache, some muscle tw itch ing, and throat soreness. Within the next 4-5 days, the cough and sputum production decreased, but both pa tients continued to feci somewhat weak and dvspncic. Initial complete blood counts were normal as were chest x-rays. Subse quent pulmonary function tests re vealed no abnormalities o f air flow, 4 i ' f ? I t \ i iI L if. . j .. **..,,.,ijL if* *.-* VC' 2 V 7 3 . '(too) O' , K * I *2 {p I; lung volumes, or diffusing capacity. Both men have continued to feel that their health has been impaired, al- though their complaints are non- specific. Both are concerned 'about the possibility o f long-term pulmonary toxicity. I believe that the exposure o f these men to 2.4-D and Karmex may have been more intense than exposures previously reported. Factors such as the duration o f exposure, particle size o f the aerosol, and biological proper ties o f the vehicle may have been o f importance. The manufacturers o f these products have been unable to provide adequate information. 1 would urge any Journal readers who have dealt with similar problems io publish or communicate their ex- periences. . - - Kennerh G. Tonington, M.O. 425 W. Bannock Si. Boise. 10 83702 o o z rvi -- c_n CT5 CO f | < -J \ \ l ! ; * | 4701 ^ -n h r 000968: 4702 MATcH IALJNVOLVED:. SOURCE OF INQUIRY: _ STREET ADDRESS:____ CITY:______________ RETURN CALL TO: to * * s~ PHONE: 1 7 5 3 ) NATURE OF REPORT (NAME - DATE - PLACE): // j2*lS . 'tP sL ^ ju * ^ 0 ? - < ? < *? -- & S T A / f g^. **. M -CD . Ok g ^ -/A / / REMARKS ON FOLLOW UP: O >0 rsa 0008368 'P - H fo e t ,J- *-.a ':i I v*. :.->  -m ifro ' OOH2I579I6 DISTRIBUTION Adcock, L. 0 ., 2020 Dow Center A rrington, J. P., 9008 Bldg. Baldwin, M. J ., Sarnia Ballantyne, G. D., Hong Kong Bjerke, E. L ., 9001 Bosatra, 0 ., Milan, Ita ly Caputo, R ., Sao Paulo Chen, U. L ., Sarnia Corson, F. P., Coral Gables D ietz, F. K., Lake Jackson, TX Ehrmantraut, J. VI., 9008 Bldg. Engibous, 0. L ., B-1210, Freeport Frber, H. A ., 2020 Dow Center Flynn, J. P ., Larkin Lab Gehring, P. J ., 2030 Dow Center Gum, U. F ., 9001 Bldg. Hinze, C ., Lake Jackson, TX Hoerger, F. D., 2030 Dow Center Holder, B. B ., 2030 Dow Center Jersey, G. C., Lake Jackson, TX Jones, J r . , I . C., Plaquemine, LA Koppleman, H., Stade Lancini, G., Milan, Ita ly Lanham, J. M., Sarnia LeBeau, J. E., 1803 Bldg. Leng, M. L ., 1803 Loefgren, C., Horgen *CRI, 566 B ldg., Midland CRI, Freeport *CRI, Louisiana CRI, Walnut Creek Tox F ile s , 1803 Bldg. (4) ABCT1s *P, D. Desai, Sarnia R. Montewka, Sao Paulo *R. W. Morgan, 9008 Bldg. *R. D. Moss, Hong Kong R. Rowe, King's Lynn *A. Ruiz, Coral Gables Mattsson, J. L ., 1803 Bldg. M cC ollister, D. 0 ., 1803 Bldg. McDonald, K ., Altona, A ustralia McKeever, D. L ., 2020 Dow Center McKenna, M. J ., 1803 Bldg. McMaster, S. A ., 9008 Nichols, J. L ., Hong Kong N o rris, J. K ., 1803 Bldg. Nowak, R. M., 2040 Bldg. Olson, R. D., Sarnia Pulver, S. M., 2040 Dow Center Quast, J. F ., 1803 Building Rampy, L. W., 1803 Bldg. Rao, K. S., 1803 Bldg. Rausch, D. A ., 2020 Dow Center Reuvers, J. H., Terneuzen Rinzema, L. C ., Horgen Saunders, J. H., 1803 Bldg. Spradling, J. L ., Sao Paulo Steward, D. L ., C incinnati Swank, M. G., 1803 Bldg. T a lc o tt, A. T ., 2030 Bldg. Thcmka, L. M., 2040 Bldg. Venable, J. R., 1803 Bldg. Verschuuren, H. G., Rotterdam Watanabe, P. G., 1803 Bldg. Yocum, R. H ., 566 Bldg. GLOBAL ANALYTICAL DISTRIBUTION LIST Barros, N., Aratu Salvador Blankenship, M. J ., Altona, A u stra lia Branson, D. R., 1803 Bldg. Dalman, D. A ., Tsing Y i, Hong Kong Darby, N., Ft. Saskatchewan Dishburger, H. J ., 9001 Bldg. Eaves, T ., Surrey, England Fike, R. R., Milano Hippie, J. W./H.C. Tung, G ra nville H oldt, E. H., B-1225, Freeport Horton, W. B ., 1510, Plaquemine Jones, E. M., King's Lynn K e ily , H. J . , Merrell-Dow, Cinn. Kroposki, L. M., Z io n s v ille Entire copy c f report. LeValle, R., Buenos A ire s, Argentina Lewis, B. J ., Stade Linowski, J. W., 1776 Bldg. M o ffat, R. W., New Plymouth, NZ Monrabal, B ., ' Tarragona, Spain M ujica, L- Mexico C ity Palacios, F ., B ilbao, Spain P ierzynski, B ., Sao Paulo Ramirez, C., Bogota, Colombia S te h l, R. H., Terneuzen Strasser, J. P ., Gotemba Tokyo Streck, R., Rheinmuenster Vicente, S ., Guaruja Sao Paulo W ilkin s, D., Merrell-Dow, France v 4705 0012847 P ' goi V to a. DIOXIN AND HUMAN HEALTH DOM 2113892 During the past decade, considerable controversy has developed over the human health e ffe cts o f exposure to TCDD, an unintentional contaminant from the production of some herbicides and a n tib a c te ria l agents and a substance created by the process o f combustion. This controversy has been fueled by reports on the e ffe c ts o f Agent Orange exposure on returning Vietnam veterans, the Love Canal s itu a tio n , the explosion of a 2 ,4 ,5-trichlorphe nol plant in Seveso, I t a ly , and, most re c e n tly , the buy-out by the U.S. government of Times Beach, M issouri, a town contaminated with d io x in , ( i . e . , 2, 3, 7, 8-tetrachlorodibenzo-p-dioxin [TCDD]). During th is time period, much speculation and misinformation regarding th is Issue have been generated, w ith re s u ltin g confusion and apprehen sion 1n the public mind. I t 1s the purpose o f th is paper to state the facts regarding TCDD and human health and to place the Issue 1n it s proper perspective. This paper w ill document the follo w in g key points: C ertainly 2,3,7,8-TCDD 1s a potent and to x ic compound th a t can cause a v a rie ty of 111 health e ffe c ts at re la tiv e ly low doses 1n laboratory animals. We should be concerned w ith reducing any unreasonable ris k o f human TCDD exposure in our environ ment. Based on the preponderance o f s c ie n tific stu d ie s, the trace environmental presence o f TCDD does not pose a health hazard to people. There 1s a wealth o f epidemiological data supporting the position th a t humans are fa r less susceptible to TCDD than several of the animal species which have been studied. The mere presence of TCDD in the environment ( s o il , water, fis h , e tc .) 1s not proof o f contamination from in d u s tria l chemical ^sources. The s c ie n tific evidence does not demonstrate th a t TCDD exposure causes cancer 1n humans. There 1s no v a lid epidemiologic evidence Unking TCDD exposure in humans with a higher-than-normal rate o f b ir th defects. 2 ,3 ,7 ,8 -te tra c h lo ro d ib e n z o -p -d io x in 4707 00083CC -la - TABLE OF'CONTENTS What 1s 01ox1n ? ............................................................................................... The American Medical Association Report .. . ...................................... Seveso.................................................................................................................. Epidemiological Studies ................................................................................ Soft Tissue Sarcoma............................................................................................ The Dow M o rta lity Study .................................................................................... B irth D e fe c ts ....................................................................................................... The T o xicity o f TCDO 1n Animal Species......................................................... Environmental Hazard Evaluation (S o il/F is h ) .............................................. References........................................................................................................ 2 4 7 9 10 17 19 21 24 28 DOH 2113893 4708 0008307 -2- What is Dioxin? The term "dio xins" (or more s c ie n tific a lly , "chlorinated d1benzo-pd lo x ln s ") re fers to 75 s im ila r, but d iffe re n t substances.1 Each o f the 75 dioxins th a t has been studied has it s own unique id e n tity and to x ic o lo g ic a l properties. D0H 2113894 On the basis o f animal studies, 2,3,7,8,-tetrachlorodibenzo-p-dioxin (TCDD) is considered to be the most to x ic of the dio xin s, and i t has been comprehensively examined fo r Its to x ic o lo g ic a l e ffe c ts . This report w ill generally address the health impact o f TCDD exposure. i TCOD is a highly to x ic compound th a t can cause a v a rie ty of 111 health e ffe cts at re la tiv e ly low doses 1n laboratory animals. We must be concerned w ith establishing safe levels o f human TCDD exposure 1n our environment. TCDD 1s formed as a trace unwanted contaminant in the production of 2,4,5-trlchlorophenol from tetrachlorobenzene. The herbicide 2,4,5trlchlorophenoxyacetlc acid (2 ,4 ,5 -T ), other trichlorophenoxyaclds, and the germicide hexachlorophene are synthesized from 2,4,5-trlchlorophenol and may contain trace amounts o f TCDD and other polychlorinated d i o x i n s . 2 Dow has not produced the herbicide 2,4,5-T 1n the United States since 1979 due to reduced market demand from an Environmental Protection 4709 0008308 V 4- \* 3 Agency (EPA) emergency suspension which re s tric te d some applications o f the product. There are no manufacturers c u rre n tly producing 2,4,5-T in the United States. A Dow jo in t venture company produces 2,4,5-T in New Zealand. Dow cu rre n tly supplies 2,4,5-T from inventories fo r the rice and rangeland applications presently approved by EPA. The trace amount o f TCDD present in Dow-inventoried 2,4,5-T is less than 10 parts per b illio n TCDD can also be created by combustion. Data Indicate th a t chlorinated dioxins may arise from the combustion o f most types of organic m a te ria l, and suggest th a t chlorinated dioxins re s u lt from trace chemical reac tio n s occurring in f ir e . 3 DOM 2113895 Many chemical reactions occur during combustion at very low concentra tio n s -- parts per b illio n and lower. With more advanced a n a lytica l c a p a b ilitie s , s c ie n tis ts have discovered numerous sources where various dioxins have been formed, Including TCDD. Some o f these sources Include refuse In c in e ra to rs , wood-burning stoves, gasoline- and diesel-powered automobiles and tru cks, fire p la c e s , char coal g r i l l s and c ig a re tte s . TCDD and the other dioxin compounds have been entering the atmosphere via airborne p a rticu la te s from these various sources and s e ttlin g on s o il and 1n bodies of water. There have been a number o f In d u s tria l Incidents throughout the. world where people have been exposed to TCDO. Epidemiological studies have 4710 0008309 DOM 2113896 -4- evaluated these Incidents and a ll of the evidence so fa r tends to reduce the i n i t i a l concern about the health e ffe cts o f TCDD on humans. An ove rall look at the mounting data has been undertaken by the American Medical Association. These findings and conclusions are of In te re s t and bear mention. The American Medical Association Report In 1981 the American Medical Association (AMA) published a technical re port reviewing the medical evidence regarding the to x ic ity and long term health e ffe c ts o f the TCDD contaminant in Agent Orange*. The re p o rt's preface stated: "In s p ite of the voluminous d a ta ...th e re 1s s t i l l very l i t t l e substantive evidence fo r many of the alleged claims th a t have been made against these compounds (d io x in s ). The most serious of these allegations assert th a t Agent Orange, or compounds o f a lik e nature, have caused malignant tumors, spontaneous abor tio n s and b irth defects. Although data from studies on e xp e ri mental animals tend to support some o f these claim s, 1t 1s not ce rta in th a t the animal data are extrapolatable to man. No lab ora tory animal can f u lly s u b s titu te fo r man; we must, th e re fo re , depend on the re su lts of ongoing epidemiological studies on persons who are known to have been exposed." (Agent Orange was a 50/50 herbicide mixture o f 2,4,5-T and 2,4-D pro duced according to sp e cifica tio n s set by the U.S. government fo r use as a d e fo lia n t in Vietnam to protect American troops from enemy ambush.) 4711 0008310 Q -4 < S 6 7 -5- A close look at the summary o f the report gives a clearer in s ig h t Into what the AMA's Advisory Panel on Toxic Substances found. The AMA Panel reported th a t, 1n addition to the marked v a ria tio n s in the s e n s itiv ity and s u s c e p tib ility of animal species to a ll to x ic substances, there are s ig n ific a n t differences between some o f the to x ic e ffe cts of TCDD 1n experimental animals and the human experience. People appear to be less sen sitive to the health e ffe c ts o f TCDD than some o f the animal species studied. DOH 2 1 1 3 8 9 2 One of the more pronounced b io lo g ica l e ffe c ts o f heavy exposure to TCDD, as well as a number o f other chlorinated aromatic compounds, 1s a tendency to cause chloracne in c e rta in animals and humans, the AMA Panel reported. This p a rtic u la r skin condition 1s regarded as the c lin ic a l marker fo r TCDD over-exposure 1n people. The Panel noted th a t hunan systemic d is orders from exposures to TCDD are u n lik e ly to occur 1n the absence of chloracne. Such fin d in g s as Impaired liv e r and kidney fu n c tio n , gastro in te s tin a l I r r i t a t i o n , muscle and nerve disorders, and depression and I r r it a t io n of the central nervous system have been reported a fte r expo sure to r e la tiv e ly large amounts o f TCDD. However, these disorders have not been progressive, and they have disappeared w ith tim e, the AMA report stated. In s h o rt, 1f there is no chloracne, there are u n lik e ly to be other p e rsiste n t to x ic e ffe c ts from exposure to TCOO. The AMA Panel also reported th a t other to x ic e ffe c ts o f TCDD, when experim entally produced 1n te s t animals, appear as pathological changes in the l iv e r , lymphoid and e p ith e lia l tis s u e . ^ 0008311 DON 2113898 -6- TCDD 1s a powerful enzyme Inducer. In addition to a lte rin g normal enzyme a c t iv it y , 1t may potentiate or in h ib it the action o f other to x in s . The AMA report stated th a t TCDD may act as a promoter o f carcinogenesis 1n some stra in s o f rats and mice. For cancer to s ta r t, 1t must go through several d is tin c t steps. The f i r s t stage, called in it ia t io n , is produced by a carcinogen (an I n it ia t o r ) and probably involves irre v e rs ib le mutational changes. Subsequent growth and development of an actual cancer may require promotion, which may be the re s u lt o f additional applications of a carcinogen, or o f other, non-cardnogenic m aterials. Promotion may Involve additional unknown processes which could aid the growth of the cancer or reduce resistance to the developing cancer. TCDD is among the group o f chemicals th a t may secondarily influence the formation o f cancer by promotion. The carcin og enicity associated with TCDD 1n laboratory animals is ty p ic a lly accompanied by other signs o f systemic t o x ic it y 1n contrast to some other chemical carcinogens fo r which cancer 1s the only observable to x ic e ffe c t. TCDD has reportedly Induced genetic changes 1n some forms o f b a c te ria , but evaluations 1n rodents and humans do not Indicate th a t these mutational changes w ill occur 1n higher animals, le t alone people. The AMA Panel made clear in Its summary th a t the extensive Inform ation 4713collected fo r more than 30 years provides no conclusive evidence th a ti 0008312 _p D0H 2113899 -7- 2,4,5-T and/or TCDO are mutagenic, carcinogenic or teratogenic 1n humans, nor th a t they have caused reproductive d if f ic u lt ie s 1n people. Furthermore, the AMA report stated th a t 2,4,5-T undergoes re la tiv e ly rapid decomposition 1n the s o il. However, TCDD does p e rs is t 1n s o il longer than 2,4,5-T, but, 1n general, the half-11fe of TCDO 1n s o il may be no longer than one year. TCDD exposed to u ltr a v io le t lig h t is broken down ra p id ly when present as a th in film on plants, water and the surface of s o il. 4 Seveso The most well-known and widely studied Incident o f community exposure to TCDD followed the July 10, 1976, explosion at the ICMESA trlchlorophenol p la n t at Seveso, Ita ly . As a re s u lt o f th a t accident, approximately seven square miles o f the countryside were contaminated by chemicals, Including TCDD, exposing as many as 30,000 people. 56 The heaviest con tam ination Involved an area o f 180 acres (Zone A), occupied by 736 people. TCDD was reportedly detected on s o il at le v e ls 1n excess o f 5.5 parts per m illio n (ppm).7*5 n general, l i t t l e e ffe c t was noted on pla nt l i f e , but w ild animals did d ie , p a rtic u la rly 1n Zone A (nearest the chemical p la n t). About fou r percent of the domestic animals 1n contaminated zones died, and v ir t u a lly a ll of these were small animals. 5 Some o f these deaths may have been due to other chemicals released at the same tim e. 37- W 0008313 -8- Chloracne, which has generally been regarded as the hallmark or sentinel sign of TCDD exposure8, was observed 1n th is population, although at generally low rates and p rim a rily 1n c h ild re n .9,10,5,6 highest Incidence of chloracne reported was about 13*14 percent among elementary school children in Seveso, and th is was generally mild and ra p id ly resolved 1n most affected people.5.6,11 other than chloracne (o f which there were u ltim a te ly 100*200 cases), nausea, vomiting, itc h in g and head ache were the most commonly observed symptoms8 re a d ily a ttrib u ta b le to the exposure which may have Involved chemicals other than TCOO. Head ache, stomach and in te s tin a l upset were more commonly noted in In d iv id u a ls w ith chloracne. 88 DOM 2113900 B1santil2 concluded th a t no "major event" had taken place with respect to b irth defects, miscarriages (spontaneous abortions), b irth s and deaths. While th is seems to be the consensus, changes in b irth defect reporting and in the re porting o f "spontaneous" (sometimes "thera p e u tic") abortions have obscured any small changes which might have occurred.13,5,6,14,11 Data on reproductive outcome are lim ite d 1n number, but ReggianlH reports th a t examination of fetuses 1n 34 cases o f medically-induced abortion revealed no evidence of In ju ry a ttr ib u ta b le to TCDD. Pregnancy outcome 1n the years follow ing 1976 appears to be comparable w ith Western experience. 6 Of p a rtic u la r In te re s t is the fa c t th a t no b irth defects were observed during 1977 among the 70 b irth s in Zones A and B. Only those pregnan cies coming to term a fte r January 1977 would have been re le va n t, 1.& :'j 4715 0008314 D0H 2113901 -9- those in which July 1976 maternal exposure would have taken place during the c r itic a l period fo r the developing fetus. Many factors are known to cause b irth defects in humans which occur in three to six percent of liv e b irth s : v ira l Infections, genetic predis p o s itio n , smoking, alcohol, and even medications, including excess quantities of certain vitam ins. Epidemiological Studies In recent years a number o f Investigations have been conducted concern ing the human health e ffe c ts associated w ith exposures to TCOD. The follow ing b r ie f review o f the human exposure experience with TCDD and the epidemiological studies o f those people are taken from a recent re p o rt by the Advisory Committee on P esticides, which is charged with making recommendations concerning the safety and safe use o f pesticides in the United Kingdom. Monsanto's plant at N itro , West V irg in ia : 121 workers were followed up fo r 0 years a fte r exposure to TCOD. No apparent excess In to ta l m o rta lity or in deaths from cancer or diseases o f the c irc u la to ry system was observed.15 C oalite plant (1968) in the United Kingdom: Chloracne was I n i t i a l l y seen in 79 workers and some nad i n i t i a l abnormali tie s 1n the U ve r but these returned to normal lim its w ith in 10 days. No c lin ic a lly recognizable disease has been demon s tra te d . There 1s no evidence o f U v e r or cardiovascular d is e a s e .15 0008315 -10- Dow Chemical Company workers exposed to 2,4 ,5 -T : No adverse e ffe c ts have been noted in an epidemiological study in these workers (m o rta lity was a c tu a lly less than expected).15 Vietnam: Exposure to Agent Orange is blamed fo r a host of illn e s s e s . The U.S. Veterans Adm inistration found th a t no cause-effect re la tio n s h ip was established between TCDD exposure and alleged medical com plications.15 Finnish ra ilro a d workers using 2,4,5-T and other phenoxyacetic acid herbicides in fo re s try and on roads and railw ays: 1,960 men were studied ana no increased m o rta lity from cancer was demonstrated. There were no deaths from cancer in fo re s try workers although six deaths would have been s t a t is t ic a lly p re d icte d .15 Soft Tissue Sarcoma D0K 2113902 The most p e rsiste n t hypothesis deals w ith the possible causal associa tio n between TCOO and s o ft tissue sarcomas. This hypothesis 1s derived from the epldenlological work done by Dr. Hardell and his associates in Sweden.16 ' Soft tissu e sarcoma is a generic term fo r a group o f lesions th a t Include more than 100 d iffe re n t types o f tumors. i7 These tumors are r e la tiv e ly ra re ; thus few pathologists have been able to achieve a high degree o f consistency 1n th e ir diagnosis.18 Most experts believe th a t 1 f TCOO were acting as a carcinogen 1t would cause an Increase 1n one type o f s o ft tissu e sarcoma, but would be u n lik e ly to raise the ris k a ll across the board. One o f the reasons v in y l chloride 1s accepted as a human carcinogen 1s the s p e c ific way 1t acts. At high le v e ls , 1t produces a single type o f cancer: angiosarcoma of the l iv e r . i 4717 0008316 DOW 2 I13903 -11- The question which arises is : I f there is a common exposure lin k between various reports of s o ft tissue sarcomas in occupational groups, 1s TCDD the lin k ? The exposure documentation d iffe r s from study to study. In some, based on d e tailed technical knowledge of the chemical process, analyses o f reaction contaminants, and c lin ic a l evidence of excessive group exposure (such as chloracne), there is evidence th a t TCOD is at least one o f the common exposure lin k s . In other re ports, the evidence is more presump t iv e , end 1n some cases i t 1s t o t a lly lacking . Epidemiologists from Dow Chemical and Monsanto have reported on the so ft tissue sarcoma cases occurring in th e ir occupational groups presumably exposed to high levels o f 2,3,7,8-TCDD.1920,21 In 1964, a group o f Dow trichlorophenol production workers experienced an outbreak o f chloracne. Sixty-one workers were id e n tifie d as being involved w ith the trichlorophenol process. Forty-nine o f these workers developed a rash diagnosed as chloracne. Epidemiologists followed up on the status o f a ll 61 men through 1978, 14 years a fte r the in c id e n t. S t a t is t ic a lly , 7.8 deaths would have been expected 1n th is group, but only four had occurred. Among the causes o f death, one was from cardio vascular disease when 3.8 would have been expected. Cancer was Id e n ti fie d as the cause o f three deaths when 1.6 were expected. None o f these fin d in g s represent a s t a t is t ic a lly s ig n ific a n t deviation from the expected experience. 20 One o f the cancer deaths 1n the Dow group was 0008317 -12- due to a type o f s o ft tissue sarcoma called fibrosarcoma. In 1981 another employee 1n the Row trichlorophenol worker group developed another type o f so ft tissue sarcoma, malignant fibrous histiocytom a, one o f a m ultitude o f tumors which comprise the s o ft tissue sarcoma category. The employee died in 1983. In the 1978 Monsanto study, no lin k between exposure to TCDO and excess deaths due to cancer or other causes was found among 121 current and former workers employed at a plant in N1tro, West V irg in ia . The employees who p a rticip a te d in the study Included workers who developed chloracne as a re s u lt of over-exposure to TCDD in a manufacturing accident 1n 1949. Among the Monsanto workers studied, 32 deaths were reported versus 46 which would be expected. Cancer accounted fo r nine deaths among the Monsanto group compared with s lig h tly more than nine deaths expected. One of these cancer deaths was a ttrib u te d to .s o ft tissu e sarcoma, a malignant fibrous histiocytoma. A second Monsanto study evaluated the m o rta lity experience o f male hourly workers employed at the N1tro plant during or a fte r 1955 and followed through 1977.22 when adjustments were made fo r p rio r exposures to a known bladder carcinogen, the observed deaths due to a ll causes and due to cancer were less than expected. Among the subset o f employees who had worked 1n 2,4,5-T production, the proportion o f deaths due to cancer was lower than th a t found in the rest o f the p la n t population. In th is subset one Individual died as a re s u lt o f a s o ft tis s u e sarcoma, a liposarcoma. 0008318 DOW 2113904 -13- These four s o ft tissue sarcoma cases-were summarized 1n the lite r a tu r e 1n 1981.23*24 f our had occupational exposures to trlc h lo ro p h e n o l. Only one, the second Monsanto case, had also been exposed to 2,4,5-T. Three had frank chloracne and the fo u rth , a Dow employee, a fa c ia l rash during h1s assignment 1n the trlchlorophenol p la n t. A ll four had a h is to ry of c ig a re tte smoking. I t was suggested th a t the hypothesis of soft tissue sarcomas being caused by heavy TCOO exposure among c ig a re tte smokers needed to be te ste d . Three other cases o f s o ft tissue sarcoma were also reported 1n 1981.25,26 Exposure to 2,4,5-T, trlchlorophenol or TCOO was not well documented. In one case, the Individual was considered unexposed u n til h1s disease was discovered. The other two occurred 1n a father and son. DOW 2113905 In a group o f Dow 2,4,5-T employees (none o f whom had chloracne)27 there have been no cases o f s o ft tissue sarcoma. At Dow, the m o rta lity experience o f 204 persons exposed to 2,4,5-T during I t s manufacture from 1950 to 1971 was studied through 1978. No adverse e ffe c ts were observed w ith respect to occupational exposure to e ith e r 2,4,5-T or Its feedstock, 2,4,5-trlchlorophenol. M o rtality of the studied workers was favorable compared to th a t o f U.S. white males. There have been no cases o f s o ft tissu e sarcoma reported 1n a BASF worker group, many o f whom experienced severe chloracne. Seventy-four ; 4720 0008319 -14- members o f the BASF group were exposed to TCDO during a 1953 accident in a trichlorophenol production u n it or during cleanup and re p a ir follo w in g the accident. Exposures were heavy. Twenty-seven years a fte r the accident a m o rta lity study o f those exposed to TCOO 1n the accident was undertaken. Overall m o rta lity did not d if f e r in th is group from the ra te expected in three external reference populations or from th a t observed in two internal comparison groups.28 Dow is cooperating with the National In s titu te fo r Occupational Safety and Health (NIOSH) to prepare a study o f more than 4,000 in d u s tria l workers p o te n tia lly exposed to d io xin s. DOM 2113906 Swedish Epidemiological Studies Or. H a rd e ll's work, which has lead to the question about s o ft tissue sarcomas, uses a d iffe re n t methodology than in the foregoing studies -- the case control approach -- and reports on d iffe re n t patterns and leve ls o f exposure.16*29 in his f i r s t study, he reported a f iv e - to s ix -fo ld increased ris k o f so ft tissue sarcomas fo r those presumably exposed to phenoxyacetlc acids, predominantly 2,4,5-T, and chlorophenols ("p re sumably" because the exposure inform ation was s u b je c tiv e ly reported by the study p a rtic ip a n ts and/or next o f k in ). Objective records were incomplete and were d i f f i c u l t to in te rp re t. In the second study, he also estimated the ris k associated w ith exposure to phenoxyacetlc acids not contaminated with.TCDD and found the same magnitude o f r is k . 4721 OOOS3ZO -15- Some epidem iologists reviewing the-inform ation fin d th is confusing. The ju s tific a tio n fo r combining chlorophenols and 2,4,5-T was the presumption o f a common contaminant, TCDD.29 i f th a t were the case, increased ris k would not be expected fo r the phenoxyacetlc acids without th is contaminant. In both of h1s studies, Dr. Harden f i r s t administered mail questionnaires and then followed up w ith telephone interviews on a selected subset o f subjects. While the questionnaire was designed with the idea of masking the in te n t of the in ve s tig a tio n , the telephone inquiry s p e c ific a lly focused on the exposures o f p e rtine nt in te re s t: phenoxyacetic acids and chlorophenols. DOW 2113902 In te re s tin g ly , in his second study, Dr. Harden reported the frequency o f the various exposures among both cases and controls and most of the ris k estimates were elevated. The ris k from only one out o f 15 chemi ca ls, sodium ch lo ra te , was not elevated. . Nicotine had a high ris k . U nfortunately, the ris k estimates derived from the questionnaire alone are not known, nor 1s i t known what the ris k estimates would have been fo r other agents, 1f they had been probed 1n the same depth as were the phenoxyacetlc acids and chlorophenols. There are a number of other concerns re la tin g to H a rd e ll's work. The accuracy w ith which exposure to the chemicals is determined is o f v ita l Importance, and the memory re c a ll techniques Dr, Harden used are subject to e rro r. Workers are u n lik e ly to remember w ith accuracy the chemicals they used some years 1n the past. I t 1s also extremely d i f f i c u l t to 4722 estimate the extent or duration of the exposures. Furthermore, 0 0 8 3 2 1 -16- id e n tific a tio n and c la s s ific a tio n of the various soft tissue sarcomas have not been consistent. The d iv e rs ity of tumors is p a rtic u la rly d i f f i c u l t to explain. I f a ll the tumors had been o f a single type, the data would be much more convincing. Furthermore, research c u rre n tly underway does not support H ard ell's hypothesis concerning s o ft tissue sarcoma: The U.K. Government Advisory Committee on Pesticides noted in February 1983: " I t 1s our view that the procedures used fo r establishin g the Swedish study groups, and some aspects o f the c o lle c tio n o f the exposure data, were not re lia b le ." D0H 2113908 Smith and associates3^ are conducting a case-control study 1n New Zealand. Herbicide ap plication 1s a registered profession in New Zealand, and phenoxyacetlc acids have been used in bulk since the la te 1940s. In a prelim inary report presented at the 1982 In te r national Symposium on Chlorinated Dioxins and Related Compounds, Smith noted he was unable to establish an association between the use of phenoxyacetlc acids and s o ft tissue sarcoma. In fa c t, not one of the s o ft tissue sarcoma patients had ever worked as a licensed phenoxyacetlc acid herbicide a p p lic a to r. Milham31 explored th is Issue 1n the state o f Washington using death c e r tific a te s . While there has been considerable exposure over the years to phenoxyacetlc acids and chlorophenols in Washington s ta te , occupations related to th e ir use did not show consistent patterns o f death due to s o ft tissue sarcomas. S u rp risin g ly, the two occu pations with the highest proportional m o rta lity ra tio s were marine engineers and bankers. Among the deaths in the U.S. A ir Force "Ranch Hand" Study, none were due to s o ft tissu e sarcomas.32 RUhimaki studied 1,960 Finnish herbicide applicators and found no d iffe ren ce 1n death rate from any natural cause, including cancer, compared w ith the to ta l Finnish male population. There were no s o ft tissue sarcomas reported in th is group o f workers.33 The Michigan Department of Public Health (MDPH) is Investigating the possible re la tio n s h ip between dioxin and a small number of s o ft and connective tissue cancer deaths among women in Midland County (where Dow has f a c i l i t i e s ) . 34 In a recent report (May 1983) MDPH 4723 ^ 0008322 DON 2113909 -17- did not Id e n tify a s p e c ific association between any environmental fa c to r and the reported increase of so ft tissue cancer 1n the county. While overall age-adjusted cancer m o rta lity rates in Midland County were below the state of Michigan average, s o ft and connective tissue cancers among white females 1n the county were elevated. As part o f the I n it ia l study, MDPH reviewed data fo r 28 other U.S. counties 1n which TCDD or other dioxins were lik e ly produced as a chemical manufacturing contaminant. They found no Increase o f so ft tissue cancer 1n these counties versus counties th a t did not have In d u s tria l manufacturing sources suspected o f generating such dio xin s. Various animal species did not develop s o ft tissue sarcomas when fed TCOO during laboratory experiments. Additional research 1s under way in the United States and elsewhere. The re su lts o f these studies should o ffe r a be tte r perspective on th is Issue. But rig h t now, the preponderance of evidence does not demon s tra te a lin k between TCOD exposure and s o ft tissue sarcomas. The Dow M o rta lity Study In 1976 a research study was published th a t summarized the m o rta lity experience o f over 8,000 men employed by The Dow Chemical Company. The population of In te re s t was defined based on a March 1, 1954, employee census o f the Midland, Michigan, manufacturing location o f the Company.35 The study o rig in a lly developed background m o rta lity data on the Dow worker population so th a t findings obtained from smaller groups directed at more s p e c ific questions, (e .g . questions regarding p a rtic u la r chemi cal exposures or processes), could be placed 1n perspective. . The observation period fo r m o rta lity follow -up was 1954 through 1972, and 0008323 > -6 D0H 21(3910 -16- comparisons were made to the m o rta lity experience of the corresponding United States white male population. At th a t tim e, m o rta lity among Dow employees was found to compare favorably with th a t o f the general U.S. population. The objectives o f the most recent update of th is group o f 8,181 employees were to extend the observation period through 1978, and to include comparisons with a community-based group of employed men not engaged in chemical manufacturing a c tiv itie s . In a d d itio n , the e ffe cts o f c ig a re tte smoking on m o rta lity were Investigated. The Dow group consisted o f personnel from research departments, the corporate headquarters, and a major manufacturing d iv is io n o f the Company. I t has been estimated th a t as many as 500 d is tin c t chemical processes ranging from small batch operations to large continuous and highly-automated production u n its have been onstream at the Midland manufacturing s ite , Including production o f trlchlorophenol and related p r o d u c ts .36 For the most p a rt, the men 1n the study represent long-term Company employees. Some were hired p rio r to the 1940s. Cause of death was obtained from death c e r tific a te s fo r a ll but nine of the 1,932 deceased members o f the study population. 4725 ' V ' H S 3U 0008324 I I 6 E I I Z HOQ -19- Overall m o rta lity 1n the Oow group was 19 percent less than th a t expected fo r the corresponding United States population. Expressed in another way, a Dow worker o f age 25 1n 1972 could expect to liv e 2.2 years longer than a non-Dow worker o f the same age. There were no s ta tis tic a l findings o f excess m o rta lity among Dow employees fo r any cause of death category. For to ta l malignant neoplasms, there were fiv e percent fewer deaths observed than expected based on the United States white male population. S ig n ific a n tly fewer deaths were observed than were expected fo r a ll categories. Presumably, i f low -level TCDD exposure caused a v a rie ty o f increased illn e s s e s , the opposite from what 1s stated above would have been found. Comparing the Dow workers w ith a comparable employee population in Michigan, the overall m o rta lity ris k was about 10 percent lower in the Dow group than the corresponding non-Dow group.37 B irth Defects Concerns th a t TCDD may cause b irth defects are not substantiated by epidem iological science. A b r ie f review o f some areas of concern and the corresponding re s u lts , and a more s p e c ific look at the Dow s itu a tio n 1n Midland, Michigan, are presented: Aerial spraying o f 2,4,5-T in Oregon: Claims o f a lin k between incidence ot miscarriages and 2,4,5-T herbicide could not be established when a ll the data were avail a b le .15 0008325 D0H 2113912 -20- A six-year study o f a g ric u ltu re and fo re s try workers in Hungary exposed to 2,4,5-T : bata do not suggest tha t there has been any s ig n ific a n t increase of b irth d e fe c ts .*5 New Zealand workers exposed to 2,4 ,5-T : An alleged abnormally high incidence of neural tube defects (such as spina b ifid a ) were chance occurrences, and there is "a very high assurance o f safety in normal use" o f 2,4,5-T.15 Australian worker exposure to 2,4-0 and 2,4 ,5-T : Claims of high incidence of b irth defects were Investigated, and available information revealed no evidence th a t the b irth defects were caused by exposure to these two compounds.15 Facial c le ft defects in Arkansas: Study of exposure to 2,4,f>-Y from 1948-74 reveals these defects to have no association with 2 ,4 ,5 -T .15 About fiv e years ago i t was reported th a t the b irth defect rates 1n Midland County (Michigan) were three times above those fo r the state of Michigan. At th a t tim e, the Michigan Department o f Public Health (MDPH) examined th is issue. I t found an increase had occurred between 1971 and 1974 due to additional reporting o f minor congenital malformations. Before 1971 and a fte r 1974, the b irth defect rates 1n Midland County were e ith e r at or below state rates. I f widespread chemical contamina tio n o f the environment was responsible fo r b irth defects, the rates should have gone up and stayed up. They did not. The changes may have been due to reporting techniques, not additional cases o f b irth defects. I t is very Important to note tha t the numbers of county b irth s and b irth defects were sm all, and thus, rates calculated from them tended to be more variable from year to year than the comparable rates fo r the e n tire s ta te . The MDPH recently announced (May 1983) an updated re port which concludes th a t the b irth defect elevations noted 1n the e a rly 1970s were not unique and th a t no ad ditional studies are warranted a t th is tim e .58 0008326 n o w 21 13913 -21- In 1982 Dow s c ie n tis ts published a report o f a survey of reproductive events among 370 wives o f Dow Michigan D ivision employees exposed to d io x in s .39 They analyzed the associations between exposure and spon taneous abortion (m iscarriage), s t i l l b i r t h s , in fa n t deaths, and several categories of congenital malformations before and a fte r c o n tro llin g fo r m u ltip le confounders ( i . e . , sources o f e rro r or confusion) independently and in various combinations. They did trend analysis to determine i f duration o f paternal exposure was related to adverse pregnancy outcome. There were no associations between any exposure and adverse pregnancy outcomes. These findings have recently been confirmed in another study conducted by A ustralian s c ie n tis ts ^ who examined the impact on b irth rates in vo lvin g Vietnam veterans p o te n tia lly exposed to Agent Orange. The T o x ic ity o f TCDD in Animal Species There are wide va ria tio n s in how d iffe re n t animal species are affected by TCDD. For example, the single oral dose LD50 (the amount th a t 1t would take to k i l l 50 percent o f the te s t animals) 1n guinea pigs is 0. 6- 2.0 ug/kg (micrograms per kilogram or every 2.2 pounds o f body weight [a microgram 1s one m illio n th o f a gram; a kilogram is 1,000 grams, or 2.2 pounds]).41*2 in hamsters, however, TCDD is much less to x ic , w ith an oral LD50 o f 1157-5051 ug/kg.^3,44 Therefore, the guinea pig is approximately 5,000 times more sen sitive than the hanster, although both species are closely related b io lo g i c a lly . The d ire c t relevance o f any single animal experiment to humans must be evaluated w ith respect to a ll other ava ila ble data. 0H 21139H -22- In acute and subchronic studies, liv e r to x ic ity is a prominent component o f TCOO t o x ic ity in ra ts , mice and ra b b its , but not 1n monkeys, where e ffe c ts on the bone marrow and e p ith e lia l tissue are more prominent. Thymic atrophy in ea rly to x ic ity studies suggested th a t TCOO might decrease the immune response. In laboratory animals, s c ie n tis ts can Induce 1mmunotox1c e ffe c ts with high doses of TCOO. However, as the Seveso in cid e n t shows, these e ffe cts have not been demonstrated in exposed humans.45,46 Teratogenicity/Animal Species Of the various to x ic responses o f animals to TCOO, the p o ten tial fo r te ra to g e n ic ity (b ir th defects) has perhaps received the most a tte n tio n . Teratogenic e ffe c ts re s u ltin g from TCOO have been realized only in mice, which show an increased frequency of c le ft palate, along w ith an abnor m a lity o f the central c o lle c tio n system o f the kidney. I t 1s well known th a t many fa cto rs can cause b irth defects 1n mice, including the stress o f being transported by a ir during pregnancy, or being deprived of d rin king water overnight. In both rats and mice, dose le ve ls o f TCDD have been Id e n tifie d at which these e ffe c ts do not occur. In ra ts , TCOO does not cause a teratogenic e ffe c t but s u ffic ie n tly high doses can cause embryo- and fe to to x ic ity . 0008328 -23- Many of the studies w ith TCDD in monkeys have been conducted at the U niversity of Wisconsin. In experiments where the high dose level of about 0.011 ug/kg/day TCDD 1n the d ie t was given to the monkeys fo r up to 9.3 months, there were substantial to x ic e ffe c ts .47,48 A prelim inary abstract of a follow-up study49 showed th a t monkeys given die ts contain ing approximately 0.0017 ug/kg/day o f TCDD had only s lig h t to x ic ity . There are studies cu rre n tly in progress at the U niversity o f Wisconsin at lower doses. D0H 2113915 In a three-generation reproduction study50 o f ra ts at incremental dose levels o f TCDD 1n the d ie t, high doses caused decreased f e r t i l i t y and neonatal s u rv iv a l. The intermediate dose level caused decreased f e r t i l i t y and other e ffe cts in the f i r s t two generations, but not in the th ir d . At the low-dose le v e l, there was no Impairment o f reproductive capacity through the three consecutive generations, indicating that 0.001 ug/kg/day was a no-adverse-effect level over m u ltip le generations. Mutagenicity/Animal Species A number o f mutagenic studies have been conducted w ith TCDD. The major i t y o f the tests have been negative or uninterpre table . Only two o f the te s ts were p o sitive 1n one s tra in of Salmonella b a cteria , showing th a t there is low p o s s ib ility fo r mutagenesis w ith TCDD in b a cte ria . In studies46,51,52,53,54,55,56,57 w ith rats or hunan c e lls , there is l i t t l e in d ica tio n that TCDD e lic it s a mutagenic response. 4730 0008320 -24- Thus, while a few p o s itiv e or questionable mutagenic responses have been observed in ce rta in plant or m icrobial te s t systems, there appear to be no d e fin itiv e correlates o f mutagenicity in higher animals or humans. Carcinogenicity/Animal Species Carcinogenic studies follo w in g ingestion of TCDD have been conducted in ra ts and mice.58,59,60 Review o f these data indicates good co rre la tio n o f the re su lts in rats and mice, with a carcinogenic response associated only with life tim e Ingestion o f higher dose levels th a t also induce other t o x ic it y . The liv e r was the primary target fo r cancer in both ra ts and mice. No cancer response occurred at dose levels o f 0.0010.0014 ug/kg/day in the ra ts and 0.001-0.03 ug/kg/day in mice. DON 2 I I 39 16 TCDD does cause cancer in animals. However, 1t is only a t very high dose le ve ls -- dose levels th a t are higher than those th a t e l i c i t other kinds of t o x ic it y . Cancer Induced by exposure to TCDD would be preceded by substantial signs o f t o x ic it y . This to x ic ity (which 1s reversible when exposure stops) would act as a s e n tin e l, or warning at dose levels below those th a t might cause cancer. Environmental Hazard Evaluation Soil Potential human exposure to TCDD from environmental contamination concerns many people. The follo w in g factors should be considered when 4 7 3 1 evaluating p o te n tia l human health hazards due to TCDD 1n s o il: 0008330 D0H 2113917 -25- TCDO 1s tenaciously bound by s o ils and other m aterials (carbon, charcoal), and studies have shown tha t plants grown 1n s o ils containing TCOD do not accumulate or translocate the m a te ria l. The amount o f material absorbed depends on contact w ith the skin, or ingestion. Soil binding o f TCDD reduces the amount th a t can be absorbed through the skin or in te rn a lly from Ingestion. Animal studies show th a t only a small amount of TCOD 1s absorbed from contaminated s o il. In animal studies, when such s o il con tained about 500 parts per b illio n (ppb) ppb TCDO and when the so il was held 1n contact with skin fo r 24 hours (covered w ith aluminum f o i l ) , about 1/1000 o f the TC00 in s o il was absorbed. Estimates o f absorption from the Seveso Incident suggest th a t people would be expected to absorb about 1/2000 o f the average amount o f m aterial present 1n 1 square meter (approximately 1.2 square yards) of th e ir environment each day. I f one uses the laboratory or Seveso data, one can estimate th a t 100-1000 ppb TCOD 1n s o il could re s u lt 1n human absorption o f as much as a dose equivalent to th a t which produced no e ffe c t when fed to laboratory ra ts fo r a life tim e . I f one assumes th a t.s o il 1s eaten 1n s i g n if i cant amounts by c h ild re n , the permissible level 1s le s s , perhaps 10 ppb. 0008331 DON 2 1 1 3 9 1 8 -26- Based on the assumptions and calculations presented here, (p a rtic u la rly those based on the work of Dr. Kingsley Stevens) , 61 i t appears that general environmental s o il contamination in the range o f low parts per b illio n would not pose a health hazard to the general population, Including the unborn, ch ild re n , pregnant women or the old and In firm . For the general population, excluding children with intim ate s o il con ta c t or those who ingest s o il, a level o f 100 ppb would s t i l l pose no hazard. In occupational s e ttin g s , where workers are protected by standard in d u s tria l hygiene practices, higher leve ls o f contamination would not be accompanied by greater exposure or hazard. Dioxin and Fish Rats consuming a d ie t containing 22 parts per t r i l l i o n (ppt) ppt TCDD (1 nanogram TCDD/kg of body weight per day) fo r a life tim e showed no adverse health e ffe c ts due to TCDD. Rats consume about 10-20 percent of th e ir body weight 1n food each day, while people d a ily consume about 2-3 percent o f th e ir body weight. Thus, fo r a given TCDD concentration, rats w ill Ingest more o f the compound than people. A person eating about 2-3 pounds o f food per day uniform ly contaminated w ith 25 ppt TCDD would Ingest about 0.5 nanogram/kg body weight/day or about one-half the dose (weight o f TCDD per u n it o f body weight) shown to produce no adverse health e ffe c ts when fed to rats fo r a life tim e . Again, th is 1s tru e because people eat less food 1n proportion to th e ir body weight compared to laboratory ra ts . *3 ' D 2-1 0008332 -27- The dose received by eating contaminated fis h is much less. Most people in the United States (99 percent according to U.S. government fig u re s) eat less than 0.6 pound o f fis h per week. Nine out o f ten people in the United States eat less than 0.3 pound o f fis h per week, while the "average" person eats s t i l l le ss. Since fis h is such a small part o f the American d ie t, 99 percent o f the American public would receive less than 1/70 o f the amount o f TCDO shown to produce no e ffe c t in laboratory ra ts fed TCDO d a ily fo r th e ir life tim e s , i f those people ate fis h con ta in in g 25 ppt o f TCDD. Or said another way, a person could eat nearly one ton of fis h per year containing 25 ppt o f TCDD and not exceed the n o -e ffe ct level established by laboratory animal experiments. The fis h consumption guideline o f 25 ppt established by the Food and Drug Adm inistration assures an adequate margin o f safety to protect people from overexposure to TCDD.62 *** D0H 2113919 August/1983 ( 6) 0008333 D0H 2113920 -28- REFERENCES 1. Kociba, R. J. and Schwetz, B. A ., (1982): T o xicity o f 2 ,3 ,7 ,8 -T e tra chlorodibenzo-p-dioxin (TCDD). Drug Metabolism Reviews, 13(3):387-406. 2. G a ra ttln i, S., (1982): TC00 Toxicology with P a rticu la r Reference to Seveso: Introductory Remarks. Drug Metabolism Reviews, 13(3):345-353. 3. Bumb, R. R., Crummett, W. B., et a l . , (1980): Trace Chemistries of F ire : A Source of Chlorinated Dioxins. Science, 210, pp. 385-390. October 24, 1980. 4. B eljan, J. R., MD, et a l . , (1981): The Health Effects of "Agent Orange" and Polychlorinated Dioxin Contaminants, Technical Report, Prepared by the Council o f S c ie n tific A ffa irs ' Advisory Panel on Toxic Substances, Dept, o f Environmental, Public and Occupational Health, American Medical Association, Chicago, IL , October 1, 1981. 5. Homberger, E., et a l . , (1979): The Seveso Accident: Its Nature, Extent and Consequences. Ann. Occup. Hyg., 22:327-370. 6. Pocchiari, F ., et a l . , (1979): Human Health Effects from Accidental Release o f Tetrachlorod1benzo-p-dioxin (TCDD) at Seveso, Ita ly . Ann. N.Y. Acad. S c i., vv:311-320. 7. F a n e lli, R., et a l . , (1982): TCDD Contamination in the Seveso Incident. Drug Metab. Rev. 13(3)407-422. 8 . Crow, K ., (1978): The Chemical Oisease. Hew S c ie n tis t. A pril 13, 1978. 9. Caramaschi, F ., et a l . , (1981): Chloracne Following Environmental Contamination by TCDD in Seveso, Ita ly . International J . Epidemiology, 10(2):135-143. 10. Fara, G. M., et a l . , (1980): Chloracne A fte r Release of TCDD at Seveso, Ita ly . In: Chlorinated Dioxins and Related Compounds: Impact on the Environment, Oxford: Pergamon Press, pp. 545-559. 11. Regg1an1, G., (1979): Estimation o f the TCDD Toxic Potential in the Light of the Seveso Accident. Arch. T o xico l. Suppl., 2:291-302. 12. B ls a n ti, L ., et a l . , (1979): Experience o f the Accident o f Seveso. Proceedings of the 6th European Teratology Society Conference, September 4-7, 1978, Budapest, Hungary, 1979. 13. Abate, L . , et a l . , (1980): M o rta lity and B irth Defects From 1976 to 1979 1n the Population Living 1n the TCDD Polluted Area o f Seveso. In : Chlorinated Dioxins and Related Compounds: Impact on the Environment, pp. 571-587, Pergamon Press, Oxford. 4 4735 0008334 00H 2113921 -29- 14. Regg1an1, G., (1977): Toxic Effects o f TCDD 1n Man. NATO Workshop on Ecotoxlcology, G u ilfo rd , England, July-August, 1977. 15. K ilp a tric k , R., et a l . , (1980): Further Review o f the Safety fo r Use 1n the U.K. o f the Herbicide 2 ,4 ,5-T, Report of the Advisory Committee on Pesticides, M in is try o f A g ric u ltu re , Fisheries and Food, United Kingdom, December, 1980. 16. H ardell, L. and Sandstrom, A ., (1979): Case-Control Study: Soft Tissue Sarcomas and Exposure to Phenoxyacetlc Acids or Chlorophenol. B r itis h Journal o f Cancer, 39:711-717. 17. Hajdu, S. I . , (1981): Soft Tissue Sarcomas: C la s s ific a tio n and Natural H istory. CA-A Cancer Journal fo r C lin ic ia n s , 31(5):271-280. 18. Rubin, P., Ed., and Bakemeler, R. F ., Assoc, e d ., (1978): Soft Tissue Sarcoma, W. B. Patterson, In : C lin ic a l Oncology fo r Medical Students and Physicians. Chapter XIX, American Cancer Society, pp. 210-217. 19. Zack, J. A. and Susklnd, R. 0 ., (1980): The M o rta lity Experience of Workers Exposed to Tetrach1orod1benzod1ox1n 1n a Tr1chlorophenol Process Accident. Journal o f Occupational Medicine, 22:11-20. 20. Cook, R., et a l . , (1980): M o rta lity Experience o f Employees Exposed to 2,3,7,8-Tetrach1orod1benzo-p-d1ox1n (TCDD). Journal o f Occupa tio n a l Medicine, 22:530-32. 21. Zack, J. A ., Gaffey, W., (1980): A M o rta lity Study o f Workers Employed at the Monsanto Chemical Plant 1n N itro , West V irg in ia , Unpublished. 22. Zack, J. A. Gaffey, W. R ., (1983): A M o rta lity Study o f Workers Employed at the Monsanto Company Plant 1n N itro , West V irg in ia . In: Hunan and Environmental Risks o f Dioxin and Related Compounds, R. E. Tucker, Ed.; Plenum P ublishing, pp. 575-591. 23. Cook, R. R ., (1981): D1ox1n, Chloracne, and Soft Tissue Sarcoma. L e tte r to the e d ito r. The Lancet, March 14, 1981, p. 618. 24. Cook, R. R., (1983): Soft Tissue Sarcoma: Clues and Cautions. In : Human and Environmental Risks o f Dioxin and Related Compounds, R. E. Tucker, Ed.; Plenum Publishing, pp. 613-618. 25. Moses, M., and S e llk o ff, E. J . , (1981): Soft Tissue Sarcoma, Phenoxy Herbicides, and Chlorinated Phenols. L e tte r to the e d ito r. The Lancet, June 20, 1981, p. 1370. 26. Johnson, F. E., Kugler, M. A ., and Brown, S. M., (1981): L e tte r to the e d ito r. The Lancet, July 4, 1981, p. 40. 4736 0008335 DON 2113922 I 30. 27. O tt, M. G., et a l . , (1980): A -M o rtallty Analysis o f Employees Engaged in the Manufacture o f 2,4,5-Trichlorophenoxyacetic Acid. Journal o f Occupational Medicine, 22:47-50. 28. Thless, A. M., et a l . , (1982): M o rta lity Study o f Persons Exposed to D1ox1n 1n a Trlchlorophenol-Process Accident That Occurred 1n the BASF on 13 November 1953. American Journal o f In d u s tria l Medicine, 3:179-189. 29. Eriksson, M., et a l . , (1979): Case-Control Study on Malignant Mesenchymal Tumors and Exposure to Chemical Substances. Lakartldnlngen 76:3872-75, (EPA T ranslation). 30. Smith, A. H., et a l . , (1982): The New Zealand Soft Tissue Sarcoma Case-Control Study: Interview Findings Concerning Phenoxyacetic Acid Exposure, Third Intern ation al Symposium on Chlorinated Dioxins and Related Compounds, Salzburg, A u s tria , October 12-14, 1982. 31. Mllham, S ., (1982): Herbicides, Occupation and Cancer. Lancet, 1:1464-1465, 1982. 32. Chesney, M., (1982): Status o f the Ranch Hand Study Concerning Agent Orange, Presentation to the House Committee on Veteran's A ffa irs Subcommittee on Oversight and In ve stiga tions. 33. R11h1maki, V., et a l . , (1982): M o rta lity o f 2,4-Dichlorophenoxya ce tlc Acid and 2,4,5-Trichlorophenoxyacetic Add Herbicide A pplicators 1n Finland. Scand. J . Work. Environ. Health, 8:37-42. 34. "Evaluation o f Soft and Connective Tissue Cancer M o rta lity Rates fo r Midland and Other Selected Michigan Counties Compared N atio nally and Statewide," Michigan Department o f Public Health, May 4, 1983. 35. O tt, M. G., et a l . , (1976): Determinants o f M o rta lity 1n an In d u s tria l Population. J . Occup. Med., 18:171-77. 36. O tt, M. G., et a l . , (1975): Linking In d u s tria l Hygiene and Health Records. Am. Ind. Hyg. Assoc. J . , 36:760-766. 37. O tt, M. G., (1982): E ffects o f Selection and Confounding on M o rta lity in an Occupational Cohort (Doctoral D isse rta tio n ) (M o rta lity Among Men In A Chemical Manufacturing Company -- Executive Summary), U niversity M icrofilm s, Ann Arbor (In Press). 38. "Evaluation o f Congenital Malformation Rates fo r Midland and Other Selected Michigan Counties Compared N ationally and Statewide", 1979-1981, Michigan Department of Public Health, May 4, 1983. 39. Townsend, J. C ., et a l . , (1982): Survey o f Reproductive Events o f Wives o f Employees Exposed to Chlorinated Dioxins. Am. J . Epldea., 115:695-713. DOH 2113923 -31- 40. Armstrong, B ., (1983): A ustra lia Reports No Link Between Service in Vietnam and B irth Defects Among O ffspring. The Epldm. M onitor, 4 ( 3 ):1. 41. Schwetz, B. A ., et a l . , (1973): Environ. Health Perspect. Exp.. Issue No. 5:87. 42. McConnel, E. E., et a l . , (1978): T oxicol. Appl. Pharmacol., 44:335. 43. Olson, J. R., et a l. , (1980): Toxicol. Appl. Pharmacol., 55:67. 44. Henck, J. W., et a l . , (1981): T oxicol. Appl. Pharmacol., 59:405. 45. Homberger, E., et a l . , (1979): The Seveso Accident: Its Nature, Extent and Consequences, Report o f the Glvaudan Research Company. 46. Regg1an1, G., (1980): J . T o xicol. Environ. Health, 6:27. 47. A lle n , J. R ., et a l . , (1977): T o xicol. Appl. Pharmacol., 41:177. 48. A lle n , J. R ., et a l . , (1977): Food Cosmet. T o x ic o l., 15:401. 49. Schantz, S. L ., (1979): T o xico l. Appl. Pharmacol., 48:A180. 50. Murray, F. J . , et a l . , (1979): T o xico l. Appl. Pharmacol., 50:241. 51. Green, S ., Moreland, F. S ., (1975): T o xico l. Appl. Pharmacol., 33:161. 52. Green; S ., et a l . , (1977): DA By-Lines, 6:292. 53. Wassom, J. S ., et a l . , (1977-78): Mut. Res., 47:141. 54. Khera, K. S ., Ruddick, J. A ., (1973): " Polychlorodibenzo-p-d1ox1ns: Perinatal Effects and the Dominant Lethal Test 1n VHstar Rats" , in Chlorodioxlns - O rigin and Fate. Advances 1n Chemistry S eries, No. 120 (Etcyl H. B la ir, e d .), American Chemistry Society, Washington, DC. 55. B eatty, P. W., et a l . , (1975): T o xico l. Appl. Pharmacol., 31:309. 56. Tenchlnl, M. L ., et a l . , (1977): Approaches to Examination o f Gene t i c Damage A fter a Major Hazard 1n Chemical Ind ustry: Prelim inary Cytognie Findings on TCDD-Exposed Subjects A fter Seveso Accident, Special Project o f Investigations on TCDD-Exposed Pregnancies (P ro f. G. B. Candian1 and P rof. L. D eC arll), U niversity o f M ilan, I t a ly . 57. Rehder, H ., et a l . , (1978): Schweiz. Med. Mochenschr., 108:1617. 58. Kodba, R. J . , et a l . , (1978): T oxicol. Appl. Pharmacol., 46:279. 59. National Cancer In s titu te , (1980): DHHS P ublic. No. NIH80-1765. 4738 go8337 -32- 60. Toth, K ., et a l . , (1979): Nature, 278:548. 61. Stevens, K. M., (1981): Agent Orange T o x ic ity : The Q uantitative Perspective. Hunan Toxicology, 1 , pp. 31-39. 62. Cordle, F ., (1981): The Use o f Epidemiology 1n the Regulation o f Dioxins 1n the Food Supply. Regulatory Toxicology and Pharmacology 1, Academic Press, pp. 379-387. 2113924 August/1983 ( 6) 4739 0008338 Io l e  A.'vii. K n\irvn. (V iicim . lo\ici*l. 12. 71 - 76 HW3) 1^ \ " < 2 3 7 2. Q o *j) h n . o/i*f?J 'H r Arcntv*sofEnvironmentalv- Contamination and Toxicology NOT! --z ---ji5 .A^Y 5C C G 'YR'.G'4* t*v- ' wywfiwiw,m m m im i ftBMKWH!WWfcw ID0H2 I 56356 Laundering Procedures for Removal of 2,4-Dichlorophenoxyacetic Acid Ester and Amine Herbicides from Contaminated Fabrics*1 Carol Bryan Easley, Joan M . Laughlin, Roger E. G old, and Duane R. Tupy Department of Textiles. Cluthing. and Design. 234 Home Economies Bldg., University of Nebraska-Lincoln. Lincoln. Nebraska 68583-0802 Abstract. Denim fabrics were contaminated w ith 2.4 - dichlorophenoxyacetic acid (2.4-D) ester and 2,4-D amine form ulations and laundered. The vari ables examined were pesticide form ulations, tem peratures, pre-rinse conditions, detergents, am monia additive, and num ber o f washings. A clean fabric, designated the "tra n s fe r fa b ric ", was laun dered w ith the contam inated denim to assess amounts o f herbicide transferred during laundering. S o lubility o f the form ulation was a m ajor factor in ease o f rem oval and transfer during laundering. The amine is w ater-soluble and was readily rem oved j during the^ laundry process. G enerally. 60C w ater I temperature and heavy-duty liquid detergent w ith} out ammonia additive provided better pesticide reI movai than other tested conditions. Pre-rinse was t not a contributing factor in herbicide rem ovai. but | resulted in s ig n ific a n tly less am ounts o f 2.4-D transferred to concom itant laundry. A second w ashing cycle rem oved s ig n ific a n tly g reater ! amounts o f the herbicide than a single washing. Emphasis on reducing accidental exposure to pes ticides is param ount as pesticide use has increased dram atically in the last fo ur decades (Deichmann 1972). Concern has increased fo r the safe handling _ and usage, environm ental contam ination, and ef fects o f pesticides on non-target species. The three natural routes fo r a compound to enter the human body are oral, respiratory', and.derm al. W olfe e t a t . 11967) sb ou ftd that the m ajor route o f 1Publi->hed as p;ipcrnumber 6*84. Journal Series. Nebraska Ag ricultural Experiment Station. Research was conducted under Protect Nebraska 94-010and North Central Region Pesticide Im pact Assessment Program Project No. 144 pesticide exposure and absorption was through the skin and not the respiratory system. This was ex plained by the fact that only small particles enter the lungs, but larger, more numerous panicles settle on surfaces such as skin, clothing, and soil. Concern fo r dermal pesticide exposure is highest among ag ricu ltu ra l w orkers, farm ers, aerial sprayers, and form uiators because o f the duration o f the applica tio n season. Since the skin is the m ain site o f pes ticide exposure and route o f absorption, attention has focused on clothing items w hich cover and protect skin surfaces. There is evidence that such clothing can lie contaminated by pesticide (Finley at a t. 1969. 1977. and 1979). M etcalfe (1972) found b io lo g ic a lly active m ethyl parathion (.O .O -d im e th yl O -p-nitrophenvl phosphorothioate) (MeP) residue in fabrics after three washings. She also recorded MeP transfer from contaminated fabrics to clean fabrics during concom itant laundering. Recent w ork b y Easley e t a t. (19i> 1) indicated varied success in the rem oval o f MeP from fabrics by laundering. MeP cn uilsifia bie concentrate form ulation was more d ifficu lt to remove from cotton and cotton/ polyester denim fabrics than encapsulated o r wettable pow der form ulations. Fiber content was not a contribu ting facto r in the rem oval o f MeP regardless o f form ulation. O f the laundering procedures inves tigated. pre-rinsing follow ed by a wash and tw o rinses resulted in overall increased levels o f pes ticide removal. Laundry additives o f ammonia o r bleach were relative ly ineffective in assisting the detergent in removal o f MeP. A d ditio na lly. Laugh lin e t a t. (1981) found that washing equipment re tained and provided fo r possible transference to subsequent laundry. The m ajority o f pesticide laundering studies have investigated insecticides: how ever, the largest por- . y. ^ \ C 47 Q 0090-4341/83/0012-0071 S0I.20 J 1983 Sprinser-Verlag New York Inc^ ^ 0 0 0 S t ww C. II. Easley ,/ ,,1. f5E9SIZ MOOt lion o f pesticide ;ictive inurcvlieni (a.i.) applied to crops and/or pusturdnnd in the l ! .S. arc herbicides pended from a drying rack while pipetling was performed, and remained suspended In air dry. (E l*A 19S0. U SD A 1981). C ertain herbicides, in cluding 2.4-1). are under scrutin y by the U .S. E n v i L a u n d e rin g C o n d itio n s ronm ental Protection Agency (E P A ) because o f potential adverse healtlj effects. A p proxim ately 1.500 products containing 2.4-D are registered w ith the EPA. and over 60 m illio n pounds a.i. were used in 1980. This herbicide is used in the c o n tro l o f broadleaf weeds in small grains, field crops, rangeland, and turf. Interest concerning the impact o f 2,4-D on human A contaminated denim swatch was individually laundered along with an uncontaminated transfer swatch in 150 ml detergent so lution calculated proportionally to a standard 45 L washing machine volume. Laundering ofeach switch was done in stain less steel canisters of an Atlas Launder-Ometer (Model B5). A water bath maintained the water temperature fora 12 min deter gent wash and two rinse cycles of five and three min. respec tively. health surfaced follo w in g the E P A ' s emergency suspension o f 2,4.5-T [(2,4,5 -trich lo rop he no xy) acetic acid] and Silvex [2-(2,4,5-trichlorophenoxy) pro pio nic acid] on February 28, 1979. T h is interest was due to the chemical sim ilarities o f 2.4-D and 2,4,5-T and the concern th a t 2,4-D could be con taminated w ith dioxin [2,3,7,8 letrachlorodibenzo- Temperature o f Laundering: Fabrics were laundered in either a 60*C (hot) water wash and 49"C (warm) water rinses or a 3(TC (cold) water wash and 30C water rinses. Although previous studies for launderingcontaminated clothing(Easley et at. 1982) had recommended 60C or above wash temperature, current energy costs have caused lower temperatures for family laun dering (Loveday 1979). />-dioxin (T C D D )] w hich had been linked to fetotoxic and carcinogenic effects. EPA initiated a review to determine if 2.4-D should be investigated under the Pre-Rinse: Fabrics were subjected toeithera "pre-rinse'' or "no pre-rinse" treatment. The pre-rinse was the first step in a fourstep process in which each contaminated swatch was pre-rinsed R ebuttable Presum ption A g ain st R eg istra tio n in 150 ml distilled water for two min priorto laundering. A 49C (RPAR) process. EPA concluded that scientific tox icology studies on 2,4-D did not indicate that con tinued use o f the herbicide posed significant health hazards: however, additional studies were sug gested. water pre-rinse was selected for the 60*C water wash: a 30*C pre-rinse was used with the 30`C water wash. Contaminated fab rics which were not pre-rinsed were laundered in the standard three-step process of a wash and two rinses. Transfer fabrics were not subjected to the pre-rinse cycle: these swatches were introduced at the washing cycle. The purpose o f the present study was to in v e s ti gate laundry procedures required to remove 2,4-D from contaminated fabrics and evaluate the poten tial' fo r dermal exposure fo llo w in g laundering. Detergent: Since previous work had identified heavy duty liquid ( H D D detergentsand high phosphate detergents as effectivefor mu'ny1-araihion removal (Easley et al. 1982). these two deter gents were used. The non-phosphaie H D L was Dynamo- -, a nonionic liquiddetergent manufactured by theColgate-Palmolive Co. The 12% phosphate detergent was American Association of M aterials and Methods Textile Chemists and Colorists (AATCC) Standard Detergent 124. widely used in textiles research. Detergent solutions were fitb r it \: Two types of fabrics were used: lldenim fabrics repre sented pesticide-contaminated clothing laundered in a home laundry situation: and 2) transfer fabrics represented textile items laundered in the same load with eontaminaicd fabrics. An proportionally prepared using 0.2 ml H D L detergent/150 ml water |0.13% solution(manufacturer'srecoinmendaiion>| and 0.3 g A A T C C detergent;!50 ml water (0.2% solution (AATCC stan dard method)|. 80% cotton-20% polyester blend 12M twill denim greige goods! was chosen fur the contaminated fabric. A SOcotion-50% polyester double In, similarin weight toan infant'sT-shirt(5.7 gm-'i. was the selected transfer fabric. The transfer fabric was laundered cuncotnuantly with a contaminated denim fabric. Laundry Additive: Ammonia (NH,) additives are used in the home laundering process to assist detergents in cleaning. Com mercial pesticideapplicators use ammonia-water solutions tode contaminate spray equipment. Therefore, contaminated fabrics were laundered with sudsy household ammonia as a laundry ad Chemicals: Either 2.4-D ester or 2.4-0 amine solution was used to contaminate denim swatches. A 1.25% solution, commonly used foragriculturalapplication, was prepared from field formulalions (amine formulation: alkamil amine sails, 2.4-D acid ditiveor withoutammonia inthe washingsolution. The ammonia (3.5%-4% ammonia concentration) was added on a basis of 0.4 ml/150ml water,equivalent to 250 ml ammonia additive per45 L washing machine volume* equivalent V>.5" (1.7 kjv'3.79 Ll: ester formulation: propylene jdyeol butyl ether estcr^-2.4-0 acid equivalent 44.yi (1.8 kgI 3.70 L)|. Repeated Washing: The efTcctofrepealed washings on pesticide removal had received limited investigation (Metcalfe 1972). Contaminated fabrics were laundered through one complete Fabric Cimtnminawm: Denim swatches. K x 8 cm. were con taminated hv pipettingtwo ml of2.4-D formulation onto the fab cycle ortwo complete laundry cycles inthisinvestigation. Using a 2 x 2 x 2 x 2 * 2 x 2 Woek design (pesticide formulation. ric. A magnetic stirrer provided uniform agitation of the 1.25% i' solution during the contamination process. Swatches were sus *The use of tradenames does not signify product endorsement & 4742 0005720 Rcniov;il of 2.4-D (.'oiiipimnd.s from Contoinmaleil K ih iio ~3*-- n temperature. pre-rinxo. detergent. ;imnu>ni:i additive. repeated washing1.the contaminated denim fabrics were laundered along with uncontaminated transfer fabrics. Unlaundered contami nated fabrics (controls) were analyzed as an indicator of initial contamination and as a baselinettlCf)forcalculating percent pes; ticide removed and percent transferred. All variables tested in this study were replicated three times. E x tra c tio n P ro ce d u re s with a 10-ml aliquot of CHCI,: following separation. CHCI, a W added to (he combined extracts in the round bottom flask. Con9C tents in the flask were concentrated to five ml b> rotary MicuinffO evaporation. The chloroform extract was transferred to a test- tube and further evaporated to 0.10 ml using a N, stream. Fv^jy lowing addition of BF,-inethanol. the procedures were the sang^ as described for2.4-D esterextraction. The 2.4-D amine transfer , fabric extractions did not result in an emulsion formation foj- lowing the addition of CHCI,: therefore, only two 35 ml chlortFJ form extractions were perl'ormed. ,** r Procedure development indicated analysis was more effective in the 2.4-D methyl ester stale. Therefore, techniques were devel oped to convert 2.4-D ester and amine to 2.4-D methyl ester. J.-t-D Ester: 2.4-D ester is essentially insoluble in water, but soluble in most organic solvents. Experimentation with appro priate solvents for extraction was undertaken during procedure development. Maximum recovery was obtained with acetone. Fabric swatches were extracted twice with two-150 ml aliquots of acetone in a two hr mechanical shaking process, as described by Easley et tit. (1981). Aliquots of 2.4-D ester extract were saponified by adding 2 ml of a solution made from 6.4 g potas sium hydroxide (KOH) in 10 ml H.O plus 90 ml ethylene glycol (EC) (Aly and Faust 1963). The resultant solution was evapo rated to2 ml under an N. gas stream which removed the acetone. An additional 8 ml KOH-EC were added. The solution was vor tex stirred and heated in a 70C water bath for 10 min. poured into a 60 ml separatory funnel, and acidified with 4 ml 6 N sul furic acid (H-S04). Salt formed during neutralization and was redissolved by adding 25 ml distilledwater. Two 8-ml aliquots of chloroform (CHCI,) were used to extract the 2,4-D acid. The combined CHCI, extractswere evaporated toapproximately0.10 ml with a N. stream evaporator, after which 2 ml boron trifluoride IBF.) in methanol were added toachieve esterification of the 2.4-f) acid. The solution was healed to 70"C for It)min and quenched in an ice bain. Five ml of a saturated saltsolution and It)ml isu-octane were added and vigorously vortexeu for three min to ett'ec: partitioning of 2.4-D methyl ester into iso-octane. Alter phase separation, a 2-mi aliquotofthe iso-octane layerwas pipetted into a vial fur gas chromatographic analysis. The 2.4-D ester transfer fabric extractions were prepared for analysis in the same manner, with the exception that the entire acetone extract wax concentrated by vacuum rotaryevaporation to approximately live ml prior to sapouification with KUH-EG. 2.4- /> A m int: 2.4-D amine is extremely soluble in water <300 lot) g 11-0) and detectability uf residues after laundering needed to lie thoroughly explored. Amine-contaminated laun dered denim swatches were extracted in two-150 ml aliquots of 0.15 .V sodium hydroxide solution during two hr of mechanical shaking. The entire extract was poured intoa 500 ml separatory funnel and acidified with 10 ml 6 N H-SO, (Buchultz 1981). The 2.4- 0 acid was extracted with a 35 ml aliquotofCHCI,. A severe emulsion problem during phase separation was (educed by the addition uf three ml absolute ethanol. Following two min vigor ous slicking, the phases ere allowed to separate. The chloro form phase and emulsion were drained into a 125 ml separatory funnel. A second chloroform extraction was made and combined with the first using the same procedures. Following phase sep aration in the 125 ml separatory funnel, the chloroform extract was drained intoa 250 ml round bottom flask. Remaining water and emulsion in the 125 ml separatory tunnel were re-extracted G a s C hrom atography Procedures A Hewlett-Packard gas chromatograph (Model 5840A) with a flame ionization detector ivas used for analysis. The separation column was 1.83 m x 3 m m I.D. packed with 3% OV-25 on 100-120 mesh Chromosorb W HP. The identical column tem perature was I85C. and temperature was programmed to in crease ata rateof5*G'min to 1891C. Temperature ofthe inletivas 195*C and the detector was 3 W C . Nitrogen carriergas flow was 1,290 ml/hr. airwas 13.200 ml/hr. and hydrogen was 1.320 ml/hr. External standard quantification was performed usinga standard solution of 2.4-D methyl ester tDow Chemical Co.. 99.98% pur ity) with recalibration every fifth sample. Peak area measure ment was electronically calculated by a dedicated microproces sor.Two injectionswere made foreach sample and averaged per replication. The amount of 2.4-D extracted from unlaundered controls and laundered swatches was expressed in nanograms (ngVcm1. S ta tis tic a l A n a ly sis Differences in the amount of 2.4-D ester or amine (ngem*) be tween controls and laundered swatches were expressed in per centages of 2.4-D removed and transferred, and subjected toarc sin transformation. A fitetorial analysis of variance and a Dun can's multiple range test were used for data analysis. Indication of significance was at the p - .1)5 level. Results Due to the com plexity o f the research design, the findings are discussed separately to r 2.4-D ester and 2,4-D amine form ulations. The treatments o f pre rinse, water temperature, detergent, ammonia ad d itiv e , and repeated washing are presented fo r each form ulation. Furtherm ore, the amount o f herbicide transferred during laundering is discussed fo r each form ulation. 2 ,4 -D E s te r F u rm id a tio n The mean percents o f 2,4-D herbicide rem oved from contaminated fabrics ranged from 28.86% to 44.99%. A m ounts transferred ranged from 1.05% to 2.05% o f the in itia l c o nta m in atio n (Table 1). The 741 74 C. H. finsley et nt. 6 s e e s iZ N o a t Table |. Mean percent 2.4-D ester removed by laundering from eontaminaied denim fabric and transferred to concomitantly laundereJ fabnc Treatments Mean *5*removed Mean transferred1 Pre-rinse Pre-rinse wash 36.52 =12.191 1.48* =0.78 Wash (only) 34.50 = 14.99 1.71* =0.80 Temperature Wl'C wash/49C rinse 44.99* = 11.88 2.14* =0.75 kc^v: 30'C wash'30`C rinse Detergent 26.03" = 7.40 1.05* =0.34 HDL 39.80* = 13.67 1.61* =0.85 -r-? r- A A T C C 124 Repeated wash/addilive Detergent , . 31.22* 32.08* = 12.32 = 10.56 1.58* 1.89* =0.75 =0.89 Detergent + NH, 28.86* = 13.35 2.05* =0.82 ts.- Detergent Detergent + N..H.., 2 washes 40.59* 40.51* = 13.09 = 13.53 1.28* 1.16* =0.48 =0.52 1Mean 7r transferred compares the amount (ng/cm1)of 2,4-D ester on control with amount (ng/cm;)in transfer fabric -Mean followed by the same tetter within treatments are not significantly different at/> .05 level <r;\ 1Standard deviation E- low residue rem oval is attributable to the in so lu b il This indicated that the amount o f 2,4-D transferred ity o f 2,4-D ester in water, yet this insoluble form u was sim ilar regardless o f detergent type. r-- lation apparently was carried to and retained in the transfer fabric even though the transfer fabric was taken through a complete laundry cycle. A m m o n ia A d d it iv e : F or both the contam inated denim and transfer samples, detergent w ashing P re -rin s e : No significant difference was found be tween pre-rinsed samples and non-pre-rinsed sam ples, although rem oval was s lig h tly greater fo r 2.4-D ester contaminated denim fabrics w hich had been pre-rinsed. F or the transfer fabrics, pre-rinse alone was generally more efficient than detergent plus ammonia additive in rem oving residues from the fabrics. The use .of ammonia at the concentra tion tested appeared to in h ib it rem oval o f 2.4-D ester. plus laundry resulted in s ig n ific a n tly less 2,4-D transferred to concom itantly laundered fabric than those that were not pre-rinsed. R e p e a t e d W a s h in g : Repeated washing was sign ifi cantly better than single washing in the removal o f 2,4-D ester from the in itia lly contaminated denim T e m p e ra tu re : A significant difference was found between the hot (M)C wash/49C rinse) and the cold fa bric. Repeated washing also resulted in less 2.4-D ester being sorbed by the transfer fabric. (30'C wash/JO'C rinse) laundry procedures fo r both the contam inated denim and tra n sfe r fa brics. The higher w ater temperatures (Table 1) w ere m ore ef 2 .4 -D A m in e F u rm td a tio n fective in rem oving the ester form ulation from the fabric, but also resulted in increased transfer o f the Laundering procedures removed 99.17% to 99.65% pesticide to concom itantly laundered fabric. This o f 2,4-D amine from the contam inated denim fabric may be due to increased so lu b ility o f the ester fo r (Table 2). These levels o f rem oval were due in part m ulation at higher temperatures. to the extreme so lubility p i the amine form ulation in w ater solutions. Differences found in the removal o f D e te rm e n t: The use o f H D L detergent resulted in the 2,4-D amine form ulation, though statistically significantly better rem oval than that o f A A T C C sign ifican t, were not m eaningful in a practical sense S ta n d a rd D e te rg e n t 124 fo r th e c o n ta m in a te d because o f the nearly com plete elim ination by the denim , regardless o f tem perature or laundry proce laundering procedures tested and lo w to xicity o f dure. H ow ever, the am ount o f 2.4-D ester in the this herbicide to humans. M inute amounts (0.02% to transfer fabric was not significantly different when a 0.26%) o f 2,4-D amine were retained by the transfer comparison was made between the tw o detergents. fabric after concom itant laundering. i-. i 4744 0009742 DOHZI 56360 . t ..... : . '- n ;in::nc removed l>y UiumJcrmz irom contamininoli denim fabrics and transferred to concomitantly laundered Mean CJ removed Mean f transferred' 99.53 99.37* =0.49 =0.75 0.08' 0.21* =0.11 =0.15 99.54* 99.36* =0.55 =0.71 0.17* 0.12* =0.16 =0.16 99.65* 99.24* =0.29 =0.81 0.18' 0.11* =0.14 =0.17 .ashes 99.39** 99.17* 99.63* 99.61* =0.38 =0.49 =0.42 =0.37 0.26' 0.25* 0.02* 0.04* =0.11 =0.19 =0.14 =0.08 mpares the amount (ng/cm2)of 2.4-D amine on control with amount (ng'cm-) in transfer fabric same letter within treatments are not significantly different atp .05 level N-> 'icn .ifican i d ifference in herbicide * '.-tec between the four-step washing r irs.-iaded a pre-rinse cycle and the ee-s':?p tre a tm e n t. H o w e v e r, p re vhr.riy reduced the am ount o f 2.4-D rret! as com pared to the three-step " .vg niilcan t differences were not n the hot (60-'C wash/49C rinse) and : -:0 'C rir.se I laundering proce' - r ' - snim o r the transfer fabrics, .s w ater-so iun lc. it apparently ou from the fabric regardless ' cam d iiio re n c c between H D L " :"e Vi'CC Standard D etergent 124 r,'r r\u ucnim contam inated fa brics. ::d-.r;cs. the d ete rg e nts w ere MaiisiicaJIv compared. These "Parsons were sim ilar to those o f the " P!i:;.iion ( f.ih le I). i-indings from the 2.4-D amine transfer fabrics indicated that '^ e may have inhibited residue re? Detergent alone was generally 'rt||s than detergent plus am m onia ad- the d itfc ie n c e was not sig n ifica n t. *' A second washing significantly Ptoval o f 2.4-D am ine from the co n- fabric and decreased amounts o f residue transferred to concom itantly laundered fab rics. Conclusions General conclusions and recommendations result ing fro m this study include: 1) 2,4-D fo rm u la tio n s w hich are water-soluble are more effe ctively re moved through the'laundering process than insoluble fo rm u la tio n s: 2) 6C wash/49" C rinse w a te r temperatures resuit in better residue removal than 30C tem peratures fo r the ester fo rm u la tio n : 3) heavy duty liquid detergent provides fo r greater re moval o f 2.4-D during laundry than does a high phosphate detergent: 4) 2.4-D can be transferred from contaminated clothing to textile items laun dered in the same wash load: therefore, it is im perative that contam inated clothing be laundered separately: 5) pre-rinsing is helpful when laundering contam inated fabrics, and does result in sig n ifi ca ntly less 2.4-D being transferred to other tabrics in the same wash: 6) am m onia as a laundry a d d itiv e offered no advantage in the rem oval o f 2.4- D resi dues during laundering: and 7) tw o launderings o f contaminated fabrics result in greater rem oval than one laundering: however, the effects o f more than tw o washings needs fu rth e r investigation. Differences in the amount o f 2,4-D herbicide re moved from contaminated denim fabrics and trans ferred to other fabrics during concom itant launder ing is dependent upon pesticide form ulation. Solu b ility o f the form ulation was a m ajor facto r in ease o f rem oval through laundering. Because o f the rela- 4745y 0009743 UUH2I 56361 c 'le % i i 3VSii A liv e ly low to x ic ity o f th is herbicide, transfer in lau nd ering is not a m a jo r concern. It could be anticipated that exposure due to prolonged dermal contact could be m inim ized through proper laun dering o f clothing contaminated w ith 2,4-0. References Aly. Osman M.. and Samuel D. Faust: Determination of 2,4dichlorophenoxy acetic acid in surface waters. J. Amer. Water Works Assoc. 55. 639 (1963). Bucholtz. D.: Purdue University. West Lafayette. IN. Personal communication. March (1931). Deichmann. W. B.: Health hazards in farming and gardening. I ed. Chicago. IL.: American Medical Association (1972). Easley. C. B.. J. M. Laughiin. R. E. Cold, and D. R. Tupy: Methyl parathion removal from denim fabrics by selected laundry procedures. Bull. Environ. Contam. Toxicol. 27, 101 119311. Easley. C. B.. J. M. Laughiin, R. E. Gold, and K. L. Schmidt: Detergents and water temperature as factors in methyl parathion removal from denim fabrics. Bull. Environ. Con tam. Toxicol. 28, 239 (1982). Finley. E. L., J. M. Bellon, J. B. Craves, and K. L. Koone: Pesticidecontamination ofclothingincotton fields. La. Agr. 20. 8 (1977). Finley. E. L.. J. B. Craves, and F. W. Hewitt: Reduction of methyl parathion residues on clothing by delayed field re- entry and laundering. Bull. Environ. Contam. Toxicol. 22. 598(I`I79i. Finley, E. L.. and J. R. B. Rogollio: DDT and methyl parathion residues found in cotton and cotton-polyester fabrics worn in cotton fields. Bull. Environ. Contam. Toxicol. 4, 343 (1969). Laughiin. J. M.. C. B. Easley. R. E. Cold, and D. R. Tupy: Methyl parathion transferfrom contaminated fabrics tosub sequent laundry and to laundry equipment. Bull. Environ. Contam. Toxicol. 27. 518 (1981). Loveday. R. M. T.: Textile consumers' laundry maintenance habits, practices and problems in Lincoln-Lancaster County. Nebraska. Master's Thesis. University of Nebras ka, Lincoln. N E 11979). Metcalfe. G- 1.:The absorption and retention ofselected chlori nated hydrocarbons and organic phosphate residues in cot ton and cotton polyesterfabrics. Master'sThesis. Louisiana State University. Baton Rouge. LA (1972). U.S. Department o f Agriculture: Implications of pesticide regu lations. Econ. Research Scrv. Staff Report No. AGESS810730. Washington. DC (1981). U S . Environmental Protection Agency: Pesticide industry sales and usage 1980 market estimate. Office Pesticide Programs. Washington. DC (1980). Wolfe, H. R.. W. F. Durham, and J. F. Armstrong: Exposure of workers to pesticides. Arch. Environ. Health. 14, 622 (1967). Manuscript received March IS. 1982: accepted June 28, 1982. ( 0009744 a^ i o '7 9- H t MNP7 2 2 8 3 Health* Environmental Information For Further Information Contact: R. W. (Bob). Charlton (517) 636-9303 DIOXIN AND HUMAN HEALTH Executive Summary The issue o f dioxin in the environment and the possible human health e ffe c ts o f th is compound has been the subject o f much speculation over the past decade. This paper discusses s c ie n tific research and makes the follow ing major points: The word "d io x in " refers to a fam ily of 75 compounds o f which the one called TCDD* is the most to x ic based on animal te s ts . Dioxins can be produced as an unwanted contaminant o f some manufacturing processes and through some combustion sources. An American Medical Association review o f available research has concluded th a t there is no convincing support fo r allegations th a t TCDD causes cancer, b irth defects or other reproductive d i f f i c u l tie s in humans. Studies o f some 30,000 people exposed to TCDD a fte r an in d u s tria l accident 1n Seveso, I t a ly , found a chemically-Induced acne th a t occurred in approximately 200 people and no p e rsiste n t e ffe cts which could be v e r ifie d . There was no excess o f b irth defects, abortions or deaths in the population. A number o f studies o f workers a ccid e n ta lly exposed to large amounts of TCDD as long as 30 years ago have Id e n tifie d cases of chloracne in some o f those workers. However, no long-term health e ffe c ts have been demonstrated among th is worker group. The studies have looked fo r cancer, heart disease, liv e r damage and other ailments. IN co The work o f a Swedish s c ie n tis t who purports to fin d a lin k between TCDD exposure and s o ft tissue sarcoma (a re la tiv e ly rare cancer) is not consistent w ith other studies. This research is not supported by the bulk o f studies o f exposed in d u s tria l employees, herbicide applicators and others which show no excess o f th is type of cancer. * A study o f 8,000 Dow employees 1n Midland, Michigan, where herbi cides were produced, found th e ir death rate to be 19 percent less than the general U.S. population. There was fiv e percent less cancer than predicted. *2.3,7,8-tetrachlorodibenzo-p-diox1n 474 $008304 1 THE DOW CHEMICAL COMPANY MIDLAND, MICHIGAN 49640 T ) ^ ijg,tf if DOM 2113891 Investigations o f herbicide a p plicato rs, forest workers and Indus t r i a l workers and th e ir wives found no lin k between TCDD exposure and b ir th defects. Studies o f animals exposed to TCDD show a v a rie ty o f adverse health e ffe c ts th a t vary widely from species to species. In most studies, safe le ve ls of dioxin exposure have been Id e n tifie d . Cancer occurred only at leve ls where animals were su ffe rin g from obvious TCDD poisoning. A review o f the s c ie n tific lite r a tu r e suggests th a t humans are less susceptible to TCDD than various animal species. As 1s the case w ith other to x in s , TCDD exposure should be c a re fu lly con tr o lle d . The s c ie n tific evidence to date does not warrant undue concern about in d ivid u a l exposure to trace amounts of TCDD presently measured 1n the environment. Government environmental guidelines c u rre n tly provide fo r more than adequate margins o f safety. 1^^0^008305 S' J 8 4750 CP-^fc, Am. Ind. Hyt- Assoc. J.45(I):S6-62(1984) F /L ^- -Q ) / 9 ^ y p< / Occupational Exposure of Herbicide Applicators to Herbicides Used Along Electric Power Transmission Line Right-of-Way STEVEN L1B1CH*. J A M E S C. T O \ R I C H A R D F R A N K * and G E O R G E J. SIRONS* ASalety Services Department. Health and Safety Division. Ontario Hydro. Pickering. Ontario. Canada: "Provincial Pesticide Residue Testing Laboratory. Ontario Ministry of Agriculture and Food. Guelph. Ontario. Canada - Occupational exposures to herbicides were measured among 12 applicators in 1979 and 24 applicators in 1980, who were applying the three " herbicides, 2 ,4-D, dichloroprop and pidoram to electric power transmission rights of ways. In 1979 only urine was analyzed but in 1980 both ' breathing-zone air samples and urine were analyzed for herbicide residues. Dermal absorption was found to be the major absorption route being up to 50 times greater than exposure by t t ^ nhalation route when using a hand gun sprayer. Even with the mist blower herbicide application method, dermal absorption was 4 anu . times greater than exposure by the inhalation route. Worker education on hazards of _ skin contact and improved protective equipment significantly reduced the 1980 urine concentrations of herbicide residues. A model is presented to relate the urinary concentrations to equivalent daily exposure levels. 1D0H2I 579A6 Introduction Public concern about the potential adverse health effects o f 2 ,4*0 has intensified since the emergency suspension o f 2.4, 5*T and Brushkill mixtures (2.4-D/2.4.5-T) in several prov inces in Canada, including Nova Scotia, O ntario (1979) and Saskatchewan during the late seventies. This concern stems p rim a rily fro m the chemical sim ila rity o f 2,4-D and 2. 4,5-T and the question o f 2,4-D being contaminated by 2.3, 7,8-tetrachlorodibenzo-p-dioxin (2J.7.8-TC D D ). a manu facturing contaminant found in 2.4.5-T. Evidence indicates th a t the harm ful effects o f 2,4,5-T were caused by the T C D D c o n ta m in a n t/1' Recently it has been reported that 2,4-D form ulations contain no 2.3,7,8-TC D D to a level o f 1 ppb (detection lim it 1 ppb), but other dioxins were fo u n d /2' These were 2.7 and 2.8 dichlorodibenzo-pdioxinand 1.3.6,8-tetrachlorodibenzo-p-dioxins. Their con centration ranged fro m 20 ppb to 4200 ppb. however, none has been shown to be a hazard to human health. The objective o f this study was to systematically evaluate the worker's exposure to herbicides during applications to control woody growth along power line corridors. The applicators are exposed to 2,4-D and other herbicides in varying degrees dependent on weather conditions, height o f brush and type o f terrain in which spray operations are conducted. Uptake o f herbicide can be through inhaling the aerosols o r through skin absorption by touching wet ted brush, hose o r spray equipment. This report presents the significance o f such exposures from the occupational health viewpoint. Methods / Structure of Study This studyrfonsisted o f a p re lim ina ry assessment in Ju ly August 1979.8y measuring herbicide concentrations in urine ol workers. A t the time a 2.4-D. picloram m ixture was being ap plied to tall vegetation under trans-provincial power lines on t**4 St the Canadian shield. Improved handling procedures, improved equipm ent, use o f protective clothing and an instru ction al program have taken place since the 1979 assessment. The m ain study was conducted in 1980. T his study consisted o f measuring the exposure routes by taking breathing-air and urine samples during the a pp lica tion o f herbicides at five selected w ork locations. Sampling for Exposure Assessment Air Sampling A n a ir sampling schedule was designed to cover a ll the m ajor types o f spray equipm ent, types o f terrain encountered and types o f herbicides used by the Power U tility 's forestry crews durin g the 1980 summer stem fo lia r a pplication season (Table 1). Breathing a ir samples were taken by adsotption o f the herbicides on ` F luorisil'sorption tubes. A ir from the breathing zone was d raw n through these sorption tubes at the rate o f 200 m L /m in by battery operated personal sampling pumps. Each member o f the spray crew was sampled fo r 5 consecu tive days d urin g the spray operations which averaged 3 hours per day. Urine Sampling The strategy in urine sampling was to obtain (1) a preoperation sample fo r baseline. (2) several weekly samples taken on Thursdays before or after the a ir sampling week, and (3) daily samples during the week o f a ir m onitoring. Each urine sample consisted o f an A M and PM sample which were later combined to form a daily sample. These were then refrigerated and sent to the P rovincial Pesticide Residue Testing Laboratory for analysis. Types of Spray Equipment The field equipm ent used to apply the herbicides consisted o f single or double 1J M I. tanks m ounted on cither a pick-up tru ck'o r an a ll tc tra in \ chicle (A T V ). A pump 4m In H ,i Astec J. ( t i l , 4751 iinuify. 1984 h a v c V g i'-en T L V ; however, because o f its chemical sim ilar ity to 2 .4 -0 , we have used the same exposure guideline. Urine There are no guidelines fo r maximum urine concentrations fo r the chlorophenoxy herbicides. The literature shows that urine concentrations w ill reflect total body burdens o f these chemicals: that is, the total o f inhalation, skin absorption and ingestion. We chose to use urinary guides which would reflect an equivalent dose as would be received by inhalation alone when personnel are exposed to T LV concentrations. T o derive these levels the fo llo w in g urinary model is utilized. Urinary M odel ' Previous studies using human volunteers shi - d that 2,4-D -was absorbed into the blood plasma by a lu st order rate process w ith a half life o f 11.7 hours.*3,41The herbicide was elim inated essentially unchanged into the urine, also by an apparent firs t order rate process w ith a h a lf life o f 17.7 hours. However, data fo r absorption rates o f 2,4-D through the skin are not available. Lavy*51reported that 2,4,5-T which is sim ilar to 2,4-D can be absorbed through the skin and eliminated in the urine by firs t ord er kinetics w ith a h a lf life o f 18 and 23 hours, respectively. In developing this model the absorption rate through skin fo r 2,4-D is estimated to proceed w ith a h a lf life o f 12 hours, which is sim ilar to that o f absorption through the gastrointestinal tract and in the same ratio as the absorp tio n and elim ination rates fo r 2,4.5-T. These considerations provide the basis fo r the model below, w hich shows k. k. D (t) - B(t) - Eft) where D (t) = dose o r am ount o f 2,4-D rem aining on the skin to be absorbed at time t B(t) = am ount o f 2,4-D in the body at tim e (t) E ft) = am ount o f 2,4-D excreted at tim e t k . and k* are first order rate constants fo r absorption and excretion respectively. The differential equations and in itia l conditions describ ing this model can be expressed as follow s The absorption rate constant k . = 1.39 d a y '1 computed from the absorption h a lf life o f 12 hours. S im ila rly the elim ination rate constant k . = 0.94 d a y '1 computed from the elim ination h a lf life o f 17.7 hours. S ubstituting these rate constants the equation representing the elim ination o f 2,4-D herbicide becomes Eft) = D. ( I 2. le '1-'** - 3. le****) Using this equation the plots in Figure I were obtained. From Figure 1the fo llo w in g observations can be made. The m axim um am ount o f 2,4-D is elim inated in the urine d u r in g day 2 fo llo w in g a skin contact w ith this chemical o f dose D as shown by the single dose curve in Figure 1. In 4 days 93% o f the 2,4-D passes th rough the body. I f successive equal doses are applied d a ily then the body burden w ill build up u n til the elim ination rate equals the dose rate as shown by the m ultiple dose curve in Figure 1. This occurs after 4-3 days. The to ta l body burden becomes approxim ately 1.2 times the d aily exposure. Thus, by mea suring the urine concentration on Thursday o r Friday o f a w ork week, the daily exposure can be approxim ated. U rinary 2.4-D concentrations can be transformed to equiv alent daily exposures to airborne 2,4-D. The transform ation ^ - 7 ^ = - k . Dft) - D (0) - D . dt (1) k. D ft)-k.B ft) dt B (0)s O (2) 8 )B/SIZMOOI ~ k .B ft) dt E f0)= 0 <3) 4 The integrated form fo r the cumulative q u a n tity o f 2.4-D elim inated in the urine after a skin exposure is 1.1 k. eu ) / -Er n- " - c n r * k. - ka SI Tin-Myi Figure 1 -- Calculated fraction of applied dose of 2 .4 -0 on the skin eliminated in the urine during each day. Curve A - a single dose applied at tim e zero: Curve B five equal doses applied at beginning of each day followed by zero dose on day^j g g 4 7 5 24m me H rt 4uoc J. 145) ^ Jjnwr*. 1914 P- .. . P 1D0H2157950 Figure 2 -- Weekly average urinary 2.4-D variation for 1979 and 1980. these actions the mean residues o f 2,4-D and p id o ra m were m arkedly reduced in 1980 (Table III). Im proved awareness and education o f the w orker was as much a fa cto r as was the im proved personal protection o f the program . Urinary Mode! Support H The urinary model fo r 2.4-D elim ination predicts that the body burdens o f 2 ,4 -0 should be reduced d u rin g the week end and then increased during the w ork week due to succes sive d aily exposures. The amount eliminated in the urine should show a low on M onday and an increase during the week to a m axim um on Friday. Figure 2 shows the average urine concentrations variation through the week fo r the 1979 and 1980 data. Taking in to account the possible daily variations in individual doses the data show the increase through the w ork week predicted by the m odel. This gives support to its use in predicting equivalent airborne exposure levels fro m the urinary concentrations and enabling a com parison with the exposure guidelines. Figure 2 also shows the difference between 1979 and 1980 average concentration levels in urine and reflects the degree o f improvement in reducing exposure that was achieved by the educational program and modifications in procedures. Exposure Pathway The study conducted in 1980 was intended to determine the importance o f the two m ajor exposure pathways fo r 2,4-D namely the inhalation route and the dermal route, fo llo w in g the in tro d u c tio n o f the new procedures. F o r the inhalation route, herbicide concentrations were measured in the breath ing zone o f the applicators and these appear in Table IV . It was assumed th a t a ll herbicide inhaled was absorbed by the body. Derm al exposure was determined ind ire ctly by calcu latin g the difference between the amount excreted in the urine and that inhaled into the lungs. The results o f urinary analysis appear in Tables 111 and V. Table H I presents the comparative u rina ry residue levels that were taken during the m onitoring week when inhaled a ir was being measured. In Table V the u rin a ry residue levels are presented fo r the pre-spray periods and the period before a n d /o r after the m onitored week to determine i f workers were fo llo w in g the prescribed procedure on a regular basis. The sampling program covered the m ajor p art o f the sea son's spraying program . It was concluded th a t, by and large, applicators were comfortable w ith the new procedures and continued to use them through the season. The amounts o f herbicide inhaled by the workers were based on the fo llo w in g facts and assumptions. Spray a p p li- Herbicide 2.4-0 TABLE IV Breathing Zone A ir Samples During the Spray Operation a t Five Locations During the Week of Monitoring, Ontario 1 9 8 0 Spray Operation (Locations) Number of Number of Herbicide in Air Samples (iig/m J) Applicators Samples Mean S.O. Minimum Maximum Gun - Roadside (4) Gun - R.O.W." (3| Mist Blower R.O.W. (1) 13 9 3 53 (+20)* 38 9 7.1 13.5 55.2 4.9 7.6 30.7 1.0 0.4 16.2 19.5 35.3 91.3 - Oichloroprop Pidoram Gun - Roadside (2) Gun R.O.W. (2) 4 Gun * Roadside (2) Gun-R.O.W. (1) Mist Blower - R.O.W. (1) "Twenty duplicate samples "R O W. rights-of-way. 6 6 7 3 3 CO 27 C20) 26 25 12 9 9.2 5.4 17.2 10.9 1.0 0.5 18.9 44.1 1.3 2.2 <0.2 10.5 2.3 1.1 0.9 5.0 14.9 8.9 1.9 270 4753 0012881 Am !r.i A:tsc 3 5; 14 y TABLE VII Exposure to 2 .4 -0 Among Four Workers at Kapuskasing 1 9 8 0 Where Job Rarely Rotated Between Applicators and Drivers Content in Urine (mg/kg) 2.4-0 Picloram Job Mean S.D. Mean S.D. Applicator 1 2 7.29 2.57 9.11 2.27 0.30 0.15 0.46 0.09 Driver 1 0.64 2 1.07 0.96 1.10 0.13 . 0 20 0.07 * * show th a t the mist blower increased airborne exposure to 2,4-D fo ur fold, and to picloram six fold. While the urinary excretion o f the tw o herbicides increased so did the p ro po r tio n derived from inhalation. W ith 2,4-D the inhalation exposure rose from 3.4 to 9.3% by changing from gun to mist blower. W ith picloram the increase was from 14.3 to 23.5%. Effect o f Herbicides The active ingredients in the 2.4-D / picloram m ixture were in the ratio o f 4: l . Quantities o f these two herbicides in sampled a ir was in the ratio o f 5 to 6:1. In urine the excretionary ratios o f these two herbicides were 20 to 25:1 (Table V I). The active ingredients in the 2 .4 -D /dich lorop ro p m ix tures were in the ra tio o f 1:1. Q uantities o f these tw o herbi cides in sampled a ir were also in this same 1:1ra tio . In urine the excretionary ratios o f the two were in the ratio o f 1:1 o r 1:2. In the case o f the 2,4-D /piclo ra m m ixture the active ingredients were form ulated as amines while the 2 ,4 -D /d ichloroprop ingredients were form ulated as esters. These differences did not appear to affect either dermal absorption o r elimination in the urine. Discussion Psthwey of Entry Several studies have shown that the chlorophenoxy herbi cides such as 2,4-D and 2,4,5-T are absorbed by the body and alm ost 100% o f the absorbed dose is elim inated unchanged via the urine. Thus the u rina ry concentration is a valuable indicator o f the to ta l body burden o f 2,4-D , i.e . that which is absorbed by inhalation, through ingestion and also through the skin. In a previous study o f forestry workers using 2,4,5-T, it was shown that inhalation accounted fo r a very small frac tio n o f the 2,4,5-T entering the body.<s,The m ajor fraction o f the 2,4,5-T was absorbed through the skin. Lavy et reported that with 2,4,5-T exposures, dermal absorption w a s estimated at 1000 times that o f 2.4,5-T in inhaled air.(S) In o u r * study this ratio o f 2,4-D varied from 50 to 11:1 fo r 2,4-D. ^ D uring the handling and m ixing o f the c o n c e n tra te d ^ chemical a very small skin exposure can be equivalent to a ^ f u ll day o f field exposure. A proper assessment o f the p o te n -c o tia l hazard to workers durin g herbicide applications sen dependent on reliable estimates o f the to ta l quantity herbicide absorbed under all conditions. ~* Since 2,4-D is essentially unchanged by the body the u rina ry concentrations can be used as a guide to the m in im um estimate o f the 2,4-D absorbed by the body and to the degree o f exposure as derived in the urinary model. Worker Education The emphasis on w orker awareness and the procedure and equipment changes before the 1980 spray season concentrated on reducing dermal exposure by workers. The 1980 m o n ito r ing results showed that this was the correct emphasis and resulted in a marked decrease in exposure as reflected in the lower urine concentrations. Since dermal absorption is still the m ajor exposure route, the recommendation to the field per sonnel was to further emphasize the value o f protective cloth ing and the importance o f personal hygiene practices in reducing exposure to herbicides. Other factors such as type o f terrain, type o f herbicide and type o f equipm ent were less im p o rta n t in determ ining the degree o f exposure. The only other factor which influenced exposure was the more concen trated herbicide m ix used in the mist blow er application. References 1. Poland. A .P ., A. Smith, G. Malter and P. Posaack: A HealthSurvey of Workers in a 2 .4 -0 and 2.4,5-T Plant with Special Attention to Chloracne. Arch. Environ. Health. 22:316- 327 (1971). 2. Cochrane. W .P.. J. Singh, W. Miles, B. Wakefield and J. Scott: Analysis of Technical and Formulated Products of 2.4-0ichlorophenoxy Acetic Acid and the Presence of Chlori nated Oibenzo-p-dioxins. In Proc. Workshop. Impact of Chlori nated Dioxins and Related Compounds on the Environment. Oct. 22-24. 1980. Rome Pergamon Press (1981). 3. Sauerhoff. M .W ., W.H. Braun, G.E. Blau end P.J. Gehring: The Rate of 2.4-D Following Oral Administration to Man. Toxi cology 5:3-11 (1977). 4. Gehring, P.J. and J.E. Betso: Phenoxy Acids. Effects and Fate in Mammals. Ecol. Bull (Stockholm) 27:122-133 (1978). 5. Lavy.T.L.: Measurement of 2.4,5-TExposure ofForest Works. Project Completion Report. Univ. of Arkansas. Little Rock, AR. 6. Lavy, T.L.. J.S . Shephard and J .D . Mattice: Exposure Mea surements of Application Spraying 2.4.5-T in the Forest. J. Agric. FoodChem. 28:626-630(1980). 22 July 1982: Revised II August 1983 t 0012883 4754 (2 Am InC. HfC Artec J. (AS) Jin-.nr. 193-1 er S) LO IO Or* DOM2 1585D2 \G I= N G R A N G E A N I U M A N I-IIE A I.T I-I -es&b-<5 . r r T r For further information contact: R. W. (Bob) Chartton (517) 636-9303 Dow April 1984 II6BS12AM DIOXIN, AGENT ORANGE, AND HUMAN HEALTH Su-- ary Agent Orange, the United States m ilita r y code name fo r a 50/50 mixture o f the herbicides 2,4,5-T and 2,4-D, was used during the Vietnam War from 1965-1970 to d e fo lia te jun gle vegetation to protect U.S. servicemen from enemy ambush. Both of these herbicides had been used dom estically without noteworthy adverse e ffe c t fo r about 20 years at the time they were selected by the m ilita r y as the components of Agent Orange. Present at trace leve ls In the 2,4,5-T component of Agent Orange 1s an unwanted contaminant, the d io x in compound 2 ,3 ,7 ,8 -te tra c h lo ro d ibenzo-para-dioxin, comnonly known as TCDD or d io x in . TCDD is extremely to x ic and has been shown to cause a number of serious health conditions 1n laboratory animals, Including b ir th defects, cancer, and death. I t is not a product, and no one makes i t on purpose. TCDD has also been shown to cause a serious skin disorder known as chloracne and re versible signs o f tox1c1y 1n workers a c c id e n ta lly exposed to extremely high leve ls on the jo b . The Dow Chemical Company 2020 Dow Center Midland, Ml 48640 fr9SSrZMQQ -n - In spite o f the acknowledged to x ic ity o f TCDD, the consensus of scien t i f i c opinion is tha t i t has not been shown to cause harm in people at the trace levels at which i t has been present in herbicides. Among the organizations which share th is view are the American Medical Association; the United Kingdom's M in is try of A g ric u ltu re , Fisheries, and Foods; the World Health Organization of the United Nations and the Council fo r A g ricu ltu ra l Science and Technology. During the Vietnam War, The Dow Chemical Company supplied about 32 percent of the Agent Orange applied. As a q u a lity control measure, Dow analyzed the 2,4,5-T in a ll shipments of Agent Orange to the govern ment to ensure the absence -- no detectable leve l -- o f TCDD. Today, w ith b e tte r a n a lytica l techniques, we know th a t the level of TCDD in Dow-supplied Agent Orange was less than 0.5 parts per m illio n . In early 1979, the f i r s t of m u ltip le lawsuits was file d against seven manufacturers of Agent Orange: Dow, the Monsanto Company, Diamond Shamrock Corporation, Uni royal In c ., T.H. A g ric u ltu ra l and N u tritio n Company, Thompson Chemical, and Hercules Inc. These claims have been consolidated In to a class action s u it in which thousands o f Vietnam veterans and th e ir fam ily members w ill be represented by selected cases. These cases w ill be trie d In d iv id u a lly to determine whether Agent Orange has caused health problems to these veterans and th e ir fa m ilie s . The t r i a l is scheduled to begin in early May 1984 1n the courtroom of Chief Judge Jack B. Weinstein o f the Federal D is tr ic t Court fo r the Eastern D is t r ic t of New York (Brooklyn, NY). 4*?58 0013436 'V -H D Z i S6S8S I Z M M -m- We a ll appreciate th a t Vietnam veterans have been through tough, demand ing duty, a d i f f i c u l t experience. Although prelim inary find ings, such as the Ranch Hand studies and the Veterans' A dm inistration's Agent Orange R egistry, have been reassuring, s c ie n tific research at th is time is in s u ffic ie n t to determine d e fin itiv e ly whether the health of the Vietnam veteran and his fam ily is any d iffe re n t from the general U.S. population. We do know that overwhelming s c ie n tific evidence demon strates tha t Agent Orange is neither a lik e ly nor plausible cause of the health e ffe c ts some Vietnam veterans and th e ir fam ilies have been s u ffe rin g . Studies which have examined th is issue and found no lin k between Agent Orange and i l l health include the Ranch Hand m o rta lity and m orbidity studies, the A ustralian b irth defect study released in 1983, and the Veterans' Adm inistration Agent Orange R egistry. There are presently about 95 studies ongoing to evaluate the potential health impacts o f TCDD. E s s e n tia lly , only one o f these studies, presently being conducted by the U.S. Centers fo r Disease C ontrol, considers health e ffe cts in the Vietnam veteran from any potential cause other than Agent Orange. 4 4759 0013437 4760  > 0 \ a / 2 l ' & Z ' g ' O L NUMBER GAP 0013438 'D - m i DIOXIN, AGENT ORANGE AND HUHAN HEALTH MS8SI ZMOf l Approximately 2 .ft m illio n servicemen served in Southeast Asia during the Vietnam War, according to the Veterans' A dm inistration. Today, some of these veterans are experiencing a wide v a rie ty of health problems. The Centers fo r Disease Control (CDC) is attempting to determine whether the Vietnam experience is related to these health concerns. One aspect of the Vietnam experience being explored by the CDC, and the one which has received the greatest public and s c ie n tific a tte n tio n , is the claim th a t the i l l health of these veterans is related to Agent Orange exposure. Agent Orange was a d e fo lia n t used by the U.S. m ilita r y from 1965 to 1970 to elim inate dense jungle vegetation and protect a llie d troops from enemy ambush. This d e fo lia n t was a 50/50 mix of 2,4,5-T* and 2,4-D , two herbicides developed as a re s u lt of m ilita r y research during World War I I . Both of these herbicides had been used dom estically, and w ithout noteworthy adverse e ffe c t, fo r about 20 years at the time they were selected by the m ilita r y as the components of Agent Orange. Code named by the orange Id e n tific a tio n band painted on the storage drum, Agent Orange was usually sprayed from fixed wing C-123 m ilita r y a i r c r a f t . A small amount of the herbicide was also applied from ground sources, such as boats, trucks and backpack sprayers. The source o f today's public controversy is an unwanted trace contaminant, the dio xin compound 2,3 ,7,8-tetrachlorodibenzo-para-dioxin (TCDD) > 4762 Note: Underlined terms are defined in the glossary at the back of tM s paper. 0013439 T J -fS S S ' (09SSI2MQq (4 -2- present at trace levels in the 2,4,5-T component of Agent Orange. I t should be noted that TCDD is not a commercial product, but rather an unavoidable manufacturing process contaminant. I t is not shipped in drums or ra ilc a rs across the United States. Considerable controversy surrounds TCDO even today, p rim a rily because of it s high acute t o x ic it y . TCDD has been shown to cause a number of serious conditions in laboratory animals, including b irth defects, cancer and death. However, according to numerous independent s c ie n tific reviews, with the exception of the skin disorder chloracne and rever sib le signs of t o x ic ity , none of these health conditions have been documented in people, even in cases of severe overexposure. Cl Cl 2,3,7,8-TCDD We must be concerned with establishin g safe levels fo r human exposure to TCDD in our environment. However, there is extensive s c ie n tific know ledge about TCDD which does not suggest th a t the trace le ve ls o f th is 0013410  I S8SI ZMDG -3- contaninant found in herbicides or in the general environment pose a healtiy'threat to people. We must also be concerned about the health problems of in d ivid u a l veterans and th e ir fa m ilie s . These veterans have endured d i f f i c u l t , demanding duty in the service o f th e ir country. They deserve answers to th e ir very real concerns. Although prelim inary findings should be encouraging to veterans, s c ie n tific research is in s u ffic ie n t at th is time to determine d e fin itiv e ly whether the health of these veterans and th a t of th e ir fam ilies is any d iffe re n t from the general population. However, a great deal of public anxiety and misunderstanding has developed about alleged lin k s between health problems and Agent Orange. According to overwhelming s c ie n tific evidence, Agent Orange is not a plausible cause of the various illn e sse s experienced by some veterans. The' bulk of the s c ie n tific studies conducted in the United States, Europe, A u s tra lia , and New Zealand among in d u s tria l workers, herbicide sprayers, and m ilita r y personnel demonstrates th a t the presence o f TCDD at trace levels has not shown any long-term adverse health e ffe c ts in people. * 0013411 -4- INDEPENDENT SCIENTIFIC REVIEWS O The American Medical Association Review cn CO cn In 1981, the American Medical Association (AMA) published a technical report reviewing the medical evidence regarding the to x ic ity and long-term health e ffe c ts of the TCDD contaminant 1n Agent Orange. As the re p o rt's preface states: "In sp ite of the voluminous data . . . there is s t i l l very l i t t l e substantive evidence fo r many of the alleged claims th a t have been made against these compounds (Agent Orange, 2,4,5-T , and TCDD). The most serious o f these allegations assert th a t Agent Orange, o r compounds of a lik e nature, have caused malignant tumors, spontaneous abortions, and b ir th defects. Although data from studies on experimental animals tend to support some of these claims, 1t 1s not c e rta in th a t the animal data are extrapolatable to man. No laboratory animal can f u lly s u b s titu te fo r man; we must, th e re fo re , depend on the re su lts of ongoing epidemiological studies on persons who have been exposed."* Some o f what we know o f the potential health and environmental e ffe c ts of TCDD exposure has been due to s c ie n tific studies conducted on laboratory animals. Yet as the AMA Advisory Panel on Toxic Substances which produced the re port concluded, "there are s ig n ific a n t differences between some o f the to x ic e ffe cts o f TCDD in experimental animals and the human experience; thus the animal data cannot be tra n sla te d d ire c tly to man. "2 00134:2 1IS8SIZMOQ -5- In terms o f acute to x ic ity , a wealth of epidemiological data support the position th a t people seem less susceptible to TCOD than several of the animal species which have been studied. According to the AMA Panel, one of the more pronounced b io lo g ica l e ffe cts of heavy exposure to TCDD, as well as a number o f other chlorinated compounds, is chloracne which occurs in humans and some animals. This p a rtic u la r skin condition 1s regarded as the c lin ic a l in d ic a to r o f TCDD overexposure in people. The AMA Panel noted tha t human systemic disorders from exposures to TCDD are u n lik e ly to occur in the absence o f chloracne. Such findings as impaired liv e r and kidney fu n ctio n , g a s tro in te s tin a l i r r i t a t i o n , muscle and nerve disorders, depression and fa tig u e , and central nervous system e ffe cts have been reported a fte r exposure to re la tiv e ly large amounts o f TCDD. However, these disorders " have not been progressive" and have diminished w ith time a fte r exposure ceased, the AMA report stated. In s h o rt, i f there is no chloracne, other pe rsisten t to x ic e ffe c ts from exposure to TCDD are "u n lik e ly ".* The AMA report also stated th a t TCDD may act as a promoter of carcinogenesis In some s tra in s o f rats and mice. For cancer to s ta r t, 1t must go through several d is tin c t steps. The f i r s t stage, called in it ia t io n , is produced by a carcinogen (an I n it ia t o r ) and probably involves irre v e rs ib le mutational (genetic) changes. Later growth and development of an actual cancer may require promotion, which could be 0013453 4766 -fi- the re su lt of additional applications of a carcinogen, or of other noncarcinogenic m aterials. Promotion may involve additional unknown processes which could aid the growth of the cancer or reduce resistance to the developing cancer Q>^ By way of an analogy, the process of carcinogenesis could be compared to the events leading up to the rupture o f a dam. In itia tio n could be compared to an event which produces a s lig h t crack (genetic damage) in the dam, which in and of i t s e l f does not s ig n ific a n tly jeopardize the dam's s tru c tu re . Promotion could be compared to an unusually v io le n t storm which raises the water level in the r iv e r , increasing the water pressure on the crack causing increased flow , and widening of the crack to the point at which the stru ctu re becomes undermined. Im p lic it in th is analogy is the concept of a threshhold in terms o f promotion: tha t is , there is a water level below which the w ell-being of the stru ctu re is maintained, even though a small leak ( in it ia t io n ) was present. i TCDD is among the group o f chemicals th a t may secondarily influence the formation of cancer in animals by promotion. The ca rcin og enicity associated w ith TCDD in laboratory animals is ty p ic a lly accompanied by other signs o f systemic t o x ic it y , 1n contrast to some other carcinogens fo r which cancer is the only observable to x ic e ffe c t. In terms of m utagenicity, TCOO has reportedly been studied 1n various te s ts using certain b a c te ria l, v i r a l , yeast and tissue c e ll c u ltu re s . 4767 V -W 3 0013414 I MHZ I 5851V -7 - While these in v itr o te sts have given mixed re s u lts , tests using in ta c t animals or human c e lls in c u ltu re indicate l i t t l e potential fo r mutagenic e ffe cts from TCDD to occur in liv in g higher animals. The AMA Panel made clear in it s summary tha t the extensive inform ation collected fo r more than 30 years provides " no conclusive evidence" that e ith e r TCDD or 2,4,5-T are "mutagenic, carcinogenic, or teratogenic in man, nor th a t they have caused reproductive d if f ic u lt ie s in the human."3 An updated report from AMA is expected in la te 1984. Other Independent S c ie n tific Reviews Other independent s c ie n tific groups have reviewed the studies on TCDD. United Klngdoa P esticide Advisory Committee Report: According to th is 1980 re p o rt, updated 1n 1983, and commissioned by the Minis t r y of A g ric u ltu re , F ish eries, and Foods, "human health ris k s posed by d io xin contamination in 2,4,5-T formulations may h ith e rto have been overestim ated." The report states th a t it s authors "cast fa r and wide" 1n search o f evidence th a t 2,4,5-T herbicide form ulations, used p ro p e rly, posed a th re a t to e ith e r humans or the environment; however, no such evidence was found. This report c o n s id e r^ the presence of the TCDD contamination in those 0013415 - 4768 -8- herbicide form ulations, recommended s t r ic t control over the contaminant, but found "no va lid medical or s c ie n tific evidence th a t 2,4,5-T herbicides harm humans, animals, or the environment i f they are used in the recommended way and fo r the recommended purposes. Ol i Queensland Cabinet Committee Report: "No evidence exists tha t the continuation of present and approved use o f 2,4-D and 2,4,5-T w ill in any way harm the health and well being o f any members of the general p u b lic ," according to th is 1981 A ustralian re p o rt. Regard ing Agent Orange, the report also states th a t no evidence exists th a t use o f the d e fo lia n t "caused any physical d is a b ility of the local population" and fu rth e r adds th a t the m ilita r y use of these two herbicides provided no in d ic a tio n "th a t th e ir approved use in peacetime would cause any hazard to l i f e ."5 Council for Agricultural Science and Technology Report: On the presence o f TCDD In Agent Orange, th is re p o rt states th a t, "One is a t f i r s t lik e ly to be so overwhelmed by the enormity o f the to x i c ity o f the compound th a t he f a ils to comprehend the InfinitesmaT le ve ls o f exposure." This e a rly , 1975 re port did not examine the Issue o f ca rcin og enicity but found "no conclusive evidence of association between exposure to herbicides and b irth defects In humans." * 00134+6 i 4769 9IS8SIZM0D -9- INDUSTRIAL EXPOSURES TO TCDD From the la te 1940s to recent years, a number of chemical process upsets anti in d u s tria l incidents have occurred around the globe u n in te n tio n a lly exposing people to high leve ls of TCDD. While these incidents are unfortunate, the knowledge gained by studying the exposed populations is important in evaluating the p o ten tial fo r TCDD to cause human harm. In a d d itio n , such knowledge should provide understanding needed to prevent future harm. The Seveso Incident The best known and most widely studied incident of comnunity exposure to TCDD followed the July 10, 1976, process accident at the ICMESA t r i chlorophenol plant at Seveso, I t a ly . As a re s u lt of the accidental release, approximately seven square miles of the countryside were contaminated by chemicals, including TCDD, exposing as many as 30,000 people.^8 The heaviest contamination involved an area of 180 acres (Zone A ), occupied by 736 people. TCDD was reportedly detected on s o il at levels in excess of 5.5 parts per m illio n (ppm).9? This concentration is 5,000 times the level of concern la te r set fo r TCDD by the Centers fo r Disease Control (CDC) fo r evaluating the hazard of re s id e n tia l s o il at Times Beach, M issouri. (See Appendix D fo r chart comparing various reported concentrations of TCDD.) I t 1s also p o te n ti- * a lly tens of thousands of times greater than any TCDD exposure ground troops could have had to TCDD from the spraying of Agent Orange in Vietnam. -<C, 0013447 -10- " S P S liK P Q At Seveso minimal e ffe c t was noted on plant l i f e as a re su lt of th is high exposure; but some' w ild animals especially herbivorous species did die , p a rtic u la rly in Zone A (nearest the chemical p la n t). About four percent of the domestic animals in contaminated zones died. (V irtu a lly a ll were small animals.)^ Some of these deaths may have been due to other chemicals released at the same tim e. Chloracne, which has generally been regarded as the hallmark, or sentinel sig n , of TCDD e x p o s u re ,w a s observed in the Seveso population although a t generally low rates and p rim a rily in c h ild re n .^ 12,7 ,8 highest incidence of reported chloracne was about 13 to 14 percent among elemen ta ry school children in Seveso, and th is was generally mild and rapidly resolved in most affected people.^8,13 Q ^e r than chloracne (o f which there were u ltim a te ly 100 to 200 cases), tra n s ie n t nausea, vom iting, itc h in g , and headache were the most commonly observed symptoms re a d ily a ttrib u ta b le to the exposure, which again may have Involved chemicals other than TCDD. Headache, stomach and In te s tin a l upset were more com monly noted in in d ivid u a ls w ith c h l o r a c n e . I n a d d itio n , very s lig h t peripheral nerve impairment was reported, and there were also te n ta tiv e signs of liv e r to x ic ity , both of which reversed over tim e. Immunoresponse was not diminished, nor was s u s c e p tib ility to in fe ctio u s diseases in cre a se d .^ Two years a fte r th is In cid e n t, 12 researchers involved in the study of * reproductive e ffe cts at Seveso concluded th a t no "major event" had 0013448 4771 IIS8SIZMOfl -11- occurred with respect to b irth defects, miscarriages (spontaneous abor tio n s ) , b irth s and deaths.*5 However, changes in b irth defect re port ing procedures and in the reporting of spontaneous abortions may have obscured any small changes which might have occurred.16,7,8,17,11 oata on reproductive outcomes are lim ite d , but one researcher reported that examination of fetuses in 34 cases of medically-induced abortion revealed no evidence of fe ta l in ju ry a ttrib u ta b le to TCDD.* Pregnancy outcome in the years follo w in g 1976 appears to be comparable w ith the Western experience. I t is of p a rtic u la r in te re s t th a t no b irth defects were observed during 1977 among the 70 b irth s in Zones A and B. Only those pregnancies coming to term a fte r January 1977 would have been relevant, i . e . , those in which J u ly 1976 maternal exposure would have occurred during the c r it ic a l period fo r the developing fe tu s . I t should also be noted tha t many factors are known to cause human b irth defects, which occur in three to s ix percent of liv e b irth s : fo r example v ira l in fe c tio n s , genetic p re d isp o sitio n , smoking, alcohol, and even medications, includ ing excess q u antitie s of ce rta in vitam ins. A long-term epidemiological survey of the 220,000 residents o f the Seveso area is being conducted by the National In s titu te o f Environ mental Health Sciences (NIEHS) and the International Agency fo r Research on Cancer (IARC) working group. 00134 9 -12- Other Exposures From In d u s tria l Incidents While the Seveso incident provides considerable in s ig h t as to the repro ductive e ffe cts and acute to x ic ity of TCOD, the latency period is in s u f fic ie n t to draw re lia b le conclusions about c a rc in o g e n ic ity . Studies of other in d u s tria l Incidents are more revealing 1n terms of the potential o f TCDD to cause cancer in people. N1tro, West Virginia: In 1949, an in d u s tria l accident occurred at a Monsanto trlchlorophenol plant re s u ltin g in 122 cases of ch lo racne among the workers. Of those exposed 121 were monitored fo r 30 years a fte r th e ir high peak exposure to TCDO. To date, there is no apparent excess 1n terms o f to ta l deaths, deaths from cancer or deaths from diseases of the c irc u la to ry system. *8 o f the to ta l * cancers, there was one case o f s o ft tis s u e sarcoma. A recently reported 1979 cross-sectional study o f herbicide production workers at th is plant concluded th a t i t is " u n lik e ly " th a t " permanent, severe, and d e b ilita tin g " to x ic e ffe c ts are "in e v ita b le " a fte r exposure to TCDD s u ffic ie n t to produce chloracne. The study also noted th a t Individua l s u s c e p tib ility may make c e rta in heavily exposed workers more vulnerable but found tha t "even severe acute to x ic o lo g ic a l e ffe c ts o f TCDD were re ve rsib le or markedly improved over tim e ."19 * Midland, Michigan: In 1964, a process change in a Dow tr lc h lo r o phenol plant resulted in the unintentional exposure o f 61 workers i 4773 G013450 ais^suNOfl -13- to TCDD levels p o te n tia lly as high as 6,000 to 10,000 parts per m illio n . Of those exposed, 49 workers had some evidence o f ch lo racne. A ll 61 of these workers have been monitored over the 20 years since th e ir exposure. When la s t reported in 1980, no excess had occurred in terms of to ta l deaths or deaths from diseases o f the c irc u la to ry system. (There were, in fa c t, fewer deaths in both categories than s t a t is t ic a lly expected.) Cancer deaths have been s lig h tly elevated (3 deaths v. 1.6 expected), but no single type of tumor was predominant.20 o f the three cancer deaths, there was one case of s o ft tissue sarcoma. An additional study including both Dow and Monsanto occupationally exposed groups is being conducted by the National In s titu te of Occupational Health and Safety (NI0SH). Other exposures to TCDD from in d u s tria l incidents have occurred outside the United States. Bolsover, United Kingdom: As a re s u lt o f a 1968 explosion 1n a C oalite and Chemical Products, L td . plant manufacturing 2,4,5-T from trlc h lo ro p h e n o l, 79 workers developed chloracne, which was the only consistent fin d in g . 21 i n i t i a l find ings on these men suggested some change 1n white blood c e ll counts, and 1n three 0013451 4774 <1 ) ^ 7 0 H S IS U M Q Q -14- cases, an elevation of glucose in the urine, suggesting possible liv e r and kidney damage. But a repeat screening 10 days la te r did not confirm th is pattern: a ll of the tests gave values w ithin normal lim it s . 21,22 should be noted tha t the design of th is study has received strong c ritic is m , since less than h a lf of the workers who developed chloracne were included.23 no fin d in g s on m o rta lity are reported. While most cases o f chloracne were resolved w ith in a matter of months follow ing the accident, new cases were noted in a report nine years la te r which found that one-half the workers studied who had once had chloracne were s t i l l su ffe rin g from "minor" cases of the skin disorder. Other than the chloracne, the study noted " no evidence" th a t the workers had been "adversely affected in any way."24 Ludwigshafen, Germany: Due to a process accident in 1953 at a * BASF A ktiengesellschaft trichlorophenol p la n t, 55 workers developed chloracne.25 peripheral nerve damage and liv e r to x ic ity were reported.26 dU6 to in s u ffic ie n t decontamination, an addi tio n a l case o f chloracne developed fiv e years la t e r .27 Follow-up studies 27 years la te r reported no excess in to ta l deaths, but did fin d an excess of to ta l cancer deaths (7 v. 4.1 expected). Of these, three deaths were due to stomach cancer (v . 0.6 expected).27 However, the study's authors note th a t, "Many of the workers in the facto ry fo r some time had probably been exposed to other chemicals and the exposure to dioxin was an addition al experience o f temporary d u ra tio n ."27 0013432 BON215852* -15- Amsterdam, The Netherlands: As a re s u lt of an explosion at an NV P hilips h e rbicide-fa cto ry in 1963, 145 people were exposed to TCDD, 69 of whom had d e fin ite signs of chloracne.28 Liver function te sts did not ind ica te damage.26 Twenty years la te r , "no s ig n ific a n t diffe ren ces" were found between those workers and a matched control group.28 Data on cancers did not show any organ-related pattern and are not considered to represent any excess m o rta lity . Times Beach and Im p e ria l, M issouri: Mismanagement of waste in 1971 by a now defunct company which produced hexachlorophene led to the spraying o f TCDD-contaminated waste o ils fo r dust control on southern Missouri roads and two horse arenas. Analysis 20 years la te r found th a t the waste o ils may have contained as much as 350 parts per m illio n of TCDD.29 in both o f the arenas, a number o f animals died; and in one o f them, 1n which la te r analysis revealed about 33 parts per m illio n o f TCDD 1n the s o il, reports in d ica te th a t two children developed lesions resembling chloracne.29 (See Appendix D fo r chart comparing various reported concentrations o f TCDD.) Testing in Times Beach and Imperial 1n 1983 revealed TCDD 1n s o il ranging from non-detectable levels to 300 parts per b i l l i o n , a hundred times less than th a t found 1n the horse arena.30 Recent te s tin g o f 68 residents with presumed high exposure to TCDD found no cases of chloracne and no d iffe re n c e , 1n comparison with a control group, 1n b irth defects, I n f e r t i l i t y , Impotence, headaches, loss o f memory or cancer ra te s .31,32 s lig h t ly more urinary 0013433 , "4776 fim iJH Q O -16- tra c t problems and enlarged liv e rs were found in the te s t group, but the differences were not s t a t is t ic a lly s ig n ific a n t. Continued health surveillance of the Times Beach residents is planned. Other In d u s tria l Exposures: In addition to these accidental indus t r i a l exposures, a number o f occupational exposures have occurred at levels high enough to produce adverse e ffe c ts . Limited knowledge 1s available on occupational exposures at 2,4,5-T plants 1n Spolana, Czechoslovakia, from 1965 to 1968; and 1n New Jersey from 1963 to 1969. In both o f these cases, porphyria cutanea ta rd a , a liv e r dysfunction re s u ltin g 1n lesion s, was reported in addition to chloracne.333* These find ings are obviously in con f l i c t w ith those at Seveso and also w ith the s ix other In d u s tria l incidents previously reported 1n th is review 1n which porphyria cutanea tarda symptoms were not Id e n tifie d . As pointed out by the authors o f the New Jersey study, the exposures there probably involved chemicals other than TCDD. A lim ita tio n o f the preceding studies 1s th a t they were conducted a fte r TCDD exposures s u ffic ie n tly high to cause obvious signs o f Illn e s s . A Dow Chemical report on herbicide workers w ith in one of I t s chemical complexes provides Inform ation on the re s u lts o f lesser exposures. This report examined 204 employees w ith p o te n tia l TCDD exposure from 1950 to 1971, and found no chloracne and no other observable adverse * e ffe c ts .3 5 E ffo rts w r t made to minimize TCDD contamination o f the 0013434 4777 -p-3 sesuH O fl -17- product at th is f a c ilit y re s u ltin g in an in te rn a l standard developed in 1965 o f less than one part per m illio n , which was undetectable by a n a ly tic a l standards at the tim e. I t is important to note th a t most of these in d u s tria l incidents repre sent e s s e n tia lly worst-case scenarios involving exposures considerably higher than any th a t could be derived by ground troops, even 1f d ire c tly sprayed w ith Agent Orange: in some cases, the exposures were p o te n ti a lly from 350,000 to ten m illio n times greater. The only la s tin g , consistent fin d in g documented in these in d u s tria l incidents is chloracne. HYPOTHESES REGARDING THE HEALTH EFFECTS OF TCDO Soft Tissue Sarcoma Perhaps the most publicized hypothesis about TCDD and human health Involves the possible causal re la tio n s h ip between the compound and s o ft tissu e sarcomas. In some studies based on detailed technical knowledge o f the chemical process and c lin ic a l evidence of excessive exposure (chloracne) there 1s evidence th a t TCDD 1s at le a st one o f a number of common exposures. In other reports the evidence is more presumptive, and 1n some cases 1t 1s t o t a lly la c k in g . To date, the bulk of the evidence does not demonstrate a causal lin k between TCDD and s o ft tis s u e sarcomas. 0013455 4778 ^ S(S83ISHOO -18- Soft tissue sarcoma is a generic term fo r a group of lesions that include more than 100 d iffe r e n t types of rare cancers.3 Most experts believe that i f TCDD were acting as a carcinogen i t would cause an increase in one type of s o ft tissue sarcoma rather than causing an increased ris k across the board. For example, one o f the reasons vinyl chloride is recognized as a human carcinogen is the s p e c ific way i t acts. At high doses i t has been associated with a single type of soft tissu e sarcoma: angiosarcoma of the liv e r . The s o ft tissue sarcoma hypothesis derives from the epidemiological work reported in 1979 by Dr. Lennart Hardell and his associates in Sweden.37 no one has been able to re p lic a te D r. H a rd e ll's fin d in g s , and his studies have been strongly c ritic iz e d because o f the potential fo r observer and re ca ll bias in addition to other confounding e le m e n ts .38,39 D r. Hardell f i r s t administered mall questionnaires in his studies then followed up w ith telephone Interviews on a selected subset o f subjects. While the questionnaire was designed w ith the Idea o f masking the in te n t o f the In v e s tig a tio n , the telephone In q u iry s p e c ific a lly focused on the exposures of p e rtine nt In te re s t, which Included 2,3,5-T and 2,4-D. Interviewers were given extensive Information on the purpose o f the study and could e a s ily have known which subjects were s o ft tis s u e sar coma cases. Interview ers probed respondents c a re fu lly only about occupations 1n which exposures to TCDD were l i k e l y . 38 0013456 4779 PON215852V -19- Recall bias is also involved in th a t workers are u n lik e ly to remember w ith accuracy the chemicals they encountered some years in the past, and i t is also extremely d i f f i c u l t to estimate the extent or duration of the exposures. I t should be noted th a t out o f 15 chemicals evalu ated by Dr. H ardell, only one, sodium c h lo ra te , did not show an elevated r is k . Nor is i t known what elevations would have been shown fo r these other agents i f they had been probed fo r in the targeted approach as fo r 2,4,5-T and 2,4-D. A dditional problems are posed by the rareness and d iv e rs ity o f these tumors. Few pathologists have been able to achieve a high degree of consistency in th e ir diagnoses.*** F urther, the id e n tific a tio n and c la s s ific a tio n of so ft tissue sarcomas have not been consistent over tim e. The sign ifica nce o f th is la s t point is Illu s tra te d by fin d in g s announced by Dr. M arilyn Fingerhut, an epidem iologist fo r the National In s titu te fo r Occupational Safety and Health (NIOSH), at an October 1983 dio xin conference at R ockefeller U n iv e rs ity . P rio r to the conference, seven cases of s o ft tissue sarcoma had been reported in the lite r a tu r e among United States chemical plant employees w ith presumptive exposures to herbicides. These reports seemed to support the fin d in g s o f D r. H ardell. At the conference, however, a d iffe re n t p ictu re emerged. NIOSH announced, a fte r detailed evaluation of the employees' work h is to rie s and medical 0013457 -20- records and microscopic examination o f the tumors, th a t two o f the seven had been misdiagnosed: the two deaths were due to carcinomas, not s o ft tis s u e sarcomas. In a d d itio n , o f the remaining fiv e , three were judged by NIOSH not to have had documented TCDD exposure. F urther, n e ith e r o f the remaining two cases had been exposed to 2,4 ,5-T but rather to the feedstock tric h lo ro p h e n o l. Furthermore, both of these cases had been exposed during upset operating conditions in v o lv in g r e la tiv e ly high TCDD exposures s u ffic ie n t to cause chloracne. Both o f remaining cases were malignant fib ro u s histiocytom as. At present, there is a growing body o f lite r a tu r e which does not support D r. H ardell's hypothesis. New Zealand: No association between the use o f these phenoxy m herbicides and s o ft tis s u e sarcoma has been found 1n an ongoing case co n tro l study 1n New Zealand, where herbicide a p p lic a tio n 1s a registered profession. According to prelim inary re su lts published 1n 1982, not one of the s o ft tis s u e sarcoma p a tie n ts had ever worked as a licensed a p p lic a to r o f these herblcldes.41 Washington State: No consistent patterns o f death due to s o ft tis s u e sarcomas were found among occupations re la ted to the use o f these h e rb icid e s. The two occupations found to be a t highest ris k in th is 1982 review were marine engineers and bankers.*? 0013458 V "p-41r 17 M M ? IS 9 5 ? -21- F inland: No s o ft tissue sarcomas were found among 1,900 Finnish -> <-_> herbicide applicators in th is 1982 review. Nor was the death rate f o r these applicators d iffe re n t from any natural cause 1n comparison w ith the Finnish to ta l male p o p u la tio n .^ Midland, Michigan: A s t a t is t ic a lly s ig n ific a n t excess of connective s o ft tissu e cancer has been found by the Environmental P rotection Agency among white females 1n Midland County (where Dow has production f a c i l i t i e s ) . The excess amounted to 13 deaths over a 20-year period. In some cases onset of the disease had begun before the woman moved to Midland. In Its 1983 p re lim in a ry re p o rt, the Michigan Department of P ublic Health (MDPH) was unable to lin k the excess w ith any environmental fa c to r, In clu ding TCDD. As part o f th is study the MDPH reviewed data fo r 28 other counties in which TCDD or other dioxins were lik e ly to be produced as a chemical manufacturing contaminant. No Increase 1n connective and s o ft t i s sue cancers was found In these counties versus those th a t did not have In d u s tria l manufacturing sources suspected o f generating such d io x in s .44 Research continues on the problem by the MDPH, funded by a $250,000 grant by Dow to the state o f Michigan. Contrary to expectations based on D r. H a rd e ll's fin d in g s , even 1n Sweden t recent reports from Stockholm's Karol1nska In s titu te In d ica te th a t Swedish fanners, about 15 percent of whom use the herbicides H ardell 0013459 4782 bow 21551% -22- probed fo r , have experienced s lig h t ly fewer cases o f s o ft tissu e sarcoma than expected, in comparison w ith the ove ra ll Swedish d a ta .4 The only way th a t Dr. H a rd e ll's fin d in g s and those of the Karollnska In s titu te could both be correct would be i f those Swedish farmers not exposed to these herbicides developed s o ft tissu e sarcomas at somewhere between one quarter and one h a lf o f the re st o f the Swedish popula tion. This seems u n lik e ly . In addition to the above, i t should also be noted th a t no deaths from s o ft tissue sarcomas were noted in the U.S. A ir Force Ranch Hand m o rta lity study o f servicemen who sprayed Agent Orange in Vietnam;44? nor were any cases o f s o ft tis s u e sarcoma noted 1n the subsequent m orbidity study. F u rthe r, examinations by the U.S. Veterans' Admini s tra tio n o f 85,000 s e lf-s e le c te d Vietnam veterans have found fewer cases o f s o ft tissu e sarcomas than would be expected from the national average.4 In the la t t e r case, however, documented exposure to Agent Orange is u n ce rta in ; and the re g is try was not Intended as an epidemi o lo g ica l study. F in a lly , the hypothesis th a t an association e xists between TCDD exposure and s o ft tis s u e sarcomas 1s not supported by research on laboratory animals: animals fed TCDD during lab ora tory experiments did not develop s o ft tis s u e sarcomas. A dditional Research 1s underway in the United States and elsewhere which should provide a d d itio n a l perspective on the s o ft tis s u e sarcoma hypothesis. '013460 -23- rit MSDSIZHM Reproduct 1ve E ffe cts Laboratory te s ts have shown th a t c e rta in levels o f TCDD can cause a v a rie ty o f reproductive disorders in pregnant animals. In pregnant mice c e rta in le ve ls o f TCDD can cause b irth d e fe c ts . In pregnant rats exposure to c e rta in leve ls can be to x ic to the fetus causing conditions such as low b ir th w eight. In pregnant women, however, studies to date have found none o f these e ffe c ts from TCDD. In a d d itio n , there is no medical or s c ie n tific support fo r claims th a t to x ic e ffe c ts in a male from exposure to TCDD could Impact a developing fetus in the female. The most w idely p u b licize d claims th a t the spraying o f herbicides contaminated w ith TCDD could cause reproductive disorders stems from the 1978 Alsea I I study by the Environmental P rotection Agency (EPA) which reportedly found a lin k between herbicide spraying and spontaneous abortions in Alsea, Oregon. But th is study has been severely and almost u n iv e rs a lly c r itic iz e d in the s c ie n t if ic comnunlty. At lea st 18 sepa ra te c ritiq u e s o f the study have found th a t I t s conclusions are not sup ported by I t s da ta.4 While some o f these are b r ie f assessments, the longest 1s a 100-page report by an In te rd is c ip lin a ry task force at Oregon S tate U n iv e rs ity , which concluded th a t, " I f there 1s a re la tio n ship between herbicide use and miscarriages 1n the 'Alsea B a s in '. . . , 1t 1s not apparent and cannot be tested using the data from the Alsea I I s tu d y ."5 More re c e n tly , EPA's handling o f the In cid e n t also came under c r itic is m by the Agency's Inspector general.51 Among the lim ita tio n s , 4734 0013461 IP s C i? < *o q . -24- of the Al sea I I study was the lack o f evidence th a t the affected women ever came in contact w ith the h e rbicide . In a d d itio n , there was no dose/response e ffe c t: miscarriages did not increase s u b s ta n tia lly when herbicide a p p lica tio n was doubled. The EPA's 1979 A1sea I I study fin d in g s resulted in the suspension of the major uses of the herbicide 2,4 ,5-T . Dow no longer manufactures 2,4,5-T in the United S tates, (A p a r tia lly owned Dow subsidiary manufactures 2,4,5-T in New Zealand.) The fo llo w in g studies were conducted to determine whether TCDD causes reproductive disorders in humans. None of the studies support the conclusions o f Alsea I I . Yarraa D is t r ic t , A u s tra lia : This 1978 in v e s tig a tio n by the - A ustralian Commission o f Public Health fa ile d to fin d any lin k between the use of 2,4,5-T and 2,4-D and b ir th defects in people o r domestic anim als.5^ Arkansas, 1948 to 1974: This 1979 In v e s tig a tio n found no association between 2,4,5-T and fa c ia l c le f t defects 1n c h ild re n .5^ Long Island Railroad: This study by the National I n s t it u te fo r Occupational Health and Safety (NIOSH), also conducted 1n 1979, found ho d e fin ite excess o f b ir th defects re la te d to 2,4,5-T exposure 1n maintenance workers exposed to the h e rb ic id e , which was used fo r weed con trol along the ra ilro a d tra c k s . 54 C0134S2 4785 21585 Hungarian A g ric u ltu re and F orestry Workers: This 1980 study found th a t despite a large increase in the use o f 2,4,5-T in th a t nation between 1969 to 1975, le v e ls o f various b irth defects e ith e r remained stable or de clin ed.55 New Zealand 2,4 ,5-T A p p lic a to rs : This 1982 study conducted by researchers at the W ellington C lin ic a l School o f Medicine found no s t a t is t ic a lly s ig n ific a n t increase in re la tiv e ris k among male professional 2,4,5-T sprayers ',,i terms o f b ir t h defects or m iscarriages. Wives involved w ith spraying a c tiv itie s or who washed contaminated clothes also had no detectable reproductive e ffe c ts . 56 Midland, Michigan: This 1983 In v e s tig a tio n by the Michigan Department o f Public Health (MDPH), updating a previous re p o rt on an Increase of b ir th defects 1n Midland County from 1971 to 1974, found th a t the Increases "may not have been unusual" and specu lated th a t the flu c tu a tio n s may have been due to changes 1n c la s s ific a tio n and re p o rtin g o f b ir th d e fe c ts .5? Further lig h t 1s shed on the Issue o f reproductive disorders and the p o te n tia l fo r to x ic e ffe c ts o f TCDD to be tra nsm itte d from the male to the fetus by a study conducted by Dow Chemical on the wives o f employees * Involved 1n herbicide production. In th is 1982 study, the te s t group consisted o f wives o f employees w ith p o te n tia l TCDD exposure, w hile the p- 4786 0013463 I -26- control group consisted o f wives o f employees w ithout such p o te n tia l exposure. One phase of analysis de alt w ith p o te n tia l e ffe c ts on f e r t i l i t y . The average number of pregnancies among wives o f 370 employees w ith poten t i a l exposure to TCDD were compared to those among wives o f 345 employees w ithout such presumed p o te n tia l exposure. Results showed no decrease in pregnancies among the te s t group: in fa c t, the te s t group had more pregnancies than the c o n tro l.58 Another phase of analysis examined m iscarriages, s t i l l b i r t h s , b ir th defects and in fa n t deaths in terms o f p o te n tia l TCDD exposure. (In th is phase, i t was necessary to redefine the b irth s in the te s t and Control groups, because some pregnancies in the te s t group took place before the fa th e r's presumed d io x in exposure. This required the tra n s fe r o f 766 b irth s from the te s t group to the c o n tro l.) Analysis found no s t a t is t ic a lly s ig n ific a n t associations between TCDD exposure and pregnancy outcomes.59 Synergism and TCDD Unsubstantiated claims have been made th a t TCDD exposure could account fo r any number o f 111 e ffe c ts 1n humans, because o f I t s a b ilit y to enhance the to x ic e ffe c ts of other compounds. A Dow Chemical m o rta lity 4787 0013464 -27- N M 2158 report on 8,181 employees working in a large production complex in v o lv ing as many as 500 d is tin c t chemical processes found no observable adverse e ffe c ts . This 24-year follow -up from 1954 to 1978 found overall m o rta lity to be 19 percent less than th a t expected fo r the correspond ing U.S. population. While the heal thy-worker fa c to r could have played a part here, i f low -le vel TCDD exposure could cause a v a rie ty of increased illn e s s e s , an increase 1n m o rta lity should have been found, not a decrease. AGENT ORANGE AND THE VETERAN On the basis of the considerable knowledge o f the human health Impact o f TCDO gained from instances o f h ig h -le ve l In d u s tria l exposure and long-term occupational exposure, 1t might seem s u rp ris in g th a t eig ht years a fte r the herbicide program 1n Southeast Asia ceased health claims were f ile d by veterans a lle g in g a lin k between th e ir current health status and what they believe to have been exposure to Agent Orange. An excess In to ta l cancers has not been seen 1n workers exposed to high le ve ls o f TCDD a t N 1tro, West V irg in ia ; Amsterdam, the Nether lands; or Times Beach and Im p e ria l, M issouri. Nor have studies found reproductive e ffe c ts from low -level TCDD exposures as a re s u lt o f he rbi cide spraying 1n A u s tra lia ; Hungary; New Zealand; o r Times Beach and Im p e ria l, M issouri; nor from male ra ilro a d worker exposures on Long Is la n d , or from the exposures o f male herbicide production workers 1n Midland, Michigan 0013465 4788 -28- F urther, male mice fed simulated Agent Orange containing two parts per m illio n TCDD (the average TCDD concentration in the Agent Orange used in Vietnam) did not show any reproductive disorders when mated w ith female mice a fte r e ig h t weeks o f e x p o s u r e . ( S e e Appendix C.) In a d d itio n , in contrast to other documented cases o f TCDD poisoning, in which chloracne serves as an in d ic a to r of overexposure, no cases of chloracne have been documented by government physicians in any o f the Vietnam veterans examined.4** On the basis of the a va ila b le in fo rm a tio n , Agent Orange does not seem e ith e r a very lik e ly or p la u s ib le explanation fo r the health problems experienced by some veterans and th e ir fa m ilie s . Nor is there any s c i e n t if ic documentation to date th a t the health o f Vietnam veterans as a 'group is any d iffe r e n t from th a t of the general popula tion. Nor u n til now has any concerted s c ie n t if ic e ff o r t been made to determine 1f the veterans o f the Vietnam c o n flic t o r any other war have suffered any long-term health problems from any wartime experiences. The p la in t if f s in the present lit ig a t io n represent less than one-tenth of one percent o f the 2.8 m illio n veterans who served in Vietnam. Many of these p la in t if f s are very i l l , from undetermined causes, and need help. While th e ir problems are real and le g itim a te , th e ir lin k to Agent Orange has y e t to be established. 0013436 4789 - 29- * tSSSSI2RS4 There are about 95 ongoing state and federal studies re la tin g to TCDO and it s p o te n tia l health impact; however, few of these studies have relevance to the Vietnam veteran exposed to Agent Orange. E s s e n tia lly , only one o f these studies, which is being conducted by the Centers fo r Disease C ontrol, examines in d e ta il the p o te n tia l fo r adverse health e ffe c ts in Vietnam veterans from a v a rie ty of p o te n tia l causes other than Agent Orange. The fo llo w in g studies have already been conducted to examine the p o te n tia l fo r Agent Orange to cause harm, among Vietnam veterans a lle g e d ly exposed. Ranch Hand Studies In a 1983 study of more than 1,200 servicemen who applied Agent Orange in Vietnam and who were deemed by the U.S. m ilita r y to have had the highest'exposures to TCDD, there was no In d ic a tio n th a t h e rbicide exposure had any e ffe c t to date on th e ir m o r ta lity .4? These servicemen are estimated by the m ilita r y to have had exposure* to Agent Orange la s tin g from ten to 12 hours a day, fiv e to s ix days a week, fo r periods o f at le a s t a year.2 ach 0f these servicemen was matched in th is study w ith fiv e other comparisons based on s im ila r jo b , race, age and month o f b ir t h , wherever p o ssib le . In a d d itio n , the Ranch Handers were compared w ith the 1978 Department o f Defense N o n d ls a b lllty Retired L ife Table and the m o rta lity experience o f the West Point class o f 1956. 0013467 4790 } -30- Results showed no adverse m o rta lity trends among the Ranch Hand veterans. The Ranch Hands were found to have 40 percent fewer deaths than expected, compared to the s ta tis tic s fo r U.S. w hite males. While chloracne has been the most consistent fin d in g among workers exposed to high levels of TCDD, a m o rbidity study re cently completed on the Ranch Hands has found no chloracne in the group. While s o ft tissu e sarcomas and porphyria cutanea tarda have been hypothesized as possible adverse outcomes of TCDD exposure, the recent study noted n e ith e r o f these disorders among the Ranch Hands nor was there any excess in to ta l cancers. Some differences did e x is t between the Ranch Hands and the c o n tro l, however. Among these fin d in g s were apparent increases in s e lf-re p o rte d neonatal deaths and minor b ir th defects and an excess o f nonmelonoma skin cancers. While these fin d in g s m e rit fu rth e r In v e s tig a tio n o f the p o te n tia l health impacts o f the Vietnam experience, 1t should be noted th a t no exposure/response re la tio n s h ip w ith Agent Orange could be e sta b lish e d . That 1s, those Ranch Hands w ith the highest presumed exposures to Agent Orange did not have a greater frequency o f disease, neonatal deaths or b irth defects when compared to less-exposed Ranch Hands. * F u rth e r, 1t should be noted th a t because the neonatal deaths and b irth defects were s e lf-re p o rte d , medical confirm ation is needed before they 4791 ^^13488 S8SURM -31- can be adequately in te rp re te d . In a d d itio n , the skin cancers were o f a type th a t is h ig hly curable and which appears to be re la ted to s u n lig h t. That is , geographical areas w ith greater su n lig h t exposure have more cases o f these skin cancers. The A ir Force study notes th a t these apparent excesses of skin cancers among Ranch Hands have not been adjusted fo r su n lig h t in terms o f geographical lo c a tio n of residence.^?. A u stra lia n B irth Defect Study According to th is 1983 study, commissioned by the A u s tra lia n M in is try fo r Veterans' A ffa ir s , no evidence was found th a t army service 1n V ie t nam has Increased the ris k o f b ir th de fe cts. The study was conducted by searching the records o f 34 A ustra lian h o sp ita ls and cytogenetics la b o ra to rie s fo r b ir th d e fective ch ild re n born to any o f th a t cou ntry's veterans who served in Vietnam between 1962 and 1972. The study "gives persuasive evidence th a t Vietnam service has not been associated w ith any Important Increase 1n the ris k o f b ir th defects 1n c h ild re n o f veterans." 63 A s im ila r study o f (J.S. Vietnam veterans liv in g near A tla n ta , Georgia, is scheduled to be released 1n spring o f 1984. 0013489 4792 A u stra lia n Senate Standing C o w itte e Report "No convincing evidence" was found in th is 240-page report o f the A ustra lian Standing Committee on Science and the Environment in it s in v e s tig a tio n o f the p o te n tia l fo r Agent Orange to cause harm th a t the rates of b ir th defects or m o rta lity were excessive among Vietnam veterans. F urther, the Committee concluded th a t Agent Orange is an improbable cause of b irth defects in c h ild re n of Vietnam veterans. "The Committee b e lie ve s ," the report s ta te s , "th a t there is no b io lo g i c a lly p la u sib le mechanism whereby the fa th e r's exposure in Vietnam can lead, years la te r , to exposure of the fetus in the u te ru s, as would be required to produce terato gen ic b ir th abnorm alities." 4 The report also noted the " s tr ik in g s im ila r itie s o f the veterans' disorders in comparison w ith those found among veterans o f World War I , World War I I , the Korean. War, and the A ra b /Is ra e li Wars."4 No conclusion on ca rcin o g e n icity was reached by the re p o rt; the Committee believed there was in s u ffic ie n t evidence to make an assessment e ith e r way.4 U.S. Agent Orange Registry Findings No "unusual long-term m o rbidity or m o rta lity associated w ith Vietnam service or Agent Orange exposure" was found in th is recent Veterans' 4793 0013470 MSSSUMM -33- A dm inistration evaluation o f 85,000 Vietnam servicemen who presented themselves to the agency fo r examination. While th is assessment cannot be considered a fu ll-fle d g e d epidem iological study, i t should be noted th a t "o f the several thousand veterans complaining o f dermatologic problems, only one may possibly turn out to be chloracne."4 While the report notes " a wide v a rie ty o f health problems," they were "o f the s o rt one sees in a population o f males growing o ld e r. National Health S ta tis tic s and the Veteran The studies ju s t cited attempt to determine i f the health e ffe c ts of Vietnam veterans can be related to th e ir p o te n tia l exposure to Agent Orange. Results of these studies should be encouraging to the veteran, and as noted previou sly, fu rth e r study continues. S ta tis tic s in d ic a te , however, th a t serious medical conditions do occur in the normal course o f events. Given a large enough popula tion, these conditions occur with s ta tis tic a l re g u la rity . P resently there is no c e rta in ty th a t the 2.8 m illio n Vietnam veterans as a group are, or are not s ic k , or dying from unusual causes or at unusual ra te s . There is some c e rta in ty , however, what the v ita l s ta t is t ic s o f a group the same s iz e , age, and sex as the veterans would be: th a t is , national health s t a t is t ic s suggest how many people 1n a s im ila r population would die w ith in a given tim e frame, and from what causes 0013471 IhSlSlZ IAQQ -34- On the basis of previous research, over the ten year period 1970 to 1979, 48,592 veterans would have been expected to d ie . Accidents, homicides and suicides would be expected to account fo r the la rg e s t p o rtio n of these deaths: 30,972. An a d d itio n a l 17,620 deaths would occur from various physical disorders: Heart disease and stroke ..................... 4973 Cancer . . . . . . . . . . . . .................. 3819 Disease of the digestive systea .................. 2109 Diseases of the respiratory systea . . . . . . . 1193 Mental d i s o r d e r s ............................ 1010 Diseases of the nervous systea . . . . . . . . . 817 Diseases of the endocrine systea . . . . . . . . 633 - Other physical disorders ................. . . . 3066 I t should be noted th a t these are s ta t is t ic s based on the m o rta lity o f U.S. w hite males/ 6 These are deaths th a t could be expected to occur In the "normal" course of events. In e ffe c t, what the study suggests 1s th a t many o f the claims being made by the p la in t if f s 1n the present lit ig a t io n are no d iffe re n t from the types o f ailm ents th a t would be expected in a s im ila r group from the general p o pula tion. 4795 0013472 ^( itsssm toa -35- Recent Developments In Ju ly 1983, TCDD was described as a probable human carcinogen by a panel called by the Environmental P rotection Agency (EPA) to review the lite r a tu r e on the compound, A primary reason fo r th is determ ination was the presence of seven cases of s o ft tis s u e sarcoma in a re g is try maintained by the National In s t it u te fo r Occupational Safety and Health (NIOSH) o f about 6,000 workers p o te n tia lly exposed to TCOD. However, in October of the same yea r, NIOSH announced th a t three o f th is seven did not have s ig n ific a n t d io xin exposure, and two o f the remaining four had been misdiagnosed: they had died o f carcinomas, not s o ft tis s u e sarcomas. N either EPA's nor NIOSH's fin d in g s 1n th is regard have been published yet in the s c ie n tific lite r a tu r e . In September 1983, the Supreme Court o f Nova Scotia ruled 1n fa v o r of the continued use o f 2 ,4 ,5-T 1n th a t province, denying p l a in t i f f s an in ju n c tio n to prevent the spraying. According to J u s tic e D. M erlin Nunn, "This court 1s o f the opinion th a t these spraying operations can be ca rrie d out 1n safety and w ithout ris k to the c itiz e n s o f th is pro v in c e ." In his decision J u s tic e Nunn c r itic iz e d witnesses fo r the p la in t if f s fo r partisanship and refusing to accept c r itic is m o f studies supporting th e ir points o f view. By c o n tra s t. J u s tic e Nunn stated th a t he did not detect partisanship on the pa rt o f the defendant, Nova Scotia Forest In d u s trie s , and accepted the evidence o f the defendant's witnesses as representing the generally accepted view o f responsible s c ie n tis ts . 4796 0013473 3 ` tiMMMUID? . -36- In December 1983, a group o f s c ie n tis ts attending an in te rn a tio n a l conference on TCDD a t Michigan State U n ive rsity concluded th a t humans appear to be less se n s itiv e to TCDD than some lab ora tory animals, and th a t in s u ffic ie n t evidence e xis ts to in d ica te TCDD is a human carcinogen. While the re p o rt noted the p o te n tia l fo r TCDD to cause reproductive e ffe c ts in pregnant women exposed to high le v e ls , such as the most contaminated areas o f Seveso, I t a ly , i t is also noted th a t there is no evidence th a t TCDD can cause b irth defects through paternal exposure. According to press accounts, researchers attending th is conference stated th a t TCDD does not warrant "an exce p tio n a lly high p u b lic p o lic y p r io r it y which d iv e rts resources and p u blic a tte n tio n from other more widespread and dangerous compounds." However, the announcement noted th a t "hot spots" where major concentrations o f TCDD e x is t should continue to be studied. Conclusion Considerable research is being dire cted towards determining whether TCDD at trace le v e ls poses a ris k to human h e a lth . A considerable amount o f research has already been conducted. This research suggests th a t: 1 TCDD causes chloracne, the most consistent in d ic a to r o f a to x ic exposure, at re la tiv e ly high le v e ls ; 4737 0013474 37- - 2. A t higher le ve ls o f exposure, TCOD causes tra n s ie n t liv e r and nerve dysfunction; no cn 3 . TCDD does not adversely a ffe c t human reproduction from to x ic exposures sustained by the fa th e r; 4 . No deaths from TCDD exposure have been documented, nor has exposure to the compound been shown to Increase the ris k of dying a t some la te r period, whether from a l l diseases in aggregate o r to ta l cancers; and 5. More research Is needed to c la r if y the re la tio n s h ip , i f any, between TCDD exposure and s o ft tissu e sarcoma. Based on the conclusions o f th is research, Agent Orange exposure as a re s u lt o f m ilita r y spraying to pro te ct American servicemen In Vietnam does not seem a lik e ly cause o f the health e ffe c ts presently suffered by some veterans and th e ir fa m ilie s . Although p re lim in a ry fin d in g s to date should be encouraging to veterans, s c ie n t if ic research Is in s u f f i c ie n t to determine d e fin itiv e ly whether the health o f Vietnam veterans as a whole is any d iffe r e n t from th a t o f the general p o pula tion. Fur th e r research is needed, and is now underway, on the p o te n tia l health impact o f the e n tire Vietnam experience on our veterans, who endured tough, demanding duty in the service o f th e ir country. 4798 0013475 SHBSIHq -38- APPENDIX A A BRIEF HISTORY OF THE AGENT ORANGE CONTROVERSY H erbicide Use In Vietnam Agent Orange, the United States m ilita r y code name fo r a 50/50 m ixture of herbicides 2,4,5-T and 2,4-D was used during the Vietnam War from 1965-1970 to d e fo lia te ju n g le vegetation and to p ro te ct U.S. servicemen from enemy ambush. During th a t time i t is estimated th a t 10 percent of Vietnam was d e fo li ated w ith herbicides and 60 percent o f th a t area was sprayed w ith Agent Orange. (In terms o f perspective, 1t might be noted th a t in 1982 the gypsy moth d e fo lia te d e ig h t m illio n acres 1n the Northeastern states according to the U.S. Forest Service -- tw ice as many acres as were d e fo lia te d during the e n tire Vietnam War.) Agent Orange was usu ally sprayed from C-123 (f1xed-w1ng) m ilit a r y a ir c r a f t . A small amount o f Agent Orange was applied from ground sources such as boats, tru cks and backpack sprayers. The to ta l amount of Agent Orange used during the h e rb icid e program 1s estimated a t between 10 and 12 m illio n g a llo n s .6^ Studies In d ica te th a t only about s ix percent o f the Agent Orange sprayed over Vietnam ever penetrated th e dense f o l i a r canopy to reach the ju n g le f lo o r ; and o f the 94 percent caught 1n the dense fo lia g e , d ir e c t exposure 0013476, rtS 8su jtM -39- to su n lig h t would have degraded most o f the TCDO w ith in 24 hours.^ The remaining TCDD which might have reached the jungle flo o r would have been t ig h t ly bound to the s o il and the refore been less ava ila ble fo r human contact. (Under these co n d itio n s, TCDO leve ls in Vietnamese s o il from Dow-supplied Agent Orange would have been more than 100 times less than the level o f concern recently set at Times Beach by the Centers fo r Disease Control fo r re s id e n tia l areas. See Appendix D.) Development of Agent Orange During World War I I , the United States m ilita r y began studying the use o f 2,4,5-T and 2,4-D, during f ie ld t r i a ls to determine th e ir c a p a b ility in d e fo lia tin g large areas of vegetation in tro p ic a l battlegrounds. M ilita r y experiments w ith these herbicides continued through the 1940s and 1950s. In 1961, a d e fo lia n t s im ila r to Agent Orange was selected by the U.S. Secretary o f Defense to be tested as the herbicide o f choice fo r use 1n Vietnam. In the e a rly 1960s the O ffic e o f the Surgeon General, Department o f the Army, conducted research on the tra ce contaminant TCDD. The published re s u lts acknowledged the work o f German s c ie n tis ts D rs. Klmmig and Schulz who observed cases o f chloracne among workers engaged in the pro duction o f 2 ,4 ,5 -J from tric h lo ro p h e n o l in the la te 1950s. Klmmig and Schulz had reported TCDD to be a contaminant o f th is manufacturing process.70 4800 0013477 mi -40- In 1962, the U.S. Government published research sponsored by the Research and Development D iv is io n of the O ffic e o f the Surgeon General, Department of the Army and by the U.S. Public Health Service which reported on the process to determine the existence o f TCDD in c e rta in chemical products. This method, known as the ra b b it ear te s t, was designed by Dow to measure skin response (chloracne) from TCDD exposure, as well as from other chemicals. In 1965, the U.S. m ilita r y decided upon a 50/50 m ixture o f 2,4,5-T and 2,4-D which la te r came to be known as Agent Orange. The m ilita r y subsequently determined the rates and frequencies of Agent Orange a p p lica tio n throughout the Vietnam herbicide program as w ell as the areas to be sprayed. As used in Vietnam Agent Orange has not been prescribed fo r any domestic weed control a p p lic a tio n s . Dow supplied about 32 percent o f the Agent Orange to the U.S. Government fo r the war e f f o r t . As a q u a lity con trol measure Dow analyzed the 2,4,5-T 1n a ll shipments of the Agent Orange i t supplied the government to Insure the absence -- no detectable le v e l -- o f TCDD, which given the technology o f the day meant less than 0.5 p a rt per m illio n in the Agent Orange. Agent Orange Lawsuits In e a rly 1979, the f i r s t of m u ltip le law suits was f ile d against seven manufacturers: Dow, the Monsanto Company, Diamond Shamrock 'J)-46<r7 4801 0D13478 It S8 S I I M M -41- Corporation, Uniroyal In c ., T.H. A g ric u ltu re and N u tritio n Company, Thompson Chemical, and Hercules In c . a lle g in g a connection between the spraying of Agent Orange in Vietnam and various maladies reported by some Vietnam veterans and th e ir fa m ilie s . These claims have been consolidated in to a class action s u it in which thousands o f Vietnam veterans and th e ir fam ily members w i l l be repre sented by selected cases. These model cases w ill be tr ie d in d iv id u a lly to determine whether Agent Orange has caused health problems to these veterans and th e ir fa m ilie s . The t r i a l is scheduled to begin in early May 1984 in t h e courtroom o f C hief Judge Jack B. Weinstein o f the Federal D is t r ic t Court fo r the Eastern D is t r ic t o f New York. In the meantime, more than 95 studies are underway to determine the v a lid ity o f health claims in v o lv in g d io xin and Agent Orange. The re s u lts o f these and other ongoing studies should help to address public anxiety over the alleged lin k between Agent Orange and human health problems. 4802 0013479 -42- APPENDIX B DOWZ \ 5 *5 49 CONSPIRACY OF SILENCE CONTENTION IN THE AGENT ORANGE CONTROVERSY Much of the present p l a in t i f f s ' case has centered around the so-called " conspiracy o f silence " claim . In b r ie f, th is theory claims th a t Dow knew th a t Agent Orange was harmful to users and met w ith other chemical producers d e lib e ra te ly to conceal th is inform ation from the government. The theory does not hold up under s c ru tin y . An examination o f the chronology of events surrounding the se le ctio n o f Agent Orange by the m ilita r y fo r use in Vietnam shows 1) th a t the United States government had extensive knowledge o f both the components of Agent Orange as well as the p o te n tia l occupational health e ffe c ts o f the contaminant TCDD; 2) th is knowledge on the pa rt o f the government was concurrent w ith th a t o f Dow and the other co-producers; and 3) th a t Dow's concerns about TCDD centered around the p o te n tia l o f r e la tiv e ly high le ve ls o f the compound to cause harm In the workplace not on the end uses o f the fin ish e d product which contained v a s tly lower le v e ls o f the contaminant. Chronology 1949 The U.S. Public Health Service (USPH) In ve stig a te s the chloracne Incident a t a trlchlorophenol production plant at N 1tro, West V irg in ia . 4 o no 48 03 00134BO -43- W H I I 58531 1957 Having determined TCDD to be the cause of chloracne in it s trich io rop he nol workers, the German firm Boehringer Sohn sends a le t t e r to Dow Chemical regarding the a v a ila b ility of technology to address the problem. Dow has no chloracne problem at the time of re ce ip t o f the inform ation, and the le tte r is file d fo r future reference. 1959-60 United States m ilita r y personnel at Edgewood A rsenal, F o rt D e tric k , and the Anmy Corps Research and Development Laboratory become aware of the chloracne p o te n tia l in trichiorophenol manufacture, through the research o f two German s c ie n tis ts , Kimmig and Schulz. TCDD is considered b r ie fly by the m ilita r y as a weapon o f war. An a r tic le published by the USPHS ind ica tes knowledge th a t chloracne was associated w ith 2,4,5-T production. 1962 M ilita r y personnel at F o rt D e tric k recommend a d e fo lia n t form ula tion, la te r to become known as Agent Orange, fo r use in Vietnam. Research supported by the Research and Development D iv is io n o f th e O ffic e o f the Army Surgeon General Indicates both knowledge o f TCDD's acnegenic p o te n tia l and th a t the contaminant could he produced a t high temperatures from t r i chi orophenol. 0013481 4804 J)- -44- |>OW 2153531 1*363 The USPHS completes in v e s tig a tio n of a chloracne incide nt at a New Jersey 2,4,5-T production p la n t. 1964 A process change in Dow Chemical trichlorophenol production at Midland, Michigan, exposes 61 workers to high levels of TCDD. Of those exposed, 49 workers have some evidence of chloracne. The trich lo ro p h e n o l plant is shut down fo r a period to resolve the problem. TCDD is f i r s t suspected, then confirmed, as the cause of the problem. 1965 Dow establishes an in te rn a l standard fo r detectable TCDD at an a n a ly tic a l lim it of one p a rt per m illio n . Dow c a lls a meeting w ith its competitors to discuss the presence of the acnegen in tric h lo ro p h e n o l. Dow also informs the USPHS, the Michigan State Department of Public Health, the University of Michigan Institute for Industrial Health and oth er Michigan u n iv e rs itie s , the United States Army Biological Laboratories at Fort Detrick, the Ontario Department of Labor, and other government agencies o f the Midland trichlorophenol chloracne In c id e n t. Also n o tifie d we/*e a number of physicians and health o f f ic ia ls from p riv a te in d u s try , and from fo re ig n countries in clu d in g the Netherlands, England, South America and the Soviet Union. I 4805 00B : 2 -45- 01215855* M ilita r y personnel- at Fort Detrick request t o x ic it y data on TCDD from Edgewood Arsenal. Late in the year, Dow supplies the m ilita r y w ith it s f i r s t shipment of Agent Orange. 1966 A new Dow trichlorophenol plant comes on lin e at Midland, Michigan, b u ilt w ith technology licensed from Boehringer Sohn in Europe. The Army Surgeon General's Office requests t o x ic it y informa tio n on 2,4,5-T from the National Academy of Sciences (NAS); the NAS response references porphyria and chloracne as e ffe c ts o f exposure. Personnel at Eglln Air Force Base lea rn o f the p o te n tia l f o r TCDD to cause occupational harm as the m ilita r y considers con s tru c tio n o f I ts own 2,4 ,5-T p la n t, due to short supply o f Agent Orange. 1967 With regard to the government's plan to b u ild I ts own Agent Orange production f a c i l i t i e s , Dow Chemical Informs Edgewood Arsenal an$l the Department of Defense o f the "serious poten t ia l health hazard" (chloracne) to 2,4,5-T In d u s tria l plant workers. 4806 0013483 -46- The Rand Corporation reports th a t Infants could p o te n tia lly receive le th a l doses from the wartime crop-destruction pro gram. The report is passed by Secretary o f Defense Robert McNamara to the J o in t Chiefs o f S ta ff, who recommend th a t the herbicide program continue. O f, o t .1 1968 Thompson Chemical Company, in the course of meetings re la tin g to the m ilit a r y ^ plan to b u ild it s own Agent Orange f a c i l i t ie s , informs m ilita r y personnel o f the a v a ila b ility of tech nology to reduce TCDD in Agent Orange. Also discussed are the hazards o f TCDD. 1969 M ilita r y personnel at Fort D e tric k receive data, p r io r to it s o f f ic ia l release, on the te ra to g e n ic ity o f 2,4 ,5-T in animals based on animal studies conducted by B1onet1cs Laborato ries. A fte r the Inform ation has been made a v a ila b le to the p u b lic , the Department o f Defense orders the r e s tr ic tio n of Agent spraying to areas o f Vietnam remote from population. 1970 Dow advises Edgewood Arsenal personnel th a t the TCDD 1n the 2,4,5-T used 1n the B1onet1cs study probably accounted fo r the terato gen ic fin d in g s . Spraying o f Agent Orange in Vietnam is suspended by the government. 0 01 3 4 1971 Edgewood Arsenal personnel In d ica te th a t stockp iled Agent Orange containing less than one p a rt per m illio n o f TCDD 1s v 4807 tS S 8 S IZ N M -47- s t i l l safe fo r use by the m ilita r y . Stockpiles are u ltim a te ly incinerated at sea. Note: p la in t if f s have argued th a t the Department of Defense should have been n o tifie d d ir e c tly by Dow. Dow has no proof th a t such n o tific a tio n did or did not take place p rio r to 1967. However, Dow's concerns centered around occupational exposure to TCDD not exposure to Agent Orange or any other finished product. I t should also be noted th a t Dow was not a su p p lie r o f Agent Orange to the government at the time o f the Midland chloracne in c id e n t, and measures to c o rre c t the problem had been taken before Dow's f i r s t shipment to the government. In a d d itio n , the record shows th a t Dow n o tifie d government agencies of it s occupa tio n a l problem w ith TCDD. Such a wide sharing o f knowledge can hardly be equated w ith the so-called " conspiracy o f s ile n c e ." F in a lly , the chronology ju s t given reveals extensive knowledge on the part o f the government, and by m ilita r y personnel w ith in the government, regardless of when n o tific a tio n to the Department o f Defense by Dow did or did not take place. * 0013435 4808 -48- S S s a s r z 1* o < r APPENDIX C ANIMAL STUDIES The Toxicity of TCDD In Animal Species There are wide v a ria tio n s in how d iffe r e n t animal species are affected by TCDD. For example, the sing le oral dose LD50 (the amount th a t i t would take to k i l l 50 percent of the te s t animals) in guinea pigs is 0. 6- 2.0 ug/kg* (micrograms per kilogram o r every 2.2 pounds o f body weight [a microgram is one m illio n th of a gram; a kilogram is 1,000 grams, o r 2.2 pounds]).71*72 j n hamsters, however, TCDD is much less to x ic w ith an oral LD50 o f 1157-5051 ug/kg.^3,74 Therefore, the guinea pig is approximately 5,000 times more s e n s itiv e than the hamster, although both species are commonly used fo r lab ora tory experiments. The d ire c t relevance o f any s in g le animal experiment to people must be evaluated w ith respect to a ll other availa ble data. In acute and subchronic s tu d ie s , l iv e r t o x ic it y 1s a prominent component o f TCDD t o x ic it y 1n ra ts , mice and ra b b its , but not 1n monkeys where e ffe c ts on the bone marrow and e p ith e lia l tissu e are more promi nent. * See conversion c h a rt, Appendix E. 0013436 4809 y - w S M B# 11M -49- Thymic atrophy in ea rly to x ic ity studies suggested th a t TCDD might decrease the immune response. In lab ora tory animals, s c ie n tis ts can induce immunotoxic e ffe cts w ith high doses o f TCDD. However, as the Seveso in cid e n t shows, these e ffe c ts have not been demonstrated in exposed humans.7 *14 Teratogenicity/A nim al Species Of the various to x ic responses of animals to TCDD, the p o te n tia l fo r te ra to g e n ic ity ( b ir th defects) has perhaps received the most a tte n tio n . Teratogenic e ffe c ts re s u ltin g from TCDD exposure have been re a lize d only in mice which show an Increased frequency o f c le f t palate and a kidney abnorm ality. I t is w ell known th a t many factors can cause b ir th defects in mice including the stress o f being transported by a ir during pregnancy or being deprived o f d rin k in g water ove rnig ht. In ra ts TCDD does not cause a tru e teratogenic e ffe c t, but s u ffic ie n tly high doses can cause embryo- and fe t o to x lc ity . In both ra ts and mice, dose le v e ls o f TCDD have been id e n tifie d at which these e ffe c ts do not occur. Many o f the studies with TCDD 1n monkeys have been conducted a t the U n ive rsity o f Wisconsin. In experiments where the high dose le ve l of about 0.011 ug/kg/day* TCDD 1n the d ie t was given to the monkeys fo r up t. See conversion chart, Appendix E 0013487 f btvj z i5 8 5 5 7 -50- to 9.3 months, there were substantial to x ic e ffe c ts .75,76 ^ p relim ina ry abstract o f a follow -up study77 showed th a t monkeys given d ie ts con ta in in g approximately 0.0017 ug/kg/day* o f TCDD had only s lig h t t o x ic it y . There are studies c u rre n tly in progress at the U n iv e rs ity of Wisconsin at lower doses. In a three-generation reproduction study7** o f ra ts exposed to in c re mental dose leve ls of TCDD in the d ie t high doses caused decreased f e r t i l i t y and decreased neonatal s u rv iv a l. The interm ediate dose leve l caused decreased f e r t i l i t y and other e ffe c ts in the f i r s t two genera tio n s but not in the t h ir d . At the low-dose leve l there was no im pa ir ment of reproductive capacity through the three consecutive generations, in d ic a tin g th a t 0.001 ug/kg/day* was a no-adverse-effect le ve l over m ultiple generations. In a d d itio n , animal studies done to date do not suggest th a t b ir th defects can be passed from the male to the female as a re s u lt o f a to x ic TCDD exposure in the male. In a study conducted by the National Toxi cology Program, laboratory mice were fed simulated Agent Orange contain ing two parts per m illio n o f TCDD (the average concentration o f TCDD In the Agent Orange used in Vietnam) fo r e ig h t weeks as p a rt o f t h e ir d ie t. D e fin ite signs of t o x ic it y , in terms o f reduced weight gain and liv e r t o x ic it y , wei^e noted in the exposed males. Yet, when these males were See conversion chart, Appendix E 00134S8 4811 i6S0SIZMi -51- mated to unexposed v irg in females, "no s ig n ific a n t decrease in f e r t i l i t y o r reproduction" was noted. Survival o f o ffs p rin g was "apparently unaffected by paternal exposure to the simulated mixtures o f Agent Orange."55 Hutagenicity/Animal Species A number o f mutagenic studies o f TCDD have been conducted w ith various in v it r o te s t systems using b a c te ria l yea st, p la n t and tis s u e c u ltu re te s ts . A dditional te sts have been conducted w ith in ta c t animals as w ell as human c e l l s . 79, 80, 81, 82, 83, 84, 85 While a few p o s itiv e or questionable mutagenic responses have been observed in ce rta in plant or m icrobial te s t systems, there appear to be no d e fin itiv e correlates o f m utagenicity in higher animals or people. Carcinogen!clty/Animal Species Carcinogenic studies fo llo w in g Ingestion of TCDD have been conducted in ra ts and mice.fi68788 Review o f these data ind ica tes th a t the re s u lts in ra ts and mice c o rre la te w ell w ith a carcinogenic response associated on ly w ith life tim e ingestion o f higher dose le v e ls th a t also Induce other t o x ic it y . The liv e r was the primary ta rg e t fo r cancer in both ra ts and mice. No cancer response occurred a t dose le v e ls of 0.001-0.0014 ug/kg/day* in the ra ts and 0.001-0.03 ug/kg/day* in mice. 0013489 p - H io r b S S iS I Z t*W -52- TCDD does cause cancer in animals. However, th is happens only at high dose le ve ls -- i . e . , dose le v e ls th a t are higher than those th a t e l i c i t other kinds o f t o x ic it y . Cancer induced by exposure to TCDD would be preceded by substantial signs of t o x ic it y . This t o x ic ity (which 1s re versible when exposure is discontinued) would act as a s e n tin e l, or warning, at dose leve ls below those th a t might cause cancer. t See conversion table, Appendix E. 0013490 -53- atsism iM APPENDIX D ENVIRONMENTAL HAZARD EVALUATION Dioxin In Soil P otential human exposure from TCDD environmental contamination concerns many people. The U. S. Centers fo r Disease Control (CDC) has set a level of concern fo r TCDD 1n re s id e n tia l s o il at one pa rt per b i l l i o n . This recommendation is based 1n part on the hypothetical cancer ris k mathematically derived from la b o ra to ry animal s tu d ie s . Im p lic it in th is recommendation is the recognition th a t the amount o f chemical absorbed in to the body determines whether a harmful response w ill occur. That 1s, dose makes the poison. Even fo r very to x ic chemicals such as TCDD th e re can be concentrations w ith in the body which w ill cause no harm. The amount o f TCDD absorbed from the s o il depends upon 1) duratio n of contact w ith the skin and 2) ingestion o f the s o il. The known s o il binding tendencies of TCDD reduces the amount th a t can be absorbed through the skin or in te rn a lly via in g e s tio n . Animal studies show th a t only a fra c tio n o f the TCDD Is absorbed from contaminated s o i l . In animals studies when such s o il contained about 0.5 p a rt per m illio n o f TCDD and the s o il was held in contact w ith skin fo r 24 hours (covered w ith aluminum f o i l ) , less than 0.2 percent of th e TCDD in the s o il was absorbed,89 0013491 4814 -54- In 1983, CDC announced a one part per b illio n level o f concern at Times Beach fo r TCDD in re s id e n tia l s o i l . 90 According to the CDC a prime consideration in e sta blishin g th is g u ideline was the p o s s ib ility of ch ild re n at play in re s id e n tia l areas who might eat d i r t . 9! Based on the COC's assumptions and c a lc u la tio n s and those o f Dr. Kingsley Stevens,92 i t appears th a t general environmental s o il contamination in the range o f low parts per b illio n would not pose a health hazard to the general population, in clu d in g the unborn, c h ild re n , pregnant women or the old and in firm . Dioxin In Fish Rats consuming a d ie t containing 22 parts per t r i l l i o n (p p t) TCDD (1 nanogram per TCDD to k ilo g ra m .o f body weight per day) fo r a life tim e showed no adverse health e ffe c ts . Rats consume about 10-20 percent of th e ir body weight in food each day, w hile people consume about two to three percent of th e ir body w eigh t. Thus, fo r a given TCDD concentra tio n in d ie ta ry components, ra ts w ill Ingest r e la tiv e ly more o f the compound than people. A person eating about two to three pounds o f food per day uniform ly contaminated w ith 25 parts per t r i l l i o n o f TCDD would Ingest about 0.5 nanogram per kilogram o f body weight per day* or about one-half the dose (weight o f TCDD per u n it o f body w eight) shown t See conversion chart, Appendix E. 0013492 4815 W tH 5 8 9 t to produce no adverse health e ffe c ts when fed to rats fo r a life tim e . Again, th is is true because people eat less food 1n proportion to th e ir body weight compared to laboratory ra ts . Most people in the United States (99 percent according to 1981 U.S. Food and Drug Adm inistration fig u re s ) eat less than 0.6 pounds o f fis h per week. Nine out of ten people in the United States eat less than 0.3 pounds o f fis h per week, w hile the "average" person eats s t i l l le s s . Since fis h is a re la tiv e ly small part o f the American d ie t, 99 percent of the American public would receive less than 1/70 o f the amount of TCDD shown to produce no e ffe c t in laboratory ra ts , fed TCDD d a lly fo r th e ir life tim e s , i f those people ate fis h containing 25 parts per t r i l l i o n of TCDD. Or said another way, a person could eat nearly one ton o f fis h per year containing 25 parts per t r i l l i o n o f TCDD and not exceed'the n o -e ffe ct level established by lab ora tory animal experiments. The fis h consumption guideline o f 25 parts per t r i l l i o n established by the Food and Drug A dm inistration assures an adequate margin o f safe ty to p ro te ct people from overexposure to TCDD.93 0013433 4816 ' j ) - n fid- - 56- TCDD Concentrations in Soil (a) Carter, e t ai.t (1975): Science, 138:738-740. (b) Faneiii, e t al., (1982): Drug Metabolism Review 13(3): 407-422. (c) Interim Report of the Missouri Dioxin Task Force, June 1,1983 (d) Kimbrough, eL al., (1983): Centers for Disease Control. GCn-191 (e) Estimate based on 0.5 parts per million TCDD in the defoliant and assuming 94% photodegredation in the jungle canopy, with remaining TCDD distributed throughout the top centimeter of soil (cf.. Young, e ta !., 1978 : U.S. Air Force, p. I-20, for documentation on photodegredation estimate. P ttm s tm -57- APPENDIX E CERTAIN UNITS OF HEASURE 1 m illig ra m (lmg) 1 gram x 10"3 O.OOl g. (1000 m illigram s - 1 gram) 1 microgram o r 1 ug * 1 gram x 10"6 0.000001 g. 1 nanogram = 1 gram x 10"9 * 0.000000001 g. 1 picogram * 1 gram x 10*12 - o .000000000001 g. 1 m illig ra m 1000 micrograms * 1 m illio n nanograms ( 1, 000, 000) * 1 b illio n picograms ( 1,000, 000, 000) 1 m illigram per kilogram or per l i t e r is 1 part per m illio n 1 microgram per kilogram is 1 p a rt per b illio n 1 nanogram per kilogram 1s 1 p a rt per t r i l l i o n 1 microgram per gram 1s 1 pa rt per m illio n 1 microgram per m i l i l l t e r 1s 1 pa rt per m illio n 1 nanogram per gram 1s 1 pa rt per b illio n 1 picogram per gram 1s 1 part per t r i l l i o n 1 picogram per l i t e r is 1 part per q u a d rillio n iq /k 1logram 1.0 0.1 0.01 0.001 0.0001 ppm 1.0 0.1 0.01 f 0.001 0.0001 _ppb ,, 1000.0 100.0 10.0 1.0 0.1 ppt 1, 000,000 100,000 10,000 1,000 100 4818 M < W 0013495 Agent Orange Carcinogen Chloracne Immunoresponse In itia to r Kilogram Malignant fibrous h istio cyto m a Microgram M orbidity Nanogram * -58- GLOSSARY APPENDIX F a 50/50 herbicide m ixture o f 2,4,5-T and 2,4-D produced according to U.S. government s p e c ifi cations fo r use as a d e fo lia n t in Vietnam. I t contained trace elements of TCDD as a re s u lt of the 2,4,5-T manufacturing process. any agent or substance which produces cancer, accelerates the development o f cancer or acts upon a population to change it s to ta l frequency o f cancer in terms o f numbers or d is tr ib u tio n by s ite and age. an acneform eruption caused by exposure to chlorinated hydrocarbons. Chloracne is considered to be the hallmark or sentinel symptom of human overexposure to the chemical contaminant TCDD. the capacity o f an organism to respond to any substance which is capable, under appropriate c o n d itio n s , o f Inducing a s p e c ific immune response and o f reacting w ith the products of th a t response, such as s p e c ific antibod ies. a carcinogen responsible fo r producing the f i r s t stage of a cancer. This step usually Involves Irre v e rs ib le mutational changes, which do not necessarily cause cancer. a u n it o f weight o f the m etric system, being 1,000 ( 103) grams, o r the equivalent of 2.20 pounds. A s o ft tis s u e sarcoma composed o f c e lls of fibrous connective tis s u e . a u n it of weight o f the m etric system, being one-m1l l 1onth o f a gram ( 10"6 gm .). or one one-thousandth o f a m illigra m ( 10"3 m g.). the condition o f being diseased. a u n it of weight of the m etric system, being one one-b1llio n t h ( 10' 9) gram. V-h v S 0013496 K9W21S(T599 -59- No-effect level the amount o f chemical th a t has been determined by te s tin g to cause no harm to the subject. Peripheral nerve Porphyria cutanea tarda (PCT) Promotion S oft tissu e sarcoma the part of the nervous system comprising the cra n ia l nerves, the spinal nerves and the sympathetic nervous system, excluding the brain (ce ntra l nervous system) and spinal cord. a member of a group of uncommon diseases, the porphyrias, th a t comprise disturbances in the body's form ation of hemoglobin, the red pigment in the blood. A s p e c ific pattern o f chemicals ca lle d porphyrins, excreted in the urine and s to o l, characterizes PCT and re fle c ts a d e fic i ency o f one of the liv e r 's enzymes involved in hemoglobin form ation. The disease manifests I t s e l f in the skin where small and large b lis te r s form on the exposed parts o f the body, probably as a slow response to s u n lig h t. The skin becomes very fr a g ile and e a s ily rubs o ff to produce sores th a t scab over and sometimes leave scars. the second stage o f the carcinogenic process which may be the re s u lt o f a d d itio n a l applica tion s of a carcinogen, or of oth er, noncarcinogenic m a te ria ls ; or i t may Involve a d d itio n a l unknown processes which could aid the growth o f the cancer or reduce resistance to the developing cancer. TCDD is among the group of chemicals th a t may secondarily in flu ence the form ation o f cancer by promotion In some s tra in s o f ra ts and mice. a r e la tiv e ly rare group o f about 100 tumors in the body's muscles, tendons and other con nective tis s u e s , 1n fa t tis s u e s , blood vessels and nerves. 0013497 ' 4820 TCDD Teratogenic Thymus T o x ic ity 2,4-D 2.4,5-T T ric h l orophefiol -60- acronym fo r 2 , 3, 7 , 8-te tra ch lo ro d ib e n zo -p d io x in , an unwanted trace contaminant formed during the synthesis of 2 ,4 ,5 -trich lo ro p h e n o l. embryonic maldevelopment leading to serious congenital defects. I . e . , causing b irth defects. a ductless gland-11ke s tru c tu re , situa ted ju s t behind the top o f the breastbone, th a t plays some part in b u ild in g resistance to disease. S' ? c*9 the inherent property o f being poisonous, e s p e c ia lly the degree to which any substance can cause acute or chronic In ju ry to liv in g th in g s , o r which is suspected o f being able to cause disease or in ju r y under some c o n d itio n s . A ll substances, man-made or n a tu ra l, are to x ic to some degree; the lik e lih o o d o f harm depends on t o x ic it y and the amount taken in to the body. also known as 2,4-d1chlorophenoxyacet1c a c id , th is chemical compound is a w idely used h e rbicide and a chemical re la tiv e o f 2 , 4 , 5 - t r ichlorophenoxyacetlc acid . A 50/50 m ixture of these two chemicals, known as Agent Orange, was used as a d e fo lia n t during the Vietnam c o n f lic t . 2,4-0 has been w idely used as a h e rbicide fo r broadleaf weed control 1n cereal crops, sugar cane and c itru s f r u i t s , and on t u r f , pastures and noncrop land. At present le v e ls o f a n a ly tic a l d e te c tio n , 2,3,7,8-TCOD has not been Id e n tifie d 1n any 2,4-D form ulations. 2,4,5-tr1chlorophenoxyacet1c acid 1s a sele c tiv e w e e d k ille r which has root Inducing, growth in h ib itin g and herbiclda l pro p e rtie s comparable to those o f the he rbicide 2,4-D (2,4-dichlorophenoxyacet1c a c id ). 2,4 ,5 -T 1s known to be p a rtic u la rly a c tiv e against woody shrubs and tre e s . I t has been used 1n fo r e s try , on grassland, f o r In d u s tria l and ric e and sugar cane weed c o n tro l. also known as 2,4,5-TCP. Used as a s ta r tin g m aterial in the manufacture of a series of In d u s tria l and a g ric u ltu ra l chemicals, the most notable o f which 1s the herbicide 2,4 ,5-T and i t s re la te d products includ ing s llv e x . Most ^ o f the TCDD present 1n 2,4,5-T 1s formed during the synthesis of trlch lo ro p h e n o l. , 48210013498 Q *8 0 S IZ K M -61- 2,3,7,8-tetrachlorodibenzo-p-dioxin also known as TCDD op d io x in . I t is formed as an unwanted contaminant during the synthesis o f 2 ,4 ,5 -tric h lo ro p h e n o l. Considered to be the most to x ic o f the 75 chemical compounds th a t make up the fam ily known as d io x in s . Also occurs, w ith other members o f the fa m ily , as a re s u lt o f the process of combustion. * C013439 4822 -62- s g jy & H a r ' INDEX Agent Orange R egistry: 32 Alsea I I : 24 Amsterdam: 15, 27 American Medical A ssociation: 4-7 Arkansas: 24 Army Corps RAD Laboratories: 43 A u s tra lia : 8, 24, 27, 31, 32 A ustra lian b irth defects study: 31 bankers: 20 BASF: 14 b ir th defects, te ra to g e n ic ity : 7, 8 , 10, 11, 15, 23, 24, 25, 26, 30, 31, 32, 36, 46, 49 Boehringer Sohn: 43, 45 Bolsover, the United Kingdom: 13 cancer, carcinogenesis: 5, 6 , 7, 8, 12, 13, 14, 15, 17-22, 27, 30, 31, 32, 34, 35, 36, 51, 52 Centers fo r Disease C ontrol: 1, 9, 29, 39, 53 chloracne: 2, 5, 10, 12, 13, 14, 15, 16, 28, 30, 33, 39, 42, 43, 44 C o a lite and Chemical Products, L td .: 13 Council fo r A g ric u ltu ra l Science and Technology (CAST): 8 Diamond Shamrock C orporation: 16, 40 Dow Chemical Company: 12, 13, 16, 17, 24, 25, 26, 40, 43, 44, 45, 46, 47 Edgewood Arsenal: 43, 45, 46 E glin A1r Force Base: 45 Environmental P rotection Agency: 21, 23, 24, 35 f e r t i l i t y : 26 f e t o t o x id t y : 23, 49 Finland: 21 fis h : 54, 55 Food and Drug A d m in istra tio n : F ort D e trick: 43, 44, 46 55 guinea pigs: 48 hamsters : 48 H a rd e ll, Dr. Lennart: 17-22 health s ta t is t ic s : 33 Hercules, In c .: 41 hexachlorophene: 15 Hungary: 25, 27 immunosuppression: 10, 13 Im p e ria l, M issouri: 15, 27 In itia tio n : 5, 6 In te rn a tio n a l Agency fo r Research on Cancer: 11 C013500 p -tfi* 4823 -63- S iS S S U M M Karolinska In s titu te : 21, 22 Kimmig and S chultz: 39, 43 LD50: 48 liv e r : 5, 10, 14, 15, 16, 48 Long Islan d: 24, 27 Ludwigshafen, Germany: 14 malignant fib ro u s histiocytom a: 20 marine engineers: 20 mice: 48, 49, 51 Michigan State U n iv e rs ity : 36 Michigan Department o f P ublic Health: 21,. 25,, 44 Midland, Michigan: 12, 21, 25, 27 m iscarriages, spontaneous abortions: 11, 23, 24, 26 monkeys: 48, 49, 50 Monsanto Company: 12, 40 m o rta lity , death: 2, 11, 12, 13, 14, 15, 29, 30, 34 m utagenicity: 6 , 7, 51 National Toxicology Program: 50 neonatal deaths, in fa n t deaths: 26, 30 New Jersey: 16 New Zealand: 20, 25, 27 National In s titu te fo r Occupational Safety and H ealth: 13, 19, 20 National In s titu te fo r Environmental Health Sciences: 11 N itro , West V irg in ia : 12, 27, 42 Nova S cotia, Supreme Court o f: 35 NV P h illip s : 15 observer bias: 18 Oregon S tate U n iv e rs ity : 23 peripheral nerve disorders: 10, 14 P h illip s , NV: (see NV P h illip s ) porphyria cutanea tarda: 16 promotion, cancer: 6 Queensland Cabinet Committee Report: 8 ra b b it ear te s t: 40 ra b b its : 48 Ranch Hands: 22, 29, 30, 31 ra ts : 48, 49, 50, 51, 52 re c a ll b ia s : 18, 19 R o cke fe lle r U n iv e rs ity : 19, 35 Seveso, I t a ly : 9-11, 27, 36 s o ft tis s u e sarcoma: 12, 13, 17-22 s o il: 53, 54 0013501 -64- Spolana, Czechoslovakia: 16 Sweden: 18, 21, 22 Stevens, Dr. Kingsley: 54 stomach cancer: 14 synergism: 26 T.H. A g ric u ltu re and N u tritio n Company: Thompson Chemical: 41, 46 thymic atrophy: 49 trlc h lo ro p h e n o l: 9, 12, 42, 43 Times Beach, M issouri: 9, 15, 16, 27 41 United Kingdom P esticide Advisory Committee: Uni ro y a l, In c .: 41 I) c A-ir Fnrre* 2? U*.s! Public Health Service: 40, 42, 43, 44 U niversity o f Wisconsin: 49, 50 7 Veterans A dm in istratio n: 22, 32, 33 Washington S tate: 20 W ellington School o f Medicine: 25 w hite women: 21 Yarram D is t r ic t , A u s tra lia : 24 t 0013502 4825 -65- USSSUftM REFERENCES 1. The Health E ffe cts of Agent Orange and P olychlorinated D1ox1n Contaminates, Amen can Medical A ssociation, October 1981. 2. Ib id . 3. Ib id . 4. Further Review of the Safety fo r Use in the U.K. o f the H erbicide 2,4,5-T, Advisory Committee on Pesticides of the M in is try of A g ric u ltu re , Fisheries and Foods (United Kingdom), February 7, 1983. 5. A Report on 2,4-0 and 2,4,5-T and Human Health, Interdepartmental Committee Appointed by Queensland Cabinet (A u s tra lia ), 1981. 6. The Phenoxy Herbicides: Second E d itio n , Council fo r A g ric u ltu ra l Science and Technology, Report No. 77, August 1978. 7. Homberger, E ., Reggianl, G ., Sambeth, J . , and W1pf, H .K ., (1979): The Seveso Accident: I t s Nature, Extent, and Consequences. Ann. Occup. Hyg. 22: 327-370. 8 . P occhiari, F. e t. al_., (1979): Human Health E ffe c ts From Accidental Release of Tetrachlorodibenzo-p-d1oxin (TCDD) at Seyeso, I t a ly . Ann. N.Y. Acad. Sc1., 311-320. 9. F a n e lli, F ., e t. al_., (1982): TCDO Contamination in the Seveso In c id e n t. Drug Metab. Rev. 13: 407-422. 10. Crow, K ., (1978): The Chemical Disease. New Scientist: A p ril 13, 1978. 11. Caramaschi, F. e t. a l . , (1981): Chloracne Following Environmental Contamination by~'rcU!T in Seveso, I t a ly . Int. J. Epidemiol., 10: 135-143. 12. Fara, G. M., et a l . , (1980); Chloracne A fte r Release of TCDO a t Seveso, I t a ly . TrT: Chlorinated Dioxins and Related Compounds: Impact on the Envlronaent, Oxford: Pergamon Press, pp. 545-559. 13. Reggianl, G ., (1979): Estim ation o f the TCDD Toxic P o te n tia l 1n the L ig h t o f the Seveso In c id e n t. Arch. Toxicol. (Suppl.), 291-302. , 14. Reggianl, G ., (1980): Acute Human Exposure to TCDD 1n Seveso, Italy. J. Toxicol. Environ. Health, 6: 27-43. D -w ? - 0013503 4826 2L SSI? AV0C7 -66- 15. B ls a n ti, L ., B onnetti, F ., Caramaschi, F ., Del Como, G., F a v a re tti, C., Giambelluca, S ., Marni, E ., Montesarchlo, E., P u c c in e lli, V., Remotti, G., Volpato, C., and Z a m brelli, E ., (1978): Experience of the Accident of Seveso, proceedings from the S ixth EST Conference: 1-32. 16. Abate, L ., e t. a l . , (1980): M o rta lity and B irth Defects From 1976 to 1979 in the l^ p u la tio n L ivin g in the TCDD P olluted Area of Seveso. In : Chlorinated Dioxins and Related Compounds: Impact on the Environment, Oxford: Pergamon Press, pp. 571-587. 17. Reggiani, G ., (1977): Toxic E ffe cts o f TCDD 1n Man. NATO Work shop in Ecology, G u ilfo rd , England, July-A ugust, 1977. 18. Zack, J . A ., and Suskind, R. R ., (1980): The M o rta lity Experience of Workers Exposed to Tetrachlorodibenzodioxln in a Trichlorophenol Process Accident. J . Occup. Med., 22: 11-14. 19. Moses, M., L i l l s , R., Crow, K. D ., Thornton, J . , Fischbein, A ., Anderson, H. A ., and S e lik o ff, I . J . , (1984): Health Status of Workers With Past Exposure to 2,3,7,8-Tetrachlorod1beno-p-dioxin in the Manufacture of 2,4,5-Trichlorophenoxyacetic A cid: Compari son o f Findings With and Without Chloracne. Am. J. Ind. Med., 5: 161-182. 20. Cook, R. R ., Townsend, J . C ., O tt, M. G ., and S llv e rs te in , L. G ., (1980): M o rta lity Experience of Employees Exposed to 2 ,3 ,7 ,8Tetrachlorodibenzo-p-dioxin (TCDD). J. Occup. Med., 22: 530-532. 21. May, G., (1973): Chloracne From the Accidental Production of Tetrachlorodibenzodloxin. Br. J. Ind. Med., 30: 276-283. 22. Hay, A ., (1982): In d u s tria l Accidents 1n Trichlorophenol Manufac tu re . In : The Chemical Scythe: Lessons of 2,4,5-T and Dioxin, Plenum Press, pp. 109-121. 23. O ffic e of Technology Assessment Review o f Two Papers Reporting on Long-Term E ffe cts of Exposure to D io x in . In : Congressional Record/Senate, September 29, 1983, S13220-S13221. 24. May, G., (1982): Tetrachlorodibenzodioxln: A Survey of Subjects Ten Years A fte r Exposure. Br. J. Ind. Ned., 39: 128-135. 25. Goldman, P. J . , (1973): Severe Acute Chloracne: A Mass In to x ic a tio n by 2,3,7,8-Tetrach1orod1benzod1ox1n. Hautarzt, 24: 149-152. 26. H u ff, J . E., Moore, J . A ., S aracci, R ., Tomatis, L . , (1980): Long-Term Hazards o f P olychlorinated D1benzodlox1ns and Poly ch lo rin a te d Dlbenzofurans. Environ. Health Perspect. 36: 221-240, 0013504 ns aeim sttw -67- 27. Thiess, A. M., Frentzel-Beyme, R., and L in k , R ., (1982): M o rta lity Study of Persons Exposed to Dioxin in a Trichlorophenol-Process Accident th a t Occurred in the BASF AG on November 17, 1953. An. J . Ind. Med., 3: 179-189. 28. Dalderup, L. M. and Zellen ra th , D ., (1983): Dioxin Exposure: 20 Year Follow-up. Lancet, 11: 1134-1135. 29. C arter, C. D ., Kimbrough, R. D ., L id d le , J . A ., C lin e , R. E ., Zack, M. M., and B a rth e l, W. F ., (1975): Tetrachlorodibenzodioxln: An Accidental Poisoning Episode in Horse Arenas. Science, 188: 738-740. 30. Interim Report of the Missouri D ioxin Task Force, June 1, 1983. 31. Donnell, H. D., and Hoffman, R. E ., (1983): Findings o f a P ilo t Epidemiological Study o f Missouri Residents. In a le t t e r from the Missouri governor's o ffic e to state physicians. 32. Webb, K ., Ayres, S ., S la vin , R., Knutsen, A ., Roodman, R ., Gedney, W. B ., Schramm, W., Hotchkiss, R. L ., M ille r , R ., and Donnell, H. D ., (1984): Results of a P ilo t Study o f Health E ffe c ts Due to 2,3,7,8-Tetrachlorodibenzodioxin Contamination -- M issouri. J . Am. Med. A s., 251: 1139-1140. 33. Pazderova-Vejlupkova, J . , Nemcova, M., Pickova, J . , Jira se k, L ., Lukas, E ., (1981): The Development and Prognosis o f Chronic In to x ic a tio n by Tetrachlorodibenzo-p-dioxin 1n Men. Arch. Environ. Health, 36: 5-11. 34. B leibe rg, J . , Wallen, H ., Brodkin, R ., Applebaum, I . , (1964): In d u s tr ia lly Acquired Prophyria. Arch. Dermatol., 89: 793-797. 35. O tt, M. G., Holder, B. 8. , and Olson, B. 0 . , (1980): A M o rta lity A nalysis of Employees Engaged 1n the Manufacture o f 2 ,4 ,5 -T rfchlorophenoxyacetic A cid . 0. Occup. Med., 22: 47-50. 36. Hajdu, S. I . , (1981): S oft Tissue Sarcomas: C la s s ific a tio n and Natural H is to ry , Ca Cancer J. C11n., 31: 271-280. 37. H a rd e ll, L ., Sandstrom, A ., (1979): Case-Control Study: Soft Tissue Sarcomas and Exposure to Phenoxyacetlc Acids or Chlorophenol. Br. J. Cancer, 39: 711-717. 38. Hobson, L. B ., (1983): Epidemiology of S oft-T issue Sarcoma and Related Human Research, Veterans A d m in is tra tio n , March. * 39. Cole, P ., (1980): Testimony before the Env1ronmental P rotection Agency (U .S.) In re: The Dow Chemical Company, et,. aK CCI35C5 P L S ) l? * Q q -68- 40. Rubin, P ., ed., and Bakemeir, R. F ., Assoc, e d ., (1978): Soft Tissue Sarcoma, W. B. Patterson, In : C lin ic a l Oncology fo r Medical Students and Physicians. Chapter XIX, American Cancer Society, pp. 210-217. 41. Smith, A. H ., Fisher, D. 0 ., G ile s , H. J . , and Pierce, N ., (1982): The New Zealand Soft Tissue Sarcoma Case-Control Study: Interview Findings Concerning Phenoxyacetic Acid Exposure, presentation before the Third In te rn a tio n a l Symposium on C hlorinated D ioxins. 42. Mil ham, S ., (1982): H erbicides, Occupation, and Cancer. Lancet,: 1464-1465. 43. R iih im a ki, V ., Asp, S ., and Hemberg, S ., (1982): M o rta lity of 2,4-Dichlorophenoxyacetic Acid and 2,4,5-Trichlorophenoxyacetic Acid Herbicide A pplicators in Finland: F ir s t Report of an Ongoing Prospective Cohort Study. Scand. J . Work E nviron. Health, 8: 37-42 44. Evaluation of Soft and Connective Tissue Cancer M o rta lity Rates fo r Midland and Other Selected Michigan Counties Con^ared N a tio n a lly and Statewide, Michigan Department o f P ublic Health, May 4, 1983. 45. Eklund, G ., (1983): Does Occupational Exposure to Chemical Pesticides Increase the Cancer Risk?, proceedings of Weed and Plant P rotection Conferences, Swedish U n iv e rs ity o f A g ric u ltu ra l Sciences: 6-12. 46*. P roject Ranch Hand I I : An Epidemiological In v e s tig a tio n .o f Health E ffe c ts in A1r Force Personnel Following Exposure to H erbicides. U.S. A ir Force, June 30, 1983. 47. P roject Ranch Hand I I : An Epidemiological In v e s tig a tio n o f Health E ffe c ts in A1r Force Personnel Following Exposure to H erbicides. U.S. A1r Force, February 24, 1984. 48. F lic k e r, M. R ., and Young, A. L ., (1983): Evaluation of Veterans fo r Agent Orange Exposure, in proceedings o f the American Chemical S ociety, 186th national meeting, 2: 152-154. 49. W1tt, J . M., (1980): A Discussion o f the Suspension o f 2,4,5-T and the EPA Alsea I I Study. In : Supplement to the 34th annual meeting of the Northeastern. Weed Science S o ciety, January 8 , 34: 9-25. 50. Sheldonv L. W., W1tt, J . M., Logan, A. N ., H iggins, J . E ., A g re s tl, A ., and O rtiz , M., (1979): A S c ie n tific C ritiq u e o f the EPA Alsea I I Stuty and Report, Oregon State U n iv e rs ity . 51. M a rtin , J . C. (1983): Report on OIG Review and In q u iry In to the Five Rivers In c id e n t. Environmental P rotection Agency, November 22. 4829 0013506 -69- U M SI2KM 52. Report of the C onsultative Council on Congenital Abnorm alities 1n the Yarram D is t r ic t , Commission of P ublic Health (A u s tra lia ). 53. Nelson, C. J . , Hoi son, J . F ., Green, H. C ., and Gaylor, D. W., (1979): Retrospective Study o f the R elationship Between A g ri c u ltu ra l Use of 2,4,5-T and C le ft Palate Occurrence in Arkansas. T eratology, 19: 377-384. 54. Honchar, P ., (1982): Long Island R ailroad In v e s tig a tio n , National In s titu te f o r Occupational Safety and H ealth. 55. Thomas, H. F ., (1980): 2,4,5-T Use and Congenital Malformation Rates in Hungary. Lancet, 1: 214-215. 56. Smith, A. H ., Fisher, D. 0 ., P ierce, N., and Chapman, C. J . , (1982): Congenital Defects and M iscarriages Among New Zealand 2,4,5-T Sprayers. Arch. Environ. H ealth, 37: 197-200. 57. Evaluation of Congenital Malformation Rates fo r Midland and Other Selected Michigan Counties Compared N a tio n a lly and Statewide, the Michigan Department of P ublic H ealth, May 4, 1983. 58. Townsend, J . C ., Bodner, K. M., Van Peenen, P .D .F., Olson, R. D ., and Cook, R. R ., (1982): Survey of Reproductive Events o f Wives o f Employees Exposed to C hlorinated D io xin s. Am. J . E pldeeriol., 115: 695-713. 59. Cook, R. R. and Bodner, K. M., (1983): D1ox1n and Reproductive E ffe c ts . In : Hunan and Environmental Risks o f C hlorinated D ioxins and Related Canpounds, e d ., Tucker, R. E. e t. aU Plenum Publishing Corporation. 60. O tt, M. G ., (1983): M o rta lity Among Men 1n a Chemical Manufacturing Company D is s e rta tio n , U n iv e rs ity o f Michigan, Lansing, Michigan. 61. Lamb, J . C ., Moore, J . A ., Marks, T. A ., (1980): Evaluation of 2,4-D1chlorophenoxyacet1c Acid (2 ,4-D ), 2,4,5-Trichlorophenoxya c e tlc Acid (2 ,4 ,5 -T ), and 2,3,7,8-Tetrachlorod1benzo-p-d1oxin (TCDD) T o x ic ity 1n C57BL Mice, National Toxicology Program. 62. Chesney, M. A ., (1983): Presentation to the Science and Tech nology Committee Subcommittee on Natural Resources, A g ric u ltu re Research and Environment, U.S. House o f R epresentatives, J u ly 28. 63. Case Control Study o f Congenital A bnorm alities and Vietnam Service (B irth Defects Study), M in istry o f Veterans1 A ffa irs (A u s tra lia ), January 1983. f 64. Jessop, D. S. e t. a l . , (1982): P esticides and the Health o f A u stra lia n VletnamTeterans: F ir s t Report o f the Senate Standing Committee on Science and the Environment (A u s tra lia ), November. 0013507 483,0 C D -h iq s * -70- Dow 154577 65. "Study of 85,000 Veterans C alls P esticide E ffe c t Vague," The New York Tines, August 30, 1983. 66. Moore, F ., (1980): Expected M o rta lity fo r a Population the Size and Age D is trib u tio n of Vietnam Theater Veterans 1970 Through 1979. The Dow Chemical Company. 67. Human Health and T o x ic ity Workshop on Dioxins in the Environment: Summary, U n ive rsity of Michigan, December 9, 1983. 68. "S c ie n tis ts Say D ioxin 'Not a High P r i o r i t y ', " UPI Domestic Newswi re, December 10, 1983, Update: 10. 69. Young, A. L ., Calcagni, J . A ., Thai ken, C. E ., Tremblay, J . W., (1978): The T o x ic ity , Environmental Fate, and Human Risk of Herbicide Orange and I t s Associated D ioxin. United States A1 r Force, report 0EHL TR-78-92. 70. Klmmig, J . , S chultz, K. H ., (1957): Chlorinated Aromatic C yclic Ether as Cause of So-Called Chloracne. Naturwissenschaften, 44: 337-338. 71. Schwetz, B. A ., e t. a l . , (1973): Toxicology of C hlorinated Dibenzo-p-dioxins. Imriron. Health Perspect., 5: 87. 72. McConnel, E. E ., e t. a l . , (1978): The Comparative T o x ic ity of C hlorinated D1benzo-p^3iox1ns in Mice and Guinea P igs. Toxicol. Appl. Pharmacol., 44: 335. 73. Olson, J . R ., e t. al_., (1980): T o x ic ity o f 2 ,3 ,7 ,8 -T e tra c h lo ro d1benzo-p-d1oxTn to the Golden Syrian Hamster. Toxicol. Appl. Pharmacol., 55: 67. 74. Henck, J . W., e t. a K , (1981): 2,3,7,8-Tetrachlorodlbenzop-d1o xin : Acute Oral T o x ic ity 1n Hamsters. Toxicol. Appl. Pharmacol 59: 405. 75. A lle n , J . R ., e t. a l . , (1977): Reproductive Dysfunction 1n Non-human PrimatesTxposed to D io xin s. Toxicol. Appl. Pharmacol., 41: 177. 76. A lle n , J . R ., e t. 1_., (1977): Morphological Changes 1n Monkeys Consuming a Diet Containing Low Levels o f 2 ,3 ,7 ,8 -T e tra c h lo ro d1benzo-p-d1oxin. Food Cosmet. Toxicol., 15: 401. 77. Schantz, S. L ., (1979): T oxicolog ica l E ffe c ts Produced 1n Non-human Primates C h ro nically Exposed to 50 Parts per T r i ll i o n o f 2 ,3 ,7 ,8 Tetrachlorodibenzo-p-dioxin (TCDO). Toxicol. Appl. Pharmacol., 48: Al80. 78. Murray, F. J . , e t. a l. , (1979): Three Generation Reproduction Study o f Rats GTven"T ,3 ,7 ,8 -T e tra chloro dibe nzo-p-d1ox1n (TCDO) 1n the D ie t. Toxicol. Appl. Pharmacol., 50: 241. 4^ c %.$ji 0013508 isosizaao -71- 79. Green, S ., Moreland, F. S ., (1975): Cytogenic Evaluation of Several Dioxins in the Rat. T o x ic o l. Appl. Pharmacol., 33: 161. 80. Green, S ., et_. al_., (1977): Cytogenic E ffe cts o f 2 ,3 ,7 ,8 -T e tra chlorodlbenzo-p-dioxin (TCDD) on Rat Bone Marrow C e lls . FDA By-Lines, 6: 292. 81. Wassom, J . S ., et_. al_., (1977-78): A Review o f the Genetic T o xi cology of Chlorinated Dibenzo-p-d1oxins. H utat. Res., 47: 141. 82. Khera, K. S ., Ruddlck, J . A ., (1973): Polychlorod1benzo-p-d1oxins: Perinatal E ffe cts and the Dominant Lethal Test 1n W lstar R ats. In : Chlorodlox1 ns -- O rigin and Fate, Advances 1n Chemistry S erie s, No. 120 (ed. B la ir ) , American Chemical S ociety, Washington, D.C. 83. Beatty, P. W., e t. a l . , (1975): E ffe cts o f 2 ,3 ,7 ,8 -T e tra c h lo ro dibenzo-p-d1oxin (TCCb) on Mammalian C ells 1n Tissue C u ltu re . T o xico l. Appl. Pharmacol., 31: 309. 84. Tenchinl, M. L ., e t. a h , (1977): Approaches to Examination o f Genetic Damage A fte r a Major Hazard 1n Chemical In d u stry: P re lim i nary Cytogenetic Findings on TCDD-Exposed Subjects A fte r Seveso Accident, Special Project o f In ve stig a tio n s on TDCC-Exposed Preg nancies (P ro f. G. B. Candini and P ro f. L. D eC arll), U n iv e rs ity of Milan, Ita ly . 85. Rehder, H. MeMt*. a l . , (1978): Schmelz. Ned. Mochenschr., 108:1617. 86. Kociba, R. J . , e t. a h , (1978): Results o f a Two Year Chronic T o x ic ity and Oncogenicity Study o f 2,3,7,8-Tetrachlorod1benzo-p-d1oxin (TCDD) 1n Rats. T o x ic o l. A ppl. Pharmacol., 46: 279. 87. National Cancer In s t it u te , (1980): Bioassay o f 2 ,3 ,7 ,8 -T e tra chlorodibenzo-p-sioz1n (TCDD) 1n Rats. DHHS P ublic No. NIH80-1765. 88. Toth, K ., e t. a l. , (1979): Carcinogenicity Testing o f the Herbicide TTh^-Trlchiorophenoxy Ethanol Containing Dioxin or o f Pure D1ox1n 1n Swiss Mice. Nature, 278: 548. 89. Polger, H ., and S c la tte r, C ., (1980): Influence o f Solvents and Adsorbents on Dermal and In te s tin a l Absorption o f TCDD. Food Cosmet. T o x ic o l., 18: 477-481. 90. Kimbrough, R. D ., F alk, H ., S tehr, P ., and F rie s , G ., (1983): Health Im p lica tio n s o f 2 ,3 ,7 ,8-Tetrach1orod1benzod1ox1n (TCDD) Contamination o f R esidential S o il, U.S. Centers f o r Disease Control 1n A tla n ta . 91. Health-Risk Estimates fo r 2,3 ,7,8-T etrachlorodlbenzodloxln 1n S o il. M o rb id ity and M o rta lity Meekly Report, January 27, 1984. 92. Stevens, K ., (1981): Agent Orange: A Q u a n tita tiv e P erspective. Hum. T o x ic o l. , 1: 31-39. 93. Cordle, F ., (1981): The Use o f Epidemiology 1n the Regulation o f r \ u a - c / D ioxins 1n the Food Supply. Regui. T o x ic o l. Pharmaco., 1: 379-387. 4832 001350E 4833 DON 2 16 0 2 3 7 0011537 '.i1;* ,s 'N:^v-.*r^- im. r Vi V.# <V** K -iLLli-tlO O ) TOXICOLOGYANDAPPUEOPHARMACOLOGY73, 42-47 (1984) Increased Epiderm al T ran sg lu tam in ase A ctivity follow ing 2 .3 ,7 ,8 Tetrachlorodibenzo-p-dioxin: In V iv o and in V itro Studies w ith M o u se Skin DQM2 160211 S. M . Pu h v e l , ' D . C . Er t l , a n d C a . L y n b e r g Division o f Dermatology. Department o f M ediane. School o f M ediane. Centeroe Health Sjiencrs. Univerzity o f California. Los Angeles. Los Angeles. California 90024 Received Jane 18. I98J: accepted October 25. I98J Inam ed Epidermal Tiansslutammare Activity fallowing 2J.7.(-Te!iachiorodibenzo-pdioxin: In Vivo and in Vitro Studies with Mouse Skin. Puhvel, S. M.. Er tu D. C , andLyhberg. C. A. (19(4). Toxicol. Appl. Pharmacol. 73, 42-47. In pervious audies it has been shown that topical treatment ofhxirle* mice with 2J.7,(-etrschlorDdibenro-p*ljo!un (TCDD. dioxin) induces hyperproiiferaiion and hyperkeratinixation in the epidermis of hairless mice. The present invesugation demonstrated that such TCDD-induced morphological rhsngri in skin in vivo are accompanied by increased levels in activity of epidermal tmnsflutaminase (ETG). the tn iymc mofiated with terminal epidermal differentiation. Exposure of mouse epidermal cells in tmuc culture to 10"* mTCDD abo resulted in a significant increase in ETG activity, despite the fed that morphologically these cultures (pown at 0.07 n u ionic calcium concentrations) exhibited nosignsofterminaldiflerentiation. Thusone mechanism ofaction oTTCDD in inducingcutaneous changes appears to relate to the stimulation of increased ETG levels. In humans one o f the first signs o f expo sure to 2,3,7,g-tetrachlorodibenzo-p-<iioxin (TC D D ) is the development o f a skin con dition known as chloncne (M oore, 1978). This syndrome involves hyperkeratinization o f ductal epithelium o f cutaneous sebaceous follicles, resulting in comedo form ation which may or may not proceed to pustular and cystic chloracne. Experimental chloracne has been induced in skin o f hairless m ice by topical application o f TC D D (Puhvel e t a l.. 1982; Knutson and Poland, 1982). Even though fo l licular involvem ent in mouse skin is insig nificant, the hyperproliferative, hyperkeratinizing changes induced in in te rfo llic u la r epi dermis are sufficiently well defined to perm it the use o f this animal model to m on itor TCDD-inducpd enzymatic changes in skin. In this report we investigated the effect o f TC D D on epidermal transglutaminase (ETG ) a ctivity first in skin o f hairless m ice in v iv o follow ing topical application o fTC D D . Then, to verify our observations in a m ore defined system, changes in ETG a ctivity o f mouse epi derm al cells in tissue cultures were m onitored follow ing addition o fT C D D to culture media. METHODS Chemicals. 2,3.7,(-Tetrachk>fodibenzo-p<lioxin was obtained from K.OR, Incorporated (Boston, M as.), and putrescine dihydrochloride. 2J-*H(N ) (sp. act. 3S.9 Ci/ mmol) wasobtained from New England Nudcar (Bonon. Mass). Tissueculture Medium 199, calcium depleted Me dium 199, fetal bovine serum (FBSL and antibiotic-an timycotic mixture were obtained from Grand Island Bio logical Co. (Grand bland. N.Y.k puuesdne dihydrochlo ride. carein. dithiothreiiol (DTT), and Trizma base were from Sigma Chemicals (St. Louis, Mo.); dimethylcasein was from Accurate Chemical and Scientific (Hicksvilie. N.Y.). In I'ivu Studies Animals. HRS/J strain of hairless female mice were 1To whom requests for reprints should be addressed. obtained from the Jackson Laboratories (Bar Harbor. 1> C0Alw0lif4mfI4tpa*0i8fXml/m*SSi<4iVi,SmA3an.0a0ram*yfImmtmlamcm* 42 4835' 0011538 INCREASED EPIDERMAL TRANSGLUTAMINASE ACTIVITY FOLLOWING TCDD 43 Maine). Animate wen 10to 11 weeksold atthe beginning o f theexperiment. They wereled Wayne Lablox (Universal r eeds. Inc.. Colton. Calif.) labontory chow ad libitum and were housed four to a cage in disposable plastic cages made a Biohazard safetycontainment bos (Class III Glove box) for the duration of the experiments. Treatment. Three times a week for 4 weeks animals were treated topically with 0.1 ng of TCDD dissolved in 0.1 ml o f acetone. Control animals received 0.1 ml of acetonealone. Groupsoffour mice were killed bycervical dislocation at 7. 14, 21. and 28 days after the beginning of treatment Skins were removed by dissection, and the epidermis was separated from dermis by lint immersing the dans in a 55*C waterbath. followed by immersion in ice water. Following this treatment the epidermis was soaped from thedermis with a scalpel, while maintaining the dans at 4*C. Epidermal samples were homogenized with a Pulyuon homogenize?. and the tissue cytoeol was separated by centrifugation at 30,000/ for 30 min. Su pernatant fractions were used for the tramglutaminase Transghaaminase assay o fepidermal homogenates. The methodsdesrrihed by De Youngand Battaren (1982)were used. Briefly. 0.1 ml of tim e supernatant fractions was added to 0.3 ml of assay mixture containing 0.03 m Tris buffer (pH 8.1), 0.01 m CaO), 0.03 m DTT. 0.6 mg dimedtyicasem, and 0.37 *G of CHJputrearine. Mixtures were incubated at 37*C for 60 min, and the reaction was supped byadding0.6 ml of 10* trichloroaceticadd (TCA) and 5 ml of5%TCA containing0.I * unlabeled putresrine. After 30 min at room temperature, incubation mixtures wereflhered on 23-mm Whatman GF/A filter discs. Discs were washed once with 10 ml 3% TCA containing 0.1% anixbcied putresrine and twice with 10 ml 100% ethanol. After drying, the discs were placed in Aquasol II (New England Nuclear) and the radioactivity wascounted. Sol uble proteins io the supernatant fractions were quantified by the Bio-Rad protein assaywith bovineserum albumin as standard (Bradford. 1976). /it Vitro Studies Tissue culture Basal keratmocytcculturesfrom neonatal BALB/c mice wereestablished following the methodsde scribed by Marcelo rr al. (1978). Basal ketatinocytes were separated by discontinuous neoll gradientcentrifugation, and 1.5 x 10* cells were moculatrd per 30-mm-diamcter culture dish. Cultures were grown m parallel m the fol lowingmedia: one-third oftheeeUswereplated in Medium 199 supplemented with antibiotics and 10% FBS; the re mainder were plated in calcium-depleted Medium 199 supplemented with antibioticsand 10%FBS(3%belated. 3% normal). The final ionic concentrations ofcalcium in these media were 1.2 and 0.07 mat. respectively. TCDD treatment. On the third day of culture. TCDD dmoired in dimethybulfoxide (DMSO) was added in a final concentration of 10** m to half the low calcium cultures. The remainder of the low calcium cultures re ceived DMSO alone in a final concentration of0.1%. The medium was changed every other day and fresh TCDD and DMSO were added at every medium change. Cells were harvested from four dishes of each of the three media on Days 3. 3. 7. 10. and 12 by scraping the plates with rubber spatula and collecting all cells (i.e.. attached as well as free floating), by centrifugation. Cell pellets were washed twice in buffer and stored frozen at -60*C until used for the transglutaminase assay. Transglutaminase assay o f cell pellets. Cdl lysates ob tained by thawing and freezing cell pellets three to four times were used for the transglutaminase away recom mended by S. H. Yuspa and U. Lichti (personal com munication). This procedure was an adaptsDon of the method described by Ogawa and Goldsmith (1976). Cells were pelleted in a buffer containing 30 mu Tris (pH 7.5). 2.5 him DTT. 0.13 m N ad. 0.83 m il EOTA. and 8.3 mu CaCI>. The reaction mixture consisted of 100 *1 of cell lysate. 20 *1 of casein (20 mg/ml). 10 >1 of (1H)putrearine (5 pCi), and 20 pi of buffer. The mixture was incubated at 37*C for 10 min. Following incubation 30 pi of the mixtures was spotted on filter paper discs (Whatman 3M M ) and immediately precipitated by im mersion in 10% ice cold trichloroacetic acid (TCA) plus 0.1% putresrine. The precipitates were washed in four separate 30-min washesof 3% TCA plus0.1% putresrine. then immened in 100% ethanol, and dried. The radio activity of the precipitates on the filter paper discs was countedin a Brrkman LS-7300 liquidsrintiDarioccounter. Results were corrected for backpound radioactivity found in lysate-free parallel assays. Soluble proteins in the cell lysates were determined by the Bio-Rad protein assay. To monitor the effect of 10** M TCDD on epidermal eel) proliferation, cell counts were performed at regular intervals in cultures grown in low calcium medium, with or without the pretence of TCDD in the medium. The effect of TCDD on terminal differentiation of epidermal cells in culture was investigated by transferring cultures grown in low calcium medium (with and without the presence of I0~* m TCDD) to high calcium medium on theseventhday ofculture, and mooitonngthe morphology of the cultures and the rate of comified cell envelope formation at 24 hr to 5 days after the change to the 1.2 mar CaJ* medium. Comified envelope counts were per formed according to the methods described by Yuspa rr al. (1982) with insolubility in 2% SDS and 20 mu DTT at 90*C for 5 min as the criterion for defining comified envelopes. RESULTS In Vivo Studies Hairless HRS/J mice treated topically with TCDD developed the same pattern of welldefined cutaneous changes which have been described in detail elsewhere (Puhvei et al.. r v 0011539 mooiZMod PUHVEL. ERTL AND LYNBERC 1982). Essentially, epidermal hyperplasia ac companied by involution ofsebaceous glands, hyperkeratinization of the epidermis, and keratinization of dermal cysts are the hallmarks of hairless mouse "chloracne." These gross m orphologic changes were ac companied by significant elevations in epi derm al transglutaminase activity. By the end o f the second week o ftreatment, the epidermal transglutaminase (ETG) levels were m ote than sixfold the baseline levels, o r the levels in the skin o f acetone-treated mice (Fig. I ). By the end o f the th ird week o ftreatm ent, ETG levels had decreased from the in itia l peak but s till remained at more than twice the baseline levels at the term ination o f the experim ent at 4 weeks. To test for the possibility that TCDD pres ent in the treated mouse skinswasearned into Mens o r TWATHtMT Fig. 1. Epidermal mMShitsminaae activity in skin of hairteB mice treated with TCDO ( ------ ) and in con trols (O -- -O k esptoaed as cpm incorporated duiias 30 min of incubation/mg of epidermal cytoaoi protein. Sec Methods far details. the ETG assay and exerted a direct activating effect on the enzyme in the epidermal cytosol preparation, 0.2 ng o f TCDO was added di rectly to the ETG assay mixture and the effect monitored. No changes in radioactivity of the resulting precipitates was observed. In VitroStudies t. The ionic concentration o f calcium in th ^ medium has been well established as a reg u -^ lator o f mouse epidermal cell terminal d iffe r-^ entiation in in vitro tissues cultures (Hennings0 3 et at., 1980) and our results were in line w ith o this finding. In low calcium medium, mousee keratinocytes did not undergo terminal dif- 4 W ferentiation but continued to proliferate u A basal cells in monolayers (Fig. 2A). Addition o f 10~* M TCDD did not change this pattern o f growth at least in so far as could be deter mined by light microscopy (Fig. 2B). Nor did addition o fTCDD have any effect on the rate o f cell proliferation as measured by compar ison o f total cell counts o f cultures, 24, 48, 72, and 96 hr after the addition of TC D D or DMSO to the low calcium medium. Total protein content (measured to the twelfth day) was similar in cultures grown with or without TCDD. Cells grown in high calcium medium started to stratify and differentiate 72 hr after plating (Fig. 2Q . ETG activity in the TCDD-treated ceils in low calcium medium gradually increased over the 12-day period of growth, so that on the twelfth day ETG levels in the TCDD-treated, undifferentiated cells were almost identical to levels in the fully differentiating cells grown in high calcium medium (Fig. 3). ETG levels in the undifferentiating cellsgrown in low cal cium medium without the addition o fTC D D also increased slightly over the 12-day incu bation period, but remained at about half the level of activity in the previously described cultures. That this was a genuine increase of ETG production by the cells, rather than an acti vation o f ETG activity already present in the 13cells, wasshown by the fk t that 10~*m TCDD added directly to the transglutaminase assay did not have any effect on the results. 0011540 INCREASED EPIDERMAL TRANSGLUTAMINASE ACTIVITY FOLLOWING TCDD 45 treated cells (0.7%) than in the control cells (0.3%), but it remained much lower than the counts of comified envelopes in cells grown in high Ca2* medium from the beginning (12 to 14%). Morphologically, terminal differen tiation and cell death were delayed in the TCDD-treated cultures, in a manner similar to what has been reported for retinoic addtreated cultures by Yuspa et al. (1981) (Fig. 4). DISCUSSION The changes induced in human skin by TCDD exposure are among the more observ able and well-defined biological effects of this potent man-made toxin. In the hairless mouse model, topical application o fminute amounts ofTCDD induces a distinct hyperpreiiferative, hyperkeratinizing response(Puhvel etal. 1982; Knutson and Poland, 1982). Epidermal trans glutaminase is considered a marker enzyme for terminal epidermal differentiation (Gold smith and Martin, 1975; Buxtnan and Wuepper, 1976). This soluble, calcium-dependent enzyme which catalyses the formation o f co- I f D0H2I602M \ Fie. 2. Neonatal mousc epidemial ee> cultures (A) in 0.07 k m Cm1' mdium; (B) in 0.07 m Ca2* and 10** m TCDD; and (O in IJ m Ca2" mdium al 7 days after piaunj. Note ihai TCDD did a afcet cell culture mocphology in the low calcium mdium. Changing kentinocyte cultures from low calcium to high calcium medium on Day 7, at a time when it had been established that ETG activity was high in the TCDD-treated, undifferentiated cells, indicated that termihal differentiation was not the same in TCDre treated cells as in control cells grown in low ilcium medium without the presence of CDD. The comified envelope count was consistently slightly higher in the TCDD- *rnto MM Fig. 3. Epidermal irensfjutaminaie activity in cultures frown in 1.2 rim Ca2* ( ------ ) . in cultures frown in 0.07 him Ca1* plus Iff* m TCDD (O ------OL and in 0.07 r im Ca2* without TCDD (A ------A). 4838 o n 34 0011511  ItZ O S U M O Q 46 fUHVEL. ERTL. A N D LYNBERG FC-4.ffluDO>oitiedekyiannniMl diWertnoMion ofwon* rpitfcmnlrdlculnim foUowiaetremacntwith KT*m TCDD. PmUdcultura werefan grownin 0.07 M Of* for 7dnjnin theprwenct (A) andwithoutthe practiceof I0~*MTCDD(B).Culturelwerethenwitched to 1.2w nO f*(withoutTCDD).Tcnnianldiflbamarion andedidth wondelayedintheTCDDmcniodcultuia (A)comparedtocontrols(B) Thephowgreph>n taken 5day*aftertremfer to U am Of* valent crosslinks, involving(y-gluytamyl-lysine)dipeptide bonds, isthought to beinvolved in the conversion of soluble structural protein beneath the plasma membranes of epidermal cells, to insoluble, high-molecular-weight pro teins which form the comihed cell envelopes of differentiated keratinocytes. Levels of transglutaminase activity in vivo in mouse ear epidermis are affected by agents which alter epidermal differentiation, such as anthralin, retinoic add, and fluocinolone acetonide (DeYoung and Ballaron, 1982). In the present in vivo studies, a significant increase o f this enzytr& activity wasdemonstrated fol lowing changes in cutaneous differentiation after TCDD application. One possible com plication in interpreting results of in vivo ex periments which -involve analyses of crude epidermal extracts is that during the dermalepidermal separation procedure, dermal transglutaminases may theoretically contam inate the epidermal preparations. Substances applied topically to the skin, particularly those which induce an inflammatory cutaneous re sponse, may affect the dermal as well as epi dermal transglutaminase levels. Previous studiesbyDeYoungand Ballaron ( 1982)which investigated this problem suggested that their results were not affected by inflammatory changes. In the present paper, in vitro tissue culture studies were used to confirm the in vivo observations in a system which was free of dermal components. The'observation that mouse epidermal ker atinocytes in low Ca1* medium have lower levels of transglutaminase activity than dif ferentiating keratinocytes in high Ca2* me dium has been established previously by Hen nings et al. (1981). Our finding that TCDD increases unexpressed transglutaminase activ ity in the nondifferentiating cells in low Ca2* medium confirms our in vivo observations about the increase of this enzyme by TCDD in epidermal cells. Very similar effectson transglutaminase in duction by epidermal basal cells in vitro have been demonstrated by the addition o f retinoic add (Yuspa et al.. 1981) or 12-0-tetradecanoylphorbol-13-acetate and other tumor pro motors (Yuspa et al.. 1980) to low caldum mouse epidermal tissue cultures. In those studies as in the present experiments, it ap peared that the effect was one of induction ratherthan activation ofenzyme activity, since addition of the chemical (in this case 10"* M TCDD) directly to the enzyme assay had no effect on the results. In the studies by Yuspa et al. (1980,1981 ) retinoic add had a different effect on terminal differentiation o f the cell culturesafter transferto highcaldum (1.2 m M ) medium. Normally, when epidermal basal cells are transferred to high caldum medium after bang cultured in low caldum medium for 5 to 7 days, terminal differentiation ac companied by cell death occurs within 2 to 5 days after transfer. In cells grown in the pres ence ofretinoic add (at 10"* M concentration) 00115 INCREASED EPIDERMAL TRANSGLUTAMINASE AC TIVITY FOLLOWING TCDD 47 this pattern of terminal differentiation was significantly delayed. Phorbol esters did not affect the rate of terminal differentiation. In the present study, TCDD also delayed ter minal differentiation of the cultures, despite the fact that insoluble cell envelope counts appeared to be higher in the TCDD-treated transferred cultures than in the untreated con trols. Knutson and Poland (1982) have suggested that cutaneous sensitivity to TCDD in mice is associated with the gene for faairiessness, and that hr/hr (hairless) animals have a more acute cutaneous response to TCDD than do the hr/+ (haired) counterparts. In the present studies, epidermal cell cultures were estab lished with BALB/c hr/+ (haired) neonatal mice as tissue donors. Interestingly, such cells in culture responded to TCDD exposure. It "ould be that the effects o f TCDD on epider- tal cell cultures would be even more striking in epidermal cultures from hr/hr (hairless)skin donors. ACKNOWLEDGMENTS Thn fc w c h w it supported in pan by G n a ii AM 17423 and OH 1108 from the National Institute ofHealth, by VeteranAdministration Medical Researchfunds, and funds from die Dermatologic Research Foundation of California, Inc REFERENCES Bradford, M. (1976). A rapid and sensitive method for the quantitation ofmicrof/am quantitiesofprotein uti lizing the principle of protein dying binding. Anal. Biochem. 7 2 ,24S-234. Buxman. M. M.. and Wuepper. K. D.(I9761 isolation, purification and characterization of epidermal trans glutaminase. Biochim. Biophys. A aa 452, 336-369. DeYoung, L , and Ballaron, S. (19823. The effect of topical drop on mouse ear transglutaminase activity. J. lim a . D erm atol 79, 189--193. Goldsmith. L. a , and Martin. C M. (1973). Human epidermal Isansaraidase. J. Im est. Dermatol 64,316321. Hennings, R , Michael. D.. Cheng. G . Steinert. PHolrrook, K_ and Yuspa. & H. (1980). Calcium regulation of growth and differentiation of mouse epi dermal edit m culture. Cell 19,243-234. Hennings, R , Steinert, p,, and Buxman. J*. M. (1981). Cairium induction oftiansglutaminur and the formation of<(y glutamyl) lysine crow links in cultured mourn epidermal edit. Biochem. Biophys. Res. Comm m . 102, 739-743. Knutson, J. C , and Poland, a . (1982). Response of murine epidermis to 2,3,7.8-tetraehlorodibenzo-ptboxin. Interaction of the Ah and hr too. Crtf 30,223234. Makcelo, G . Kim. Y. G,, Kleine. j . 1_ and Vorhees. J. J. (1978). Stratification, vernalization and prolifer ation ofprimary fcoatinocyte cultures. J. Cell Biol. 79, 336-370. Moore. J. A. (1971). Toxicity of 2J.7J-tetiadilorodibenzo-p-dioxia. taf. BoU (Stockholm) 27, 134-144. Ogawa, R , and Goldsmith. L. a . (1976). Human cpidermal transglutaminasr J. Biol. Chem. 251. 728172U. Plihvel, S. M - Sakamoto, M ,, Ertl, D. C . andRejsnejl R. M. (1982). Hairtes mice as models for dtioraene: a study of cutsmoot changesinduced by topical application o fcitabfishndddomcacgens. Toxicol. Appt. Pharmacol 64.492-503. Yuspa, S. R , Ben. T ,, Hennings, R , and L o rn . U. (I9S0). Pbortolcsttrtumorpromotersinduceepidermal transglutaminase activity. Biochem. Biophys. Res. Common. 97. 700-701. Yuspa. S. H - L o m . U-. Ben. T_ and Hennings, H. (1981). Modulation of terminal differentiation and teiponies to tumor ptomoten by retinoids in mouse epi dermal cell cultures.-/*it*. N .Y. Acad. Sci. 359. 260273. Yuspa. & . Ben. T ,, and Steinert. P. (1982). Retinoic add induces transglutaminase activity but inhibits cornification of cultured epidermal cdls. J. Biot. Chem. 257.9906-9908. / 0011543 $1Z09U N O Q  li 11 S M "' 3 '1 M 888U KO I 2.4. I RESULTS FROM 8 6 TW O-YEAR f A. CARCINOGENICITY STUDIES CONDUCTED BY THE NATIONAL TOXICOLOGY PROGRAM OH J. K. H u m a n , D. D. Crawford od- Biometry and Risk A>tnwnen( Proram. National Jnuione of Envnonmcntal Health Scienccv Research frianflc Park, North Carolina TV 3N /. L Huff. G. A. I i , L L McConnofl Natioruf Toxicology Program, National Im titutc G. of Enwranmontal Health Sciences, Research . Triangle Park, North Carolina ./ > " * > ir m - f. </ ru a w i r */ l a i w m r a iw a vi 4 M s r < r u v a M V**** - i w w ;a .v > rw n <w f t rtem arw i iim M m v r r u r f a n --v r --i m ir v i ra m n r - v L * (Or Vn n im /M irate) Oraya-- (M fPf. O t n r v n w im , io / (.a d / o r rr- M M a i ( ( n m o M r r r n ix 44% ( M /tt) a n v -- materne#O la i w o o r v i. AH ' i l t t l 1A >n aoweeta# ru v r - r r iv r a o n o w ir r , amr f i l i t i - tr a n om a r* a i laaarrvaw n r r r m m v /O r a r m (Or w Aio k m v r r r ancrr - O r 4 /n rO rr- J44 ra n W m M K ano Im a U C J f, mar. Mar ra d aro a rra m ar w w iw Ma (InoOIr rad (a (Or moacn aw a / moQteite. o a r te r a w r (Or Iw a r i wemwr a a a m o a a Uir . /O r ra a rrt a / aOon i rr anmi n rM n f Or 0^ O rtr im m ia p r f 10-13*> t f m i iwr M n rr o m o a ra a M M a * . a M m u a a ik ' w O aV > Or f n y ariamo -a n r. a / ari-- / v /in lO a -rV con m nftaa rO rrrv te ir M -- 1 m aire vurraa vrrai a u va aoa con tro l a m a ri - r r r vm rter, aOUr Or o tte n n i. *> * Vm I iO aara nam te aariy rra o cra ornava a a n r a r amar teice am ar fru tto * m am i< -n n n a iM ( I -- m a i /O r a w ia # rrrcrw ra a r a / ttntr* tnumam ar w io a a v flirtn HOMI a w n Cbnrfy m ro (Or rara rrm a irtl n -M r r a a iin a w i te r arar/ ZOO ratear rarram arm ra r u r r r imr ar i a a M irv a (Or Vvte iote ra m a r te w xw r 0013395 INTRODUCTION Since the National Toxotcoiogy Program (NTPJ was given response bility for the National Cancer Institute (N C I) Carcinogenesis Bioassay Program in July 1981. the results of approximately 8 6 chronic studies in rodents have been published as technical reports or prepared in draft form 'H u ff et al.. 198S' H uff and Moore. 1984). The majority o f these studies are 2-yr feeding jir Ravage experiments involving groups o f SO male and ` s ic -r. r- i.eteo rW yatfc* p-4< ? 3-r T5T2" 622 I, H HASEMAN ET aL female Fischer-344/N (F344) rats and B6C3F, mice and used experimental' protocols similar to those employed by the NCI in earlier carcinogenicity studies (Sontag et al., 1976; H uff, 1982). A summary of the test result* from approximately 2 0 0 of these earlier experiments has been given by Griesemer and Cueto (1980) and by Chu et al. (1981). The purpose of this paper is to provide similar summary information for the more recent NTP studies and to examine other factors such as species and sex sensitivity route of administration differences, survival effects, and relative frequency of site-specific carcinogenicity. Chemicals have been nominated, selected, and tested for carcinogenicity by both the NCI and the NTP using criteria such as human exposure, use and level o f production, chemical structure, and available mutagenicity data (H uff, 1982). Priority for testing has often been given to those chemicals for which there is some a priori suspicion o f a toxic or carcinogenic response. However, selection per se is not and should not be taken as an indicator o f a chemical's ultimate carcinogenic potential In experimental animals or in humans. M ATERIALS AND METHODS The Griesemer and Cueto (1980) summary o f test results includes an chemicals with technical report (TR ) numbers 1-196; the summary provided by Chu et al. (1981) extends from TR 1 to 190. The evaluation in this' paper begins with TR 197 and includes all chemicals whose TRs have beta peer reviewed by the NTP Board o f Scientific Counselors* Technical Reports Review Subcommittee and Associated Panel o f Experts (Hart et aL, 1983; National Toxicology Program, 1984) as of July 1984. Approximately two-thirds o f these reports have been printed; the remaining one-third' have had their conclusions approved by the peer review process. Certaa of these chemicals are presently undergoing a data audit by the NTP, but 1 t is unlikely that these ongoing audits w ill alter the basic conclusions of the reports. In only one instance (methylene chloride; not included in this evaluation) have the audits to date revealed problems that would rendtr the studies inadequate or otherwise affect the final interpretation. The 8 6 NTP carcinogenicity studies were, divided into four groups studies regarded as showing carcinogenic effects (Table 1), studies with equivocal evidence o f carcinogenicity, i.e., studies showing some suggestion of carcinogenicity, but the strength o f the evidence was judged insufflates fo r a definitive conclusion o f carcinogenic (Table 2), studies interpreted a showing no carcinogenic effects (Table 3), and inadequate studies (Table 4). These groupings are similar to the five categories o f evidence of car cinogenicity recently adopted by the NTP (H u ff and Moore, 1984), w i* the following exception: rather than attempt to classify retrospeetmh the NTP studies considered positive regarding whether they showed "dear" or " some" evidence o f carcinogenicity, these classifications were pooM (Table 1). S9 i cn JaV f II i 1* J ii la | a ! 1 1 1 4ii I* IaI3 &i --i C' 1 ! !zy Jc Om U ^wmIUk 3 V* ?* 5 4I J 5. 5 J. K. HA5EMANETAL. ce and used experimental in earlier carcinogenicity nmary o f the test results nents has been given by (1981). The. purpose of tion for the more recent pedes and sex sensitivity, s , and relative frequency tested for carcinogenicity human exposure, use and iiabie mutagenicity data given to those chemicals or carcinogenic response. 3 t taken as an indicator experimental animals or test results includes all 5; the summary provided . The evaluation in this Is whose TRs havt been elors' Technical Reports perts (Hart et al., 1983; 1984. Approximately he remaining one-third review process. Certain audit by the NTP, but ie basic conclusions of de; not included in this ems that would render terpretation. ded into four groups: Table 1), studies with owing some suggestion was judged insufficient , studies interpreted a* equate studies (Table ies of evidence of carid Moore, 1984), with classify retrospectively ;rth e y s h g w q d "c le fications *i IA 'p - 4 5 ^ 0 . '-M;l ,o o 0 0 -- OlW to i-- urn TABLE t. NTP Studies Interpreted u Showing Carcinogenic Effects {Continued) --------- - ..... -- * .......... - ------- Rat l * l0 . Chemical (CAS no.) Boute/dose Site or type ol tumor MF 2-Mphetsylamlne HO (2103-92-4) Bls(2-ctilafo-l-meltiyl ethyl) ether (10040-1) 1,3-Buledlene (100-9941) 8 Chlorobenierie (106-90-1) ChlorodIbromomethane (124-46-1) C .I. disperse yellow 3 (2632-40-6) C.I. solvent yellow 14 (642-07-9) F344 B6C3F, B6C3F, 86C3F, F344 B6C3F, F344 B6C3F, F344 B6C3F, F344 06C3F, Feed: 1000 or 3000 PPm Gavage: 100 or 200 mg/kg Inhalation: 623 or M 1 0 ppm Gavage: 60 or 120 mg/kg (rais and Ie male rnlce): 30 or 60 mg/kg (male anice) Gavage: 40 or 60 mg/kg (rais); 30 or 100 mg/kg (m ke) Feed: 3000 or 10,0 M ppm (rata); 23M or SOM ppm (mice) Feed: 230 or 3 M ppm (rate); SM or I0 M ppm (mke) Circulatory system: hemanglo- , MfCpmM Lung: alveolar/bronchlolar adenomas Liver: carcinomas Forestomach: squafttous-cell papU- lomas or carcinomas (combined) Heart: hemanglotarcomas Malignant lymphomas Lung: alveolar/bronchlolar adenomas Lung: alveolar/bronchlolar carcinomas Stomach: papillomas Mammary gland: acinar-cell carcinomas Ovary: granulosa-ccll tumors Llver:catdnomas or adenomas (combined) Liver: ncoplaslk nodules Liver: carcinomas Liver: adenomas Liver: neoplastic nodules Liver: adenomas Stomach: all tumors Malignant lymphomas Liver: ncoplaslk nodules E Mouse MF E TA no. 233 239 E 29 t t 261 E ,3 212 222 E 226 ) P h i^ ft l.'ylembena U U S 9-9I-J| O and C rad no. 9 (3160-02-1) l,2-Dlbromo-3-chlofoptopat>e (OBCf) (96-12-1) H44 M C1F, F344 B6C3F, F344 B6C3F, Intraperitoneal M *cbm, 1 tlmes/wk: 1 oc 14 mg/kg (rali)j 12 or 24 mg/kg (mke) Feed: 1000 or 3000 ppm (rau); 1000 or 2000 ppm (mke) I unica vailnalUs mewfhcllomu Multiple organs: melotticitomai Mammary gland: fibroadenoma Spleen: sarcomas liver: neoplastic nodules Inhalation, C h/d, 5 d/wk: Nasal cavity tumors 0.6 or 3.0 ppm Tongue: squamous-cell papMomas t t 201 cn 22S 206 Chlorodlbromomoiliana (124-41-1) I i* ' i ' 'J '-M.i. ij;t|L yHow 3 (2112 401) V*Muw 14 (42-01-4) i #4 . V i (2173-1-1J ?> ! 1$1 V ' t." "1 rad no. 9 (5160412-1) tomo-3-thloroptopano (DOCP) 2 ) F344 B6C3F, F344 06C3F, Gavago: 40 or 0 m/kg (ran); 50 or 100 m/kg (mko) Food: 5000 or 10,000 ppm (ia ll); 2500 of 5000 ppm (mko) F344 04C3F, Food: 250 oi J(to ppm (fan); 500 or 1000 ppm (mko) ---------- F344 M C3F, F344 06C3F, F344 BCC3F, lolfopoflloooal bi|ooHon, 3 tlmoi/wk: 7 or I4aag/kg(rali);. 12 or 24 mg/kg (mko) Food: 1000 or 3000 ppm (ran); 1000 or 2000 ppm (mko) 0.6 or 3j0 ppm omoolbano (oihyltno dferomldo) 3- F344 B6C3F, Inhalii(on, C b/d, S d/wk: 10 of 40 ppm , ' a ` V.il`1 1*.'. * >*4#- m'ii i> `m H4%,-,'.i.ll V\i'-i-*.fl^m-i)h-lorp-plMiiyl<MdlMiliM * ! ;r ' ' * 120.1) . L 'J ! r -<o. V oo CCCCOOOO! *.hri?itoropfOfUM 't :r|F>i (7-7-S) J cn F344 06C3F, F344 B6C3F, Food: 1000 or 2000 ppm (nut* rala); 2000of 000ppm ((mal rail); 1000 of 3000 ppo (mko) Gavago: 62 of I2S m f/kf (malo ra li), I2S of ISO m g/kg (mko and fornaio ra li) Llwri carcinomas L h m mIm m u Liver: neoplasik nodules U rn : u fa iw M Stomach: all tumors Malignant lymphomas U v iti neoplasik nodules E E 262 222 E 226 " Tunica vaglnallsi mesotheliomas Mltiplo H |m : mesoihelhsnias Mammary gland: fibroadenomas Spleen: sarcomas U m : neoplasik Modulas Nasal cavity tumors Tongue: squamous-cell papMomas Tongue: squamous-call carcinomas Adraaal glands: conical adanomas Lung: etveolar/bronchlolar adenomas Nasal-cavliy tumors Circulatory syslam: hemangiosar- comas Tunica vaginalis: masolhallomas Lung: alveolar/broneMolar adanomas Lung: alveolar/bronchioles carcinomas Lung: alveolar/bronchioles adanomas or carcinomas (combinad) Subcutaneous fibrosarcomas Mammary gland: fibroadenomas Mammary gland: adenocarcinomas Liver: adenomas Liver: carcinomas or adanomas (com binad) * 207 223 206 210 21 Liver: adanomas Mammary gland: adenocarcinomas 263 E imiZMOQ TABLE I. NTT SM 4lu liM ip riliJ as Showing Carcinogenic Effects \C onlkiutd) Chemical (CAS no.) OI(2-ethylbej(yl) adipate (OEHA) (103-23-1) DI(2-elhyRicxvl) phlhalale |0EH F| (11741-7) Animal strain* F344 86C3F, F344 M C3F, Dl^ycldyl resorcinol eihr (OGRE) (101-904) F144 BSC3F, s Mm*thy! hydrogen phosphite ( I M I 5 t ) F344 R6C1F, Ethyl acrylate (I4 M F 5 ) FS44 86CSF, Rouu/doae Fn 4: I2/W 0 o t 23/WO ppm Feed: 6000 or 12/W0 ppm (rala)i 3/WO or 6,000 ppm (mice) Gavag: 12,25, or $0 mg/kg (rals); SOor 100 mg/kg (mice) Gavag: 100 or 200 mg/kg (mal (an, m k i);5 0 o r 100 mg/kg ((mala (ala) Gavag: 100 or 200 "yAi HCM m m . I (2704-94-3) F344 M C3F. F!: 1300 or 3000 ppm (rata, mala mica); 2000 or M 00 ppm ((amala mice) Rat Sll or lyp ol lumor MF Liver: adenomas Liver: carcinomas Liver: carclnomaa or neoplastic nodules (combined) Liver: carcinomas + Siomach/forctiomach: aquamoua-cell paplllomaa Slomach/loreatomach: aquamoua-cell carclnomaa Lung: alvaolar/bronchlolar adenomaa Lung: alvaolai/bronchlolar carclnomaa Lung: squamout-cell carclnomaa Forealomach: squamous-celt papll- lomaa or carclnomaa (combined) Forealomach: rquamous-ctll paplllomaa Forealomach: squamous-cell car- clnomaa Forealomach: aquamoua-cell papll lomaa or carclnomaa (combined) Liver: carclnomaa Llvar^carclnomaa or naoplaallc nodules (combined) Lung: alvcolar/bronchlolar adenomaa or carcinomas (combined) t hyroid: follicular cell adenomas + >' Mouse MF 1 : t TR' no. 212 217 2S7 207 239 271 0013400i L 4847 1leMKMere4llHiefi-4loa|ftt, 1.2.3.6.7.0-and l,2.1.7,a,9(37653-05-7X and l940-74-3) Melamin (100-70-1) OsborneB6C1F, F144 B6CJF, tUvage: U S , 2.3, or S Hg/kg-wk (tala, male mtm); > J, 3, or 10 pg/ kg-wk (female mice) Fd: 2230 or 4300 ppm (male rata, mice); 4300 or 9000 ppm (female rals) Uver: neoplaulc nodules Uver: adenomaa Liver: carcinomas Liver: carcinomas or neoplastic nodules (combined) t a E ta Urinary bladder: translllonal-cll 4 carcinomas (associated with ' bladder alonea) 692B S IZ M O fl t9B 24S F344 B6C3F, F *l: 1500 or 3000 ppm | iiu , Aula mica); 3000 or 6000 ppm (famala mica) <Mada B6C3F, Glvipa: 1.35,3.5, or 3 a*/ki*k' (fU. mal* m ka);2.5. 5, or 10 m / k f >wk ((amala mka) F344 B6C3F, F344 M C 3F, Fa*4: 32SO or 4500 ppm (mal* rail, mica); 4500 or 9000 ppm ((amai* rala) Drinking vaiar; 150 or 300 ppm F344 B4C3F, F344 M C 3F, FtaOs 750 or 1500 ppm (rau); 5000 or 10,000 ppm (mica) Faul: 300,400, or 500 ppm (rau) 150.300, or 000 ppm (mica) F344 B6C3F, Gavafa: 75 or 150 m tlk g (rau); 250 or 500 mg/k| (mica) U ! Foretlomach; tquamout-cell papiomai or carcinoma* (combinad) Llvar: carcinoma* Llvar: carcinoma* or ncoplatilc nodulat (combinad) Lung; alvaolar/brondilolar adanomu or carcinoma* (combinad) Thyroidi follicular call adenoma* E t 271 a Llvar; naoplaulc nodule* Llvar: adanomu Llvar: carcinoma* Llvar: carclnomu or naoplaulc nodulat (combinad) Urinary bladder: iraotlilonal-ceN carcinoma* (auoclaud with bladder iron**) Thyroid; fotticular-cell adenoma* Thyroid; (oHIcular-call carcinoma* TkyroMl e-cad adenoma* Llvar; naoplaulc nodulat Llvar: carcinoma* Adranal: pheochromocyiomat Malignant lympbomu Kidney; tubular-call adenoma* Kidney: lubular-call adenocar cinoma* Llvar: naoplau lc nodulat Llvar: neoptattic nodule Llvar: adenoma* Llvar: carcinoma* Llvar: adenoma* or carcinoma* (combinad) Thyroid: loMcular-caN adenoma* Thyroid: follicular-call carcinoma* Hardarlan gland: adenoma* Llvar: adenoma* Llvar: carcinoma* Kidnay: tubular-cad adanomu E a a a a aa aa aa a .a E 196 245 t 241 a a 266 205 a a a a aa a 232 aa titSfilZNftfl TABLE 1. HTP Sludlea ln M fiM l at Showing'Cardnogenk Effedt (Continued) Chemkaf (CAS no.) (Flramaatar F F -I) (07774-12-7) O' , Propylene oxMo (75-50-5) Talona II (542-75-0) Animal M ain* F144 00C1F, F144 0CC1F, F344 B6C1F, R o a li/M a Gavaga: 0 ,0 .1 ,0 .1 ,1 0 . 1.0, or 10 mg/kg Inhalation: 200 or 400ppm Gavaga: 35 or 50 mg/kg (rata); 50 or 100 mg/kg (mica) i , V > T n m liln i O iM > g Eaula (TCDD) (174641-6) H T ^ -T ilr aA lw ol lla a ta g Ew Ja (TCDD) (174441-6) 1,1,1,2-Tetrachloroalhane ( U f r lM ) Osborne B6C3F, Swlae- WtkMr F144 B6C1F, Gavage: 0.01,0.05,or 0.5 pg/kg^nk (ralt and nab mice); 0.04, 0.3 or 3.0 (lamala mica) Dermal: 0.001 >ig (mala mica); 0.005 M (faa A mica) 3 d /*k Cavagal 135 or 350 mg/kg (rati); 350 or 500 mg/kg (mica) Ral Site or lypa of tumor M Llvar: naoplank nodulaa Uver: carcinomas Uvar: cholanglocarclnomaa Naaal turblnalaa: papillary adanomaa Natal turblnalaa: hamanglomaa or hamangloaarcomaa Forartomach: squamous-cell papHlomaa Foraalomach: squamous-cell carclnomat ForcMomach: iquamout-ccll paplllomaa or cardnomaa (combined) Llvar: neoplastic nodulaa Urinary bladder: transitional-cell cardnomaa Lung: alveolar/biondilolar adanomaa Thyroid: folllcular-call adanomaa Llvar: neoplastic nodulaa Llvar: cardnomaa 4 4 4 F 4 4 4 4 4 4 Integumentary aystemi flbioiarcomaa Llvar: adanomaa Llvar: cardnomaa Mouse MF TR ' no. 244 44 267 44 1 269 4 4 44 ! 44 209 E ;4 1 201 i' 4 4 > 237 4 li' 2 1S H r i ialuene dNaucyanale (1,4- and 2 ,5 lamnata) |2647l-42-5) F144 Gavaget 50 or 00 mg/kg SidiculaiMoui llbiomaa or flbtotar- a a M C 1F, (mala rata); 00 nr 120 comaa (combined) 251 mg/kg (lamala rata); Panaaaa: aclnar-cetl adanomu a 130 or 340 mg/kg Panctaaa: Mel-cell adanomaa a I lJ (mala mica); 00 or 120 mg/kg (lamala mka) Llvar: naoplasllc nodulaa Liver: adanomaa Mammary gland: ffttroadenomaa a a a Circulatory ayalam: hamanglomaa or hamangloaarcomaa (combined) a Trichloroethylene (without epkhloro- FJ44 Gavage: 500 or 1000 Llvar: adanomaa I a 241 2,]lI ll-TUncklo(o#iiu'HM (|C D D ) (1746-01- i) t.ti^ -T tlrK M iirM lh iM (430-20-6) .. ; i i l r:m M L ii, Swiu- Wibriv F344 06C3F, IMIUUM>IM)j0.04, 0.2 or 2.0 m A<*wk ((mala mica) Dannai: 0.001 Mi (mala mica); 0.003 > (ra mala M ic i)) d/w k Cavata: 123 oc 230 n>tlk| (fata); 230 or 300 mt)ka (mka) Ulvat: carcinoma InMfumantary aytlam: Itbroiarcomu Uvar; adanonrar Uvar: cudnomu at E 201 237 immO bacyiM H (2,4- m 4 i > mu) (2647162 3) .H j- nriiloroailiylaaa (wldioiit apkhloroI '( ''^ J J 'd c ln (24-01-4) ) I4', , iM (2-ENiyllM Jiyl) pbaitbala Jjp3-42-2) <^ f ' ?JfeXyWta. (37-42-7) F344 06C3F, F344 B6C3F, F344 B6C3F, CO-I Cavaga: 3 0 o rM m t/k 3 (mala rau); M a r 120 mt/kg (ramala m i); I2 0 o r 24 0m f/k j (mala mka); M or I2 0 m /k | (lamala mka) Cavata: 300 or 1000 m t/kt (rau); 1000 m f/k| (mka) Gavata: 2000 or 4000 m iA ( (mala rau); 1000 or 2000 mc/k (ramala rau); 300 or 1000 a t|/k | (acka) Fami; 0,300,1000 or 3000 ttm Subcui anaour librom i or flbroiar- Fancram: actnar-call adanomu Fanctaui Wal-caH adanomu Uvar: naoplaatk nodulo Uvar: adanomu Mammary (land: llbroadanomu Clrculaiory tyuam; bamantlomu or baman|loucomu (comblnad) Llvar; adanomu Uvar: carcinoma Adraaal; phaochiomocylomu Llvar: cudnomu Naaal cavky: tapINary adanomu Naial cavky: cudnom u ' 1jld r a k nona |1H 14-M -4| f F144 D6CSF, Faad: 23 or 30 ppm (rau); 30 or ItO y ra i (m ka) Pliukaryi adanomu Uvar: adanomu (137-30-4) ?% F344 B6C3F, Faad; 300or600ppa (rau); M 0 or 1200 ppm (mka) Tfcyrold: c-caM cudnomu Lnnti alvoolu/bronchlolu adanomu l tiralo*: F344 (Fkdiar-344);Oibonia Mandai; CD-I (Chulas Rivai CD). Moina uralai: B6C3F, | Swbt-Wabuar. o j#F:Oburvod aliaci coruidarad equivocai; |: ttudy M f*d loadaquaie. o * a | . .^5 afe V / S * ' ' $ 00 ai o * t 231 243 274 273 233 233 t i m i no* TABLE X NTP S tuditi Interpreted u Showing Equivocai E44net of Carcinogenicity ChniIrai (CAS no.) Butyl bonayt phtealatt (IM S -7) AM O! Italo* F344 B4C3F, Roult/doat Food: 4000 or 12,000 rpm Slit oa typ* of lumoy Mononuckaa-ctN kufctanla Rat MF 1 E6 Mourn M- F TR no. __c----- ; 213 ; C.I. add yaBtw IS (nuotate*In aodlum) (S IM 1-S ) F344 BSC3F, Printing waUt: 2500 oa 5000 ppm (rata); 5000 oa 10,000 ppm (m kt) Fanertaa: ftltt-ctN adtnomaa or caaclnomaa (comblntd) E .. 245 MaMylplHhalato (131-17-3) F344 B4C3F, Gavaga: 50 oa 100 mg/kt Mononuclear-cell Ituktmla Gayagt: ISO oa 300 mg/fcg Malignant lymphoma E E 214 242 Eugtnol (97-53-0) F344 R6C3F, Food: 4000 or 12,500 ppm (ftmala iati); 3000 oa M 00 ppm (malt atta and m kt) Liver: adenoma or caaclnomaa (comblntd) E E 221' Sodhim (2-tlhyBMxyl) auKalt (I26-S2-J) F344 B4CJF, F ttd: 10,000 or 20,000 ppm (rata and funai* m kt); 5000 oa 10,000 ppm (malt m kt) Llvtr: adtnomaa oa caaclnomaa (comblntd) E 256 1 --rp------ RM attain: F344 (Fbchtr-344). Moutt Haiku B6CJF,. *Ob*ttvtd offocl comliiro 4 tqulvocali It tody M H Inadequate. Foa butyl btnayl phlhaiata (only), Ih t actual language employed In technical icporl loj (m u Ii rU t w m Npro4iM r u rd n o |M k .N -, i ! ? Ot Gp pp IM S l > S 1 >1 M d si It sr | 1 1I 5i1 {1 2s5| ss i{ i a If Ilf Ii 3 IsO I Uipl 1 I / m s undo 'l " l " IPHIS 41N t r i f W II 18S U H O O 5 I *3 g 4 ? is | a 3 it 1 s I 1. i 3<O1 5| ni fi 1 1 j ill I *s s- lU ill i;i 111 Is sl *f ?* Ila -Il TABLE1 NTTStudia imarprstad asShowingNoCvclnofantc Exacts Oomicai (CASno.) Animal nrain* Roua Dqms TR no. Apr (9002-1KJ) F344 Faad B4C3F, AraMc*um (900041-5) F344 Faad B4C3F, Amoria asbamas (12172-7) Syrian Faad Sridan Amoria sabsmai (12m-73-J) F344 Faad Asriastoa,chrymdla; riwrtand imtrmadlata ran*(12001-29-5) Syrian Faad oidan j Arianna, dimorila; F344 Faad rima ran* (12001-29-5) ! Ariano,crocidoilt (12001-21-4) F344 Faad L-AscorMcasM(SMI-7) F344 Faad 4C3F, mari (119-53-9) 1 F344 Faad BK3F, | ariano! A (8^05-7) F344 Faad HOF, CiyoUcum (10540-2) F344 Faad t 6C3F, iOieraatrianei (107-07-3) F344 Damai Swim CO-1 CJ.addana* 10 (1934-13-0 F344 Faad MC3F, CLacMnri 14 (3SC7-C9-9) F344 Faad B6C3F1 14-Okhlarariantam (93-30.1) brianylaad dadacyi teoria (9002-92-0) POandCyrilaw no. 6 (2713*40) Cwanyl -- (105-07-3) * GOaonria(12002-430) F344 B<OF1 F344 MC3F, F344 uart F344 ic a F , F344 Cava* Faad Faad Gara* Faad 23,000 or 30,000 ppm 25,000or30,000pom IS(Ufarima) 1* (Llfarima) 1 (Ufatima) 230 227 249 279 24< 1%(Ufatima) 293 1%(Ufatima) 2B0 23,000or 50,000 p*m 247 123or 230 ppm(malarats) 230or 500ppm(fornatams) 2300or5000ppm(mica) 1000ar2000ppm(ras) 1000ar3000ppm(matamie) 3000ar 10,000ppm^famaiamiea) 3750ar 7500ppm(ran) 7300ar 15,000ppm(mica) 50ar 100nq/k* (ms) 74 ar 15ma/aidaui (mica) 204 215 214 275 1000ar 3000ppm(rats) 3000ar <000 ppm (mica) <000ar 12400ppm(malams) 12400or23400 ppm(famaiams) 3000ar <000 ppm(mica) <0ar 120m*/h* 2V1 220 2SS 3000ar <000 ppmUrats) <000ar 12400PPm(mica) 12400ar23400 ppm 2S4 20S 1000ar 2000mc/k* (ms) 500ar 1000m|/k* (mica) 20400ar40400 ppm 232 270 2 .' ---^ ;" *- ' 7 'V'SFSf2ii!=g=7: !.. . - __V_-~~-V >---V __ -- 0013405 --. 't- -li.- - - * .. , . - J ) . iycfYS^ BONI 198471 TABLE 1 NT? Studies Interpreted u Showinj No Cardnopenic Effect* (C antm utd) Chemical (CAS no.) Guar cum (9000-300) Hamamelis water (6(916-39-2) HC blue no. 2 (33229-34-4) "nV--M- --W*-ii-w---w-QI-U--I-J--U-T-^ D ta ta , 1.24,74,9and 1 0 4 4 ,7 4 * (57653-45-7 and 19401-74-3) 6 Hydrojcyquinoiine (146-24-3) Locust bean pum (9 0 0 0 4 0 2 ) 0 -Mannitol (69-65-6) Phenol (106-95-2) Propylene (115-07-1 ) Propyl (a lita (121-79-9) Setabim sulfide (7446-306) Setan (trade name, no CAS no.) Stannous chloride (7772-99-6) Tara (urn (39300664) 2,6-Toulenedlamine dlhydrochloride (15461-706) Tremoltte (4567-734) * V--B--f-rU-aB--M- ^B1n^B4n4BV (1,1 (IcMaranhyiraa) (75-354) Animal - strain0 Route P344 B6C3F, F344 B6C3F, F344 B6C3F, Feed Dermal Feed Sw ta Dermal Webster F344 Feed B6C3F, F344 Feed 66C3F, F344 Feed B6C3F, F344 B6C3F, F344 B6C3F, Drinking vsnr Inhalation F344 Feed B6C3F, to t Denta Serba ICK Denta Serba F344 Feed B60F, . F344 Feed B6C3F, F344 Feed B60F, F344 F344 B6C3F, Feed G rata Doses 25,000 or 50,000 ppm ' 50% or 100% In deionized water 5 d/wk 5000 or 10,000 ppm (male rea, male mice) 10400 or 20,000 ppm (female rets, female mice) 0.0 114 .3 times/wk TR no. 229* 266 293 202 1500 or 3000 ppm 25400 or 50,000 ppm 276 .. 221 25400 or 50,000 ppm 236 2500 or 5000 ppm 203 5000 or 10,000 ppm 272 6000 or 12400 ppm 240 0 4 or 14 mi/appHradon, 3 ttmee/wk 043 ml of 2S% or 50% 3 tbnee/ark 197 199 1000 or 2000 ppm 231 2S400or50400ppm 224 250 or 500 ppm (ran) SOor 100 ppm (mice) 200 1(UfetfaM ) 1 or 5 m i/ks ( m ) 2 or 10 mc/kf (mice) 277 22S 'lb s anta: F344 | W w i l l | . Mourn arata: B6C3F,; Sede Webster. ICR Serb* Seta CD-I. HiMHrnfcR SrrtMidUml S3 0 0 1 3 ( 4 0 6 4853 (PWW/JU0 3 ) |I H I i ii 1 3 A JMTi I 9 sa a s e fl?? I 11 aaaaaa ! ? I I ? fi g SSB TA ILS 4. N T f Studili g a fitM at Im 4 |wM ( Orataleal (CAS m .) Antoni train* Routa/doai Riaton for toartoquacy TR MO. A A M M (cktytolUi) and 1,2-AtoiiIk y Ihydraitoi (DMH) (12001-2-$) 1,3,7,t-TCOD and 4 m ilkylbioiaAtkracini (DMBA) I IM M M I Syrian Roldan SwtM-WakUM Iftln d to l(a tfc ifto i) 4 ma/kt By savagi m ry n d n rw u t fer S Anta (DMH) Dannai: OMBA, SOM 1 wfc p ito in TCOO| 0.001 m TCDO Im ta ), OjOOS M TCOO |(MRilct}| p u ippiiciiioHj i ipp U d iim ^ wk Combination uudy to mato and tomato Syrian polka kam tart tw n M rn l inadaipuati kacauM no breralia to OMH-Inducid tolaitlnal naoplatla waaobrarvad (DMH b known in Induci im rotoim tnal tumori) 246 ' Initialion-promolton tiudy using Swbt-WabUar mici 20| waa eomldand Inadaquata bicauta DMBA wat not Halid aloni and DMBA-TCDD-Indirrid (Ibrotarcornai totktonci not graitar ikan Uial okiarvad with TCDD IIqHI *H m tirila: Syrian riMia. Monti attain: Swbt-Wakttir. GO U1 U T tfilZ M O f 0013407 & -H I50 90112158477 3 J.HHASEMANCTau NT? CARONOGENICTT DISCUSSION O f the 86 NT? studies, 50% were found to produce carcinogenic re sponses. Using the results o f the NCI studies reported in TR 1-196 as summarized by Griesemer and Cueto (1980), Hottendorf and Pachter (1982) reported a similar percentage (51%, 98/192 ). They also noted that 28 (15%) o f these studies were found to be equivocal and 66 (34%) demon strated no evidence o f carcinogenicity. When interpreting the results o f these latter studies, Hottendorf and Pachter (1982) note that only three o f the negative experiments were cate gorized by Griesemer and Cueto (1980) as showing no evidence in two animal species. Thus, Hottendorf and Pachter (1982) propose that "the positive to negative ratio in the evaluable bioassays was 33:1." We disagree and feel that this misrepresentation o f the NCI carcinogenesis testing re sults should be corrected. Griesemer and Cueto (1980) actually listed 66 studies showing no evi dence o f carcinogenicity under the conditions used fo r testing and as re ported. They regarded 53 studies as showing no evidence in limited animal experiments, 10 as showing no evidence in 1 animal species, and 3 as show ing no evidence in 2 animal species. When reporting these results, Griesemer and Cueto (1980) stated that "unless the data indicated that the compound had been fully tested in both sexes o f one species at or near the maximum tolerated dose, the evidence was considered to be too limited to draw con clusions about noncarcinogenicity without further tests." Hottendorf and Pachter (1982) apparently misinterpreted this state ment to imply that 63 o f the 66 negative studies were considered by the N Q to be inadequate, or "unevaiuabie." While further testing for certain selected chemicals may have clarified the issue o f whether or not the lack o f a carcinogenic response was due to inadequate dose selection, these studies all employed doses that were estimated to be maximum tolerated doses, often far in excess o f anticipated human exposure levels. Thus, while these negative studies did not " prove" with absolute certainty that these chemicals have no carcinogenic potential, all 66 studies should be considered as being in the category o f no evidence o f carcinogenicity. This was the position o f the N C I, who did not regard these studies as " lim ited." Furthermore, regardless o f the exact " limitations" that may have been present, the studies were n o t " unevaiuabie" as stated by Hot tendorf and Pachter (1982). Each o f these 66 NCI technical reports con cluded that " under the conditions o f this bioassay (chemical name) was not carcinogenic." Studies regarded as inadequate or unevaiuabie by the NCI were dearly identified as inadequate and often were not published as technical reports. Thus, fo r comparative purposes, the actual ratio o f posi tive to negative or equivocal results is approximately 1:1, not 33:1, for both the NCI and NT? carcinogenicity studies. The finding that 19% o f NT? positive studies were based only on in- creased incidences c j from the early NCI The liver was also t often than any oth ! the positive NTP stt corresponding figure NCI studies. The finding tha | surprising, because < metabolized in the I - recognize that mon a carcinogenic resp< mental conditions i i taneous incidence c | corresponding cont , 344 rats (4%), and ) et al., 1984). For } Maronpot and Bot Some authors ( cinogenicity testing rats are sufficient suggest that if one a better choice wc choice would have use o f male and fe ( gens (3 chemicals * A review o f t male rats in conju o f the 98 carcinog 30 carcinogens. D' pose any reductic protocol fo r routir The implicatic | administration ter ) drinking water or 1 contributed to tf carcinogenic respc hydrocarbons tes: chemicals were 2 cinogenic respons were tested utilizi o f studies showin chemical was adm o f administration even if halogenai i s m s lines luce carcinogenic re ad in T R 1-196 as : xndorf and Pachter : They also noted that . uid 66 (34%) demon* lies, Hottendorf and perimems were cate* no evidence in two ) propose that "the s 33 :1 ." We disagree nogencsis testing re* idles showing no evi* or testing and as re* ; ice in limited animal ' ecies, and 3 as show* ae results, Griesemer i th at the compound r near the maximum limited to draw con* 9$ terpreted this state rs considered-by the t testing for certain ;ther or not the lack lose selection, these maximum tolerated posure levels. Thus, solute certainty that . *6 studies should be : o f carcinogenicity, ard these studies as nitations" that may " as stated by H o t -clinical reports conchemical name) was unevaiuabie by the ere not published as actual ratio o f posi* - 1, not 33 :1 , fo r both e based only on in* I creased incidences o f mouse liver neoplasms agrees with the 19% figure * from the early NCI studies as reported by Hottendorf and Pachter (1982). The liver was also the site o f carcinogenicity for the Fischer-344 rat more often than any other organ (Table 5). The finding that 63% (27/43) of the positive NTP studies include liver tumor effects agrees closely with the corresponding figure (64%; 54/85) reported by Hamm (1983) for the earlier NCi studies. The finding that the liver is the primary tumor site in rodents is not surprising, because chemicals administered by feed or gavage are generally metabolized in the liver prior to transport to other organs. One should also i recognize that more than two-thirds o f the chemicals tested did not cause a carcinogenic response in the rodent liver (Table 5), even though experi mental conditions utilizing high doses were employed. Although the spon taneous incidence of liver tumors in male B6G F 1 mice is high (31% ), the ; corresponding control rates in fem ale'B6C3Ft mice (8%), male Fischer- j 344 rats (4%), and female Fischer-344 rats (3%) are relatively low (Haseman et aL, 1984). For a more detailed discussion of rodent liver tumors, see Maronpot and Boorman (1982) and the Nutrition Foundation (1983). Some authors (Von Wittenau and Estes, 1983) have suggested that car cinogenicity testing in the mouse is "redundant" and that male and female I rats a rt sufficient fo r the detection o f most carcinogens. The present data sugpst that if one wishes to halve the number o f sex-species groups tested, | a better choice would be male rats in conjunction with female mice. This { choice would have detected all 43 carcinogens in the NTP data, while the use o f male and female rats would have missed 10 and possibly 13 carcino gens (3 chemicals were positive in mice, but not tested in rats). A review o f the earlier NCI studies produced similar results. Use o f male rats in conjunction with female mice would have detected all but 16 o f the 98 carcinogens; use o f male and female rats only would have missed 30 carcinogens. Despite these results, we feel that it is premature to pro pose any reduction from the current two-sex, two-species experimental protocol for routine carcinogenicity testing. The* implications o f the finding that inhalation and gavage routes o f administration tend to produce more carcinogenic responses than feed, drinking water or dermal exposures are unclear. One factor that may have contributed to the relatively high proportion of gavage studies showing carcinogenic responses is the disproportionately large number o f haiogenated hydrocarbons tested by this route of administration. Among the 86 NTP chemicals were 20 haiogenated hydrocarbons; 80% (16/20) produced car cinogenic responses, primarily in the liver. The majority o f these chemicals were tested utilizing the gavage route o f administration, and the proportion o f studies showing carcinogenic effects was similar, regardless o f how the chemical was administered: 85% (11 /1 3 ) positive studies fo r the gavage route * o f administration, compared with 71% (5/7) for all other routes. However, even if haiogenated hydrocarbons are excluded, the proportion o f gavage 32 0013409 4856 iweg- M * z r M * ? *a t |. IC. HASEMAN ET AC (6 NTT CARCINOGENIC studies showing carcinogenic responses (9/11, 82%) remains elevated rela tive to the corresponding rate in feeding studies (14/43, 3 3 %). in the early NCI studies, 46% (70/152) of the feeding studies resulted in carcinogenic responses compared with 59% (17/29) positive gavage studies. The only other route o f administration with more than one study was intraperitoneal (ip), in which ail 9 studies produced carcinogenic re sponses (Griesemer and Cueto, 1980). Haseman (1984) noted that overall dosed-group survival in the NT? feeding studies has been essentially the same (or slightly exceeded) the survival in corresponding control groups. In contrast, in NTP gavage studies there is often increased m o rality in dosed groups relative to controls. In only 38% (9/24) of the NTP gavage studies were no significant reductions in survival observed in some dosed group relative to controls. One factor that may have contributed to the poor survival in gavage studies is the inherent toxicity o f the class o f chemicals that requires utiliza tion o f this particular route of administration. Generally, NTP does not use the gavage route of administration unless the test chemical is volatile, un palatable, or unstable, or the route mimics a potential human exposure (e.g., drugs taken in single, daily doses). As aiready noted, more than half o f the NTP pvage studies involved haiogenated hydrocarbons, which tend to be hepatotoxic. For a variety o f reasons (including poor survival o f dosed groups), the NTP is limiting the number o f new studies in which pvage is used as a route o f administration. Other disadvantages o f pvage testing include the possibility o f accidental overdoses, the likelihood o f gzvage-reiated trauma (e.g., puncturing the animal's lung, esophagus, or stomach with the gavage needle, thereby debiiiating or killing the animal), possible vehicle (e.g., com oil) effects, and the fact that a single, daily pvage administration (bolus dose)'may not mimic human exposure (Haseman, 1984). A number o f chemicals produced reduced survival in dosed groups showing increased tumor incidence. These included benzene, 1,3-butadiene, chlorobenzene, cytembena (male rats), DBCP, 1,2,-dibromoethane, dimethyl hydrogen phosphite (male rats), DGRE, HC blue no. 1 (female mice) mela mine, 4,4'-oxydianHine (female rats), pentachloroethane, propylene oxide (mice), 1,1,1,2-tetrachloroethane, toluene diisocyanate, trichloroethylene (male mice), and 2,6-xylidine (male rats). However, reduced survival per se does not necessarily mean that the doses employed were too high, since in many cases carcinogenic responses were the direct cause o f reduced sur vival. Moreover, the majority o f carcinogenic effects were n e t associated with a corresponding decrease in survival. Although approximately 50% (141/278) o f N C I/N TP studies have shown carcinogenic effects in laboratory animals, caution must be exercised in the interprention o f this figure. Many chemicals selected fo r testing have a high a priori suspicion o f carcinogenicity. Moreover, since chemicals are not selected randomly, but rather on the basis o f the criteria outlined above, cerain cher figures. Furthermc i twice by different ferent strains of ra theiess suggest tha has been relatively REFERENCES I Ow, K. C- Cueto, C. w mw cardnogeaMoa Grtaaamar, R. A, Mid C ridno for tfw c o f Camrioptn So* Lvon: IARCScion) i Hamm, T. E., Jr. 19S3. , o f U olofleol if*, i York: Plenum. Hot, L. i , Huff, J. E.. March and Mating 2, ad. J.Saxana, pp. Hacmoi, J. K. 19S4. C Hwonai, J. C . Huff, j. intdiw in rodonx. 3 I iitw niiof, G. KL, and tha NationalCancar j Huff, J. E. 1912. Carcb ffaarth H n p o a . *3 ! Hoff, J. E, and Moore. tiouMvahotion at I Made Elom mtn, Um. Huff. J. E* Haaaman, Program, toxicotor tWuationoTDnm Maronpot R. R^ and attarationa and hr ! 0. I Notional Toxicology Pr Human Sorricaa, Pi Nutrition Foundation. A report of tha lir bar 1913. Watidnr Sonag. I. M-, Page, N. OHHS Publication : Von Wlttanau, M. S* i I Fm dam. AppL To; 0013410 y 1* K. HASEMAN ET Al i M NTT CASONOGCNtCmrSTUDIES o MHSIZNOf remains elevated rela1, 33%). ding studies resulted 7/29) positive gavage more than one study luccd carcinogenic re* survival in the NTP lightly exceeded) the in NTP gavage studies Hative to controls* In significant reductions itrols. yo r survival .in gavage Is that requires utiliz*Jly, NTP does not use temical is volatile, un* ittai human exposure toted, more than half Dearborn, which tend o f dosed groups), the h gavage is used as a je testing includeJttie gavage-related trauma mach with the gavage sossible vehicle (e.g., javage administration i, 1984). ival in dosed groups naene, 1 b u ta d ie n e , Dmoethane, dimethyl 1 (female mice) mete* ane, propylene oxide .re, trichloroethylene :duced survival per se ' ere too high, since in :ause o f reduced sur* were nor associated n /N T P studies have ion must be exercised selected for testing over, since chemicals the criteria outlined I ! j 1Ii above, certain chemical classes may be over* (or under*) represented in these figures. Furthermore, these tabulations include certain chemicals tested twice by different routes o f administration, in different species, or in dif ferent strains o f rats. With recognition o f these limitations, these data never theless suggest that the proportion o f chemicals found to be carcinogenic has been relatively constant over the past 8 - 1 0 yr. REFERENCES On , K. C , Gimo, C , a rt Wart, J. M. ISSI, faeton la dw trtuttion of 300 National Cansar Und one ctreinoptn Moaataya. /. ToxkaL Environ. HooMl 1:231-210. Griam a r. K. A^ and Citoto, C 19M . Toward a riirtfln rtn a n em m for di eran of acparfcamnti atidtnet for tha eafclnofenid tv of cftamicait for animan. In Molocuior a rt Ca/Jirtr Axoota of Carcrtopan Scnamhp Tota. eda. R. Moaionnn, H. Banach, and L. Tomatti, pp. 259-2S1. Lyon: iAAC Scientific PuhHririnra. Hamm, T. Jr. 1983. Tha ocairronco of nappiaims In Ion ian k rivo d in . In AopUcottao o f Etacpfcti HmPm to Corsfnofan Totani, oto. H. A. Miman arti S. Sad, pp. 9-23. Now York: n an o. Hwt, L G * Huff, 1 .1 , M oon, ]. A^ a rt ItaU, 0 . f . 1 9 0 . Tha Natiaati TnwirtOfy P rovarti ro* W M wBn| ICBWDVb IIIfMMV AXBBWi OrUOMGO^ yffTOIfVHOBPORIQ^ IQb 2, ed. J. S an a *, pp. 191-244. New Verte A trtele. Haaanan, J. K. 19(4. Pom Itetion tama in artinaeanlaity (aorte. finarte. Appi. ToxieoL, In Hannan, J. K^Hoff, ),, and Boorman, G. A. 19S4. Urn of htatarteti contrai dam in eardnopanieirr atodim in radons. Toxicot. A r t * 12:120*139. Hoeaandarf, G. art Patinar, L J. 19(2. An analyrt of trt c v d m p m tit aorte ncptrtnra of tea National Canear In stata. ToxfeoL A r t * 10:22-2. ta ff, J. E. 1912. Cardnoeonart blnimoy raaultt tarn tea Natiteli Taxlealoey Proeram. Environ. Hoot A rp e a* 45:1(5-19. Huff, J. L , a rt Moon. j. A. 19S4. Cardaopeoart an d ta d a ti* a rt anaartaanrt data tasrpntw dma/mdiiarlon at tin National Teudeoker Pragraaa. In InrtirnW H o u r o f H ootta ned S u r dtetie Oeitomera art. J. Jardttit, P. PMIB, and H. Vatoio, pp. 43 K Now York: Alan X. . Un Huff, U L . Hwaman, J. 1C. McCaimaH, fi. fi* and Moon. J. A. 1MS. Tha National Tajdcalapy Prava, tartcortiy d aa rrtuation rtndaoaa, and tone wem cardnoewwma aaodim. In S oft BmhtoOon o fPrapr a r t atom ico ad. W. K. Uoyd, W talnyan, D .C , Handnato. Haranpoa ( . IL , a rt Boorman. G. A. 19(2. mtorprau tion of radane KraaMeatalar pratifarttiva atantiana a r t hopaeocatfuiar turnon in rtiamlrti ofaty nonane. Toxieoi. A r t * 10:71BQi N attart Toticolofy Ptopram. 19S4. piatti yew 19B4 amwti plan. U . Oapartmene of Haata and Homan t ant e * Putite Hatitii Sarrfea, Withinnon, DjC. Hoattion Poondatian. 19B3. Tha retomen of mmrt Ihw haaamwt hommi arelnom le rta. A report of dM marnational Expan Adrtory Cowmltiao ttm Nuarition Foortation, S raam Bar 19(3. Watitinpton, D .C ; Tha Nutrition Paurtation, Sontas, i. M * Papa, N. P* a rt Saflocti, U. 197. Guidato far raffinav o M alt i M until redan s DHHS PuMtation (N IH ) 7401, National Cane In a ta , Balkda, Md. Von witaanau. M. S * and finta, P. G 19(3. Tha radundwacy of orno raralnopnleliy tioaanye. Tomtom. A ppi. Toxicol. 3 :0 1-4 39. PocohmOHoy 6. 1944 AccopaO / a 26, 196* 0 0 1 3 4 1 1 4858 p .c /'fc T ' 4859 ClinicalT^xicologyof Commercial Products Fifth Edition ROBERT E. GOSSELIN, M.D., Ph.D. Ire n e H e in z G iven P ro fe sso r o f P h a rm a co lo g y, D a rtm o u th M e d ic a l S ch o o l, H a n o ve r, N ew H a m p sh ire ROGER P. SMITH, Ph.D. P ro fe sso r a n d C h a irm a n , D e p a rtm e n t o f P h a rm a co lo g y a n d T o xico lo g y, D a rtm o u th M e d ic a l S chool, H a n o ve r, N e w H a m p sh ire HAROLD C. HODGE, Ph.D., D.Sc. P ro fe ssor a n d C h a irm a n E m e ritu s o f P h a rm a co lo g y a n d T o xico lo g y, S ch o o l o fM e d ic in e a n d D e n tis try , T he U n iv e rs ity o f R ochester, R ochester, N ew Y o rk ; P ro fe sso r in R esidence o f P h a rm a co lo g y a n d O ra l B io lo g y , U n iv e rs ity o f C a lifo rn ia , S a n F ra n c is c o ; o f E n v iro n m e n ta l T o xico lo g y, a n d o fP h a rm a co lo g y a n d M e d ic a l T h e ra p e u tics, U n iv e rs ity o f C a lifo rn ia , Irv in e , C a lifo rn ia With the assistance of JEANNETTE E. BRADDOCK A s s is ta n t in P h a rm a co lo g y, D e p a rtm e n t o f P h a rm a co lo g y, S ch o o l o f M e d ic in e a n d D e n tis try , T he U n iv e rs ity o f R ochester, R ochester, N e w Y o rk WILLIAMS & WILKINS Baltlmore/London 4860 E d ito r: Toni M . Tracy C opy E d ito r: William G. Vinck D e s ig n : JoAnne Janowiak Illu s tra tio n P la n n in g : Wayne Hubbel P ro d u c tio n : Raymond E. Reter NOTE: The first and second editions were authored by Gleason, Gosselin, and Hodge; the third by Gleason, Gosselin, Hodge, and Smith, and the fourth by Gosselin, Hodge, Smith, and Gleason. Copyright , 1984 W illiam & Wilkins A ll rights reserved. This book is protected by copyright. No part of this book may be reproduced in any form or by any means, including photocopying, or utilized by any information storage and retrieval system without permission from the copyright owner. Accurate indications, adverse reactions, and dosage schedules for drugs are provided in this book, but it is possible that they may change. The reader is urged to review the package information data of the manufacturers of the medications mentioned and the ingredients listed on the labels of consumer products. M a d e in th e U n ite d S ta te s o f A m e ric a First edition, 1957 Reprinted, 1958,1960,1962 Second edition, 1963 Third edition, 1969 Reprinted, 1970,1971,1972,1974 Fourth edition, 1976 Reprinted, 1977, 1979,1981 Library of Congress Cataloging in Publication Data Main entry under title: CJiniral toxicology of commercial products. 1. Toxicology. 2. Poisoning. 3. Commercial products--Toxicology. I. Gosselin, Robert E. n . Smith, Roger P. (Roger Powell), 1932- . H I. Hodge, Harold Carpenter, 1904- . IV . Braddock, Jeanette E. [D NLM : 1. Poisoning 2. Poisons. QV 600 G679c] RA1211.C586 1984 615.9 83-1373 ISBN 0^83-03632-7 Preface C oncerning Poisons: Alcohol, hashis tim e.--R a lp h 1 Cocaine isn't h h ilah B ankh ec C oncerning Consume The consumer There is no saf C oncerning Science: There is somet conjecture out Hindsight is th C oncerning T ru th : ! As scarce as tn W heeler S h a n C oncerning Books: ' ; In this work, v. that much like Books are help The original purpos effectively w ith acute The book provides (a) addresses and telephor not available, (c) sam toxicity of each formu o f individual ingredien W e suggest th a t the in the seven sections c to U se T h is M a n u a l hypothetical cases in described below. Over the years a s emphasis, namely to a nisms induced by van* detailed documentatic research papers as we sional toxicologist. Sc some extent in Sectio: this edition does not uncertainty or disagrgaps in knowledge ant - -.... pmphasis is on acute t V- W 7 48R1mutagenic effects hav ' SECTION V. TRADE NAME INDEX V-235 SALTS SPECIAL ver s(Bifluorides) '-5 ^ :OCIN STEARATE - A B >. l-.l n stearate 125 mg.; 250mg; 500mg. lOMAST 36 MAMMARY ION veterinary ribusiness) -` vi . e: _ ein.......... 300mg. STRIPPER IC" PROFESSIONAL C SCAPE STRIPPER AUTIFYING TINT -' rs', embalmers' supplies r -s') T- tydrocarbons ' , :Y SHAMPOO rs', embalmers' supplies -s') ydrocarbons * UID COLORING rs', embalmers supplies s ') \KPROOF SKIN s's)', embalmers' supplies ydrocarbons * ;UID POWDER V, embalmers'supplis E SC O T Antacid (DirectDiv.) ESTEAM - E.S. (Powder) SeeG U N K ES. -ESTEAM (Powder) Each capsule: Bismuthalumnate *t 100 mg. Magnesium trisilicate * . . 160mg. CoprecipitateofAluminum hy ESTEE LAUDER COSMETIC PRODUCTS droxideand Magnesium carbonate * ......... 130 mg. Estee Lauder Inc. 767 FifthAve. tSeeBismuth salts New York. N.Y. 10022 ESTERON FOUR WEED KILLER Herbicide MDow) Activeingred.:.......... 72.8% 2,4-Dichlorophenoxyacetic acid, butoxy propylester ' Inertingred.: ............. 27.2% Emulsifiers Petroleumsolvent * Phone: 212-572-4200 E.S. - ESTEAM (Powder) See G U N K E.S.-ESTEAM (Powder) See COSMETICS, Section VI, General Formulations ESTERON 245 HERBICIDE :<Dow) 2,4,5-Tbutyletherestersofpropyl ene glycol (2,4,5-T add equiva- ESIDRIX ESTEEM, KLENZADE lent45.0%) *f ........... 69.2% Inertingred.: ............. 30.8% Diuretic-antihypertensive See KLENZADE ESTEEM Petroleumsolvent ........ 27.3% (CIBA Pharmaceutical) fSeeTrichlorophenoxyaceticadd Each tablet: Hydrochlorothiazide * 25 mg.;50mg. ESTERCOL For shin irritation (Torch) ESTERON 44 IMPROVED ESKALITH CAPSULES Mercuryoleate * ......... Control of manic episodes in manic-de Phenol (Carbolicacid) ...... 0.45% 0.5% W EEDKILLER (Dow) pressive psychosis Salicylicadd * ............ 3.0% 2,4-Dichlorophenoxyaceticadd,bu (SK&F) Coaltar * ............... '1.2* tylesters * ............. 51.0% Each capsule: Lithiumcarbonate * ..... Oilemulsionvehide .... 300 mg. ad. 100.0cc. 76 Ed. Inertingred.: ............. Emulsifiers Petroleum solvent * 49.0% ESKALITH CR TABLETS ESTERON 76 BE HERBICIDE Control of manic episodes in manic-de .(Dow) pressive psychosis (SK&F) 2,4-Dichlorophenoxyaceticacid,bu ESTERON 10-10 WEED KILLER (Dow) Each tablet: Lithiumcarbonate * ..... 450 mg. tyl esters (2,4-dichlorophenoxyaceticacidequivalent-63.2%) * 79.2% Inertingred.: ............. 20.8% Petroleumsolvent ........ 16.8% Activeingred.: ............ 72.8% 2,4-Dichlorophenoxyaceticadd, propyl ene glycolbutyletheresters * Inertingred.: ............ 2721% ESKALITH TABLETS Control of manic episodes in manic-de ESTERON BRUSH KILLER Emulsifier Petroleumsolvent * pressive psychosis : (Dow) (SK&F) Each tablet: Lithium carbonate * ..... 300 mg. 2,4-D propylene glycol butyl ether esters ............... 36.0% 2,4,5-Tpropyleneglycolbutylether esters * ............... 34.1% ESTERON 99 WEED KILLER Herbicide VfDow) ESOTERICA CREAM lar, Facial, Fortified) Fade cream (NordiffThayer) (Regu Inertingred.: ............. 29.9% Petroleumsolvents * ...... 26.4% ESTERON 245 CONCENTRATE 2,4-Dichlorophenoxyaceticadd, propylene glycol butyl ether esters * ............... 41.0% Inerts: .................. 59.0% PrindpallyPetroleum solvent * Hydroquinone * ................................. 2.0% ..Herbicide UDow) ESTERON 99 WEED KILLER ESPOTABS Laxative (Combe Inc.) Yellow Phenolphthalein 2,4,5-Trichlorophenoxy acetic acid, propylene glycol butyl ether esters * ............... 92.5% CONCENTRATE -Herbicide JDow) Inertingred.: ............. Petroleumsolvent ........ 7.5% 2.5% 2,4-Dichlorophenoxyaceticadd, propylene glycol butyl ether esters * ............... 72.8% Inerts: ......... ........ 27.2% esq uire sh o e care PRODUCTS (K nom ark) Knom ark, Inc. 132M errick Blvd. Jam aica, N.Y. 11434 ES(TDoEw)RON 6E HERBICIDE PrindpallyPetroleumsolvent * 2,4-Dichlorophenoxyacetic add, isooctyl ester (2,4-dichlorophenoxyacetic add equivalent 62.6%) * ............... 94.4% ESTES NU-RAL TABLETS Analgesic (Estes) pbone:212-276-3400 Acetylsalicylicadd * Caffeine........ 5gr. 1/6gr. Starred ingredients (*) may be responsible for major toxic effects; consult Section IL 4862 n s 4863 tt-UUUXl- ( J GO) .. 7w J Dioxins have left their mark on ch il dr en from Seveso nine years ago.... THE LINGERING (INSIDIOUS) CONTAMINATION* S c i e n t i s t s ( i n v e s t i g a t o r s ) h a v e now d i s c o v e r e d how d i o x i n s work in the > D0M2159520 The mo t o r is an ideal d i o x i n - f a c t o r y . This fact from U l m ' s dioxin-analyst Professor Karlheinz Ballschmiter has aLarmed the a u t o m o b i l e industry. With his colleagues, Dr. Heinz Buchert has demonstrated: Autos produce highly poisonous dioxins and furans. In the exhaust of 24 German automobiles, the chemists have identified these materials. With soot the dioxins are blown into the air, and they a c c u m u l a t e in the di rt of the s t r e et . One gr a m of dirt (dust) can co ntain up to 120 bi ll io n parts of a gram (nanograms) of the Seveso dioxin 2,3,7,8 TCDD, as we lL as other dangerous dioxins. We wouLd probably all be long dead,* says Dr. Buchert, *if the sun did not to a great extent break down this material.' Sources of the danger are permissible mixtures, that make leaded gas antiknock and motor oil long-lived, as well as mixed PCB's, wh i c h a r e p r o h i b i t e d in r e c y c l e d moto r oil and w e r e used for i n s u l a t i o n in t r a n s f o r m e r s un ti l 1983. The e x p e r t s of h e a L t h and e n v i r o n m e n t a l a g e n c i e s in m a n y industrial countries have recognized the general dioxin danger. As^with DDT, the Seveso-poison has now also spread over the n o r t h e r n h e m i s p h e r e . B e a r s at the A r c t i c Circle, h e r r i n g s in the East Sea, crabs from Canadian waters, salmon and carp from Swedish ri ve rs and lakes, mo re ov er pork fat, c h ic ke n liver, or cow's milk are already tainted with this and other dioxins. In G e r m a n y d i o x i n s and f u r a n s c o n c e n t r a t e in the s e d i m e n t s of the Rhine, Neckar and Bodensee, as the analytical Professor HanspauL Hagenmaier from Tubingen has discovered. On the B o d e n s e e th is c o n t a m in a t i o n b e g a n im m e d i a t e l y af t e r WWII wi t h n o n b io d e g r a d a b Le m o L e e u les fr o m r e t o r t s (coke o v e n s ) . At that ti me chemicaL pLants first manufactured the defoLiant 2,4,5-T, as well as DDT, Lindane and PCP, thereby co-producing dioxin. The dangerous materials are by no means, as industry asserts, the companions of mankind since the 'origin of f i r e . 1 Th e m a i n p r o d u c e r s of the w o r l d w i d e c o n t a m i n t ion, in the VS* ||| r ^ o p i n i o n o f s c i e n t i s t s and e n v i r o n m e n t a l groups, is the ot he r s e g m e n t of *the c h e m i c a L i n d u s t r y w h i c h p r o d u c e s w o o d and pl an t prservtives containing these materials. If dioxins arise from the smokestacks of incineration estab lishments, they are predominantly traced back to combustion of waste containing problem chemicals such as PCP, report v? Canadian authorities. *The key to the environmental problems,* says Professor B a l I s c h m i t e r , 'are dioxins and furans. If we <3c o u l d s u c c e s s f u l l y r e d u c e t h e s e m a t e r i a l s , m a n y p r o b l e m s w o u l d be *olved-' 0010857 W h i L e e n v ir o n m e n t a L ists *+ i L t for the most part u n s u c c e s s f u l ly b a t t l e s i n g l e c h e m i c a l produc ts , such as lindane, DDT, or PCP, industry w o r l d w i d e has long re cognized that, above all, the true <s u b s t a n t ia L ) d a n g e r t h r e a t e n s via the d i o x i n in v e s t ig a t i o n s of the h e a l t h and e n v i r o n m e n t a l p r o t e c t i o n agencies of Canada, the U.S., Sweden, the Netherlands, and Italy. Ther ef or e, the A m e r i c a n c h e m i c a l giant Dow C h e m ic al took up the p u r s u i t itself. Its a n a l y t i c a l te am L a m p a r s k i/T.jnestr ick (T.J . N e st ri ck) has just disclosed at the 5th International D i o x i n S y m p o s i u m in B a y r e u t h , that the co mpany, th ro ug h the production of the defoliant 2,4,5-T, has contaminated the soil of the city of Midland, Michigan, with up to 450 m i c r o g r a m s per kilo <ppt> of S e v e s o - p o i s o n . On the co mpany property, up to 52,000 ppt have penetrated the ground. By comparison: on the p r o p e r t y of the p r e v i o u s l y c l o s e d c h e m i c a l plant B o e h r i n g e r in Hamburg one finds more than one milLion ppt. No German company has yet admitted, as openly (frankLy) as the Dow C'hemicaL Co., what risk its p r o d u c t i o n enta iIs for the e n v ir o n m e n t . The Seveso-TCDD can escape (out-gas) from the ground and spread through the air and a Iso into the water. The Canadian EPA therefore warns against eating more than one fish per month out of Lake Ontario. In the body fat of Canadians, as well as Americans, up to 1000 ppt d i o x i n s and furans have aLre ad y accumulated. Included therein are up to 10 ppt Seveso-poison . On the ot h e r hand, in G e r m a n y the federal he a L t h b u r e a u knows virtually nothing about the burden to the population. S u i t a b l e i n v e s t i g a t i o n s s h o u l d be s t a r t i n g in a few mo nt hs . H o w do t h e s e p o i s o n s wo rk in the b o d y ? H o w mu ch can a pe rs on take up daily? In that vein, the ideas (opinions) of industrial toxicologists and scientists of the U.S. EPA were b o u n c e d a r o u n d ( d i s c u s s e d ) in B a y r e u t h . The EPA LabeLs the Seveso-dioxin as an initiator and at the same time as a promotor of cancer. Certainly, for this reason, the d o s e s h o u l d b e o n l y zero. T h e r e f o r e , it s e e m s that s o c i e t y accepts a certain number of cancer victims through this ma ter ia L. t Industry and the federaL heaLth bureau favor the theory that this poison at best potentiates the cancer-producing acitivity of other chemicals such as arsenic. Thus one couLd without danger daiLy ingest and inhale dioxin. Uhat a person accumulates today, he can only after decades w h o l L y e l i m i n a t e (excrete), d i s c o v e r e d the Z u r c h e r (?) toxicologist Dr. Hans Poiger through an experiment upon himself. Canadian and American biochemists have discovered that the Z96SI ma te r i a l is d a n g e r o u s in s m a l l do se s. Hamste rs g u i n e a pigs, rabb it s, sheep, R h e s u s m o n k e y s , wh ic h are e s p e c i a l l y se ns it ive to dioxin, as well as at least 1 0 - 2 0 % of people, h a v e in c e r t a i n cells a protein, to which the Seveso-poison and similar substances bind. Indeed this so-caLLed A-receptor induces the production of enzymes, which should detoxify the body. However, the enzymes cannot break down the unnatural molecules. The excessive enzyme production now serves as a 'surveyor's rod* (marker) for the inherent danger of dioxin-containing chemicals. Under the influence of dioxin, the A-receptor even invades the genetic material. It al te rs the genes of other growth factors, of maturation as well as differentiation of replacement c e l l s in the skin, and of the r e g u l a t i o n of the t h y m u s gland, which belongs to the immune system. The results : chloracne, as well a s fewer active T-lymphocytes, the police of the body. That in r e t u r n m e a n s w e a k e n i n g of the immune sy st em . T h e o r g a n i s m becomes a victim of bacteria, viruses, as well as cancer. Moreover, dioxins cause a vitamin-A deficiency which can eventually Lead to cancer. Under the action of dioxins, the thyroid gland produces fewer hormones-...T h e r e b y n o d u l e s (g ro wt hs ) are fo rmed. M e t a b o l i s m is dist ur be d, and d e p r e s s i o n o c c u rs . In c o n c l u s i o n , d i o x i n s a r e the r e a s o n (cause) for t h i s -- as d e t e c t e d in mice, too few h o r m o n e s are al so p r o d u c e d in the o v a r i e s and f e r t i l i t y d inunishes. Nevertheless (meantime), scientists have discovered new moLecuLes, which are similar to S e v e s o - d i o x i n , but which should be stiLl more poisonous. Professor Ballschmiter believes therefore: 'that we only stand at the beginning of a Laborious search for uLtra-poisons. Out of the exhaust and out of the incineration facilities come thousands of things, about which we still know nothing at all. (No end of m o t e . ) written by ELVIRA SPILL, STERN magazine translated r e sp ec tf ul ly by Anit a S. Knight and L.P. McCarty, 11-15-85 f 4866 0010859 Dioxine haben vor neun Jahren Kinder von Seveso gezeichnet. Doch die Substanzen sind, aus vielen 4 4t DQW2IS952J Verseuchung m Wissenschaftler haben jetzt herausgefunden, wie Dioxine im Krper wirken Der Motor ist eine idea| te Dioxin-Fabrik. ' Diese Feststellung des L'lmer DioxinAnalytikers Professor Karl heinz BaDschmiter schreckte die Autoindustrie. Mit seinem Kollegen Dr. Heinz Buchen hatte er nachgrwlesen: Autos produzieren bochgiftige Dioxine und Furane. In den Auspuffaniagen von 24 deutschen Automobilen hatten die Che miker diese Substanzen identi fiziert. M it dem Ru werden die Dioxine in die Luft geblasen und lagen] sich im Staub der Straen an. Ein Gramm Staub kann bis zu 120 bil&onstel Gramm des Seveso-Dioxins 2,3.7.8-TCDD, aberauchande re geflhitiche Dioxine enthal ten. Wir wren wahrschein lich aOe schon lngst tot, sagt D r. Buchen, wenn nicht die Sonne einen groen Teil dieser Stoffe wieder zentne. Ouee der Gefahr sind er laubte Zusatzstoffe, die Blei benzin Tfpffbstund Motorl langlebig machen, aber auch verbotenen in wieder aufgearbeitetes Motorl gemixte polychlorierte Biphenyle (PCBs). die bis 1983 zur Isola tion in Transformatoren ver wendetwurden. Die Experten der Gesundheits- und Utnwehbebrden vieler Industrielnder haben . die generelle Dioxin-Gefahr erkannt. Dean wie einst D D T hat sieh jeta auch das SevesoG ift vor allem Ober die nrdli che Halbkugel verbreitet. B ren am Polarkreis. Heringe in der Ostsee. Krabben aus kana dischen Gewssern, Lachse und Karpfen aus den Flssen und Seen Schweden, aber auch Schweinefett. Hhnerieber oder Kuhnrikb and bereits mit diesem und anderen Dmo- n + n SlacfRt ne in den Sedimenten von Rhein. Neckar und Bodensec an. wie der Analytiker Profes sor Hanspaul Hagenmaier aus Tbingen herausfand. Am Bo densee bat diese Verseuchung mit Moleklen aus der Retor te. die von der Natur nicht ab gebaut werden knnen, unmit telbar nach dem Zweiten Welt krieg begonnen. Damals hat ten zum etstenmal Chemie-Fa briken das Entlaubungsmittel 2.4.5-T hergesteUt. sowie D D T. Undan und PCP - und dabei Dioxin mitproduziert. Die gefhrlichen Stoffe sind also keineswegs, wie die lndu-strie -behauptet, -von Anbe ginn des Feuers Begleiter der Menschen. Hauptverursacher der weltwehen Verseuchung, so die Meinung von Wissen schaftlern und Umwehbehr den. ist jener Teil der chemi schen Industrie, der Holz- und Pflanzenschutzmittel mit die sen Stoffen henteOt. Wenn Dioxine aus den Schloten der Mllverbren nungsanlagen quellen, sind sie Professor Kflfette BallsdunttarmUla vorwiegend auf die Verbren nung von Abfall mit ProblemChemikalien wie PCP zurck zufhren. urteilen kanadische Behrden. Der Schlssel fr Umweltprobleme. so Profes sor BaDschmher. sind Dioxi ne und Furane. Wenn es uns gelnge, diese Stoffe zurckzu drngen. wrde sich vieles von sefest erledigen. Whrend Umweltschtzer noch immer und meist erfolg los einzelne chemische Pro dukte wie Lindau. D D T oder PCP bekmpfen, hat die Indu strie wehweit lngst erkannt, da ihr wirkliche Gefahr vor allem durch die Dioxia-Untnsuchungen der Gesundheits und Umwehschutzbehrden Kanadas, der USA. Schwe dens. der Niederlande und Italiens droht. Der amerikanische ChemieRiese Dow-Chemical ergriff deshalb die Flucht nach vorn. Seio Analytiker-leam Laropanki/Tjnestrick offenbarte jeta auf dem 5. Internationa len Dioxin-Symposium in Bay reuth. da die Firma durch die Produktion des Entlaubungs mittels 2.4.5-T die tr a e der Stadt Midland im US-BuodesStaat Michigan mh bis zu 450 milliardstel Gramm pro Kik> ippt) Seveso-Gift verunreinigt hat. A uf dem Firmengelande seien bis zu 52 000 ppt ins Erd reich gedrungen. Zum Ver gleich: Auf dem Gelinde der inzwischen geschlossenen Che miefabrik Boehringer m Ham burg finden sich mehr ah eine Milhon ppt. So offen wie Dow-Chemical hat noch keine deutsche Firma dokumentiert, welches Risiko ihre Produktion fr die Um weltbedeutet.___ Das Scveso-TCDD kann aus der Erde ausgasen und sich durch die Luft, aber auch im W m er awbreiten. Die kana- mesiZHoa ClMCM IUkU U UC1IIUllMIM' Ul L4i'ttMlC4iM S444 See pro Monat zu essen. Im Er vcrinden die Gene jenes Krperfett von Kanadiern. Wachstumsfaktorv der Rei- aber auchvon Amerikanern ha- fung sowie Teilung von Nach* ben sich bereits bis zu 1000 ppt Khuhzrllen fr die Haut und Diorine^und Furane antesam* .der zum Immurtsystcm geb- men. Darunter sind bis zu 1 0 * renden Th>mus-Drse steuert* ppt.Seveso-Gifl. Die Folge: Chlorakne, aber In der Bundesrepublik wei auch weniger aktive T-Lym das Bundesgesundheitumt da* phozyten. der Polizei des Kr fegen so gut wie nichts Oberdie pers. Das wiederum bedeutet Belastung der Bevlkerung. Schwchung des Immunsy- Erst in einigen Monaten soll mit stems. Der Organismus ist Bak entsprechenden Untersuchun terien. Viren, aber auch Krebs gen begonnen werden. hilflos ausgeliefen. Auerdem Wie wirken nun diese Gifte lsen Dioxine einen Vit im Krper? Wieviel darf ein amin-A-Mangel aus. was eben Mensch tglich aufnehmen? falls zu Krebs fhren kann. Darber prallten die Meinun Unter Einwirkung von Dio- gen von Toxikologen der Indu xinen produziert die Schild strie und Wissenschaftlern der drse weniger Honnone. Da amerikanischen Umwchbehr- durch bilden sich in ihr Kno de EPA in Bayreuth aufeinan ten. Der Stoffwechsel ent der. gleist. und es kommt zu De Die EPA stuft das Seveso- pressionen. Dioxine sind Dioxin als Initiator und zu- schlielich die Ursache dafr. gleich ab Verstrker von Krebs da - wie an Musen nachge ein. Sicher sei deshalb nur die wiesen - auch in den Eieistk- Dos nuU. Es sei dem. die Ge ken zu wenig Hormone geb- sellschaft akzeptiere eine ge wisse Zahl von Krebsopfern durch diese Substanz. Die Industrie und das Bun desgesundheitsamt favorisieren die Theorie, da dieses Gift hchstens die krebserregeadc Wirkung anderer Chemikalien wie etwa Arsen verstrke. Abo knne man ohneGefahr tglich weiterhin Dioxin mitessen und einatmen. Was der Mensch beute an Diomnen speichen, kann er ent nach Jahrzehnten vollends wieder ausscheiden. fand der Zrcher Toxikologe D t. Hans Poiger durch einen Selbstver such heraus. Da dir Substanz auch in geringer Dos gefhr lich ist. haben kanadische und amerikanische Biochemiker entdeckt. Hamster. Kanin chen, Hhner. Schafe. Rhesus affen. die gegenberDioxin be sonders empfindlich sind, aber Proftssor Hampaul Hagsiunaier aus Tfibtagea tntdeckte D io xin I Schlamm am Grand das Bodiftsats auch mindestenszehn biszwan zig Prozent der Menschen ha ben in bestimmten Zelten einen det werden und die Fruchtbar Eiweikrper, der das Seveso- keit nachlt. Gift und hnliche Substanzen Inzwischen haben die Wis an sich bindet. senschaftler neue Molekle Dieser sogenannte Ah-Re- entdeckt, die dem Seveso-Di zeptor lst zwar die Produktion oxin sind, aber noch von Enzymen aus. die den Kr giftiger sein soflen. Professor per entgiften sollen. Doch die Balhchmiter glaubt deshalb. Enzyme knnen die naturfrem- da wir eist am Anfang einer dea Molekle nicht knacken. mhscEgen Fahndung nach U l- Die Ibenniife Enzympro tngiften stehen. Aus dem duktion dient nun ab Melatte Auspuff und aus den MDver- fite dfe Gefhrlichkeit von brennungsaniagca kommen dHfxinh tftigf n Tausende von Stoffen. Ober Unter dem Einflu von die wir bislang berhaupt noch Dioxin greift der Ab-Rexepior CLvnu \ rS> C O O lflftfif Z7/ **4 4869 > a i RCIZHOU < 1. h - i It1 -----^7 1 C D d C C r'i- j / -- j ' t ERATOGENESIS c ARCINOGENESIS MUTAGENESIS 4870 VOLUME 5, NUMBER 4,1985 P - ^ to U ALAN R. LiSS, INC. < s tnd M ity of 'sum . Pathol -body duced 1975. nonounder .uman USA' , EL: Ikins. unda- 1. pp dene mine : Biol >and tied . and mide jenic luu( t- \ l XL -- y / C -D O r [ - C L - L - i / - C ' <'C C L/ S J Teratogenesis, Carcinogenesis, and Mutagenesis 5:231-250 (1985) R e p ro d u ctive E ffe c ts o f H e rb icid e E xp o su re in V ie tn a m : R ecent Studies by the V ietnam ese and O thers John D. Constable and M aureen C . Hatch Ambulatory Care Center 3A-3S3, Massachusetts General Hospital (J.D.C.), Boston, Massachusetts, and the Division of Epidemiology, Columbia University School of Public Health, New York, New York (M.C.H.) Key words: Agent Orange, hydaddifbrm mole, birth defects, fetal km . Vietnam IN T R O D U C T IO N Are there adverse effects on the children o f individuals exposed to such herbi cides as Agent Orange and its contaminant 2,3,7,8-tetrachlorodibenzo-p-dioxin? This weighty question continues to be debated here in the United States and elsewhere, in the courts as well as in scientific meetings. Initially, concern about possible harm to exposed human populations was prompted by observations in the laboratory: specifically, teratogenic effects among die offspring o f treated females and some testicular toxicity in treated males. From the experimental data it was also apparent that the variation in response across species (small mammals, nonhuman primates) was considerable. Hence direct evidence from studies in man is clearly essential to answer questions about the reproductive risk to humans. Regrettably, such data are sparse and frequently poor in quality, [see refs. 1-4 for critical reviews o f the literature]. Two relevant studies o f reproductive outcomes among U .S . soldiers serving in Vietnam have recently been published [5 7]. We discuss these latest reports briefly below, but they are of course available to anyone wishing to peruse and evaluate them further. The principal objective o f this paper is to put before the interested reader as full an account as possible o f related but unpublished epidemiologic investigations carried out by Vietnamese researchers. These studies, which are unlikely ever to appear in Western scientific journals, were presented in January 1983 at an International Sym posium on the long-term effects o f phenoxy herbicides, held in Ho Chi Minh City (formerly Saigon) and attended by the authors along with some 70 other visiting scientists from the U .S ., the United Kingdom. Western and Eastern Europe, Asia, and the U .S.S.R . Because o f the continuing debate, it seemed to us important to bring these studies to the attention o f the wider scientific community and to provide as Addren reprint peque to Maureen C. Hatch, Division of Epidemiology. Columbia Univenity, School of Public Health: 600 West 168th S u m . New York. NY 10032. 1985 Alan R. Uss, Inc. 0010813 ItR O IlM R S 2 3 2 Constable and Hatch complete and unprejudiced a description of their methods and findings as possible, such that readers may make some independent assessment o f the evidence contained therein. The studies oudined below were presented formally to the Symposium's Repro ductive Epidemiology Working Group,* o f which one o f us was rapporteur (J.O.C.) and the other a participant (M .C .H .). A ll conferees were assigned to one of eight such groups for in-depth discussion o f the Vietnamese research. We were provided with English translations o f each paper upon our arrival in Vietnam. At subsequent meetings o f the working groups, the authors o f the investigations presented their studies and responded to questions from the other participants. The descriptions we provide draw on information elicited in these discussions as well as on that contained in the papers circulated to us. (See subsequent footnote for fuller details.) Tables o f dam are presen ted exactly a s they appear in the original reports. Wherever we have made additional computations, this fact is duly noted in the text. METHODOLOGICAL PROBLEMS IN VIETNAM ESE STUDIES OF THE REPRODUCTIVE EFFECTS OF HERBICIDE EXPOSURE Before analyzing the Vietnamese investigations individually, it may be well to consider some general problems that apply to many or all o f them. Two types o f studies were carried out in Vietnam to assess the reproductive effects o f herbicide exposure: 1) Studies in the South compared reproductive out comes among couples living in sprayed areas with those among couples living in unsprayed areas. 2) Studies in the unsprayed North compared reproductive outcomes among unexposed women whose husbands served in the South, and hence were potentially exposed, with those among women whose husbands remained in the North. Specific problems which may affect either type o f study include the following. Bias in Selection an d /o r Reporting Bias is a threat to the validity o f any scientific research, particularly so when the investigation does not take place in the pristine environment o f the laboratory but in the real world. Hence it is unfortunate that many o f the Vietnamese papers prevent adequate evaluation because they are less detailed than one would wish in describing the selection o f subjects and the method o f data-gathering. The Vietnamese research ers do, however, appear to be aware o f mthodologie issues. In one investigation (9] the Northern districts chosen for study and the affected children selected as cases were determined using a truly random method. In another study carried out among Northern women in respect to exposure o f their husbands (10] it was appropriate that the team determining the nature and frequency o f reproductive problems was different from that inquiring about potential herbicide exposure. Blind interviews were more difficult to cany out in the South where exposure status is based on residence; knowing the respondent's domicile meant that the interviewers usually knew whether they were talking to an exposed or unexposed subject This caveat in respect to studies ------------- t > A votante based on the proceedings of die Symposium has recently been published (Westing AH. ed: "Herbicides in Wan The Long-term Ecological and Human Consequences." London and Philadelphia: Taylor and Francis. 1984. 210 pp.) The volume contains a chapter on reproductive epidemiology, which includes only a telccted few of the papers presented to the Working Group, along with a brief discussion which draws on (he material in our present, longer review. { 'ssible. uained R epro*.D.C_.) rf e ig ht 'OvidiCL s e q u e flt d thel? ons vtfe nam ed tbles /e havfe well to Juctive fz outr ' in u. .nes : were North, w in g . when ry but revent ribing earchon [9] cases unong te that ferent more Jence; hether xudies H . ed: elptoa: w h ich v--TM Herbicide Exposure in Vietnam 233 carried out in the South may not apply to the investigations at a Ho Chi Minh City hospital, since the patients' residence would not be so readily apparent as in the case o f home interviews. A true double-blind study where the respondents as well as the interviewers are blind to exposure cannot even reasonably be proposed under the conditions prevailing in Vietnam. D efinition of Exposure In virtually all o f the studies to be discussed, designation o f exposure is based on residence in an area sprayed with herbicides as determined by history and/or visible forest or crop destruction. Today the presumption o f exposure could be confirmed by US military flight records which have become available (ie. the Defense Department's HERBS tapes). Obviously there must have been enormous variation in individual exposure; some people were outdoors at the time o f spraying, others indoors, others away from the village for a day or two. Some merely passed through a sprayed area; others continued in residence there. Excepting one study where only preliminary results have been reported [11], no attempts have been made to construct an exposure index, nor has a distinction been drawn between direct exposure or secondary exposure through the diet. In no instance have the various herbicidal agents been differentiated. The studies carried out in the North, while effective in isolating paternal exposure from exposure o f the mother, consider all Northern soldiers who were posted to the South as exposed, although presumably only some o f them actually were, at least to any significant degree. The inclusion o f all w ill weigh against the demonstration o f health effects o f herbicide exposure in the studies in question. Exposure o f the female only cannot be studied, since women living in sprayed areas generally had husbands living with them who.were similarly exposed. No effort has hitherto been made to seek out exposed women with unexposed husbands, and this may not be feasible. Hence, while certain effects have been ascribed to maternal exposure only, paternal influence cannot be excluded. Sources of Data on Reproductive Outcom es Four pregnancy outcomes have been examined in the Vietnamese studies: miscarriage, stillbirth, congenital defects, and hydatidifbrm mole, or molar pregnancy. The frequency o f miscarriage has been ascertained solely by verbal repon rather than through medical records. W hile this method is considered acceptable and is the only one used in many worldwide studies, in one o f the Vietnamese papers [11] the data on miscarriage are somewhat inconsistent, suggesting that these self-reports may not be entirely reliable. Stillbirths were generally ascertained from health records. It is believed that at least 75% o f all births in Vietnam take place within the Ministry of Health system, ie, at least within the knowledge o f a rural midwife. Original midwives' records provide reliable data on number o f live and stillbirths, sex, and weight o f infant and placenta. However, the noting o f congenital anomalies is very erratic, even though there is a column provided for recording them. In these studies reported birth defects were validated in so for as possible. Local health records were checked and surviving children with malformations were examined by physicians from the investigative team. In most cases, specific congenital anomalies appear to have been carefully observed and listed. However, their classification is not uniform. A few o f 4873 MlfflSlS S iH S U M O B t ft- 6 2 9 6 S f 2 MOQ 234 Constable and Hatch the studies (12,13] include among congenital anomalies aberrations of viral origin, such as infantile poliomyelitis, and obscure conditions such as enuresis. It is impossible to be sure how complete the ascertainment of birth defects has been in these investigations, but the way the inquiries are set up makes this unimpor tant, providing the search was uniformly carried out in both groups: exposed and unexposed. The overall rate o f congenital anomalies reported among the unexposed is at the very low end o f the worldwide range (ie, 0.5% ). Prior to the herbicide spraying there had been no real effort to establish the expected rates o f various congenital anomalies in Vietnam, making evaluation o f the figures presented in these studies difficult. To provide some context for their assessment, we include here some data from a W HO survey o f several Southeast Asian nations carried out in the early 1960s (Table I). As the table shows, the overall rate o f common congenital malfor mations in these populations is similar to that reported from Vietnam. TABLE I. WHO Compandre Study of Congenital Maifonnadoos (SuppL 34 Bull. W HO, pp. 1127, Geneva. 1966)________________________________________________________________ Raie of common congenital malformations in the ____________Centre (per 1,000)____________ t 2a 2b 3a 3b 4 Note Down's syndrome Ancncephahis Did A SB Hydrocephalus DtoandSB Spins bifids Occipital SB Other NTD (A flN TD ) Oesophageal atresia Anai atresia Other gut malformations Exomphaios Q eftlip e ft lip + palate e ft palate Talipes Polydactyly U Other poiydaayiy Radial poiydaayiy Syndactyly Other dighai Rcducooo dtionuiw t Other limb deformities Sirenomelia Conjoined twins Total per 1,000 deliveries 0.17 1.24 0.00 0.29 0.10 0.26 0.00 0.07 (1.96) 0.00 0.10 0.20 0.20 0.41 1.01 0.20 1.62 0.00 0.61 0.91 0.10 0.20 0.00 0.10 0.10 0.00 7.89 0.00 1.74 0.13 0.73 0.29 0.75 0.04 0.03 (3.75) 0.08 0.30 0.00 0.00 0.32 0.09 0.03 0.00 0.13 0.00 0.03 (0.62) 0.00 0.10 0.00 0.20 0.35 0.73 0.13 0.96 0.10 0.25 0.023 0.00 0.03 0.10 0.35 0.00 0.00 7.58 0.03 O Jl 0.31 0.16 0.42 0.10 0.26 0.00 0.03 0.00 0.03 0.26 0.00 0.00 2.69 0.16 1.00 0.06 1.04 0.00 0.19 0.00 0.00 (2-29) 0.06 0.56 0.19 0.12 0.31 1.25 0.00 1.63 0.19 0.13 0.25 0.06 0.12 0.13 0J 8 0.06 0.00 7.89 0.37 0.67 0.03 032 0.00 0.12 0.00 0.02 (2.09) 0.00 0.15 0.00 0.07 0.20 1.21 0J3 2.92 0.00 0.38 0J 0 0.18 0.00 0.32 0.33 0.00 0.00 8.07 0.54 0.52 00 0.27 0.03 0.03 0.00 0.17 (1.02) 0.03 0.07 0.13 0.07 0J4 0.78 0.40 0.91 0.00 0.24 0.20 0.07 0.24 0.51 020 0.07 0.03 5.85 (Underreported-*- + + ) for mother's age (Including TEF) (AH types) (belated, ulnar) This study provides data from the following countries of SEA: 1) Hong-Koag: 10.001 total deliveries; 2) India: 39.438 total deliveries horn Csharnn. and Bombay, respectively; 3) Malaysia: 36,123 total deliveries from Kuala Lumpur, and Singapore, respectively; 4) Philippines: 29,989 total deliveries from Mantle. There were 155,573 total deliveries (ie, single and multiple). The sampling of data was organized in 1962 and 1963. respectively. 0010816 ' ^ 7 - * H r7 o i irigin. :ts has mpor* a rio ^ 'th e : sor^e ! eartiL n a lfo rpp. 1- +++) ge ) ) Bombay, jv 4) H erbicide Exposure in Vietnam 235 Hydaddiform mole is a distinct clinical entity, normally requiring hospitaliza tion. It is therefore assumed that data currently being collected in hospital settings provide reasonably complete, accurate estimates o f its occurrence. A Word is in order as to the reliability o f historical hospital statistics in Vietnam as compared with those derived from study of the original health records. At least one author, D r. Nguyen Thi Ngoc Phuong [14], states in her paper that government statistics, at least until recently, are unreliable. One o f us (J.D .C .) was personally involved in studying some o f these for the period from 1966 to 1972 [IS ]. The relation between midwives* records and figures reported by the Ministry o f Health is very poor. A report by Cutting et al for the U.S. Department of Defense [16], described subsequently, is significandy flawed in this respect. For example, figures obtained from the original midwife records in Tay Ninh are at striking variance with those recorded by the Ministry o f Health and subsequently reported by Cutting. Control fo r Potentially Confounding Variables Only limited efforts have been made in these studies to consider the potentially confounding effects o f extraneous variables (eg, maternal age, nutridon, infection) which might distort any observed associadon between herbicide exposure and repro ductive outcome. Variables which have been controlled, either in design or analysis, w ill be mentioned in summarizing the individual studies. DESCRIPTION OF THE VIETNAMESE STUDIES We shall now consider the nine Vietnamese investigations which were presented to us.* To help the reader follow the ensuing discussion. Table n lists each study in turn, together with a brief description o f the research design. Studies Carried O ut in the South o f Vietnam Khoa. D r. Nguyen Dinh Khoa o f the University o f Hanoi presented a paper entitled "Some Biologic Parameters Collected on the Groups o f People in an Area Affected by Chemicals," [12]. D r. Khoa collected retrospective histories covering the years between 1965 and 1982 from 313 families o f Montagnards living in an area o f heavy spraying. In this one paper the original figures as presented appeared to us so confusing that we have had to extrapolate and summarize them. Infant mortality was reported to be 14.7% (176 death/1,196 births) and the rate o f miscarriage was 10.1% (134/1,196 + 134). The incidence o f birth defects was "To property appreciate the Working Group'* evaluation of these paper*, we should note some of the practical considerations that controlled our discussion. By an heroic effort on the part of the Vietnamese participants, all of the papers had been translated into English and distributed to all Congress participants at the time of registration. The members could, therefore, study the papets before their presentation in the venous working groups. In some cases the authors had not been able to review folly these English translations. In other cases, either immediately after the initial presentation in English. French, or Vietnamese or on sulwrquent days in answer to questions posed previously, the numbers in the reports as we individually received them were corrected, emendated, or significantly augmented. In one paper, for example, all of the controls were accidentally omitted from the English version. The constraints of time, and sometimes of language, prevented these additions from being fully incorporated into bur discussion and statistical evaluation of these papers. Most of our comments, therefore, are principally based, though perhaps in some rases unfairly, on the English version after the correction of some obvious errors, not upon the "final" and complete Vietnamese paper. French provided the common language for many of the questions and much of the diMiiwin of the various repons. 4875 0010817 90H2IS9W I 236 Constable and Hatch TABLE H . Vietnamese Studies of the Reproductive Effects of Herbicides Authors Site, population studied Research desien 1. Khoa. 1983 [12] 2. Nguyen. 1983 [13] 3. Tnntg and Chien. 1983 [17] 4. Huong and Phuong, 1983 [18] 3. Phuong and Huong, 1983 [14] 6. Lang, 11119, and Van. 1983a [10] 7. Lang, Van. Dwyer, et ai. 1983 [11] 8. Can. Xiem. Hong, et al. 1983(8] 9. Can. Xiem. Tong, et al, 1983(9] Montagnards living in a heavily sprayed area of Southern Vietnam Provincial hospital in a heavily sprayed area of Southern Vietnam Families from a sprayed and an unsprayed village. Southern Vietnam Obstetric Hospital. Ho Chi Minh City (Saigon), Southern Vietnam Women from a sprayed area of Southern Vietnam and from un sprayed areas of the South and the North Residents of agricultural villages for veterans. Northern Vietnam Families of veterans. Northern Vietnam Veterans living in three areas of Northern Vietnam Malformed and oormai offspring of veterans living in Northern Vietnam Rates of obstetric events for s pe riod during and after the spray ing. 1963-1982. Raws of obstetric evens. 19791981. Comparison of rates of miscar riage and birth defects before and after spraying in an exposed and unexposed village Time trends analysis of reproduc tive events. 1932-1981. Casecontrol study of herbicide expo sure and hydaodiform mole Comparison of reproductive prob lems in women exposed and unexposed to herbicides Comparison of birth defects in offspring of soldiers who did aad did not serve in the South Comparison of miscarriage rates according to degree of hus band's herbicide exposure Comparison of reproductive evens among wives of exposed and tmexposed veterans Case-control study of bitth defects in relation to father's service in the South ____________ __ 2.7% (33/1,196), but it is unclear how many cases o f infantile paralysis may have been included amongst the congenital anomalies. Although unfortunately no control data were included in the paper as translated into English, these had in fact been collected in an unsprayed village; the controls demonstrated a miscarriage rate o f 6.1% (38/387 + 38) and a 1.0% rate o f congenital anomaly (6/387), both rates being lower than the comparable occurrence in exposed villages. No statistical evaluation o f these "differences'* could be attempted by the Working Group since the control figures were not available in the original paper. Nguyen. D r. Ho Dang Nguyen o f the Tay Ninh Polyclinic Hospital presented a paper entitled "Pregnancies at the Polyclinic o f Tay Ninh Province,1' [13] summariz ing the results o f a study o f reproductive outcomes occurring between 1979 and 1981 in a provincial hospital serving an area heavily sprayed until about 1970. Although the province has a total population o f some 700,000 living in seven districts, the Tay Ninh City Polyclinic cares primarily for a population o f about 30,000 and in addition receives many difficult cases referred from the district dispensaries which handle most deliveries. Combining the figures from 1979, 1980, and 1981 and the first 6 months o f 1982, there were 7,344 deliveries, resulting in 166 stillbirths (2.3% ); 1,633 premature births (22.2% ); 133 hydatidiform moles (1.8% ); and 78 congenital defects V - H CI 7 9 - 0010818 < 4876 |8 *6 S U *0 (1 c ; >r a pespny9 7 9 -^ :forc " " reposed ,,. rodoe*-.- Zase-x^. : expo-. >ie : prob ind m did xith raies efects ce in have ntrol been ic of eing ition itroi ed a iriz981 ugh Tay cion idle st 6 633 sets Herbicide Exposure in Vietnam 237 (1.1% ), while 1,920 abortions were recorded out o f a total of 9,264 pregnancies (20.7% ). To the 78 congenital defects noted from this part o f the study, 37 defects referred in from the districts were added, presumably only a small proportion o f those actually bom. O f the 37 referred mothers, one had had malaria and two had positive tests for syphilis. Smoking and intrapregnancy use o f medications were very limited. The mothers o f the 37 had had 202 total pregnancies terminating in 20 abortions, 4 premature births, 32 stillbirths, and 126 fullterm babies. Among the 37 defects, two were monsters, eight showed anencephaly, four showed anopthalmia. four showed phocomelia, and four others showed severe skeletal deformities; these were anomalies referred to the polyclinic rather than representing an unselected sample o f all congen ital defects in the area. There is some ambiguity in the paper in that the 37 were "sent from the district" as distinguished from the 78 bora at the polyclinic, but nonetheless among the six mothers (o f the 37) for whom the location o f the delivery is given, three were, in fact, delivered at the polyclinic. Note was also made o f two mothers, each o f whom delivered four children with deft lips and another who delivered four blind children. These last are in addition to the four previously noted cases of anopthalmia (ie, not included in the 57) even though the children were apparently not bora at the polyclinic and might normally have been considered as "sent from the district." This investigation is one o f several presented to the working group which, while not susceptible o f statistical analysis, seem to show a large number o f striking and usually rare anomalies, even allowing for their being selected. Thin g and C hien. D r. Cung Binh Trung and Nguyen Tran Chien o f the Medical College o f Hanoi presented a paper entitled "Spontaneous Abortions and Birth Defects in Area Exposed to Toxic Chemical Sprays in Giong Trom District, Ben Tre Province, South Vietnam" [17]. They studied the occurrence o f spontaneous abortion and birth defects before and after the time o f spraying. The authors surveyed 848 families who continued to live in heavily sprayed areas; families living in a nearby unsprayed village served as a comparison group. The method o f selecting study participants is not described. Before the spraying, the rate o f miscarriage in these 848 families was 5.6% (129 miscarriages/2,292 pregnancies); after the spraying the rate was 13.9% (217/1,562). The control families reported a miscarriage rate o f 7.3% (32/436) prior to the time o f spraying and 7.4% (27/365) after the time of spraying. This being a pre/post exposure comparison o f the same fam ilies, the mothers were likely to be older in the postexposure period and on this basis alone might have been expected to show some increase, even in the unexposed controls. Birth defects (apparently corrected by exclusion o f cases induced by drugs or diseases such as syphilis) were also studied in the exposed villages, though not among the controls. Prior to the spraying, the rate o f defects among surviving live births was 0.14% (3/2J63 births) compared with a rate o f 1.78% (24/1.345) among children bora after the spraying. The preexposure rate seems to reflect very limited reporting and again mothers were older in the period after exposure. O f the 24 anomalies occurring in the postexposure period, four were cases o f deafness and eight involved skeletal defects. Huong and Phuong. Drs. Le Thi Diem Huong and Nguyen Thi Ngoc Phuong o f the former Tu Du Gynecological and Surgical Hospital in Ho Chi Minh City presented two papers. The first o f these, entitled "The State o f Abnormal Pregnancies and Congenital Malformation at the Gyneco-Obstetrical Hospital o f Ho Chi Minh 0010819 \ ( 4877 |8 1 6 8 U R O d 238 Constable and Hatch City, formerly Tu Du Hospital," [18] was divided into two sections. The first section reports on time trends in the incidence o f abortion, "intrauterine deaths" (? still births), molar pregnancies, and congenital anomalies between 1952 and 1981, though the figures are unavailable for some years in each category (see Table ED following). Figures for the years prior to 1975 are drawn from doctoral theses, since administra tive data for this period are viewed by the authors as unreliable; figures for the years since 1975 come from the annual report of the hospital. The authors recognize that there is some element o f error in that the denominator--total number o f pregnancies-- is not always consistent. Although striking changes in rates are demonstrable, interpretation is somewhat difficult; however, the authors feel that the data, even though provided by different sources, are comparable. The abortion rate appears to remain fairly low until 1967 (the rate reported for 1952 seems contrary to general experience), then rises dramati cally until 1978 and then declines slightly. These changes could be considered to be consistent with the times of heaviest spraying if a persistent effect is assumed. Stillbirths appear to rise between 1952 and 1953 (but again the figures seem very low TABLE m . Data From Hnong and Ptmong, 1983 {181 Years Molar pregnancies and choriocarcinoma (X ) Congenital malformations (X ) 1952 56/6,495 1953 0.78 1959 1960 1962 1963 1964 1965 173/ 14,413 160/ 14.076 158/ 12.440 1.20 1.13 1.27 93/12,538 106/16.779 96/18.463 0.73 0.63 0_52 1966 82/19.429 0.42 1967 1971 1976 1977 1978 1979 1980 1981 350/ 24,345 242/ 27.457 338/ 16.167 499/ 12.943 477/ 13,544 580/ 12.757 460/ 12.766 569/ 12.754 1.43 0.87 2.09 3.85 3-52 4.54 3.60 < 4.19 128/23-509 O S I 92/12.796 0.70 101/13.430 0.75 134/12.648 1.06 158/12J73 1.24 133/13.430 0.99 Imra-uterine deaths (X ) 38/6.495 0.58 10/6.889 0.12 Abortions (X ) 29/6.495 103/ 6.889 0.45 1.20 916/ 19,308 820/ 19.854 378/24-345 1-56 3.594/ 24-345 382/27,475 1.39 3.822/ 27.475 216/16.167 1.33 2.732/ 16,167 232/12,943 1.78 2.139/ 12.943 196/13,544 1.44 2.458/ 13-544 ; 187/12.757 1.47 1,396/ 12.757 185/12,766 1.45 .1.624/ 12.765 239/13-574 1.76 1.367/ 13-574 4.73 4.13 14.58 13.99 16.89 16-52 18.14 10.94 12.73 10.09 001082 o;?re46SI2NOn fi ar cs cc tu rr dr D a. w (c ir ft li: cr 8' b a ir c: ti ll a c E section ? still-- though twing). inistrae yeSrs ze dt icie st r.^ newhat fferent il 1967 ramatid to be sumed. try low nions (%) Ve 0.45 1.20 4.73 4.13 14.58 13.99 16.89 16.52 10.94 12.73 10.09 Herbicide Exposure in Vietnam 239 and only 2 years are available) and increase again in 1967, from which time they seem to remain fairly constant until 1981. Again these changes could be consistent with a herbicide exposure if one assumes a persistent effect. Moles and choriocarci nomas are among the most complete figures available and again show a sharp increase,'but this is first apparent in 1976 (1971 available, but not 1972-1975), well after heavy spraying had ceased. Congenital anomalies, presumably restricted to grossly recognizable external defects, show no great changes and only a slight increase in 1979, 1980, and 1981 as compared to 1977,1978, and the preceding years. In considering these figures, it should be remembered that Tu Du is a referral hospital and the rates o f some reproductive problems may be somewhat higher than for Ho Chi Minh City as a whole. The second section o f this paper reports a case-control study comparing the frequency o f herbicide exposure among 10 0 women with molar pregnancies (cases) and 284 women with normal deliveries (controls). Control mothers were matched to cases on maternal age and parity; however, the groups differ significantly in social conditions and diet (8 6 % o f the women with molar pregnancies were considered to have a "good" living standard against only 45 % o f the controls). Moreover, strikingly more o f the husbands o f women with molar pregnancies smoked (84 vs 70% ) or drank alcohol (90 vs 35% ), although it is not clear that these are relevant differences. Data on past exposure to herbicides were collected at interview. O f the 85 out o f 100 cases whose exposure status was ascertained, 48 or 56.5% were considered exposed, while o f the 276 controls for whom determination o f exposure was made, about 27 (out o f 284) or 9.8% reported exposure. (No reason is given for the lack o f exposure information on 15 cases and eight controls.) The data are summarized below in a fourfold table (Table IV ) to which we have added the odds ratio and 95% confidence lim its. The data suggest a strong association between herbicide exposure prior to conception and subsequent development o f a hydatidiform mole. M olar pregnancies, generally more common among Asian women, are conceptions without an embryo, but with grossly swollen chorionic v illi. Complete moles are androgenetic in origin, arising from fertilization o f an ovum in which the nucleus is either absent or inactivated [19]. M olar pregnancy is often associated with subsequent development of choriocarcinoma. Tissue determination o f dioxin levels in the fat or other tissues of the patient would be very helpful in confirming the possible association between herbicides and hydatidiform mole/choriocarcinoma. In this study, data on congenital anomalies have also been gathered, but the figures are too small as yet to draw any conclusions; fortunately, the authors plan to expand their research. Phuong and Huong. The same authors presented another paper entitled "The Effects o f Toxic Chemicals on the Pregnancy o f the Women Living at Two Localities TABLE IV . D ata From H uoog and P b aou f, 1983 ra r__________________________ ** Expowd Not expowd Caw 48 f Control 27 Total 93 37 249 286 OR - 11.96(6.67.21.46). 4879 0010821 M B I ZMOtl DON2 159485 2 4 0 Constable and Hatch in the South o f Vietnam'' [14], This inquiry presents a comparison of reproductive problems reported by the women from a sprayed village, in which all were considered exposed, with those recorded from a group o f women in Ho Chi Minh City, amongst whom 92% were considered unexposed. The time period for these reproductive histories and the method o f selecting subjects for interview are not described. The tables showing the reported results are appended here (Tables V ,V I). The original paper reported a comparison by number o f families interviewed: 1,249 and 1,244, respectively, the latter figure comprised o f 1,126 unexposed and 98 exposed women. (These are the figures used in the original tables but they add up to 1,224 not 1,244.) A comparison o f total pregnancies was subsequently added and percentages deter mined from these; 98 exposed families in Ho Chi Minh City were excluded from these calculations. A further comparison is then made between the figures for the exposed population and those from a number o f villages in North Vietnam that show considerably lower rates (Table V I). The authors state that the number o f reproductive abnormalities varies in a significant manner with respect to the degree o f exposure, but this statement is not further elucidated in the paper. In contrast to their previous study, the authors give no information as to the changes in successive years nor as to the rates before spraying. Note that the rate o f molar pregnancy among the exposed mothers is 1.8% (133/7,327). W hile considerably higher than the rate o f 0.4% in the unexposed comparison group, it is only slightly higher than that o f 1.2 0 % reported in the previous paper from Tu Du for 1959 to 1962 [18]; ie, before significant spraying, and lower than the rate o f 3 to4% observed at Tu Du since spraying. Again, however,, the comparison with Tu Du may not be legitimate in that complicated pregnancies are likely to have been referred there. The rate for abortion at 8 % is between that for Tu TABLE V . The State of h th o io g ia l Pregnancies at the Tbang Phong Village (Ben tre) and at the 11 10th District of Ho Chi Minh City [14]_______ Congenital anomalies Embryo death in wens Natural abortion Molar pregnancy Prenatal death of newborn Thang Phong village: exposed group, 7,327 pregnancies (% ) 81 ( 1. 1) . 39 (0.8) 387 (8.0) 133 (0.7) 924 (12.4) 10th distra of Ho Chi Minh dry: nonexposed group. 6,690 pregnancies (*) 29(0.4) 2 (0.0) 243(3.6) 26 (0.4) 1 (4.6) TABLE V I. Data From Phnom and Huong, 1983 {141 South of Vietnam - Thanh Phong (% ) My Van Birth defects Embryo dead) in mens Natural abortion Molar pregnancy 1.1 0.43 0.06 0.8 1.91 0.13 8.01 3.77 0.21 0.73 0.09 0.04 North o f Vietnam HaiHau 0.39 0.03 1.91 0.12 4.96 0.018 0.03 0.01 Mai Chau 0.68 0.12 3.85 0.29 8.74 0.040 0.10 0.13 4880 0010822 .:ive sred ngsi .live T h e -linai-- 244-g . nen,ur 44.). ter-^v.: Tom ' th* how*^ nive A in, ious is to osed i the si in in g, rvcr. i _j die c ity : up. .it :2 :9 MO 3 Herbicide Exposure in Vietnam 241 Du 1965-66 (4.5% ) and 1967-68 (an average of 16%). That for stillbirths at 0.8% is about half the Tu Du rate for 1967--81 (average 1.5%) and that for congenital anomalies at 1.1 % is about double that of Tu Du for 1963-67 and comparable to 1979 and 1980. always remembering that most of the Tu Du population was not severely exposed. Studies Carried Out in the North of Vietnam Lang, Tung, and Van. We come now to a consideration o f those two Vietnam ese investigations that at the time o f their presentation seemed to be the most complete, and which concern themselves with die possible deleterious effects o f male herbicide exposure. The first is that of Drs. Ton Due Lang, Ton That Tung, and Do Due Van from Viet Due Hospital in Hanoi. " Mutagenic Effects on the First Genera tion After Exposure to Agent Orange" [10]. This work is a continuation o f the seminal investigation originated by Dr. Ton That Tung, the first results o f which were made available to the West, albeit in a very limited way, through his manuscript " Le Probleme des Effets Mutagenes sur la Premiere Generation apres L 'Exposition aux Herbicides" early in 1980. These figures were augmented and some other changes made by D r. Ton That Tung in a second manuscript in 1981 and are further supple mented in the repon we are discussing now. The following pages w ill synthesize these three sources o f information, though only the last was discussed and evaluated at the conference. The investigation consists o f an epidemiological survey, an analysis o f the nature o f the congenital anomalies encountered, and a correlation o f the degree o f herbicidal exposure with the rate o f occurrence o f congenital anomalies. The epidemiological survey was carried out in two steps: initially by studying the obstetrical statistics of veterans' wives in two villages, one containing many veterans returning from the south, and one not. This was subsequently extended to an investigation carried out at a number o f agricuiatural cooperatives set up principally for veterans. The husbands were divided into those who had served in the southern part o f Vietnam, all o f whom are assumed to be exposed (although most were Northerners returning home, the manuscript o f Ton H o t Tung records that some may have been Southerners now going north for the first time), and those who had remained at home and were consequently unexposed. Ton That Tung states that ail o f the exposed fathers o f the anomalous children told him o f being directly exposed to spraying and that their average stay in the South was from 3 to 4 years. Marriages for exposed men normally occurred after their definitive return following at least a brief period o f courtship. Therefore the delay between possible exposure and conception w ill have been more than the 80 days that sperm normally survive. The results o f the study appear in Table V II which shows that half the congenital anomalies found at Yen Bai were bom to exposed fathers, though these made up only some 16.6% o f the potential breeding male population (Ton That Tung: 700 out o f 4,200) and in Quy Mong all the anomalies were bom to exposed fathers, even though they made up only 10% o f the potential breeding male population (using the same proportions as determined for Yen Bai. since no precise figures are given). There is no explanation offered as to the cause o f the somewhat higher birth rate in Yen Bai as compared to that in Quy Mong, even allowing that the figures for Yen Bai include 4 years and those o f Quy Mong only 3. A ll 30 congenital anomalies encountered at Yen Bai are then listed and include six anencephaiics o f various kinds as well as two major limb deformities, all amongst 4881 0010823 p - W `i'7'7 886SIZN0Q 016SIZHOO 242 Constable and Hatch TABLE V P . Congenital Malformations in Obstetrical S a r t o (10] Yen Bai Local population Local demobbed servicemen Time of statistics Number of binhs Number of congenital malformations Congenital malformations of civilian population Congenital malformations of veteran's children 35.000* 700* 1975-1978 3.058e 30* IS 15 *32,000 (Ton That Tung #1). *"Approximately" 700. *3,058: 311 birth* to exposed: 2.347 births to unexposed *of the 30:22 at term: eight premature. *Diviskm of births unknown: he guesses 90 and 143. Quy Mong 4,500 30 1976-1978 233* 9 0 9 children of exposed fathers. At Quy Mong they reported three anencephaiics, all to one pair of parents, and three major deformities, ail with fathers exposed (vide supra). It should be noted again that many o f these details are taken from the manuscript of D r. Ton That Tung, not horn the abbreviated text as presented to the conference, though o f course this includes all o f Ton That Tung's cases in the total figures reported. In the agricultural cooperatives, which were largely composed o f veterans from the South, a total o f 1,142 exposed veterans produced 3,147 children with 71 anom alies (2.25% ) while 613 unexposed married veterans had 2,172 children with 10 anomalies (0.46% ). a difference o f fivefold. Among the 71 anomalies born to exposed fathers were six anencephaiics, one meningocele, two anopthalmias, three major limb deformities, and one amorphous monster. Below we have abstracted data from Ton Due Lang's paper to compare against the data from D r. Ton That Tung's manuscripts (Table V ID ). Ton That Tung's series consists o f only 47 anomalies which are presumably included in the final count o f 71; hence there would appear to be some discrepancy in the definition o f congenital heart anomalies. Table IX shows Lang's data on the frequency o f reproductive outcomes by gravidity. The wives o f unexposed fathers demonstrate the increase in the frequency o f adverse outcomes with successive pregnancies normally expected, whereas this trend is reversed among the women whose husbands were exposed. The pattern is suggestive o f a toxic effect most virulent at the time o f first conception and then gradually diminishing in its potency. The paper concludes with a map showing an increased percentage o f anomalies among soldiers serving in particularly heavily defoliated areas, but no specific figures or controls are provided. It is o f interest that in Ton That Tung's original manuscripts he describes for the same group o f cooperative veterans a striking change in reproductive endpoints other than congenital anomalies, as shown in the appended table (Table X ). This was not included in the paper presented at the conference. D r. Tung in both his manuscripts notes apparent increases in hydaddiform moles in Hanoi. He stares that at the moment o f his writing there were 19 cases o f hydatidiform mole in his hospital, nine o f them in wives o f exposed soldiers, but no other figures are given for this anecdotal report. 0010824 4 i to -a), of ce. res Dm p* ed nb bn DCS ire ne y =y iis is Ml in iy ie :r Dt 71 if O Herbicide Exposure in Vietnam TABLE V ili. Comparison of Figures From Tung and From Lang_____________________________________________ Ton That Tung (H) o u to fm aio f47 Lang out of total of 71 Anenceptuly Anopdiaimia Cleft Up and pelare Polydactyly Congenital been Hemangxxna 6 1 4 4 18 0 6 2 9 7 11 (Sic) 4 243 TABLE IX . Frequency of Reproductive Incidents According to the Ordinai Number of Pregnancies [101___________________ Ofdinal number of pregirency Affected group (*) Nonaffecred (*) 1 20.44 3.42 2 13.64 3.98 3 13.37 7.88 4 13.22 8.76 3 13.93 9.74 6 13.00 13.00 7 7.60 19.38 TABLE X Investigation o f 1,549 Vietnamese Soldiers* Group Congenital malformations Number % Abonions Number % Premature deliveries Number % Sterile Number % A 47 3.14 232 14.42 30 2.01 22 2.8 (out of 1,496 (out of 1.748 (out of (out of births) pregnancies) 1.496 786 deliveries) couples) B3 0.21 143 9.04 9 0.61 3 1.2 (out o f 1.438 (out of 1.381 (out of (out of births) pregnancies) 1.438 418 deliveries) couples) p 1. 10"' p I. I0 *s p 1. I0~ J p0.03 Data from Ton That Tung, "La problme des effets mutagenes sur la premiere generation aptes l'exposition aux herbicides." Group A; 936 individuals with 786 couples in which the husband lived in the sprayed iones of South Vietnam: group B; 393 individuis with 418 couples in which the husband was not in South Vietnam (control group). Lang, Van, Dw yer, et ai [11]. An important addendum to this study was provided in a paper entitled "Self Reports o f Exposure to Herbicides and Health Problems--A Preliminary Analysis o f Survey Data From the Families o f432 Veterans o f Northern Vietnam" [11] authored by D r. Ton Due Lang and others and presented by one o fthe authors. D r. James Dwyer o f the State University o f New York at Stony Brook. This preliminary investigation describes a survey o f432 veterans from whom a medical/reproducdve history was obtained by one interview team while a separate team, blind to the first, determined presumptive herbicide exposure. This included, very significantly, an estimate o f the degree o f exposure as indicated by whether the 4883 0010825 BON 2 1 5 9 1 8 8 D 0 H 2 1HH-8 9- 2 4 4 Constable and Hatch individual 1) sustained direct or wet exposure, 2 ) lived in a defoliated area, or 3) only passed through a defoliated region. Coupled with a history o f military service between 1964 and 1970, this allowed division o f these respondents into high, moderate, and low exposure subgroups. The sample (N * 432) is too small as yet to provide data on rare events such as congenital anomalies and in terms o f reproductive outcomes only miscarriages were analyzed. The number o f miscarriages was ascertained in two ways: 1) The wife was asked to report the number o f live births, miscarriages, and stillbirths that occurred prior to, during, and after her husband's military service and 2 ) a complete reproductive history was obtained in which pregnancies were reported by date of delivery or other termination. These two methods o f obtaining what should have been the same information showed surprising disparities and their inconsistency is evidence o f limited reliability. However, the general pattern o f events, especially after the father's time o f military service, was the same by either method o f inquiry. For younger mothers, there was no apparent association between the degree of exposure of her mate and her risk of miscarriage, and for all age groups combined, the rates were within the range o f freguency found in other Vietnamese study populations. However, for older women there was shown to be a strong association between the extent o f paternal exposure and the rate o f miscarriage. These figures are not fully convincing since the reported rates of miscarriage among the unexposed older mothers (0 out o f 60 by one method o f inquiry; 3 out o f 6 6 by another) are too low to be fully credible. Also it is not immediately clear why older women would be more affected by their husband's herbicide exposure than would be those who were younger. Furhermore, as noted previously, if women o f all ages are combined, the association largely disappears. Can, Xiem , Hong, et a l. We now come to the final Vietnamese report which in consideration o f adherence to an appropriate protocol, completeness o f figures, and the possibility o f precise statistical analysis is the most convincing o f those presented to us. D r. Nguyen Can and his co-workers from the Institute for the Protection o f Mothers and Newborn in Hanoi presented "An Epidemiological Survey o f Pregnancies in North Vietnam" [8 ] together with a very important addendum. "A Case Control Survey o f Congenital Defects in M y Van District, Hai Hung Province" [9]. As in the previously discussed study o f D r. Ton Due Lang [10], a survey was made o f the rate o f adverse reproductive outcomes among the wives o f 40,064 veterans exposed or unexposed to herbicides. Three areas in North Vietnam (ie, unsprayed) were arbitrarily selected--one in the mountains, one in the lowlands, and one on the coast. The women were 90% rice growers (the remainder medical staff, office workers, etc.) and are reported to have had no history o f tuberculosis, syphilis, or malaria nor to have had drug exposure during pregnancy. It is not dear whether there were, in feet, no women found to be so afflicted or whether the investigations exduded those that were, since 40,000 women free o f any such disease is hard to credit. A ll 40,064 interviews were carried out in 3 months (June-August 1982) using three doctors, thirty midwives, and additional ancillary personnel--a truly formidable undertaking. An effort was made to validate all reported reproductive events. Al though questioning was concentrated on co-ops and production teams to increase the yidd o f exposed fathers (11,053 or 25% ), nonetheless all married couples p resen t in the locality at the time o f the interview were included and it was estimated that 7 0 90% o f those domiciled were questioned. A:r. QU Q A'-C OC1 5 8 2 6 'jJ - W O / nly ;en in d ata nes wo in d in d -- le d u ld " ' ncy^ illy iry- r_- O fcj ed. idy . ion are sed too i be ere the rt>. ose the vey -A :e~ vas )64 lie . rnd iff. lis. her jns i to ing bie A lthe r in 70- Herbicide Exposure in Vietnam 245 The results o f these interviews are shown in Tables X I and X II. The data appear to show a small increase o f borderline statistical significance (a * . 10. two-tailed) in the rate o f miscarriages and congenital defects among women whose mates were exposed, but no increase in molar pregnancy or stillbirths. Once again, the total rate o f congenital anomalies is very low, even in the exposed group (0.64% ). The authors themselves-point out, however, that no cardiac defects or hemangiomas are listed. Presumably most o f the defects reported are grossly detectable abnormalities. It is also noted that the exposed group have fewer pregnancies on the average than did the unexposed (2.91 vs. 4.24) and that maternal age was not controlled, although it was in the case control study soon to be discussed. There is no precise breakdown as to occupation nor a distinction made between soldiers that served as officers, but there is good evidence that, at least in the countryside o f North Vietnam, the general conditions o f life vary iitde with these parameters. An analysis of the distribution of specific congenital anomalies is then given. This seems to show a slight excess among exposed fathers in the proportion o f anencephalics (6.3% vs. 4.6% ). A comparable proportional increase in deft lip and palate is also shown for exposed fathers whereas in contrast to the figures in previously discussed reports, limb deformities are more frequent among children o f unexposed fathers. Can, Xiem , Tong, et al [9]. A case-control study was set up by the authors to answer some o f the possible objections to the investigation previously concluded. The districts for follow-up were drawn by lot. Sixty-one case families were chosen, each with a surviving child with a congenital anomaly--d eft palate, deft lip, limb malfor mations, megacolon, absence o f the ears, imperforate anus*, congenital cataracts, or congenital blindness. O f these 61 anomalies, 30 were in the children o f exposed fathers (49.2% ). Three controls were drawn by lot for each o f these 61 cases from a pool matched to the cases on maternal age (plus or minus 3 years); number of deliveries (plus or minus 2); village o f residence: and age o f a living child (plus or minus 2 years). Among the 183 control couples, 39 fathers (21.3% ) were exposed. Almost all subjects were currently ricefield workers. None had a history o f heart disease, V D , cancer, malaria, etc. None gave a history o f addiction to smoking or alcohol. Most o f the women were less than 36 years old (31 out of 61) and the controls and cases were well-matched as to age. The investigators calculated odds ratios and 93% confidence intervals for the association o f interest. As can be seen in Table X m , the risk o f having served in the South is about 3.3 time greater for fathers o f cases than for fathers o f controls. Stratifying on maternal age ( < 3 6 , ^ 3 6 ) produces similiar results and no evidence o f an interaction with age o f the mother. Providing that this study was indeed carried out as described, then it shows a statistically significant association between paternal "exposure'' to herbicides and certain congenital anomalies in the offspring. SUMMARY OF RESULTS FROM TH E VIETNAM ESE STUDIES The studies carried out in the North o f Vietnam are consistent in reporting an association between presumptive paternal exposure to herbicides prior to or at con vince unoperated imperforate anus is fatal, we are a little uncertain as the true nature of the anomaly meant herte. 4 B S I- 0010827 I iim izMoi VK tt.. TABLE X I, P r ti From Cnn et al, 1W3 |g|__________________________________________________________________________ Group Number of women investtested Number of pregnancies Number of deliveries Number of bodions Number of cordages Number of molar pregnancies Dead fetuses Cues with congenital defects A(unexposed) B(exposed) Both (roups 29.041 11,023 40,064 121.993 32,069 134,062 114,023 29,360 143,383 7,148 2,271 9,419 730 210 960 70 2.312 321 28 376 189 98 3,088 712 i 4886 0010828 i I 6 1 6 S lMd i * -- * ** Herbicide Exposure in Vietnam 247 TABLE XU . Statistics of Abnormal Pregnancies in Women of A and B Groups in Three North Vietnamese Districts [81______________________________________________________________________________ Group Abortion/ ' ( * 2 SD) Molar pregnancies/ No. of pregnancies (% 2 SD) Fetuses dead before and during delivery/ No. of deliveries (% 2 SD) Fetuses with congenial defects/ No. of deliveries (* 2SD) A(unexposed) B (exposed) Both groups 7.146/ 121.933 (5.16 0.13) 2J74/ 32.069 (7.08 0.28) 9.419/ 154.062 (6.11 0. 12) 70/ 121.993 (0.06 0.01) 28/ 32.069 <0.09 0.04) 98/ 154,062 (0.06 0.01) 2,512/ 114.025 <2J0 0 .0 9 ) 576/ 29,560 (1.95 0.16) 3,088/ 143,583 (1.15 0.07) 521/ 114,025 (0.46 0.04) 188/ 29,560 (0.64 0.09) 710/ 143,585 (0.49 0.03) TABLE X m . Dam From Can e ta L P O P l* Not exposed Cases Controls 30 39 37 144 OR - 3J7 (1.91.6.69). ception and congenital defects in subsequent offspring, particularly certain types of anomaly (anencepbaly, orofacial defects). The strength o f the association demon strated in the data varies among studies. Results in respect to miscarriage are conflicting. No association with molar pregnancy is documented. The studies carried out in the South o f Vietnam, where women as well as men were at risk o f exposure, report increases in miscarriage, stillbirths, molar pregnancy, and birth defects, again with certain types predominating, among couples previously exposed to herbicides. In the case o f molar pregnancy the evidence for an association with herbicide exposure is very suggestive. OTHER STUDIES OF H E R B iaO E EXPOSURE IN VIETNAM For purposes o f completeness and comparison with the results o f the Vietnamese studies, we briefly describe the other investigations pertaining to herbicide exposure in Vietnam and possible reproductive damage. These include the following: two U.S. studies o f South Vietnamese women [16,20]; an Australian Government study of birth defects in relation to father's Vietnam service [21]; a study o f A ir force personnel associated with Ranch Hand, the U .S . Government's herbicide-spraying operation [7]; and the Centers for Disease Control study in Atlanta o f birth defects. Vietnam service, and potential Agent Orange exposure [5,6]. Studies o f those exposed to herbicides or dioxins in other circumstances (eg, Seveso, herbicide production facilities, contaminated residential sites) w ill not be discussed here. 4887 0010829 18468 IZM0I 2 4 8 Constable and Hatch U .S . Studies o f South Vietnam ese Women The study mentioned above by Cutting et al [16] for the Department of Defense utilized obstetric data from the records o f selected hospitals in Vietnam to compare the rates of stillbirth, hydatidiform moie, and congenital defects in periods o f light (1960-65) and heavy (1966-69) herbicide spraying. In the Coastal Plain and Delta areas o f South Vietnam, rates rose in the heavy spray period but elsewhere rates were constant or declined. As discussed above, this study is flawed because o f inadequacies in the data base. A study by Kunstadter [20] for the National Academy of Science used the obstetric records of selected hospitals in Ho Chi Minh City (Saigon), supplemented by personal interviews, to gather information on birth defects and perinatal mortality. Maternal herbicide exposure was established based on information contained in the Defense Department's HERBS tapes. The data showed no consistent association between congenital malformation and maternal exposure in the first trimester. Cleft lip, however, increased relative to other defects during the heavy spraying period and remained elevated. The stillbirth rate rose, but not until after spraying ceased. Australian Study of Birth D efects and F ather's Vietnam Service In January 1983 the Australian government published results o f a large casecontrol study o f some 8,500 children with birth defects (largely structural defects detectable at birth) and 8,500 livebom controls, matched for maternal age and time o f birth [21]. Cases with no father's name included on the birth certificate were excluded from the study. The frequency o f Vietnam service among the fathers, as determined from Army lists, was virtually the same in cases as in controls. In relation to specific birth defects the report states that the odds ratios for Down's syndrome and ventricular septal defects "exceed 1-5, the minimum odds ratio that can be usefully studied in an epidemiological study," but these data are not shown. No index o f exposure o f Australian troops to herbicides could be developed. Thus the results of this study apply only to service in Vietnam and not to potential effects o f herbicide exposure sustained there. In feet the investigators state that "exposure to herbicides was infrequent and probably very low in Australian troops in V ietnam ." A ir Force Study of M orbidity in Ranch Handers In February 1984 the A ir Force released its preliminary study o f morbidity in Ranch Hand personnel, the approximately 1,200 men who conducted the herbicide spraying missions in Vietnam and who some maintain sustained the highest exposures o f any American troops [7]. (Others argue that ground troops living in contaminated areas, without opportunities to shower and change clothes, may have received a dose as high or higher.) An exposure index was calculated based on the amount o f herbicide used during the Ranch Hander's tour o f duty. Ranch Handers were compared with cargo-mission personnel who also flew in Southeast Asia but were not exposed to Agent Orange. The results reported are viewed as preliminary in that reproductive events were initially ascertained by self-report and are only now being verified in birth records; the results o f this validation process are expected shortly. The preliminary data showed a significant excess o f neonatal deaths (associated with severe defects) and o f physical handicaps among children o f Ranch Handers; overall, birth defects were 4nr 4oc 0010830 CA all hos cor For los: we: tha bin ju d Ra Th Th sta boi CC the bet sui ex: ese ju c the b> of re A: be w< tis at ex ST defense xnpare i f light 1 Delta s weee quaeies ` V_ ied the memed vtality. . ik the ciation . Cleft od and ; casedefects xl time ; were as Jon drome am be loped, teotial e that ops in iity in bicide jsures inated idose bicide 1 with * d to were srds: data ndof were Herbicide Exposure in Vietnam 249 more frequent in the Ranch Hand group, but the increase is statistically significant only when minor anomalies (eg, skin) are included. Other reproductive parameters considered (infertility, miscarriage) are not appreciably different in the two groups. CDC's A tlanta Study o f B irth Defects and Vietnam Service In August 1984 results o f the CDC birth defects study were published [5.6]. Cases, gathered from the Metropolitan Atlanta Congenital Defects Register, include all severe defects diagnosed through the first year o f life. Controls, matched for hospital, year o f birth, and race, were identified from birth certificates. Case and control parents were traced with similar procedures and interviewed by telephone. For mothers the overall response rate was 70% , for fathers only 56% (with most losses reflecting a failure to locate rather than a refusal). Among whites, the losses were equivalent in cases and controls while among nonwhites significantly more cases than controls were lost to interview. No association was found between father's reported Vietnam service and either birth defects in the aggregate or specific types o f malformations. Agent Orange exposure was also assessed using (1) self-reports and (2) an exposure index based on judgments by the military about where and when the man served in Vietnam (eg. Ranch Handers were generally assigned a score o f 5, representing highest exposure). These analyses are based on smaller samples than the analysis o f Vietnam service. The exposure index showed no association with birth defects overall but did show statistically significant associations with spina bifida, deft lip ( cleft palate), colobotna, and neoplasms in the first year o f life. C O N C L U S IO N S Our primary intention in this paper has been to provide detailed descriptions of the unpublished research on reproductive sequelae o f herbicide exposure that has been carried out by Vietnamese scientists. In order to put this in context we have also summarized the design and results o f the other reproductive studies known to us that examine populations potentially exposed in Vietnam (U .S .. Australian, and Vietnam ese soldiers, civilian residents o f Southern Vietnam). It remains for the reader to judge the quality o f each study and the compatibility o f the findings. In dosing, we venture three general observations and recommendations based thereon: 1) It can be argued that virtually all o f the research to date has been conducted by groups that are p a rti p ris to the Vietnam conflict (to a lesser extent this is also true o f the studies o f exposed occupational groups and Seveso residents). I f further research is undertaken, it might well be under the auspices o f an independent body. 2) Measurement o f exposure has thus far often been lacking or quite crude. Apparent inconsistencies in research results might be resolved if exposure dose were better quantified. Route o f absorption and environmental matrix (eg, soil, food) as weU~ as amount o f chemical should be considered. Biological markers (eg. adipose tissue levels) need to fte developed and applied in order to determine effective or absorbed dose following exposure. Data analysis should consider carefully specified exposure-effect models. 3) Timing o f exposure in relation to pregnancy has rarely been evaluated in the studies to date but needs to be taken into account. This is particularly true in the 4889 0010831 attest mod 250 Constable and Hatch studies o f paternally mediated birth defects, where a biologic model for teratogenesis (as distinct from mutagenesis) is lacking. REFERENCES 1. Friedman JM: Doe t Agent Orange cause birth defects? Teratology 29:193-221. 1984. 2. Hatch M : Reproductive effects of the dioxins. In Lownncc WW (ed) "Public Health Risks of the Dioxins." New York: The Rockefeller University, 1984, pp 255-274. 3. ta r a JH: Teratogens and the male. Med I Aim 2:16-20, 1983. 4. Young AL. Kang HK. Shepard BM: Chlorinated dioxins as herbicide contaminants. Environ Sri Tech 17:530A-540A, 1983. 3. Erickson JD. Mulinare J. McClain PW, Fitch TG . James LM . McClearn AB. Adams Jr. MJ: Vietnam veterans' risks for fathering babies with birth defects. JAMA 232:903-912. 1984. 6. Erickson JD. Mulinare J, McClain PW. Fitch TG. James LM . McClearn AB. Adams Jr. MJ: "Vietnam Veterans' Risks for Fathering Babies with Birth Defects." Atlanta: Cernera for Disease Control. 1984. 7. Lathrop CD. Wolfe W H. Albanese RA. Moynahan PM: "Project Ranch Hand II. An Epidemiologic Investigation of Health Effects in A ir Force Personnel Following Exposure to Herbicides: Baseline Morbidity Study Results." San Antonio. Texas: U.S. Air Force School of Aerospace Medicine. Aerospace Medical Division. Brooks A ir Force Base. 1984. 8. Nguyen Can, Nguyen Thi Xiem. Tran Tan Hong, Nguyen Kim Tong. Do Binh Duong: "An Epidemiological Survey of Pregnancies in Viet Nam." 1983a.* 9. Nguyen Can. Nguyen Thi Xiem. Nguyen Kim Tong, Do Binh Duong: "A Case-Control Survey of Congenital Defects in My Van District* Hai Hung Province." 1983b.* 10. Ton Duc Lang, Ton That Tung, Do Due Van: "Mutagenic Effects on the Fust Generation After Exposure to `Orange Agent'." 1983a.* 11. Ton Duc Lang, Do Due Van. Dwyer JH. Flameahaum C. Dwyer KM. Fantini D: "Seif-repons of Exposure to Herbicides and Health Problems: A Preliminary Analysis of Survey Data From the Families of 432 Veterans in Northern Viet Nam." 1983b.* 12. Nguyen Dinh Khoa: "Some Biologic Parameters Collected on the Groups of People in an Area Affected by Chemicals." 1983. 13. Ho Dang Nguyen: "Pregnancies the Polyclinic of Tty Ninh Province." 1983.* 14. Nguyen Thi Ngoc Pbuong, Le Thi Diem H in t: "The Effects ofToxic Chemicals on the Pregnancy of die Women Living at Two Localities in the South of Viet Nam." 1983.* 13. Meadson MS, Wearing AH. Constable JD: "Background Material Relevant Presentations Con cerning the HeAicide Assessment Commission for the Americas Association for the Advancement of Science." Washington: AAAS. 1971. Addendum to the Report. 16. Cutting RT. Phuoc TH . Balk JM , Detwrwon MW . Evans CH: "Congenial Malformations. Hydatidiform Moles and Stillbirths in the Republic of Vietnam, 1960-1969." Washington. D.C.: U.S. Government Printing Office, 1970. 17. Cling BindsTning. Nguyen Tran Chien: "Spontaneous Abortions and Birth Defects in Area Exposed to Toxic Chemical Sprays in Ghmg TromDistrict. Ben Tie Province. South Vietnam." 1983.* 18. Le Diem Huong. Nguyen Thi Ngoc Phuong: "The State of Abnormal Pregnancies and Congenital Malformations at the Gyneco-Obstetricml Hospital of Ho Chi MinhCity (formerly Tu Du Hospital)." 1983.* 19. Jacobs PA. WBsou C M , Sprenkle JA. Roacaahem NB, Migeon BR: Mechanism of origin of comptera hydaridifonn motet . Nature 287:714-716.1980. 20. Kuaatadrar P: "A Study o f Herbicides and Birth Defects in the Republic of Vietnam: An Analysts o f Hospital Records." National Academy of Sciences. Washington. D.C.: National Academy Press. 1982. 21. Donovan JW, Adetra M A . Rose G , Batistuta D: "Case-control Study of Congenital Anomalies and Viearam Service (Birth Defects Study)." Canberra: Australian Government Publishing Services. 1983. f Vietnamese papers presented at the Symposium on Herbicides and Defoliants in W ar The Long-term Consequence! on Man and Nature Ho Chi Minh City, January 13-20. 1983. 4890 'P 'H le C e 00J0832 I%( ' 4891 P -W 7 -J -> A/. 2 7 '-2^ n< /i nTECHNICAL PAPERS 0 < -fS '/ v -'-^ < - _ .'z.-c c-s^. y/-/-/ . * ./<_ Effects of 2,4-D on Seed Germination Seeds of barley and rice were first chosen as test materials. and Respiration These cereal grains, which are more or less alike in structure and food content, differ physiologically in that rice excels in -- ' fermentative faculty and is alone able to germinate in the O Y. L. Hsnssand C. H. Lou absence of oxygen (3 ). Seeds of uniform size and appearance O . Physiological Laboratory, were selected from the pure line stock kindly furnished us Tsing E v a U niversity, K ttnm ing, China by the Department of Plant Pathology, Tsing Hua University, ^ and then divided into lots of 100 seedseach. The different lots ^ _ Synthetic plant-growth-promoting substances, known also of seeds were treated with various dilutions of 2,4-D ranging msauxins, have been found to possess many practical applica from 0.0033 to 0.1 per cent. After 24 hours of treatment, each tions in agriculture. They have been extensively used to pre lot of seeds was thoroughly washed with water to remove the vent premature shedding of fruits, to induce rooting of adhering chemical and put in a labeled Petri dish in which a cuttings, to induce parthenocaxpy and ensure fruiting, to moist filter paper had been placed. The Petri dishes were CO keep the bud in dormancy, and to improve the well-being of assembled in a moist chamber mainnim-H at 25 2*G and --- a plant in general (4). The reasons for the versatility of auxin the rate of germination of the seeds followed at suitable inter treatment are still being conjectured. Only at very high dilu vals. As long as the filter paper in the dish is kept moist, addi tions (1 ppm) does indole acetic add promote growth and tion of excess water is avoided. In these experiments accelerate protoplasmic streaming in the A etna coleoptile; at lightly higher concentrations (10-100 ppm) it depresses growth, stunts the plant, but induces root formation; and at still higher (100-1,000 ppm) it becomes destructive to chloro phyll and to the plant as a whole (f). The recent successful introduction of di- or trichlorophenoxyacetic add, a compound \ which possesseshigh auxin activities, asa translocated herbiddal spray by F. D. Jones and others has amply revealed the importance of this new phase of research on auxin treatment. Furthermore, it has been shown that the herbiddal effect of this and allied chemicals on plants can be differential and selective, ix . it can be applied effectively in the field to loll some major weeds while leaving the crops unharmed (2). The facts that 2,4-D can be actively absorbed by. and F m .l. T h t (tra in *t n of barley ne*s lcctad by treatmentwith freely translocated in, the plant before the latter is killed, different eooctnuaiiou oi 2,4-D. and that its lethal effect on plants can be differential, seem to us most interesting and deserving of some physiological in the criterion for germination used by Taylor (3) was adopted, vestigation. m . seeds were regarded as germinated when any organ had Germination of seeds involves many of the physiological attained a length of not less than 1.5 mm The subsequent and biochemical processes exhibited in mature plants, such growth of the seedling was not included in our observations. as the breakdown and translocation of the stored food, the The 2,4-D employed in these experiments was synthesised formation of seedling organs, and the high respiratory and in our laboratory. activities during germination. If a chemical com The rates of germination of the treated seeds are plotted in pound h*ppn to be injurious to certain plants and if it hap Fig. 1. Similar to other auxins, 2,4 D accelerates seedgermina pens to be involved in inhibiting tome specific metabolic tion at low concentrations but delays it when a certain thresh activity therein, its physiological effect can be conveniently old of concentration is exceeded. The outstanding feature put to test by treating germinating seedswith sucha chemical, of our results is that the threshold concentration in retarding the dosage of which may be easily controlled. Various kinds of germination is higher in rice (ca. 0.07 per cent) than in barley seeds, differing in structure, in their principal food constitu (ca. 0.01 per cent). Furthermore, 0.07 per cent 2,4-D inhibits ents, and in tM 'r biochemical behavior, can be employed for barley germination completely while it merely delays germi the test. These differences may be expected to manifest them nation in rice. Even when the concentration is raised to 0.1 per selves through differences in their reaction toward a given cent, 2,4-D fails to prevent the rice from germinating, though chemical treatment This procedure was employed by us to it delays the process still further. Comparison of the germina study thephysiologicalaction of 2,4-D on plants. The chemical tion rate of seeds treated with 2,4-D with that of seeds kept treatment of seeds usually results in a complete inhibition of, under aerobic and anaerobic conditions reveals a striking re or a substantialdelay in, germination, which, on refined analy semblance between the treated seedsand thoseunderanaerobic sis, may reveal somespecific metabolic disturbance in the seeds. condition. It seems as if oxygen were no longer available to 0012200 2 P- I*?* 4892 mentation, for energy supply. Hence, during the few days following the chemical treatment, these seeds, in comparison with the control, exhibit a low oxygen uptake and a high carbon dioxide evolution, and, consequently, a high CO<:Ot ratio. This effect is very similar to that caused by germinating seedsunder low oxygen tension, which has been fully described by Taylor (J). As a result of 0 deficiency in the medium, aerobic respiration is reduced, and this reduction is to some extent compensated for by the fermentative activity of the germinating seeds, as evidenced by the high COj'.Oi ratios under such a condition. If they cannot furnish the energy necessary for germination fermentatively, most seeds will fail to germinate at alL If, as in rice, the seed is especially gifted with a highly functional fermentative mechanism, it can proceed to germinate, although with some delay, even after 2,4-D treatment or under anaerobic condition. However, this condition cannot go on indefinitely. Even rice cannot continue to grow in the complete absence of oxygen. During recent years evidence has accumulated which shows that auxin is involved in the 4-carbon add respiratory system (4 ). The increased respiration due to auxin treatment was found to parallel the increase in elongation of, and the acceleration of protoplasmic streaming in, the Avena coleoptQe. This statement is probably true only at certain low con centrations of auxin. Our experiments have shown that 2,4-D , at low concen trations (0.01 per cent), promotes germination; but, at higher concentrations (0.1 per cent), it begins to inhibit aerobic respiration and checks germination. R e fe re n ce s 1. Bozcm Ox, G. T ktM u h t h rau siM i /fUnU. 193*. P. M2.' 2. S u sa , JL E., Tu k x k a x , W. G,, sad Ssmow, W. A. N th trt, L n i., 1945, 159. 497. 3. TAUoa,D.L. Aaw ./ . * / ., 1943,29,731. 4. Vak Ovaaaxxx, J. Atom. X u. tirdum ., 1944, U . 631. Prevention o f R espiratory Em barrassm ent in Therapeutic Curarized Convulsions Matthxw B xody Departm ent o f N europsychiatry, B rooklyn ami Jew ish H ospitals, B rooklyn, New Y ork Transitory asphyxia and cyanosis occur so frequently in spontaneous and therapeutically induced convulsions that cerebral anoxia was cons dered a possible explanation for the beneficial effects of convulsions in mental disorders. However, it hasbeendemonstrated that the production ofcerebral anoxia by the inhalation ofnitrogen hasno suchtherapeutic value (4 ). On the contrary, asphyxia contributes clinically to strain and plays a role in the fatalities and near-fatalities that occasionally occur in convulsions. The treatment of asphyxia by the use of chemical stimulants has proved futile (J ). Ad ministration of oxygen is ineffectual unless the air passagesaxe open.The presenceof trismus makes the installation ofappara tus to clear the respiratory passagesafter a convulsion difficult (2) and often impossible. We havefound thatalthough admin istration of the convulsant electric current during inspiration facilitates postconvuisive breathing, since the first respiratory movement is then an expiration which dears the passages^! this so increases intrapulmonary and venous pressure as t P make the procedure not without risk. This procedure h a lp therefore, been discarded. Certain few patients are "hard breathers" in that they sistently have asphyxia! episodes during and after convulstwr"* seizures. They seem to fall into two main categories: (1) thosALTI whose illness is characterized by agitation, depression, rejecO ) tion of foodand sleep,and otherpsychological "oral" qualitiesrrs and (2) patients with evidence of generalized arteriosclerosiso ^ other neurologic complications, in whom there appears to b ,^ someinefficiency in the respiratory apparatus. The physician is sometimes compelled to treat a patient with coronary whose agitation is so marked and dangerous to the heart as to necessitate the administration of shock treatment in an at tempt to terminate the mental disorder. It is precisely those patients falling into both categories who are apt to show re spiratory difficulties and in whom asphyxia is most dangerous. In these patients it is impossible to soften the convulsion ade quately by means of curare because it increases respiratory difficulty and asphyxia. By premedication with sodium pentothal or sodium amytal (2) one can increase slightly the dosage of curare. The sedation seems to diminish the precanvulsive anxiety and restlessness and to diminish pharyngeal spasm. Unfortunately, these barbiturates increase the postconvulsive apnea. We have found a simple mesns of avoiding asphyxia and consequently diminishing cardiac strain. With this technique a Guedel robber airway is installed during the convulsion. The patient is prepared as usual for the treatment. He is placed supine on a flat bed without hyperextension, since the latter increases respiratory difficulty. The barbiturate and curare, or curare tim e, are injected. In electroshock convul sions, a cloth gag is placed between the teeth to protect the tongue and lips. The current is then applied. During the initial tonic flexion phase, the gag is forcibly bitten. This is followed by a moment of relaxation in which the mouth is opened widely. At this point a Guedel rubber airway is introduced to the hilt. Where metzazolis used, thecloth gag isnot necessary. The initial movement of the mouth is an opening one, and the airway is similarly introduced. With the airway in place there may be respiratory exchange in the midst of the convulsion. Before the end of the convulsion, the patient is turned on the side with the mouth down. Mucus and saliva will flow from theairway. Respiration usually beginsshortlyafter the convul sion' is over. The airway is removed a few minutes after the convulsion, when respirations are normal and the mouth and jaw relaxed. With this technique,asphyxia and cyanosis are now rare. There are pension! periods of apnea, which seem to be of central origin. In these apndc periods it is a simple matter, with the air passagesopen, to instituterespiration, if one so de sires, by abdominal pressure. The airway acts as an efficient mouth gag and bitten tongues and lips are infrequent. A theoretical objection to the method is the possibilityof disloca tion of the jaw and trauma to the pharynx, but in practice neither occurs. Postconvulsive headache, nausea, and confu sion have been diminirfwH considerably. Larger amounts of curare can be administered safely to hard breathers. In one patient with known severe coronary disease, the postconvul siveelectrocardiographicchangeswereleu marked andof much 0012202 4893 S699ii?Mnni King, Joseph E. and Penfound, William T. Effects of Two of the New Pormkgenic Herbicides on Bream and Largemouth Bass. E cology 27, No. 4, 372-374 (1946); through Biological Abstracts 21, 19017T1947). --- Tests were made on 2,4D to determine its effect on fingerllng bream and largemouth bas3. The fish were subjected to the following conditions: tapwacer control, 1 p.p.m. of herbicide, 100 p.p.m. of herbicide, under hyacinth plants as a control, and under hyacinth plants sprayed with 1,000 p.p.m. of herbicide. Solutions of 1 p.p.m. of 2,4-D had no effect; whereas 100 p.p.m. produced a low but significant mortality. Pish under hyacinth plants sprayed with 1,000 p.p.m. 2,4-D experienced no ill effects until the plants began to die and the subsequent oxidation lowered the dissolved 0g content of the water beyond the toleration limits of the fish. The results indicate that, whereas the introduction of 2,4-D into the immediate environment of the fish may have harmful effects, spray applications to surface vegetation at the rates recommended by the manufacturer will not be directly toxic to fish life. When the sprayed plants die, however, any fish trapped in the immediate vicinity and unable to escape to open water will probably die of asphyxiation. 0012204 V - m o 4894