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IN THE UNITED STATES DISTRICT COURT FOR THE EASTERN DISTRICT OF TEXAS BEAUMONT DIVISION 1 CECIL SCOTT, ET AL v. MONSANTO COMPANY ] ] No. B-84-1103-CA ] EXHIBITS TO VIDEOTAPE DEPOSITION OF DR. GEORGE LEVINSKAS May 28, 1987 1300 Post Oak Boulevard Houston, Texas Wanda G. Kuhn, Court Reporter Nell McCallum a Associates Inc. 2900 Smith, Suite 104 Houston, Texas 77006 (713) 523-3767 NELL MC CALLUM & ASSOCIATES, INC. WATER PCB-SD0000012540 m N T-t C/i r<:r o i-t 5--5i XeCinau--cut3aoa-/.i <tnuoaJ. CO <fl Q. OS U0>5J CH nOJ XI-I > CJ 4(JBOBl "> U tot. Ot 040 >H>oo OV U nH CJ ix--I a aS5 o .. IS <P COM CoMI QMc.rt i A'*' s DM EH C'*: Toxicology Section/St. Louis tcojotvjoerr^LOCATioN) SPECIAL (TYPE OF REPORT) REPORT REPORT NO.: MSL-2002 JOB/PROJECT NO.: DATE: October 14, 1981 TITLE: TOXICITY OF AROCLOR PRODUCTS 1242,; 1254 AND 12 60 TO THE LIVER OF ALBINO-RATS AUTHORS: George J. Levinskas, Ph.D. ABSTRACT: This report is a tabulation of the results of microscopic evaluation of liver sections from rats fed AROCLOR 1242, AROCLOR 1254 or AROCLOR 1260 for two years. n-iaa(a> SCM 058930 000007 WATER PCB-SD0000012541 DISTRIBUTION COPY NUMBER 1 - Reports.Library, R2C 2 - Reports Library, R2C 3 - Reports Library, R2C 4 - DMEH Library, G2WA 5 - T3MEH Library, G2WA 6 - R.T. Berendt, E2ND 7 - J.H. Craddock, A2SA 8 - G.J. Leviriskas, G2WF 9 - J.G. Nassi'f, E2ND '' 10 - J.M. Norris, Dow Chemical 11 - R.A. Stohr, B3NJ ABSTRACT ONLY This report has been assigned to you. When it is no longer needed, you are responsible for returning it to: ___________ REPORTS LIBRARY, R2C __________ If you transfer it to anyone else, please let your librarian know, so the records can be changed. R>10II(C) (RCV. l/M) OOOCG'^cm 05893 L WATER PCB-SD0000012542 INTRODUCTION The polychlorinated biphenyls (PCBs) comprise a family of compounds of variable chlorine content. PCBs manufactured by Monsanto (tradenamed "Aroclor") bear a four digit number, the '12' indicating biphenyl and the last two digits indicating the chlorine content by weight percent. Since the chlorine atoms are randomly distributed among the 10 ring positions available for substitution, each material is a mixture of isomers. In 1969, Monsanto sponsored a series of animal studies at Industrial BIO-TEST Laboratories in Northbrook, Illinois, on Aroclor 1242, 1254, and 1260 for the assessment of the health and environmental hazards of these materials.. This series consisted of 2-year chronic feeding studies to rats and dogs, a 3-generation rat reproduction study, a rat teratology study, a dominant lethal mutagenic study in mice and a toxicity/reproduction study in chickens on each of the 3 Aroclor products. Even though no such action was contemplated, such a broad battery of tests would have been adequate to support Food Additive Petitions for each of these materials. These studies were initiated by reports that PCBs had been detected in the environment. A report (Nelson, 1972b) by a Panel on Hazardous Trace Substances of an Ad Hoc Committee on Environmental Health Research covers the development of information on the environmental and biological effects of PCBs. There have been subsequent literature reviews which need not be recapitulated here [DHEW, 1978; EPA, 1976; IARC, 1978; NAS, 1979; Roberts, et al. (1978)]. Starting in 1969, while these studies still were in progress, information regarding them was made available to various government groups.1 Subsequently, data from these studies were presented at a conference '. - i- 000CG3 SCM 058932 WATER PCB-SD0000012543 sponsored by the National Institute of Environmental Health Sciences at Rougemont, N.C. on December 20-21, 1971, but the account of these studies was omitted from the published proceedings (Keplinger, et al., 1972; Nelson, 1972a). Subsequently, it was reported that female Sherman strain rats developed hepatocellular carcinomas when fed Aroclor 12602 from Lot No. AK-3; the same lot used for the earlier Monsanto study with this material. As a result, Monsanto requested the contract laboratory's pathologists to review livers from all available rats from the 3 Aroclor studies. That review (which could have included new sections of liver from animals examined previously in addition to sections from animals not examined previously) was presented in the reports of Gordon and Richter (1975a,b,c). In November 1975, reports containing evaluations of liver sections from the 2-year rat feeding studies (Gordon and Richter, 1975a,b,c) and the results of an independent evaluation of the same liver sections (Pour, 1975) were presented to and discussed with several federal regulatory agencies3. Later that month, a summary of data from all of the studies was presented at the National Conference on Polychlorinated Biphenyls sponsored by the Environmental Protection Agency and held in Chicago on November 19-21, 1975 (Calandra, 1976). ' Only summaries of data from these and other toxicity studies conducted over the years have been published (Jenkins, et al.. 1972; Keplinger, et al., 1971; Monsanto, 1980). Knowledge of these efforts may have prompted the statement in a recent article that "...Monsanto, whose reaction to the possibility that polychlorinated biphenyls (PCBs) might be an ecological disaster was a classic of how such issues should be handled, says Ford4. Monsanto began to investigate the dangers as soon as they were 2 - ooocoo sc* 059,33 WATER PCB-SD0000012544 seriously voiced. It insisted on keeping an open mind about them. As soon as evidence appeared that there was a strong chance PCBs were a hazard, it published the results of its investigations, admitting the danger. It thus established a reputation of honesty even among the environmentalists.'1 (Clutterbuck 1981). At a later meeting in Bethesda, MD.S, pathologists selected and reviewed some liver slides from the Monsanto-sponsored and Kimbrough studies. The pathologists differed in the terminology used to classify the lesions. They did conclude that the general incidence and severity of liver lesions (hyperplasia, nodular hyperplasia and neoplastic changes - carcinomas) were greater in the rats from the study subsequently published by Kimbrough, et al. (1975). No carcinomas were seen in the Monsanto-sponsored studies. It was noted that either the strain of rat or sex could have accounted for the differences. The spontaneous incidence of hyperplastic or neoplastic liver lesions in the Sherman strain rat was unknown, and the occurrence of such lesions appears to be higher in female rats and mice. The different diagnoses were discussed with Dr. Philippe Shubik of the Eppley Institute for Research in Cancer at the University of Nebraska. Dr. Shubik suggested that the liver sections from these studies could be reviewed by Dr. Parvis Pour. This was done. This publication is an effort to make readily available information in the Gordon and Richter reports (1975a,b,c) which are only summarized in the literature (Calandra, 1976). 000-01 - 3 SCH 058934 WATER PCB-SD0000012545 METHODS It appears that the Threshold Limit Value of 0.5 mg/m3 for chlorobiphenyl with 54% chlorine (ACGIH, 1969) was used to estimate suitable dose levels for the rat feeding studies. For that purpose the following assumptions were made: if the ambient air concentration of PCBs is 0.5 mg/m3, and if all of the inhaled PCBs are absorbed, and if 15 m3 of air are inhaled during the working day, a person would receive a PCB dose of 7.5 mg/day. The corresponding dosage would be approximately 0.1 mg/kg/day for a 70-kilogram person. Consequently, dosages of 0.1, 1, and 10 mg/kg/day were decided upon, and the dietary concentrations were set at 1, 10, and 100 ppm. As it turned out, the assumptions used to set dosages for the feeding study resulted in the low dose level rats receiving a dosage that was 100 times higher than the 0.001 mg/kg/day which FDA later calculated as allowable for protracted ingestion based on human data (FDA, 1973). For the chronic toxicity study in rats6, weanling animals of the Charles River CD strain7 were divided into a control group and 9 treatment groups, each consisting of 50 males and 50 females, with 3 treatment groups assigned to each of the 3 Aroclor products studied (1242, 1254, and 1260). Not all of these animals started at the same time. After about 2 months on test, additional groups of males and females were assigned to each test diet and the controls. These animals were added to allow a sufficient number of animals for sacrifices at 3, 6 and 12 months to provide some information prior to the completion of the 2-year study period. The numbering sequence (noraz 4- SCM 058935 WATER PCB-SD0000012546 and the designation of some animals as "Extra" suggests that additional animals were placed on test. Groups of 5 males and 5 females were scheduled to be sacrificed after 3, 6, and 12 months of feeding, and survivors were to be sacrificed at the end of the test period.* Animals were to be given a gross autopsy and tissues were to be preserved for possible histologic examination. Tumors or lesions suggestive of tumors were to be examined. RESULTS Data from the Gordon and Richter reports (1975a,b,c) on liver slides are shown in Tables 1, 2, and 3 for Aroclor 1242, 1254, and 1260, respectively. While the text of the reports contained comments specific to the particular Aroclor, they also contained similar comments such as "There is evidence of a chemical effect on the liver......... This consists of a hepatocellular alteration beginning as focal hypertrophy which progresses to nodular hyperplasia, and in a few animals to hepatoma or cholangiohepatoma" and "In the absence of metastasis, invasiveness, severe basophilia, mitoses, or other evidence of anaplasia; these are benign tumors rather than malignant carcinomas. There was no evidence of metastasis or invasiveness of these tumors in this study". The reports also stated that "The other treatment-related lesions reported are regarded as degenerative or hyperplastic in nature and they are morphologic manifestations of an adaptive response of the liver associated with biotrans formation of the test material" and "In most instances, the spectrum of treatment-related histopathological findings in the liver from this re-evaluation did not differ significantly from that previously reported in our original report......... No hepatocellular carcinomas were observed". 5- OSCHV,05:S936 WATER PCB-SD0000012547 With respect to Aroclor 1254, it was noted that "One liver tumor (a hepatoma) previously reported in a T-II animal (No. 445) was re-classified as nodular hyperplasia" (Gordon and Richter, 1975a)1. DISCUSSION The initial reports of these studies concluded that the target organ was the liver for each Aroclor product. This was confirmed by the second evaluation of liver slides (Gordon and Richter, 1975a,b,c). These alterations were slow to develop, their incidence being related to both dose level and duration of treatment. Since there were increased incidences of vacuolar changes in the cytoplasm of the hepatocytes and focal hypertrophy at all dose levels for all 3 Aroclor products at the 24-month sacrifice, a "no-effect" level was not established for liver effects. With respect to the slides from Industrial BIO-TEST, Pour (1975) concluded that Aroclor 1242, 1254, and 1260 "showed a dose-dependent hepatotoxic effect, characterized by degenerative and regenerative processes. With one exception, all lesions were considered non-neoplastic. Structures similar to cholangiocarcinomas and hepatomas were found in one rat. However, the possibility of metastases of a carcinoma into the liver has been also considered." In the report, he noted one lesion which seemed to "represent a metastatic tumor (probably a mammary gland carcinoma of the same animal from which ...(the particular liver section]... was submitted), rather than a cholangiocarcinoma". This occurred in an animal fed 100 ppm of Aroclor 1254. The contract laboratory pathologists diagnosed the presence of mammary tumor metastases in the liver of this rat (Gordon and Richter, 1975b).11 -s- - SCM 058931 WATER PCB-SD0000012548 SUMMARY and CONCLUSIONS Groups of male and female rats were fed diets containing either 1, 10, or 100 ppm of one of 3 Aroclor products, 1242, 1254, or 1260, for 2 years. Focal hypertrophy of the liver occurred at 3, 3, and 12 months for rats fed Aroclor 1260, 1254, and 1242, respectively. Focal hypertrophy was not seen in livers of control animals. At the 24-month sacrifice, livers of animals from all dose levels of the 3 Aroclor products had an increased incidence of vacuolar changes and focal hypertrophy. Livers of some animals fed 100 ppm of each Aroclor also had benign liver tumors (hepatoma or cholangiohepatoma). No hepatocellular carcinomas were observed. 7- SCM 058938 WATER PCB-SD0000012549 Footnotes Dates of initial correspondence and contacts. October 3, 1969. E.P. Wheeler to A.R. Glasgow, Division of Pesticides. Food and Drug Administration. April 8, 1970. R.E. Kelly to H. Blumenthal, Petitions Review Branch, Food and Drug Administration. ' April 22, 1970. E.P. Wheeler to E.J. Burger, Jr., Office of Science and Technology. June 1, 1970. E.P. Wheeler to H.E. Stokinger, Bureau of Occupational Safety and Health. R.D. Kimbrough, personal communication. November 13-14, 1975. Council on Environmental Quality. Environmental Protection Agency. Food and Drug Administration. House Subcommittee Manpower, Compensation, Health and Safety. National Cancer Institute. National Institute for Environmental Health Sciences. National Institute for Occupational Safety and Health. Dr. David Ford of Bath University. At National Cancer Institute on January 21, 1975. Present were D.E. Gordon, R.D. Kimbrough, G.J. Levinskas, R.A. Squire, W.R. Richter. _ . Since the events described earlier, the validity of many toxicity studies conducted by Industrial BIO-TEST Laboratories has been challenged. Therefore, the available raw data supplied by Industrial BIO-TEST was reviewed to determine whether this study could be validated. The review showed that the data bases (including the lack of a protocol) were insufficient for a complete validation of the study. It was decided to focus on presenting the primary liver effects reported by Gordon and Richter (1975a,b,c). Therefore, a complete audit was not undertaken. Available records were examined for a determination that the animals were placed on test and of their ultimate fate. Necropsy and microscopic reports were also examined for findings pertaining to livers of those animals. Significant discrepancies which were found between data in Tables 1, 2, and 3 and the data base for the Gordon and Richter reports are noted in this report. On some records, the labels Aroclor 1242 and Aroclor 1260 are interchanged as determined by a check of the animal numbers. Charles River Breeding Laboratories, North Wilmington, Massachusetts. Animals sacrificed at 6 and 12 months were placed on test about 2 months after the first group. Those sacrifice periods are sometimes labeled 8 and 14 months, respectively, which corresponds to the calendar time from the start of the test but overstates the time on test for those groups. OnQ-36 - 8 058^ sort WATER PCB-SD0000012550 9. As a result of the examination described in footnote 6, a few animals were reassigned to other dose levels on basis of assigned numbers. Three with no recorded liver lesions were deleted from the 24-month listing because of short duration on test: 14 months at 100 ppm Aroclor 1242, 15 months at 10 ppm Aroclor 1254 and 13 months at 100 ppm Aroclor 1254. Therefore, numbers in Tables vary slightly from those in reports of Gordon and Richter (1975a,b,c). 10. That change and comments in the footnotes to the Tables were noted ' during the examination described in footnote 6. In addition, the following also were noted during the examination. Aroclor 1254 - 100 ppm animal marked "Extra3" with diagnosis of hepatoma in record of examination for original Industrial BIO-TEST report. Animal not listed in Gordon and Richter report. . - 100 ppm animal no. 542 with gross autopsy notation that liver appears tumorous and white foci has no record of examination. Aroclor 1260 - 100 ppm animal no. 293 with gross autopsy notation of . tumor on liver has no record of examination. In some instances, diagnoses were crossed out on the available records. These were included in the Tables. Crossed out diagnoses for hepatoma or cholangiohepatoma are noted in the footnotes to the Tables. 11. Dr. Pour's report does not include the following liver sections noted by discrepancies between his tabulation and the Gordon and Richter reports. 1 from control at 12 month sacrifice, 5 from 100 ppm Aroclor 1242 at 24 months, 1 from 100 ppm Aroclor 1260 at 3 months. None of these animals were reported as having hepatomas or cholangiohepatomas in the Gordon and Richter reports. 000C07 SCH 058940 WATER PCB-SD0000012551 References 1. ACGIH (1969). Threshold Limit Values of Airborne Contaminants. American Conference of Governmental Industrial Hygienists. Cincinnati. 2. Calandra, J.C. (1976). Summary of toxicological studies on commercial PCB's. In: National Conference on Polychlorinated Biphenyls, November 19-21, 1975. Chicago, Illinois. EPA Report No. 560/6-75-004. March. - NTIS PB-253 248. pp.35-42. 3. Clutterback, D. (1981). What causes environmental conflict? New Scientist 90, 176-177. 4. DHEW (1978). Final Report of the Subcommittee on Health Effects of PCBs and PBBs. Environ. Health Perspect., 24, 129-198. 5. EPA (1976). National Conference on Polychlorinated Biphenyls, November 19-21, 1975. Chicago, Illinois. U.S. Environmental Protection Agency. Washington, D.C. EPA-560/6-75-004, March. NTIS PB-253 248. 6. FDA (1973). Polychlorinated biphenyls (PCB's). Contamination of animal feeds, foods and food-packaging materials. Fed. Regis., July 6, 38, 18096-18103. 7. Gordon, D.E. and Richter, W.R. (1975a). Two-year chronic and toxicity study with Aroclor 1242 in albino rats. Histopathological evaluation of additional liver sections. March 24 (unpublished observations). 8. Gordon, D.E. and Richter, W.R. (1975b). Two-year chronic oral toxicity study with Aroclor 1254 in albino rats. Histopathological evaluation of additional liver sections. March 24 (unpublished observations). 9. Gordon, D.E. and Richter, W.R. (1975c). Two-year chronic oral toxicity study with Aroclor 1260 in albino rats. Histopathological evaluation of additional liver sections. March 24 (unpublished observations). 10. IARC (1978). Polychlorinated biphenyls. IARC Monographs on the evaluation of the carcinogenic risk of chemicals to humans. 18, 43-103. International Agency for Research on Cancer. Lyon, France. October. 11. Jenkins, D.H., Keplinger, M.L., Fancher, O.E., Wheeler, E.P. and Calandra, J.C. (1972). Reproductive effects of polychlorinated biphenyls in white leghorn chickens. Toxicol. Appl. Pharmacol. 22, 316. 12. Keplinger, M.L., Fancher, O.E., Calandra, J.C. (1971). Toxicologic studies with polychlorinated biphenyls. Toxicol. Appl. Pharmacol. 19, 402-403. 00QC33 SCM 0589*1 WATER PCB-SD0000012552 13. Keplinger, M.L., Fancher, O.E., Calandra, J.C., et al. (1972). Toxicological studies with polychlorinated biphenyls. Read at PCB Conference, Quail Roost Conference Center, Rougemont, North Carolina, 20-21 December 1971. Cited in Polychlorinated Biphenyls Environmental Impact. A Review by the Panel on Hazardous Trace Substances. March 1972. Environ. Res. 5 , 249-362. 14. Kimbrough, R.D., Squire, R.A., Linder, R.E., Strandberg, J.D., ' Montali, R.J., and Burse, V.W. (1975). Induction of liver tumors in Sherman Strain female rats by polychlorinated biphenyl Aroclor 1260. J. Natl. Cancer Inst. 55, 1453-1459. 15. Monsanto Company (1980). Polychlorinated biphenyls. PCB's. A report on Uses, Environmental and Health Effects and Disposal. 16. NAS (1979). Polychlorinated biphenyls. National Academy of Sciences. Washington, D.C. 17. Nelson, N. (1972a). Comments on research needs. Environ. Health Perspect., 1, 181-185. 18. Nelson, N. (1972b). Chairman, Panel on Hazardous Trace Substances. Polychlorinated biphenyls - environmental impact. Environ. Res. 5, 249-362. 19. Pour, P. (1975). Histopathological reevaluation of livers for rats treated with Aroclor. August 1 (unpublished observations). 20. Roberts, J.R., Rodgers, D.W., Bailey, J.A., Rorke, M.A. (1978). Polychlorinated Biphenyls: Biological criteria for an assessment of their effects on environmental quality. National Research Council of Canada. Ottawa. Publication No. NRCC 16077. SCH 058942 WATER PCB-SD0000012553 Sacrifice Interval Diet Level (ppm) Table 1 Aroclor 1242: Primary Liver Lesions Among Albino Rats 3 Months 0 1 10 100 6 Months 0 1 10 100 12 Months 0 1 10 100 a Terminal 0 1 10 100 No. Animals Examined Findings Vacuolar change Focal necrosis Focal lymphoid infiltration Focal hypertrophy hepatocytes Nodular hyperplasia Ductular hyperplasia . Hepatoma Cho1angiohepa toma 10 8 10 9 202 010 000 000 000 000 000 000 1 0 0 0 0 1 0 0 10 10 8 9 1 00 1 10 030 000 000 000 000 000 0 0 0 0 0 0 0 0 10 10 10 9 1 24 000 00 1 000 000 101 000 000 0 0 0 1 0 0 0 0 23 32 29 19 178 111 100 023 111 533 000 000 9 0 0 8 8 3 3b 1C * Control group has animal dead at 19 months and 2 marked "Extra". 1 ppm group haa animals dead at 19, 22, 22, 23, 23 months. 10 ppm group has animals dead at 23, 23 months. ,t100 ppm group has animals dead at 22, 22, 22 months and 2 marked "Extra". ^ Includes 1 animal without record of examination in original IBT report. Not listed as hepatoma in worksheets for ^ Gordon and Richter report but appears and was crossed out on typed tabulation for animal marked "Extra". O for other 2 animals do not appear in records of examinations for original IBT report. O'- . t-)C Diagnosis does not appear in records of examinations for original IBT report. Diagnoses SCM 0 5 8 9 4 3 WATER PCB-SD0000012554 Sacrifice Interval Diet Level (ppm) Table 2 Aroclor 1254: Primary Liver Lesions Among Albino Rats 3 Months 0 1 10 100 6 Months 0 1 10 100 12 Months 0 1 10 100 Terminal a 0 1 10 100 No. Animals Examined Findings Vacuolar change Focal necrosis Focal lymphoid infiltration Focal hypertrophy hepatocytes Nodular hyperplasia Ductular hyperplasia . Hepatoma Cholangiohepatoma 10 10 10 10 233 01 1 000 000 000 000 000 000 1 0 1 1 0 0 0 0 10 10 10 10 1 00 1 20 000 000 000 000 000 000 0 2 2 4 0 0 0 0 10 10 10 10 1 14 000 002 004 0 00 1 11 000 000 1 1 1 2 0 0 0 0 23 30 26 26 1 7 9 13 131 1 120 1 0 3 4 14 1 0 3 14 563 00 0 4 4b 000 2C * Control group has animal dead at 19 months and 2 marked "Extra". 1 ppm group has animals dead at 22, 22 months, 1 marked "Extra" and 1 other for which date of death is not recorded. 10 ppm group has animals dead at 22, 22, 22 months. ,,100 ppm group has animals dead at 23, 23 months and 9 marked "Extra". b Includes 2 animals marked "Extra". Diagnoses do not appear in records of examination for original IBT reports. o C3c Both animals marked "Extra" with same designation. 1 without apparent record of examination for original IBT report Q Diagnosis for other animal does not appear in records of examinations for original IBT report. b ' '1 H- WATER PCB-SD0000012555 Table 3 Aroclor 1260: Primary Liver Lesions Among Albino Rats Sacrifice Interval Diet Level (ppm) 3 Months 0 1 10 100 6 Months 0 1 10 100 12 Months 0 1 10 100 Terminal a 0 1 10 100 No. Animals Examined 10 10 iob 10 10 6 5 9 10 9 10 9 23 26 25 25 Findings Vacuolar change 202 3 11 Focal necrosis 0 0 4C 0 10 .Focal lymphoid infiltration 000 0 02 Focal hypertrophy hepatocytes 000 2 00 Nodular hyperplasia 000 0 00 Ductular hyperplasia 00 1 0 00 Hepatoma 000 0 00 Cholangiohepatoma 000 0 00 *. Control group has animal dead at 19 months and 2 marked "Extra". 2 ld 2 0 0 0 0 0 6 0 0 3 0 0 0 0 1 25 000 0 00 000 0 00 1 10 0 00 000 3 0 0 4 1 2 0 0 157 9 143 6 101 0 0 3 13 9 107 6 5 6 7 12 0 0 le 7f.g 000 48. * 1 ppm group has animals dead at 20, 22, 22 months. 10 ppm group has animals dead at 19, 22, 22 months and 1 marked "Extra". 100 ppm group has animals dead at 22, 22 months. ** Includes 1 marked "Extra". c Worksheets show listings under this diagnosis with arrow pointing to Vacuolar change colusw. d,Date of death of this animal not recorded. e No record of examination for original IBT report. Listed on worksheets for Gordon and Richter report with diagnosis crossed out. * Includes 2 animals without record of examination for original IBT report. Diagnoses for other 5 animals do not appear in 0 records of examinations for original IBT report. ^ reports. 2 of these diagnoses crossed out on worksheets for Gordon and Richter O Includes one animal with both diagnoses. Hepatoma is 1 of 2 crossed out (superscript f). Qh Includes 3 animals without record of examinationfor original IBT report. Diagnosis for other animal does not appear in tO records of examinations for original IBT report. 2 of these diagnoses crossed out on worksheets for Cordon and Richter reports. ^ to b ^ u MJb WATER PCB-SD0000012556 r WATER PCB-SD0000012557 IN THE UNITED STATES DISTRICT COURT FOR THE EASTERN DISTRICT OF TEXAS BEAUMONT DIVISION CECIL SCOTT. ET AL VS. MONSANTO COMPANY CIVIL ACTION NO B-84-1103-CA PLAINTIFFS' NOTICE OF INTENTION TO TAKE ORAL DEPOSITION AND SUBPOENA DUCES TECUM TO: Defendant, MONSANTO COMPANY, by and through its attorneys of record Mr. Robert A. Hall, Mr. Jonathan B. Shoebotham, Woodard, Hall & Primm, 4700 Texas Commerce Tower, Houston, Texas 77002; Mr. Walter J. Crawford, Jr., Wells, Peyton, Beard, Greenberg, Hunt & Crawford, P. 0. Box 3709, Beaumont, Texas 77704; Mr. John M. Johnson, Mr. Warren B. Lightfoot, Bradley, Arant, Rose & White, 1400 Park Place Tower, . Birmingham, Alabama 35203; and Mr. Michael R. Fruehwald, Mr. Michael Rosiello, and Ms. Anne C. McGown, Barnes & Thornburg, 1313 Merchants Bank Building, 11 South Meridian Street, Indianapolis, Indiana 46204. Pursuant to the provisions of Rule 30(b)(6) of the Federal Rules of Civil Procedure, the Plaintiffs in this cause hereby give NOTICE to the Defendant that the oral deposition of MONSANTO COMPANY, Defendant, will be taken on the date and time and at the place hereinafter stated. Specifically, the Plaintiffs intend to take the oral deposition of the representative(s) designated by the 2 DEPOSITION f EXHIBIT WATER PCB-SD0000012558 Defendant pursuant to Rule 30(b) (6) of the Federal Rules of Civil Procedure. Plaintiffs request that Defendant designate that current employee of Defendant having held the highest position of any held by a current employee in Monsanto Company's toxicology department during the period 1965 through 1975. Plaintiffs also request that Defendant designate that representative knowledgeable of the documents and subject matter herein below enumerated. 1) Any document (s) which would state or disclose the following: a) when Monsanto first began using the services of IBT; b) when Monsanto first began using the services of IBT in connection with Monsanto's PCB products; c) the dollars spent by Monsanto with IBT (i) each year that Monsanto used IBT's services, (ii) on all PCB related studies, and (iii) totally, from the date Monsanto first began using the services of IBT. 2) All correspondence from IBT to Dr. Paul L. Wright while Dr. Wright was an employee of Monsanto; 3) All correspondence by Monsanto to Paul L. Wright or from Paul Wright to Monsanto; 4) Any document stating, reflecting or referring to the policies and procedures of Monsanto's Toxicology Department for the years 1965 through 19 80; 5) Any document stating, reflecting or referring to the current standards, procedures and policies of Monsanto's Toxicology Department; -2- OOQCOZ WATER PCB-SD0000012559 6) The results of any IBT PCB test redone by or for Monsanto Company; 7) Any investigation or background file on Dr. Renata Kimbrough; 8) Any correspondence to or from Dr. William Ribelin; 9) Any report, analysis or investigation of IBT misconduct (actual or alleged) in connection with tests performed on Monsanto products (this request includes copies of government reports; independent investigations; internal IBT investigations; Monsanto authored or sponsored investigations; and, investigations by industry groups, bodies or authorities or other private or public body) (IBT includes all of its employees and consultants). 10) Any document which would in any way reflect, relate or pertain to a change by or on behalf of Monsanto to the results (interim or final) of any test of a Monsanto product by IBT; 11) Any document which would in any way reflect, relate or pertain to a change by or on behalf of Monsanto to the results (interim or final) of any test of a Monsanto PCB product by IBT; 12) Any document which would in any way reflect, relate or pertain to a suggestion by Monsanto regarding the outcome of a test to be conducted by IBT for Monsanto on any Monsanto product; 13) Any document which would in any way reflect, relate or pertain to a suggestion by Monsanto regarding the outcome of a test to be conducted by IBT for Monsanto on any Monsanto PCB product. This deposition will be taken at the offices of Gilpin, Pohl & Bennett, 1300 Post Oak Boulevard, Allied Bank Tower, G00C03 -3- WATER PCB-SD0000012560 23rd Floor, Houston, Texas 77056 beginning at 9:00 a.m. on Saturday, May 23, 1987 and shall continue until completed. This oral deposition will be taken before a certified court reporter. The testimony given during this oral deposition will be used as evidence in this matter, together with the documents produced at this deposition. You are invited to attend and dross-examine. As used in this notice, the term "documents" means any printed, typewritten or handwritten matter, or reproduction thereof, of whatever character, in the possession, custody or control of a witness or his agents, representatives or employees, including without limitation, correspondence, contracts, memoranda, agreements, letters, brochures, reports, handwritten or typewritten notes, sound recordings or transcriptions thereof, computer print-outs, inter-company and intra-company communications, work papers, diaries, calendar pads, appointment books, ledgers, financial statements, checks, bank drafts and writings of whatever kind and character, whether originals or reproductions, and whether in draft or final form, including copies bearing different markings or notations. "PCB" means polychlorinated biphenyl and its contaminants and derivatives. -4- ooooo** WATER PCB-SD0000012561 Respectfully submitted By:_______________________ Michael A. Pohl OF COUNSEL: David M. Lacey GILPIN, POHL & BENNETT 1300 Post Oak Boulevard Allied Bank Tower, 23rd Floor Houston, Texas 77056 (713) 623-8800 REAUD, MORGAN & QUINN 909 Laurel Avenue Beaumont, Texas 77701 (409) 838-9941 Thomas Henderson HENDERSON & GOLDBERG 1030 Fifth Avenue Pittsburgh, Pennsylvania (412) 471-3980 15219 Benton Musslewhite LAW OFFICES OF BENTON MUSSLEWHITE 609 Fannin, Suite 517 Houston, Texas 77002 (713) 222-2288 David S. McCrea McCREA & McCREA 119 S. Walnut Street Bloomington, Indiana (812) 336-4840 . 47402 ATTORNEYS FOR PLAINTIFFS, CECIL SCOTT, ET AL. CERTIFICATE OF SERVICE -5- QQQCOn WATER PCB-SD0000012562 I hereby certify that on the ______ day of , 1987 , a true and correct copy of the above Notice of Intent to Take Oral Deposition and Subpoena Duces Tecum was served upon counsel of record either by messenger or by placing same in the United States mail, certified mail, return receipt requested, postage prepaid and addressed as follows: Mr. Robert A. Hall Mr. Jonathan B. Shoebotham Woodard, Hall & Primm 4700 Texas Commerce Tower Houston, Texas 77002 Mr. Walter J. Crawford, Jr. Wells, Peyton, Beard, Greenberg, Hunt & Crawford, P. 0. Box 3708 Beaumont, Texas 77704 Mr. John M. Johnson Mr. Warren B. Lightfoot Bradley, Arant, Rose & White 1400 Park Place Tower Birmingham, Alabama 35203 Mr. Michael R. Fruehwald Mr. Michael Rosiello Ms. Anne C. McGown Barnes & Thornburg 1313 Merchants Bank Building 11 South Meridian Street Indianapolis, Indiana 46204 depntc02/txt8 65 51 Michael A. Pohl -6- OOQCC WATER PCB-SD0000012563 LUXI V \ I B I T i ji ii ! WATER PCB-SD0000012564 IN THE UNITED STATES DISTRICT COURT FOR THE EASTERN DISTRICT OF TEXAS BEAUMONT DIVISION CECIL SCOTT, et al Plaintiffs V. C. A. NO. B-84-1103-CA MONSANTO COMPANY Defendant PLAINTIFFS * NOTICE OF INTENTION TO TAKE ORAL DEPOSITION AND SUBPOENA DUCES TECUM TO: Defendant, MONSANTO COMPANY, by and through its counsel of record, Mr. Jonathan B. Shoebotham and Mr. Robert A. Hall, Woodard, Hall <5 Priirnn, 4700 Texas Commerce Tower, Houston, Texas 77002 Mr. Walter J. Crawford, Jr., Wells, Peyton, Beard, Greenberg, Hunt <5 Crawford, 624 Petroleum Bldg., P. 0. Box 3708, Beaumont, Texas 77704 Mr. John M. Johnson, Mr. Warren B. Lightfoot, Bradley, Arant, Rose & White, 1400 Park Place Tower, Birmingham, Alabama 35203 Mr. Michael R. Fruewald, Mr. Michael Rosiello, Ms. Anne C. McGown, Barnes & Thornburg, 1313 Merchants Bank Bldg. 11 South Meridian Street, Indianapolis, Indiana 46204 Pursuant to the provisions of Rule 30(b)(6) of the Federal Rules of Civil Procedure, the Plaintiffs in this cause hereby give notice to the Defendant that the oral deposition of Monsanto Company, Defendant, will be taken on the date and time and at the place hereinafter stated. Specifically, the Plaintiffs intend to take the oral deposition of the representative( s) designated by the 000C07 WATER PCB-SD0000012565 Defendant pursuant to Rule 30(b)(6) of the Federal Rules of Civil Procedure and charged with the responsibility for maintaining custody of the documents listed below and such designated representative(s) shall testify as to matters known or reasonably available to the Defendant as well as the custody of the herein designated records and their maintenance by Defendant: 1. The complete personnel file of Paul L. Wright; 2. Any bonuses, payment dividends or other consideration other than normal salary paid by Monsanto or anyone on its behalf to Paul L. Wright or his designee/assignee; 3. Paul L. Wright's job evaluation and/or reviews; 4. Any payment to or for the account of attorneys hired by or for Paul L. Wright in connection with matters pertaining to his employment at Industrial Bio-Test Laboratories, Inc. ("IBT"). See United States v. Keplinger. 776 F.2d 678 (7th Cir. 1985); 5. Any correspondence to or from Paul L. Wright; 6. Paul L. Wright's job duties and responsibilities as an employee of Monsanto both before and after he was employed by IBT; and, 7. The matters identified in the below listed subpoena duces tecum. The deposition will be taken at the offices of Gilpin, Pohl & Bennett, 1300 Post Oak Boulevard, Allied Bank Tower, 23rd Floor, Houston, Texas 77056 beginning at 1:30 p.m. on Friday, May 8, 1987, and shall continue until completed. This oral deposition will be taken before a certified court reporter. The testimony given during this oral deposition will be used as evidence in this matter, together 000C03 WATER PCB-SD0000012566 with the documents produced at this deposition. invited to attend and cross-examine. You are ' Pursuant to Rules 30(b)(5) and 34 of the Federal Rules of Civil Procedure, certain documents shall be produced by the designated representative(s) or custodian(s) of Monsanto Company (referred to herein as "Monsanto") at the commencement of the oral deposition or prior thereto by agreement of counsel. The originals and all drafts of the following requested documents shall be produced: 1. The complete personnel file of Paul L. Wright; 2. Any payment by Monsanto or anyone on its behalf to or for the benefit of Wright Paul L. Wright; 3. Any payment by or for Monsanto to attorneys for Paul L. Wright in connection with any criminal prosecution arising out of or in any way related to his employment at IBT; 4. All correspondence to or from Paul L. Wright; 5. All evaluations of Paul L. Wright's performance as an employee of Monsanto; 6. All documents pertaining to the termination of Paul L. Wright's Monsanto employment (both before being first, employed by IBT and as a result of or in connection with the criminal proceedings relating to IBT). See United States v. Keplinqer. 776 F.2d 678 (7th Cir. 1985); 7. All documents pertaining to Monsanto's efforts to monitor, supervise, keep abreast of or in any way observe the IBT criminal trial. See United States v. Keplinqer. 776 F.2d 678 (7th Cir. 1985); 8. Any report, analysis or investigation of IBT misconduct (actual or alleged) in connection with tests performed on Monsanto products (this request includes copies of government reports; independent investigations; internal IBT investigations; Monsanto authored or sponsored investigations; -3- 0000013 WATER PCB-SD0000012567 and, investigations) by industry groups, bodies or authorities or other private or public body) (IBT includes all of its employees and consultants). Unless otherwise specified herein, the documents to be produced include all documents in the possession of Monsanto from the date of the first production or manufacture of PCBs in any form by Monsanto to the present time. As used in this notice, the term "documents" means any printed, typewritten or handwritten matter, or reproduction thereof, of whatever character, in the possession, custody or control of a witness or his agents, representatives or employees, including without limitation, correspondence, contracts, memoranda, agreements, letters, brochures, reports, handwritten or typewritten notes, sound recordings or transcriptions thereof, computer print-outs, inter-company and intra-company communications, work papers, diaries, calendar pads, appointment books, ledgers, financial, statements, checks, bank drafts and writings of whatever kind and character, whether originals or reproductions, and whether in draft or final form, including copies bearing different markings or notations. By:--------------------------------------------------------------JOSEPH C. BLANKS -4- onoc.o WATER PCB-SD0000012568 OF COUNSEL: REAUD, MORGAN & QUINN 909 Laurel Avenue Beaumont, Texas 77701 (409) 838-9941 Michael A. Pohl David M. Lacey GILPIN, POHL & BENNETT 1300 Post Oak Blvd. Allied Bank Tower, 23rd Houston, Texas 77056 (713) 623-8800 Floor Thomas Henderson HENDERSON & GOLDBERG 1030 Fifth Avenue Pittsburgh, Pennsylvania 15219 Benton Musslewhite LAW OFFICES OF BENTON MUSSLEWHITE 609 Fannin, Suite 517 Houston, Texas 77002 (713) 222-2288 David S. McCrea McCREA & McCREA 119 S. Walnut Street Bloomington, Indiana (812) 336-4840 47402 ATTORNEYS FOR PLAINTIFFS, CECIL SCOTT, ET AL. -5- ooor,:i WATER PCB-SD0000012569 CERTIFICATE OF SERVICE I hereby certify that on the ______ day of , 1987, a duplicate original of the foregoing Plaintiffs' Notice of Intention to Take Oral Deposition was filed with the Clerk of the Court for the Eastern District of Texas, Beaumont Division. I also certify that a true and correct copy of this Notice was served upon counsel of record on the ______ day of ___________ 1987, by pessenger delivery and/or by placing same in the United States mail, certified mail, return receipt requested, postage prepaid and addressed as follows: Mr. Robert A. Hall Mr. Jonathan B. Shoebotham Woodard, Hall & Primm 4700 Texas Commerce Tower Houston, Texas 77002 Mr. Walter J. Crawford, Jr. Wells, Peyton, Beard, Greenberg, Hunt & Crawford P. 0. Box 3708 Beaumont, Texas 77704 Mr. John M. Johnson Mr. Warren B. Lightfoot Bradley, Arant, Rose & White 1400 Park Place Tower Birmingham, Alabama 35203 Mr. Michael R. Fruewald Mr. Michael Rosiello Ms. Anne C. McGown Barnes & Thornburg 13i3 Merchants Bank Bldg. 11 South Meridian Street Indianapolis, Indiana 46204 MAP:j ar/scottmon/054 JOSEPH C. BLANKS -6ocor:^ WATER PCB-SD0000012570 f V. E X H I B I T ( WATER PCB-SD0000012571 IN THE UNITED STATES DISTRICT COURT FOR THE EASTERN DISTRICT OF TEXAS BEAUMONT DIVISION CECIL SCOTT, ET AL. VS. MONSANTO COMPANY CIVIL ACTION NO. B-84-1103-CA PLAINTIFFS' NOTICE OF INTENTION TO TAKE ORAL DEPOSITION AND SUBPOENA DUCES TECUM TO: Defendant, MONSANTO COMPANY, by and through its attorneys of record Mr. Robert A. Hall, Mr. Jonathan B. Shoebotham, Woodard, Hall & Primm, 4700 Texas Commerce Tower, Houston, Texas 77002; .' Mr. Walter J. Crawford, Jr., Wells, Peyton, Beard, Greenberg, Hunt & Crawford, P. 0. Box 3709, Beaumont, Texas 77704; Mr. John M. Johnson, Mr. Warren B. Lightfoot, Bradley, Arant, Rose & White, 1400 Park Place Tower, Birmingham, Alabama 35203; and Mr. Michael R. Fruehwald, Mr. Michael Rosiello, and Ms. Anne C. McGown, Barnes & Thornburg, 1313 Merchants Bank Building, 11 South Meridian Street, Indianapolis, Indiana 46204. Pursuant to the provisions of Rule 30(b)(6) of the Federal Rules of Civil Procedure, the Plaintiffs in this cause hereby give NOTICE to the Defendant that the oral deposition of the Manager of Toxicology of MONSANTO COMPANY, Defendant, will be taken on the date and time and at the place hereinafter stated. Specifically, the Plaintiffs intend to take the oral deposition of the ' 2 DEPOSITION \ EXHIBIT 2l WATER PCB-SD0000012572 representative(s) designated by the Defendant pursuant to Rule 30(b) (6) of the Federal Rules of Civil Procedure and charged with the responsibility for maintaining custody of the documents listed below. The designated representative(s) shall testify as to: a) Matters known or reasonably available to the Defendants concerning the policies and procedures of Monsanto's Toxicology Department for the years 1965 through 1980; b) The current standards, procedures and policies of Monsanto's Toxicology Department; c) The duties and responsibilities of the Manager of Monsanto's Toxicology Department (i) 1965 through 1980 and (ii) currently; d) The organization of Monsanto's Toxicology Department (i) 1965 through 1980 and (ii) currently; e) The interrelationship between Monsanto's Toxicology Department and Monsanto's other departments and/or sections; f) The relationship between Monsanto's Toxicology Department and any outside testing facilities (i) 1965 through 1980 and (ii) currently; g) Any industry or government guidelines or regulations relating or pertaining to tests concerning the toxicological effects of Monsanto products on animals; h) Any participation by Monsanto's Toxicology Department in any effort by Monsanto to avoid or defer the ban of its PCB products; i) The day-to-day activities of the current Manager of Monsanto's Toxicology Department; and -2- OOOCM WATER PCB-SD0000012573 j) The documents designated in the accompanying Subpoena Duces Tecum. This deposition will be taken at the offices of Gilpin, Pohl & Bennett, 1300 Post Oak Boulevard, Allied Bank Tower, 23rd Floor, Houston, Texas 77056 beginning at 9:00 a.m. on Saturday, May 23, 1987 and shall continue until completed. This oral deposition will be taken before a certified court reporter. The testimony given during this oral deposition will be used as evidence in this matter, together with the documents produced at this deposition. You are invited to attend and cross-examine. Pursuant to Rules 30 (b) (5) and 34 of the Federal Rules of Civil Procedure, certain documents shall be produced by the designated representative(s) MONSANTO COMPANY (referred to herein as "Monsanto") at the commencement of the oral deposition. The originals and all drafts of the following requested.documents shall be produced: 1) Any manual, compilation or other writing setting forth or reciting the policy or procedures for the Monsanto Toxicology Department from 1965 to date; 2) Any document pertaining to Monsanto's standards for the feeding or care of animals used in connection with the testing of Monsanto products; 3) Any document pertaining to the care, skill or precision to be used (a) by Monsanto and (b) by any outside/independent testing laboratory in gathering, recording, maintaining and/or -3- 0000:5 WATER PCB-SD0000012574 reporting of data generated by or from animal studies; 4) Any document comparing or contrasting the results of IBT Aroclor tests with tests conducted by (a) Monsanto, (b) others on Monsanto's behalf and/or (c) independent or government researchers; 5) Any document regarding any investigation or analysis of those Aroclor tests conducted by IBT for Monsanto; 6) Any document regarding or otherwise pertaining to trips (a) by Monsanto employees or representatives (i) to the offices of IBT (ii) to other locations where IBT representatives were met or (b) by IBT employees or representatives (i) to the offices of Monsanto or (ii) to locations where IBT and Monsanto employees or representatives met; 7) All literature regarding PCBs which was supplied or made available to Monsanto's experts, or consultants; 8) All Electrical Power Research Institute reports, articles, papers, etc., pertaining or relating to PCBs; 9) Any list or summary identifying (a) which IBT studies were redone by or for Monsanto and (b) the variances or similarities between the Monsanto tests conducted by IBT and those redone by or for Monsanto; 10) Any industry (and/or government) guidelines, standards or suggested guidelines/standards applicable to toxicological studies from 1960 to date (e.g. Aroclor studies on rats, beagle dogs, leghorn hens and/or fish) including any variations thereof from their inception to date; 11) Any document setting forth or otherwise stating Monsanto's standard of care regarding the following data in connection with toxicological tests (gathering, recording, maintaining) ; a) blood data; -4- Q00C1G WATER PCB-SD0000012575 b) urine analysis; c) body weight; d) feeding data; e) autopsy (procedures and results) ; f) viscera; g) microscopic analysis; h) organ analysis; i) death of test animals; j) substitution of animals; k) loss of animals due to escape or confusion of identity. 12) All correspondence between IBT and Dr. Paul Wright while Dr. Wright was employed by Monsanto; 13) Any document evidencing or suggesting that Monsanto corrected, changed, altered or otherwise amended (a) any report of a test performed by IBT for Monsanto and (b) any paper published or to be published regarding IBT's study of any Aroclor product; 14) Any document (a) evidencing or otherwise pertaining to suggestion by Monsanto as to the "results" to be achieved by IBT in connection with the tests by IBT of Monsanto products, and (b) Monsanto's policy in regarding thereto (i) 1965 to 1980 and (ii) currently; and 15) Those documents relating to items a through j first above listed. As used in this notice, the term "documents" means any printed, typewritten or handwritten matter, or reproduction thereof, of whatever character, in the possession, custody or control of a witness or his agents, representatives or employees, including without limitation, correspondence, contracts, memoranda, agreements, letters, brochures, reports, handwritten or typewritten notes, sound recordings or transcriptions thereof, computer print-outs, 000017 WATER PCB-SD0000012576 inter-company and intra-company communications, work papers, diaries, calendar pads, appointment books, ledgers, financial statements, checks, bank drafts and writings of whatever kind and character, whether originals or reproductions, and whether in draft or final form, including copies bearing different markings or notations. "Aroclor" shall mean any Monsanto product containing PCBs or their contaminants. "Animal" shall mean any animal, mammal, reptile, bird, or fish used in connection with any study. "PCB" means polychlorinated biphenyl and its contaminants and derivatives. "Toxicology Department" means any department or section responsible for investigating or determining the possible toxicological effects of exposure to products manufactured by Monsanto Company. ' Respectfully submitted. By: Michael A. Pohl OF COUNSEL: David M. Lacey GILPIN, POHL & BENNETT 1300 Post Oak Boulevard Allied Bank Tower, 23rd Houston, Texas 77056 (713) 623-8800 Floor OOOC^G WATER PCB-SD0000012577 REAUD, MORGAN & QUINN 909 Laurel Avenue. Beaumont, Texas 77701 (409) 838-9941 Thomas Henderson HENDERSON & GOLDBERG 1030 Fifth Avenue Pittsburgh, Pennsylvania (412) 471-3980 15219 Benton Musslewhite LAW OFFICES OF BENTON MUSSLEWHITE 609 Fannin, Suite 517 Houston, Texas 77002 (713) 222-2288 David S. McCrea McCREA & McCREA 119 S. Walnut Street Bloomington, Indiana (812) 336-4840 47402 ATTORNEYS FOR PLAINTIFFS, CECIL SCOTT, ET AL. CERTIFICATE OF SERVICE I hereby certify that on the ______ day of , 1987 , a true and correct copy of the above Notice of Intent to Take Oral Deposition and Subpoena Duces Tecum was served upon counsel of record either by messenger or by placing same in the United States mail, certified mail, return receipt requested, postage prepaid and addressed as follows: Mr. Robert A. Hall Mr. Jonathan B. Shoebotham Woodard, Hall & Primm 4700 Texas Commerce Tower Houston, Texas 77002 Mr. Walter J. Crawford, Jr. Wells, Peyton, Beard, Greenberg, Hunt & Crawford, P. 0. Box 3708 oooc:o WATER PCB-SD0000012578 Beaumont, Texas 77704 Mr. John M. Johnson Mr.'Warren B. Lightfoot Bradley, Arant, Rose & White 1400 Park Place Tower Birmingham, Alabama 35203 Mr. Michael R. Fruehwald Mr. Michael Rosiello Ms. Anne C. McGown Barnes & Thornburg 1313 Merchants Bank Building 11 South Meridian Street Indianapolis, Indiana 46204 depntc0 2/txt8 6551 Michael A. Pohl (Moc;:o WATER PCB-SD0000012579 I V ~ ~v ''l E X H I B I T WATER PCB-SD0000012580 TUz VUI: 650 26J0717 <409) 038-9941 EBBS, MM6IN V MINN LAVTZBS 909 Laurel ____ Beaumont Texa* 77701 itmrkl: MCI JBLAHES HCI ID: 263 0717 May 13, 1067 Walter Crawford WELLS, PEYTOH, BEARD, ICEBERG, HUNT & CRAVEORD 624 Petroleum Building BEAUMONT, TEXAS 77704-3706 Re: CecU Scott v. Monsanto Company, B-64-1103-CA Dear Walter: In order to minimize the burden on you of complying with Plaintiffs' requests for item 6 on IBT related documents --- which requests first came December 31, 1966, were renewed via 30(b)(6) notices April 3,1967, and most recently, April 29, 1967 -- I repeat my verbal agreement with you of this morning that item 6 will at this time be satisfied if you provide each summary report regarding the analyses of, investigations of, and other detailed reports on IBT misconduct (actual or alleged) in connection with tests performed on Monsanto products. I understand that IBT may have done as many as 650 separate studies for Monsanto on some 33 products. I also understand that third parties may have done some 330 study validations on IBT studies of about 30 Monsanto products. It is not clear whether the study validations were done before or after the IBT misconduct came to light. At this time, we do not insist on having the 350 study validations. We do, however, want to know the results of such validations, that is, whether or not the IBT studies were or were not found to be valid, and if not, why not. 2 DEPOSITION ? EXHIBIT fLev/as KKs-y 1 r-0.-7.y-7 u6f< GQOCm WATER PCB-SD0000012581 As concerns tills particular request reports about IBT misconduct -- I suspect that Monsanto has some summary reports about the tests, the validations. Independent Investigations, Internal IBT Investigations, and even governmental Investigations or reports. I suggest that you start with the reports presented to the highest level of Monsanto management and work your way downward until you reach the detailed study validations, stopping just short of them. Surely this will simplify your quest, and with the use of the wonderful Monsanto PCB Index and document retrieval system and data base, you can gather this handful of documents together In a matter of days --- having already had months since the Initial requests to Identify the dangerous documents. Thank you for your ongoing help in this matter. ooor/j-~ WATER PCB-SD0000012582 u jx x I WATER PCB-SD0000012583 Monsanto <% * ! 9m )( i ttO"*** George Roush, Jr., M. D. ATI wtjicr , September 17# 1975 MONTHLY COMMENTS - MEDICAL DEPARTMENT TO : Corporate Administrative Committee CONFIDENTIAL r<I* State and federal agency activities related to PCB's continue to require Medical Department attention to support business group efforts. Recent public hearings in Wisconsin served as a forum for Mr. Schweitzer, of EPA's Toxic Substances Office,to make his pitch again in support of the Toxic Substances Control Act now being considered in Congress. Between now and the end of November, FDA, EPA, and NIOSB will conduct hearings on various aspects of the'PCB's in foods and the environ ment. Questions from outside 'Monsanto about the long-term health effects of chemicals used in two Monsanto plants on the workers in the plants have required organization of epidemiological studies. These require acquisition of data on individual work assignments within the plant over long periods of time, collection of death certificates of deceased employees, and comparison of causes of death with frequency of cause to some segment of the general population. We will have to do much more epidemiology in the future although clear cut conclusions are almost impossible. GR/ln i DEPOSITION I? EXHIBIT S" i S'ST-gn uok. SCM 019722 0000^3 WATER PCB-SD0000012584 WATER PCB-SD0000012585 'Monsanto ' r*o (< * tOCATiom Medical Department A2SA October 13*, 1971 AROCLOi^ 1260 te R. E. Kelly M. M. Johnaon E P. Wheeler TO PILE !. MV IMI I spoke with: Tel: Renate D. Kimbrough, M.D. Pathologist EPA Atlanta Tox. Branch 4770 Buford Highway Chamblee, Georgia 303*11 (404) 633-3311, ext. 5218 today regarding her obaervatlon of bladder tumors In rata fed AROCLOR 1260. The observations of Voa and Koenan (Toxicol, and Applied Pharmacol. 17, 656-668, 1970) on polychlorinated biphenyls led them to undertake rat reproduction studies with AROCLOR 1254 (Lot AX38) and 1260 (Lot AK3). The materials were obtained from Glasgow at PDA. Their primary findings have been lea Iona In the liver, and Dr. Kimbrough plans to present a paper on the liver lesions at the 1972 spring meeting of the Society of Toxicology. In addition, she haa seen 2 lesions In the bladders of rats fed AROCLOR 1260. The first occurred In a female which died after 6 months on test. This was dlagnoaed as a malignant anaplastic carcinoma of the bladder. The second was In a male killed after 8 months on test. The first diagnosis was of epithelial hyperplasia. Sections of both bladders were sent to NCI for diagnosis to Drs. Strauss (7) and Katherine Snell. The diagnosis of carcinoma in the female was confirmed. The lesion In the male was not resolved. Some pathologists believe It Is a carcinoma, others believe it is -.hyperplasia. Dr. Kimbrough stated that they were trying to analyze the AROCLOR 1260 sample for Impurities, but were having some difficulty with their equipment. I Indicated that we would try to track down material from the sample that they had received. In the event that we cannot locate this lot, we probably can get some material back through Thomas B. Gaines, Supervisory Research Pharmacologist, at the sane location as Dr. Kimbrough. If we have an analysis of this lot, we should make the results known to Dr. Kimbrough. Dr. Kimbrough said that they had observed porphyria In some rats with both compounds. She also Indicated that they were contem plating doing a 2 year feeding study with larger nunbera of rats to Investigate the problem of bladder tumors. She is per plexed by the bladder tumors, and she Is not emphasizing them in her discussions. However, she has mentioned her findings when PCB's have been discussed at various Interagency meetings. This Is apparrently how Weissburg learned about them. (cont'd.) H DEPOSITION t exhibit j^Wcy.sftr/K- (r 3") iOA* lOOOitt WATER PCB-SD0000012586 FILS October 13, 1971 Page - 2 - As a final note, Dr. Kimbrough expressed concern over whether PCB's presented an additional hazard to the employees who manufacture it. 1 told here that because of the chloracne and liver hazards, there had been medical supervision of the employees. However, I would raise this point with Drs. Kelly and Johnson for their review. , . /blea eorge\JJ. Levlnskaa N >i } SCK ooor; WATER PCB-SD0000012587 U iX X ( \ I B I T / I WATER PCB-SD0000012588 October 28, 1971 Or. Moreno L. Keplinger Induetrial Bio-Teet Laboratories, Inc. 1810 Frontage Road Horthbrcnk, Illinois 60062 Dear Kept I spoke with Or. Senate Kimbrough 0f the S?A In Chaablee, Georgia regarding our proposed visit to her to discuss bladder pathology In rats fed this product. Z Indicated . to her that we expected to have all of the available bladders examined by the latter part of Voveaber, and that we would confirm a specific data beforehand. The express purpose of the visit Is to exchange Information, l.e., view the slides she has, and let her see any slides that Lon turns up which aay be of interest. This was agreeable to her. She suggested that we might bring extra slides for referral to KCI, if we so desired. After Don has finished his evaluation of the bladder sections, and after you and he have had a change to review his schedule, I would appreciate a c-pple'of dates that would be convenient for a trip to Chamblee so that Z aay schedule a visit there. Xlndest personal regards. George J. Levlnskas, Ph. 0. Mgr., Product Svaluatloa OJVbks cct Dr. Don Gordon Mr. V. B. Pspegeorge Mr. K. P. Wheeler i DEPOSITION 1 i EXHIBIT I lLev/v&.fte- 7 I tr-3-7- X~) ooc SCM 023586 OOOG^G WATER PCB-SD0000012589 ID X X V I B I T WATER PCB-SD0000012590 WATER PCB-SD0000012591 U iX X i i i WATER_PCB-SDOOOO012592 ^Endeacd' cd'tUo* rat :ttaior>;lit Hite' A*-roeelWor otudlr. +' Tho final ^ura4,t%'iUia?9ab\Sintlcn ayS'ahr' oliiiiOy;^fajmdlna-; . . in its mnalnc'onl.'h&'nt^iutaoid providcddoscripllw terms for thoaa s> lo-lon on he-cnl'cr'11-lJtli. and with .toUmax..you for tin use of your . notorial. Ho laslona will ta'ldontiricdvlUi.iiny 'epoelfic ccapouad, now --or l>a- *tiin-- *Cutwo..*?.factI^nolocted tto'elldaa bllarlly "to'roprrrent" have you uUnnd,i'.ll> you likei\ It la balng bool la Un.SImratoa Motor Hotel lu iiU.tor Sprlmi, frrm H:30'a`ii.V ixjcrmbor. 11- --iSsOO ntva .. . . w--- ..-' ' i\ .,- . . tba tlca ua tbla Aroclor stoJy. uut y.aclilulu nutotollcatlena ltanra' t ; teuton qulta ovnrvlKilaiat'."' joou aa tlw epcrlall<tolna uu conflated, I should bo'uUa.to write tin patliiUotf .doccrlpUcns and labulatlona'. , ;;' *^'^- 1 WATER PCB-SD0000012593 WATER PCB-SDOOOO012594 UJXX WATER PCB-SD0000012595 P.R.Johonnsen - A2SC December 6, 1974 AROCLOR 1254 0. Roush A2SA Dirc 111:-74` E. P. Wheeler - A2SA W. B. Papajeor^e - E2CK / / S In a recent lphone conversation between Dr. R. Kimbrough (CDO) and 0. J. Lcvinslcas, Dr. Kimbrough expressed the telief that KCI had recently contracted out a carcinogenicity study Involving AROCLOR 1254. Consequently, I made a call to Dr. Sidney Siegel at IICI. Dr. Siegel Indicated that he had no knowledge of an outside contract with ARCCLCR 1254 and further stated he doubted there was one. He did tell me that AROCLOR 1254 has been under test at IICI in their bicassa.y operation program since September 1972. He reported that the experimental part of the study load "been completed aj^of October, 1974. His expectations are that the necessary histopathology'Trt.irTje completed within the next 6 months, at uhich time a meeting could be set up for discussion oTTfe test results. '----------------------- . - A quick chec!: of the April 3* 1974 list of chemicals being tested . under the IICI carcinogenesis program did not reveal that AROCLOR 12^4 was on test. However, the NCI list does show C02664 BIPHENYL, CHLORO /bkp osi*8 sc* WATER PCB-SD0000012596 UJXX I \ B 1 T n WATER PCB-SD0000012597 j ) OCPARTMCNT OF HKALTH, KDUCATION,>NO WKLFARt PUCUC HCALTH SCftVICC CCKTKft PON OillAIC COW!**. ATT-AHTA, QCOAOiA MUl rn i<h uuil December 6, 1974 f l' Dr. George La-ciakes Monsanto Industrial Gbesdcale Coapasy 900 I. Lindbergh Boulerard SC. Louie, Missouri 63164 Deer George: I ea enclosing e brief outline of the experiment Aroelor 12-72-A for your Information. A copy of Bob Squires letter la eleo enclosed^ Sincerely yours, &&oCZ!tr Senate D. Kiibrouih, M.D. Toxicology Branch Enclosure WATER PCB-SD0000012598 LUXX I II \ I B I T WATER PCB-SDOOOO012599 . *;v aSsT,; -.^ : :- P2<Jws*i '. ISiteSt^ 3 cV' $jE#SI??5te- -i-v* *$`5* ~- ' ` *1. `` *J- , -.'*.2 " r^sj As s follow-up to-our phone conversation yesterday, X aa .. sending you copies'ofthe lnformtlon oo Aroclor 1260 study which was sent to. be by Dr* Klabrough. . You nay find this i; Informative In preparing.for your subsequent'discussion with , ^ When you have had a chance. to review yourschedule, would you . _ kindly let ae know What days may be convenient for a aeeting. `V This probably will be In the Washington area and X anticipate ; that you, Renate Klabrough, Bob Squires and I will attend. - ..,.V * - t ii. ..,**// . \17--,v1\. r*'V.'. y f -C. * \ ..*. ?. '-?j.;' v * r "'z* ," - - - DEPOSITION EXHIBIT VAWSflJ- )Q uo/c : < 0000312 n WATER_PCB-SD0000012600 SOI 70* fmiCAIIOH o* CI1CTJIAT10H Aroclor 12-72-A Animals Random brad ftaala Sharman aCrala rata raarad uadar tpaelfle pathogan-fram conditions vara obtained froa tha SCSC farm at Lavrencavilla, Ga. Aroclor 1260 Aroclor 1260 (Lot number AX-3) vaa supplied bp Monsanto Industrial Chaalcala Co., St. Louis, Mo. iI Statistics Tha students t-tast vaa usad for comparing body valghts and weight gain. Methods . Pour hundred 21-26 dap-old veanllng female rats, weighing 46-97 g, vara distributed into 2 groups of 200 animals each according to a table of random numbers. Each animal, was valghad and given an Individual ear 1 tag number. Tha animals vara housed 10 rats par cage, food and vater vara provided ad libitum. Two hundred animals vera fed tha control diet of ground Purina laboratory chow; tha taat animals vera fed the saaa diet fortified with 100 ppm Aroclor 1260. Aroclor 1260 vas Incorporated Into tha diet bp dissolving 5.2 g In ethpl ether than adding this to 100 g corn starch and allowing tha ether to evaporate. Tha PCS-cornstarch mixture vas blended with 345 g ground ehov, than pulverised In a mortar and diluted with additional ground chow in a bakare mixer to give a 1000 ppm concentrate. Tha concentrate vas further diluted to obtain 50 kg of 100 ppm diet. Tha diet vas prepared every 10-14 daps. Random samples from tha final mixes along with samples of control chov vara taken at regular Intervals to determine PCS levels la tha control chov and to verlfp tha 100 ppm fortification level. Ground laboratory chow vas also checked at intervals for aflotoxln at tha dls trice of flea of ehe Food and Drug Administration in Rev Orleans. The combined body weight of rats In each cage vas recorded weekly until tha rats vera 6 months old, bi-veekip until 12 months old, and monthly thereafter. Individual velghts were recorded only at the onset of the experiment end at daath or sacrlflca. Tha animals vera obaarved SCM 022811 nnnc.p/A WATER PCB-SD0000012601 I Axoclor 12-72-1 - Fsge 2 briefly each day tad snimals exhibiting debilitating signs or large tumors ware removed from the group cag tad housed individually. At each weighing the animals were examined individually and abnormalities noted, food consumption vaa measured on all rata during the first two weeks of the experiment, then during weeks 3, 8, 11 and 20 and every 12 weeks thereafter. Antopsies were performed on all rets that died, lets that were sacrificed were killed under ether anesthesia by severing the vena cars. Tissues were fixed in buffered formalin and stained with hematoxylin and eosln. Additional special stains will be conducted. laaulta During the course of the study 4 animals is the test group and 8 controls were sacrificed prior to the final kill. These animals had either large tumors which had ulcerated or hindered movement, or the mlasl appeared moribund. In all, 14 animals la the test group died; 17 animals in the control group died. One animal.was accidently killed in each group early in the study. Ifo definite dose related signs of toxicity were observed In the teet animals throughout the study. Comparative curves of body weight gain and food consumption on the basis of body weight, es well as K1 intake of the test group are given in Figure 1. A slight decline in the rate of weight gala of the teat group cohered to the control group began about 3 months after onset of the experiment. Mean final body weights were 420 g (S.D. 72, S.Z. 3.4) for the control group and 392 g (S.D. 62, S.Z. 4.6) in the test group; the difference was statistically significant (p<0.001). Food consumption (ga/rst/dsy) was comparable in both groups throughout the study. Kean weight gain was 330 g (S.D. 70, S.Z. 3.3) and 323 g (S.D. 60, S.Z. 4.3) for the control end test.gToups, respectively; the difference in weight gain was also statistically significant (p<0.001). FCB intake declined from 11.6 mg/kg/dsy during the first week of exposure to 6.1 mg/kg/dsy at 3 months of exposure and to 4.3 mgAg/day at 20 months. A 6 g mean weight loss occurred in both control and test groups during the 6 weeks prior to the final kill. 1 $CH 0228 V1* QOO( WATER PCB-SD0000012602 Preliminary of Microscopic findings (lisbrou*h) Please Hoc*: Sections of All Tissues Listed Vers Rot Aval lib Is for All Animals Timor True Controls (Tissue of one rat autoLysed) Experimental Rats Mubuiy Gland: fibroadenoma Adenocarcinoma 19 7 14 0 Pituitary: Chromophobe Adenoma Csreinoas Pituitary 38 0 30 1 Uterus: Endometrial Polyp Endometrial Sarcoma 20 2 23 - 3 and 1 adenocarcinoma Thyroid: Medullary Call Tuaors (parafollicular) . 26 18 Liver: Modular HypsrplaaIs 3 (8188,8192,8232) 7 . Nodular Hyperplasia and Hepatomas 1 (8231) 134 Nodular Hyperplasia* Hepatomas, and Hepatocellu lar carcinomas 0 Adenoflbroels 0 14 4 Bladder: Papilloma 1 (8317) 0 Is addition, a nusfaer of ocher occasional tumors were found. SCM 02281 000C35 WATER PCB-SD0000012603 Praliadnary Liat of Microscopic Pladlnga - Paga 2 Out of thia group t 4 control rata and 4 rata la tha axparlaaatal group vara aacrlflead aarly baeauaa thay vara tick* All tumora of thaaa aalaala, outalda of tha llrar laalcma, ara laeludad la tha praeaadlag list. Tha llvar laaioaa naad to ba addad. Tha aalaala ara aa follovai Controla . txparlaantal 8194 8131 8121 8234 8331 8347 8413 8317 SCH 022816 WATER PCB-SD0000012604 500 400T Figure 1 BODY WEIGHT 5 ,o X u l/> I J c O co Body Weight in gm 300H 200H 100H Control 100 ppm Aroclor 1260 I vi i i rri i i i "i-r11 i r i-r > tt i i i i i 1 2 3 4 3 6 7 6 9 1011 12 13 14 15 16 17 10 192021 2223 24 Months I I I I I Ij I 1 I I\ i WATER PCB-SD0000012605 ecuooo Figure 2 FOOD INTAKE AND PCB CONSUMPTION /~\ <> rrOo fO\J Xoin \Ci n i. / -i WATER PCB-SD0000012606 xxm ! II i!1 ii \ I B I T # Ii WATER PCB-SD0000012607 % i ;w V ***' ..W S\ Aw . AltOCLCR 126o: Meeting at MX, J11M17 31, 1975 044 The purpose of this aeetlng u to rtrlw sections of liver tissue from Or. Kimbrough's 2-year study in which female rets were fed 100 ppa of AROCLOR 1260. Ort. Kimbrough end Rqulre had studied end elesslfled the findings. Or. Levitt, either a visiting fellow or a post doctoral trainee did not participate to any great extent. He was Mentioned as having a good knowledge of liver. Dr. Rlenter had viewed the slides froa our 2-year study In which rats of both sexes had been fed 100 ppm, 10 ppa or 1 ppa of AROCLOR 1260. Subsequently, be and Or. Gordon reviewed sections of liver froa all anlaala in this study. Three conclusions can be drawn. 1. In our earlier study, the severity of liver lesions war; greater In fearles than In salas. ?. To a large extent, substantially the saae type of leslonr were observed In both studies except that- the lenlons eeeaed to be aore advanced In flab rough's study. Although thore was tone variation In terminology, the findings were reasonably close. 3, There were definite liver adenocarelnoaas In Kimbrough's study. Or. Richter expressed the view later that 2 ani mals in our study approached the type of lesloo brough had observed, but there was agreement by Drs. Gor don and Ric.iter that Or. Kimbrough's rats had developed a lesion which they had not observed In our eerller study with AROCLOR 1260. If. Drs. Gordon or Richter feel that I have not suanarlxed this meeting correctly, or If they desire to aaend or expand ay remarks, I Invite them to let m know. jj Akp 2/3/75 -fccorge J. Levlnskas 0^0033 op WATER PCB-SD0000012608 XUJ T I B WATER_PCB-SDOOOO012609 ii /'^r^^/tl8tQsTro'0r^XUbrodiEb*i>S;`)rB<u>< "' - >'Tmts were fedlOO Jpfit* ' ` _ ii,vjs! ` __ v ','' findings.; tDr.^tiervltt;;^lib*r,^':;Tlltin ttHow ok>t '*;irdoctoral trainee--did-<abt ' ' '` 'W `w'&e "WS &--SZ-. ' ''? which rats of both sexes.-badibeen.red 100 ppa,trio, fipii or. '^Wi.l ppm or AB0CU>K126O 3* Subsequently,'he and-IT^Oorton-SP/:- ;; 'T^&seyiewed toc^oM.;cr;livfrjtt^-<3jL.Mrt*TiyjUa%thifjtad^r.a * vW -." . ' ., .". v- ' '` tmt; la fearlc^twa''la. v.. -- ?. Toalarge extent,7 substantially the.same typeof lesions were observed In both atediet except, tbat-tbe ileelens'v.-.v seemed to be more advancedln Kinb rough' a stndy.> Although there ' " " ` ' .................................... ....... -. . v were . .*>>' \ . . . -',r*,.-v'v` ..' -i study. Dr. Richter.,expressed the view.later that 2 end-f. ala In our study'approached the type;of lesioo 'Sla- .r ^ J4<' ~ brough had observed, but there was agreement by;Dra. Oor-,^ > don and Rleater that Dr.? Klabrough a rats had .developed ` `iTg.-' lesion whlch'they had not'observed'In our.earlier study Vv.; with M0CL0K.-126O.' \' * * -vL.-> i If; ;j)rs.^Gordon" or; Richter feelvtbatX'have hot 'Sunasrlsed this r v ''meeting correctly, or If they'desire .to anend orexpand my ,s .. " DEPOSITION EXHIBIT |6WV5^VtS-/^| ^7-?7 - COg^ r WATER_PCB-SD0000012610 xxm ! I B I T WATER PCB-SD0000012611 k TOXICQlOaT (NVIHQNMCNTAi CICNCKS CMCMIITIT AUNT CIINCCI MKOieAk seiKMcta !}tukAtf/u&L BIO -TEST JtcrfytfjObbyuAb, JJnc. 1810 S*ONTAG8 *OAO NORTH8ROOK. ILLINOIS 60062 March 24, 1975 ** COOK Jil rCLCMONC I7MOJO George J. Levinskas, Ph. D. Monsanto Company 800 N. Lindbergh Blvd. St. Louis, Missouri 63166 Dear Or. Levinskas: . I am enclosing the trip reports prepared by Or. Richter and myself regarding our meeting at N. C. I. on January 31, 1975 with Ors. Kimbrough and Squire on Aroclor 1260 (IBT No. 622-07298). . .. _, . . . ' . *. t The micro-slides of the liver sections from all these animals and' those from Aroclor 1242 and 1254 will be forwarded today, under separate cover. We are also mailing the reports relevant to these studies today. The charges for this additional work will be billed under a new Number (IBT No. 641-06672) instead of the previous number. Sincerely O. . Gordon, D. V. M., Ph. D Section Head, Pathology Dept. DEG:hb Enclosure DEPOSITION } EXHIBIT | LGVfh)<j#s- /S'! SCM 023632 WATER PCB-SD0000012612 February 2, 1975 Trip Report On January 31, 1975; Ora. Richter, Levinskas and myself met with Drs. Squire and Kimbrough at the National Cancer Institute in Bethesda, Maryland. The purpose of this meeting was to review the slides and data from a 23 month oral feeding study with Aroclor 1260 in rats that was con ducted by Or. Kimbrough while at the E. P.A. In her study, 200 female rats (Sherman strain) were fed 100 ppm of Aroclor 1260 in the diet for 21 months, while 200 female rata of the same strain served as controls and were fad Purina Rat Chow. The Sherman Rat is a random-bred animal derived from the Osborn Mendel Strain. The animals were 21-26 days of ' age when the study began, and were sacrificed after 23 months on the study. Therefore, there was a 2 month recovery period at the end of the study in which the test material was removed from the diet. The details of the ex perimental design, procedures and results that were forwarded to Or. Lavinskas are attached. In summary. Dr. Kimbrough found a rather high inci dence of hyperplastic (nodular hyperplasia) and neoplastic (hepatomas, car cinomas) lesions in the liver of the test animals. The slides were subsequently reviewed by Dr. Robert Squire, Head of the'Tumor Pathology Brlnch of The National Cancer Institute, who concurred with her findings, although, the ' . terminology he used in classification of the lesions varied slightly from Dr. Kimbrough's. In this regard, it was his impression that all hyperplastic nodules should be regarded as "pro-cancerous lesions" and the term "hepatomas" SCH 023633 00QC.J3 WATER PCB-SD0000012613 Trip Report 2 bo delated and re-classi/led as carcinomas or neoplastic nodule*. Tbi* classification system was based on & Liver Tumor Workshop that was held on Dec. 11-13, 1974 in Silver Springs, Maryland which was attended by a small group of pathologists from the regulatory agencies, research labora tories and Industry. In view of Dr. Kimbrough's findings, additional sections of liver from a 2-Tear Oral Feeding Study With Aroclor 1260,. in rats, conducted at Indus trial Bio-Test were processed and evaluated. In this study, there was a 3, 8 and 14 month interim sacrifice which enabled one to determine the ap proximate time of onset of the liver changes. No interim sacrifices were, conducted la Dr. Kimbrough's study. The additional flides prepared at . . Bio-Test were examined by Dr. Richter and a tabulation of the liver findings are attached. These slides were reviewed by Drs. Kimbrough and Squire at our meeting. Although there was some disagreement on the terminology used in classification of the lesions. It was apparent that the incidence and severity of liver lesions (hyperplasia, nodular hyperplasia and neoplastic changes - carcinomas) was greater in Dr. Kimbrough's study when compared with the results of the Bio-Test Study, even though the same level of test material was administered in the diet. There are two possible explanations for this difference: 1.) The strain of rat. Whan questioned. Dr. Kimbrough had no informa tion on the spontaneous incidence of hyperplastic or neoplastic liver . lesions in the Sherman strain. However, the Incidence of spontaneous liver cell hyperplasia would appear to be extremely low since this finding was reported in only 4 controls. The incidence of liver hyperplasia among control rats of the Bio-Test study was also very low. The Incidence of spontaneous hyperplastic liver nodules reached 20f* in the Fischer Rat 000041 WATER PCB-SD0000012614 Trip Report 3 and In loma of the other strains. 2.) Sex difference* On the basis of our atudiee and thoae of other in ' read gator a in rata and mice, it waa found that the incidence of hyper* plaatic and neopiaatic liver lealone are much higher in femalee. The reaulta of the liver tumor workahop are to be publiahed in the near future and will include the participante* The viewa of thia group, regarding claaaificatioa of hyperplaatic and neopiaatic liver laaiona is based on the pathogeneaia of auch leaione in rats that have been Induced by known carcinogens auch as the nitroaoaminea and 2. - acetylagdnoflorine (2-AAF). The adaption of this terminology and claaaification ayetem by the scientific community will have wideapread implicationa regarding aaaeaament of aafety of chemicals and drugs currently on teat and thoae previously studied. Attached are Dr. Richter's comments in regard to Dr. Kimbrough's findings. . Donovan E. Gordon, D. V. M., Ph. D. DlpL., Am. College Vet. Path. SCM 023635 0000-15 WATER PCB-SD0000012615 Or. Donovan Gordon Indue trial Bio-Teat Laboratories February 3, 1975 Z wish to summarize my observations of the lesions seen in Or. Kimbrough's study of Aroclor 1260 end the comparison of diagnoses with Or. Kimbrough and Dr. Squire. 1. ) The criteria that I used for diagnosis of the lesions are similar to what they used. 2. ) My evaluation tends to be a little more conservative than theirs. For example: they Would call some of my hepatomas careinoma but with some question. 3. ) Therefore, if we both read the same slides there might be a little variation in numbers of lesions in the different categories but no major difference. 4.) Dr. Squire and Or. Kimbrough are using a new and revised terminology for the categories and list them as follows: My Terminology (Classical Use) Dr. Squire's (Revised Terminology) Focal Hypertrophy Cellular Alteration Nodular Neoplasia or Neoplastic Nodule Carcinoma Carcinoma 5.) However, the lesions in Or. Kimbrough's study were more severe those in the Bio-Test study. The lesions that she and Or. Squire are calling carcinoma are also carcinomas by my criteria. I would WATER PCB-SD0000012616 conclude from in examination of their material that Or. Kimbrough's study demonstrated carcinogenicity. 44*/^. kUA&i Ward R. Richter, D. V. M. Dipl. Am. College Vet. Path. SCM 023637 000047 WATER PCB-SD0000012617 LUXX I B I T r WATER_PCB-SDOOOO012618 *7 25, 1970 Otla E. Fancher, Ph.S. Director Industrial Bio-Teat Laboratories, Inc. - 1810 Prontage Road Northbrook. Illinois 60062 Osar Otis: * This 1st tar will authoriss you to procssd with a study with ths laghorn chicksns with ths Aroclor 1242. lot number AL-55. This should dupliesto ths study you did previously with Aroclor 1242. lot number AX-255, axeapt in this instance you haws suggested using dietary levels of 2. 4. and 8 ppa. This sample of Aroclor represents our current regu lar production which involves some clean-up insti tuted since the previous sample was aade available ' to you. Ve would hope that we might find a higher no effect" level with this sample as compared to the previous work. - As soon as suitable reference standards are available, the research laboratory in St. Louis will acre care fully characterize the two samples in terms of possi ble minute amounts of contaminants. Best personal regards. Sincerely, Elmer P. Vheeler Manager, Environmental Health SCM 023565 QOODir} WATER PCB-SD0000012619 LUXX ]I I B I T # i WATER PCB-SD0000012620 fCB TO*iCOLOV KNvmoNMCirTAL sciences CMCMISTWV *-** sciences mcmcm sciences . Jn&uMai BIO - TEST J!abo*aiMie&, Sno. - 1B10 ^RONfAGH ROAD NORTHBROOK. ILLINOIS 60042 September 30, 1971 anca eooc SI* niUNONf lffl< Dr. George J. Levinskas Manager, Product Evaluation Monsanto Company 800 S. Lindbergh Boulevard St. Louie, Mieeouri 63166 Dear George: Enclosed are the. correction pages for the three teratology studies with Arodor 1242, Arodor 1254 and Arodor 1260. I discussed die interpretation of die renal caudal ectopia, found with Arodor 1254, with Dr. Keplinger. He is in agreement with the interpretation that this is probably not a specific teratogenic effect but a general indication of the toadcity of the materiaL hi reviewing the results from the three generation study there appears to be a reduction in both 5 day and 21 day pup survival in die group fed die highest level (100 ppm) of Arodor 1254. This was not observed in the other Aroder groups or in groups fed 1 or 10 ppm, again suggesting that die level of Arodor had a general toxic effect. As I pointed out in our phone conversation, die small differences observed la sex ratios are not significant. A statement to that effect has been inserted. If you have further questions, contact me at any time. PLW :1am Enclosures Sincerely yours, Paul L. Wright Section Head, Toxicdogy r- deposition f EXHIBIT iSLEV/A)St:#S-n\ cO<. onor'0 SCM 02350^ WATER PCB-SD0000012621 U IX X \ l B I f / t WATER PCB-SD0000012622 April 28, 1979 * deposition I j exhibit !i S*AO ^7 OJ/cl Dr. Otis Fancher 624 H. Abrego Drive Oreen Talley, Arisons 85614 . Door Otisi Z an aahaaed that I have heenlso slow; in.-reviewiag the aanuacripta and sending you our eosawnts. l' guess Xsp's sending as the chronic rat and dominant lethal and chicken study on the 18th of April finally atlx fed as to boys, First of all, I agree we should hold up the publi cation of the chicken work until the current study is coapleted. .* * - the only other eoonent I hare of substance la that Z wish we could vo~k la several paragraphs reviewing the Troon work on vapors published la 1956 (see en closed xerox copy of reprint), k logical, place for a review of those data would be at the top of the ~ 3rd page of your review paper, before your paragraph at the top which begins with *Durlag succeeding years llttla Information was reported concesnlng . the toxloAlpgy of tneJC3j3r>til goLaugh! in etal, 1963. eta.- . * . . * *. ` . Host of the review articles on the PCB'a have failed to indicate awareness of the Treon work, Z believe this is because the Rlsebrougha, Peakslie, eta,, ete, -- the envlronaentallsta -- have not encountered the Antx'lcaa Industrial Hygiene Association Quarterly in the review of the literature nor .la It likely that . they ever heard of the AXHA, ^ Zn any case,' Z think the work la worthy of review even though it was inhalation data particularly since Drinker's earlier work in *37 1* frequently nentlooed and. this. . again concerned inhalation, ' . r- ' ;' * ? *.*i " ' , ' s- * .. . Zn the last paragraph of'this sane review paper you Qn(JC~Q y t \ SCH 023589 ^ WATER PCB-SD0000012623 Dr, Otis Fanchtr April 28, 1972 Paga - 2 - refer to tha parniaeabla aablent concentrations of PCB* a" aa suggested by Xlrinicer In 1939. Aotually the ACOXB established tha Threshold Halt Valuta baatd oo Trton'a work rathtr than the tarlltr Drinker work. Z btlltvt wt aay have arrived at our high dlttary level of 100 ppu in tht atudlta dona at Bio-Teat baatd on tht Treoo work rathar than Drlnkar'a. George Ztvlnakaaa la reviewing taeh of tht papers ' to aaa If you. Bap and I hava Biased typo'a and ha ay wall suggest soot graanatlcal changes igjoh wt will antar on our eopy and aand xerox's to you and Bap. . Z hopa to gat ooplaa of all of tha atudlta in tha hands of Bill Fapageorge, Scott Tuoktr and tha lawyers next wtak. Z do not antlolpata a lot of ohangta frou thouand hopa that tha attorneys agrta that wt oan go ahead with publication. Boat personal regards. Sincerely, m/bka cos M. L. BepUngtr Xlatr P. Wheeler Mgr., Xnvlronatntal Health Medical Dap&rtaant OnQCZl . SCM 023590 WATER PCB-SD0000012624 Xh i I V \ It B I T i WATER PCB-SD0000012625 * / HnduMd, BI O-TEST Jldyyuzt&ues. Snc. 1810 PRONTA3C AC AO roiieeiaOT CHvlAOMM CMfAV tOCNCCt CMCMltTIr ' PUAf SCJCNCCft MCOtCAV SClCNCCJ NOATHBROOK. ILLINOIS 80082 April 18, 1975 ArR 21 ACA C30C ju t(lc#monc j;mcjc Dr, George Roush, Jr. Mon seats Company 800 North Lindbergh Boulevard St. Louis, Missouri 63166 Dear George: - 1 fully appreciate that the meeting an PCB's today was not completely satisfactory and that many nagging Questions remain. The enclosed is a brief summary of my personal views and I would appreciate any open and frank comments that you all may have. Please let me know of any action that you contemplate in the way of seeking additional assistance in pathology or in contacting federal agencies. We will be pleased to be of help in any way that you may wish. It is my feeling that we need to get together again within the nest few weeks to continue our discussions. * Very truly yours. JCC:AR J. C. Calandra President cc - Dr. George Levinskas Mr. Elmer Wheeler Mr. William Papageorge SCM 022745 DEPOSITION i EXHIBIT jlEVHJSgfrS-H I *s -wn ^ '`QCSC WATER PCB-SD0000012626 ix m WATER PCB-SD0000012627 .<w 'd i^ .*- - . *ir .''**: *'*?*'* Dr* Ctorft UtUckai HwiMito 't`v . , ^~ . . * * =-.^y ^ c. -- - ` -' - * v**V ' - bii^iinltf rtT-^aa.4 u addWooal: ^ _ ,'jMtlw of Uwtltgg^Ui^KtjSjfl^^yp^^'^Srpqy TH f lOppm imli ;*:,; ' of 1BT tt.'T(ti^lH>^M'|mioit''ini Sit jtmn ibt original block f wibiddol'Uin*^;.Jk^M(ilc Itilo* ii wi|lllT claaalDod by Or. KMtorui hopelocal.' Tariaw/,:Pr. hiehtnr hu eluiUItd thla loaloo u nwhlu kyptijptttU *ad I ooacmr. Howvvar, you will ilio nolo that whan addlttooalMctiona of Uvor from thla aoimal war* pntiiHd (ST Study a* U*r alia rations war* ': wa(( which attoatsto tkiSial Man it thaso hilsaa^. ............................... ; ftport W 1BT / ,rw>- &toijy istUa oi tbi bm . m "" ` ' ' ' " " V. i v.f.- v. . _ V . ..;-- * - /. ft E. Gordo*, 0. V. M.. Pb. D. . Sactioo Hoad. Pathology Obptrtmit OOOCC3 WATER PCB-SD0000012628  1 P & 2. j::::::i:j:j::::J; +1>* OK D fr So Sr 1 : s :: j ::::;: j j tj : i j ;: j; t! I X* X * X t cn EA l*- 1o r? i: > J? s xt s.? Cytoplasmic vacuolatlon of hapalocytaa Focal oacroala of hapalocytaa Focal lymphoid Inflltratlooa Focal byportrophy of hapalocytaa o *o *d d O'd'd *d Nodular hyporptaata of hapalocytaa c Hi >> Focal blla duct kyporptaala r m Se' Hapaloma a *u o CliolanglO'hapaComa natlculum Coll Sarcoma motaatailc Mammary tumor, mala static T*` 1 t 1 **!.. f*M WATER PCB-SD0000012629 uixx I B I T # WATER PCB-SD0000012630 s Monsanto MoflltntS Comgiity 800 N. Lindbtrgft louUvtrd St. louii. Umouri 83108 Fton: '314! 194-1000 July 18, 1975 Dr. J.C. Calandra Industrial 310-TEST Laboratories 1310 Frontage Rd. Northbrook, 111. 60062 re: AROCLOR 2-year Rat Feeding Studies Dear Joe: The attached table summarizes a comparison of the 3 revised. AROCLOR reports (12^2, 125^, 1250). ' In 2 instances, the previous conclusion of "slightly tumorigenic" was changed to "does not appear to be carcinogenic". The latter phrase is preferable. May we request that the AROCLOR 125report be amended to say "does net appear to be carcinogenic'. The number of hepatomas reported for AROCLORS 1260 and 12^2 have been interchanged. This appears to have arisen from con fusion regarding the numbering of the animals. The original reports show tumors in animals with numbers in the 100-300 range for AROCLOR 1260 and in the 500 to 800 range for AROCLOR 12^2. This leads me to conclude that the numbering scheme shown in the second set of reports is correct. Vith AROCLOR 125** confusion is compounded. The original report showed tumors in animals with numbers in the units to teens, but the revised report shows animal numbers ranging from 40 to 1000. Can this be straightened out? I was unable to reconcile the differences in the animal numbers between the first supplemental report and the original reports, I had inquired as to the changes in the numbers. As I recall, I was told that the sections had been renumbered when the new slides were made and that a key relating to the sets of numbers received Zjui 1 l-o WATER PCB-SD0000012631 Dr. J.C. Calandra July 18, 1975 Page - 2 - would be supplied. This has not been done. It may not be necessary for AROCLORS 1260 and 1242, but AROCLOR 1254 remains unresolved. Insofar as I can see, the remainder of the reports appear acceptable. Kindest personal regards, Sincerely, George J. Levlnskas, PhD Mgr., Environmental Assessment and Toxicology /bkp att. cc: Dr. George Roush, Jr., M.D. QnQCCr r 04 4 m WATER PCB-SD0000012632 Product Supplemental Report #1 (mailed) AROCLOR 1260 . conclusion hepatoma* s. slightly tumorigenic t* t3 range of p. 9 p. 10 p. 11 p. 12 P. 13 p. 14 test animal nos: 600 to 800 1000 series 70 to 100 500 to 600 600 to 700 700 series Supplemental Report #2 (JCJ delivered) does not appear carcinogenic 7 100 to 300 800 to QOO lOtto 40 80 to 200 200 to 300 200 to 300 AROCLOR 1254 conclusion hepatomas slightly tumorigenic 6 slightly tumorigenic 6 . AROCLOR 1242 conclusion . . slightly tumorigenic does not appear carcinogenic hepatomas range of test animal noa. 7 3 as in report #2 as in report #1 for AROCLOR 1260 AROCLOR 1260 SCM 02280^ o^oc WATER PCB-SD0000012633 LUXI II I B I T # l I ! i Ii i i WATER PCB-SD0000012634 fouicocoar (nvhonmimtav MCI CM CM' *T*V LAMT ICKNCfl MCSICA4 ICKNCfl JfutluA&iuil BIO -TEST J&lxyuitiUA. 3nc. 1810 FRONTAGE ROAO NORTHBROOK. ILLINOIS 80062 August 4, 1975 AmtA eooc jit rtkCPMONc trt-iojo Dr. George J. Levinskas, Manager Environmental Assessment and Toxicology Monsanto Company 800 North Lindbergh Boulevard St. Lotus, Missouri 63166 Dear George: \ ' Re; Aroclor - 2 Year Rat Studies . " in regard to the comments and questions covered in your letter dated July 18, 1975, pertaining to the above, please note the following: 1. We will amend our statement in the last paragraph on. ' page 2 of the Aroclor 1254 report to read, "does not appear to be carcinogenic" in place of "slightly tumorigenic" as requested. 2. In regard to the animal numbers in the Aroclor 1242 and 1260 reports, they are correct in our final revised report. In the original reports, the Aroclor titles for these two materials were reversed. 3. The animal identification numbers appearing in the reports on evaluation of additional liver sections are the same as those in our original report. The animals were not renumbered. 4. We cannot find any discrepancy in animal identification numbers in the reports (original, re-evaluation, final revision) on Aroclor 1254. However, in the report on re-evaluation of additional liver sections dated March 24, 1975, there was a typo graphical error on page 1 which referred to Aroclor 1260 instead of 1254. Perhaps this is the basis of your confusion. I hope that this will serve to further clarify the situation. Thank you for your assistance and cooperation. JCC:AR Sincerely yours, J4*- J. C. Calandra President ,, POSITION" j], , exhibit fi^iiSSJSrA. L* ICl)A- SCM 023620 00QC.G3 WATER PCB-SD0000012635 w xx I v I I WATER PCB-SD0000012636 <' B. Summary la Dott iMtiBUt, the spectrum of tnilwitf'nUlid histopathologlcal findings la the liver from this rctviluattoa did nod cfllfcr significantly from that previously reported la our original report dated November 12. M71. There were six hepatomas detected among six of the artamis at the highest ' treatasnt level (100 ppm) of the 24-Month Sacrifice. No hepatocellular . '. . >--i carcinomas were observed. One UWr tumor (a hepatoma) previously reported , .in a T-tl. animal {Noiv^fl) vas re*clusified aa nodular hyperplasia. The *W ,v 'other reattarat-related lesion* reported are regarded as d` egene'rative or " hyperplaatlc in nature and th.yy ire morphologic feanlfeetitions of aa acapd'-e - *' ` ` 1 " .. response of the Uver ascribed to biotrans/ormation of the test material. The 1 Utter Uetons were confined primarily to teat anlmaU of the 12 and 24 month V.. sacrifices .. and1 they were doae-related In incidence and severity. , In conclusion, Arocler US4 does not appear, to bo carcinogenic in rate *. - ... = di; \i/ . c. j . fed lor two years at'leveU up to and including 100 ppm. \ i'-1' Respectfully submitted. v INDUSTRIAL 310-TEST LABORATORIES. INC. WATER PCB-SD0000012637 x x rti I V I l I1I WATER PCB-SD0000012638 \ > T : Xfe- . ... vrr: > Otv&iUtkusl IIO -TIST Jai&aIo*ieA. Ate. V 1*10 FRONTAOC ROAD NORTMiROOK, ILLINOIS MM3 /' August 5. 1975 172 George J. Levinskas, Ph.D. Monsanto Company 800 N. Lindbergh Blvd. St. Louis. Missouri 83165 ` Dear Mr. Levinskas; . Res 1BT No. 641-06672 - Hlstopathological Evaluation at Additional Liver Sections from Rata ot a Two - Year Chronic Oral Toxicity Study of Arcelor 1254 Enclosed please find 3 copies ol page 2 from our revised laboratory report dated March 24, 1975, prepared in connection with the above study. Very truly yours, . ^ . "1fa* ' J. C. Calandra ' - President JCC/fd S DEPOSITION J EXHIBIT ^Lci)iv<>fcAV <34 i ude r WATER PCB-SD0000012639 E X H I B I T WATER PCB-SD0000012640 August 14, 1975 Dr, J.C. Calandra Industrial BZ0-TS3T Laboratoriss 1810 Trontags Road ^ Sorthbrook, HI, 60062 rat AROCLOR 2-year Bat Feeding Studies Dear Joe: With respect to your 1 attar of August 4, 1975* we appear to be reading froa different aerlpta, . The attached coplaa of pages froa the revised AROCLOR 1254 report (ZBT So, 641-06672), dated March 24, 1975 shows -that virtually all anlaal numbers have 3-dlglts, Sts exceptions Include a'A-dlgit number on page 10, a series of 2-dlgit numbers ranging froa 71*70 on page 11, and slides marked Set or Zx'appearlng on pages 12 and l4. Copies of pages 83*88 froa the original report, Z3T Jib. B7298 dated Vovember 12, 1971 also are attached. These contain the tables listing tumors, and they are the only ones in which individual animal numbers appear. As summarised below, there is a repetition of low digit numbers in each group. Male ret nos. Female rat nos. Control 1,2 W-Wf50, AHMLfiM I2b4 1 1 ppm 10 ppm 100 ppm 1*2 3-13 1.2 3-H 4,5.7 1-3.6* 8-15 The only pages which contain similar numbers are page 11 of the revised report and page 83 of the original report. How- 2 DEPOSITION 1 i EXHIBIT 1 ^>1-21 U*C I SC" 22901 Q000G1 WATER PCB-SD0000012641 Dr. J.C. Calandra August 14, 1975 Page - 2 - aver, even in those Instances share the saaa numbers appear, there are inconsistencies as shown below1 animal no. 46 l a4s 50 52 12 66 if,roup and sex 83 1 ppa female 1 ppa female 1 ppa female 1 ppa female 10 ppa Male 10 ppm female 100 ppa Male 100 ppa female all shown as control females Although "it is of minimal significants, pages 84 and 85 of the original report are aisnumbersd in that page 64 is a continuation of the table which begins on page 85. The page for the AROCLOR 1254 containing the phrase "does not appear_to be carcinogenic" has been received, Ve note the interchange of the numbers 1242 and 1260 on those 2 AROCLOR reports, and will destroy the nlslabelad copies. Sincerely, /bkp att. George J. Levinskas, IfcD Kgr., Xnvironaenta 1 Assessment and Toxicology 02zaoa sc* 000CG2 WATER PCB-SD0000012642 WATER PCB-SD0000012643 Toneouoar ' CNVlAONM CMTAk ICIfNCtl CMCMIITKT ALAHT SC1KHCCS . MCOICAI ftCICNCCS 9M&jbt*tAjjal BIO - TEST JPd&uiiyu&. 9nc. 1810 PRONTAG8 ROAO NORTHBROOK. ILLINOIS 60062 October 17, 1975 -Ffrf f'J* aca cee* nt mtfHSMC 171-lOJO George J, Levinskas, Ph. D. Mgr., Environmental Asaesament fc Toxicology Monsanto Company 800 N. Lindbergh Blvd. St. Louis, Missouri 63166 Dear Dr. Levinskas: I am returning the black and white photomicrographs of Aroclor lesions in the rat liver that were taken by Dr. Pour at the Eppley Institute in Omaha, Nebraska and delivered to me by * Jim Hill. I found his report interesting, although I do not concur with his classification and interpretation of some of the liver lesions. Per your request, I am also enclosing a copy of Dr. Kimbrough's findings and report on Aroclor 1260 in the rat which you sent to me earlier in the year. Sincerely, DEG/ji enclosures Donovan E. Gordon, D. V. M. , Ph. D. Section Head, Pathology 3 deposition } EXHIBIT |tgy<U5K/ta-<3k 00QCG3 WATER PCB-SD0000012644 U JX X \ I B l T # I 1 ii WATER_PCB-SDOOOO012645 ii s, ** CUC LdeO INTER-OFFICE CORRESPONDENCE 1IO-TIST /timmktrn Am. Mlt DaCMiUr 24, 1175 u*jcr. PCS Paper* 003 George Lrvinskas haa lome questions regarding the nathology section of the PCB papers. He want* to visit Northbrook alter Jan. 1 at a time convenient to you. Please make the necessary arrangements. CLK/lam G. L Kennedy s. WATER PCB-SD0000012646 IU X X ( v I B I T # J WATER PCB-SD0000012647 Dr. U*ioift'wttfbi ltm -... A&***> *?'%*$; th pttholo|| Mellon lb PCD P4*? fl* ^^1 ^^' /r > . - . *... .'.'-V%s c ~*^y.;V*.1-iv ** ! r\*~V, v*"- '.*?5- ** *iC rl ' l* #V 1a^L v * * r*~ -'. .. ,,;.* ' " * * \ . " . r '' " "*: *-. ; ` . ..... . , K, January 11. m ' , .;/ V -*y : "V ' ` * l'*i ' . ** * .> _*^ * **' m 1 T ' ' ' Dr. Livlniliti lui roachodultd hit trip for Thursday. January IS. 1 *. </. ffiii'.'.-000GG5 WATER PCB-SD0000012648 UJXX I B I T # ( t\ WATER_PCB-SDOOOO012649 ) Toxicity and Environmental Effects of Commercial PC] Introduction . Polychlorinated biphenyls (PCBs) are not a single chemical entity. They are produced by chlorinating biphenyl to acheive a final product with specified properties. These products, sold under the trade name AROCLOR by Monsanto, are mixtures of chlorine-containing biphenyls with different numbers and positions of the chlorine substituents. Over the years, a series of investigations have been conducted to assess potential health and environmental hazards of PCBs. The 3 predominant commercial mixtures (AROCLORS 1242, 1254 and 1260) have been studied most intensively. Toxicity' tests performed on these 3 commercial mixtures were typical in scope of those designed for the evaluation and establishment of the safety of direct and indirect food additives. It may be noted that AROCLOR 1260 no longer is produced in this country since it does not meet the physical specifications for its restricted uses. Conclusions Based on Monsanto Studies 1) As a class, the AROCLORS are relatively harmless materials for routine industrial handling under ambient conditions. 2) Threshold Limit Values of 1 mg/m^ and 0.5 mg/m^ have been established for materials containing average chlorine values of 42# and 54#, respectively, of the available sites. 3) Tfre no-effect level for these materials in chronic 2 DEPOSITION ( EXHIBIT 1 S-ai-81 toic SCH 019639 flionr.i jo WATER PCB-SD0000012650 .0 ) rat and dog feeding studies is about 10 ppm in the diet. No hepatocellular carcinomas were present. 4) The no-effect level on rat reproduction is between 1 and 10 ppm in the diet since higher levels result in - low mating indices. 5) No teratogenic or mutagenic effects were observed in studies with rats and mice. 6) In chicken reproduction and teratology studies, AROCLOR 1242, which produced the most severe effects, had a no-effect level of 2-4 ppm in the diet. 7) Acute toxicity to fish Varies from less than 1 ppm to greater than 100 ppm depending on the specific AROCLOR and species of fish tested. 8) PCBs containing 3 or less chlorine substituents per molecule are reasonably biodegradable. Summary of Monsanto's Long-term Toxicity Studies on Commercial PCBs. These tests consisted of 2-year (lifetime) feeding to rats, 2-year feeding to dogs, 3-generation rat reproduction studies with 2 litters cast per generation, rat teratology studies and dominant lethal mutagenic studies in mice. Toxicity and reproduction studies in chickens were performed to evaluate possible untoward effects In birds such as decreases in eggshell thickness. In addition, biodegradation and tissue accumulation studies have been conducted. The highest dietary level (100 ppm) in the lifetime rat feeding studies produced a weight depression at 24 months in females fed AROCLOR 1254 and liver weight increases in all SCM 019640 000CG7 WATER PCB-SD0000012651 / groups except males fed AROCLOR 1242. As with other halogenated hydrocarbons, the important histopathologic changes were present in the livers of animals sacrificed at the end of the study. These consisted of hepatocellular alterations sdch as focal hypertrophy, cytoplasmic lipid changes and in some animals from the 100 ppm groups, hepa tomas or cholangiohepatomas. No evidence of hepatocellular carcinogenicity of the AR0CL0RS was found. In the dog 2-year studies, some slight depresses in body weight gain were noted. At the highest feeding level (100 ppm), AROCLOR 1260 produced an increase in serum alkaline phosphatase activity and liver weights without concomitant - histopathologic changes. However, there was evidence of gastrointestinal inflammatory lesions and ulcerations which appear to be similar to those found by Allen in rhesus monkeys fed AROCLOR 1248. (AROCLOR 1248 was an experimental product which never was commercialized.) In the rat reproduction studies, none of the AR0CL0RS produced adverse effects in the 2 litters of the first gen eration.' In the second and third generations, there was a reduction in the mating index at the 2 highest feeding levels (10 ppm and 100 ppm). The ability of females to conceive, carry the delivery process to parturition, and to successfully nourish the young was not affected by the 3 AROCLORS. No changes were produced in the reproductive tracts of either male or female rats by any of the 3 AROCLORS. There was no evidence of teratogenic or mutagenic changes with any of the PCBs at maximally tolerated dose levels in SCM 019641 00QCG3 WATER PCB-SD0000012652 u) rats and mice. In the chicken toxlclty/reproduction study, AROCLOP. * 1260 was without effect at all test levels. AROCLORS 1242 and 1254 at 100 ppm In the diet decreased egg hatchablllty. In fact, poor hatchablllty of eggs was found from hens fed 8 ppm of AROCLOR 1242. In addition, AROCLOR 1242 at 10 ' and 100 ppm and AROCLOR 1254 at 100 ppm were associated with reduced eggshell thickness. Chick viability was affected by both substances at a-dietary level of 10 ppm. Biodegradation studies were conducted using semi-contlnuous activated sludge systems and tissues of rats fed PCBs were analyzed to measure accumulation and retention of these ma terials. These factors are influenced by the number and ` position of the chlorine substituents. Higher chlorine-con taining members are more resistant to biodegradation and they accumulate more readily and are less readily metabolized and/or excreted from lipoid tissue in animals. PCBs containing 3 or less chlorine substituents per molecule are reasonably biode gradable. Comments The conclusion that "No evidence of hepatocellular car cinogenicity of the AROCLORS was found" in the 2-year rat feeding study was reached only after extensive re-evaluation of the original liver slides as well as additional liver sections from all of the animals after the Kimbrough results on AROCLOR 1260 became known to us. The slides were read SCH 0196<2 ooocco WATER PCB-SD0000012653 (J f independently by Dr. D. Gordon of Industrial BIO-TEST Labora tories, Professor W. Richter of the University of Chicago, and by Professor P. Pour of the Eppley Institute for Research in Cancer. In addition. Dr. Pour evaluated the Kimbrough slides and does not agree with the reported findings. Barsotti and Allen have reported that 2.5 ppm of AROCLOR 1248 in the diet of rhesus monkeys adversely affected repro duction. This approximates a dosage of 100 pg/kg/day. It is higher than the safe human intake (1 pgAg/day) estimated by PDA from human data when it promulgated its tolerances for PCBs in food. OX SCH 019643 onor.70 WATER PCB-SD0000012654 o) . . Human data (F.R. July 6, 1973. p. 18097, Section (2)) Lowest level of PCBs producing effect In man 500 mg total. 500 mg consumed by 50 kg Individual over 50 days. . 500 mg 4- 50 kg 4. 50 days 200 ug/kg/day (effect level) _ Using 10-fold safety factor leads to estimate that 20 pg/kg/ day may be-safe over a 50-day interval. Since PCBs have long half-life; calculating same Intake over a 22-month span arrives at an estimated safe intake for a pro tracted period of 1 ugAg/day. Primate data (Ped. Proc. 2^:338 (1975). Abstract 675. Barsotti & Allen) At 2.5 ppm in diet, primates Ingested 182 mg over 52 weeks. 182 mg n 365 days 0,5 mg/day which produced effects. Assuming 5 kg primate, 500 pg 4- 5 kg = 100 ug/kg/day Conclusions: Primate study done at dosages (100 ug/kg/day) greater than the safe dosage (1 ugAg/day) eslmated from human data. Primate study does show effects at lower dosages over longer time span (100 ugAg/365 days) than had been observed in man ' (200 ugAg/50 days). The tooal amount of PCBs producing effects in primates (18^ mg) was lower than that producing effects in man (500 mg). (N.B. Abrahamson & Allen, Env. Health Perspectives. June, 1973, bl-66. Report that infant monkeys are able to tolerate doses of PCBs that produce extreme morbidity in adult monkeys.) Dog and Rodent Data (F.R. July 6, 1973. p. 18097, Section (1)) nNo-effect" level for man using a 100-fold safety factor and accepting 10 ppm as a "no-effect" level in animals would be 2.5 ugAg/day based on dog data and 3 ug/kg/day based on rat '' data. \ ' ' '' .............-- If rat data "no-effect" level drops to 1 ppm, then corres ponding estimate of human "no-effect" level drops to 0.3 ug/ kg/day. Question: In a "collision" between rat data using a 100-fold safety factor and human data using a 10-fold safety factor, which data base would you like to be riding cn? Comment: Since human data is available, it could be argued 'that the traditional 100-fold safety factor is not necessary. Application of a 30-fold safety factor to rat data, supported by dog and human data, makes present estimate of safe human intake appear reasonable. SCM 0196<<r 000071 WATER PCB-SD0000012655 1 ) f PCB Primate and Human Residue Data Cone, In diet, ppm Intake, ug/kg/day Estimated safe dose, UgAg/day Liver eone.,ug/g Eat cone., ug/g Primates 2.5 100 Human 56.3(0.01)* 50,0(27.7)* 0 |3l4 samples] ** <1 125 samples 1-2 ]l65 samples] >2 [33 samples) Value in ( ) from Infant primate. Allen et al., TAP 0: 10-451 (1974) **Yobs, Env. Health Persp.,79-81, April 1972 SCH OHO"'5 000 \ t 4 *+ WATER PCB-SD0000012656 E X H I B I T / WATER PCB-SD0000012657 ncnvMT OCtaCCB CtC*<(* MCOiCAt leukcct *-; ' NOtTHtllOOS^ULlNOliTJOfl*^^ SfiS&Sg;>/*'' .f *-V .17 .. , .' ; ? .-.pg*. ,)Wj^^V^rpt "r/f ............. %' Prji, ,'ii;\- f.ts1 . /.'it.?'?,,#:...Tttgffiry::.f .*'' .,,y 'i '.' >$>>. * ^X3gl^-v*:iL_'3B>r : *% :,?>. *. " # - ' * C<-*orpe J. Lpvinikua. ih. D. Mon*.into Company _ 800 N. Linil>*rpl lUvil. '"' * * ' . ** V s'*s ;va**..- t 6 T^O` P^. i . .|' >* ' &,# - St. Louis, Mi.vourl 6il66 - '. Dear Dr. Uvimlui*: /Jf/VV v-& I am enclosing the rcvieotThiriwr tafetes /er the Aroelor p*p*r that Ut-o diacudKi'd during your \isit on Jawwry 15* 19?^ Jfou will ., - _ notice the re arc lw laities with the satire data* * Oae^tabte la a listing of tumors by organ system, as you suggested* lo repUce' tbc urtsliul hble, I think I also prefer the former although it will require inoro space when printed. 1'' - >.*. , 1 .*. - ..-^v: ' - ; \ ^ j* f. J*' . ' Should you haver any further qwilionr, please contact me* '* A - '.*'' VT'" & . ` -* ' ^ ` '"*' * ** v - ^ * Sincerely yours, --\ ' ' ; * ,^> -- i . -...a* v A _':. - . r % - " r# #-/"/ : * **'' , * D*nnvnn B. Gord**n*` DV.M* Ph. D* - Sot?turn Ifuid l^thulottV ' DBG/ji : . cc: M L. Kcpllnpur.^ ^ ;'*v .V M : .& 1 DEPOSITION 1 } EXHIBIT I |U[Vl05fcflS-3c| 1 S'3H'2l ^c| ortnr*7'T WATER_PCB-SD0000012658 E X H I B I T f WATER PCB-SD0000012659 m Monsanto Dipt. *>f Medicine & Environmental Health** / * ' Levinskas, A2SA H_ i f* November 17, 1978 AROCLOR 1260 TO FILE i ' G. Roush, Jr. - A2SA J.C. Weber - B2SK G.F. Barton - A3NC W.J. McCarville - A3NC C.F. Callis - B2SA J.T. Garrett - A2SA ~ DEPOSITION J EXHIBIT f Lgv (fc&EftS* 51 On Wednesday, November 15, 1978, I had a call from Joan J U34L. Ranson, an industrial hygienist with OSHA in Philadelphil?^"^"TM^"^^ About two weeks ago, a Philadelphia trucking firm picked up two used transformers Westinghouse was having shipped. The truck carried other cargo of a consumer nature, namely shoes, toys and candy. The transformers were not accepted at the receiving end because their contents had spilled. Consequently, the driver took them to the truck terminal. The transformers contained AROCLOR 1260. An estimated 168 gallons of transformer fluid was spilled. CPSC has been tracking down the consumer products. Most are believed to have been recovered. EPA is on the scene. Details as to exactly what they are doing were not discussed at length. They apparently are offering advice on cleanup of the spill. Two local TV stations have camera crews at hand, and the emotional level is rising. The two leaking transformers have been removed to an enclosed area in the truck terminal. Ms. Ransom wanted to know about monitoring air levels in the enclosed area for PCBs. She also inquired about clinical or other tests which could be done on exposed workers to gauge their exposure and the nature of protective equipment which clean-up crew members should wear. SCM 019637 Without reiterating all aspects of this somewhat lengthy discussion, I made several statements to Ms. Ranson. She was told that there were no tests we know of to measure the after effects of exposure, and none should be necessary since there was no reason to expect that persons exposed under the conditions she had described would become ill from PCBs. With respect to air monitoring, the analysts should be careful to identify PCBs as such since AROCLOR 1260 was mixed with another chlorinated hydrocarbon. The latter was more volatile and, if detected, and expressed as PCBs, could create unnecessary concern in addition to being misleading. I saw no reason for concern about the health of employees' families from possible contact with PCBcontaminated workclothes. With respect to the emotional tensions, I suggested that they tell people there was no health hazard involved, but that efforts were being made to limit the potential environmental effects of PCBs. The use of disposable or protective clothing was justified to keep workers from taking soiled garments home and having PCBs flushed down the drain. THere was no need for respiratory protection for cleanup crews. Finally, if the truck had a wooden floor-bed, it IN-10M IREV. 3/78) OOOC74 WATER PCB-SD0000012660 FILE - AROCLOR 1260 November 17, 1978 Page - 2 - ) might be advisable to replace it. The loading dock apparently is concrete. All wastes, including spillage initially swept up with sawdust, should be disposed of in an acceptable manner according to prevailing regulations. Ms. Ranson appeared to be aware of those requirements. I don't know how many places have been contacted. Ms. Ranson stated that they had called Dow in the mistaken belief that they manu factured PCBs. They received a suggestion that adipose tissue assays might provide a useful measure of exposure. My comments on that suggestion ended with the observation that obtaining fat samples by biopsy would be the most traumatic part of the entire episode for the individuals involved. Ms. Ranson expressed appreciation for my remarks and indicated that her supervisor, Mark Durham, Sr. Industrial Hygiene Supervisor probably would call me tomorrow - Thursday, November 16, 1978. . Let's see what happens. . /bks Ge _ skas SCM 019638 WATER PCB-SD0000012661 V \ E X H I B I T # WATER PCB-SD0000012662 Monsanto noH in*m( ft tociriow, G. J. Levinskas - A2SC September 25, 1975 PCBs TO G. Roush, Jr. A2SA H. S. Bergen - B2SL D. R. Bishop - BIND W. B. Papageorge - B2SK R. G. Potter - B3SA W.W. Withers - B2SA The attached represents a final (?) version of the toxicity statement on PCBs. This takes note of all the comments which have been received since the version mailed to you on August 29, 1975. This was discussed on the phone with Wayne Withers, and he agreed that it was acceptable. Since Wayne was the only one who had comments regarding the August 29 version, this should be satisfactory to all concerned. /bkp att. Geo w . Levinskas SCM 019716 000070 WATER PCB-SD0000012663 PCBs Recently, we were informed that liver carcinomas were observed in female Sherman strain rats fed AROCLOR 1260 for 20 months. This prompted us to re-examine livers from male and female rats of the Charles River strain which had been fed AROCLOR 1242, AROCLOR 1254 or AROCLOR 1260 for 2 years in earlier studies conducted for Monsanto Company. There are 4 elements which are closely interwoven in this matter: (1) differences in test procedures, (2) differences in results obtained by various investigators, (3) definition of what is a cancer, and (4) evaluation of potential risks, if any, to man. These will be summarized briefly. - (1) Several animal studies have been conducted with various brands c-f' PCBs. In some studies, the test material has Been identified by trade name (AROCLOR, KANECLOR). In others, there was Just a general reference to PCB. Consequently, the quality of test material with respect to the amounts and nature of contaminating impurities or by-products cannot be determined in case. In addition, several strains of test animals were used, the duration of the experimental periods varied, and there was a wide range in the depth of detail with which the observa tions wei^e reported. Consequently, it is difficult to make comparisons between these studies. . (.2) .In general, studies have shown mice to be more susceptible than rats and females to be more sensitive than males to the liver effects of PCBs. Beyond these generalizations, the results have not been consistent. Some investigators have -- . SCM 019717 ooor.'?? WATER PCB-SD0000012664 reported liver carcinomas. Others have observed only- benign tumors. Several have noted changes in liver tissue without detecting tumor formation. For the reasons Just cited, it is difficult to determine the bases for these different results. The most direct comparison can be made between the 2 studies on AROCLOR 1260 since the same high dosage level of the same lot of AROCLOR 1260 was employed in both experiments. In the studies reported to us, liver carcinomas occurred in approximately 8# of female rats of the Sherman strain (the only sex used). In Monsanto's study, none of the liver lesions had progressed beyond the stage of benign tumors (hepatomas) despite a slightly longer duration of feeding of AROCLOR 1260 to rats of Sprague Dawley, Charles River strain. The Monsanto study employed rats of both sex and it did confirm the previously noted greater sensitivity of female rats'to liver effects of PCBs. (3) A review of the liver alterations seen in all 3 AROCLORS in Monsanto's studies shows that the lesions were benign in character in the traditional pathologic sense. An evaluation of all Information available to us, including the contradiction . between the recent data showing AROCLOH 1260 to be a carcinogen and our earlier negative results on the same product, leads to a conclusion that AROCLOR 1260 is not carcinogenic to all commonly used strains of laboratory test animals. `~ (4) In 1972, the manufacture of AROCLOR 1260 was dis continued in the United States and the sale of AROCLORs was restricted to a single use, i.e., as dielectric fluids. As such they are used in closed systems which will at least mini mize additional environmental contamination. Considering the ' SCM 019716 " 00007a WATER PCB-SD0000012665 high degree of fire risk associated with this use, and recognizing that AROCLOR 1260 may have a- weak carcinogenic potency which has not been fully proven and that AROCLORS 1242 and 1254 have not been shown to be carcinogens .Jit is _____:---------------- ------------------------------------------------------- ttriZsT concluded that the continued use of AROCLORS^as dielectric fluids will not present an unreasonable human health hazard SCM 019719 GOOC/79 WATER PCB-SD0000012666 * bee V B. Paoageorge B2SX d U/i January U, 1979 Dr. Donovan B* Gordon Industrial BXO-TBST Laboratories, las* 1810 Frontsg* Boed Hbrthbrook, Illinois 6006* Bat Folychlorinstsd Uphsnyl (FC1) Dear Doni )''- . . ^^ / * . Inclosed are copies of three rsprints dealing *ith llvsr tuaorigsnliis of FOB'S* These sro front . (1) qann. (1972).1* 805* (2) Cann. (1973).il 10J-106. - (3) J. Bat'l Cane. Inst*. (1973).5!l637-l6*$. ' Tbs studios vers psrforasd with Jsponsss ostsrlsls (Canechlors), but^jhs a--outs aro of direct Intorsat to our seating oq^AROCLCR 1260,2 Ton and Dr. Blehtar should b faeilisr vrta-tns contents of thoss papers as you review the AROCLGR slides* Our nesting Is sehsdulsd for January 31 1975 In Roou 201, Building 37, at BCZ la Bethasda* Vs *111 be nesting *lth Drs* dab rough and 8qulrss* Bo apse Iflet Ins has bean set, but us ean plan on starting In tha noralng. I *111 con firm a apse ifIc tins and *111 ehsek *lth you or Cap on s place to stay the night before* Sincerely, nl* ee Dr. M. L. Ceplinger Oeorge J. Levlnekae, Ph. D. Manager, environmental Aesese- sent and Toxicology 2 DEPOSITION 1 EXHIBIT SCH 022820 OOQC >0 WATER PCB-SD0000012667 VE X H I B I T # WATER PCB-SD0000012668 Monsanto *M tuajfcf 1 '. - TO t Bishop - BIND U i "irVzl/x4W. Nov. 17, 1975 '.'Mr. W. R. Corey - JS? 'Mr. F. J. Fitzgerald - B2SA 2 Dr. J. F. Mietire - T2F ^3 7 'Mr. D. Wood - B2SD^(ri > 07 13- -'Mr. T. H. Bottini - B2CA J Bi. P.-OT'DeGcUiuu B3TIA ^Dr. W. C. Hammann - B3SA*''"' 'Mr. E. H. Harbison - DlK *,9t Dr G J./'Mr. W. B. Papagaorga-- B2-SK y. ^Or. C. Paton - B2SC j5'j v .Mr, R. G. Potter - B2SB Dry Q. Roush A2DA ^Mr. J. C. Weber - BIND J'1 7 ^Mr. W. W. Withers - B2SA-2''3'' gg.'T. HE.-Wright A2SC Attached is the second draft of a news release summarizing Dr. Pour's re-evaluation of Dr. Kimbrough's study. It incorporates comments- from Dr. Levinskas, Messrs. Papageorge and Wood. Please let me have your comments and/or approvals as soon as possible tomorrow morning. We need to clear the final version with Dr. Pour and have copies printed for distribution in Chicago on Wednesday, following Dr. Kimbrough's presentation.' I'll have to print the news release late tomorrow morning. Yovv cooperation is appreciated. M- A 3^^/^ /mks attachment D. R. Bishop 'Xfrt-.J&MA- Tt^v^Lcf IN 10 *CV. ` DEPOSITION I exhibit <-gV(N ICficSri `S'Z-'l-Xl oJf SCM O58058 goocgo WATER PCB-SD0000012669 xxm I V / * WATER PCB-SD0000012670 Monsanto MOM lUiMI * V o*ri . D. R. Bishop - BIND Nov. 17, 1975 t <*ct TO : Mr. T. H. Bottini - B2CA Dr. P. 0. DeGarmo - B3NA Dr. W. C. Hammann - B3SA Mr. E. H. Harbison - DlK Dr. G. J. Levinskas - A2SC Mr. W. B. Papageorge - B2SK "Mr. W. R. Corey - B2SA Mr. F. J. Fitzgerald - B2SA Dr. J. P. Mieure - T2F Mr. D. Wood - B2SD Dr. C. Paton - B2SC Mr. R. G. Potter - B2SB Dr. G. Roush - A2SA Mr. J. C. Weber - BIND Mr. W. W. Withers - B2SA Dr. P. L. Wright - A2SC Attached is the second draft of a news release summarizing Dr. Pour's re-evaluation of Dr. Kimbrough's study. It incorporates comments- from Dr. Levinskas, Messrs. Papageorge and Wood. Please let me have your comments and/or approvals as soon as possible tomorrow morning. . We need to clear the final version with Dr. Pour and have copies printed for distribution in Chicago on Wednesday, following Dr. Kimbrough's presentation. I'll have to print the news release late tomorrow morning. Your cooperation is appreciated. ' /Q f\. /mks attachment D. R. Bishop a IO fcl'. 1 - . SCM 058337 cnoooi WATER PCB-SD0000012671 : IMMEDIATELY 1975 . _ " MONSANTO INDUSTRIAL CHEMICALS CO. D. R. Bishop (314) 694-2891 PUBLIC RELATIONS OEPARTMENT 800 N. Lindbergh Boulevard SL Louie, Missouri <3166 CHICAGO, Nov. 19 -- Monsanto Company today announced that a newly completed scientific report, commissioned by the St. Louis-based company, does not confirm the presence of malignant liver tumors in experimental rats fed a commercial polychlorinated biphenyl (FCB) mixture...a conclusion that had been previously reached and widely reported by Dr. Renate D. Kimbrough of the Center for Disease Control of the U.S. Public Health Service in Atlanta. The Monsanto-sponsored report, a re-evaluation of Dr. Kimbrough's work, is titled, "Histopathological Re-evaluation of Tissues from Sherman Rats Fed Aroclor 1260," and is authored by Dr. Peter Pour, a pathologist with the renowned Eppley Institute for Research in Cancer of the University of Nebraska Medical Center . at Omaha. Aroclor is a Monsanto trademark for a class of chlorinated hydrocarbon Industrial chemicals it manufactures. In her study. Dr. Kimbrough reported that when Sherman strain female rats were fed 100 parts per million of PCB (Aroclor 1260) for about 21 months, 170 of the 184 experimental animals developed neoplastic lesions (precursors of malignant tumors). She further identified 26 of these lesions as hepatocellular carcinomas (malignant liver tumors) . SCM 058338 onor.32 WATER PCB-SD0000012672 --2 During his re-evaluation, Dr. Pour saw the same alteration but identified them as being hyperplastic or non-tumorous lesions, further pointing out that it was his strong impression that many of the detectable alterations were a degenerative and reparative rather than a neoplastic process. Dr. Pour's re-evaluation is consistent with conclusions drawn from Monsanto's own PCB feeding studies which have never produced a carcinogenic response in experimental animals. . In his discussion, Dr. Pour reported that he observed 20 cases of abnormal lesions which presented such structural criteria as to be considered possible precursors to tumors, although a most significant criteria for malignancy -- namely invasiveness -- was missing, and the sign of ongoing toxic, degenerative and regenerative processes in the rest of the tissue was evident. "At present, the statement that these lesions may have metastasized (infiltrated adjacent tissue) if the treatment had been continued is as much as unreliable as the possibility that they might have been regressed if the animal would have been kept longer or for life," he stated. "In summary," the Eppley Institute pathologist reported, "it is difficult at present to conclude whether or not some of the examined lesions represent a malignant lesion, because of the lack of invasion and metastases. In the case of such organs, as the liver, with its marked tendency toward regeneration, it is, in my -more- SCM 058339 0^0C03 WATER PCB-SD0000012673 --3 opinion, difficult and sometimes impossible to distinguish between regeneration, hyperplasia and neoplasia, particularly when the tissue is continuously exposed tp a toxic substance." Quoting further from Dr. Pour's report, he wrote, "in this context. Dr. Kimbrough's study seems of particular interest, in that polychlorinated compounds may be retained in the body, even in higher concentrations for several months after feeding has been stopped. This finding may explain the continuing degenerative and regenerative processes in the livers of most experimental rats, even two months after Aroclor was removed from the diet. "One should also bear in mind that some animal strains may react more specifically to a compound because of common endogenous diseases, as in the case of the Sherman rats used in this experiment which tend toward spontaneous liver lesions. "In my opinion," he concluded, "many of the lesions induced were of a degenerative nature, and without further studies, the induced liver lesions could not be definitely designated as neoplasia (tumorous). -0O0- NCTE TO EDITORS: Copies of Dr. Pour's report are available on request, in writing, from Public Relations Department, Monsanto Industrial Chemicals Co., 800 N. Lindbergh Blvd., St. Louis, Mo. 631 SCM 0583^0 GOOCG* WATER PCB-SD0000012674 U JX I V \ I B I T i 0 WATER PCB-SD0000012675 Sii/SjuAtAJ/il BIO -TEST J!&'h&vsJjyiuzb, Qrx. toxicology CHiMiim M CO I CAL SCICNCES MicAOciouoar N0U4T4IAL fAXCTT 4 HCALT* 1810 FRONTAGE ROAO ' NORTH8ROOK,ILLINOIS 80082 J\lly31, 1981 AAA COOC Jll TCLCAWONC *7*-1020 TCLXX 71-4417 Or. George Levinskas Monsanto Company 800 North Lindbergh Boulevard St. Louie, Missouri 63166 Dear Or. Levinskas: As per your request of July 30, 1981, Or. O. E. Gordon is attempting to locate the raw data for the pathology reports on histologic examination of additional liver sections from rats fed Aroclors 1242, 1254 and 1260 (IBT study no. 641-06672). These data will be microfilmed and a diazo copy sent to Mr. A. TJelner, Manager, Quality Assurance. Xerox copies of these data will be mailed to your attention. If I can be of further help, please contact me. Sincerely, MABjAR cc - Mr. A. Uelner Monsanto Company Marilyn A. Biederer Validation Assistance Specialist I- DEPOSITION { exhibit 5^nui SCM 023698 G'locrjs WATER PCB-SD0000012676 LUXX I B I T # 3(o WATER PCB-SD0000012677 Monsanto De, Jf Medicine & Environmental Health J. Levinskas, G2WF (4-83C9 July 9, 1981 AROCLOR Products 1242, Two Year Rat Feeding St a!nd 1260: J.R.G. Ortiz, E2ND A.F. Uelner, G2WC J.H. Craddock A complete audit of these three past IBT studies will require considerable time. So much time, in fact, that the results of the audit would not be available until long after completion of the monograph which is being prepared. As a result, it has been decided to limit the audit to a determination of how long animals were on test and as to whether or not liver reactions were taken for microscopy. These are the crucial elements for assessing potential carcinogenicity of these materials. GJL/mcl INOOM (REV. 3/731 SCM 058929 WATER PCB-SD0000012678 UJXX I B I T l WATER_PCB-SD0000012679 Monsanto C- Njme ol Nominee Paul L, Comojny Un.C at SUM 00l. Wright Oivition 8utmu Croud Oaoartmant location Creve Cceuj Corporate Staff Volition Tid* Medical Salary Grada Annual Salary Medical Oata of Lit incraait Toxicology Manage: Par'ormanca 19 $28,980.00 Growth 2/75 Data UK Emoioya Atvaw Excellent Excellent 2/75 Type of Award (Select tfta award category from tne opposite tide of tn>i ineel wfln most eppfopeietety oescriots me award and antar tna category mo __ code oalow.) Staff_____________________________________________________________________________________ Award Category Category Code Numoa Solution to product toxicity problem which permitted Monsanto 1 28 o<<rib in* icnivmam *nd its mniiicme* to Monunto oiowto continue product manuf acturing and sale. Dr. Paul L. Wright has performed his regular duties exceedingly well and has, in addition, completed a specific "ad hoc" assignment with dispatch and distinction. Dow informed us that they had encountered lung tumors in rats fed maleic anhydride and that they felt they had to report these findings to the Pood and Drug Administration (FDA) ait once under their "product stewardship" program. Dr. Wright was asked to contact Dow's toxicologists for details of their findings. This he did, and by his personal and professional! efforts he convinced Dow personnel that while they had an obligation to report their findings to FDA, it would be foolhardy to act precipitously. Instead, Dr. Wright advised Dow to defer contacting FDA until they had thoroughly evaluated their findings and a subsequent course of action had ibeen developed to allow resolution of questions which undoubtedly would be raised by their report of tumors. Dow agreed to this. Dr. Wright then contacted the Process Chemicals business group and informed them of Dow's findings. He proposed a dual course of action. The first : step was a commitment to support a repeat of the feeding study to determine whether Dow's findings were reproducible. The results of such a study would not necessarily resolve FDA's concern over the potential carcinogencity of maleic anhydride, but the intent to conduct the study promptly would serve to forestall precipitous action against the product by FDA. The subsequent step was to provide assurance to FDA that a thorough investigation of the toxicity of maleic anhydride would be undertaken, either ay Monsanto alone or in cooperation with others. This proposal was acceptable to the business group, and Dr. Wright so informed Dow. Subsequently, Dr. Wright accompanied Dow personnel to FDA when they presented their findings on maleic anhydride. FDA's toxicologists reviewed the data and the proposed plans of actions. They concluded that while the questions of carcinogenicity would need to be resolved, there was to apparent need for immediate prohibition of maleic anhydride or polymers tontaining maleic anhydride in food contact applications. Thus, we aelieve that Dr. Wright played a singular, outstanding role in preventing TDA from publicly proclaiming that maleic anhydride was a suspect Racommanded 6yt Oatd Award Amount Racommandadi Acorand 8yi O.i. Award amount aoorovcd lor payment'* ^ o-ijjj /t-r <?s- * T * 'JO / 2 DEPOSITION i. | EXHIBIT I ^7-31 y$2,500.00 k- SCH 058799 000:03 WATER PCB-SD0000012680 er." A ward Z i~. a t. . e 1 carcinogen. Because of the many uses of maleic anhydride in feed contact polymers, this was a significant accomplishment. Subsequently, when maleic anhydride was included in the priority list for study by CUT, Dr. Wright undertook writing of a literature review on the toxicity of this material. Notwith standing the fact that he had assistance in locating pertinent articles in the literature. Dr. Wright had to read, digest, and write a substantive review in a relatively short time. This meant considerable extra work on nights and weekends. He completed his assignment on time, and it was a credit to him personally and to Monsanto. At present. Dr. Wright has been given the added assignment of preparing protocols for further toxicity testing of maleic anhydride by CUT. We are confident that he will complete this assignment with equal professional competence. Therefore, it is a pleasure to nominate Dr. Paul L. Wright for a merit award on the basis of his demonstrated performance. G. J. Levinskas I SCM 058800 000--* WATER PCB-SD0000012681 ix m 1. \ I B I T WATER PCB-SD0000012682 Monsanto c . .HIEVE'.IS " 4 Nam* Ol Nomina* Company unit or Staff 0*pt. Oivtflon Mfd. & Env. Hlth. Portion Till* . S+Ury Grade Tnvicoloav Manager Performance 19 0uiin*i Group Annual Salary S31.800 Growth Otpartmtnt Locmon Creve Cceu: Oata of Laft Inc/eate 2/1/76 Date Last Employ* Review Typa of A*d . (Saiact tna awar* Cjt.focy from in* oppottta *ida of ml* tnaat wnlcfl moit appropriattry attend** tn award and antar tna category ano cod* Mlow.) Award Category Technical Accomplishment of significant results Oaicr.ee tn* acnlavemant and It* itgnifleanca to Monaanto Balowi Category Cod* Nun _LD_ At a recent meeting in Creve Coeur, Glenn Schweitzer, head of the Office of Toxic Substances of EPA, offered some interesting comments. He observed that Monsanto's toxicologists were held in high regard at EPA. However, since it lacked an in-house toxicology facility, Monsanto as a company was not as highly regarded overall as duPont, Carbide, or Dow. Dr. Wright's professional and personal characteristics have contributed sig nificantly to Monsanto's image at EPA. He has shown unusual perseverance and dedication, frequently involving his own time, to review and interpret large volumes of data which he subsequently organized for presentation to EPA officials. Particularly noteworthy were his efforts on polychlorinated biphenyls (Aroclors) and chlorinated isocyanurates (ACL products). In the former instance, his excellent analysis and synthesis of widely scattered observations played a prominent role in forestalling EPA's promulgation of unrealistic regulations to limit discharges of polychlorinated biphenyls. EPA's proposed regulations would have precluded the use of these materials by Monsanto's customers. With respect to chlorinated isocyanurates, Dr. Wright has had several contacts with individual scientists at EPA to answer specific questions that they had raised or to inform them of the status of additional studies which, had been undertaken*. Overall, by virtue of the qualities of leadership which Dr. Wright has dis played, he has made an important contribution to Monsanto's image at EPA. That favorable image will become increasingly important to Monsanto as the areas of interaction with EPA multiply. , ^POSITION 1 exhibit , gif r Racommandad Syi (It tv. 10/74) Oat* Award Amount Recommendedi Approved Byi --------- --- Oat* Award Approved for paymentt 7? Aft UfJU.T.Ud SCM 058795 WATER PCB-SD0000012683 E X H I B I T WATER PCB-SD0000012684 'woa i tG * (Rev 4-92) AUTHORIZATION TO RELEASE INFORMATION (PRIVATE PERSON OR ORGANIZATION) TO PROBATION OFFICER TO WHOM IT MAY CONCERN: I.---------------- Paul L. _Wright------------------------------------------ , the undersigned, hereby authorize the United States Probation Office for the EasternDistrict nf Missouri or its authorized representative(s) or employee(s), bearing this release or copy thereof, to obtain anv information in vour files pertaining to my: [if Employment l~~| Education Records (including but not limited to academic achievement, attendance, athletic, personal history, and disciplinary records) Medical Records Psychological and Psychiatric Records ' I hereby direct you to release such information upon request of the bearer. This release is executed with full knowledge and understanding that the information is for the United States Probation Office's official use. I hereby release you, as custodian of such records, any school, college, or university, or other educa tional institution; hospital or other repository of medical records; social service agency, any employer, or retail business establishment including its officers, employees, or related personnel both individually and collectively, from any and all liability for damages ofwhatever kind which may at any time result to me, mv heirs, family, or associates because of compliance with this authorization and request for information or any other attempt to comply with it. The information hereby obtained by the aforementioned probation office is to be used only for the purpose of presentence investigation and report and, if applicable, for supervision. WATER PCB-SD0000012685 P*?O0 140 <3/64) united state* District colt FCOtRAU AKOSa T.On ITITIm REQUEST FOR EMPLOYMENT OATA AOOACSS 0* AROSATION OFFICE 111 U.S. Court & Custom House 1114 Market Street St. Louis, MO 63101-2071 r _ Personnel Department Monsanto Company 800 North Lindbergh Blvd. St. Louis, MO 63141 L 3/7/.Si Dear Sir: The person identified below is j-der investigation by this office. The information requested is needed to complete this investigation. Your cooperation will be greatly appreciated. Please return this form^aiiaia three days in the enclosed anvq * Pat "I namc oe nensprmiNtr^VvcsTiCATCo (L**i WRIGHT. Paul Lee-, aoorem oe rcrsom aaiNC invmtigateo 2001 North Geyer Rd. J St. Louis, MO 63131 ___ 308-34-3416 States employed JovlVWYiV'co'Peb. 29, 1984, as manager of special projects in Dept, of Medical & Environmental Health earning $53,000 per year. Please information oesmeo verify. nCjkSON ron tcamim aTino cmaiotmcmt j f-- ^ ^ >A ,f fi WAS THIS REASON'S SALARY ATTaCnCO' ru (WOULD YOU COWSIOCR AECMRLOYING THIS RCJUONf REMARKS (Cor<mtn4 Ihit pwai'i aifandanea, Y* IWway, wihtmty, and ditttrm* ttamm wplarad tf rw intafuMi.) >T"C AUTHORIZATION TO RELEASE CONFIDENTIAL INFORMATION form attached. 000/.07 WATER PCB-SD0000012686 UJXX / WATER PCB-SD0000012687 GMErt DMEH C" Toxicology Section/St. Louis C0J0tvj0frr7L0CATi0N) SPECIAL (TYPE OF REPORT) REPORT LO N -t CO H ** NO - t2--i Q-CHO CJQ Xa. C=J < </ OoB. Uorn rt cu oi CU tfl oi > CH OX ti--3t >u J4> aO :CiU <H t-j u, o OH 4> ' M>- O U (-> X OUUt-1 Hn CD ft XO vDCi PH 5< O .. IS <P REPORT NO.: MSL-2 002 JOB/PROJECT NO.: DATE: October 14, 1981 TITLE: TOXICITY OF AROCLOR PRODUCTS 1242,;1254 AND 1260 TO THE LIVER OF ALBINO-RATS AUTHORS: George J. Levins leas, Ph.D. abstract: This report is a tabulation of the results of microscopic evaluation of liver sections from rats fed AROCLOR 1242, AROCLOR 1254 or AROCLOR 12 60 for two years. A-IOMIat SCM 058930 0001.03 WATER PCB-SD0000012688 DISTRIBUTION COPY NUMBER 1 - Reports Library, R2C 2 - Reports Library, R2C 3 - Reports Library, R2C 4 - DMEH Library, G2WA 5 - DMEH Library, G2WA 6 - R.T. Berendt, E2ND 7 - J.H. Craddock, A2SA 8 - G.J. Leviriskas, G2WF 9 - J.G. Nassi'f, E2ND '' 10 - J.M. Norris, Dow Chemical 11 - R.A. Stohr, B3NJ ABSTRACT ONLY This report has been assigned to you. When it is no longer needed, you are responsible for returning it to: ____________ REPORTS LIBRARY, R2C_____________ If you transfer it to anyone else, please let your librarian know, so the records can be changed. R-ioaa(C) (Rev. i/ao) ooor.ca ' SCM 058931 WATER PCB-SD0000012689 INTRODUCTION The polychlorinated biphenyls (PCBs) comprise a family of compounds of variable chlorine content. PCBs manufactured by Monsanto (tradenamed "Aroclor") bear a four digit number, the '12' indicating biphenyl and the last two digits indicating the chlorine content by weight percent. Since the chlorine atoms are randomly distributed among the 10 ring positions available for substitution, each material is a mixture of isomers. In 1969, Monsanto sponsored a series of animal studies at Industrial BIO-TEST Laboratories in Northbrook, Illinois, on Aroclor 1242, 1254, and 1260 for the assessment of the health and environmental hazards of these materials. This series consisted of 2-year chronic feeding studies to rats and dogs, a 3-generation rat reproduction study, a rat teratology study, a dominant lethal mutagenic study in mice and a toxicity/reproduction study in chickens on each of the 3 Aroclor products. Even though no such action was contemplated, such a broad battery of tests would have been adequate to support Food Additive Petitions for each of these materials. These studies were initiated by reports that PCBs had been detected in the environment. A report (Nelson, 1972b) by a Panel on Hazardous Trace Substances of an Ad Hoc Committee on Environmental Health Research covers the development of information on the environmental and biological effects of PCBs. There have been subsequent literature reviews which need not be recapitulated here [DHEW, 1978; EPA, 1976; I ARC, 1978; NAS, 1979; Roberts, et al. (1978)]. Starting in 1969, while these studies still were in progress, information regarding them was made available to various government groups.1 Subsequently, data from these studies were presented at a conference -1- OOO'.MO SCM 058932 WATER PCB-SD0000012690 sponsored by the National Institute of Environmental Health Sciences at Rougemont, N.C. on December 20-21, 1971, but the account of these studies was omitted from the published proceedings (Keplinger, et al., 1972; Nelson. 1972a). Subsequently, it was reported that female Sherman strain rats developed hepatocellular carcinomas when fed Aroclor 12602 from Lot No. AK-3; the same lot used for the earlier Monsanto study with this material. As a result, Monsanto requested the contract laboratory's pathologists to review livers from all available rats from the 3 Aroclor studies. That review (which could have included new sections of liver from animals examined previously in addition to sections from animals not examined previously) was presented in the reports of Gordon and Richter (1975a,b,c). In November 1975, reports containing evaluations of liver sections from the 2-year rat feeding studies (Gordon and Richter, 1975a,b,c) and the results of an independent evaluation of the same liver sections (Pour, 1975) were . presented to and discussed with several federal regulatory agencies3. Later that month, a summary of data from all of the studies was presented at the National Conference on Polychlorinated Biphenyls sponsored by the Environmental Protection Agency and held in Chicago on November 19-21, 1975 (Calandra, 1976). Only summaries of data from these and other toxicity studies conducted over the years have been published (Jenkins, et al., 1972; Keplinger, et al.. 1971; Monsanto, 1980). Knowledge of these efforts may have prompted the statement in a recent article that "...Monsanto, whose reaction to the possibility that polychlorinated biphenyls (PCBs) might be an ecological disaster was a classic of how such issues should be handled, says Ford4. Monsanto began to investigate the dangers as soon as they were 2- SCH WATER PCB-SD0000012691 seriously voiced. It insisted on keeping an open mind about them. As soon as evidence appeared that there was a strong chance PCBs were a hazard, it published the results of its investigations, admitting the danger. It thus established a reputation of honesty even among the environmentalists." (Clutterbuck 1981). At a later meeting in Bethesda, MD.5, pathologists selected and reviewed some liver slides from the Monsanto-sponsored and Kimbrough studies. The pathologists differed in the terminology used to classify the lesions. They did conclude that the general incidence and severity of liver lesions (hyperplasia, nodular hyperplasia and neoplastic changes - carcinomas) were greater in the rats from the study subsequently published by Kimbrough, et al. (1975). No carcinomas were seen in the Monsanto-sponsored studies. It was noted that either the strain of rat or sex could have accounted for the differences. The spontaneous incidence of hyperplastic or neoplastic liver lesions in the Sherman strain rat was unknown, and the occurrence of such lesions appears to be higher in female rats and mice. The different diagnoses were discussed with Dr. Philippe Shubik of the Eppley Institute for Research in Cancer at the University of Nebraska. Dr. Shubik suggested that the liver sections from these studies could be reviewed by Dr. Parvis Pour. This was done. This publication is an effort to make readily available information in the Gordon and Richter reports (1975a,b,c) which are only summarized in the literature (Calandra, 1976). - 3 SCH OSS^ 000 O A *+* WATER PCB-SD0000012692 METHODS It appears that the Threshold Limit Value of 0.5 mg'/m3 for chlorobiphenyl with 54% chlorine (ACGIH, 1969) was used to estimate suitable dose levels for the rat feeding studies. For that purpose the following assumptions were made: if the ambient air concentration of PCBs is 0.5 mg/m3, and if all of the inhaled PCBs are absorbed, and if 15 m3 of air are inhaled during the working day, a person would receive a PCB dose of 7.5 mg/day. The corresponding dosage would be approximately 0.1 mg/kg/day for a 70-kilogram person. Consequently, dosages of 0.1, 1, and 10 mgAg/day were decided upon, and the dietary concentrations were set at 1, 10, and 100 ppm. As it turned out, the assumptions used to set dosages for the feeding study resulted in the low dose level rats receiving a dosage that was 100 times higher than the 0.001 mg/kg/day which FDA later calculated as allowable for protracted ingestion based on human data (FDA, 1973). For the chronic toxicity study in rats6, weanling animals of the Charles River CD strain7 were divided into a control group and 9 treatment groups, each consisting of 50 males and 50 females, with 3 treatment groups assigned to each of the 3 Aroclor products studied (1242, 1254, and 1260). Not all of these animals started at the same time. After about 2 months on test, additional groups of males and females were assigned to each test diet and the controls. These animals were added to allow a sufficient number of animals for sacrifices at 3, 6 and 12 months to provide some information prior to the completion of the 2-year study period. The numbering-sequence SCM 058935 OOO: '.3 WATER PCB-SD0O00012693 and^-the.^4esicj.ation,- of . some. ,.an tmals^as-.JlExtra-"-~suggests that additional animals were, placj'd^on test. Groups of 5 males and 5 females were scheduled to be sacrificed after 3, 6, and 12 months of feeding1, and survivors were to be sacrificed at the end of the test period. Animals were to be given a gross autopsy and tissues were to be preserved for possible histologic examination. suggestive of tumors were to be examined. Tumors or lesions RESULTS Data from the Gordon and Richter reports (1975a,b,c) on liver slides are shown in Tables 1, 2, and 3 for Aroclor 1242, 1254, and 1260, respectively. While the text of the reports contained comments specific to the particular Aroclor, they also contained similar comments such as "There is evidence of a chemical effect on the liver...........This consists of a hepatocellular alteration beginning as focal hypertrophy which progresses to nodular hyperplasia, and in a few animals to hepatoma or cholangiohepatoma" and "In the absence of metastasis, invasiveness, severe basophilia, mitoses, or other evidence of anaplasia; these are benign tumors rather than malignant carcinomas. There was no evidence of metastasis or invasiveness of these tumors in this study". The reports also stated that "The other treatment-related lesions reported are regarded as degenerative or hyperplastic in nature and they are morphologic manifestations of an adaptive response of the liver associated with biotrans formation of the test material" and "In most instances, the spectrum of treatment-related histopathological findings in the liver from this re-evaluation did not differ significantly from that previously reported in our original report...........No hepatocellular carcinomas were observed". '5" ` SCM 058936 <no:u* WATER PCB-SD0000012694 With respect to Aroclor 1254, it was noted that "One liver tumor (a hepatoma) previously reported in a T-II animal (No. 445) was re-classified as nodular hyperplasia" (Gordon and Richter, 1975a)10. DISCUSSION The initial reports of these studies concluded that the target organ was the liver for each Aroclor product. This was confirmed by the second evaluation of liver slides (Gordon and Richter, 1975a,b,c). These alterations were slow to develop, their incidence being related to both dose level and duration of treatment. Since there were increased incidences of vacuolar changes in the cytoplasm of the hepatocytes and focal hypertrophy at all dose levels for all 3 Aroclor products at the 24-month sacrifice, a "no-effect" level was not established for liver effects. With respect to the slides from Industrial BIO-TEST, Pour (1975) concluded that Aroclor 1242, 1254, and 1260 "showed a dose-dependent hepatotoxic effect, characterized by degenerative and regenerative processes. With one exception, all lesions were considered non-neoplastic. Structures similar to cholangiocarcinomas and hepatomas were found in one rat. However, the possibility of metastases of a carcinoma into the liver has been also considered." In the report, he noted one lesion which seemed to "represent a metastatic tumor (probably a mammary gland carcinoma of the same animal from which ...[the particular liver section]... was submitted), rather than a cholangiocarcinoma". This occurred in an animal fed 100 ppm of Aroclor 1254. The contract laboratory pathologists diagnosed the presence of mammary tumor metastases in the liver of this rat (Gordon and Richter, 1975b).11 SCM 058937 (no:'.:5 WATER PCB-SD0000012695 SUMMARY and CONCLUSIONS Groups of male and female rats were fed diets containing either 1, 10, or 100 ppm of one of 3 Aroclor products, 1242, 1254, or 1260, for 2 years. Focal hypertrophy of the liver occurred at 3, 3, and 12 months for rats fed Aroclor 1260, 1254, and 1242, respectively. Focal hypertrophy was not seen in livers of control animals. At the 24-month sacrifice, livers of animals from all dose levels of the 3 Aroclor products had an increased incidence of vacuolar changes and focal hypertrophy. Livers of some animals fed 100 ppm of each Aroclor also had benign liver tumors (hepatoma or cholangiohepatoma). No hepatocellular carcinomas were observed. 7- SCH 058938 ono WATER PCB-SD0000012696 Footnotes Dates of initial correspondence and contacts. October 3, 1969. E.P. Wheeler to A.R. Glasgow, Division of Pesticides. Food and Drug Administration. April 8, 1970. R.E. Kelly to H. Bluraenthal, Petitions Review Branch, Food and Drug Administration. ' April 22, 1970. E.P. Wheeler to E.J. Burger, Jr., Office of Science and Technology. June 1, 1970. E.P. Wheeler to H.E. Stokinger, Bureau of Occupational Safety and Health. R.D. Kimbrough, personal communication. November 13-14, 1975. Council on Environmental Quality. Environmental Protection Agency. Food and Drug Administration. House Subcommittee Manpower, Compensation, Health and Safety. National Cancer Institute. National Institute for Environmental Health Sciences. National Institute for Occupational Safety and Health. Dr. David Ford of Bath University. . At National Cancer Institute on January 21, 1975. Present were D.E. Gordon, R.D. Kimbrough, G.J. Levinskas, R.A. Squire, W.R. Richter. .' Since the events described earlier, the validity of many toxicity studies conducted by Industrial BIO-TEST Laboratories has been challenged. Therefore, the available raw data supplied by Industrial BIO-TEST was reviewed to determine whether this study could be validated. The review showed that the data bases (including the lack of a protocol) were insufficient for a complete validation of the study. It was decided to focus on presenting the primary liver effects reported by Gordon and Richter (1975a,b,c). Therefore, a complete audit was not undertaken. Available records were examined for a determination that the animals were placed on test and of their ultimate fate. Necropsy and microscopic reports were also examined for findings pertaining to livers of those animals. Significant discrepancies which were found between data in Tables 1, 2, and 3 and the data base for the Gordon and Richter reports are noted in this report. On some records, the labels Aroclor 1242 and Aroclor 1260 are interchanged as determined by a check of the animal numbers. Charles River Breeding Laboratories, North Wilmington, Massachusetts. Animals sacrificed at 6 and 12 months were placed on test about 2 months after the first group. Those sacrifice periods are sometimes labeled 8 and 14 months, respectively, which corresponds to the calendar time from the start of the test but overstates the time on test for those groups. 8SC* ooo 17 WATER PCB-SD0000012697 9. As a result of the examination described in footnote 6, a few animals were reassigned to other dose levels on basis of assigned numbers. Three with no recorded liver lesions were deleted from the 24-month listing because of short duration on test: 14 months at 100 ppm Aroclor 1242, 15 months at 10 ppm Aroclor 1254 and 13 months at 100 ppm Aroclor 1254. Therefore, numbers in Tables vary slightly from those in reports of Gordon and Richter (1975a,b,c). 10. That change and comments in the footnotes to the Tables were noted ' during the examination described in footnote 6. In addition, the following also were noted during the examination. Aroclor 1254 - 100 ppm animal marked "Extra3" with diagnosis of hepatoma in record of examination for original Industrial BIO-TEST report. Animal not listed in Gordon and Richter report. . - 100 ppm animal no. 542 with gross autopsy notation that liver appears tumorous and white foci has no record of examination. Aroclor 1260 - 100 ppm animal no. 293 with gross autopsy notation of tumor on liver has no record of examination. . In some instances, diagnoses were crossed out on the available records. These were included in the Tables. Crossed out diagnoses for hepatoma or cholangiohepatoma are noted in the footnotes to the Tables. ' 11. Dr. Pour's report does not include the following liver sections noted by discrepancies between his tabulation and the Gordon and Richter reports. 1 from control at 12 month sacrifice, 5 from 100 ppm Aroclor 1242 at 24 months, 1 from 100 ppm Aroclor 1260 at 3 months. None of these animals were reported as having hepatomas or cholangiohepatomas in the Gordon and Richter reports. SCM 058940 nnn''^ WATER PCB-SD0000012698 References 1. ACGIH (1969). Threshold Limit Values of Airborne Contaminants. American Conference of Governmental Industrial Hygienists. Cincinnati. 2. Calandra, J.C. (1976). Summary of toxicological studies on commercial PCB's. In: National Conference on Polychlorinated Biphenyls, November 19-21, 1975. Chicago, Illinois. EPA Report No. 560/6-75-004. March. . NTIS PB-253 248. pp.35-42. 3. Clutterback, D. (1981). What causes environmental conflict? New Scientist 90, 176-177. 4. DHEW (1978). Final Report of the Subcommittee on Health Effects of PCBs and PBBs. Environ. Health Perspect., 24, 129-198. 5. EPA (1976). National Conference on Polychlorinated Biphenyls, November 19-21, 1975. Chicago, Illinois. U.S. Environmental Protection Agency. Washington, D.C. EPA-560/6-75-004, March. NTIS PB-253 248. 6. FDA (1973). Polychlorinated biphenyls (PCB's). Contamination of animal feeds, foods and food-packaging materials. Fed. Regis., July 6, 38, 18096-18103. 7. Gordon, D.E. and Richter, W.R. (1975a). Two-year chronic and toxicity study with Aroclor 1242 in albino rats. Histopathological evaluation of additional liver sections. March 24 (unpublished observations). 8. Gordon, D.E. and Richter, W.R. (1975b). Two-year chronic oral toxicity study with Aroclor 1254 in albino rats. Histopathological evaluation of additional liver sections. March 24 (unpublished observations). 9. Gordon, D.E. and Richter, W.R. (1975c). Two-year chronic oral toxicity study with Aroclor 1260 in albino rats. Histopathological evaluation of additional liver sections. March 24 (unpublished observations). 10. IARC (1978). Polychlorinated biphenyls. IARC Monographs on the evaluation of the carcinogenic risk of chemicals to humans. 18, 43-103. International Agency for Research on Cancer. Lyon, France. October. 11. Jenkins, D.H., Keplinger, M.L., Fancher, O.E., Wheeler, E.P. and Calandra, J.C. (1972). Reproductive effects of polychlorinated biphenyls in white leghorn chickens. Toxicol. Appl. Pharmacol. 22, 316. 12. Keplinger, M.L., Fancher, O.E., Calandra, J.C. (1971). Toxicologic studies with polychlorinated biphenyls. Toxicol. Appl. Pharmacol. 19, 402-403. - 10 - SCM 058941 /t> * r\ WATER PCB-SD0000012699 13. Keplinger, M.L., Fancher, O.E., Calandra, J.C., et al. (1972). Toxicological studies with polychlorinated biphenyls. Read at PCB Conference, Quail Roost Conference Center, Rougemont, North Carolina, 20-21 December 1971. Cited in Polychlorinated Biphenyls Environmental Impact. A Review by the Panel on Hazardous Trace Substances. March 1972. Environ. Res. 5, 249-362. 14. Kimbrough, R.D., Squire, R.A., Linder, R.E., Strandberg, J.D., - Montali, R.J., and Burse, V.W. (1975). Induction of liver tumors in Sherman Strain female rats by polychlorinated biphenyl Aroclor 1260. J. Natl. Cancer Inst. 55, 1453-1459. 15. Monsanto Company (1980). Polychlorinated biphenyls. PCB's. A report on Uses, Environmental and Health Effects and Disposal. 16. NAS (1979). Polychlorinated biphenyls. National Academy of Sciences. Washington, D.C. 17. Nelson, N. (1972a). Comments on research needs. Environ. Health Perspect., 1, 181-185. 18. Nelson, N. (1972b). Chairman, Panel on Hazardous Trace Substances. Polychlorinated biphenyls - environmental impact. Environ. Res. 5, 249-362. 19. Pour, P. (1975). Histopathological reevaluation of livers for rats treated with Aroclor. August 1 (unpublished observations). 20. Roberts, J.R., Rodgers, D.W., Bailey, J.A., Rorke, M.A. (1978). Polychlorinated Biphenyls: Biological criteria for an assessment of their effects on environmental quality. National Research Council of Canada. Ottawa. Publication No. NRCC 16077. - 11 - cnQ..~o SCH WATER PCB-SD0000012700 Sacrifice Interval Diet Level (ppm) Table 1 Aroclor 12A2: 'Primary Liver Lesions Among Albino Rats 3 Months 0 1 10 100 6 Months 0 1 10 100 12 Months 0 1 10 100 Termina1 a 0 1 10 100 No. Animals Examined Findings Vacuolar change Focal necrosis Focal lymphoid infiltration Focal hypertrophy hepatocytes Nodular hyperplasia Ductular hyperplasia . Hepatoma Chola ngiohepa toma 10 8 10 9 202 010 000 000 00 0 00 0 000 000 1 0 0 0 0 1 0 0 10 10 8 9 1 00 1 10 030 000 000 000 000 000 0 0 0 0 0 0 0 0 10 10 10 9 1 2A 000 00 1 000 000 101 000 000 0 0 0 1 0 0 0 0 23 32 29 19 178 111 1 00 023 111 533 000 000 9 0 0 8 8 3 3b lc * Control group has animal dead at 19 months and 2 marked "Extra". 1 ppm group has animals dead at 19, 22, 22, 23, 23 months. 10 ppm group has aniaMls dead at 23, 23 months. ,t100 ppm group has animals dead at 22, 22, 22 months and 2 marked "Extra". b Includes 1 animal without record of examination in original IBT report. Not listed as hepatoma in worksheets for Gordon and Richter report but appears and was crossed out on typed tabulation for animal marked "Extra". q for other 2 animals do not appear in records of examinations for original IBT report. O' ' Q Diagnosis does not appear in records of examinations for origina'l IBT report. t; Diagnoses SCH 0 5 8 9 ^3 WATER PCB-SD0000012701 Sacrifice Interval Diet Level (ppm) Table 2 Aroclor 1254: Primary Liver Lesions Among Albino Rats 3 Months 0 1 10 100 6 Months 0 1 10 100 12 Months 0 1 10 100 Terminal a 0 1 10 100 No. Animals Examined Findings Vacuolar change Focal necrosis Focal lymphoid infiltration Focal hypertrophy hepatocytes Nodular hyperplasia Ductular hyperplasia Hepatoma Cholangiobepatoma 10 10 10 10 233 01 1 000 000 000 000 000 000 1 0 1 1 0 0 0 0 10 10 10 10 1 00 1 20 000 000 000 000 000 000 0 2 2 4 0 0 0 0 10 10 10 10 1 14 0 00 002 004 000 111 000 000 1 1 1 2 0 0 0 0 23 30 26 26 1 7 9 13 131 1 120 1 0 3 4 14 1 0 3 14 563 00 0 4 4b 000 2C a Control group has animal dead at 19 months and 2 marked "Extra". 1 ppm group has animals dead at 22, 22 amnths, 1 marked "Extra" and 1 other for which date of death is not recorded. 10 ppm group has animals dead at 22, 22, 22 months. ,,100 ppm group has animals dead at 23, 23 months and 9 marked "Extra". b Includes 2 animals marked "Extra". Diagnoses do not appear in records of examination for original IBT reports. Qc Both animals marked "Extra" with same designation. 1 without apparent record of examination for original IBT report. q Diagnosis for other animal does not appear in records of examinations for original IBT report. r: 11 os, WATER PCB-SD0000012702 Table 3 Aroclor 1260: Primary Liver Lesions Among Albino Rats Sacrifice Interval Diet Level (ppm) 3 Months 0 1 10 100 6 Months 0 1 10 100 12 Months 0 1 10 100 Terminal^ 0 1 10 100 No. Animals Examined Findings 10 10 iob 10 10 6 5 9 10 9 10 9 23 26 25 25 Vacuolar change 202 3 11 Focal necrosis 0 0 4C 0 10 Focal lymphoid infiltration 000 0 02 Focal hypertrophy hepatocytes 000 2 00 Nodular hyperplasia 000 0 00 Ductular hyperplasia 00 1 0 00 Hepatoma 000 0 00 Cholangiohepatoma 000 0 00 *. Control group has animal dead at 19 months and 2 marked "Extra". 2 ld 2 0 0 0 0 0 6 0 0 3 0 0 0 0 1 25 000 000 000 0 00 1 10 000 000 3 0 0 4 1 2 0 0 157 9 143 6 10 1 0 0 3 13 9 107 6 5 6 7 12 0 0 le 7f.g 00 0 1 ppm group has animals dead at 20, 22, 22 months. 10 ppm group has animals dead at 19, 22, 22 months and 1 marked "Extra". 100 ppm group has animals dead at 22, 22 months. ** Includes 1 marked "Extra". c Worksheets show listings under this diagnosis with arrow pointing to Vacuolar change column. d,Date of death of this animal not recorded. e No record of examination for original IBT report. Listed on worksheets for Gordon and Richter report with diagnosis crossed out. f Includes 2 animals without record of examination for original IBT report. Diagnoses for other 5 animals do not appear in records of examinations for original IBT report. 2 of these diagnoses crossed out on worksheets for Gordon and Richter reports. Includes one animal with both diagnoses. Hepatoma is 1 of 2 crosaed out (superscript f). ^ h Includes 3 animals without record of examination for original IBT report. Diagnosis for other animal does not appear in . records of examinations for original IBT report. 2 of these diagnoses crossed out on worksheets for Gordon and Richter ^ reports. u' WATER PCB-SD0000012703 S 7 0 B i> U W J i U JX X I B I T ft 41 7 WATER PCB-SD0000012704 jruhuiual B 1 O * T E 5 T Jne. -` * ' . REPORT TO MONSANTO COMPANY . TWO - YEAR CHRONIC ORAL TOXICITY STUDY WITH ARCCLOR 1254 - IN ALBINO RATS HISTOPATHOLOGICAL EVALUATION OF ADDITIONAL LIVER SECTIONS MARCH 24, 1975 * IBT NO. 641 - 06672 ' . - L Introduction . At the request of Dr. Levinskas of the Monsanto Company, additional sections of liver from a two - year chronic oral toxicity^ study of Aroclor 1254 in rats (IBT No. 622-07298) were processed into H Sc E stained sections and evaluated by light microscopy. Ths following report presents the results of this study. . GOQ:'~l WATER PCB-SD0000012705 Jnduiixlal B I O T E S T }rtc. 2 II. Summary In most instances, the spectrum of treatment-related histopathological findings in the liver from this re-evaluation did not differ significantly from that previously reported in our original report dated November 12, 1971. There were six hepatomas detected among six of the animals at the highest treatment level (100 ppm) of the 24-Month Sacrifice. No hepatocellular carcinomas were observed. One liver tumor (a hepatoma) previously reported . in a T-II animal (No. 445) was re-classified as nodular hyperplasia. The . ' other treatment-related lesions reported are regarded as degenerative or . ' hyperplastic in nature and they are morphologic manifestations of an adaptive ' . - response of the liver ascribed to biotransformation of the test material. The . Hatter-lesions wie confined primarily to lest animals of the 12 and 24 nor.-, sacrifices and they were dose-related in incidence and severity. * f?r:: In conclusion, Aroclor 1254 does not appear to be carcinogenic in rats ; fed. for ;two years at levels up to and including 100 ppm. . Respectfully submitted, INDUSTRIAL BIO-TEST LABORATORIES, INC. * Report Prepared and Reviewed by: D. E. Gordon, D.V. M. ,Ph. D Section Head, Pathology Report Approved by: M. L. Kepli/ger, Jpb, D; Manager, Toxicology WATER PCB-SD0000012706 JntLulxlai B l O - T 6 5 T IBT No. 622-07298 Monsanto 3no. 3 I have examined additional sections of liver from rats of IBT No. 622-07298 and have re-evaluated them for evidence of carcinogenesis. I have tabulated my findings for Arodor 1254 on the following pages: There is evidence of a chemical effect on the liver which is most pronounced at time of the 12-Month and 24-Month sacrifice. This consists of a hepatocellular alteration beginning as focal hypertrophy which progresses to nodular hyperplasia, and in a few animals to hepatoma or cholangiohepatoma. The hypertrophic cells *?.P.; 4$ contain large amounts of light staining cytoplasm which is probably rich in glycogen and endoplasmic reticulum. In some cases ring shaped structures, probably representing whorls of proliferative endoplasmic reticulum, were seen v in the cytoplasm of these cells. . The hyperplastic nodules occupied larger areas and often compressed' surrounding cells of the normal liver parenchyma. They also contained the same hypertrophic cells. ' ' '' ` * Those lesions classified as hepatomas or cholangiohepatomas were larger nodules which showed evidence of confluence or some variation in cell size, shape, staining or a ductular or adenomatous pattern of growth. In the absence .of metastasis, invasiveness, severe basophilia, mitoses, or other evidence of anaplasia; these are benign tumors rather than malignant carcinomas. There was no evidence of metastasis or invasiveness of these tumors in this study. SCH 058168 000 *vb WATER PCB-SD0000012707 JuduiiniaL B I O * T E S T abyiala'u*&, 3*c. 4 /With. exception of the vacuolar changes in the cytoplasm of hepatocytes, the more severe hepatocellular alterations were confined to animals of the 24Month Sacrifice. Ward R. Richter, D. V. M., M. S. Dipl., Am. College Vet. Path. SCM 058169 O'lQ: .^7 WATER PCB-SD0000012708 5 ' Lesion Primary Liver Lesions - Aroclor 1254 3 Month Sacrifice - Rata CroHio TI th Tin Control Vacuolar Change Focal Hypertrophy Nodular Hyperplasia Hepatoma Ductular Hyperplasia Cholangio -Hepatoma Hepatocellular Necrosis 3/10 0/10 0/10 0/10 0/10 0/10 1/10 3/10 0/10 0/10 0/10 0/10 0/10 1/10 1/10 1/10 0/10 0/10 0/10 0/10 0/10 2/10 ...... 0/10 0/10 0/10 0/10 0/10 0/10 SC* ooo::.-3 WATER PCB-SD0000012709 6 Primary Liver Lesions - Aroclor 1254 6 Month Sacrifice Rats Lesion Vacuolar Change Focal Hypertrophy Nodular Hyperplasia Hepatoma Duetular Hyperplasia Cholangio-Hepatoma Hepatocellular Necrosis TI 0/10 0/10 0/10 0/10 0/10 . 0/10 2/10 th 0/10 0/10 0/10 0/10 0/10 0/10 .0/10 Grouo Tin 1/10 4/10 0/10 0/10 0/10 0/10 1/10 Control 1/10 0/10 0/10 0/10 0/100/10 1/10 q58 sew WATER PCB-SD0000012710 7 Primary Liver Lesions - Aroclor 1254 12 Month Sacrifice - Rats Lesion Vacuolar Change Focal Hypertrophy Nodular Hyperplasia Hepatoma Ductular Hyperplasia Cho langio -Hepatoma Hepatocellular Necrosis H 1/10 0/10 o/io 0/10 1/10 0/10 0/10 th 4/10 4/10 0/10 0/10 - . 1/10 0/10 0/10 Grouo Tin Control 1/10 1/10 2/10 0/10 0/10 0/10 0/10 0/10 0/10 1/10 0/10 0/10 1/10 0/10 sc 5Sin ooo:' "0 WATER PCB-SD0000012711 8 Primary Liver Lesions - Arocior 1254 Terminal Sacrifice - Hats % Lesion Vacuolar Change Focal Hypertrophy Nodular Hyperplasia Hepatoma Ductular Hyperplasia Cholangio-Hepatoma Hepatocellular Necrosis TI 8/31 3/31 0/31 0/31 6/31 0/31 3/31 TII 10/26 5/26 3/26 0/26 3/26 . 0/26 1/26 Croup TUI 13/27 13/27 13/27 4/27 4/27 2/27 2/27 Control 1/23 0/23 1/23 0/23 5/23 0/23 1/23 SCH 058U3 ooq:: 31 WATER PCB-SD0000012712 1. 1 1 1 1 t1 TflUOW j -- -n Dose Level * \ V^V-J.*WU T a b u la tio n of In d iv id u a l L iv e r Lesions in Rats _ ------ ------,, in >JA zfHt *2-- nA *-- * * - oo U .o dW ` il1 * *o t,tf * Bd 0m : *gd tJ 1* 0 #- ' - - UOOSI'(USU0IOIUO=II'IUlU=UIUIlhUOIU|ItNSOIUhOSIhUOS'IU>h5IjUOmgI' UOS <*=JO*DSJI*2i4JhO*JJ'Oi|=w<OU=(lU0=wI2OU^> 1 a0Otwk0O(uv0O >wkOj0u^w0cr*<wUa1utitnlw-l4ilw0U1' ZZZZh)ZZZZg tn x K aa. 2 ro 2 a3 + *<:+ + + -j- Cytoplasmic vacuolation of hcpatocytes ++ + Focal necrosis of hcpatocytes . Focal lymphoid infiltrations .. Focal hypertrophy of hcpatocytes . '. t ., . Nodular hyperplasia of hcpatocytes Focal bile duct hyperplasia Hupatoma % . r <a t r u 30u Cholanglo-hepatoma 1. 1 i SCM 1 OGO o VA C*--D r f . .* ....... Squamous Cell Carcinoma, metastatic Mammary tumor, metastatic lie pat it is /Ci r rlios i a 1 WATER PCB-SD0000012713 AHUULUK - 1<134 Tabulation of Individual Liver Lesions in Rats acrifica 1 erval Dose Level - Anima No. and Sex * M OO >2 U AQ. c JaS o o o2 .92) O c2uU s3Ac'. nu a a oc a* O~ 12 Ss* =w --_ C 1u3 o > y___ b fc. Liver Lesions o 2* c.2 ou >y -Oaa wioas. x>. xo yIS o cZ IS _ 22 g-* oa. o --5* ias bo is -- l o. Z U is oo 3| IuS ioZ 4a 9 X * H 990Q. X1 Q M C o U 6| vfonth. I B-I lppm 921M 922" 923" 924" 925" 936F 937" 938" 939" 940" B-H lOppm 950M 951" 952" 953" 954" 966F 967" 968" 969" 970" B-in 981M lOOppm 982" 983" 984" 985" 996F # 997" 998" 999" 1000" + + + + + + r SCM 058175 Squamous C e ll C arcinom a, m e ta sta tic iu ir*n WATER PCB-SD0000012714 S a c rific e I aterval i ( i I A R O C LO R - l^S -t T a b u la tio n of In d ivid u a l L iv e r Lesions in Rats L iv e r Lesions ` . ... . .. -. **'*>#' ' * if Dose Level - SB 0 OiOM'IIMn^UMi. :ss:N|:s!<^ X + +. . S9 o 1~ y OkUIUIUIUIUIUIUIUIW 040)>40'UlitwUNM . , O^U'JIMlI^UNi-1 + + XX . + ++ . > Oi P 2i E. x a. ? 3 * Cytoplasmic vacuolation of hepatocytcs Focal necrosis of hepatocytes Focal lymphoid infiltrations ++++++ . Focal hypertrophy of 1 hepatocytes Nodular hyperplasia * of hopatocytes Focal bile duct . `+ . - hyperplasia 4 Hepatoma i \ Cholanglo-hepatoma 1 i. ! SCM 1 o \J\ 00 -4 ---------- O' 4. . . . Squamous Ceil 1 Carcinoma, metastatic Mammary tumor, ' metastatic 1 WATER PCB-SDOOOO012715 onorc WATER.PCB-SD0000012716 i d o u u tio n ot m a iv ia u ii u v e r Lesions in R ats Liver Lesions * ........ * > _______ . _ __N __ ___ ___; __b* w -1 g s fr Th*Sp--S hnHA* Dose Level 1 i s? i o^o^u^iua1>'Ii-^>OO'v0o'UUIvUoiOUi'INIItU->IUoh^oOinOuONM<O'iOM'NUN.ohixh)-hvihui > wx nea. r^o2Dpm. Cytoplasmic vacuolation of hepatocytes + Focal necrosis of hepatocytes Focal lymphoid Infiltrations + t+ t Focal hypertrophy of hepatocytes 10 . *d u Nodular hyperplasia . of hepatocytes .' . + .t Focal bile duct '+ ityporplasla Hepatoma b Cholanglo-hepatoma i i i i i i SCM oU> *0-0 a> . , . .. , >0 Squamous Cell Carcinoma, metastatic Mammary tumor, i metastatic WATER PCB-SDOOOO012717 A U U C M I K - I <154 T a b u la tio n of In d iv id u a l L iv e r Lesions in Rats = m ild in severity = m oderate ia se ve rity .m 4* + 01 p (HaX*. trt < u ae tv> o & Oo W, 30 B U1U1UIU1UIi|ki|kUIU1UIU1i^i^^i^ ll Sl L il1 u W U ObLtLilk- 4`^ N.........w........N... o--o o"o H M O O vO vO vO 00 552 = Z Z S -Ot U to h* = = z rhj r Z z z vO = *4 S >D S *4 = = 4- 4 itt-t -- - -..-KH L ^ H* *p- n n ro< OWo aa cx"n "aap. ^ 2 B p Cytoplasmic vacuolation ' of hepatocytes Focal necrosis of hepatocytes Focal lymphoid infiltrations 1 1 '+ 11 + + + t +t tX + d *d *d *d . *d *d *d' *d*d + t+ *d *d *d *d Focal hypertrophy of hepatocytes Nodular hyperplasia i of hepatocytes Focal bile duct -t-' hyperplasia <st n Uol o OB O \ o 1 *d d d d d d J-Iepatoina d Cholangio-hepatoma Squamous Cell Carcinoma, metastatic Mammary tumor, metastatic WATER PCB-SD0000012718 LUX X \ \- I B I T ii i WATER PCB-SDOOOO012719 .t !)n&u&foud BIO-TEST Jk/xyi&tyiie&. Snc. 1810 FRONTAGE ROAO ,, NORTHBROOK, ILLINOIS 60082 REFORT.TO MONSANTO COMPANY TWO - YEAR CHRONIC ORAL TOXICITY STUDY WITH AROCLOR 1260 IN ALBINO RATS HISTOPATHOLOGICAL EVALUATION OF ADDITIONAL LIVER SECTIONS . MARCH 24, 1975 IBT NO. 641 - 06672 000 33 WATER PCB-SD0000012720 Jndx-iiuaJ. BIO-TEST ^aIn>\a-L>\ci, 3*g. REPORT TO ( ` MONSANTO COMPANY TWO - YEAR CHRONIC ORAL TOXICITY STUDY WITH AROCLOR 1260 -: _ IN ALBINO RATS HISTOPATHOLOGICAL EVALUATION OF ADDITIONAL LIVER SECTIONS : march 24, _ 1975 IBT NO. 641 - 06672 . L Introduction . . - . At the request of Dr. Levinskas of the Monsanto Company, additional sections of liver from a two - year chronic oral toxicity study of Aroclor 1260 in rats (IBT No. 622-07298) were processed into H te E stained sections and evaluated by light microscopy. The following report presents the results of this study. SCN O50181 000 o .0 0 WATER PCB-SD0000012721 fndnilMal B I O - T E S T JaJxyialo'Jmc. \ 2 II. Summary In most instances, the spectrum of treatment-related histopathological findings in the liver from this re-evaluation did not differ significantly from that previously reported in our original report dated November 12, 1971. ' .There were seven hepatomas detected among seven of the animals at the highest treatment level (100 ppm) of the 24-Month Sacrifice. No hepatocellular carcinomas were observed. The other treatment-related lesions reported are regarded as degenerative or hyperplastic to nature and they are morphologic manifestations of an adaptive response of the liver associated with biotransformation of the test material. In general, the latter findings were confined primarily to test animals of the 6-, 12- and 24-Month Sacrifices, and they were dose- related in incidence and severity. . In conclusion, Aroclor 1260 does not appear to be carcinogenic in rats fed for two years at levels up to and including 100 ppm. . Respectfully submitted, . INDUSTRIAL BIO-TEST LABORATORIES, INC. Report Prepared and Reviewed by: D. E. Gordon, D.V.M. ,Ph. D. Section Head, Pathology Report Approved by: M. L. Kepiin^er, Pb^O. Manager, Toxicology SCM 058182 Q^O ;o WATER PCB-SD0000012722 jnduii/Ual B I O * T E 5 T $*<a. 3 IBT No. 622-07298 Monsanto I .have examined additional sections of liver from rats of IBT No. 622-07298 and have re-evaluated them for evidence of carcinogenesis. I have tabulated - my findings for Aroclor 1260 on the following pages: There is evidence of a chemical effect on the liver which was most evident at time of the 12-Month and terminal (24-Month) sacrifices. This consists of a hepatocellular alteration beginning as focal hypertrophy which progresses to nodular hyperplasia, and in a few animals, to hepatoma or cholangiohepatoma. The hypertrophic cells contain large amounts of light staining cytoplasm which is probably rich in glycogen and endoplasmic reticulum. In some cases, ring shaped structures, probably representing whorls of proliferative endoplasmic reticulum, were seen in the cytoplasm of these cells. The hyperplastic nodules occupied larger areas and often compressed surrounding cells of the normal liver parenchyma. They also contained the same hypertrophic cells. Those lesions classified as hepatomas or cholangiohepatomas were larger nodules which showed evidence of confluence or some variation in cell size, shape, staining or a ductular or adenomatous pattern of growth. In the absence of metastasis, invasiveness, severe basophilia, mitoses or other evidence of anaplasia, these are benign tumors rather than malignant carcinomas. There was no evidence of metastasis or invasiveness of these , tumors in this study. SCM 058183 000 41 WATER PCB-SD0000012723 jridtiiUlaL B I O T E 5 T Ja&Moiyuu. Jnc. 4 With exception of the vacuolar changes in the cytoplasm of hepatocytea, the more severe hepatocellular alterations were confined to animals of the 12 % and 24-Month Sacrifice periods. ' (Jcuiej'R __________ Ward R. Richter. D. V. M. , M. 3. Diplocnate. American College of Veterinary Pathologists so 5818`' 000 1 * V^ WATER PCB-SD0000012724 5 . Primary Liver Lesions - Aroclor 1260 3 Month Sacrifice - Rats Lesion . - Group TI TII 'tiii ' Vacuolar Change 0/11 4/10 3/10 Focal Hypertrophy 0/11 0/10 2/10 Nodular Hyperplasia 0/11 0/10 0/10 Hepatoma . 0/11 0/.10 0/10 Ductular Hyperplasia 0/11 . 1/10 0/10 Cholangio -Hepatoma 0/11 0/10 0/10 Hepatocellular Necrosis 0/11 0/10 0/10 Control 2/10 0/10 0/10 0/10 0/10 a/10 0/10 GOO:*.43 WATER PCB-SD0000012725 6 Primary Liver Lesions - Aroclor 1260 ------- 6 Month Sacrifice - Rats -- Lesion Group - TI TII T1H Vacuolar Change 1/6 2/5 5/9 Focal Hypertrophy 0/6 0/5 3/9 Nodular Hyperplasia 0/6 0/5 0/9 Hepatoma ' 0/6 0/5 0/9 Duetula r Hyperplasia 0/6 0/5 0/9 Cholangio-Hepatoma 0/6 0/5 0/9 Hepatocellular Necrosis 0/6 1/5 0/9 Control 1/10 0/100/10 0/10 . 0/10 0/10 1/10 SCM 058186 ooo:-.M WATER PCB-SD0000012726 7 Primary Liver Lesions - Aroclor 1260 12 Month Sacrifice - Rats Lesion ' GrouD TI TQ Till Vacuolar Change 2/9 5/10 3/9 Focal Hypertrophy 0/9 0/10 4/9 Nodular Hyperplasia Hepatoma 0/9 0/10 1/9 0/9 o/io 0/9 Ductular Hyperplasia 1/9 0/10 2/9 Cholangio-Hepatoma 0/9 0/10 0/9 Hepatocellular Necrosis 0/9 0/10 0/9 Control 1/10 0/10 0/10 0/10 1/10 0/10 C/10 SCM 058187 ooo;15 WATER PCB-SD0000012727 8 Primary Liver Lesions - Aroclor 1260 Terminal Sacrifice - Rats ' Lesion * Group TI TXT Tm Vacuolar Change ' 5/25 . 6/23 10/27 Focal Hypertrophy 3/25 10/23 11/27 Nodular Hyperplasia 0/25 9/23 7/27 Hepatoma 0/25 0/23 5/27 Ductular Hyperplasia 6/25 5/23 14/27 Cholangio-Hepatoma 0/25 0/23 2/27 Hepatocellular Necrosis 4/25 2/23 6/27 Control 1/23 0/23 1/23 0/23 5/23 0/23 1/23 SC* ooor-.G WATER PCB-SD0000012728 C liolangio-lm patom a Tnlnil.iUort of Indi/idiul Liver Lesions in Knls ooo:`.;7 WATER PCB-SD0000012729 AROCLOR 1260 Tabulation of Individual Liver Lesions in Flats WATER PCB-SD0000012730 C ho la u g lo -h e p a to m a WATER PCB-SDOOOO012731 WATER PCB-SD0000012732 AROCLOR 1260 Tabulation of Individual Liver Lesions in Rats 13 WATER PCB-SD0000012733 AROCLOR 1260 Taliul.ntion of Individual Liver Lesions in Rais 14 Grading System + = minimal in severity ++ = mild in severity . +++ = moderate in severity ++++ = marked in severity P = Present, no grade SCM 058194 (no:r5~ WATER PCB-SD0000012734 THE EPPLEY INSTITUTE for RESEARCH IN CANCER October 31, 1975 Paul L. Wright, Ph.D, Monsanto Company 800 North Lindbergh Boulevard St. Louis, Missouri 63166 Dear Dr. Wright; Enclosed please find the report on Hlstopathologlcal Re-evaluation of Tissues from Female Sherman Rats Fed Arodor 1260. As mentioned in this report, there were about 20 animals with lesions, which on a morphological basis alone, could be interpreted as neoplastic. However, the definite malignant nature of these lesions cannot be proved In the presented - material because of the factors mentioned in the section entitled "discussion" In the enclosed report. . I am sorry fhr I was not able to contact you by telephone. I was very much tied uj> with several visitors last week. If you have any questions concerning the report, please contact me. Sincerely, P, Pour, M.D* Professor TMj NlWIlUUfllMlllly * MMkll SCM 058195 IM O*--y Ahm, Omtiu,Naontfca OIOS ooo;,^ WATER PCB-SD0000012735 u jxx I i i j i \ I B I T #, I I WATER PCB-SD0000012736 SncfaAtAud BI O-TEST 1610 FRONTAGE ROAO NORTHBROOK. ILLINOIS 60062 Snc. . REPORT TO . ` MONSANTO COMPANY TWO - YEAR CHRONIC ORAL TOXICITY STUDY WITH AROCLOR 1242 IN ALBINO RATS ' i HISTOPATHOLOGICAL EVALUATION OF ADDITIONAL LIVER SECTIONS . MARCH 24, 1975 " IBT NO. 641 - 96672 2 DEPOSITION ; EXHIBIT I levin-yjK SCM 0581^9 ooo:.za WATER PCB-SD0000012737 )tuLui\ial B I O T E S T 2ah*abvu*t, ina. ' REPORT TO * MONSANTO COMPANY . TWO - YEAR CHRONIC ORALTOXICITY STUDY WITH AROCLOR 1242 ' *-----IN ALBINO RATS............. -- - HISTOPATHOLOGICAL EVALUATION OF ADDITIONAL LIVER SECTIONS "IV ' ~ T_yMARCH _24, ~197S v ~ " IBT NO. 641 - 06672 ' . - L Introduction ' * ' :" ' At the request of Dr. Levinskas of the Monsanto Company, additional sections of liver from a two - year chronic oral toxicity study of Arocior 1242 in rats (I3T No. 622*07298) were processed into H & E stained sections and evaluated by light microscopy. The following report presents the results of this study. SCM 058150 WATER PCB-SD0000012738 B I O * T E S T JaixHal&uM*, Stic. 2 II. Summary In most instances, the spectrum of treatment-related histopathological findings in the liver from this re-evaluation did not differ significantly from that previously reported in our original report dated November 12, 1971. There were three hepatomas detected among three of the animals at the highest treat ment level (100 ppm) of the 24-Month Sacrifice. No hepatocellular carcinomas were observed. The other treatment-related lesions reported are regarded as degenerative or hyperplastic in nature and they are morphologic manifestations of an adaptive response of the liver associated with biotransformation of the test material. In general, the latter findings were confined primarily to test of the final sacrifice and they were dose-related in incidence and severity. In conclusion, Arodor 1242 does not appear to be carcinogenic in rats fed for two years at levels up to and including 100 ppm. , Respectfully submitted, INDUSTRIAL BIO-TEST LABORATORIES, Report Prepared and Reviewed by: D. E. Cordon, D.V.M., Ph.D. Section Head, Pathology Report Approved by: Id M. L. Keplinger^Ph.D,V Manager, Toxicology SCM 058151 goo: WATER PCB-SD0000012739 jncLuUuaL B I O - T E S T IBT No. 622-07298 Monsanto Jnc. . 3 I have examined additional sections of liver from rata of IBT No. 622-07298 and have re-evaluated them for evidence of carcinogenesis. I have tabulated my findings for Aroclor 1242 on the following pages: There is evidence of a chemical effect on the liver which is most pronounced at time of the terminal (24-Month) sacrifice. This consists of a hepatocellular alteration beginning as focal hypertrophy which progresses to nodular hyperplasia, * and in a few animals to hepatoma or cholangiohepatoma. The hypertrophic cells contain large volumes of light staining cytoplasm which is probably rich in glycogen and endoplasmic reticulum. In some sections, ring-shaped structures, probably representing whorls of proliferative endoplasmic reticulum, were seen in the cytoplasm of these cells. ' , The hyperplastic nodules occupied larger areas and often compressed surrounding cells of the normal liver parenchyma. These nodules also contained the same hypertrophic cells. Those lesions classified as hepatomas or cholangiohepatomas were larger nodules which showed evidence of confluence or some variation in cell size, shape, staining or a ductular or adenomatous pattern of growth. In the absence of metastasis, invasiveness, severe basophilia, mitoses, or other evidence of anaplasia, these are benign tumors rather than malignant tumors (carcinomas). There was no evidence of metastasis or invasiveness of these tumors in this study. SC* 58152 G00/.57 WATER PCB-SD0000012740 jiduli'Ual B I O * T E S T JaJmMiloAUt, Jxc. 4 With exception of the vacu^ar changes in the cytoplasm of hepatocytes, the more severe hepatocellular alterations were confined to 24-Month Sacrifice. * I of the CJcuial Ward R. Richter, D. V. M. ,M. S. Dipl., Am. College Vet. Path. SCM 058153 (n o:\53 WATER PCB-SD0000012741 5 Primary Liver Lesions - Aroclor 1242 3 Month Sacrifice - Rats Lesion Vacuolar Change Focal Hypertrophy Nodular Hyperplasia Hepatoma Duetolar Hyperplasia Cholangio- Hepatoma Hepatocellular Necrosis TI 0/S 0/3 0/8 0/8 0/8 0/8 1/8 i\ Croup th Tin 2/10 1/9 0/10 0/10 0/10 0/9 0/9 6/9 0/10" - 0/10 1/9 0/9 0/10 \ 0/9 Control 2/10 0/10 0/10 0/10 . 0/10 . 0/10 0/10 SCH 05815^ qoq:' so WATER PCB-SD0000012742 Primary Liver Lesions - Aroclor 1242 6 Month Sacrifice - Rats Lesion --------- ' TI Vacuolar Change 0/10 Focal Hypertrophy 0/10 Nodular Hyperplasia 0/10 Hepatoma 0/10 Ductular Hyperplasia ' 0/10 Cholangio-Hepatoma 0/10 Hepatocellular Necrosis 1/10 TH 0/8 0/8 0/8 0/8 0/8 0/8 0/8- Grout} TIH 0/3 0/9 . 0/9 0/9 0/9 0/9 0/9 . 6 Control 1/10 0/10 0/10 0/10 0/10 0/10 1/10 SCM 058155 ooor.GO WATER PCB-SD0000012743  . Primary Liver Lesions Aroclor 1242 12 Month Sacrifi e - Rats Lesion Vacuolar Change Focal Hypertrophy Nodular Hyperplasia Hepatoma, - Duetula r Hyperplasia Cholangio -Hepatoma TI 2/10 0/10 0/10 0/10 0/10 0/10 xn ' 4/10 0/10 0A10 0/10 1/10 0/10 Croup tm % 0/9 1/9 0/9 0/9 0/9 0/9 Hepatocellular Necrosis 0/10 0/10 0/10 Control 1/10 0/10 0/10 0/10 1/10 ` . 0/10 0/10 SCH 058156 \ OOO.'Gl WATER PCB-SD0000012744 Primary Liver Lesions - Aroclor 1242 Terminal Sacrifice Rats Lesion Vacuolar Change Focal Hypertrophy : Nodular Hyperplasia Hepatoma Duetular Hyperplasia Cholangio -Hepatoma Hepatocellular Necrosis TI 7/31 2/31 0/31 0/31 3/31 0/31 1/31 Croup th . Tin 8/30 . 9/20 3/30 8/20 2/30 8/20 0/30 2/20 3/30 .0/30 3/20 . * 1/20 . 1/30 0/20 Control 1/23 0/23 1/230/23 5/23 0/23 1/23 SCH 058157 00Q:\C2 WATER PCB-SD0000012745 o*-- g0 00 | || *| 4k u uu w o 4 oo -4 ao <o o> i s: s:: Ooo MO _*f8l-- 0B^I^I*4S**l*4**l *4 00<t00<Al<00<<0M<0AU0)IU')IU0I< 3 S 3 3 3 3^ o O a ' 3R -(oUJ<W'o)M'0)0M->JM| -oQ^< }0o-'<<|0>o<400>0<<0t'-<o0'l 3 3 3 3 hf 3 3 3 3 g ++ p if "5 3 tL n ___ a_ da O 3- O'OO'O'O'U'Utln 4< U U U (A <o <o <o 0<000l0<<0(00< = = = = -*.? = tp Si 8. ^ *o > 3. 3* Cytoplaomic vacuola tion of hepatocylea Focal necrosis of hepatocytes Focal lymphoid infiltrations A R O C L O R - 1242 T a b u la tio n o In d iv id u a l L iv e r L e sion s in Rata Focal hypertrophy of hepatocyles Nodular hypurplasla of hepatocyles Focal bile duct hypurplasla S'1 Hepatoma <os> Q o p ovjl CO O CJ (v--> a> Cholanglo-hepatoma teticulum Call Sarcoma metastatic Mammary tumor, metastatic Tulan(`uctauia. focal WATER_PCB-SD0000012746 AROCLOR - 1242 Tabulation of Individual Liv,r Logons in Rata ,6 Month C-I lppxn ' 10 UM 1012" 1013" 1014" 1015" 1026F 1027" 1028" 1029" 1030" c-n 1041M lOppm 1042" 1043" 1045" 1056F 1057" ' 1 1058" 1059" - c-m 107IM lOOppm.1072" 1073" 1075" 1086F .1087" J08S" 1089" 1090" "ontrol 801M 802" 803" 804" 805" 816F 817" 818" 819" 820" SCM 058159 II I ono/.c^ WATER PCB-SD0000012747 S acrifice Dose | terval Level * r rw ' . * . . ------------------------------------------_________ . .. o* ^ h1 '? 0 . 0 'SSIS'St'SSP- SSISIISSS *0-* n. 8w 0 *55388*88= * j . . . .g + + + ~+ % + . ' + ` * ' *\i KO P & . *mo na 13 g333S32asJ2 cn * > ti a S' - X a0 3 * & * + Cytoplasmic vacuolatloit of hepatocvtea Focal necrosis of hepatocytes Focal lymphoid infiltrations * ter Sr*T Do * Focal hypurtrophy of hepatocytes Nodular hyperplasia of hepatocytes F*ocal bile duct hyperplasia Hepafoma > S3 r^ h>. C o c< 3 s> r* 2 o' 'M h r> 3 ft M o au a 50 * 1 f ( 1 1 1 i 1 [ SCM ono O KH 03 r-* 0- . . '' * Cholanglo-hepatoma eticulum Coll S&rconn metastatic Mammary tumor, .metastatic 1 Tclanj'cctasia, focal WATER_PCB-SD0000012748 Tabulation of Individual Liver Lesions in Rats Sacrifice Interval *4 Month Dose [Anima Level No. and Sex oe 3o < > 2ys^i o2 * e **ne **om 23 a *uch o<Q4. 25 S' S* a. s= 5oo.o * U oAU x *oo3 fx C-I lppm 564M 566" 591" 567" .569" 572" 576" 580" 588" 589" 595" 60 IF 604" 60S" 582" 583" 607" 608" 609" 613" 621" 627" 628" 631" 634" 635" 606" 610" 620" 626" 633" + + + + ae ou W jIa>=). *0u3 f4 Liver Lesions 2; w^ e ~gs' 2<a Si 1 O 2 *v m ? 2v ea. J3 oU Cx a o 4 o x3oa o c m R eticulum C e ll S&rcom a m o ta a ta tic j 12 SCM 058161 I ooorca WATER_PCB-SD0000012749 ` to O ...... o \J\ 0rDO' N> AROCLOR - U42 T a b u la tio n of In d iv id u a l L iv e r Lesions in Rats L iv e r Lesions . " * V................................................... ' ; : ' * . .............................. ' '* 1 i S? *HdJ H{-4 B ' !t, 1 ; i .............................. a " n*1 N3S *Hn` -* Dose Level 4 O' O' O' ^4 *! *4 *4 *4 O' O' O' O' O' O' O' O' O' O' O' O' O' O' O' O' O' O' O' O' O *0 *00 00**fr*OOOO'OO'00D0000CD->4Uli^-i^-i^-Jtfwh4kO'O'OlUi NmUNN^O>4UO4UiO UiO40'UH'00'UIi^Uh<0-4N-4U1 1 1 % S 2 2. M a. ? 3 . ++ .+ + + + " Cytoplasmic vacuolation of hepatocytea Focal necroaia of : hepatocytea Focal lymphoid i infiltrations i Focal hypertrophy of | hepalocyles P +" I * +. + t P Nodular hyperplasia * of hepalocytoa 1 Focal bile duct 1 - hyperplasia Hepatoma 1% Cholanglo-hepatoma Reticulum Cell Sarcoma .* * metastatic , Mammary tumor, *d metastatic Ti: lan a* ' 1 . sia . (< >ia 1 WATER PCB-SD0000012750 Cytoplasm ic vacuolation of hepatocytes 1 Focal necrosis of hepatocytes Focal lym phoid Infiltrations 1 Focal hypertrophy of hepatocytes C lio la n g lo -h e p a to m a AIIOC LOR - 1242 Tabulation of Individual Liver Lesions in Rats Sacrifice i Interval Dose Level Anima Mo. and Sex f 1 "j 4 Month 1r. 1. 1 1 1 1 j c-ni 740M lOOppm 722" 724" +++ 744" 746" 748" ++ 768F 771" + 775" Ext" +++ 733" 765" + 766" ++ 767" 776" 773" 786" +++ 789" 793" h+++ Ext" ++ ' |1. . ( # r1. 1. Control 8M 11" 12" 21" 23" 33" 34" 35" Ext" 46F 47" 49" 51" 52" 56" 59" . 62" Liver Lesions *---- a ^a |aa. t>L 0 --S2:* 2ac. ue a-- 1 o Z -- -- a ue ua --o eC. a 2* oao fc a 1 . c. . . . .......... P + P + 'P ++ P . +++ T + P. + + P* +P P P P +' . + +' 1 U a SCM 058163 0^0 \Grt WATER PCB-SD0000012751 l> ^ n o 3C o O v_n CD r~* O -r Grading S yste m ' + = M inim *J in severity ++ * M ild in se ve rity +++ a M oderate in severity ++++ a M arked in severity P a P re s e n t, no grade .............................. '--T 1 o 51 tr ~1 : 2- % *t < *$->* nHn* Dose A n im a Level No. and Sex [control hLi m4 *O>l OO'' O4^' Ou' It Cytoplasmic vacuolation + of hepatocytes Focal necrosis of + hepalocytos Focal lymphoid Infiltrations 1 Focal hypertrophy of I hcpatocyles Nodular hyperplasia *d of hopalocytus Focal bilo duct > hyperplasia Hepatoma \ 1 Cholanglo-hepatoma L iv e r Lesions Reticulum Cell Sarcoma ' metastatic Mammary tumor, ' metastatic rP<: h ni'i*r r.i i * WATER PCB-SD0000012752 A R 0 C L 0 R - 1242 T a b u la tio n of In d iv id u a l L iv e r Lesions in Rats 9tvduJjAj&L BIO -TEST J^Jyytdjyuzi,, Snc. 1610 FRONTAG8 ROAO NOTHS0O<. ILLINOIS 60043 REPORT TO MONSANTO COMPANY TWO - YEAR CHRONIC ORAL TOXICITY STUDY WITH . AROCLOR 1254 IN ALBINO RATS HISTOPATHOLOGICAL EVALUATION OF ADDITIONAL LIVER SECTIONS MARCH 24, 1S75 IBT NO. 641 - 0667? I i 05fll65 0 o^2f70 WATER PCB-SD0000012753 ix m B I T I WATER PCB-SD0000012754 'SiiiiW r**1 **** MM NOWtAM mo+o Srr J NOcrnltOM. llUMOt* MMI ZZI UMiT TO MONSANTO COMPANY TWO TEAS CHKOmC OEAL TOOdCITT STOTT WITH AEOCLOE US4 IN ALAMO EATS H3TOPATHOLOCKAL EVALCATI ON or ADDITIONAL uva SECTIONS MAECH !. UTS IBT NO. MI . 0M72 t DEPOSITION j EXHIBIT |cevuErts44 1 oot WATER PCB-SD0000012755 ' A.* jWiilftH/ BIO -TUT ftifrtilfrliiH IM MIONTAO* *OAO NWTNMOM.IUlHOtl MMI kfaitk 14, 1971 JW 222 Gaorfa J. U*Uakai, P1D. MoowMa Conpuy 100 N. Lladbargh Bird. St. Loali, Uliiairi 6JI44 . Doar Or. Urluhti la: tBT Na. Ml*04472 Hlatopatbalof leal Evaloatlaa W AilOaul liaar Sactloaa frwa lata o< a Two . titr Chroaic Oral Taaleity Storfr ot Aroclor 1214 ___ _______ Wa aro wbnilmin harawith >w laboratory roport dated March U, 1971: prepared la coaoactlaa with the aboro arady. Vary truly yeera* J. C. Cataadra Bmidaat ..v ' ^.000 ft J WATER PCB-SD0000012756 KENOETTO MONSANTO COMPANY TWO - YEAS CHSCMC CSAL TOBOCITT moTvasaoetos um SI ALBINO EATS KBTOPATHOLOGJCAL EVALUATION OF ADDITIONAL LIVES SECTIONS MASCH IVTS ' 1ST NO. Ml - 0M72 L latiMactloa At tbs milt ot Dr. LtTiutai *f tb* Hamit Ctafur, iMUIhiI Mctioaa ot Um from t two - paar ekntlc trtl mtellT rtaAp at Araclor U?J la rata (I2T Na. 4U -07IM) war* procaaaa* lata HAS Italaa4 aactlaaa aad mhatN bp Ufbt microtcopp. Tb* hllwln rapart praaaata tba iaalu of this itarfy. WATER PCB-SD0000012757 Uni (m Ms U art tthr >lfHit illy fwn Hit ptirt--ly wywMl la v arlfiaal mpa** Ml UMokit U. IfH. Hiwiii, Mm Mm sis Mfi Ibif.MMM MM wm| atm ad M lalmat* U M U|tei MUaM 1ml 11M mm) at M Sacriflaa wUek mm mat pmilouly tyort4.;,TVa aMr llilMiS-nlilil WMi HfTWl in m*mra4~iV~ Hafaaamdwa am Vyyrytaada la aaeam ul My am naiftilult auUnatUu af u liiptln mifoan at M Urn* mmSm! a* totnuJii iibWm ri M M*t saariaL TV* laMr tamtama mm raafln*< primarily la M uiaala oi il M Kid 14 maatk aaarifteaa ul My mm Im-nhal la larUaar* id aamrity. la rmalaalBa. Araalar 1234 iffiin ta Va tU^kHy amrl|alt at Unit at 100 ppaa nVaa M caadaoaaty la M Dm far tm ymm. Raapscdally nkaind, QCDCSTUAL UO-TOT LABGftATOUES, INC. WATER PCB-SD0000012758 BIO-TItT ibt k*. tu-mn ] 225 I Wn ohM iffttl--1 rntUn rt Uir tna nM rt ttmtr Wurbir tU-71W amt harm w*mlHi4 >Mi to* wlfciw f MrtmiMiU. tbm t*k*ltof mf flatftog* tar AimIn US4 *a ikt tottovlag m. TWn to aritramr ri t tkMtal tBM *a to* ttm <Uck to a**t gronni it ttoM at to* ^-Um* u U*UMk wcriOc*. Tkto cm*1m* of fc*p*lnr*H*lr iltmUH h|l--In m toe*1 h;g*M*fby *tkk pnfniM* to tolir byparpteato. amt to tow utatli t* b*p*taa* *r rh*l*gt*b*p*toTn. Tkt b|yili*>Mi e*U* ftool* l**g* *n^ of light miatog CTtaplaam *Ucb la prok ilf rick to gtfeaga* uf nfepUimlt rorlrohw I* mb* -- iii| h*p*f iimBu*. pfhlly r*p*****Ctog k*(li rt pnlliintiM atofUamlc ntlnhaa, wu* ***a to to* cftopl**m of to*** mIU. TIm bypaiplaatto --<*!** *c^4f larger ***** **f *ftoa cmpniMf --------"-g call* *f to* Miaal Unr pmockrn*. TWy *U* c*at*to*4 to* am* >ip*ili*pMi call*. Tfc*** toatoa* wUck I claaaiftof *a hoytown or ctoUafU-toytMou wm larg** i*Ma* which *h*w* f *t!4*-- of e--fto*f* * mb* tuiulM la call *la*. shag*. **totog fwtoti i4***b*imi **tt*rm rt growth. I* to* k*--* rt **aataato, l*t**lin*i*, nmn kanffclU*. town*. *r to*r lilmi rt --r``-'I I cIwm to *U to*** baatya toner* ntooi too* ailigM*: (carciaraa*). That* w*o a* ntfmM of ii*i*ili * Imalnati* rt to*** WATER PCB-SD0000012759 tIO-TIST 4 tZ 6 Wttfc ULipMii oi tfc* 44C--U* ckMf*a im tfca nn>hw *t 1 Mti Nnn Wf f--Malar tUmlltM *t caafla*4 M lateb f ifca 14 > SmtUIm. War* ft. fticfctar, D.T.M..U.S. MfL, Am. CaUafa Tat. PMk. OQO'TS l WATER PCB-SD0000012760 227 Primary Lhw Uiint Amlw llS4 ) Mmk OaarMWa lilt l' Urt-- . ant rt TS TZS ^w Camral vtcttUi 3/10 3/10 t/io 4KU 1/10 reil If>yaUoyhy WIiIit RyyaryUaU 0/10 0/10 0/10 0/10 B^mma Dactalar HyptryUila 0/10 0/10 0/10 0/10 Chalaafto -Hayatama Hayatocallolar Nacraala 1/10 1/10 0/10 0/10 000 0V>4>*/> WATER PCB-SD0000012761 *-ly 4 '* i ' tC- v Primary Um Laai--a - Amlar 12M 1 '* " UbMk ImiUIm JUt 3^. - Laaioa Vuwlu Cktaft Focal Hypertrophy Nodalar Hyparplaria Hepatoma Dwnlat Hyperplaaia Qtolaafio- Hepatoma Hepatocellular HcerooU TI 0/10 0/10 0/10 0/10 0/10 0/10 Z/10 Tn 0/10 0/10 0/10 0/10 0/10 0/10 0/10 vm v 1/10 * Cmnl 1/10 4/10 K. 0/10 0/10 0/10 0/10 0/10 0/10 0/10 0/10 0/10 1/10 1/10 WATER PCB-SD0000012762  Pilury live* Uia - irocUr 1254 14 Marth OMrlOea - UM Lio V Of*- a in TO *>?. CMW YumUi CVh 1/10 4/10 1/10 e/ 1/10 r*c*l Hipiniyt) 0/10 4/10 2/10 *Ao 0/10 NotaJ&r HrparpluU 0/ld 0/10 0/10 0/10 Maptoot 0/10 0/10 0/10 0/10 tecttlu Hyparpteala 1/10 1/10 0/10 1/10 Ckolu|i'*H>P*B* 0/10 0/10 0/10 0/10 Hptoc*UaL*r HvcrorLa 0/10 0/10 1/10 o/yo 0/1C T * a WATER PCB-SD0000012763 230 Llm tirtm * Amtor 1254 Tarmiafcl ImtISm * tut LU VtnalM Cku| rocAl HrixrtTopAty Ntdalii Byp*rfUl HtpMBt Doctnlar Hyprpl*l Cfcotoflo -HpMWM HftncUuUr Krfi * *** |_.-y _ar 9 **~-- 1 * $4* ila.t /3 3 1 Ijfc i* i T1 * Til * p 4 e/ii iA Vd 10/25 -/ia"/. X.H 3/31 y>ifa S/24 y*r*.v 13/27 Vn*\|J/2J 0/31 % 4/25 13/27 1/23 0/31 0/24 V 4/27 >--i^l e/23 0/31 vv vd 3/24 v.V 4/27 5/23 0/31 0/24 3/31 V* v*4 1/24 1> w * ` t * >L ^f >t * tr 2/27 */ir ^ 5/23 2/27 >i-|1/23 k, "'-T r^. *--r WATER PCB-SD0000012764 WATER PCB-SD0000012765 AJtOCLOS . 1214 TiMMIm W M1>UmI Urn UiUm la Bala m 10 'r-r _,. *Tt \!rt ooo;.c2 & WATER_PCB-SD0000012766 ,1 WATER PCB-SD0000012767 TP WATER PCB-SD0000012768 arocu* im is WATER_PCB-SD0000012769 AltOCLOR - MM TtWUtlM af ladlvMaal Unr UlUu la IUU 236 3*c rifle* b^arrml Dom lAaimal Laval No. aad s 3 it !iSaa ll tl si J fa IV*a Uw Laalaaa ii J a T XU3o ii i 24 Month B-m lOOppm * / ' A*-4 r4I4M 491" 497" 499" 507" 909" 110" 912" 4*3" E409" 9UT tf1 H 924" 927" 941" IT347" Eitl Ext2 Cat3 934" 337" 6\,i 339" 441" Xzl" Cz2" m+* Cx3" iw (** fcr 8; H PvH P l * iW t'* if> i* 9* iI* # K*19 ^ v,r p *n WP M ipi p Pl-> t H P"8'-: ft'-1! p p id** (f'jH I ItII * If't ** w> ** Jw P i-lj h a-! B Pi IH* M iiitm iry turner tu tiu U llv 14 C ra<lin Sratam * --<<-< |a ta rarity *- mild la aararity -v* modanaa la aavarlty hh nutBad la aararity P Praaaat. aa ptda Abaaal . WATER PCB-SD0000012770 U-Juai tIO-TIST 2 237 11. Summary ' t -'- A In roost initaacasj, treatment-related histops thole. .1 fladiags la tbs livsr from this re-ewlnatiae 414 cot 4illsr slcniilcs; (root that pntioulT reportsd In our e Ifins1 rsport dated NoTomhsr 12, 1971. However. the-a wars six bsalfa Liver tuxuora datsctsd smoaf six of tha animals st tbs highest trsstxnsat IfTO l 1104 ppm) of tha 24-Month Sacrlfica which war* not previously - -- .e ... . . r( '* . . - reported. Tha othar treatment-related Issloas reported ara regarded as ` degenerative or hyperplastic la nature acd they ara morphologic manifestations of an adapeiva raspoasa of tha liver assnrialad wish biotraaaforroatioa of tha tast rratarial. Tha lattar lasioas wara confined primarily to last animals of tha 14 and 24 month sacrifices and thay warn dose-related in incidence and severity. la conclusion, A roc lor 12S4 appears to ba allfhtly twnorigemc at levels of 100 ppm wlian fad contiaously La tha diat for taro years. Respectfully submitted, INDUSTRIAL BIO-TEST LABORATORIES, INC. Report Prepared sad Reviewed by: 0^3 D. E. Cordon. D. V.M.,Ph.D. Section Head. Pathology Report Approved by: M. L. Kepltybr. Manager, Toxica! WATER PCB-SD0000012771 JMiatud aio-risr AkrAvu, 3*. 1610 NONTMI frOAO - _ NO*Tn*kOe.-t.llM01 M***= . 188 REPORT TO MONSANTO COMPANT TWO - TEAR CHRONIC ORAL TOXICITY" STTOT WITH AROCLOR 1254 IN ALBINO RATS HISTOPATHOLOGICA L EVALUATION or AOOITICNAL LIVER SECTIONS MARCH 24. 1475 1ST NO. 64. 06672 /' WATER PCB-SD0000012772 ItM nOMTM NOtTMMOOt, illlMOIl jm. 189 Cni J. Lortaakaa, ftJ). MO N. .'.dbw| Bird. St. Uiui, MUiwrt U1M Dost Dr. Lovtaskos: IU: BT Ns. M1*4UR - Ill--npitWilmtcal tnhulai at AMittaui Uvsr lirttmi Fran Xm gt 11 - Yatr Chfnic Oral Tosictty Studr ai Amine UM. Ws sro iihin|g| hvnrttt oar nvutd laboratory rap art dated March 24. WH; prepared ia rmairtim with Oa abev* study. Vary truly yours. i. C. <~lBdr WATER PCB-SD0000012773 * V - '***&>: > .,y..**'v- " ^ --T.nn^-a-A'.C. 5:' -Li;-L BIO.TIST \ i. REPORT TO MONSANTO COMPANT TWO TEAS CHRONIC ORAL TOXICITT STUDT WITH AROCLOR U54 IN ALBINO RATS KBTOPATHOLOGICAL EVALUATION OT ADDITIONAL LIVER SECTIONS MARCH K 167S IBT NO. Ml. 06472 L IntTodactloa At taa n^Mtt of Dr. Uvtiilai of tkt Uuiutt Cinfur. iMiiiohI Hctteu of Unr Iron t ear* - yoar ckratic oral tesicity iMy of Aroclor 1254 la rata (IBT No. 6LI-07ZT4) wore praeossod laio H 6 E itaiaaO aactioas ud svaluatod by ILffat microscopy. Tbo !tllnu| rtpor pro .oats As roaulta ol this study. WATER PCB-SD0000012774 *++td BIO-TIST 191 > . , tfr . n. la ia tha i ad m NltMkM raportad la oar m Iglanl I six hailgo liror 1 Wml C1M ppm) od tha H Halt 5arr*ta tshteh oui aat prarlanaly rapartad. Ou livor tumor <a hopoaama) prortsuaty r^orOi ia T-0 animal (Ho. 445) ni n-diaUM as aodola? kffiyinli Tho athar truialralalid laka raportad an Mfirdad aa dagaaarmttvo ar lifpmplialli la aaama md thoy art morphologic manlfaalalliaia od aa arlapHra roapeauo od tha tivor aacrlbad ta biMnuigraattaa ad tha boot notarial. Tha Umar laoiaaa oaaa ranflood primarily to toot animals od tha II sad 24 month aarrlflraa aad thoy U1 ifciaa ralitail *n inrlrUnro aad sorority. la conclusion. Arodor 1254 appears a bo slightly nimorlgmUc at larods od 104 ppm *boa (ad nattaooaly ia Aa dlat iar too yaara. INDUSTRIAL 1D-TT5T LABORATORIES, INC. Report Proparod aad Roviowod by: D. I. Cardan. D.V.M., Ph D. goctlan Hoad. Pathology Rrpor i Approrod by: !d U. L. KapUngor^Ph.D.7 ^------------Tm-l . ' WATER PCB-SD0000012775 t * 1 BIO.TUT c ibt n. ui-mn 192 I ktff mmliri additional oactioaa od lifu (ram rata ad Jndr Moslr 422-0T29# aad bare re-araluated than lor arldaaca ad carciaageateti. I bin tabulated my fladtage for Arocler 12S4 aa tha loUawiag pages. There la arldaaca el a chemical affaet aa tha User which ia moat pro- aeaacad at rima si tha 12 -Month aad 24-Month sacrifice. This constats of a hepatocellular alteration beginning aa local hypertrophy which progrennea :a aadalar hyperplasia, aad la a law aaimali to bapatoma or cholaagiohepatoma. Tha hypertrophic colli contain largo imonnta of light staining cytoplaam which la probably rich la glyeogaa aad aadeplaamlc rtticnhim. la aama caaaa r-.sg ahapad atroctaraa, probably rapraaaatlag wharla a1 pretifarntire aadoylainuc reticulum. wore aaan la tha cytoplaam o< tha aa calla. Tha hyparplaatic nodule a occupied largor araaa aad often compressed surrounding calla ol tha normal !' s* ctraachyma. They alto contained tha same hypertrophic ealla. Thoeo laaloaa which 1 claaaUlad aa hapatomaa or choUngio-hepatctrna ware larger nodule# which ahowad arldaaca el ceafluaaca or aama rarianon ia call also, ahapa, itaiaiag or a ductular or adiaamatoua pattara ol g.-oarta. is tha abeaace al mataataala, laraslreaesa, tarere basophilia. mitoses. ar other arldaaca ol aaaplaala; t ehaaa to call thate baalgu tamer a rather than maligna:: (carcinomas). Tharo was so arldaaca al matastesia ar laresirwassa al theta tumors la this study. . -- WATER PCB-SD0000012776 mor* MTirt h*p*mc*ltal*r tltintloM war* rnaflaii la ulnb af At 2411 nmth Sac rifle*. War* It. licittr, O.V.U..U.S. DlyL, Am. CaOaga Vat. Path. i1; `utrwr ' (*A*r* 1 . *2TjL-, . r.JU.'Mfc 7 -,t vi> i- w 000:33+ WATER PCB-SD0000012777 ... -a . - ...ic - 1 M^3iK: v s . -s^er.;5>s^M|^: '-* -.- . V ' 'V'<`.VV'Y^'* V't.* - JR** 'fZMtZix WW j\Jt VwUt CUa Fool lljpaimiphT No4*Ur Hypai-plMte DkmIu1 HyparpUjia ChoUayio -Hapatoma Hapatocaltalar XacraiU ***> ';$.-sr-T?* *3v . >MiN n/w i/xo o/ii 1/10 0/M 0/10 0/10 0/10 0/10 0/10 0/10 0/10 0/10 0/10 0/10 0/10 1/10 1/10 0/10 2/10 0/10 0/10 0/10 0/10 0/10 0/10 r.-*V*.> ... f _ v[1^/^ A' ,' : n* 000134 DiliHiiMBa WATER PCB-SD0000012778  195 t Primary Um Udou - Aroeler U54 1 Maatfc Sacrifice JUt* Vacuolar Cheap* focal Hypertrophy Nodular Hyp* rp1**1* Hop*tom* Ductular Hyperplada ChoUnpIo-Hepatoma Hepatocellular Necroat* TI 0/10 0/10 0/10 0/10 0/10 0/10 2/10 in 0/10 0/10 0/10 0/10 0/10 0/10 0/10 TSX 1/10 4/10 0/10 0/10 0/10 0/10 1/10 Ceatrel 1/10 0/10 0/10 0- 10 o/:a O'lO 1/10 WATER PCB-SD0000012779 7 196 i Primary Liror Lotloaa Aroclor 1254 U Uostfc hcilAct Itu Loolon Croup a th Tm Control Vmglu CUa|* 1/10 4/10 1/10 1/10 Tseal Hypertrophy Nodular Hypo rplasla 0/10 o/io 4/10 0/10 2/10 0/10 0/10 0/10 Hepatoma 0/10 0/10 0/10 0/10 Duetula r Hyporplaria 1/10 1/10 0/10 1/10 Cbolaaglo- Hepatoma 0/10 0/10 0/10 0/10 Hepatocellular Necroaia 0/10 0/10 1/10 0/10 V. WATER PCB-SD0000012780 H7 Primary Lhtr Larioar - Aroclor 1254 Tarmiaal Sacrtflca Rau L*ties VtcooLar T'jcal Hypertrophy >Vo4ular Hyparplaaia Htpuooa Saucular Hyparplaaia Cholanfio-Hepatoma Kap4tocitulAr Ncfasl< n 8/31 1/31 0/11 0/11 6/31 0/31 3/11 Til 10/26 3/26 3/26 0/26 3/26 . 0/26 1/26 Grog* TIB 13/27 13/27 11/27 4/27 4/27 2/27 2/27 Ce. 1/23 3/23 1.23 0/23 5/21 0-23 1/23 WATER PCB-SD0000012781 AROCLOR UM TttatUtlw of WMtal Unr Ullou la Rata Sacrifice lac* ml ) Niooth Deaa Laval 4 aim* No. l and m nSaa a It ll ?s t si Si i* SI 1V 9 g U X u B-I |1 33 4M + 337" 351" 33 9" 9* "1340 39tr 397" ! 393" 399" 400" B-n lOppm 41IM 437" 433" 439" 440" 476T 477" 473" U79 480" 'i fi ! H t Uaar Latioua ij 9 m H 1 . 5J V * j1 i* 9 * la 1 I j I . ' i i1 i i ij t ii I i 1 i ; i i j r 9 J i iII ii i I B - El 516M lOOppm.317" 511" "1519 I9i0" |ssr !S37" 331" 339" | 5o0*' l *- | i fj i i j 1 ij \ i .t -i i i i 198 ii s s =l . K 3c Zi 12 3 Z it jl p i. ;i j' i i i i i I ) WATER_PCB-SD0000012782 Oo U Z * Oo o Vo -o ->o1 p um m^ mw m O -4> to `I ^ W N I it * . ISO ie : : 3" i 6 M o n th ' B - I 92t M { || : - I 1 Ippm ** to A 2: if s * MO.. |Tyioplanilc vacuuUllon of hapatocytaa Focal Mcroait of hapAlocytai Focal lymphoid inflltraliona Focal fcyptrlrophy of iMpalocylaa Nodular Kyparplaaia of Uplocylaa Focal Mia duel byptrpfaaU lUpaioma is U ES 21 ra rn i S' 9m u v Cholaoglo * Impatoma S|uamoua Coll Card nomamuIndalle Mammary lumor. malaatallc /('Irrlmala to 1G # WATER PCB-SD0000012783 AROCLOR - US4 TtteUliM ol ti41Ul Unr UiioM la Rata fertile* latarval Doan Laval fcnlma uNo, *&4 S** | 2a Sg ll si u If*s *l 11 -3 f fraa 4U U. 8 u u Uvar Laalooa If If it ii i ii8 w 0 Z i& fi 4 8 w i jj U Month B-I lppm 1 41M 42" 43*' 4-*" 4S** 4r 47" 48" 49" 50" j B-Q lOppm ' SIM 52" 53" 54" 55" 5&r 57" 58" 5 9" 60" 4+ + * * * B-tn blM 100ppm ' 2" 63" 64" 65" +* ur 1 67" 68" j 69" 70" ' i i `r > 11 200 % *y- ffiw&.a-#SrSSE t ? 1 000200 WATER_PCB-SD0000012784 3crlflco Intom l AnOCLOR 1234 TihnUtioa of laltoi4tal Unr Laol ji la Rato Don Lvl |Aaima( No. and Sox }\ u sl ! 1? STS ft 11 Llrmr L^ttpno * it iL 1 1 2 2r u1 201 - + Month I B-t 326M< Ippm ! 310" 333" 338" 331" 123" 323" 343" 144" 342r | 342" I 341" j 374" 173" 176" 473" 366" 367" 372" 177" 378" 374" 360" 381" 384" : 187" ! 388" j 38" J 301" I 394" !' Cn" i Il u WATER PCB-SD0000012785 M a iiu n A ry turner mvldiioitir AROCLOR 1214 TtkiUHos o( Individual Unr LtilM la Rata Sacrt/tca Don Ifcaima Intarval Laval JNo. and Sax 3 m Si, $1: -1 |j F I KI u-- 24 Month B-n 4 ISM lOppm 417" 4 IS" I 411" j 423" j 42b" j 427" ] 491" 402" | 401" I 405" i 410" j 414" 14Sor I 411" 1452" | 45b" ! 459" | 4bl" ! 4b9" ; 470" 471" 473" : 44!" '445" LIvar Laalaaa n2 i a I1s= c1 11 1 fr K H n ii*3 i z* P P t * ?a J U 102 11 , At' ' -- il W000202 ** nd. WATER_PCB-SD0000012786 AKOCUm 144 TlbaUllM ot ladtrtdoal Lirar Uiioai La Rtli u 203 Sacrifica latarral Data Aalma Lirar Laaloaa itLarat iNo. naad a Ju ?i Saa a ** U jl * fi is U it 11 m3 3 -f2 1 S- i V1 y ia h. iSk i! 8 i ( 1 21?i 4 2u i -:i 5- 5: ea iiIsf i1i Hi! 13 n 24 Month s-nx lOOypan 444M 491" 49?" 499" 107" 509" 510" 512" 441" 449" S2iT 524" 52?" 541" 54?" 44 4 44 4 4* 44** 4* **Extl'l Cat!" ' Ext4" . tid" 516" 517" 519" 4 441" Cal" Cx2" 44 Cal" 4P 4* P 4P P 4 * * 4 4*44 *** -* *4 1 P P P P P P P P 1 * * - P* p ` 1 p ? p 1 p i p 1 1 .~ p Grading irtaa . > minimal la laranry *- mild la ararity modarmta la ararity marfead la ararity P PriHM, as grad* Abtaat ~*MSnteL* .mt&d v't.r/- 000203 i- WATER PCB-SD0000012787  * . ->V/ ife ..v.. f\\ -A 'is*.*".. .> ; V Ay ' :-: ^ . . i.. ; w ^v"' > - \. w*k jUMA^ItO.TIIT NOAfwMO--. UUWII NMI v - II - ,,, .0 DtfQlltO MONSANTO COMPANY TWO YSA1 OttOMJC ORAL TOXxetTY STODY WIT* AI06LN M IN ALA040 IATS . KtfTOPATKOLOOtCAL (VALUATION or ADDITIONAL UTOt IKTIOM MABCK 14. lf?l t>T NO. Ml sun FI 1 : 9 4 WATER PCB-SD0000012788 Bai arm. Hi-*un - mx^irtnimini u Ltnr BacHaaa fna Ui af a Twm - Tear rtewtf Oval Taaldty < Avadar 1W4.----------------------------------------------------- Va a* ibattia| bawllli mm f ariaad laboralary i^ial da*ad March M. 1TTV yr^arrd ta eaaaacttaa whh ba aharc atatfy. ymj ndy yaura. t. G>mje+~mU. 3. C. Calaadra JCC/U WATER PCB-SD0000012789 4 WiUrf ilO*TI$T -',058 (XPCtT TO MONSANTO COMPACT TWO - TEAS CXBCNK OftAL TQXICITT STUDY WITH ABOCLOB USA IN ALBINO BATS H3TOPATHOLOGJCAL CVALCAT1CN or additional litzb sections VUICH 14. in 3T NO. Ml - 044T2 L at24actlaap At tfca raqorat a( Dr. Lavtaakaa al tha UntuM Cssspaar. tAditiseal actloaa of Um (ram a tv* paar cknalc aral Mrtcliy itt^y at Araclar 1254 - la rata (IBT N*. 622>97290 vara pracaaaad lata Hit mM mc&obj ut rralaatad by Uykt microacopp. At fallcwlaf rapart praaaata Am raaalta it tfala tto4r> WATER PCB-SD0000012790 iraaHnal late (lM pyn) H te llnadi SaertXca. Ha k^ima.ItaW cardaaaaa vara atemW. OH Inr toar (a lipiliai) fnrlaity roporto d Is a T-0 rr1--' CNa. HS) bm ta-rloaaHlad aa aodalar hyyorplaata Tke hyparylaaftc Is iiHi ate ter ara anrpholnylc inlfiiHHn H aa adopt!** raapaaaa at te Haar aaaibal H btotraoabnattaa at te Hat notarial. Tha c * LaeHr Iteaoa *ara iimflaTi prlsailly H Hat oataala at te U ote >4 ho4 oacrtXaao ate tey ** iota raloail Is larlilaaca ate aararlty. Is aaactetaa. Ante 12M appoora aha akffifcy'naatiaak at lorraU I'Xm, v-'c ,'-w* u< * * . .. _. at 1M pys ates lad sateaclf la te dUt lor Ih yaara. , MDUSTX1AL BO-TXST LUONATOUXS. INC. Xaport Praparnd ate Irrtate hyi D. X. Qardaa. D. V.U., HuD. Saatiaa Hard. Patetafy Xapart Afpratad Vyi WATER PCB-SD0000012791 WATER PCB-SD0000012792 WATER PCB-SD0000012793 WATER PCB-SD0000012794 4 083 Primary Unr Uittii - Aroclor US4 4 Meath SacrlXlce - Kata Lee ion Vacuolar Chaaye Fecal Hypertrophy Nodular Hyperplaaia Hepatoma Duetolar Hyperplaaia Cholaaflo-Hepatoma Hepatocellular Necroale TI 0/10 0/10 0/10 0/10 0/10 0/10 2/10 TB 0/10 0/10 O'lO 0/10 0/10 O'lO 0/10 Cl Tin 1/10 4/10 0>10 0/10 0/10 0/10 i/io Control 1/10 0/10 0/10 0/10 0/10 0/10 1/10 i WATER PCB-SD0000012795 PitaMT U*wr Uitaa - Atmlor 1254 U Most0 Ontlfln - uu laiis* Ytralu Cta| Foul Hyyrtmyhy Nodultr Hyftrfluli Rapatcma Dctl>i HyprpUil* CboUaio-Hapotoma IbpwoMlhUr NtertiU S 1/10 0/10 0/1O 0/10 1/10 0/10 0/10 TO 4/19 4/10 0/10 0/10 1/10 0/10 0/10 nn. 1/10 2/10 0/10 0/10 0/10 0/10 1/10 Cooftrol 1/10 0/10 0/10 0/10 ' 1/10 0/10 0/10 WATER PCB-SD0000012796 5 A;- *? / - ii ifTThrV i** .;ijy ^., v, /. iif iii...i ------ MS A> '. c Tifcai) Urn Urtw m* Tiwrlr-* hotflc* ftatt.g&i 0/23 MataUr im 1/20 13/2T 1/23 BafUan* DwcIsUr Hrp>UU 0/11 tm 0/20 3/20 4/2T 4/27 0/23 3/23 ( ChoUatto-HipO-- Hapttocallate* Bril* 0/31 S/31 . 0/20 1/20 2/27 1 ~X/27 0/23 1/23 WATER PCB-SD0000012797 v.* fa. !i X fl H fii| |CflofUinic vicmUUm ol Wapatocyf fw>l (MrMll < hlfilKflM Focal IfnifteM wninuaM Focal Oypartrf by of hopotocytoo IMaUr kffaiflaila a! hapatocytaa Focal Mia 4ocl byparplaala Ikfaloma t, H4 f * l> is h i* rf I W ts P Cfcolamlofciy< am' looniwi Coll ' Carcloaoaa. oaotaatMIc Mammary locnOr, malaalalla nipi!ilia /Cirrhoala' WATER PCB-SD0000012798 ABOCLOB . 1*34 Tabalatiaa ot Mhrtdual Lint Ltliau la Kata SacrLOca Lntarval Daaa Anima Laval No. aad Sw II si f**L lf Si f3 m l U-- B-t lrpm mu 9Z2" 9*3" 9*4" 9*3" 934T 9JT" 938" 34** 940" Uwr Laatom it v H a i! L 1 2 V 2 2u I B-a MOM j lOppm 931" 93*" 933" 954" 944T 947" 943" 949" 970" B-m 93134 | lOOppml 932" 983" 934" 933" 794T 998" 999" 11000" 067 2 2 i! 55 iM 10 IX It a 29 ! <3 WATER_PCB-SD0000012799 r fa | x * 1 tt t 4 ' i ` C ifl f ? f" II * Focal kwrMil of . Iwyaaarylaa Focal liiijliM loftlttiM-- r, * Focol fcyycMl^fcr rf fcofo*oay' a - ** Na4ala*.tifparpla a la at lofltMflM Focol MIc Art! * r kryciyltila .. f -J.fSV:, V ChoUotUliii|Cilocn* ' (luucm-- Coll ' Cm rclooma moteatatW Mjnimtri timor, ingUfUtic lk|i4tllls/(!(rrlioil J fey . : jM > .f 15 ' .. '.**iV WATER PCB-SD0000012800 T ik iU d M o f UAiRr MOCnLtOl RL i-n ii K 0 1 *- ' fs r 3: MUWWWU0WW0WWWWU*UWWW00WWWWWUWWW Q^m<0 0>4J.m04(|N*49>UI9>UI^)NN9'W<I>WmuiOO' zz -.*t z z z z -X i* 4 +1 1 : t ** n* 1i Cytoplaatrlc *ani*ltUl of U/ilDcytsi;^:''' Focal naaroala of UfaiasyUi' Focal lympfcol^ ...jj-w laflltratlaaa "j t t t t Tocol byportvayty .of hapatacytaa ^ Nodular kyparpliala ji of hapatocytaa f *1 Focal Mta r hyparpUaU^^. S' e lUpatoni* * .i Cbolaaflo-bapoMMM* ,V - ' Bqucmotu Colt '- Ca retail ma.mataialaUc . .. Mammary tama*i .. molaataCs --------------------------------------------------------------------------------------------------------------- -------------- -----------------------------------llapatltla/CIrrhoala I Ut M La i le i i la Rats WATER PCB-SD0000012801  *. . i ^ . -n . it MhartfrAfiin ii rfV*fc<S . -*\-. j ,, AAOCLOR - US4 T&baUtioa of Mlvtdaal Ltoor Lotions la JUu Sicrlilc. Interval Doi. Kn'ja* L.v.1 No. Uw UlleM V- 24 Uoath Su Ii ]i li 21 Si i0* iL_ B-B 413M lOppm 41?" 413 419" 1 423" { 426" ! 42?" 491" 402" 403" 403" 410" 434" 430r 431" 432" 436" 439" 443" 449" 470" 471 47 443 443 ii u >i H j i ii 11 11 i P P i j ; ) i 070 u a i Ii is: ri e S S -I WATER PCB-SD0000012802 AHOCLOR i&ii m r<i 4* h; V n Al * t 3JC o 5 n p fr m f ..... .. . ....P1 ..................... f? 3: PPPtSSSPPPPPSSSSSSiSSSSSSSfc 8 n? 0 4- 14 4 f t tt tM* + 4t > t Cytoplasmic vacootallon of hptocyti N r a Focal oacroala of (T hapalocylaa 4 {It t ti * 4 + Focal lymphoid Inflllrallona Focal hypertrophy of hapalocylos ft I I o O'O'O du'a >o >o d *o *d *a n <d Nodalar hyparplaaia of hapalocylaa Focal hlla duel hyparplaaia llapaloma r 3 r o ff 0 S' V a .. -- -- . - -- --- --~ ---- -- - -- .Till '" * < ""y C hoUg16-hopaloma Squarooaa Call Carciaoma, mala alalia Mammary lamar, mataatallc lli jMlilii/Cirriioali 4* WATER PCB-SD0000012803 Laaioo Vacuolar Change Focal Hypertrophy Nodular Hyperplasia Hepatoma Ouctular Hyperplasia C Solangto-Hepatoma 1 ia pa toesHula r Nacrosia o 072 Primary Uni Uriou - Aroclor 1254 Tai mill Sacrifice - Kata n 1? 1 1/11 S/2* 8/11 TO 12 Gray TD1 1 12 1 4/26 4/25 10/26 11/27* 1/2511/27 I/ll 2/2S 1/11 0/20 0/11 0/20 0/11 l/ll 6/20 6/11 4/2* 0/11 2/11 2/2* 1/11 1/26 1/2S 5/26 1/26 1/2S 1/26 0/25 0/26 1/26 4/25 1/26 0/2S 0/26 1/26 2/25 1/26 6/27 1/2111/27 1/27 9/2511/27 1/27 2/25 4/27 5/27 2/25 4/27 0/27 1/25 2/27 1/27 1/25 2/27 Control 12 1/21 0/24 0/23 0/24 1/21 0/24 0/21 0/24 5/21 6/24 0/21 0/24 1/21 0/24 1 - First anlutioa af arightsl slides 2 - Ra valuation ci oHyiai 1 elldea 1 - Evaluation at artdlrtnaal tiaauo aacHaaa Thera vara $ aaintala ia which tMa laaioo was not aaaa at lima of Iha aacnad avahaettan hat was praam on lha first. 1 Similar to above Involving 4 lalaala. ` -- u. WATER PCB-SD0000012804 If WATER_PCB-SDOOOO012805 WATER PCB-SD0000012806 WATER PCB-SD0000012807 WATER PCB-SD0000012808 t l E> xi H I! B| II WATER PCB-SD0000012809 ZMO IM 'OK III iU>\ >* H3VVTI SKOtxoas uati ivNoiuaav jto N0UYQ1VA3 TVDIOOlOHiVdOiSm 61V* OMglV MX <** rm HOTDOHV Hi i* xanx* A1I3IXOX 1VVO OINOVHO VV3JL OM_L XNVdnOO OXMVSNOVf ox X80aa (MH iomiih ieo<iMaoN oz are* crin m*i 'mqnivf xsix-ott 7"*r*f% * WATER PCB-SD0000012810 IhluilW io-tiit Jabnviknm. Snc. *10 OHTAOI O*0 MMTMBOOK.ILUKOi> ZOb Gsor(s J. LaTiBaha.it Ph. D. Monsanto Company 00 N. Udbor|h Bird. St Loots, Missouri i)lM Dsar Dr. LaTiaakas: Bai IBT No. 441-06672 Hlatopatholo|lcal Evaluation o( Additional LiTor Suctions From Bata uf a Two3 Tsar Chronic Oral Toaieity Stn^_^iiAroelor_W4A;_i_v^^ __ .I Wo an submitting harosrith our1 laboratory roport datsd March 24, 1975. ;*oparod 1b coaooctioB with tbs abovu study. Vary troly pin, J. C. CoUadra Pmldtit WATER PCB-SD0000012811 UM BIO-T3ST 206 REPORT TO MONSANTO COUPANT TWO - TEAK CKXCtnC ORAL TOWCITT STUDT WITH AROCLOR IMS < U.0 IN ALBINO RATS KBTOFATHOLOGICAL EVALUATION Or ADDITIONAL LIVER SECTIONS MARCH 24, 147} DT NO. 641 06672 L Introduction At tha rv^wct of Dr. Lavinakaa of tha Hoatute ComfUj, additional iite sactlooi of lint (root a two yur chronic oral toxicity study of Aroclor IM to rat* (1ST No. 622-07291) an yroca;id into H 6 E stainad sections and amlnatc-d by light microscopy. Tba following report presents the results of this study. IO.TBSV 2. 207 n. Klim Mlljliriltl rt IWU|i Id ki Um tna this w-<nhtrtw 414 an* diffar ripBcuUr Imm 4ii pwrimuly r*pcn*4 la oar *tl|tail npnt dial Hmirdli 12, 1971. Ihwm. than ware m*w fcaolpn Urtr tunora datoetad invwa aoaos W 4i nlaili at tka hl|ha*t tron&ao* Wool (100 ppm) of tka 20>Maotk SaerlAca vktck vara oo* provtonaly raportad. Tka otkaT tmOmit-nUad laaloaa raportad ara rtfiidid > d|ari diva or kyporpUatlc la aataro aa4 thay ara a**rpkoloflc maalfaatatloai of an adapttfi roapoaao o( 4a livar aiaociatad witk biotmmafonnotloa if tha taat material. la oaaral, tka lattar finding* vara fiaflnad primarily to taat animal* of tkaJ*. 14'- 24>Moatk Sacrlilca*. and tkay vara doaa*ralated In Ineldanc* and sorority. Hi la eoadualoa, Aroclor >211 appaar* ta ba allfktly tumarifanic at Laval* of 100 ppm vkaa fad caetiaaooaly la tka 41at far rva yaar*. Roapoctfully ankmittad. INDUSTRIAL BIO-TEST LABORATORIES. TNC. Boport Praparad and Ravtawad by: D. E. Cardan. 0. V.M.. Ph. D. Sactiom Hood, Patkalady ?or' V WATER PCB-SD0000012813 208 I ten tnmlMl itfUoMl -- cHooa of Um fan rate l( ST No. 411*07211 Hi ten r*>*ilsaMi iteot for svidaaaa of anrdaapoaosU. I ten Utelatei oty flodtog for Aroclor - i *& aa th* fattening fip~~ Ttert it tMum of o rhomlral ofloet oo teo llnr which n> mart '1. ' evident at time of tteririiMk oof terminal (H-Mtek) aacrtfico*. This conoid* of o fcapatocaUdar nitaratio* haglnntag as fOcol hypertrophy which progress** to onfnlir hyperplasia. and is o f*w itemoli. to tepttoou or cbolangiohapotatnaa. Tho hportrtfUc coll* crateia largo iimoiti of lighttaiaiag cytoplasm which It probably rick ia glycogaa aof aoOopteatnic rotinlao. Ia aoma cocoa. ring-shaped atroctaraa. probably representing whorl* of proliferative endoplasmic reticulum. war* ***o to tha cytoplasm of thaa* caUa. Tho hyparpiaatic oofala a occupied larger or*** mod ottaa compressed surrounding colla of tha oormol liver parenchyma. Thay ola* cootoioad th* aoma hypertrophic colli. . Tho a* laaloaa which 1 classified oa hapotomaa or ehoUaglohepatorr.** war* larger nodule* which showed evidence of ermfluone* or oama variation in coll ala*, ah*pa. atoioini or o dactular or odaooaatoaa pattara of growth. In th* obaaoc* of a**toato*la. iovoaivanaa*. aavara basophilia. mitoses or othar evidence of anoplaaio. 1 choaa to coll thooa benign tumors rather thoa moLigaon: carcisomoa. Thar* tia no avidacca of mataataala *r lovmatvoaaaa of thaa* tumors In this atudy. WATER PCB-SD0000012814 s IO-TCIT 209 I i*n naihatojUU wight iH*i wJI >i <i>m< Wltk inimu f tha Titulif rtan#aa 1> tfca cyWftaam af bapatacyt*a. tfaa I^ Bar* aaaar* bapa(atalhilir tlkraliM mAm4 m animal* af tha Hr 24-Uaalk Racrtfica parl*4a. (iU*JtZ .Gu/M* Ward R. Richtar. D. V. U.. U. S. Diplomat*. Amarlcaa Collafa of atariaary PathotofUta jp>v*w J -^'"W * 0002.1/ A WATER_PCB-SD0000012815  rw* Primary thr UiImi AncUr IMS Sac rifle* bu 5 2(0 VaraoUr Cku|i TocaI B|yrtn|hy NoMir HyprpU*U Hepttomi Dactolar HyparpUaU Ckolia|lo-HepelB> H*p*toc*lh>lar Nacroai* TX 0/11 0/11 0/11 0/M 0/11 0/M 0/M Til 4/10 0/10 0/10 0/10 1/10 0/10 0/10 rni c.J,* A 3/10 Control S/10 2/10 Vu 0/10 0/10 0/10 0/10 0/10 0/10 0/10 0/10 0/10 0/10 0/10 h -- WATER PCB-SD0000012816 * Primary Llnr LerUea Arttlw 1M Month Sacrifice ltti U!o Vintlir Cku|i Fecal Hyp* rtrophy TI 1/A 0/6 Til 2/S 0/3 Oreee Tin 5/9 5/9 V, Control 1/10 0/10 Modular HyperpUoU Hepatsma Dectolar Hyparplaala 0/6 0/6 0/6 0/S . C/9 0/5 0/9 0/S 0/9 0/10 0/10 . 0/10 Cholaafio-Hepatoma Hepatocellular Necroaia 0/6 0/6 0/S 1/S 0/9 0/9 0/10 1/10 WATER PCB-SDOOOO012817 7 212 Primary Lirar Uilou Amtei IZte 14 Maolh Sacrifice . bu Ul V*nitit CkU|< T1 1/9 TO 9/10 Owe TD *+.' i J/9 '/ focal Hypertrophy 0/9 0/10 4/9 Itodalar HyparpUaU 0/9 0/10 1/9 Vo Hopataioa 0/9 0/10 0/9 Dadiltr HyperpUeU 1/9 0/10 2/4 " Cbolaaflo-Ha pa coma 0/9 0/10 0/9 Hepatocellular Nacroala 0/9 0/10 0/4 Control 1/10 0/10 0/10 0/10 1/10 0/10 0/10 WATER PCB-SD0000012818 Vo Ll*r Uilni Arwltr UK. 213 Ttnslal hcrlflc* Uu Vtnolw Ckaaf* Toc*l Hypamophy Nodular RpparpUila tbpttaoi Ooctol&r HrparplaaU Cboluf<.H*p*tan* IbpttsuUalar Nacrotia Oww TX A$ ,' /2S Tn A,. 4/23 im ,. Cratrtl 10/27 hhl 1/23 ./'/vy' HrV3/25 10/23 *ul. 1*27 y,-^ 0/23 `A4' am 5/23 7/27 y, A, 1/.3 ,/.V 0/25 7^' 0/23 *4 5/27 'm 0/23 t/jy 4/21 'r ^rf 5/23 0/25 Ul< . 0/23 V '14/27 5/23 */'* V 2/27 V.- f-7 0/23 >J>1 4/25 %.* >, 2/23 V11 *1/ 4/27 4 < --' 1/23 0007-3^ WATER PCB-SD0000012819 " A.- >* Vuoi^. .. *^ Am? ' > OOOP.^r: WATER PCB-SD0000012820 / ji ) _. . * t .N c" . H /* RJ m tf n -- \ 6 s' r? 3: iS|S3232 0 ::x2*fi33g 353*? 4U wi :Kj 3 r555 JrUi N i :| ) i| n?\ Z lit* Vaaaalar Chaaga Focal Hicrtili in : Focal lymyhaM laUtltntloai Focal hypertrophy al hapalecytaa Nofelar hypertrophy ot hapateeytea focal glia pool Nrpe rplaala lUpiiona r i :3cCr. r i 8! r CHoUMglo-hepatenia 9CZ000 WATER PCB-SD0000012821 .* '* n ' --e \s tn * 1 V' 6 i kV ^ r P a fc S3cV ll <-- r? 2: : . 3 31 : : : U +^ : i ; : 5| 2 j i : g j 4- : 3 IM ^ s : i{ * ?i?i Vaeuola r Chang* Focal Nocroala t zz z *T * is . . Focal lymphoid laflltratloaa Focal hyportrophy at hapalocyl** - Nodular hypertrophy ol hopatocyta* c 3 Focal Blit Docl r Hy^crplaiU S* * Hopatoma Cholanglo-hopatoma 1 jk WATER PCB-SD0000012822 000738 WATER PCB-SD0000012823 Sacriflca latarval ru*i <COAtlS*4) .n r/.fc k~ TtMtlloi W UItMmI Livar UllHt la RU 218 Don Laval 4aiaaA Na. aad Sn AH hit 11J" 220** 221" 224" 233 234" 219-*l -+230" *| 203" 204" 204" 44 207" ^4 liar LwIom *8 f: a 1 II -3 If cI il 11 fi u*. i0 ' Z m' 4 -< 4 (-1 P if ii 141 44 <f. P'l n 1 2 w0e AHI 24U4 233" 2*1" 244" 270" 271" h*-**' 272"H)4-*-*+ 27$ v`H 243" H4 t? 244" 312" 2S1T 274" 27**4+v4Pt 294 297" <r*44 302' 304' 300' 311' 314' 240- iv^ftK1 r* \t ** |V-| 4 * |-l 4 4 * HP 4<tf l-l * I 4*44-4 ll L+44 U l P>1 If H)fI-' 4 - P t ) PlP' -* 4 (.! 4P-' URH c- I) oooms WATER PCB-SD0000012824 tpuf on `mmu^ j **** >1 4*T1M* >1 4-M-f **** <n nr i4U*** T lwmjmi 4 BtM( llipug 6 12 * "* _ n i>pi*p"i > 'h*o*i,*m*x m wooir __ WATER PCB-SD0000012825 Sttdu.iUud tlO-TIST lK MONTtSI *o*o NOtmitoot.iLUMOii *oo*j Ate. REPORT TO MONSANTO COMPANY TWO - YEAR CHRONIC ORALTCXICITY STUDY WITH A ROC LOR 1260 IN ALBINO R/.TS H1STOPATHOLOCICAL EVALUATION OF ADDITIONAL LIVER SECTIONS MARCH 24. 1975 I 3T NO. 641 - 06671 172 %+UUud *10 -TUT Jehtmknmi. %c Mte raoNTAM aoao MOaTNIIOOK, ILLINOIS 0*001 173 Caarga J. Ltrlaikii, Ph.D. Mounle rimiiii) 00 N. Lindbargh Bird. St. Leaii. UtimrliJlU . Dmr Dr. Lavtnakaa: Rt: DT No. Ml-04472 - Klaaepaiholofical Evaluation o< Additunol Liwr SacHnna Froa Kata at a Two - Yoar Quvuc Oral Toaidty Study ot Aroclor 1240. Wi arv aubalttiug baraartth our i arlaad Laboratory rapart dazed March 24. 1475: prepared to caawectien with tha abova rtudy. Vary truly yaura, J. C. Colondra Prandent tlO-TIST c 174 REPORT TO MONSANTO COMP ANY TWO - TEAR CHRONIC ORAL TOXICITY STUDY WITH AROCLOR 1260 IN ALBmO RATS KISTOPATHOLOGICAL EVALUATION OF ADDITIONAL LIVER SECTIONS MARCH 24, 1975 1ST NO. *41 06672 I. Introduction At tho roqnaai ul Dr. Lorinakaa of tb Monaanio Compac-', additional actlona ol '.Ivor IrotB a two roar chronic oral toxicity atudv ol Aroclor 1260 in rau II2T No. 622-072981 war# procaaaad into K 6 E atainod action and oraluatad by li|ht mlcroacopy. Tho (ollowtai report praaasta tb raaulta ol thia artady. -Aw|y. ji.w i., P ----------------------- --w !T WATER PCB-SD0000012828 jUwba/ IIG-TIIT AtniUm fm. 2. 175 . Suamarj fa seat Instances, tea spectrust a( Irastwsnt raltiod histspaihafagkal Mta|t b the lint from thla n>nltttln did eat differ significantly frees that prevtoaaly reported la ear original report dated November 12, 1971. However, there were even-benign liver tomere detected among eaves of the at the Highest traetroent Ural (100 ppm) of tba 24-Mooth Sacrifice which were wet previously reported. Tha other treatment-related Uaiosa reported are regarded as dsgsasra- tire or byperpUstlc is satore aad they are morphologic maniie ateHoar of ea adaptive reapoasa af the liver associated with biotraasformatien of the teat material. In general, the Uttar findings were coaflaad primarily to teat animal a of tha 6-, 12- and 24-Month Sa-rUicea. and they were dose-related la incidence and aevarlty. In conclusion. Aroclor 1260 appears to be slightly tumorlganlc at levels of 100 ppm whan fsd continuously In the diet tor two years. Respectfully submitted. INDUSTRIAL DIO-TEST LABORATORIES. INC. Report Prepared aad Reviewed by: D. X. Gordon. D. V.U..Pfc. D. Section Hoad, Pathology sP- * WATER PCB-SD0000012829 JUmitrn/ KIO-TIIT s I1T Ke. W4TtN Mmiimw 176 l lurt nimlB< iMIMwiil aectloaa of Unr from rote of OT Ho. 622-07291 aad tan rt-mlaitad thorn ter nUtsci mi carrlaagaa aria. V bin tabulated ay flteli|i ter Aroclor * 12*0 M tha kilnta| pagaa: I Ttart li ewideaca ol a cbimlcal offect oa tea Unr Ueb *11 eioit rrltait at Hat ol tea 12- Month and terminal (24-Moath) aaerlflcaa. Thl* eoaaliu of a hepatocellular alteration hefiaatag aa focal hypertrophy whies progreaaea to nodular hyperplaala. and in a fbw aalraala. to hanatoma* or chalaagiehapatomaa. Tha hypertrophic ealla contain lar|a amount* of light- ataiaiag cytoplaam which la probably rich la glycogen aad aadoplaamic rat;- culurr_ la ooaao caaaa. rlng-*hap*d atroctnraa. probably representing wharli of proliferative andoptaamic reticulum, wore aaan la tha cytoplaam of that* call*. Tha hyperpiaatic aodulaa occupied lar|ar area* and oftaa compra***d ur rounding ealla of tha normal liver parenchyma. They alee contained >.* am* hypertrophic call*. Thoaa tacioaa which I claaalAad aa hapatamaa or eheLangiohcpato.--.*j w*r* larger aodulaa which ahowad avidenca of confluanc* or aoma variation := call ala*, ahapa. ataming or a ductular or adeaoraaioua pattern of growth. In tha abaoac* of mataataata. lovaatvanoaa. aaaora baaophllia. mltoaaa or ot.-.ar ovldooco of anaplaala. I choaa to call thaaa benign tumor a rathar than malignant carcinoma*. Thara wa* no atridanc* of naataataala o. lavaaivonaa* of tb**a tumor* la thia atudy. tlOTlfT Ah*+tm, JU. With i (tl and 24-Hoetfa tlai ilfli .4 >177 *a b> ryt^liwi f \ ulii. let Ike U- Ward R. Mchter. D.V.U., U. S. Diplomat* American Collef* of Veterinary Pathelofiet* WATER PCB-SD0000012831 Mbut Uaur Lu1*m Aroclor UM S Uoaak Sacrifice - Kata Larioo fiSBL TI Til TUI Vtcwlir Chaafo 0/11 4/10 1/10 focal Brpotnfkr 0/11 0/10 2/10 Nodular HyparpLarla 0/11 0/10 0/10 Hepatoma 0/11 0/10 0/10 Duetular Hyperplasia 0/11 1/10 0/10 Cbolaaf10 -Hepatoma 0/11 0/10 0/10 Hepatocellular Nocrosla 0/11 0/10 0/10 Control 2/10 0/10 O'lO O'lO 0/10 0/10 0/10 WATER PCB-SD0000012832 179 4 Primary Lint Laaloaa - Araclor UU i Month Sacrl/lca till La a ion Grata TI Til Tin Vacuolar Chanf 1/6 2/S s/s Focal Hypartrophy 0/6 0/S 3/S Nodular HyparplaaLa 0/6 0/S 0/S Hapatoma 0/6 0/S 0/S Ductular Hyparplnjla 0/6 0/S 0/9 Cholaaflo Hapatoma 0/6 0/5 0/9 Hapatocallolar Nacroala 0/6 1/5 0/9 Control 1/10 0M0 9/10 0/10 0/10 0/10 1/10 7 Frlsun' Uaar Uilm - Aroclor UM 12 Uoilb SacriAta Ktti Lea loo Own T1 TH tzh VacuoUr Chaage 2/9 5/10 1/9 focal Hypertrophy 0/9 0/10 4/9 Nodal*r HyparplaaU 0/9 0/10 1/9 Hapatom* Duetolar HyparplaaU 0/9 1/9 0/10 0/10 9'9 1/9 Cholaagio - Hepatoma 0/9 0/10 0/9 Hapatocallolar Nacroal* 0/9 0/10 0/9 ISO Control 1/10 9/10 0/10 0 '10 1/U o/.o O'lO 5k0>'. WATER PCB-SD0000012834 % . 1*1 Primary Um Leals** - XmltK.I)U: . ' Termlaal Stcrlflct - tin % . Creep Vacuolar Cku| TT S/23 TB 4/23 TD ie/27 Control 1/23 Focal Hypertrophy 3/25 10/23 1227 0 *'23 Nodular Hyperplasia 0/25 9/23 7'27 1.23 Hepatoma 0/25 0/23 S/27 0/23 Ductelar Hyperplasia 4/25 5/23 14 '27 5/23 Cholangie-HapatuBa 0/25 0/23 2/27 0'23 Hepatocellular Necrosis 4/25 2/23 4/27 1/23 aioclok ism T*I)I<|Imi( WiMwl lllvcr Laalan* ta Rill 182 _* s o <5 n if 'A'O xa +- 4<*4 a**4t *u4* w-rf4 SSSS8S <1 ajSfi : :nr i: -------- r j3p*? iJj Vacoolar Chaafa Focal Nacroala Focal lymphoid Infill ration* Focal hyporlroptiy of hapatocylaa Nodular hypertrophy of hapatocylaa Focal B>l Dcl llyparplaala H |8 -5 c* ti llapaloma .'i t . 4 H/ 1' ^ Cholanf lo-hapalama tj V. Vf `* *1 WATER PCB-SD0000012837 1 !. V o o ro w U WATER PCB-SD0000012838 f> * ! f 4- C? 3J ft vxNbf rt--f FoctlHrerMll Focal tymfloU Ikflltratlaaa Focal kyyoMrofrtiy of lopt[f)i Modular Hyrtnyh of hapalocftaa Focal |ll Dari llyporplaalii ' *) ' Ibuatoma ` It c* ii Cholan|lo>lpal*ma tn fS20G 0 WATER PCB-SD0000012839 000255 WATER PCB-SD0000012840 o [f ffff 7I7f VintUr Ckaoye rMi(N>c>*iU Fil lymphoid laflltratlaoa rocal hypertrophy of hepaloeytde Nodular Importraphy of hapalocyiaa Foul pi la Duel flyparplaata llepaloma Cliolanyio -hepatoma o3 *0 w o WATER PCB-SD0000012841 AmAuM/utL tio-TlfT Jab&aAmu. Am... < rtONfUU low *0Tf *Q. tlllMOt* MMt 077 REPORT TO MONSANTO COMPANY TWO YrAR CHRONIC ORAL TOXICITY STUDY WITH AROCLOR ISM IN ALBINO RATS KSTOPATHOLOCICAL EVALUATION or ADDITIONAL UVSR SECTIONS MARCH 24. m$ IBT NO. 441 . 0t>o7: / ,.. . . . J JW4i*W lO-TlT .MttwfmM. Ah MM SaOWTSQO MM NO(TMI*OM. ikUNOS HIM July 1. im 078 Gaoryo J. Lartaalcas. Fh.D. 100 N. Uadbarfh Bhrd. St. Louis, Missouri 43144 Ra: IET No. 041-0047} Hlalopathnloflcal laaluatloa ad Addmnrsl Livar Sections from Rota ol a Two - Yaor Ononis Oral Tosldiy Studr erf Awthr U40. Wa ara aiitiwlttlng harawllh oar i reload laboratory rapert dMad March 24. 1079, pcsparad is ronnorttan with da abora study. Vary truly yours, J. C. Calandra --s. JCC/fd WATER PCB-SD0000012843 ;UihJ tlOtIST ` ' *- - ` 7f REPORT TO MONSANTO COMPANY TWO YEA* CHRONIC ORAL TOXXITY STUDY WITH AIOCLOI 1260 IN AUmO HATS HtSTOPATHOLOGICAL EVALUATION OF ADDITIONAL LIVER SECTIONS MARCH 24. I'm IBT NO. Ml . 06472 L Introduction At the roquost of Dr. Lorinakaa of tho Monsanto Company, additional sections of '.Ivor (rota a two - r*tr chronic oral toxicity atndy of Aroclor 1260 in rata (IBT No. 622-07200) wore processed into H 6 E ataiaod ioctiona and evaluated by light microscopy. Tho following report prolonta tho roault* of this study. .* ' ty -`ynp"o*-W* ' bin 000253 + MMOMNOadC B. WATER PCB-SD0000012844 lldthf tlO-TIST ' -'Vi> 2 fladta** tm faa Nrar *om da rrtntellM dU aat dttar iHiflOMHly few that prnrtwty naat hi mm aaifftawl rapart hM H--liir U. 1171. Thara an nm hpattau hheh4 tmaaf aaraa ad tha aitaili u h hlgbast triafaal Vrval (1M ppaj at tea H-Mwtti Sacrlfica. No Impaw cellular cirdaiati am abaaired. Tha athar traafaat related laaiooa reported ara regarded u defaaarattre or hyparplaatta la natwa aad they ara arphologtc aalfaatadaaa ad aa adaptin raapeaae ad tha bvar aaaodatad arith btotranafarmattoa od tha laat MsMrial. la general, tha latMr fladtage were rnnflaart prtaarlly to Mat --`~t*t ad tha h-, 12- aad 24-Uoeth Sacrifice#, aad thay were doaaralatad la tadrtaaca aad ea rortty la eoaelaaiea. AracVar 12M daaa not appaar M be carcinogenic a rata fad far two yaara at Wtrala ^ M aad Including 100 ppa. Raopoctfully auhadHad. IMDUSTUAL BIO-T1ST LABORATORIES. INC. Report Prepared aad Raeiaoad byt 0.D.E. Gordoa. D.v. U. . Pb. Sactloa Haad. Pathology i WATER PCB-SD0000012845 WATER_PCB-SDOOOO012846 U'AjR.euMk Vtx4 R. Klehaar. D.V.U., \L 5. DtltMMM. Amrteu CU*f o( VMtrtitry PiriMtofiaw rg^^1 * -4r>v *~ /T V.5, * 000262 WATER PCB-SD0000012847  , . ' V \.k;- \ v : /r- * aiy IMt Uit - AimIm mi-iff' i-* ';v. 1 Uattlaariflc* - Kata - ' Uelf Vacaelar Oaaf* Foetl KoMtr Hfptrpltil* H paten* OwttUr HyperpUaia Chelaafia-Haparoma KapatacaUalar MacraaU TX /II 0/11 0/11 0/11 0/11 0/11 0/11 Til 4/10 0/10 0/10 0/10 i/io o/to 0/10 Tni 1/10 2/10 o/to 0/10 0/10 0/10 0/10 Cental 2/10 0/10 . 0/10 0/10 0/10 0/10 0/10 t WATER PCB-SD0000012848 "tjt' V. `* ?'** ., * .*** J*1V- '* , '.\4^ *; "VV* <- -1 - V ' *- .. 4 * - 114 \a . . /- ->. Pwtearr L4m* t t<-- Anclat wiis>4. 4 UmA SmtUU* Iw Ltw ' fin VtCMltl Cktlft T1 Til tux 1/4 i/S s/s rosi Kry*nfhf 0/4 0/S 3/S N4*Ua HrporpUaU 0/4 0/S 0/S 0/4 0/S 0/S Strain Hr?rpUla 0/4 0/S 0/s Cb*lu|ia-Utplan> 0/4 0/S 0/S lb|tMtlliUr Ntcmlj 0/4 1/S 0/S Control 1/10 0/10 0/10 0/10 . 0/10 0/10 1/10 WATER PCB-SD0000012849 CHb \. *rimarr lira* Utslaas - Aroclor UM i Meath Sac rifle* - Rats L*lcm Own TI T3 TSI Ccctrol Vacoola* Chaag* i/9 5/10 5/9 1/10 Focal Hypertrophy 9/9 0/10 4/9 0-1? Nodular HyparpUsiHapataoaa 0/9 0/9 0 '10 0/10 l, 0 '9 3 10 o :o Doctalar Ryparpiaala Cholaaflo -Hepatoma HrpatocaLlular Nacroala 1/9 0/9 0/9 0/10 0/19 0/10 i/9 0 '9 0/9 ! 10 0 10 o :o WATER PCB-SD0000012850 096 . Primary ZJtra* Laaiaaa Araclar UM Tarmiaal tacrtdaa Kata ioe " Prana TX TU im Vacuolar CAaafa Focai Hrpartrayky No4iUr Hyparplaoia tns iUS o/zs *.'23 10/25 -5/23 10/27 11/27 7'27 Hepatoma 0/25 0/25 5'27 Doc tular HyperpLaaia 4/2* 5/25 14 '27 Cholaatla- Ha pa iotu 0/25 0/25 2/27 Kopatacelhilar Nacro * 4/25 2/25 4 '27 Control 1/25 0/25 1/25 0/25 5.25 o . 1 -2 WATER PCB-SD0000012851 *n t * 11 IS 7S n?{ VttMUr CImh racilNiciHti Focal lymphoid lollliraUaaa Focal hypertrophy ol bapalocylaa Nodolar hyprffoffc> I hapalocytaa Focal nll Duel Hyporpiaala llapalotna Cholaapla hepatoma l M l **|an ia net nK |naon po>r/|Fp l f eenw |nT *l I 80 WATER PCB-SD0000012852 L. ) ii 1 . f" SUV>S -* -iS|Si(SS"S lilif hh?i r i i n?l J r Vtmhr Cku|a Focal N>criti Focal lynipboM lallltraUooa > 3 7 *t Focal hypertrophy of bapatocytaa Nodular hyparf#afl>y f bopatocyta* Focal Bllo duel HyporpUaU, \ n F ... 1 Ilopaipnu. . ' .! ChoUaylo-lioygitttna | ; \f 1 ? . . i r Jv & WATER PCB-SD0000012853 J WATER PCB-SDOOOO012854 WATER_PCB-SDOOOO012855 AXOCZ.CB 1240 TtbuUtiM ot MWidl Unr Loilm la Rata 091 SacrUlca Interval Ooa* liii L*rol i talma No. and St* i $ 4a >iU -- 2i 0V*b 11 C-- 1 *4 4 u= % . V0 i> zi It ii A0j Livar Lailau >4 It a* r * 2! H H || VVI i 5 1 9~ i i i ?*j 3w 1 i l 1 i,i riaal Icooaowdl xn '1 1 i Am I1 , ` , 2127 213" * 220" 221" 224" 233" 234" 219" ** - 230" - 203" 204" 20a" 207" * * 24IM 233" 241" 244" 270" i 271" 272" 273" 243" * 244" 312" 2317 274" a 273" * 294" i im ** 302" I 304" 1 300" l 1 311" 1 314" 1 240" t 1 l p * p ** - i i ** p* *** ** j , \ p p * * i' p* ii ** A t p p p - 1 p1 * i p ; 1* - p i 1i " - WATER_PCB-SD0000012856 ooo ti t* 3 I a a a t4A * 9 IfS ** A-pA tJfNtf 3A SA aO rrrrrMrG* mm *4 w Ut z zi s* rtf 3: Jfs ?! m a. | Victwbr CUn|a I'octlNuiroali *0 *0 Koctl lyrn|.hulJ lalllitaihma Focal hypertrophy ol hvpitocyta i Nodular hyportrvplar ol hapolocyla* Focal |ll Duel Hyporplarta ifc i*\ Si a ri I fop*loan* Cltol* agio-bepalonia rsi I l WATER PCB-SD0000012857 Lotion VicuoUr Change Focal Hypertrophy Nodular Hyperplaala Hepatoma [>ictuiar Hyperplatla C ho Langio-Hepatoma ` Hepatocellular Nocrotia Primary Lir Utloiu - Araclor lUt Tinniul licrUle* - Item M if3 m I2 Oroap 1 Till 12 3 1/25 2/25 5/25 1/21 2/20 A/23 8/27 7/24 10/27 0/25 0/25 3/25 0/25 0/25 0/25 0/25 1/21 4/20 10/23 A/23 4/20 9/21 0/20 0/23 9/27 6/24 11/27 2/27 3/24 7/2A 0/27 4/24 5/27 1/25 1/25 A/25 2/20 5/21 1/27 1/24 14/27 0/25 0/25 0/20 0/23 0/27 0/24 2/27 1/25 4/25 0/23 0/20 2/23 2/27 11/24 A/27 093 Control 12 1/21 0/24 0/23 0/24 1/23 0/24 0/23 A/24 5/21 0/24 0/21 0/24 1/23 1 : First evtluotiai of original oUdoo 2 - Re-evaluation at ori|in*l tUdee 3 - Eviluaiion of additional huso oocttoao WATER PCB-SD0000012858 MM IO-TIST VrlMiT U*ar taaitmm - AtOCUM - 1240 Tarateal lacriflca - -- -TlllUtll * Oli^lMl SJdM Vacuolar Qaaaga focal liypartraphy Nodular Hypaaplaeta BUa Duct Kyparplaaia Oiolanpo-hipilnBi Hopitona A-I 2/25 /2S 0/25 1/25 1/23 0/25 A-2 2/20 4/20 4/20 2/20 0/20 0/20 A-J 7/24 4/24 5/24 2/24 0/24 4/24 0/24 0/24 4/24 4/24 0/24 0/24 WATER PCB-SD0000012859 WATER PCB-SD0000012860 IO.TIST i 6 WATER PCB-SD0000012861 [ c C\ t ! \ I f \i r ;'v WATER_PCB-SDOOOO012862 E X H I B I T WATER_PCB-SDOOOO012863 . ( ^6 tio-TisrOrAuUaL fnUvttmm. She. i*t# rioMuai *e*e NOITHIHOOI. lkUMO tMt] 239 REPORT TO MONSANTO COMPANY TWO - TEAS CHRONIC ORAL TOZXCITT STUDY WITH AROCLOR IN ALBINO RATS KBTOPATHOLOGICAL EVALUATION OF.ADDITIONAL LIVER SECTIONS MARCH 24, 197S I3T NO. 441 - 04*72 WATER PCB-SD0000012864 210 t SmiatBiit tIO TUT ftrifirtTfrif She. 1*1* PaONTtOI *OAO NOirxiieo.iiuMii tNti March 24. im George J. Ltriuku, Ph. D. MouMsto Company BOO N. Lindbergh Bird. St. Loili, Missouri 63164 . Dear Or. Lerinakaa: ' Re: IBT No. 641*06672 - KLstopathologleal Ermluatioa o< Additional Lirsr Sections From Rata ol a Two * Tsar Chros&ic Oral 7oatc.tr Study ot Aroclor-6a6. I7.S1- We rs submitting herewith oar Itboratorr report dated March -1 1975: prepared In connection with the shore study. Very tralr yoars. J. C. Calaedra President WATER PCB-SD0000012865 Ari >* ,< IIO-TUT Afrssfcini. .W i *41 REPORT TO MONSANTO COMPANY TWO - TEAR CHRONIC ORAL TOXICITT 3TVDT WITH AROCLOR 1264 ' IN ALBINO RATS histopathological evalcaticn or AOOITIONAL OVER SECTIONS MARCH 24. 197J 1ST NO. 641 06672 I. Introduction At tha roquaat of Or. Unailai of tfci Monauto Company, additional sections of Uvar from, a two year chronic oral tonicity study of A roc lor K66 '.ly T :n rata lIBT No. 621 -07276) ware procaajad into H 6 E stainod aactiona and avaluatad by light microscopy. Tha following report praaanta the raauita of this study. WATER PCB-SD0000012866 lO'TIST 2 242 --( In mot truaanl-Nl>M2 Mrtcyi>ih|lnl fladii|i la the liver from tide re-evalnntioB Aid aet differ aignlfUantly from that previously reported la oar ori|iMl report dated November 12, 1971. Hiieint, there were three lenip Ueer tamore detected amopg three of the ulsali at the highest treatment level (100 ppm) of the 24-Month Sacrifice which were aet previously reported. The other treaeaent-reletad leeioaa reported are regarded aa de generative or hyperplastic la aatore and they are morphologic manifestation* of aa adaptive reapoaae of the liver aaeociated with biotraaaformatioa ai the teat material. In general, the latter (lading* were confined primarily to teat animal* of the final eacrlflce and they were doee-related la Incidence and aeventy. In conclualoa, Aroc lor appoars to ho (tightly tumorigeaic at level* of 100 ppen when fed contlaouely in the diet for two year*. Respectfully auhmltted, INDUSTRIAL BIO-TSST LABORATORIES. INC. Report Prepared end Reviewed by: O.C. Cordon. O.T.M. .Ph.3. Section Hoad. Pathology WATER PCB-SD0000012867 IO.TIST t IBT No. HI-07Z*I Uoaiuto j l kit* uunlitd idditlAMl McdMi of Utn tM nil of Staff Number 022-07240 ud hare t*-*nhut*d tbam (or eeidaace of tabulatad my flidh|i far Aroclor f&t M tha following pages. ; inc Thors la oridooeo of a chemical affect aa tha liver which l* moat pronounced at ttma of tha terminal (24-Month) sacrifice. This conaiita o: a hepatocallular alteration beginning aa focal hypertrophy which progrv to nodular hyperplasia, and la a few animals to hepatoma or cholani'.ocepatocna. Tha hypertrophic eella coataln lerfe volumes of Ught staining cytoplaem whteh i* probably rich la glycogan and eadoplaemic reticuhan. la soma sections, .-.c,-- haped truetore e, probably representing wbarls of proliferative endoplasm..: reticulum, ware taaa la tha cytoplasm of thaaa cello. The hyperplactic nodulee occupied larger areas aad often compressed eurrounding cells of the normal Urer parenchyma. Theee nodules also contained the same hypertrophic cell*. Thooo losioas which 1 claesUled as hepatomas or cholaagiobepatomas larger aoduloa which showed oridoace of coafluence or lomt variation :n cel! i.i;. (hops, staining or a ductular or adenomatous patters of growth, hi the absence mataetaels, laraalvoaoae, severe basophilia, ttutoaes, or ether evidence of ana plasia. 1 ehosa to call thaoa beaiga tumors rather thaa mallgnana tumors cere.n- onail. Thors erou so evidence of metastasis or larailveeeae of these tumors in this study. WATER PCB-SD0000012868 ttO-TIST 244 ' U aha*W- ba |itortWH mmi ftlMlafiair- lamanim ih >>iii nf nitiir trlft**4* v--i -nn ^ jih1*1* With axcaptioa at tha raneltt du*|ti la tha qrtafUm of hapatacyta. tha r\ mora tanra hapatacallular aUaratiaaa wara coaflnad to aaiaaala al tha 24- Uattk SacrlAca. Ward R. Rlchtar. D.V.M..M. S. Dipl., Am. Collaga Vat. Path. i WATER PCB-SD0000012869 wv Prlsurr Unr Lnou - Arocler 126* X 1 Month Sacrifice Rati Utlott Gr2 TI Tn TOT ' .cuclar Chany* 0/8 2/10 1/9 Focal Hypertrophy 0/8 0/10 0/9 N'ouolar HyperpUaia 0/8 0/10 0/9 ` Hepatoma 0/8 0/10 0 '9 Ductuiar Hyparptaaia 0/8 0/10 1/9 Jholanyio- Hepatoma 0 '8 0/10 0/9 Hepatocellular Necrosie 1/3 0/10 0/9 5 2 4l> Control i. 10 c 'i: o i; ,9 * * : :u :n 000284 WATER PCB-SD0000012870 m yr Primary Liver Laalona - Aroclor 126# - ft 8 Month Sacrlflco - Rata La (ion Group T1 th TIH Vacuolar Chang* Focal Hypertrophy 0/10 0/10 0/1 0/8 0/9 0/9 Nodular Hyparplaala 0/10 0/8 0/9 Hepatoma 0/10 Uucralar Hyparplaaia 0/10 Cho Lang io-Hopatorca 0/10 Hepatocellular Necroeia l .'10 0/8 0/8 0/8 0/8 0/9 0/9 0/9 0. t Control 1/10 0/10 0/10 . e/io 9/10 0/10 1/10 000285 WATER PCB-SDOOOO012871 7 Primary LWar Laaloaa Aroclor 12M l 14 Month Sacrifice - Kata Lealoa Vacuolar Change Focal Hypertrophy Modular Hyparplaaia TX 2/10 0/10 0/10 TU 4/10 0/10 0/10 Group tm 0/7 % 1/9 *1 0/7 Hapatotaa Duetalar Hyparplaaia Cholangic-Hapatotaa 0/10 0/10 0/10 0/10 1/10 0/10 5/7 .0/7 0/7 Hepatocellular Nacroaia 0/10 0/10 0/10 247 Control 1/10 0/10 0/10 0/10 . 1/10 0/10 0'10 pV T 0. < '<- WATER PCB-SD0000012872 Primary Lirer Liiiou - Aroclor 12M Terminal Sacrifice - Rata g * 248 Letton Vacuolar Chao** 3% p* m UTa, 3 1t Group TQ *> /30 Tin 3/20 t1 Om f lControl 'Ya:^ 1/21 '- Focal Hypertrophy Nodular Hyperplaela Hepatoma Duetular Hyperplaela C ho laa* to - K* patoma Hepatocellular Necroaia 2/11 1/30 1/20 0/23 ,, 0/JI 2/30 S/23 1/23 0/ll*//^7 C/30 2/20 0// <' <U 0 23 i/n^Sr/Tn 3/10 3/20 Yta 1 5/23 o/ii 9i* 0/30 */)* v.*i 1/20 %*, 3 23 >/3> Yh'Ui 1/10 G> 0/20 I* I*, 1 23 <3 3 X ' . `. , / 0v .H I4* i<*( a/^ -- u a " y ^ ^ ! -* 4 V 'I 3^ 4 A O'ai ' -- 0007.87 WATER PCB-SD0000012873 fr -f* nn og 4 9_ o ^ *4 e 4w y* # w <4 w eM4-4-4-<a4 < e s < i s : : :N 3 : : ! . on Ibbb V i i: > X 2 4 . u*A O O o N> CD CD V _JI o rs? J3 i> ***4 9ft> *4 *4 O*4' ->| OO'OOMff' < = = ->: 0'0'9'0'9>Wugi Q'ft*4ft<ftftft a a -. i S ? s E teyC:n,c vacuole . htMtocmi. Focal Mcrsili ol hapatocytee Focal lymphoid Inflitralluna Fecal hypertrophy of hapalocylaa Nodular hyparplaala of hopalocytaa Fecal bile duct hyparplaala & h -p Eg 37 r* '-it = t; Hepatoma C.holany lo-ha paloma lioliculum Call Sarcoma] malaatallc Mammary tumor, molaalallt: Tulanitut I.IHIJ, local N <0 WATER_PCB-SD0000012874 AROCLOR . 12**.'" Tabulation erf Iadtrirfual Unt Laalaaa la Rata !0 250 SaerUica Doaa Animal) Intarral Lara l No. Lira Uilaai and tkSax in ibi ?$ 2* *8*2! -- 1S 4= K -2 Ui9<.* f n M i k50 * *1 &! a 9 - 1z 8 4ll 11 4&9 ? 9 i j SJ i -1-l 3 Si U'tl i_li ; i Month I C-I tailM ! <PP*a 1012" ; 1011". 1014"' ,i p i x'.'.'J, 1015" 1 1026F, 1027" , 1028" ] 1029" ; 1030" l , c-n io41m: ' lOppm 1042" 1041-; 1048" I0S*F ! 1057" I 1058" , 1039" c-m io7im . lOOopm 1072" 1 1073" : 101J" I ' ^ *^lS I0S6F I >i- i?.J. Y 1017" ' 1088" 1089" 1090" Control 801M 892" <>S 803" 804" 803" 816F 917" 818" 819" 820" 000289 ft *? . WATER_PCB-SD0000012875 AROCLGR . Uift~ rtbttUtiu of Ladivldatl Uni Ualoaa la Kata U T o U n g e c U iU , focl '# - . - T>*v*'-n>v 000230 WATER_PCB-SD0000012876 AROCLOR . I24a; TihritHw af laRlvUaal Uftr Laalaoa la Rata 252 12 1. WATER PCB-SD0000012877 i-- TT-T t J to n 11 -f Is -----------U ---------------f-------- f----- 044iHHOoodjiiiiii^uiWiIiI,Kfruiui rr ;< ; ;^ i ; i * tt 1 JL TT 1 jCytopUimic vicnoliiioa of iMpttocytoi Cucil oecroela of hopalocylaa "T" T" Focal lymphoid laflllralloo Focal hypertrophy of liapolocy los 1S I* TT T t _1_ Nodular hyparplaala of hopatocytaa Tocol Mia duet hyporploalo I'apatoma E 1` *Z V w Cholan|lo-hepatomo Reticulum Call tlarcoma| metaalatie Mamniiry tumor, iiivlualalic I - I i it* I i * i*i , I* >i I u< Co WATER PCB-SD0000012878 I AHOC LOS TabaWttaa sf Mlfltal Unt UiUu la Bats {$4 WATER PCB-SD0000012879 .-> ___ 1 S acrifice (V s* Jy f t ^VjT rf l rv *i* s *n 3 s i * -S 1 Cylc^Uwnlc Vkcuolaliaa of hapatocylea Focal aacvoala of hapalocylaa Focal lymphoid laflltratlraa Focal hypertrophy of hapalocylaa E2 Nodular hyperplasia *0 of hapalocylaa J Focal hits duct hypo rplasla r o ft lla pa laina Choleagio - hepatoma Mottculum Call Sarcoma molaaUlIc Mammary tumor, mUuilc T Idiiitei ld^iit, fui.it M cn CA w AROCLOO - lih fc - i iM iT Hm at la d vid u a 1 L iv e r La alone la Rata f WATER PCB-SD0000012880 DnAuMtoal IO*TIST 2abo*ito**L. 3*0. 110 MONTMI lOAO . NORTH MOO. itUNOl* MMt 155 REPORT TO MONSANTO COMPANT TWO TEAR CHRONIC ORAL TOXICITY STUDY WITH A ROC LOR 1242 IN ALSENO RATS H3TOPATHOLOGICAL EVALUATION or ADDmONAL LIVER SECTIONS MARCH 34. 1975 IBT NO. 641 - 06673 / V*; ,J^W^r'w-9 nr 00029;' WATER PCB-SD0000012881 jWfrrf/W 10'TlfT ftrifirwifrlMM %c 1IM PftOWTAOl tOAO ' NOirMitoox. nuNOit #ooit 156 Otarf* J. Lerlaakae, Ph.D. Monaaato Coapeny tOO N. Uadberyh Sled. St. Louis, Missouri i)lU Door Dr. Lorlinfcn; Roi IBT Mo. M1*0M72 - Histopsthological tnlwtia at Addlttooil Liver Sections Froa Roto of a Two - Toor Chrome Oral Toxicity Study at Aroclor UU. Wo or* rubaittlng herewith our revieed Uberetory irport deud March 24, 1475; prepared la connection with the above rtudy. Very truly yours. J. C. Calaadra President WATER PCB-SD0000012882 Wn/ BIO-TIST ' * JU 157 REPORT MONSANTO COM. ' TWO - YEAR CHRONIC ORAL TOXICITY stcoy wrrH aroclor imz D? ALBINO R>TS HISTOPATHOLCOICAL EVALUATION OF ADDITIONAL LIVER SECTIONS MARCH 24, 197! IBT NO. 641 - 06672 L Introduction At tfaa r<qu<t ai Dr. Laa-.nakaa ol tba Moaaaato Company. additional aacr.oaa of lirtr from a two - mr chronic oral toaiclry attady al A roc lor 1242 tn rata (IBT No. 622-072981 worn proeoaaad late HI I atatsod wetitai aad avaluatad hr light tnicroacopy. Tha following rnport praaaata the raaulta of ihia atudv. WATER PCB-SD0000012883 JUiW BIO-TIST 158 IL Summary la iot uutucu, Ik* apactrun of troataaat-raUtad lil*b|i*t1>aliiglcil ta th* lirar from this r*-evaluation did not differ significantly from tha: pravioualv raportad la oar original rapori datad November 12. 1971. Koatm, there war* three benign liver tamer* detactad thro* of tba animals at th* highest traatmaat laral (100 ppm) of th* 2 4-Month Sacrifice which war* sot previously raportad. Th* other treatment-related la Slone raportad ar* rogardao as da- goaarativo or hyperplastic in naturo and they ar* morphologic manifastations of an adaptive raaponao of th* liver associated v.th biotranaformarion of da* tas: matariaL In general, tha latter finding* war* <*onf:aed primarily to t*st mtma.s of tha final sacrifice and they were dose-r*lat*d la Inc id*nca and sasentv. In conclusion, Aroclor 1242 appaars to be slightly cumortgenic at level* of 100 ppm whan fad continoualy in th* diet for two years. Respectfully submitted, INDUSTRIAL. BIC-TSST I-ABCRA7QR15, NC. Report Prepared and 3viewed by. D. E. Cordon. D. V.M. .Ph.3. faction Hoad. Pathology Report Approved by; Manager. Tjncolog'J if WATER PCB-SD0000012884 < 1 Wmlkml 110-TIST IBT No. 622 >07296 Monsanto im. } ' f_ 159 t havo examined eddltiocnl Mctlou of liver from rat* ti 9tdy Number 622*07298 aad have rs-evaluated them (or evidence of carclaoyoeoals. I hato tabulated my (ladlaii (or A roc lor 1242 oa the following pa|M. There it erideace of a chemical effect oa die liver which la moat pronounced at time of the terminal *24-Month) sacrifice. This coaaista of a hepatocellular alteration beginning as focal hypertrophy which progresses to nodular hyperplasia, and la a few aaimals to hepatoma or cholaagioheaatesta. The hypertrophic cells contain large volumes of light staining cytoplasm which .i probably rich in glycogen sad endoplasmic reticulum, la some sections. .-mgthaped structures, probably representing whorls of proliferative endoplasmic reticulum, wore seen in the cytoplasm of thoso colls. The hyperplastic nodules occupied larger areas sad often compressed surrounding cells of the normal liver parenchyma. These nodules s'.so contained the same hypertrophic eolla. Those lesions which l classified as hepatomas or cholaagtobepatsmas wore Isrger nodules which showed evidence of confluence or soma variation :a cell stao, shape, staining or a due tula r or adenomatous pattern of growth, la the absence of metastasis, invastveness, ee-.-ere basophilia, mitoses, er other evidence of aaapiasis, 1 chose to cell these benign rumors rather than malignant turners carcis;rii . There was no eviieoce of metastasis or tnvaelvsnass of these nmors :.-.:s sndy. J* - ail L WATER PCB-SD0000012885 Wi.fciW IIO-TIST JU ' 4 i 160 With aacaptiaa at tha vacular ckafM in tha rytnplht tha ora aavara hapahirallnlar ahanttawa wara caafiaad a aataala at tha 24-Uaalh Saciiflca. Ward R. Rlehtar. D.V. U..U.S. Dipl., Am. Collaga Vac. Path. f - . ' r'" .. ' i-V^Saifcsate*-. ________ _____ Of)Qr: WATER PCB-SD0000012886 Prlnirr Lirar Laaiooa Aroclor 1242 S Hath Sacrifle* kata- La (ion Crou TI Tn T1S Vacualar Chaag* 0/S 2/10 1/0 Focal Hypertrophy 0/8 0/10 0/0 Modular Hyparplaala 0/S 0/10 0/0 Hapatoma 0/S 0/10 o/o Doetular HyparpLaaia 0/8 0/10 1/0 Cholxnf Lo> Hapatoma 0/8 0/10 0/0 Hapatocallolar Nacroal* 1/8 0/10 0/9 1S1 Control 2/10 0/10 0/10 0 10 C-'10 O-'IO O'lO 0^0301 # WATER PCB-SD0000012887 Primary Unr Laiioas Aroelor 1242 4 Month Sacriilca - Rats . La slow Oroug t: tu Tin Vacuolar Change Total Hypertrophy 0/10 0/10 0/1 0/8 0/8 0/9 Nodular HyparpLasia 0/10 Hapatoma 0/10 Duetular Hyparplaaia 0/10 Cholaag io-Ha patotna 0/10 Hapatocallular Naeroais I MO 0/8 0/8 0/8 0/8 0/8 0/9 0/9 0'9 Z *? 0/Q t Control 1/10 0. 10 ' 0/10 - 0 10 0. 10 0M0 I 10 0003cr 4 WATER PCB-SD0000012888 Primary Liver Utiou - Aroclor 1242 12 Month Sacrifice - Rata Lation TI Vacuolar Chang* 2/10 Focal Hypertrophy 0/10 Nodular Hyporplaaia 0/10 Hepatoma 0/10 Ouctular Kyperplaiia 0/10 Cholanglo - Hepatoma 0/10 Hepatocellular Necroaia e/io Tn 4/10 0/10 0/10 0/10 1/10 0/10 0/10 Croon tm 0/7 1/9 0/4 0/9 0/4 0/9 0/13 7 163 Control 1 / to 0/10 0/10 0. 10 1/10 0/10 3'13 000.^03 WATER PCB-SD0000012889 Primary Liver Leeloaa A roc lor 1242 Terminal Sacrifice Rate J64 Legion Vacuolar Change Toe el Hypertrophy Nodular Hyperplaeia Hepatoma Due rular Hyperplaeia Cholanglo- Hepatoma Hepatocellular Net oeli TI 7/11 2/11 0/11 0/11 1/11 0/11 1/11 Greg TU Tin 3/10 9/:o 1/10 /:o 2/10 3/20 0/10 2/:o 1/10 i/:o 0/10 ' i/:i l 10 o,;o Control ! :3 i :j l :i i :: 5 :i o :i WATER PCB-SD0000012890 796F , 797 i 799 " . *00" i . Coatrol J6M * 37" ja -. 76F 1 77" * WATER PCB-SD0000012891 0* 0 2. 1 if on p O rn hn -- i2 ii -I = , a OOOOOOOOO S5 Q5 Bosifl-la:Qo*I1Ou?li->wI|^r.s*: j.| OOOOOOOO IPIA4AIM+*** OOOOO' W U N 14 N 4mUlW|4M O-fi P >J O' .;_'J1: J_:._ 1 oOoo iClN M 1 a =x VC g>. >i & o | Cytoplaainlc vacuola- 1*4* P( Focal oocroala of hopatacytaa H0 1 I + Focal lympbold Infl Itratlona e. l> Focal hyporlropky of T 53 bapalocylos Nodular hyporplaala of bapalocytaa cM rs fc r i cS Foul blU duel by^rplatlt s S' lUpa|om* Cliulanglo-hopaloma luilculum Coll Sarconu motaalallc Mammary tumor, malaalallc ... . 1 lulaii||oclaa|a. focal WATER_PCB-SDOOOO012892 >ald ita lic AROCLOR . 1242 Tabulation of Individual Uxr La.Ion* U Rata WATER_PCB-SD0000012893 M Sacriiica tnie rvml AHOCLOA . 1342 Tabulation of ladlvidnal Lirar Laaloas la Kata 168 >2 Dot* lAnima Laval No. and Sax u ; l 11 i U o a8a. n. e= *'3 iao lo l Ll 1 Lirar Losiaaa o 2L tauo r u ? il i n; ii i ir u 24 Month jC-I ; 1PV" 564M 546" 591" 567" 56Q. 572" j 576" i 580" .j 55a8o8." j 3 95" | 601F | 604" j 605" 582" | 583" | 607" I 608" I 609" i 613" 621" 627" 628" i 631" 634" 635" 606" 610" 620" 626" 633" *I II 1 Si i'8 .i WATER PCB-SD0000012894 i ' DT7VX3T s AROCLOR U 4I T a b u la tio n of In d iv id u a l L ir a r Laalona in Rata 000303 * 0 1 Q0 i rJ?: 0OICif>-00004l<0'Pftftafe'4Ui^^fW^(9>i<UiUi (MmwNNhM-4WO-4RO(R-A'I9'Um40`U>^WmOi4N-4U< i :i) ::: : :::: : :: :K sp e ? >| h o. r 3 * fr * of hapalocylas Focal aacroai* of hapa tocytaa foctl lymphoid Infiltration* t* Focal hypertrophy of liapalocylaa o i Nodular kyptrplaelt n E1of lupatocylai * 1 t Focal bila duel hypo rpla la r e lla pa Ioann .. a - Cliolanclo-hapaloma ' lUllculum Call Strcomi mol* (italic at 40 Mdinniry tumor, IIMlAllatlC *1 1 1.4 *i l.l . 1 *1 WATER PCB-SD0000012895 AllOC LOR 1242 TibuUttog of bflTidut Liror Ltiltu in Rato 170 Soertflct C4 Month Doo UaimAl 1 Laral 1 i No. j a4 i!5Sos S. a H s* s oa "$ si , U ii E -- i l jc-m *5 00. *0 0u0 ia g 740M 0V0J (a jlOTpptn 722" 724" 744" j 746" l 748" 768T 771" 77S" Ext" 4-6-f i 733" 765" 1 7G6" 44 iJ 767" 776" i 773" i 786" 4** : 789" 793" ^644 i Ext" 44 ^Control i : i 4M II" | 12" I 21 23 33 i1 J345" I ;*-. - 4*r i 447,.". 51" i k-! **"j ! 59" ! 62" | . Llr<r LmIooq a 0 1* Is 0 *> ti u S a H |i Si |i ia r090 (a 2 V00 h- 1 a ** P 4 P P * P 44 4P 4P 4P . * 4 *P * p p i i i o ?o C 1 : i i * . 1 ! ! : Ii - ou <5 2 Ua II sa =; u ' WATER PCB-SD0000012896 24 M onth C o n tro l 6JF **+ M odarata la aavarlty --+* M a rk a d la a a r a r ify P Proaoat. ao grade 3P AE i: SEH |Cyloplamlc v*coUiion of hapilocytti Focal aecroata ol hapalocyiaa Focal lymphoid lallltratlooa Focal kypanrcphy ol kepelooylaa Nodular hyparplaaia of Ibepalocyloa Focal bile duel hyparplaaia s. I> 7P H ~o ra u llopatoana ClioUngiO'bopaloiiia Italltuluni Call Sarcum* malaalallc Mammary tumor, IliaU alalic 'I * l.tl((M-1 l.i j i.i , foi .11 i & i- WATER PCB-SD0000012897  \.- SnlaCfal JtUmftm*. Jkt ttM npNTMl MW ho*tmi*o. uxino> --11 REPORT TO MONSANTO COMPANY TWO TEAR CHRONIC ORAL TOXIC:TT mrDT with AROCLOR tMI IN ALROIO RATS HUTPATHOLOGICAL EVaLCATI ON or additional lt'er sections MARCH 24. 1479 1ST NO. 441 - 0*472 098 /' WATER PCB-SD0000012898 MAurvtl io-tist * Akl 099 NM nONTMl toao % Nomueei. iujMO<a mm Jul/ 1. WTJ C|| J. t uliirtH fk.D. MO N. Uad3gfcBlvd. S. Louis. Mil--rl 4JU4 Dmt Dr. UMukui ' l OTIto. *41-44472 HlsMpadialotlcil Ivaluatiao at AddMaaal Ukam lcta* Fraa >t at a Two - Yaar Qmlc Oral Toalctty Stud-, at Arochw 1242. . Wo aro inbolBlaj hotooKi our ravioad laharaiary report dated March 24, Ifn. prepared la coaoscaea with dw above atody. Vary My your a. ^ X. Td jTc. rllaadra < / JCC/U WATER PCB-SD0000012899 10-TIST f 100 UPCIT TO MCKSAJrrO COMPACT TWO - TXAS aatONIC OPAL TOanCITT rrtmr vm aaoclos uc W AU0O RATS KBTOPATHOLOGtCAL CTAUJATION or additional uvn srcnows MAKCH 24, HT 1ST WO. 441 04*72 L latrodactloa At dlt ttfMit ot Sr. Urtukti ot Ik CtBftir, tddlBoMl Hctlau l Unt (tub t tv* r**r tkroalt oral toxicity otady o' A roc lor 1242 la rati (1ST Wo. 422*072 90) war* procooood lata Hit itainoC toctioc* . i ovotoottd by kfht mlcroocopy. Tb* following report protoat* tfeo rooul.a t* thi ready. WATER PCB-SD0000012900 J' IO-TI9T * flM bv that prvrlMMlf 1ml UM a *a Tba 191 .:v a*itm. t**t aMtl. h paail, A* <d tba final Merita a 1242 to aa la ba ppa. DBOSZSIAL BIO-TMT LABOfiATOBBS. tapoat Praparad aad tatad by: D. I. Gordot. D.V4I.. Pb.O. taport Apprmd by; td WATER PCB-SD0000012901 Wifaiaf tlO-TIST DT Ka. iZ2-*T3N I tun > ad knr J m rfBTk J6 Tbsra la srldmia at clMtssl sdhat as A* km vfcMi is at ttai rf ths Mad C2*-MaodO wfillai. lids eMS d a ad in fa* aalaala is bspaSasa r cfastasgiafeipalaam. Tfas kypm-bwpfeic edb ltr| rail--a ad light sMatag qMglMB kkk is psnhably rich b> glytogas and MpIMc Mtalaa. Is asms mill, rtag-ahapsd atrwctons. probably i apraasntlsg whorls ad proUJsratlT* sdoylasdc radniliaa. n asaa Tbs hyparplasdc surrouading calls od bi am tbs saas hypsrtraphic calls Tbesa luwhilaa *Uck ihsnd aUasi ad haps, itaiabag or a doctalar ar ol msta steals, lawalTMiaa. as rare Thsra was sa rrtdaaaca at study. at tfasaa WATER PCB-SD0000012902 VwU. Uxktar. D.T.M..M.S. DtpL, Am. C*tUf* Ttt PuX WATER PCB-SD0000012903 '*. k ' ` -mr Lhrar UatoM Arbiter 13tt IM - Rata SVv loitsa SOS TX T8 TB tmwim ckm / 2/10 1/0 rcl Kyptitnykr 0/0 0/10 0/0 N4mU* HyftfUaU e/a 0/10 0/0 Htyaton* o/t 0/10 0/0 Dwtvlu Kn*iyUtia 0/1 0/10 1/0 CbalMfi** Kt|Mnii e/t 0/10 0/0 Htytwwlhlir Macniii i/a 0/10 0/0 CmmI 2/10 0/10 0/10 0/10 0/10 0/10 0/10 000318 - -'si- WATER PCB-SD0000012904 VmwIm rcil Uyy>HW|ty NoOmUx Hr?** HaftMM Ovcttlii 4n*rpltilt Hp4ucUalu TJ #/10 0/10 0/10 0/10 C/10 0/10 1/10 'C- TS 0/0 o/a o/a o/a o/a o/a o/a rm a/o 0/0 0/0 0/0 0/0 0/0 0/0 Coatrol 1/10 0/10 0/10 0/19 0/10 0/10 1/10 $ WATER PCB-SD0000012905 PllMn Uvar UiltM - Amlat M* : U MMk iKrlflct - Im TX Tiilit Ckoafo 2/1* r*cai H)ymip% 0/10 IMitor lypitpluU 0/10 hum Delu MyyaryUate 0/10 0/10 0/10 HtfitmllaUr Htcmlt 0/10 TB 4/10 0/10 0/10 0/10 1/10 0/10 0/10 -. :;t m 0/0 I/O 0/0 9/0 0/0 0/0 0/10 T 10* C--trol 1/10 0/10 0/10 0. 10 1/10 o/:o 9/10 V* ' . -` . ' ' WATER PCB-SD0000012906 ' >> ' " V' .t'.f Prlauy Um Uiiwi * AtocIot BO Trmlait Skcrlflc* - bti 107 Vuwlir Gain* Txtl Hiiaimylir NoOoUr Hrp*ryUU Ht^uotu DvetaUr HTPrpUiU Cb 1m( la -Htpataata H*p4tcUuUz NcroU t: T/ll 2/11 0/11 0/11 1/11 0/11 1/11 Soar TU nn 0/10 0/20 1/10 0/20 2/10 0/20 0/10 2/20 1/10 0/10 1/20 ` 1/20 1/10 o/:o Control 1/13 0/23 1/21 0 21 1/21 u/21 l 23 WATER PCB-SD0000012907 .c-m | 7 u u jlO O ppn 757" ' ; 7 SS " M4 MH f?1 *4 ^*4 * *4 ? . 5..g.. H ifiij Vocal McrMlt at hapatocytoa Focal IfnykoM laflltrattoM Vocal hypertrophy at hopatocylaa NoAilar hyparplaala at hopatocytaa Vocal Mia h*cl IqrfDi^MU l(|laloma Chalaayia-hapata |<eliculum Coll Sarcoma! malaataltc Mammary tumor, malaalallc Tlanitm lami.i, fiir.il WATER_PCB-SD0000012908 &p"_ ;"*? ''*-.... , * ;. -y-# ABOCfcOR . 1242 TeboUtloo mt ladtri+u.1 Unr Lailoo* la KaU Sacrifice Lata real Doae Animal) Loral No. tad i* 1 Sea $3 2 !*i If bl* HV ]sL k & Moath j C-I . 101114 1012" | 1013" i 10U" I 1015"! 1026F1027" 102S" 1029" 1030" Urn JUilRat ft ti fl ji H !| 1J I s* b h. i 1 * * to T .i ( 'J:. , 3! If: at! I il If! i I C19-Gppm 104134 1042" I 1043" ; 1045" ; iosst I 1057" I Hi Si" ; 1059" 'C.m 107134! 1 OCppm 1072" ! 1073" | 1075" ' tostr 10i7" j 10i" 104" 1090" Control SO 134 02" MS" 04" 05" ur j 117" IIS" 19" 20" ! I WATER_PCB-SD0000012909 jir.-,'v.r "Si- : r |jl : . ufa-^n, , "j: . J..L N-C> -vr-*.? 'W W IMv/Trs4 a m n m ry h ivn o r, tn u U |lc WATER_PCB-SDOOOO012910 L. r 2 #tr? ^5A l i * f" .[! JtftHKtifttHHtfStiHtfttttf * -. * CftipUiaak mwtite . : ^hmnyiii fmlMnwto tl. . : i`,' "P"!l '* . 1' r*ttttyasfcU, . . NmI knuMftiyW bfUMytti . IMdiri^ylHli mt IvfiMiytoi VWI Ml* *tat f H ;< [ 8* V ' f ' V^y. . *,*;s^ r*. . <r'..- ' P;.:- 4ft% V}V,^.^o ?V' wtiM&k&M- I<U...; :A',>*,* . < 1 tr -V^rtsT- * ' . * f.f'H f * Chl|lf tlflhnil. |5- ,.<11v';,.*. -I ' V o u I(liculm Call Iiiwwi mt tails Mminry tumor, IMUlUlk *r-! mi* I iii.i, WATER_PCB-SD0000012911 I 9 n r WATER PCB-SDOOOO012912 4 44 4 ' 1 c .ffl lOO^VB CoatTol >V <**'* c * ir it jTT t* { * * * o 4 4+4 *d *0 *o 4 1 |J4 + *d * * "* w a lirf CfMpUMOlC WntUllN of ktf*lN)tM Focal mmmIi ot Uftltcyiti Focal . kaflUratloas mtFocal fcypocltophy hapotocytaa NcchfUUr finttroBc*fttfibiii* C n1 IFocaltM*piadn4oUc**i* . .> to/ cuu|Ulw(iiHna' /' <Vr . Rcilcahun Coll htwou metastatic Mammary tumor, metastatic |\* \ tlt*t>l | flit l 0003 WATER PCB-SD0000012913 Grading >rfm Minimal In Mrarlly ** . UiU in Mvnrltr 4-t-f Mndamtn in imrily **** Uarfcnd in Mvnrlty P Pmaani. no grada WATER PCB-SD0000012914 ' '*.** -V*. , v* ` ,.-?4 '*'** - \:* .~i CO Lciioo VacuoUr Chang* Focal Hypertrophy Nodular Hyparplasia Hepatoma LXictular Hyparplasia C holangio- Hapa loma Hepatocellular Necrosis. Primary Unr Utloai Aroeier 1242 Its Tarmiul iteriflci - Hat* T1 123 TU 12 Grasp 3 nn 12 3 Caitrel 12 0/31 0/27 7/31 0/31 0/27 2/31 0/31 0/27 0/31 0/31 0/27 0/31 0/31 4/27 3/31 0/31 0/27 0/31 1/31 0/27 1/31 4/30 1/19 1/30 1/30 2/19 3/30 1/30 0/19 2/30 0/30 1/19 0/30 1/30 0/19 3/30 0/30 0/19 0/30 1/30 0/19 1. 30 13/20*' S/21 9/20 1/20 2/21 0/20 2/20 S/21 0/20 0*20 0/21 2/20 20 2/21 3/20 0/20 0/20 1/20 0/20 0/21 0/20 1/23 0/24 0/23 0/24 1/23 0/24 0/23 0/24 S/21 b/24 0/23 0/24 1/21 0/24 1 - First evaluation of original ilUtl 2 - Re*cviIuatian of original slides 1 = Evaluation of additional tiaauo sections Fuur animals positiva oa first evaluation, aot on sscad. WATER PCB-SD0000012915 BIO-TIftT Afr.^ah* fm. Vacuolar Chaago Focal Hypartrophy Nodular Hyparplaala Ductular Hyparplaaia Cholangio-hopatoma Hapainiai Llvor ImUu - AlOCLOft 1242 Tmtatl tocrtflca - Rais " td Ort|taal SUh - ' C-l 0/21 0/21 0/21 C-I 1/1* 2/M 0/M C-l S/21 2/21 S/21 4/27 1/M 2/21 0/27 0/ M 0/21 0/27 0/M 0/21 116 Control 0/24 . 0/24 e/24 i/24 0/24 0/24 rtf WATER PCB-SD0000012916 a I0-T1ST JU 117 .MGJi U4 :i'4m c-r !m 41 j ti | mm 4mm iifiKi mm immrn im* | ,m ; 'Ml * I !i i 1 3 i u:;s * *4: ' . t 1 WATER PCB-SD0000012917 II I I rr I j\ } 4 i I r. j : : (w- 911 1SII*0I WATER PCB-SD0000012918 H i i m rj nli.Miq pa f* r*- >!i :t J rrTa??;/_r/?r ---as m*........."............. i"` IS ii > W *1 c: fJ (, o c iQ I i k m K i m, Ifll'O M WATER PCB-SD0000012919 t? Tj c5 c ! rs WATER PCB-SD0000012920 tuxx I B I T WATER PCB-SD0000012921 DMEH CMC4 CEXTSAl FILES Toxicology Section/St. Louis (CO./CI vjoeptvuqcatio.nj ^ , (' ________ SPECIAL (TYPE OF REPORT) REPORT REPORTNO.: MSL-2003 r to cu H-* g3 3 -- r- tO >- >3- 6w-- C3O CJ H-. H-. G W2 c3-- 3 CO a 2 5- h-. E-- CJ H-. C CJ < >- a 32 at j: a. cj t o<n -- 3 </-i O2 3Z >2* a >-- u ^ f-CC w < 2 fc 3 < >-> >4J 3 3< > 3 C3 co c<- a3 3 e- G< t^o O a CJ 2< 3s HI > CU G 2< G3 hi s Ghi a 333 > 3 in a: . < 2 H-> a C. JOB/PROJECT NO.: DATE: October 14, 1981 TITLE: A REVIEW AND EVALUATION OF CARCINOGENICITY STUDIES IN MICE AND RATS AND MUTAGENICITY STUDIES WITH POLYCHLORINATED BIPHENYLS AUTHORS: George J. Levinskas, PH.D. A3STRACT: This is a review and evaluation of studies which deal with the potential carcinogenicity and mutagenicity of polychlorinated biphenyls (PC3s). It is subdivided into 4 sections: Chronic Rodent Studies, Metabolism Studies, Co-Carcinogenesis Studies and Mutagenicity Studies. A brief summary of Epidemiology Studies is added to complete coverage of the issue of carcinogenicity. DH < h- ro oo(NJ i^ 3 DEPOSITION | EXHIBIT jufvim<.)Cft^47 1 s '<3W w R-toasiat SCM 019651 000332 WATER PCB-SD0000012922 COPY NUMBER 1 - Reports Library, R2C 2 - Reports Library, R2C 3 - Reports Library, R2C 4 - DMEH Library, G2WA 5 - DMEH Library, G2WA 6 - R.T. Berendt, E2ND 7 - J.H. Craddock, A2SA 8 - G.J. Levinskas, G2WF 9 - J.G. Nassif, E2.ND . ' 10 - J.M. Norris, Dow Chemical 11 - R.A. Stohr, B3NJ ABSTRACT ONLY This report has been assigned to you. When it is no longer needed, you are responsible for returning it to: _____________ REPORTS LIBRARY, R2C______________ If you transfer it to anyone else, please let your librarian know, so the records can be changed. R-iaai(C) (Rev. i/aoi SCH 019652 000336 WATER PCB-SD0000012923 INTRODUCTION This is a review and evaluation of studies which deal with the potential carcinogenicity and mutagenicity of polychlorinated biphenyls (PC3s). It is subdivided into 4 sections: Chronic Rodent Studies, Metabolism Studies, Co-Carcinogenesis Studies and Mutagenicity Studies. A brief summary of Epidemiology Studies is added to complete coverage of the issue of carcinogenicity. This review does not discuss the effects of impurities or contaminants, particularly polychlorinated dibenzofurans (PCDF), which are reported to be present in some PCB mixtures (Brinkman and deKok, 1980). The presence and amounts of such impurities have not been specified in the materials used in many studies. Thus, attempts to apportion the observed biological effects between impurities and PCBs would only add further " conjecture to a subject which currently is rife with speculation. By contrast, specific chemical and trade names have been used to identify materials that were studied instead of the all encompassing term PCBs. This was deemed necessary because there are too many one-sided generali zations in the literature. Every adverse finding is, by implication at least, extrapolated to the entire class of materials designated as PCBs. Conversely, every report which has failed to find an adverse effect is careful to stipulate that it relates only to the substance studied. Even more difficult to contend with are misleading references to adverse findings which are not substantiated by the actual publications referred to. An example of this occurred in a discussion of in vitro metabolism studies with liver microsomal enzymes which stated that the formation of PCBmacromolecular adducts had been demonstrated. Among the references ` 1` SCM 019653 000337 WATER PCB-SD0000012924 cited was a paper entitled "The in vitro binding of 2,2',5,5'-tetrachicrobiphenyl metabolites to rat liver microsomal proteins". The latter paper clearly states "There is no clear evidence in the data obtained from the work reported here to substantiate covalent binding; however, it is also not "possible to exclude the nondialyzable radioactivity as being covalently bound". (Hargraves and Allen, 1979). To emphasize the point that dose is important in evaluating safety, test exposure conditions have been described so that the reader can compare them to ambient exposure levels encountered in. the literature and else where. Hoopingarner, et al. (1972) in their studies with Aroclor 1254 on cultured human lymphocytes are among the few authors who acknowledged that "The toxic dose of the chemical was several times greater than is usually found biologically". They used concentrations of 100 ppm of Aroclor 1254 in their studies discussed under Mutagenicity. Chronic Rodent Studies Difficulties in comparing and assessing different studies are compounded not only by problems of histopathologic diagnosis but also by variations in experimental design and animal strain differences. Animal feeding studies cited in the literature as evidence for the carcinogenicity of PCBs and related studies of similar duration have been tabulated for mice in Table 1 and for rats in Table 2. Studies have been grouped by the approximate weight percent of chlorine in the materials tested to facilitate comparisons between products from different manufacturers with the same chlorine content. 2 SCM Q19654 0Q033F WATER PCB-SD0000012925 Mouse studies in Table 1 were conducted with high dietary levels of PCBs. Kimbrough and Linder (1974) reported a 52% mortality for mice fed 300 ppm of Aroclor 1254 for 6 months and then held an additional 5 months. During that interval, control mice had a 32% mortality. This appears to be quite high for control mice about 1 year of age. In a report discussed under Co-carcinogenesis, Kolier (1977) studied mice injected with Aroclor products and Moloney leukemia virus. Some data from his study, although not shown in Table 1, are quite similar to those of Kimbrough and Linder (1974). Kolier (1977) fed diets containing Aroclor 1221, 1242, or 1254 to groups of 25 Balb/c male mice for 6 months. After that time, some mice were sacrificed for examination and others were returned to a control diet for another 3 months and then sacrificed. Dietary levels of each Aroclor were 375, 37.5, or 3.75 ppm. The only feeding regimen that was toxic was 375 ppm of Aroclor 1254. Only 8 mice of that group survived for 6' months, giving a mortality of 68%. Other authors cited in Table 1 (Ito, et al. , 1973a,b and Nagasaki, et al. , 1972, 1974, 1975) did not indicate how many of the mice in their studies died when fed 500 ppm of Kanechlor 500, which has a chlorine content similar to Aroclor 1254. Kolier (1977) reported that "PCBs did not produce hepatic neoplasia".There was marked liver injury in mice fed 375 ppm of Aroclor 1254, mild liver injury which persisted through the 3-month recovery period at 37.5 ppm, and no hepatic lesions at 3.75 ppm. Moderate liver injury was produced by 375 ppm of Aroclor 1242, but this regressed and no hepatic lesions were seen 3 months after mice were returned to a control diet. Other dietary levels of Aroclor 1242 and all levels of 1221 did not produce liver changes. Highly significant mean liver weight increases after 6 months of feeding occurred in all Aroclor 1254 groups and in the group fed 375 ppm of Aroclor 1242. Three months after mice were placed on a -3 SCH 019655 onorw WATER PCB-SD0000012926 control diet, mean liver weights of each group had decreased. The decreases were highly significant for the 37.5 ppm Aroclor 1254 and the 375 ppm Aroclor 1242 groups. These observations are generally consistent with those of other investigators which show regression of lesions when PCQ dosing is ended, the degree of regression depending upon the duration of the recovery period. . Data for mice reported earlier by Nagasaki, et al. (1972) appear to have been included in the later publications of Ito, et al. (1973a,b). It is interesting to note that the tumors were called hepatomas in the former publication and were labeled well-differentiated hepatocellular carcinomas in the latter. Kimbrough and Linder (1974) apparently also felt that these same data on mice were reported in these 2 papers as they reference the . production of hepatomas in male dd mice to the later publication by Ito, . et al. (1973b). Similar terminology troubles beset the 1974 and 1975 Nagasaki publications. Data from the later publication list 9/17 males and 4/17 females fed Kanechlor 500 as having liver tumors. Table 1 shows data from the earlier publication; among males, 9/17 had nodular hyperplasia and 7/17 had hepatocellular carcinoma while the female incidence of 4/17 was described as nodular hyperplasia. The hepatomas described by Kimbrough and Linder (1974) were later referred to as "neo plastic nodules (hematomas [sic], hyperplastic nodules)" by Kimbrough, et al. (1978). The use of the term hepatoma has created confusion. With respect to terminology of tumors in the mouse liver, "Hepatoma is a collective term used to describe the progressive stages of tumour development from the lesion called "hyperplastic nodule" or "simple hyperplastic growth" to the morphologically and biologically malignant neoplasms" (Turusov and Takayama, 1979). Nevertheless, whatever '4* SCH 019656 000340 WATER PCB-SD0000012927 terminology is used to describe them, tumors were attributed only to PCBs containing 52-54 percent chlorine. Lower doses of the 52-54 percent chlorine materials, as well as lower chlorinated materials, were not described as tumorigenic. Table 2 summarizes results of rat studies with PCBs. As in the mouse studies, only some studies with materials having a chlorine content of 52-54 percent, or higher, were reported to produce carcinomas in the livers of rats, and even for those materials this was not a consistent, reproducible observation. Odashima (1976) reported the results from a series of different tests used to evaluate potential carcinogenicity of several compounds, including PCBs. Two tests are of sufficient interest to mention here. One is the transplacental method in which rats were treated for 3 days during pregnancy. These were days 15, 17, and 19 or 14, 16, and 18, depending on the strain used. The total dose was approximately the maximum one that did not cause abortion or early death of the weanlings. In the other, the newborn method, pups were dosed subcutaneously on days 1, 8, 15, and 22 after birth. The maximum dose was one that did not cause early death of over 20% of the animals. The subsequent observation period in each test was limited to one year after birth. For both Kanechlor 300 and Kanechlor 500, the transplacental test was scored negative and the newborn one as equivocal. Both of these tests gave positive results with some known carcinogens such as 4-aminobiphenyl, benzo [a] pyrene, butyl-nitrosourea and N-methyl-N-nitrosourea. SCM 0lt-57 000^.41 WATER PCB-SD0000012928 Objective appraisal of chronic rodent feeding- studies is difficult. Some .of the rodent studies in Table 1 and Table 2 are of the type used to detect potent carcinogens. They use relatively few animals, high doses of the test material, and have a relatively short duration. Studies of that nature have been included in Tables 1 and 2 to illustrate the various diagnoses of the rodent hepatic lesions observed after dosing with PCBs and to aid in the stepwise analysis of the results of all the reported carcinogenicity studies. In any short-term test with a few animals, the chance occurrence of some tumors is a possibility which must be considered. The probability that an event is a chance occurrence is decreased if it is repeatable. Many of the studies in Table 1 did not produce tumors. Those which were reported to ' produce hepatocellular carcinoma apparently were also reported in other publications as producing nodular hyperplasia, or tumors, or hepatomas. Since terminology of tumors and the use of those terms have subjective, i.e., judgemental, elements it may not be valid to make direct comparisons between different studies on the basis of terminology alone. The rodent studies which were of less than lifetime duration raise a question as to whether or not cancers would have occurred had the feeding periods been extended. That question is speculative, and it cannot be answered from the present data. Nevertheless, negative studies even if they are of relatively short duration are part of the overall evidence which has to be considered. As a first step in the analysis of the data, the evidence shows that at a minimum, PCBs are not potent carcinogens to rodents because they do not produce cancers when tested in studies designed to detect potent carcinogens. *6* SCM 019658 WATER PCB-SD0000012929 With respect to further analysis of che mouse studies, greater significance can be attached to the data of Kimbrough and Linder (1974) because their study had a larger number of animals, a longer duration, and a relatively high dietary level of PCS. In those respects, it more closely resembles conventional cancer studies. They did not observe any hepatocellular carcinomas. Thus, it can reasonably be concluded that PCBs are not carcinogenic to mice. Review of the rat data in Table 2 shows that there are 2 studies on PCBs with an average chlorine content of 40% (Levinskas, 1SS1 and Weltman and Norback, 1979) and 3 on PCBs wich an average chlorine content of 52-54% (Levinskas, 1981; NCI, 1973; Wasserman, et al. , 1978). Results of those studies are consistent with respect to the absence of hepatocellular carcinomas. This consistency reasonably suggests a conclusion that PCBs ' wich those chlorine contencs are not carcinogenic. Of the 3 studies wich PCBs having an average chlorine content of 60%, one reported hepatocellular carcinomas (Kimbrough, et al. 1975) and 2 did not (Levinskas, 1981 and Weltman and Norback, 1978). Since Kimbrough, et al. (1975) and Levinskas (1981) both used Lot No. AK-3 of Aroclor 1260, che different conclusions they reached are not related to differences in the test material. In addition to the use of a different strain of rat, Kimbrough, et al. (1975) used a different histologic diagnostic criteria. Kimbrough, et al. (1975) used the criteria for classification of specific hepacoceLlular lesions in rats developed at a National Cancer Institute Workshop (Squire and Levitt, 1975). That workshop recommended that the term "neoplastic nodules" replace so-called "hyperplastic nodules" because "Such nodules are proliferative lesions and -7- SCM 019659 000.143 WATER PCB-SD0000012930 are known co be induced by carcinogens and, ac che least, they indicate an increased probabiity for the development of hepatocellular carcinoma" (Squire and Levitt, 1975). However, Pitot, et al. (1978) in a discussion of stages in the progression of hepatocarcinogenesis in rat liver have noted that "The demonstration of carcinoma cells that appeared to arise within the nodules was also reported (13)1, suggesting the nodule was a precursor to the malignant neoplasm. On the other hand, as was shown by Farber and others, the vast majority of the regenerating or hyper plastic nodules disappeared on removing the animals from the carcinogenic diet. Thus, despite the more recent suggestion that these nodules be termed "neoplastic nodules" (14)2, it is difficult to understand a precursor relationship of the nodule to carcinomas if the existence of the putative precursor is so transient." Further, the use of those criteria for classifying experimental hepatic lesions was considered and rejected by Kimura, et al. (1976) in their studies on the co-carcinogenesis of Kanechlor 400 and 3'-methyl-4-dimethylaminoazobenzene. There is concern' that a presently benign lesion might at some future date or under other circumstances be transformed into a malignant lesion. Kimbrough (1979) apparently had that concern when she stated "Although it has been shown many times that the neoplastic nodules are part of the carcinogenic response they are not always included in the statistical evaluation of bioassays, which may lead to erroneously interpreted results, particularly when they are classified as hyperplastic nodules or "nodular hyperplasia" and when the number of animals studied was small (Carcinogenesis Testing Program, 1977)".3 A similar concern appears to have been behind the statement in a recent review (Anon., 1981) that "hexachlorobiphenyl administered to groups of 50 male and 50 female 8 SCM 019660 000344 WATER PCB-SD0000012931 Sprague Dawley rats ac dietary levels of 0 and 100 ppm for 105 weeks was carcinogenic in female rats, producing an increased incidence of liver hepatocellular carcinomas among the dosed animals (Norback and Weltman, 1980. Personal communication)1'. Contact with Dr. Norback (Ribelin, 1981) revealed that her data were available only as an abstract (Weltman and Norback, 1979), and that the abstract and her oral presentation of the data referred to the lesions as neoplastic nodules, i.e., not distinctly tumorous. These 2 examples illustrate the difficulty in establishing that cancer is not present, and they strongly suggest that the different findings reported by various researchers are a reflection of their orientation, training, and philosophical perspective. The latter is defined as the difficulty in separating what is actually being observed under the microscope, from a concern over what it might have become if the animals had lived longer. Since only an abstract has been published, details on the studies by Weltman and Norback (1979) are limited . Their studies are of particular interest because they observed the sequential development of liver changes over a 2-year period. They noted that hexachlorobiphenyl was more toxic than tetrachlorobiphenyl, and that the more toxic, more highly chlorinated hexachlorobiphenyl produced neoplastic nodules only. Again, the weight of the evidence leads to a reasonable conclusion that the carcinogenicity of biphenyls with an average chlorine content of 60% has not been established. 9 " . SCH 019661 O 00.145 WATER PCB-SD0000012932 Overall, one can, surmise chat the inability to resolve the crucial issue of whether or not a lesion is neoplastic in character was one of the factors which led to the following1 statement from a recent Surgeon General's report: "Science, and society have not yet arrived at a final consensus on the detinition of a carcinogen either in the human population or in experimental animals" (DHHS, 1980). Metabolism Studies Several reviews (Goldstein, 1980; IARC, 1978; Matthews and Kato, 1979; Roberts, et al. , 1978; and Safe, 1980) discuss the metabolism of specific isomers as well as mixtures of PCBs. While there are some exceptions, the following general conclusions can be drawn. Both the degree of chlorination and the positions of the chlorine substituents determine the ease with which PCBs are metabolized. In general, the lower chlorinated ones are metabolized and excreted more readily while the more highly chlorinated materials are stored in fat. The process of metabolism converts the fat soluble PCS into a water soluble hydroxylated derivative which can be excreted in the urine. This metabolism and excretion appears to occur via arene oxide intermediates, and the carcinogenicity of some compounds has been attributed to formation of arene oxide interme diates and their binding to subcellular macromolecules. Chlorination at the 4,4' position blocks the metabolism and excretion of PCBs as illustrated below. SCM 019662 0Q0ri4G WATER PCB-SD0000012933 Studies in rats (Hansell, et al. , 1977) and in mice (Morales and Matthews. 1978, L979) are of interest because they report the metabolism of isomers which were fed to rats for 2 years by Weitman and Norback (1979). Hansell. et al. (1977) gave groups of male rats single intraperitoneal injections of 0.2 nmoles/kg of 2,2',5,5'-tetra- or 2,2\4,4',5,5'hexachlorobiphenyl (TCB and HCB, respectively). They measured changes in hepatic mixed function oxidases and the persistence of the PCB in the livers of animals killed at selected intervals for 35 days after dosing. TCB produced a transient, but significant increase in O-demethylase activity only at day 3 while HCB produced significant increases in O-demethylase and aniline hydroxylase activities within 24-48 hours. Induced enzyme activities by HCB peaked at 4-6 times control activity during days 7-14 and were about 3 times control activity at the end of 35 days. Even though equimolar amounts of each isomer were given, liver ' residues of HCB one day after dosing were about 7 times higher than those of TCB. HCB residues decreased relatively slowly with time while TCB residues were more rapidly eliminated and had almost returned to control values by 35 days. Livers from HCB treated rats had centrolobular necrosis at day 35. They also were significantly increased in size, showed increased amounts of smooth endoplasmic reticulum (SER), and appeared to contain increased numbers of microbodies and reduced amounts of rough endoplasmic reticulum throughout the 35 day interval. By contrast, TCB treated livers were similar to control ones except for occasionally increased aggregates of SER on days 3 and 4. Hansell, et al. (1977) stated "It would be interesting to speculate that.. .2,4,5,2',4',5'hexachlorobiphenyl undergoes direct hydroxylation whereas 2,5,2' ,5'tetrachlorobiphenyl undergoes bio transformation via the arene oxide intermediate, thereby accounting for the different slope for the - 11 - . SCM 019663 000.147 WATER PCB-SD0000012934 elimination curve, of the latter compound." Thus, the metabolism (Hansell, et al. 1977) and feeding1 (Weltman and Morback, 1979) study results lead to somewhat different conclusions. The more potent enzyme inducing, less readily excreted HC3 which appears to resist hydroxylation produces neoplastic nodules in rat livers. By contrast, the more readily excreted TC3, which apparently is excreted via an arene oxide intermediate, does not produce tumors when fed to rats for 2 years. Covalent binding to cellular macromolecules also appears to be greater for the more readily metabolized PCB isomers. Morales and Matthews (1978, 1979) compared the covalent binding of 2 hexachlorobiphenyls; the more readily metabolized 2,2',3,3',6,6'-hexachlorobiphenyl (2,3,6-isomer) and the more slowly metabolized 2,2',4,4',5,5'-hexachlorobiphenyl (2,4,5-isomer). Each PCB, with a radiocarbon label, was given orally to groups of mice at a dosage of 7.28 mg/kg on each of five successive days. Animals were killed 1, 5, and 8 days later. The concentration of each PCB was determined in liver, muscle, and kidney. All tissues had consistently higher concentrations of the less readily metabolized 2,4,5-isomer. The more readily metabolized 2,3,6-isomer showed a consistently greater binding to purified macromolecules. The binding was at least one order of magnitude greater than that seen with the 2,4,5-isomer. Results of animal feeding studies with the 2,3,6-isomer would be of particular interest to determine the degree of correlation, if any, between the carcinogenicity of this isomer and its covalent binding to cellular macromolecules. Taken as a whole, metabolism studies suggest that if PCBs are carcinogenic, it should be those PCBs which are more readily metabolized and excreted, i.e., the lower chlorinated materials. This is in sharp - 12 - SCM 019664 000.^48 WATER PCB-SD0000012935 contrast to results on animal studies in which questions of carcinogenicity arise regarding higher chlorinated materials only. On balance, metabolism studies on the formation of arene oxide intermediates would lead one to expect lower chlorinated materials to show a more pronounced carcinogenic response in animals. Conclusions drawn from metabolism studies are not supported by animal feeding studies. Co-Carcinogenesis Studies The position and degree of chlorination of PCBs also are important in inducing microsomal mixed function monooxygenases (Goldstein, 1980 and Yoshiraura, et al. . 1979). Such induction of microsomal monooxygenases could alter the metabolism of exogenous and endogenous substances in the body, as reported in a variety of studies which have been conducted to determine whether PCBs might be cocarcinogens. These are summarized in Table 3 and discussed in greater detail below. Uchiyama and Chiba (1974) inserted 20-methylcholanthrene impregnated threads into the uteri of virgin mice and fed them diets containing up to 100 ppm of Kanechlor 400 or DDT. Animals were killed at various times and the cervical epithelium was examined for cancerous changes. Kanechlor 400 dosed mice showed no significant changes as compared to the controls, but those receiving 100 ppm of DDT "showed a remarkable tendency towards the induction of cancer." Diets containing benzene hexachloride (BHC) isomers and Kanechlor 500 separately and in various combinations were fed to male mice by Ito, et al. (1973a,b). Concentrations of the BHC isomers ranged from 250 ppm to - 13 SCH 019665 OHO.'?/'.'} WATER PCB-SD0000012936 50 ppm. The amount of Kanechlor 500 was 250 ppm or 100 ppm. Test groups consisted of 20 to 30 animals. When fed alone, only a-BHC at 250 ppm produced a high incidence of noduiar hyperplasia and a moderate incidence of hepacocellular carcinoma. A diet containing 250 ppm each of a-BHC and Kanechlor 500 increased the number of hepatocellular carcinomas. In comparison, combinations of 100 or 50 ppm of a-3HC and 250 ppm of Kanechlor 500 yielded a moderate incidence of nodular hyperplasia and produced only a few hepatocellular carcinomas. There were a few nodular hyperplasias in mice fed 100 ppm each of a-BHC and Kanechlor 500. A mixture of 50 ppm a-BHC and 100 ppm of Kanechlor 500 was without effect. Diets containing 250 ppm or 100 ppm of p-BHC and 250 ppm of Kanechlor 500 produced both nodular hyperplasia and hepatocellular carcinomas at a lower incidence than that seen with comparable diets of a-BHC. No liver nodules were seen with 50 ppm of p-BHC and 250 ppm of Kanechlor 500 or with 100 ppm of each. y-BHC and Kanechlor 500 did not produce liver nodules either singly or in combination. Nagasaki, et al. (1974, 1975) reported a similar series of experiments except that Kanechlor 400 was included. The same concentrations of the PCBs, 250 and 100 ppm, were used. Their test groups consisted of 20 to 38 male mice. For a-BHC and Kanechlor 500, they presented essentially the same data as Ito, et al. (1973a,b). While Kanechlor 400 did not produce liver nodules when fed alone at 250 ppm, there was an increase in the incidence of hepatocellular carcinomas when it was fed in combination with 250 ppm of a-BHC. The increase was similar to that reported by Ito, et al. (1973a,b) for a-BHC and Kanechlor 500. A combination of 100 ppm of a-BHC and 250 ppm of Kanechlor 400 produced only a few nodular hyperplasias and 100 ppm of each in the diet did not induce any liver nodules. - 14 SCM 019666 OQQ.i^ WATER PCB-SD0000012937 The effaces of PCBs on cumor induction by diethylnitrosamine (DEN) have been studied extensively. Male rats were given 25 ppm of DEN in their drinking water and 500 ppm of Kanechlor 500 in their diet concurrently for 20 weeks, returned to a control diet for 4 weeks, and then killed (Makiura. et al. 1974). Neither liver tumors nor nodular hyperplasia were seen, even though rats receiving the same DEN treatment alone had a 92% incidence of liver cancer. The livers of rats treated with Kanechlor 500 only also had no tumors. When administered after DEN, Kanechlor 500 markedly enhanced liver tumor production as reported in a series of papers by Nishizumi (1976, 1979a, 1979b). He described studies in which male rats were given 50 ppm DEN in drinking water for 2 to 10 weeks, after which they were intubated twice weekly with a com oil solution of Kanechlor 500 for 6 to 12 weeks. Kanechlor 500 doses were either 0.2 ml of a 5% or 0.1 ml of a 10% solution. Since experiments were terminated at 20 or 52 weeks after dosing, some animals received untreated diets prior to sacrifice. Only one paper (Nishizumi, 1976) contained results for animals dosed only with Kanechlor 500 alone. That treatment produced enlargement of the livers of rats but no neoplastic lesions. After pre treatment with DEN, however, hepatocarcinogenesis was enhanced as evidenced by the earlier appearance and a significant increase in the number of tumors. A similar dosing pattern was used by Preston, et al. (1981). Male rats were given drinking water containing 66 pg DEN/ml (66 ppm) for 5 weeks after which they were fed for 18 weeks on a control diet or one containing 100 ppm of either of 2 Aroclor 1254 diets. One was Aroclor 1254 as received. The other was an Aroclor 1254 which the authors claimed to have purified by removing polychlorinated dibenzofuran (PCDF) impurities by adsorption onto and subsequent elution from activated Florisil. No analysis was made of Aroclor 1254 as received for " 15 " . SCM 019667 000332 WATER PCB-SD0000012938 PCDF. The authors reported recovery of PCDF at a level comparable to 30 pg of tetrachlorodibenzofuran (TCDF) per 10 g of Aroclor 1254. TCDF was used as a positive control and a standard for quantification for the purification process. Other animals received one of the 2 Aroclor 1254 diets only during the latter 18-week interval. ' No evidence of hepatic tumor formation was seen in control animals or those receiving 100 ppm of either Aroclor 1254 diet. However, using the diagnostic criteria of Squire and Levitt (1975), they reported that both Aroclor 1254 diets resulted in significantly greater incidences of hepatocellular carcinomas in rats pretreated with DEN as compared to those dosed with DEN alone. Thus, they concluded that the hepatic tumor-promoting ability appears to reside in Aroclor 1254 itself. As the authors of this paper pointed out, it cannot be concluded that PCDFs are not promoters of hepatocarcinogenesis since appropriate studies have not been done on PCDFs alone. Similarly, their findings do not invalidate the hypothesis that some of the other effects reported for some PCBs may have been due to PCDF impurities. The dependence of the inhibition or promotion effect of PCBs on DEN-induced tumors on the dosing sequence was illustrated in a different manner by Nishizumi (1980). Pregnant female rats were given oral doses of 200 mg/kg or 50 mg/kg Kanechlor 500 on days 5, 10, and 15 of gestation and were allowed to deliver their pups. At 28 days of age, pups were given 50 ppm of DEN in their drinking water for 5 weeks. Groups of these pups were sacrificed at 16, 20, and 24 weeks after the start of DEN dosing and their livers were examined. The number of liver tumors in the offspring from Kanechlor 500 treated dams was significantly decreased as compared to the controls. The decreases were more pronounced in males. This decrease in DEN-induced tumors apparently resulted from induction of - 16 SCH 019668 WATER PCB-SD0000012939 microsomal enzymes in the livers of the pups. Those livers contained Kanechlor 500 residues and electron microscopy showed an increase in the smooth endoplasmic reticulum of hepatic cells. Rainbow trout were fed 6 ppb of aflatoxin Bi (AFB!), or 100 ppm of Aroclor 1254, or a combination of both for a year (Hendricks, et al. , 1977). At intervals during' the year, some fish were killed. The remainder were killed at the end of that time. All livers were examined grossly and microscopically. There were no tumors in those fed a control diet or Aroclor 1254 alone, and growth was not affected by 100 ppm of Aroclor 1254. The number of tumor-bearing fish in the group fed AFB t, plus Aroclor 1254 was significantly reduced (to less than one-half) when compared to those fed AFB!, alone. There also were fewer tumors per liver and the tumors were smaller in those fed the mixture. ' In the studies cited earlier, Makiura, et al. (1974) also fed combinations of 500 ppm Kanechlor 500 with 300 ppm of 3'-methyl-4-dimethyIaminoazobenzene (3'-Me-DAB) or 150 ppm of N-2-fluorenylacetamide (2-FAA). Results similar to those when PCBs were fed concurrently with DEN were obtained. Kanechlor 500 reduced the incidence of liver tumors to zero from 65% for those treated with 3'-Me-DAB and from 54% for those treated with 2-FAA. However, the effect of PCBs on the tumorigenicity of 3'-Me-DAB appears to depend on the dosing sequence as shown for DEN. Kimura, et al. (1976) used different sequences to feed groups of rats diets containing 400 ppm of Kanechlor 400 for six months and 600 ppm of 3'-Me-DAB for 2 months. Some received Kanechlor 400 alone and some only 3'-Me-DAB. Others received one diet, then the other, after a 2-month interval on control diet. A final group received Kanechlor 400 for - 17 - SCM 019669 WATER PCB-SD0000012940 4 months and both materials for another 2 months. No hepacocarcinomas were produced by Kanechlor 400 alone or when it was given before or during the overlap with 3'-Me-DAB. No hepatocarcinomas occurred in control animals. The incidence of liver cancer rose from 13% in those receiving 3'-Me-D AB alone to 64% in the group which received Kanechlor 400 after 3'-Me-DAB. The inhibitory effect of Kanechlor 500 on rat liver tumors was evident when rats were dosed with two carcinogens (Makiura, et al., 1974). Combinations of 3'-Me-DAB and DEN or 2-FAA and DEN were administered with and without Kanechlor 500 at the concentrations stated earlier. The liver cancer incidence fell from 92% to 8% when Kanechlor 500 was given to rats dosed with 3'-Me-DAB and DEN. It went to zero from 82% for those receiving 2-FAA and DEN. Nishizumi (1979a,b) studied the effects of various combinations of DDT and sodium phenobarbetal (SPB), in the absence and in the presence of Kanechlor 500, on the induction of rat liver hepatocellular carcinoma by DEN. Pretreatment of rats with DEN followed by DDT, by SPB, or by a combination of both produced low incidences of liver cancer while DEN pretreatment alone did not induce cancer. When Kanechlor 500 was included with each of the preceding dosing regimens, there was a marked increase in liver cancers. Increases resulting from joint administration of compounds after pretreatment with DEN were lower than those observed for DEN followed by Kanechlor 500 alone discussed earlier. Ito, et al. (1978) fed diets containing 200 ppm of 2-FAA to male rats for two weeks and then a diet containing 1000 ppm of an unspecified Kanechlor - 18 SCM 019670 000354 WATER PCB-SD0000012941 mixture for 8 weeks. Partial hepatectomies (PH) were performed on some rats during- the third week of the study. Feeding of 2-FAA was without effect, but that diet plus PH produced a few hyperplastic nodules in the Liver. Treatment with both diets caused an even greater incidence of hyperplastic nodules and both diets combined with PH produced a marked rise in the number of liver nodules. DiGiovanni, et al. (1977) and Berry, et al. (1978, 1979) studied Aroclor 1254 in a two-stage mouse skin carcinogenesis assay to see if it was a tumor initiator or promoter. The shaved skin of female mice was dosed with Aroclor 1254 at a level of 100 pg or 625 pg per mouse. This was applied alone as an initiator or from 5 minutes to 72 hours before initiation with 7,12-dimethylbenz(a]anchracene (DMBA). One week later, mice received twice weekly applications of 5 pg of the phorbol diester' promoter 12-0-tetradecanoylphorbol-13-acetate (TPA) for 32 weeks. Animals were observed for both papillomas and carcinomas. Aroclor 1254 alone produced a few papillomas. The authors of these reports apparently faced a dilemma- common to scientists, i.e., how much significance should be attached to the observation of a low incidence finding which may or may not have a causal relationship to treatment. On the basis of these few papillomas, Aroclor 1254 was called a "weak tumor initiator" (DiGiovanni, 1977). Later, it was described as possessing "little or no tumor-initiating properties" (Berry, et al. , 1979). While Aroclor 1254 had a negligible effect on tumor induction when given 5 minutes before an initiating dose of DMBA, it markedly inhibited tumor induction when given 18 to 72 hours before DMBA. In other studies, an initiating dose of 200 nmole of DMBA was applied to the shaved backs of mice. After one week, 100 pg of Aroclor 1254 was applied twice weekly for 30 weeks. Aroclor 1254 failed to - 19 - SC* 019671 000.755 t, WATER PCB-SD0000012942 promote tumors while 0.2 yg doses of TPA applied in a similar manner induced an average of 8 papillomas per mouse. The authors concluded that Aroclor 1254 possessed little or no tumor-initiating or tumor-promoting properties. . The transplantability and growth of Walker 256 carcinosarcoma in rats was inhibited by Aroclor 1254 (Kerkvliet and Kimeldorf, 1977a,b). Diets . containing up to 800 ppm of Aroclor 1254 were fed to rats of both sexes for 30 days, after which they were given intramuscular injections of Walker tumor cells. Nine days later, during which time they continued to receive Aroclor 1254 in their diets, animals were killed and the tumors were dissected out and weighed. Mean tumor weights of all Aroclor 1254 fed groups were significantly reduced as compared to their sex-matched controls, and the reductions were dose-related to the dietary level of ' Aroclor 1254. Body weight gain was depressed in males receiving 400 ppm or more of Aroclor 1254. Females showed reduced body weight at 100 ppm, the lowest level fed to that sex. Intraperitoneal injections of 50 to 200 mg/kg every other day for 14 days after injection of a small inoculum of Walker 256 cells (103) inhibited both the development and growth of the Walker tumor. The number of tumor takes and the size of the tumors were reduced and the tumor latency period was increased. Body weight gain was reduced only at the 200 mg/kg dosage. Alternate day intraperitoneal injections of 100 mg/kg and 200 mg/kg of Aroclor 1254 after a large inoculum of Walker 256 cells (107) were continued for 60 days. Control animals receiving tumor cells only had 100% mortality by day 20 with a mean survival time of 13.5 days. Mean survival times were significantly increased to 17.7 days and 18.9 days for animals dosed with 100 mg/kg and 200 mg/kg of Aroclor 1254. A few animals survived to 60 " 20 " SCM 019672 OOQ35C WATER PCB-SD0000012943 days, and four receiving Che higher dosage of Aroclor 1254 showed cocal regression of their bilateral tumors. Tumor growth rates in Aroclor-treated animals were significantly reduced. Again, body weighc gain was depressed only at Che 200 mg/kg dosage. An experiment was undertaken to determine if there was an optimum time period for the antitumor effect of Aroclor 1254. Intraperitoneal injections of 100 mg/kg of Aroclor 1254 were given on 5 consecutive days before tumor inoculacion, on 5 days after inoculation, and daily from 5 days before until 10 days after inoculacion. This study was ended 14 days after Che initiating tumor inoculum was given. While all dosing schedules reduced tumor growth, there were variations in the responses. The greatest inhibition of tumor growth resulted from dosing animals for 15 days. Almost equally effective were precreatment and treatment starting with inoculation of the Walker cells. The delayed treatment starting after 5 days was least effective as Che tumor had become established. While early treatment reduced tumor growth, it was less effective in preventing metastases and death of the animals. - Kerkvliet and Roller (1980) offered groups of male mice diets containing 10, 100, or 500 ppm of Aroclor 1254 for 15 weeks prior to injecting them with MSB, an established tissue culture cell line derived from Moloney sarcoma virus. They measured tumor growth and cell-mediated toxicity over the next 19 days. A dietary level of 500 ppm of Aroclor 1254 resulted in marked weight loss and deaths. This was reduced to 250 ppm at 11 weeks, and a few weeks later surviving animals were placed on a control diet. They reported that Aroclor 1254 pretreatment enhanced the growth rate of the MSB tumor and inhibited or delayed the development of cellular immunity against the tumor. SCM 019673 000357 WATER PCB-SD0000012944 KoUer (1977) fed groups of mice diets containing 3.75, 37.5, or 375 ppm of Arocior 1221, Aroclor 1242, or Aroclor 1254 for 6 months. About 2 weeks after being placed on test diets, some mice in each group were inoculated intraperitoneally with Moloney leukemia virus. None of the dietary levels of -any Aroclor affected, i.e., did not promote or induce, the oncogenesis of Moloney leukemia virus. Since PCBs are enzyme inducers, their biological effects resemble those of other enzyme inducers. Peraino, et al. (1978) noted that phenobarbital had a specific tumorigenic enhancing effect that was restricted to the liver. With the exception of the mouse skin painting studies and those in which tumor cells or a virus were injected, all of the studies in Table 3 were concerned with liver tumors. Peraino, et al. (1978) also pointed out resemblances between phenobarbital and PCBs. Both substances "enhanced hepatic tumorigenesis" when given after DEN, and both "exerted a protec tive effect against hepatic tumorigenesis" when administered concurrently with AAF or DEN. The comparison between PCBs and phenobarbital is extended by the observation of Berry, et al. (1978, 1979) that PCBs did not promote skin tumors in mice pretreated with DMBA. Peraino, et al. (1978) refer to a study4 in which skin tumorigenesis was not enhanced in mice fed phenobarbital after skin painting with DMBA. Lichti, et al. (1978) described the induction of ornithine decarboxylase (ODC) in mouse epidermal cell cultures by TPA. Aroclor 1254, apparently at much higher concentrations than TPA, induced a low but reproducible stimulation of ODC in the same system. They also noted a report5 that a high intraperitoneal dose of Aroclor 1254 induced ODC in the liver of rats. After commenting that PCBs "warrant further investigation into their - 22 - SCM 019674 00035r WATER PCB-SD0000012945 possible action as tumor promoters," they added that "These compounds are being tested on carcinogen-initiated mouse skin (T.J. Slaga, personal communication)". The series of publications by Berry, et al. (1978, 1979) and DiGiovanni. et al. (1977) show that PCBs are not promoters on mouse skin. Authors of some of the mutagenicity studies to be discussed (Danz and Urban, 1980; Norback, et al. , 1981; and Stadnicki, et al., 1979) have suggested that PCBs may act as promoters of carcinogenicity. In general, those comments appear to have been offered as hypotheses to aid in understanding the observations which had been made. This also appears to be the case for the NCI study on Aroclor 1254 which was reviewed by the Data Evaluation/Risk Assessment Subgroup of the Clearinghouse on Environmental Carcinogens. That group, charged with the responsibility of providing a peer review of NCI bioassay reports on chemicals studied for carcinogenicity, made and accepted a motion to add the following to the report summary: "Based on the liver proliferative lesions in the treated rats and published reports, it is suggested that Aroclor 1254 may be a tumor promoter" (NCI, 1978). With respect to tumor promotion, Weisburger and Williams (1980) state that "certain inducers of liver metabolic enzyme systems, such as phenobarbital, DDT and BHT, when administered after minimal doses of primary hepatocarcinogens exerted a powerful promoting effect". As mentioned earlier, PCBs also induce liver metabolic enzyme systems. However, since the carcinogen alone produced tumors in animals in the co-carcinogenesis studies summarized in. Table 3, it appears that the doses were greater than minimal. In addition, even though the co-carcinogenesis studies were of ' 23 " SCM 019675 0nQ"59 WATER PCB-SD0000012946 shorter duracian, several used much higher dietary levels of PCBs than those fed to rats for 2 years. The latter studies are the ones which gave rise to questions of carcinogenicity. Thus, while the evidence indicates that PC3s are not carcinogenic, their reported effects in rodent livers following prolonged exposure in conjunction with an initiator may be promotion or inhibition. ' When administered to animals together with a biological agent, PCBs show a promoting or inhibitory effect similar to that seen with, chemical agents. Pretreatment with PCBs inhibited the growth of Walker 256 carcinosarcomas in rats and increased their survival time (Kerkvliet and Kimmeldorf, 1977a,b), enhanced the growth of MSB tumor in mice (Kerkvliet and Roller, 1978) and neither promoted.nor induced the oncogenesis of Moloney leukemia virus in mice (Roller, 1977). ` Mutagenicity Studies Mutagenicity testing of PCBs has ranged from bacterial systems to intact animal studies. A series of studies with eleven bacterial test strains (Heddle and Bruce, 1977; Hsia, et al. 1978; McMahon, et al. 1979; Odashima, 1976; Probst, et al. , 1981; Sugimura, et al. , 1976; and Wyndhara, et al. , 1976) are summarized in Table 4. Hsia, et al. (1973) used both phenobarbital and Aroclor 1254 to induce liver enzymes in rats for preparation of S-9 activation systems. They also tested 4-hydroxy2,2',5,5l-tetrachlorobiphenyl and 2,2',5,5'-tetrachlorobiphenyl-3,4-oxide, a known and a presumed metabolite of 2,2' ,5,5'-tetrachlorobiphenyl, respectively. All three materials gave negative responses with each activation system. Odashima (1976) presented data in tabular form from a series of screening tests. In addition to the results shown in Table 9, " 24 . $CH 019676 0003G(i WATER PCB-SD0000012947 bacterial strains W'?2, TA100, TA98, H-17, M-45, W3110, and TA1973 are shown in groups in their Table 5 with a text notation that not all chemicals were tested with each strain. For Kanechlor 300 and 500, such of the preceding strains as were tested showed a negative response. The group consisting of H-17 & M-45, WP2 try" (her'' & her") shows a plus sign for Kanechlor 300. Presumably, one or more of those bacterial strains gave a positive response. Kanechlor 500 was listed as giving a negative response with the latter group. With the exception of Wyndham, et al., (1976), all of the studies were negative with and without the addition of a microsomal enzyme activation system. It is somewhat difficult to reconcile the text and the graphs in the publication by the latter authors. They stated that their "results clearly showed that as the degree of chlorination decreased themutagenicity increased, a concentration of 100 pg of 4-chlorobiphenyl in the test medium gave over 2000 revertant colonies per plate. The higher chlorinated biphenyls show very little activity as mutagens". While Figure 3 in their article shows their marked increase in revertants for 4-chlorobiphenyl, the same figure also shows results for Aroclor 1221, which averages 1.15 chlorine units per molecule. The mutagenicity of Aroclor 1221 is not discussed in the text, but Figure 3 shows that at a concentration of 100 pg per plate, it produced about one-tenth of the revertant colonies that 4-chlorobiphenyl did. Thus, a 15% increase in chlorine content produced a 10-fold decrease in the reported mutagenic activity. In light of that sharp reduction of mutagenic activity with a slight chlorine increase, it is difficult to understand the statement in their text that Aroclor 1254 "was only weakly mutagenic." Results from Aroclor 1254 (average of 4.96 chlorine per molecule) are not shown in their ' 25 ' . SCH 019677 000362 WATER PCB-SD0000012948 Table 3, but 2,2' ,5, 5!-tetrachlorobiphenyl (average of 4 chlorine per molecule) was presented. At 100 pg per plate, 2,2',5,5'-tetr3chlorobiphenyl was virtually devoid of mutagenic activity. Overall, primarily because McMahon, et al. (1979) reported that 4-chlorobiphenyl was not mutagenic, it can only be concluded that the findings of Wyndham, et al. (1976) are aberrant. A similar conclusion that the findings reported by Wyndham, et al. (1976) are unfounded because attempts to repeat them have been unsuccessful was reached by the State of California (Anon. 1981). In a variant of this test, Stott and Sinnhuber (1978) used Aroclor 1221, 1242, 1254, and 1260 to induce the mixed function oxidase system of the liver in trout. The subraitochondrial fraction of those livers was used to activate the metabolism of AFBi which was assayed for mutagenicity using strain TA 1538 of S. typhimurium. The mutagenic response was decreased compared to the concurrent control using untreated trout liver. A general pattern of decreasing response with increasing degree of chlorination was observed, except for Aroclor 1260. Induction of mutagen detoxifying enzyme systems was suggested as a possible explanation for the apparent conflict between the reported induction of trout mixed function oxidases and the decrease in mutagenic activity. These results are consistent with those of Hendricks, et al. (1977) discussed earlier who reported that Aroclor 1254 reduced hepatic tumors in trout fed AFBX. On the basis of slower sedimentation rates in alkaline sucrose gradients, Stadnicki, et al. (1979) reported that 2,2`,5,5'-tetrachlorobiphenyl (TCB), a mixture of the 3-hydroxy and 4-hydroxy derivatives of TCB and the 3,4-epoxide of TCB induced single strand breaks in DMA of L-929 cells. - 26 SCH 019678 000361- WATER PCB-SD0000012949 At 100 pg/ml. each of che three materials caused all of the DMA co come out in fractions at Che cop of che gradient. The epoxide caused some breakage down Co 1 pg/ml. The mixture of hydroxy derivatives caused significant breakage at 20 pg/ml and only slight breakage at 1 and 10 pg/ml. TC3, che lease pocent, caused lesser breakage at 20 pg/ml and was without effect at 1 and 10 pg/ml. Nilsson and Ramel (1974) conducted genetic tests on adults and larvae of Drosophila melanogaster fed Clophen 30 and Clophen 50. These PCB mLxtures were wichout effect on the loss of sex chromosomes used Co measure chromosome breaking action and nondisjunction of the sex chromosomes. Tazima (1980) has described a specific locus test using the silkworm (Bombvx mori). Neither Kanechlor 300 nor Kanechlor 500 showed mutagenic activity in that system. ' Hoopingarner, et al, (1972) induced mitosis in cultured human lymphocytes with phytohemagglutinin and treated them with 100 ppm of Aroclor 1254. There was no effect on the mitotic index, satellite association, chromatid gaps or chromatid breaks in comparison to control human lymphocytes. Odashima (1976) also studied the incidence of chromosomal aberrations. Their in vitro test used Yoshida ascites sarcoma cells cultured for 6 to 72 hours in the presence of PCBs at concentrations producing a minimal or 50% growth inhibition. Kanechlor 300 gave a positive response and Kanechlor 500 a negative one. For an in vivo system, they examined bone marrow cells 6 to 48 hours after adult or newborn animals were given an approximately lethal dose of PCBs. This test gave the opposite result; Kanechlor 500 was positive and Kanechlor 300 was negative. They noted " 27 " SCH 019679 WATER PCB-SD0000012950 that chromosomal aberrations frequently occurred in controls and that chemicals were called positive when they induced more than twice the aberrations in controLs. In "a Syrian hamster cell transformation system (Pienta, 1980), Aroclor 1254 was one of several chemicals tested double-blind. It gave a negative result. Mouse embryo fibroblasts also have been exposed to different PCBs. Nesnow, et al. (1981) studied the cocarcinogenic action of agents which increase microsomal mixed-function oxidase activity in the C3H10Tl-sCL3 transformation assay. After a 48-hour pretreatment with Aroclor 1254, cells were then treated with benzo(a)pyrene [B(a)P] and the agent for an additional 24 hours. Aroclor 1254 did not increase B(a)P-mediated transformation and no Type II or Type III foci were observed. Norback, et al. (1979, 1980, 1981) exposed CSHIOT^ cells continuously for 6 weeks to 10 pg Aroclor 1254 per ml of medium. Treated cells developed Type III foci. Cells exposed to the same concentration of Aroclor 1254 for 24 hours or to 1 pg of Aroclor 1254 per ml of medium did not develop Type III foci. Continuous exposure of cells to Aroclor 1260 and 2,4,5,2',4',5'-hexachlorobiphenyl also caused formation of Type III foci. A clone from a focus transformed by Aroclor 1254 induced sar coma formation when inoculated in irradiated mice. Foci from Aroclor 1260 and 2,4,5,2',4',5'-hexachlorobiphenyl had not been characterized further. Since transformation occurred only after continuous exposure, Norback, et al. (1981) suggested that the effects of PCBs in culture include promotion. When fed to rats, however, 2,4,5,2*,4*,5'-hexachlorobiphenyl produced neoplastic liver nodules, not hepatocarcinomas (Weltman and Norback, 1979). - 28 SCM 019680 0003b* WATER PCB-SD0000012951 Wong, et al. (1979) claimed that 4-chlorobiphenyl induced an increase in unscheduled DN'A synthesis in Chinese hamster ovary cell cultures in the presence of hydroxyurea, a chemical agent which suppresses normal replicacive DN'A synthesis. Few details were presented regarding the validation and reliability of their test system. By contrast, Aroclor 1254 gave a negative response in an unscheduled DNA synthesis in primary cultures of adult rat hepatocytes (Probst, et al.. 1981). The latter authors presented results of an extended series of compounds tested in their system. Danz and Urban (1980) have proposed using the mitogenic response of the rat adrenal cortex after dosing the animals with a test substance as a short-term test for evaluating the promoting action of chemical compounds. Clophen A60, Delor 103s, Delor 106s, and Phenoclor DP6 all gave positive responses in their test system. At 3-6 days of incubation, embryos from ring doves (Streptopelia risoria) fed a control diet or one containing 10 ppm of Aroclor 1254 were examined for cytogenetic changes (Peakall, et al., 1972). The relative frequencies of chromosome aberrations in the . 8 largest chromosome pairs in metaphase cells of allantoic sac and limb bud origin were scored. The 6 control embryos had a mean aberration rate of 0.8% (0-2.0). The aberration rate in 17 embryos from Aroclor 1254 treated birds was 1.8% (0-9.4). In the latter group, there was one chromosome rearrangement, 13 embryos with aberration rates exceeding the mean control rate, and 4 embryos which exeeded the highest control rate. Aroclor 1242 was injected into fertile White Leghorn eggs to give estimated final concentrations of 10 or 20 ppm (Blazak and Marcun, 1975). After 4 or 5 days of incubation, eggs were - 29 SCM 019681 000365 WATER PCB-SD0000012952 injected with coicemide, incubated an additional 45-60 minutes, and embryos were harvested for examination. There was a high degree of early embryonic death, and a few live embryos showed drastically retarded development without malformation. The first five pairs of chromosomes, constituting over 50% of the chromatin material per cell, were examined for detection of clastogenesis. There were no chromosomal aberrations. Heddle and Bruce (1977) injected mice with Aroclor 1254 for five consecu tive days and then examined bone marrow preparations for chromosomal breakage and sperm ceE preparations for sperm with abnormally shaped heads. While nonspecific factors can induce sperm abnormalities, the authors believe the latter also can result from point mutations or small deletions. Aroclor 1254 was judged to be neither carcinogenic or mutagenic. ' Dikshith, et al. (1975) gave male rats oral dosages of 50 mg/kg of Aroclor 1254 on each of 7 consecutive days. Animals were killed at intervals over the next 3 days. Those scheduled for cytogenetic analysis were injected with colchicine 2 hours before killing, after which time the seminiferous tubules were prepared for such study. At autopsy of the other animals, testis, epididymis, and liver weight were recorded and sections were prepared for microscopic study and histochemical determi nations. Livers were markedly enlarged and showed a significant increase in weight compared to controls. There were no differences in body weight or weight and appearance of testis, epididymis, or vas deferens between control and treated rats. Aroclor 1254 treated rats showed a few meta phase figures with abnormal chromosomes whch appeared to be sporadic and not specific to any one type. Histological examination showed testis - 3JU0 - SCH 019682 00Q36o WATER PCB-SD0000012953 and epididymis from Aroclor L2S4 treated rats were comparable to controls although interstitial cells were increased in Aroclor 1254 dosed rats. These cells showed an increase in acid phosphatase activity. The authors concluded that they "found no evidence to suggest that Aroclor 1254 causes significant chromosome damage or histopathological changes in the rat testis." ' Similar studies were conducted by Green, et al. (1973, 1975a) who gave male rats single oral dosages of 5000 mg/kg, 2500 mg/kg or 1250 mg/kg or four successive daily dosages of 500 mg/kg of Aroclor 1242. Other rats received five consecutive daily dosages of 300 mg/kg, 150 mg/kg or 75 mg/kg of Aroclor 1254. Animals were injected with colcemide 3 or 4 hours before sacrifice which occurred about 24 hours after the single dose ` or the series of doses. There were some body weight losses and some ' deaths occurred. Bone marrow from rats dosed with both PCB mixtures and spermatogonial preparations from Aroclor 1242 treated rats were prepared for cytogenetic study. Repeated dosages of 150 mg/kg and 300 mg/kg of Aroclor 1254 produced decreases in the number of mitoses in bone marrow cells. Repeated dosages of 75 mg/kg of Aroclor 1254 and all dosages of Aroclor 1242 were without effect. Neither PCB mixture at any dosage produced a significant number of chromosomal abnormalities in bone marrow cells. At the lower dosages, Aroclor 1242 did not affect mitoses of spermatogonial cells, but 5000 mg/kg or 4 dosages of 500 mg/kg significantly reduced the rate of cell division. No dosage of Aroclor 1242 produced cytogenetic abnormalities in spermatogonial cells. The authors concluded from their studies that Aroclor 1242 and Aroclor 1254 did not possess mutagenic potential. Garthoff, et al. (1977) fed male rats diets containing 5, 50, or 500 ppm of Aroclor 1254 for 5 weeks, after which they " 31 " SCM 019683 000367 WATER PCB-SD0000012954 examined bone marrow and testis samples. There was no significant difference between test and control animals with respect to the incidence of chromosomal abnormalities and the number of cells in mitosis from bone marrow and spermatogonial cells. In a dominant lethal study (Keplinger, et al.. 1972 and Calandra, 1976), albino mice were given single intraperitoneal dosages of 500 or 1000 mg/kg of Aroclor 1242, Aroclor 1254, or Aroclor 1260 and mated on successive weeks to virgin females. There was no evidence of mutagenic effects. Although details on their studies are limited, their results are in general agreement with those from another dominant lethal study in rats conducted by Green, et al. (1975b) with Aroclor 1242 and Aroclor 1254. The former was administered orally at a single dosage of 625, 1250, or 2500 mg/kg or in five daily dosages of 125 or 250 mg/kg. Aroclor 1254 was given orally in five daily dosages of 75, 150, or 300 mg/kg. After dosing, they were mated with untreated females for 10-11 weeks. Another group of rats was given 150 mg/kg of Aroclor 1254 for five successive days and starved overnighc before admittance to females. Other male rats were offered diets containing 25 or 100 ppm of Aroclor 1254 for 70 days, then mated with untreated females for one week. TEM (triethylenemelamine) was used as a positive Control. There was a body weight loss and some deaths occurred in a few groups. Neither the oral dosing nor the dietary feeding of Aroclor 1242 or Aroclor 1254 had any effect on the number of implantations or the number of dead implantations per pregnant female while TEM produced significant postimplantation losses in weeks 2, 3, and 4. These authors noted that the reduction in the number of dividing spermatogonial cells reported earlier for Aroclor 1242 (Green, et al., 1973, 1975a) did not impair the reproductive performance of male animals. - 32 SCH 019684 (V^ "6^ WATER PCB-SD0000012955 Review of the various mucagenicity tests and their results prompts three comments. Chlorinated hydrocarbons do not generally give positive results in Salmonella strains. Therefore, the mutagenic responses with TA1538 (Wyndham, et al. 1976) are either aberrant or else the response is, indeed, Limited to essentially monochlorobiphenyl. The degree of correlation between in vitro mutagenicity tests and carcinogenicity depends upon the initial classification of the test materials. Pienta (1980) classifies Aroclor 1254 as a non-carcinogen. Rinkus and Legator (1980) regard Aroclor 1254 and Kanechlor 500 as having known or suspected carcinogenic activity. Odashima (1976) and Sugimura, et al. (1976, 1977) consider Kanechlor 500 to be carcinogenic and Kanechlor 300 to be non-carcinogenic. Apart from the difficulties they present for correlation, these different opinions about the carcinogenicity of PCBs are a reflection of what data these individuals considered and how they evaluated those data. Finally, in light of the large number of tests conducted to evaluate various mutagenic parameters, it is not surprising that an occasional suspicious or positive finding resulted, simply on a statistical basis. Those occurred in in vitro systems. In vivo tests gave negative results and provide a basis for concluding that ambient levels of PCBs do not present a mutagenic risk. Epidemiology' Studies With respect to epidemiologic studies, it should be noted that there are frequent references in the literature to an epidemiologic study of workers exposed to Aroclor 1254 (Bahn, et al. , 1976, 1977). Those articles are cited in two recent reviews. One states "The epidemiological data provide suggestive evidence of a relationship between exposure to polychlorinated biphenyls and the development of malignant melanoma. ...for practical - 33 SCM 019685 000364 I WATER PCB-SD0000012956 purposes, polychlorinated biphenyls should be regarded if they were carcinogenic to humans" (IARC, 1978). The ocher states "No conclusive evidence has chus far been reported which demonstrates that occupational exposure to PCBs has caused an increased incidence of cancer" (Kimbrough, 1930). The latter author then proceeds to discuss the study by Bahn, et al. (1976). Among the references for these 2 reviews are NIOSH (1978) which has the following comment on the Bahn study, "PCB exposure histories were based on recollections of two company employees. Exposures to other chemicals could not be ascertained.------ To correct these deficiencies in the preliminary study, a more intensive investigation is being conducted (B.N. Kightlinger, written communication, November 1976). A substantial change has occurred in the cohort since release of the preliminary report by Bahn and her coworkers, and it seems likely . that the findings on this new cohort will differ significantly from those of the preliminary study. The final report is not yet available." Recently, Gaffey (1981) reviewed and evaluated existing reports concern ing health effects and exposure to PCBs. The reports he discussed dealt with diverse potential health effects. With respect to liver effects, he concluded that "Alterations of liver function and fat metabolism associated with PCB exposure have been observed in several studies, but are characterized by investigators as mild and of no clinical significance." He also concluded that "Mortality studies concerned primarily with cancer present problems of interpretation due to the small sample size of some of the studies, and to the confounding effect of other exposures. However, they do exhibit a pattern, which is that none of the studies agree on the cancer sites at which an excess mortality was found, and the excesses that ` 34 " SCM 019686 OHO^'O WATER PCB-SD0000012957 were found are in general not statistically significant. One must conclude that the findings of the mortality studies reflect a sporadic pattern of excess mortality at different sites which is not consistent with a carcinogenic effect of PCBs. In addition, where an examination of duration and latency of exposure was possible, no association with these variables was found [32]7 . ' "Taken as a whole, the epidemiologic studies find that high occupational exposures to PCBs may cause dermatitis of various kinds, but that there are no other clinically observable effects, including the occurrence of cancer." Summary Some remarks by Cole and Merletti (1980), although written in a more ` general vein, appear to be particularly suited for ending a discussion on the potential carcinogenicicy of PCBs. "In view of the difficulty of deciding whether a "suspect" carcinogen is in fact a weak carcinogen or in fact harmless, we point out one aspect of this scientific judgement which, though well known, is often lost sight of; namely, that in prin ciple, it is more likely that a non-carcinogen will appear to be a weak carcinogen than the reverse. This asymmetry should be fully appreciated by legislators and regulators. It is summed-up succinctly, if not very accurately, in the oft-heard phrase "you can prove a positive but not a negative". This is clearly illustrated in the case of PCBs. Review of the results of a large number and wide range of chronic feeding and metabolism studies in animals and of mutagenicity studies in various systems has failed to establish that PCBs are carcinogenic. However, - 35 - SCM 019687 (JHM.iYi WATER PCB-SD0000012958 presently available rodent data raise some suspicions and lead to disputes about the carcinogenicity of PCBs. Attempts to compare and evaluate results of rodent studies reported in the literature are complicated by different uses of similar terminology and variations in the criteria employed for diagnosing hepatic tumors in rodents. Co-carcinogenesis studies have reported both promotion and inhibition of tumors in rodent livers following prolonged administration of PCBs in conjunction with an initiator. Thus, it is not likely that animal studies and other laboratory procedures will lead to a universal consensus on the carci nogenicity or non-carcinogenicity of PCBs. While epidemiology studies of humans exposed to PCBs present problems of interpretation due to their small sample size and the confounding effect of other exposures, they present the best available evidence for assessing the carcinogenic potential of PCBs to humans. Epidemiology studies, including recent studies involving large cohorts with lengthy exposures to PCBs, demonstrate a pattern that is not consistent with a carcinogenic effect of PCBs. - 36 - SCM 019688 WATER PCB-SD0000012959 Footnotes 1. Farber, E. (1973). Hyperplastic liver nodules. In:Methods in Cancer Research, Vol. 7, H. Busch, ed. Academic Press, N.Y. pp. 345-375. 2. See reference: Squire, R.A. and Levitt, M.H. (1975). 3. See reference: NCI (1978). 4. Grube, D.D., Peraino, C., and Fry, R.J.M. (1975): The effects of dietary phenobarbital on the induction of skin rumors in hairless mice with 7,12- dimethylbenz(a)anthracene. J. Invest. Dermatol., 64. 258-262. 5. Costa, M., Costa, E.R., Manen, C.A., Sipes, I.G., and Russell, D.H. (1976): Adenosine cyclic 3',5'-monophosphate-dependent protein kinase and ornithine decarboxylase involvement in the induction of cytochrome P-450 and hepatic hypertrophy. Mol. Pharmacol., 12, 871-878. 6. Delor is a tradename for PC3s made by Chemko in Czechoslavakia. 7. Brown, D.P. and Jones, M. (1981). Mortality and industrial hygiene study of workers exposed to polychlorinated biphenyls. Arch. Environ. Health 36, 120-129. SCM 019689 000373 WATER PCB-SD0000012960 References L. Anon. (L98L). The Toxicology of PCB's. An overview wich emphasis on human health effects and occupational exposures. Hazard Evaluation System. Epidemiological Studies Section. State of California. Berkeley. 2. Bahn. A.K., Grover, P., Rosenwaike, I., O'Leary, K., Stellman, J. ' (1977) PC3? and melanoma. Hew Engl. J. Med. 296, 108. 3. Bahn, A.K., Rosenwaike. I., Herrmann, H., Grover, P., Stellman, J. and O'Leary, K. (1976). Melanoma after exposure to PCB's. Hew Engl. J. Med. 295, 450. 4. Berry, D.L., DiGiovanni, J., Juchau, M.R., Bracken, W.M., Gleason, G.L., and Slaga, T.J. (1978). Lack of tumor-promoting ability of certain environmental chemicals in a two-step mouse skin tumorigenesis assay. Res. Commun. Chem. Pathol. Pharmacol. 20, 101-108. 5. Berry, D.L. Slaga, T.J., DiGiovanni, J., and Juchau, M.R. (1979). Studies with chlorinated dibenzo-p-dioxins, polybrominated biphenyls and polychlorinated biphenyls in a two-stage system of mouse skin tumorigenesis: potent anti-carcinogenic effects. Ann. N.Y. Acad. Sci. 320, 405-414. 6. Blazak, W.F. and Marcun, J.B. (1975). Attempts to introduce chromosomal breakage in chicken embryos with Aroclor 1242. Poultry Sci. 54, 310-312. 7. Brinkman, V.A. Th. and deKok, A. (1980). Production, properties and usage. In: Halogenated biphenyls, terphenyls, naphthalenes, dibenzodioxins and related products. R.D. Kimbrough, ed. Elsevier, Horth-Holland, Hew York. pp. 1-40. 8. Calandra, J.C. (1976). Summary of toxicological studies on commercial PCB's. National Conference on Polychlorinated Biphenyls, November 19-21, 1975. Chicago, Illinois. EPA Report No. 560/6-75-004. March. NTIS PB-253 248. pp. 35-42. 9. Cole, P. and Merletti, F. (1980). Chemical Agents and Occupational Cancer. J. Environ. Pathol. Toxicol. 3, 399-417. 10. Danz, M. and Urban, H. (1980). Elevated mitotic number in the adrenal certex as a reflection of early events in growth induction by carcinogens and promoters - a possible short-term assay. Further studies in the assessment of toxic actions. Arch. Toxicol. Suppl. 4, 19-21. 11. DHHS. Public Health Service (1980). Health Effects of Toxic Pollution: A report from the Surgeon General. Serial No. 96-15. August. U.S. Government Printing Office. SCM 019690 000374 WATER PCB-SD0000012961 12. DiGiovanni, J. , Viaje, A., Berry, D.L., Slaga, T.J. , and Juchau. M.R. ( 1977). Tumor-initiating1 ability of 2,3,7,8-tetrachIorodibenzop-dioxin (TCDD) and Aroclor 1254 in the two-stage system of mouse skin carcinogenesis. Bull. Environ. Contain. Toxicol., H3, 552-557. 13. Dikshith. T.S.S., Rockwood, W., Abraham, R., Coulston, F. (1975). Effects of a pplychlorinated biphenyl (Aroclor 1254) on rat testis. Exp. Mol. Pathol. 22, 376-385. 14. Gaffey, W.R. (1981). The epidemiology of PCBs. Presented at the Annual Meeting of the American Chemical Society. 15. Garthoff, L.H. ,'Friedman, L., Farber, T.M., Locke, K.K., Sobotka, T.J., Green, S., Hurley, N.E., Peters, E.L., Story, G.E., Moreland, F.M., Graham, C.H., Keys, J.E., Taylor, M.J. , Scalera, J.V. , Rothlein, J.E., Marks, E.M., Cerra, F.E., Rodi, S.B., Sporn. E.M. (1977). Biochemical and cytogenetic effects in rats caused by short-term ingestion of Aroclor 1254 or Firemaster BP6. J. Toxicol. Environ. Health 3, 769-796. 16. Goldstein, J.A. (1980). Structure-activity relationships for the biochemical effects and the relationship to toxicity. In: Halogenated biphenyls, terphenyls, naphthalenes, dibenzodioxins and related pro ducts. R.D. Kimbrough, ed. Elsevier/North-Holland New York. pp. 151-190. 17. Green, S., Carr, J.V., Palmer, K.A. and Oswald, E.J. (1975a). Lack of cytogenetic effects in bone marrow and spermatogonial cells in rats treated with polychlorinated biphenyls (Aroclors 1242 and 1254). Bull. Environ. Contain. Toxicol. 13, 14-22. 18. Green, S., Palmer, K.A. and Oswald, E.J. (1973). Cytogenetic effects of the polychlorinated biphenyls (Aroclor 1242) on rat bone marrow and spermatogonial cells. Toxicol. Appl. Pharmacol. 25, 482. 19. Green, S., Sauro, F.M. and Friedman, L. (1975b). Lack of dominant lethality in rats treated with polychlorinated biphenyls (Aroclors 1242 and 1254). Food Cosmet. Toxicol. L3, 507-510. 20. Hansell, M.M., Ecobichon, D.J., Comeau, A.M. and Cameron, P.H. (1977). The relationship between retention of pure chlorobiphenyl cogeners and hepatic function in the rat. Exp. Mol. Pathol. 26, 75-84. ' 21. Hargraves, W.A. and Allen, J.R. (1979). The in vitro binding of 2,2',5,5'-etrachlorobiphenyl metabolites to rat liver microsomal pro teins. Res. Comm. Chem. Pathol. Pharmacol. 25, 33-52. 22. Heddle, J.A. and Bruce, W.R. (1977). Comparison of tests for mutagenicity or carcinogenicity using assays for sperm abnormalities, formation of micronuclei, and mutations in Salmonella. In: Origins of Human Cancer. Cold Spring Harbor Conferences on Cell Proliferation. H.H. Hiatt, J.D. Watson and J.A. Winsten, eds. Cold Spring Harbor Laboratory. Vol. 4 Book C, pp. 1549-1557. - 39 - SCM 019691 WATER PCB-SD0000012962 23. Hendricks. J.D., Putnam. T.P.. Bills, D.B., and Sinnhuber. R.O. (1977) Inhibitory effect of a polychlorinated biphenyl (Aroclor 125-1) on Aflatoxin 31 carcinogenesis in rainbow trout (Saimo gairdneri). J. Nad. Cancer Inst. 59, 1545-1551. 24. 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(1973b). Histopathologic studies on liver tumorigenesis induced in mice by technical polychlorinated biphenyls and its promoting effect on liver tumors induced by benzene hexachloride. J. Natl. Cancer Inst. 5J., 1637-1646. 29. Ito, N., Nagasaki, H., Makiura. S. and Arai, M. (1974). Histopathological studies on liver tumorigenesis in rats treated with poly chlorinated biphenyls. Gann 65. 545-549. 30. Ito, N., Tatematsu, M., Hirsoe, M. Nakanishi, K. and Murasaki, G. (1978). Enchancing effects of chemicals on production of hyperplastic liver nodules induced by N-2-fluorenylacetamide in hepatectomized rats. Gann 69, 143-144. 31. Keplinger, M.L. , Fancher, O.E., Calandra J.C. et al. (1972). Toxicological studies with polychlorinated biphenyls. Read at PCB conference, Quail Roost Conference Center, Rougemont, North Carolina, 20-21 December 1971. Cited in Polychlorinated Biphenyls Environmental Impact. A Review by the Panel on Hazardous Trace Substances. March, 1972. Environ. 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National Research council of Canada, Publication no. NRCC 16077. 74. Safe, S. (1980). Metabolism, uptake, storage and bioaccumulation. In: Halogenated biphenyls, terphenyls, naphthalenes, dibenzodioxins and related products. R.D. Kimbrough, ed., Elsevier/North-Holland Mew York. pp. 81-107. 75. Squire, R. A. and Levitt, M.H. (1975). Report of a workshop on classification of specific hepatocellular lesions in rats. Cancer Res. 35, 3214-3223. ' 76. Stadnicki, S., Lin, F.S.D. and Allen, J.R. (1979). DMA single strand breaks caused by 2,2',5',5-tetrachlorobiphenyl and its metabolites. Res. Comm. Chem. Pathol. Pharmacol. 24, 313-327. 77. Stott, W.T. and Sinnhuber, RiO. (1978). Trout hepatic enzyme activation of aflatoxin B x in a mutagen assay system and the inhibitory effects of PCB. Bull. Environ. Contain. Toxicol. 19, 35-41. 78. Sugimura, T., Kawachi, T., Matsushima, T., Nagao, M., Sato, S. and Yahagi, T. (1977). A critical review of submammalian systems for mutagen detection. In: Progress in genetic toxicology. D. Scott, A.B. Bridges, and F.H. Sobels, eds. Elsevier/North-Holland Biomedical Press, pp. 125-140. 79. Sugimura, T., Sato, S., Nagao, M., Yahagi, T., Matsushima, T., Seino, Y., Takeuchi, M., and Kawachi, T. (1976). Overlapping of carcinogens and mutagens. In: Fundamentals in Cancer Prevention. P.N. Magee, S. Takayama, T. Sugimura and T. Matsushima, eds. Univ. of Tokyo Press, Tokyo/Univ. Park Press, Baltimore, pp. 191-215. 80. Tazima, Y. (1980). Chemical mutagenesis in the silkworm. In: Chemical Mutagens: Principles and Methods for their Detection. Vol. 6, F.J. deSerres and A. Hollaender, eds. Plenum Press, N.Y. pp. 203-233. 81. Turusov, V.S. and Takayama, S. (1979). Tumours of the liver. In: Pathology of Tumours in Laboratory Animals. International Agency for Research on Cancer. World Health Organization. Lyon France. V.S. Turusov, Editor-in-Chief. Vol. II-Tumours of the mouse, pp. 193-233. 82. Uchiyama, M. and Chiba, T. (1974). Co-carcinogenic effect of DDT and PCB feedings on methylcholanthrene-induced chemical carcinogenesis. Bull. Environ. Contain. Toxicol. 12, 687-693. - 44 - SCM 019696 000380 WATER PCB-SD0000012967 83. Wasserman, D., Miller, H.J., and Wasserman, M. (1978). . Polychlorinated biphenyl-induced rat liver adenomas. Toxicol. Eur. Res. 1. 159-172. 84. Weisburger, J.H. and Williams, G.M. (1980). Chemical Carcinogens. In: Casarett and Doull's Toxicology, J. Doull, C.D. Klaasen, and M.O. Amdur, eds., Macmillan Publishing Co., Inc., New York, pp. 84-138. 85. Weltman, R.H. and Norback, D.H. (1979). Hepatic ultrastructural changes of rats exposed to polychlorinated biphenyl isomers: Hexachlorobiphenyl-induced neoplastic nodules. Toxicol. Appl. Pharmacol. 48, A181. 86. Wong, A., Basrur, P.K. and Safe, S. (1979). The metabolically mediated DNA damage and subsequent repair by 4-chlorobiphenyl in Chinese hamster ovary cells. Res. Commun. Chem. Pathol. Pharmacol. 24, 543-550. 87. Wyndham, C. Devenish, J. , Safe, S. (1976). The in vitro metabolism, macromolecular binding and bacterial mutagenicity of 4-chlorobiphenyl, a model PCB substrate. Res. Comm. Chem. Pathol. Pharmacol. 15, 563-570. 88. Yoshimura, H., Yoshihara, S., Ozawa, N. and Miki, M. (1979). Possible correlation between induction modes of hepatic enzymes by PCBs and their toxicity in rats. Ann. N.Y. Acad. Sci. 320, 179-192. SCM 019697 0^0 rri WATER PCB-SD0000012968 TABLE 1 PCBs: Tal>ulalion of Liver Nodules Reported in Mice1 Approx, wt. % Cl2 Product Strain Dosing Duration PCB in diet (ppm) No. Animals 52-54 Aroclor 1254 BALB/cJ 6 mos3 11 mos 0 300 0 300 34 24 24 22 Kanechlor 500 dd 32 weeks 0 100 250 500 6 12 12 12 Kanechlor 500 dd 32 weeks 0 100 250 500 20 18 20 17 Kanechlor 500 dd7 32 weeks 0 100 250 500 12 19 20 17 Nodular Hyperplasia - 0 0 0 7 9 4 Hepatoma 0 1 0 9< (-)5 '- - Hepato cellular Carcinoma - 0 0 0 5s 0 0 0 76 0 0 0 0 Reference Kimbrough +,Linder (1974) Nagasaki, et a 1. (1972) 1 to, et al. (1973a.b) Nagasaki, et a 1. (1974, 1975) Nagasaki, et a 1. (1974, 1975) 48 Kanechlor 400 dd 32 weeks 0 6 100 12 250 12 500 12 0 0 0 0 - Kanechlor 400 dd 32 weeks 0 100 250 500 0 17 19 20 -- - . - Kanechlor 400 dd7 32 weeks 0 100 250 500 12 20 20 17 _ `- - - - 0 Nagasaki, el al. 0 (1972) ' 0 Ito, et_aK (1973a,b) 0 0 Nugasaki , cljL 0 (1974, 1975 S ' 0 0 0 Nagasaki , i-L _a I . 0 (1974, 197*)) " 0 0 SCH 0 1 9 6 9 8 WATER PCB-SD0000012969 TABLE 1-continued Approx. wt. X Cl2 Product Strain PCBs : Tabulation of Liver Nodules Reported in Hice1 Dosing Duration PCD in diet (ppm) No. Auima1s Nodular Hyperplasia Hepatoma llepatocellular Carcinoma i Refereure 40-42 Kanechlor 300 dd 32 weeks 0 100 2.SO 500 6 12 12 12 o' 0 0 0 . 0 Nagasaki, e i a 1 . - 0 (19/2) - 0 [to, eL al . ( 1973a,h) *0 t Kanechlor 300 dd 32 weeks 0 100 250 500 20 19 19 20 - - - - 0 Nagasaki, eL a 1 . - 0 (1974, 1975) -0 "0 Kanechlor 300 dd7 32 weeks 0 100 250 5110 12 19 20 20 - - - 0 Nagasaki, el a 1 . - 0 (1974, 1975) -0 *0 1 Hales, except as noted. 2 From Brinkman and deKok (1980) * Held an additional 5 months before sacrifice. 4 Ho. of animals. One had 2 hepatomas for a tumor total of 10. 8 Apparently same data reported as hepatoma and hepatocellular carcinoma. See references and text. a Apparently same idata reported as nodular hyperplasia and hepacellular carcinoma or Lumor. See references ami text. 7 Females. IA o z o o o Q -- *0 0" 00 CJ WATER PCB-SD0000012970 ! FCIIs; T*i|uI d i i ml of I.iver Nmlulcu Hl`|>orted in HuLs Approx. wt. X Cl1 Product. PCB Dating in <liel No. Arii'iiouidloua Nodular Strain Sen Dural ion _!ppiu)___ Aiiiiu 1 s Nodule* llyjicrj11 us i a 60 Aroclur 1260 SlicrsMit V 21 BOS.^ 0 in 100 184 - _ - Aroclor (21*0 Churle* M, 24 uos. Hi vcr K 0 1 10 100 21 26 25 25 - I 0 7 6 212,.414,15,5` - _ HdvacbloroNi phciiy la - 24 suit. 100 - - _ Ni*op 1 dbl i c Nodu 1 tts Ad*ilouii Ilepul Iiiuj llepaioic 1 1 u 1 j r 0d 1 11 lliMU.I Clio- 1 lli-p.ll HUd Hr I uviu r 0 __ 1 _ K i win ougit, 144 " - 26 - _ _0_ - -0- - 1- -7- < _ 0 l.:v mukd* * 0 (l`J8l) (1 4 . We I LuiiiM 6 Nuriidtk (|j/j) 62-64 Aroclor 1264 Chorion N. 24 nos. 0 2'J 1 0 0 I.eviitKkdte Ktver F 1 30 - 0 ' - - 0 - 0 (luai) 10 26 - 3 - -0. 0 100 26 - 14 * -4- 2 Aroclor 1254 11 ocher N 105 weak* 0 24 . . .0 0 m NCI (19/81 144 26 24 - - - 0-0 . SO 24 - - - 0 - 1 _ 100 24 " - " 1-2 - Aroclor 1254 Fischer F 105 week* 0 21 - _ 0__ 340 26 24 - - - 0-- NCI (I'J/h) so 22 - - - 1-. _ 100 24 " - " 2- - - Aroclor 1264 l.ucti 1 F 28 uui.6 0 u . _ _ 0_ _ W.i | tU.ll , 200 6 - - " - - - 4-i .l. ( ri/n) Kuitcclilur UiiiUr n 28-52 wk 0 )H 0 _ _._ I I M . 4* I I 600 Ion 2* - 1 - -- - ( n/O Mil) )< - 6 - --- . 1000 11 6 - --- _ WATER PCB-SD0000012971 SCM 0 1 9 7 0 0 I TAIII.K 2-cunl iiiU4*I PCjjs: Taintjat ion u| Liver Nullities Kejioriril iii Hal a A||iroM. wi_. 1 Cl ProUmt______ ___ Si rain PCll Dosing in ill el Nu. Ailriiiiui.il mis Noiiu 1 a r Sex Dural ioii _!pi*") . An im.i l * Nmln 1 rs ljy|ii`i|> l .ik 1 a 48 Kaiicclilor Donryu ti 23-62 wks 38.5- S 0 - 400 616 10 0 . - K 50 - 10 6 - Kaiiekiilor Wtklar ti 211-52 vkx U 100 500 I0U0 IH 16 8 10 . - 0 2 0 3 Ni*i*|i | a si ii: Nmliili'k Ailriiuma llrjiat niuj Ilrji.i lOi'i* 1 Id 1.11' Ca 11 1 unui.1 CIlO|.ii4KmJ| 1' |4. 1 Wtll.l !<> 1 i* i i'i - - - - - K iiiuii a - - - - - (l*J/ 1) - --- - - --- - . .. _ l 1 it , r - - - - - ( I8J4> - --- - - --- - 40*42 Aroclor 1242 (Mur lex M. 24 aio*. lliver K 1 10 100 Kanechlor Vixlar H 28-52 wka 0 100 500 1000 2,2'.S.S'-lelra- chlorobi|*lieiiyl* - 24 aiox. 100 23 22 28 18 18 22 1!) IS - - - - - 1 1 1 B 0 1 0 0 - - K - 0 I.I*V 1 tlh kata - 0 - (1 ( I8HI) - - 0 - II - * 3- - 1 - _ - _ _ llu, 1-1 i 1 . - - - - - 118/4)' - --- - - "-- - - - - _ - Wi: 11 man 6 NuiLaik ( l`W' 1 Kroai briukmau and JeKuk ()!)80). * llrlil an adi111unal 2 uutiilii* liclurc sat*ri I ii`:. 3 Absirail. Drlaili. IiuhIciI. * I'lckrnic rcj<orlcii. * Llbanul kuiuiiuu in i|i Hiking wjUf. SCM 019701 WATER PCB-SD0000012972 SCM 019702 Product Jfaneclilor 400 Kaneclilor SOU Kdiittlilor SOU Kaneclilor 400 Kaneclilor S00 Kauechlor S00 Aroclor I2S4 Kaneclilor S002 Aroclor I2S4 Kaneclilor S00 Kunechlur SOU Kuiurt.li lor SOU Kuiict li 1 u r SUO Mode Diet Diet Diet Diet Diet G.v.ge Diet Guvage Dicl DitrL Uid Did Hid Tme Willi WlLll Willi Willi Willi After After lie l ore Willi Willi Willi lie-1 urc A 1 l i Treatment1 Mode 2a-MeCll if-line Hi rriiii: 1111(11 Util Hid |i-unc Hint o-lillC OieL UEN Wilier DEN W.ier HEN Water DEN* Water AFU, II id 2-KAA Did J'-Me-IIAII Dili llii'i r-Mc-HAII Uni tabeej co-cakcinodkhe.sis studies urm mu Specie* Observalious \i[cl . dll aiou*e tlt| mouse tld woutfO dd mouse SpragueHawley rat Wislar rat SpragueHawley ral Winter rat . Ha lidiuw l root Sju agucHaw 1 try l`al S|*| aKiu. ll.twlty rat S|i I ilgut: " li.iwlry iat Spi ........ Il.tw l ry ill lli'jijttt*i*11 ii 1 ar carcinoma witli 20~hcCb. Nolle Willi combination. Combination im ri*anrtl lu*jal of* 1 1 u 1 ar i'aiciiitiiua over U-llliC ulniur. (luuliiiiiil lifii priubiced Hepatocellular Ca reiiioiua . Each alone negative. Combi iial ion increased liepalucel inlar carcinoma over u-UIIC alone* Hepatocellular carcinoma with DEN. None with combination. Combination increased be|ialocel lular carcinoma over HEN uloue. Combination increased |i<?putoci;l Inlar carcinoma over HEN alone. Combination decreased liepatorcl lular carcinoma m o(lti|iring. Ilcpul oi-<:l lular cjrcinoma with AKMj. None with combination. Hcpalorcl lular carcinoma with 2-KAA. None with Combinalion. Hepatocellular t .in i iioiua witli J1-flc-HAII. `Nniie with Couili 1 Hal i on . Nepal ii i* 1 lular t`a i*i` I iiouia wi 1 It 1* -H*~liAN. Nuni! witli comb l leal i on . i'tiiiib i ii.ii mu | m*i t*.|.M*d ln-|(.i in i*| 1 tt 1 >m i a I > i ii"i.i uvi'i 1 * - ft** - t lAtl alom*. 111 Ii 1 yama , and Club.*, l`J/4 11 ti , cl a 1 . rjn.,1. Nagas.ik 1 , l .il 1 l`J/A, !>/". 1 1 n i 1*1 a 1 Nagatfaki, tl ;j. I*JK, 1 J/*> Nagasaki, et dj. l`J/4, lJ/S flnkiura, el ul: |*)?4 . Ninhittuini, id76, i. Prtrdou, el a j. I*)H| Ninhiauiui, )`Jtt0 llendr i ck*, et al. l'J/7 Hukiura, t* 1 nl. l`J/4 Mak l in a , t* 1 -I. l'l/4 - K imm a , cl .. I. pj/o Kimihi ,i, d .. 1 pi/n K 1IIIO 1 .t ( | ( ..1 !'/>. WATER PCB-SD0000012973 H 019703 000.<S v Product Kaneclilor 500 Kaneclilor 500 Kanechlor 500 Kaneclilor 500 Kenechlor 500 Unspecified Kaiiethior Aroclor 1254 Aroclor 1254 Aroclor 1254 Ardor 1254 Ai or 1 mi 1254 A.oi |r 12^4 Hode Tine Other Treatment 1 TAMI.K d-cuiil iuucd SMHHAKY OK CQ-CAIllllNOGK.NKSIS SHIM IKS WITH Mis Mu.le Species Observalions He 1 . Diet With Old With Cavage Cevege Cavage out After l> Vi Ik After k With After k With Alter k With Skin Skin Sk in Alter lie fore before Mid Mid Mid ltd ore lie lure lie I *t e V-Me-UAU * UKN Mid Sprague* Hawley rat 2-KAA t 0KN IlkCL Sprague** Hawley rat MKN Voter WisLar MOT Cavage ral MKN Mi'll 0KN Mi'll, MOT Water Water Water Water, liovuge Wislur rat Uislur ral 2-FAA Diet ParLiai licpalecloaty Ki seller ral 7,12-OMBA Skin CHI Mouse TPA .Skin C|)l mouse 7,12-MhMA i ri'A Skin CHI mouse Walker 256 llll fd* tumor ir| Ik uiuticu lor hSU a:l U 1 I mlr-" ifirui la r hit | |rttki'iU|.| Vlllle Ink i a|** 1 t 1 WHIM 1 Sp> agneHaW 1 ry i.il i:miii./i. hm hi :.t: ll.illt/i Cutultinal iuii leereasird lu*pa(urr 1 1 u 1 ar carcinoma over ii'-Hc-DAli * Hl.N. Cumbkiiat tun decreased lurpal ore 11 u la r caicinema over 2-KAA i UKN. Combinalion increased hrpaloccllular carcinoma over HKN * UHT. Cnuiltlnat ion increaseil hepatocellular carcinoma over HCN SHU. CombiusLion increased Impatorellular carcinoma uver UKH, SPB, HHT. harked Increase In ltyper|laslic liver itudu lus. Mukiuru, cl .. 1. l!)/4 flak i lira, el al. I*J?4 * Ni kli i fciUHl , I974ja,b Hislti ttiinii , l4J /'Ja Ji Nishiaimn, l')7`Ju , b llu, dM. 1>J7H Not a promoter of skin iuittors. Utile or no skin Inutor Jiti t iat ion. Nu di 1 lereitce or derreaae in skin Luutor Uinductiuu over },I2~HHIIA TI'A, dependiitg on prel lealuieiil inleival. Tnuiur growl It inhibited. `Survival lime increased. 1 Kubant'c.l giuwtli ul MSII iianr. Dili 4411 .(llrii um ogrtii*:. i u nl hxlimry 1 enki'ni t a v 11ns. Merry, d ul. I'J7U, l`J7`J Ilf r iy t i:l al. |4J7'J Did ovaiiii i . i*l al. 1`JJ? llerry, el al. I4JJ'I DiCiuvaiuii, el al. |4)77 Kei kv 1 tel ami K liutiir lilui t , 1 *1 / / .4 , |i Knrkviiel ami K.tlln , |'J/fl K..II.-I, IS// WATER PCB-SD0000012974 f Product Aroclor 1242 Aroclor 1221 Mode Tine TAljU_3-co.il limed SUHHAHY OK CO-CAHCINOCKNI-SIS STUH1ES WITH Pflis Other Treatment 1 Mode Species Ohscrvalions hut Utel lit: 1 ore llclore Moloney 1 eokeiui a virus Holooey leukemia vi rus lot Ca pe i i L micj 1 lla Ih/C mouse 1 lit l a pe* ilooea1 Italh/c mouse Hid not allim'1 oncogenesis ol Moloney leukemia vuus. Hid not ailed oncogenesis of Moloney leukemia virus. 'Chemical* used: 20-MeCh s 20-mclhylcliolanihrene a-BIlC a a-benzena hesachlorIda p-BtlC * p*benten licMschloride l)N dielhylnitrossmUc AFB| * aflUuxin Bt 2-FAA * N-2-fluorcnylacclamidc 3*-Me-DAB = 3'-aeiliy l*4*diaelliylamlno*gobeiucne DOT = dichlorodiphenyllrichluroelhaiie SPB = sodium phenobarbilal 7,12-DHA - 7,2-d(metliy Iht?iu|<i|anthracene TI'A - 12-O-lel radecanuy Iphoi l>o) - I'J-acelat e ^Pregnant dams dosed with Kaucihlor 4H0. Offspring dosed with DCN. ^Established tissue culture cell line derived from Moloney sarcoma vi rus-induced tumors in H6 mice. He I . Fuller, 1 *1 / / Ko1 let, l'J/7 SCH 0 1 9 7 0 4 WATER PCB-SD0000012975 i Tester Strain Product AKDCI.OK 1268 TABLE 4 PCIls :_TalinI*I ioii_ol'__Mieroliia j__Mulageiiicily Tcsls C3076 D3052 S. (yjdi i sin r i ow G46 TAOS TA100 '"TAtOUU TAI535 TAIS36 IA1537" TAI5JB Neg. E. roll UF2 WP2 uvrA- Hcl . Uyndliaiii, el a 1 . t*1 /o AKOCI.UN 1254 AHOCLOK 1254 AH0CI.UK 1254 Kaneehlor 500* K.iieclilor 500 Neg. Ncg. Neg. Neg. Neg. Neg. Neg. Neg. Neg. Neg. Neg. Ncg. Neg. Neg. Neg. Neg. Neg. Pox* Neg. Neg Neg o Neg. Pi ulsl , cl at. 1 *1U | llctidlc and IlfuCe \lH / Wyndliaui, el_al . |4 7t Odasliiuia, 1476 Sugiiuura, cl al. I'J/o 2,2*.5,5-tetraclilorobiphenyl Neg. Wyndliaui, cl d|; |47n 2,2* 5,51-let raclilorobiplicny 1 Neg' Neg* Neg1 Neg' K.ncchlor 300* Neg. Neg. Neg. Neg. K.nechlor 300 Neg. Neg. AHOCLOK 1221 . Hum* 4*chlorobiphenyl * Ho#' !f I! J1" 15 SCM 0 1 9 7 0 5 4-chlorobipheuy1 He*. He*. I. All icsm done with aud without Neg. Neg. Neg. Neg. activdlion except is noted. Neg. Neg. 2. oilier xiraim# icxlcd, xce lexl and relcrcnee. = Nol lex led. Ncg. - Ncgaltve. Ncg - Only leviedwiilioul dilivalioii. l Neg' s Only Icsled withai l I val i nil. I'uit1 - |`os ill ve only with .*l i v.il i mi. ` lliit i* i I .i 1 It. 'i*:w lexl. t Ncg o Neg. Neg. Hail., rl i|. I'JIU Oilaxluuia, 147b Sugliunra, el al. |47 Uymlliaiu, el al. 147b Wyudliuui, cl al. 1471* ttittalnui, :l al. 14 74. WATER PCB-SD0000012976 ILU X I I i <! WATER PCB-SD0000012977 decision Analysis. Phil 's -eroy, A. A.. Gaylord, rger, T. 1982. Reducing nduced hospilalizatioa: utilization review. Drug 986. Pharmaceutical inenehc dnig competition icon of the Drag Price id Patent Term Restota14. Pharm. Med. 1:177- :tolIcy, P. D., Brown, T. usubstitution law conutioo? Ann. Intern. Med. 1986. Med. Lea. 28:1-2 Ann Rev. Pharmacoi. Toxicol. 1987 27.87-111 HUMAN HEALTH EFFECTS OF POLYCHLORINATED BIPHENYLS (PCBs) AND POLYBROMINATED BIPHENYLS (PBBs)1 Renate D. Kimbrough Center for Environmental Health. Centers for Disease Control, Public Health Service, US Department of Health and Human Services. Atlanta. Georgia 30333 INTRODUCTION Polychlorinated biphenyls (PCBs) are chemical compounds with the empirical formula C|2H|o_,Cln, with n = l--10. They are a mixture of chlorinated biphenyl congeners. Theoretically. 209 such congeners are possible, but at least 20 congeners have never been identified in commercial products. In addition. PCBs may contain polychlorinated dibenzofurans and chlorinated quaterphenyls as impurities. PCBs were discovered before the turn of the century, and the useful industrial properties of mixtures obtained by chlorina tion of biphenyl were recognized early. In 1966 the discovery of PCBs in environmental samples (l) spurred renewed interest in the analysis and toxic ity of these compounds. In recent years many industrial nations have taken steps to control the flow of PCBs into the environment. PCBs and PCB-containing formulations are restricted (an exception is sometimes made for mono- and dichloro-PCB) for most uses, except for categories such as closed-system electrical equipment and hydraulic fluids in mining equipment. Commercial production of PCBs began in the United States in the late 1920s. In 1971, Monsanto Chemical Company voluntarily stopped openended uses of PCBs, and subsequently only the lower chlorinated biphenyls 'The US Government has the right to retain a nonexclusive, royalty-free license in ind to any copyright covering this paper. 87 2 DEPOSITION * EXHIBIT I 1 S iD/( ooorro WATER PCB-SD0000012978 88 KIMBROUGH HUMAN HEALTH EFFECTS OF PCBs & PBBs 89 were produced (Aroclor 1242 and 1016). In 1977 ihe company ceased produc tion enlirely (2). Many PCHs manufactured in Ihe past (3) are still in use in old transformers, but even this use is decreasing. The estimated cumulative production and consumption of PCBs in Ihe United Slates in the period 1930-1975 (in millions of pounds) was as follows: total production, 1400; imports. 3; domestic sales. 1253; exports, 150. PCBs are inert chemicals that are fairly resistant to degradation. Because of their stability and lipophilicily, they have accumulated in the environment and in organisms. They have been identified in indoor air (4) at concentrations of into feed for livestock. The flame retardant was called Fircmaster and Ihe magnesium oxide. Nulrimaster. In 1973 some bags of Firemaster were accidentally sold as Nutrimaster and mixed into animal feed. This resulted in widespread contamination in the slate of Michigan (14). Since 1974 PBBs have not been produced in the United Stales, At the time of Ihe exposure little was known about Ihe toxic effects of PBBs. Ten years after the Michigan residents were exposed, no clinical illness has been causally linked to PBB exposure in this group, although chloracne was apparently noted in some workers who manufactured PBB. 0.1 /ig/m1 (5). in fish (6, 7) and other food products, and in sediments from lakes and rivers (8. 9). PCBs have also been identified at varying con HUMAN EXPOSURE centrations in soil (0.01 mg/kg-100 mg/kg) (10). They do not occur naturally. Thus, their presence in Ihe environment is linked with human activities, and Because PCBs are ubiquitous and very persistent in Ihe environment, humans concentrations arc higher in urban and heavily industrialized areas than in have been and will continue to be exposed to them, particularly in in rural and remote areas. However, trace amounts arc also found in remote I dustrialized countries. PCBs may be inhaled in small amounts through ihe air areas, since Ihe air may transport such chemicals over large distances. I or ingested through food. In Ihe United Stales today, people arc primarily Although PCBs are no longer used commercially in the United Stales exposed to PCBs by consuming fish from contaminated waters (9). In the because of their persistence, they are still present in our environment. A past, some farm families were exposed to PCBs from dairy products; these number of transformers and capacitors that contain PCBs. however, are still PCHs originated from coating material used in the inside of silos (15). In in use. Results of laboratory experiments showed that pyrolysis of PCBs at addition, workers who repair transformers and workers who handle toxic temperatures of 200-600C could result in the formation of significant wastes may also be exposed (16). amounts of the more toxic polychlorinated dibenzofurans (PCDFs) (II). The PCB products that were manufactured by Monsanto in the United In February 1981, an electrical fire occurred in a New York State office States had the trade name "Aroclor." The particular kind of Aroclor is building in Binghamton. N.Y. The fire, which originated in a switch gear in identified by a four-digit number. The first two digits refer to the 12 carbon the basement, caused the bushings to crack on a nearby transformer. About atoms, and the second two refer to the percent, by weight, of chlorine in the 180 gallons of PCB dielectric fluid Pyralon (65% Aroclor 1254, 35% chlorin mixture. Thus. Aroclor 1254 contains about 54% chlorine, and Aroclor 1260, ated benzenes and trace additives) were lost. A fine layer of oily soot covered about 60% chlorine. many of Ihe internal surfaces of the 18 floors of the building. Analysis of a The composition of this mixture of chemicals, with different properties, soot sample showed that it contained various isomers of chlorinated di changes once it gels into the environment and into organisms. Some com benzofurans, 2.3,7.8-teirachlorodibenzodioxin, other chlorinated di- ponents of the mixture are more easily degraded in the environment than benzodioxins, and chlorinated biphenylenes. Some of these chemicals are i others. As a result the PCBs identified in Ihe environment resemble Aroclor much more toxic than Ihe PCBs. Because of these findings Ihe Binghamton I 1254 but are not identical to it. Similarly, the PCB mixtures found in humans stale office building was closed, and workers wearing respirators and pro ' usually resemble Aroclor 1254 if exposure occurred primarily through the tective clothing began an extensive cleanup of Ihe building. The cleanup I environment. A different composition of the PCBs may be found in serum or operations lasted four years, and the cost has been enormous (12). Since then I adipose tissue tamplei from occupationally exposed workers. For instance, if several other transformer fires have occurred. These fires have not resulted in the workers arc primarily exposed to Aroclor 1016 or Aroclor 1242, which as much contamination as Ihe one in Binghamton, partly because the contain much less of the more highly chlorinated homologs, then their transformers in Ihe other fires were usually located in a separate.vault (13). gas-chromatographic patterns resemble a combination of Aroclor 1016 or o The problems with polybrominated biphenyls (PBBs). which arc also a o mixture of chemicals, have been quite different. In 1970 a chemical company o in Michigan manufactured polybrominated biphenyls as flame retardants. 'Ihe same company also produced magnesium oxide, a chemical commonly mixed j ' 1242 and Aroclor 1254. For this reason Smith et al (17), in evaluating occupational exposure, divided PCBs into high and low chlorinated biphenyls. The gas-chromatographic pattern of the PCB mixture present in humans can be used to determine whether the exposure occurred primarily o WATER PCB-SD0000012979 . ' .5 ^ 3 3 ) 90 KIMBROUGH through occupation or through the environment, or, in the case of occupation al situations, whether most of the exposure was recent or occurred many years ago. Although 93% of the US population eats fish, the average annual per capita consumption is small: 13 lbs. per year (6. 7). If PCBs are to be quantitated in fish, the edible pnnion rather than the whole Osh must be examined. Because of their lipophilicity, PCBs are preferentially stored in the hepatopancreas of the fish, giving erroneously high levels if the whole fish is analyzed. Sim ilarly, levels in cooked fish are lower (6). Generally, PBBs are not found in the environment because they have had less commercial use than PCBs. PBB contamination is essentially restricted to Michigan's lower peninsula. Most persons who lived in Michigan during the 1973-1974 period have low-level PBB body burdens (18). The greatest degree of contamination occurred mainly in areas with contaminated farms; this segment of the population still has appreciable body burdens (19). Since PBBs and PCBs are lipophilic, they are preferentially stored in adipose tissue. They are also present, to a smaller extent, in serum and other organs and in human milk. The concentration of these materials in different organs depends upon the lipid content of such organs, with the exception of the brain where the concentration is lower than the lipid content would indicate. PCBs and PBBs pass the placenta and ate primarily excreted through bile and milk. In addition to lipid content, the ratios between adipose tissue, blood, and vital organs are influenced by exposure level, sex, age, length of exposure, and also by whether exposure is current. At very low con centrations an analytical imprecision influences the ratios much more than at higher concentrations (19). Since human milk is relatively easy to obtain, it has been used to monitor human exposure. Jensen (20) recently summarized results of such monitoring studies. Average levels of PCBs below 2 ppm (mg/kg) in milk fat have normally been found, although women living in heavily industrialized urban areas may have higher levels. The fat concentra tion in human milk averages 2.6~4.3% (21). At 2% fat, I liter (I) of milk would contain 0.04 mg or 40 fig if the PCBs were present in milk fat at a concentration of I ppm. If an infant weighed 3 kg and imbibed 730 ml of milk per day, it would take in about 6 fig/kg. a dose that exceeds the 1.3 fig/kg dose calculated as acceptable by Cordle et al (6). At 1% milk fat this dose would be reduced to 3 fig/kg. As the infant gains weight, the dose on a kilogram body weight basis will be reduced to some extent, however; milk is the sole food source for only about six months. After the first week, the daily milk intake is estimated to be 150 ml/kg body weight per day. This consumption gradually falls after two months and declines to 120 ml/kg body weight at four-to-six months. Finally, the amount of PCBs and other halogenaied organic chemicals declines with lime. However, al low concentrations this HUMAN HEALTH EFFECTS OF PCBs & PBBs 91 may not be obvious because of continued exposure of the mother and the variability of the analytical results. In addition to PCBs, human milk contains trace amounts of many other persistent chemicals. Whether the infant's consumption of such chemicals has any adverse health effects is not known. Most persons, particularly in industrialized countries, have had some expo sure to polychlorinated biphenyls even if they do not eat fish. The con centrations at which such exposure presents a risk are not clear. Recently, Cordle et al (6, 7) calculated the dose of PCBs to people consuming fish from Lake Michigan. They concluded that persons eating Lake Michigan fish ingested an average of 46.3 mg of PCBs per year; this amount ranged from 14.17 to 114.31 mg/year/person. The calculated mean daily dose received by the exposed group was 1.7 fig/kg/day and ranged from 0.09 to 3.94 fig/kg/ day. Thus, the average spons fisherman consuming contaminated fish would receive a total PCB dose equal to 200 mg in about 4.3 years. No adverse health effects or groups of symptoms clearly related to PCB exposure could be identified in this exposed group. The presence of PCBs in the exposed persons has not caused any observable adverse health effects similar to those observed in the Yusho population (see below). However, this finding does not exclude the possibility that the effects are loo subtle for detection or that they require long-term observation. Similarly, in Michigan an analysis of 1.075 human milk samples showed that all contained PCB residues and that the residues ranged from trace amounts to 5 ppm (mg/kg) based on fat level. The public health significance of PCB residues in human breast milk and their effects on breast-fed infants are unclear. Since there are no human data on which to base public health policy, risk predictions for PCBs have been based on results from animal studies, particularly the positive bioassay studies. Reviewing these data, Cordle et al (6. 7) concluded that a 2-ppm (mg/kg) tolerance for PCB in fish be established, since a I-ppm (mg/kg) tolerance does not greatly reduce the estimated risk. As previously mentioned, some of the isomers of the PCBs and PBBs are much more easily degraded or metabolized. Because they can be metabolized, they are more easily excreted. Others may be retained in the body for long periods; in general, the PBBs appear to be more persistent in human tissues than the PCBs (19. 22). POLYCHLORINATED BIPHENYLS Background When PCBs were first used industrially some workers developed chloracne. Results of early animal studies seemed to suggest that PCBs might have some toxic effects on the liver. Beyond that observation no information was avail- WATER PCB-SD0000012980 o o o CJ fj 92 KIMBROUGH able. Because PCBs were so inert chemically, they were not considered to cause a great deal of toxicity. In 1968 a poisoning outbreak occurred in Japan (23) that affected over 1,000 persons. These individuals had purchased rice oil. in large drums, from a single source and had used this rice oil for cooking. Chloracne was one of the leading signs in those who became ill. It was soon discovered that PCBs had been used as a heat-exchange fluid in the factory where the rice oil originated. PCBs had leaked out of the columns in which they were contained into the rice oil when the rice oil was heated. Since the disease was caused by ingesting contaminated rice oil. it was called Yusho (rice-oil disease). When the outbreak first occurred, its association with exposure to PCBs was not dear. At that time the capabilities for measuring these types of chemicals in tissues and body fluids were limited, particularly in Japan. Therefore, early in the investigation total chlorine, rather than PCBs. was measured. Retrospectively determining the precise dose these patients received is difficult. Whether the consumed oil was uniformly con taminated is also not clear. However, a relationship between the amount of rice oil ingested and some symptoms could be established (24). Because of this poisoning outbreak and other environmental problems, animal studies were started in Japan, in the United Slates, and in other countries to elucidate the toxic effects of PCBs. These data are summarized in several detailed reviews (16. 2S. 26). Animal Studies This article addresses primarily the human health effects of PCBs and PDBs. Therefore we highlight only recent results from animal studies that might give a better understanding of implemented public health policies and potential human health effects. One of the difficulties in using animal data to predict human health effects for PCBs and related compounds is that animal species vary greatly in their responses. Further, many of the animal studies use relatively high doses. Therefore, determining how such animal studies relate to the human situation is difficult. Some animal species, such ax the subhu man primates, the guinea pig, and the mink, are much more sensitive to the toxic effects of PCBs than the rat or the mouse; also the types of toxic effects and morphological changes in the organs of different species vary. Most animal studies conducted during the 1970s used mixtures of PCBs. In general. PCBs were found to affect reproduction and the immune response, and to cause liver tumors in rodents (16). When different mixtures of PCBs were studied, however, the results were inconsistent. For instance, the mix ture Arocior 1234 affects reproduction in rats at much lower doses than does Arocior 1260 (27). More recently some of the isomers of the PCB mixture were found to be much more toxic than others (28-32). The more toxic isomers constitute only HUMAN HEALTH EFFECTS OF PCBs Sc. PBBs 93 a very small portion of the mixture, particularly those with less chlorine by weight, such as Arocior 1242 or Arocior 1016. Recently, Schaeffer et al (33) found that a German PCB mixture--Clophcn A-30, with an average composi tion of 1% monochlorobiphenyl, 20.7% dichlorobiphcnyl, 37.4% Irichlorobiphenyl, 17.3% letrachlorobiphenyl, 1.8% pcntachlorobiphenyl, 1.0% hexachlorobiphcnyl, 0.6% heptachlorpbiphenyl, and 0.1% octachlorobiphenyl--produced a 3% incidence of hepatocellular carcinoma, whereas Clophen A-60 produced a 61% incidence of hepatocellular carcinoma in Wistar rats. The incidence of the disease in the controls was 2%. The Clophen A-60 had an average composition of 0.2% monochlorobiphenyl. 1.1% dichlorobiphenyl, 2.2% trichlorobtphenyl. 3.1% letrachlorobiphenyl. 19.8% pcntachlorobiphenyl, 43.2% hexachlorobiphenyl, 23.3% heptachlorobiphenyl, 4.7% octachlorobiphcnyl, and 0.3% nonachlorobiphenyl. Sim ilarly. Norback & Weltman (34) and Kimbrough et al (33) were able to produce hepatocellular carcinomas in rats with Arocior 1260, the more highly chlorinated Monsanto product. When Arocior 1234 was fed to rats, fewer liver tumors developed in exposed rats (36); however, the incidence of gastric intestinal metaplasia and adenocarcinoma of the stomach increased (37, 38). Whether PCB fractions without hexachlorobiphenyls, heptachlorobiphenyls, and octachlorobiphcnyls produce hepatocellular carcinomas in rodents should be explored. Particularly in the United States, mixtures such as Arocior 1242, 1234, and 1016 were used more than Arocior 1260. Because of these differences in potency, the PCBs in heavily contaminated areas of our environment should be characterized according to their isomeric composition. For instance, whether the PCBs in Lake Michigan are of the same composition as those found in New Bedford Harbor, Massachusetts, is not clear. Aside from tumor formation, PCBs cause a variety of other biological effects, such as the induction of enzymes (39)j In some species they may cause atrophy of the thymus, inlrahcpatic bile duct hyperplasia, hyperplasia of the epithelial lining of the urinary bladder, atrophy of the sebaceous glands, and hyperkeratosis of the ducts (40). Some isomen are feloloxic. and some produce metaplasia of the sebaceous glands, nailbeds, ameloblasts, thymus corpuscles, and gastric mucosa (28, 41). Subhuman primates, mink, and guinea pigs are particularly sensitive to the toxic effects of PCBs; other species, such as the rat, the mouse, and the dog, can tolerate much higher doses. From empirical observations, humans also appear to be less sensitive . to the toxic effects of PCBs. The ability to store these chemicals In adipose t tissue may be protective. Generally, the subhuman primates and mink have ' less adipose tissue than humans. Animals with greater ability to store vitamin A on a quantitative basis, such as the hamster and the rat, are somewhat less susceptible to the toxic effects of these types of compounds (42). The WATER PCB-SD0000012981 o C3 O ID 94 KIMBROUGH mechanism by which these types of chemicals affect hepatic retinoids is not dear. Apparently, the duration of the reduction of hepatic retinoids docs not correlate with (he induced aryl hydrocarbon hydroxylase (Allll) activity (43). Body Burdens Many investigators have reported PCBs in human (issues (44, 43). In the United Stales, according to data from the Centers for Disease Control (CDC), mean PCB serum levels are about 5-7 ng/ml (pbb), although some patients may have higher serum levels without any documented unusual exposure. These data were also summarized by Kieiss (46). Levels in adipose tissue and in human milk fat are 100-200 times as high, since PCBs are highly lipid soluble (20). Mes et al (47) reported that PCB levels in adipose tissue of accident victims ranged from 0.9-9.4 mg/kg. ' Sahl et al (48) surveyed PCB blood levels in 738 pre-employed and 1.058 currently employed workers of a utility company. The median blood level before employment was 4 mg/I and the range. 1-37 mg/I. These levels were quite similar to those in the currently employed group. Patients who died of cancer in Denmark had somewhat higher levels of PCBs in their adipose tissue (49). Since terminal cancer patients have usually lost a great deal of weight, bioconcenlration may have occurred. Similarly, in patients with highly impaired liver function, tissue concentrations of xenobi- olics may be slightly higher than those in healthy persons. However, levels remained higher if parameters such as weight, height, occupation, and resi dence were considered (50), whereas levels of PCBs in breast fat tissue from patients with breast cancer were similar to those of controls (51). Furthermore, Lawton et al (52) demonstrated that random errors and Interlaboratory variations in procedure and methods of data reporting can influence serum and adipose PCB levels. Unless an interlaboratory quality- control system is set up, measured levels between laboratories are not neces sarily comparable. For instance, Lawton et al (52) found that the results of repealed analyses on scrum samples of known composition showed the 95% prediction interval for an individual measurement to be about 42%. This interval depends on the method of extraction, the procedure used, and the means of quantitation. Summary of Human Epidemiology Studies Recently, investigators studied the predominantly black population of Triana. a small rural town in the southern United States (53). This population was excessively exposed to DDT residues by consuming contaminated fish. The residents also had PCB body burdens. Fish consumption correlated positively with PCB blood levels; no other source of PCB exposure could be established. These researchers noted that PCB serum levels increased with age and (hat > , j ! ` HUMAN HEALTH EFFECTS OF PCBs & PBBs 95 levels were lower in females of each age group. Similar findings were made for DDT residues. The serum cholesterol level was positively associated with the log PCB level, independent of age, sex, fish consumption, body-mass index, and alcohol consumption. Rales of borderline and definite hyperten sion for study participants were 30% higher than those expected on (he basis of national rates (54). Log PCB serum values contributed significantly to explaining (he variability of log systolic and diastolic blood pressure in multiple regression analysis (55). Median total cholesterol levels of in dividuals in the United States increase with age from about ISO to 160 mg/di at age 20 to over 200 mg/dl at age 50. PCBs in blood are influenced by serum lipid content, and populations with inherently lower total serum cholesterol levels appear to have a different PCB serum to adipose tissue ratio. The age-associated increase in blood PCB levels could be related to (he long half-life of some PCB isomers that are preferentially retained in mammals (29); as long as exposure continues, a true steady slate between intake and excretion is never reached. Other variables affecting body burdens may be differences in metabolism with age. In the Triana studies, (he blood levels of total DDT residues also increased with age, and others have made similar observations (56. 57). Lawton et al (58) studied workers who had been exposed lo electrical-grade Aroclor 1016, 1242, and/or 1254; the study covered the period from before the workers were exposed lo (wo years after PCB exposure ceased. Serum levels for the lower chlorinated PCBs in 1977 ranged from 57-2270 ppb and in 1979, from 12-392 ppb; for (he higher chlorinated PCBs serum levels ranged from 6-142 ppb in 1977 and from 4-108 ppb in 1979. These findings again illustrate the preferential excretion of lower chlorinated PCB. Lawton et al (58) also found (hat cholesterol levels correlated with log serum PCBs. Similar associations with log scrum PCBs were found for log gamma glutamyl transpeplidase (GGTP) and, in some cases, for log alanine aminotransferase. When the PCB concentrations were expressed as levels in serum lipids, all the associations between serum lipids or enzymes and log serum PCBs disappeared except for those between log GGTP and log PCBs. Similarly, Chase et al (59) found no significant correlation between either serum triglycerides or aminotransferases and the PCB levels in adipose tissue. How age and length of exposure affect these parameters is not adequately explained in the article. Finally, Akagi A Okumura (60) were not able lo confirm a positive association between PCB blood levels and elevated blood pressure in Yusho patients. Thus, as Brown (61) has suggested, the positive association between PCB serum levels and elevated triglycerides and scrum cholesterol can be explained by the increased solubility of PCB in scrum with higher lipid content. In several cross-sectional studies of exposed workers, only minor abnormalities not necessarily related lo PCB exposure have been detected WATER PCB-SD0000012982 cG ^O O O 96 KIMBROUGH (62-66). In cross-sectional studies, however, the ability to evaluate chronic health effects is limited. In several studies, a positive association between results of one liver function test--the test for y-glutamyltranspeptidase--and PCB blood levels has been found. Kimbrough (67) has summarized earlier studies on the health effects of PCBs observed in workers. In 1930 and 1940. chloracne, a disfiguring skin disease, was reported among workers exposed to PCBs. One of the clinical features of chloracne is the chloracne cyst, which is skin colored and measures from I --10 mm in diameter, wilh a central opening. The other dominant lesion is the comedo. The skin lesions may only involve the face, but many also extend to other parts of the body. Microscopic examination of human skin biopsies from chloracne cases shows markedly dilated hair follicles filled with keratin. The sebaceous glands involute partially or completely. The epithelial cells lining the hair follicles and the adjacent surface epithelium proliferate, and acantho sis is present. In old lesions, the epithelial lining of the greatly dilated hair follicles becomes atrophic. Jones Sc Alden (68) examined 17 of 23 workers engaged in the production of PCBs. The workers had chloracne involving the face, genitalia, trunk, and extremities. Before the outbreak of chloracne in the plant, the electrical properly of the PCBs had fallen below specifications, and the color had deepened. In the report, symptoms of illness were extensively described for the first worker who was diagnosed as having chloracne. This worker com plained of lassitude, loss of appetite, and loss of libido. Over the years, other cases of chloracne following exposure to PCBs have been reported. Most of these involved exposure to vapors that developed when PCBs were healed (69). At times, the skin rashes that developed in workers were accompanied by pruritus. Some workers also complained of burning of the eyes, nose, and throat; dry throat; nausea; and dizziness. Meigs et al (70) reported chloracne in workers who had been exposed to PCB vapors for 5--14 months. The concentration of PCBs in the workers' breathing zone was 0.1 mg/m1. Evidence of slight liver injury was also present. Ouw el al (71) found air levels in a capacitor plant that ranged from 0.32-1.44 mg/m1 Aroclor 1242 (PCB). Here workers complained of burning eyes, face, and skin in general, and persistent body odor. One worker suffered from chloracne, five com plained of eczematous rashes, and a few had abnormal liver function tests. These workers had a mean PCB blood level of about 400 ppb (/xg/kg). In most studies of workers with chloracne, evidence of liver injury was also found; in one study, workers who did not have chloracne were found to have abnormal liver function (69). PCBs also affect the liver by inducing mixed-function oxidases. Alvares et HUMAN HEALTH EFFECTS OF PCBs A PBBs 97 al (72) determined that in five workers occupationally exposed to Aroclor 1016--a PCB mixture primarily composed of dichlnrobiphenyls, trichlorobiphenyls. tetrachlorobiphenyls, and pentachlorobiphcnyls--plasma antipyrine half-life was significantly lower than that in matched controls, suggest ing (he induction of mixed-function oxidases in the liver. These workers had been exposed to Aroclor 1016 for at least two years and had no obvious symptoms of PCB poisoning. Other health effects are eye and upper respiratory irritation. Warshaw et al (63) studied a group of 326 workers in a capacitor plant with a mean employment of more than 15 years and mean employee ages of 41.1 years for males and 47.3 years for females. Work-related eye or upper respiratory irritation was reported by 48% of the workers, and 10% had experienced tightness in the chest. Spiromelric studies were conducted on 309 workers; 66 of them were dropped from the study because (hey had been exposed to talc, textile dust, or asbestos. Thus, 243 men were available for analysis. In males, there were about twice as many smokers and exsmokers as nonsmokers. In females, the proportion of nonsmokers was higher. Thirty-four of the workers (14%) had a reduced vital capacity, and 27 of these demonstrated a restrictive pattern of impairment. Because of additional variables such as smoking and asbestos exposure, these findings arc difficult to interpret. Taylor el al (73), in an attempt to determine whether the fetus would be affected in capacitor workers, examined pregnancy outcome and birth weight, and found (hat the gestation period was reduced by one week. The infants weighed slightly less than the controls; this finding could be explained by the reduced gestation period. Smoking and alcohol consumption were not con trolled, however; furthermore, whether the socioeconomic status of this group of women was similar to that of the control group is not clear. Thus, until other studies confirm these findings, they should be viewed with caution. In several papers Jacobson and his associates reported behavioral changes and a reduced gestation period in association with higher fish intake or higher intake of PCBs (9, 74-76). Furthermore, Jacobson et al (74) reported that intrauterine PCB exposure may have a delayed effect on central nervous system functioning. Since genetic makeup, the mother's lifestyle, and acute illness also affect these parameters, these findings are difficult to interpret. Furthermore, many other chemicals are also excreted in human milk (20). Kogan Sc Gladen (77). for instance, found that mothers with high levels of DDE 11, r-(2,2-dichloroethenylidene)-bis-4-chlorobenzene| in their milk tended to wean (heir infants earlier, as they did not thrive. Apparently, PCB levels in milk were higher in older women, women who drank alcohol regularly, and primiparas (78). Whether high levels of DDE affect lactation is not dear. In animals. DDT tfl: WATER PCB-SD0000012983 98 KIMBROUGH homologs, but not specifically DDE, have been shown to have estrogenic effects (79). Before these findings can be clarified, additional studies must be done. No conclusive evidence thus far reported shows that occupational exposure to PCBs causes an increased incidence of cancer. Bahn el al (80) reponed results of a preliminary study of a group of SI research and development employees and 41 refinery plant employees at a New Jersey petrochemical facility. Between 1949 and 1957 these workers had been exposed to Aroclor 1254. Three melanomas and two carcinomas of the pancreas were found. This incidence was significantly higher than expected. Exposure to other chemicals also occurred, however, and the cohort was small. Brown & Jones (81) conducted a retrospective mortality study of 2.567 workers in two capacitor plants. The relatively few deaths (163) severely limited the statistical power of the study, and the average follow-up was only 15 years, whereas latency periods of 20-30 years are not uncommon for cancer. Over 50% of the sample had exposure to PCBs for two years or less. Deaths from liver cancer, cirrhosis of the liver, and rectal cancer were slightly higher than expected, but not significantly for both sites comhined. The observed increase for cancer of the rectum was statistically significant among females at one of the plants. In a follow up study (82) no additional cancers of the rectum were noted, and the standardized mortality ratio (SMR) dropped from 336 to 211. However, two additional cancers of the liver and biliary tract were observed, bringing the total of these tumors to five as reponed on the death certificates. However, a review of the medical records raises questions about at least one of these tumors. Bertazzi el al (83) reviewed the mortality of 290 males and 1.020 females who had worked for six months or more in capacitor production. Males had a statistically significant increased number of deaths from all neoplasms. When deaths were analyzed by organ system, deaths from neoplasms of the di gestive system, the peritoneum, and the lymphatic and hematopoietic tissues were higher. Among females, all causes of deaths were significantly elevated. The actual numbers in this study, however, were small. Yusho and Yucheng Two outbreaks of poisoning have been reported that followed the ingestion of rice oil contaminated with polychlorinated dibenzofurans, biphenyls, and quaterphenyls (PCQs). The first outbreak occurred in Japan in the summer of 1968 and the second outbreak, in Taiwan in 1979. Ironically, the outbreak in Taiwan repealed what had occurred 10 years earlier in Japan. Many studies of these two outbreaks have been published in Japanese or Chinese. In 1984 some of the information in these reports was published in English in the HUMAN HEALTH EFFECTS OF PCBs & PBBs 99 American Journal of Industrial Medicine 5:1-153; the information is also summarized in volumes 59 and 60 of Environmental Health Perspectives. In Japan and Taiwan the disease was first recognized because chloracne developed in the affected patients (84). In Japan, members of all of the affected households had purchased rice oil from a specific company, and the toxic rice oil produced or shipped on February 5 and 6 of 1968 contained large amounts of Kancchlor 400, a brand of PCB with a chlorine content of 48%. At the time of the outbreak, no analytical methods specific for PCBs were available in Japan; the concentration of Kancchlor 400 in the oil was therefore estimated from the organic chlorine content to be 2,000-3,000 ppm. Kanechlor 400 had been used for heating the rice oil in a metal container at over 200C. at a reduced pressure of 3-44 mm llg, to remove odorous material from it. Kancchlor (K) must have leaked from the heating pipe into the processed oil, but the actual mechanism of the contamination has apparently not been determined. The rcanalysis of the K-rice oil. once the methods were developed, showed that some of the oil samples contained 1.000 ppm (mg/kg) PCB. This concentration was much lower than had originally been estimated. Therefore, other chlorine-containing compounds were assumed to be in the oil. and additional samples were analyzed. The oil was found to contain an average of 5-ppm polychlorinated dibenzofurans (85). According to Buser ct al (86). the Yusho oil contained more than 40 polychlorinated dibenzofuran isomers, including the highly toxic 2,3,7,8-tetrachlorodibenzofuran (TCDF) and 2.3,4.7.8-pentachlorodibenzofuran (PCDF). In addition, the oil con tained PCQs at a concentration of 866 ppm (87, 88). According to estimates made by Kuratsune (89), the total amount of PCB, PCDF. and PCQs consumed by the patients was, on the average. 633 mg of PCB. 3.4 mg of PCDF. and 596 mg of PCQ. This calculates to roughly 157 /ig/kg body weight/d. PCB. 0.9 Mg^g body weight/d. PCDF, and 148 #ig/kg body weight/d. PCQ. At this dose the length of the latent period between exposure and onset of clinical illness was roughly 71 days, with a range from 20 to 190 days. Some of the oil the patients consumed may have contained higher or lower levels because in such situations contamination is usually not uniform. Furthermore, the patients consumed different amounts of con taminated rice oil. The severity of symptoms was positively associated with the amount of contaminated rice oil consumed (24). Early in the outbreak the patients had chloracne. dark-brown pigmentation of the nails, itching, pigmentation of the skin, swelling of the limbs, pig mented mucous membranes, eye discharge, hyperemic conjunclivae, jaun dice. swelling of the upper eyelids, a feeling of weakness, numbness of the limbs, and fever. Over 1.000 people were affected. Thirty-six babies showed fetal PCB syndrome, which consists primarily of a dark-brown pigmentation WATER PCB-SD0000012984 100 KIMBROUGH of (he skin (Cola babies). The cutaneous pigmentation was caused by an increase in melanin pigment in the epidermis (90). The mucous membranes were also pigmented. In all cases, the pigmentation disappeared by the lime the babies were between two and five months old. In affected infants, the face was edematous, and spotty calcifications were noticed in the parietal and occipital areas of the skull. In a few of the infants, the teeth had erupted at birth. Subsequently, the adult patients with clinical disease complained of having to expectorate a great deal and, on auscultation, wheezing was noted; however, on examination, there was no evidence of bronchial asthma or pulmonary emphysema. In many of these patients, the respiratory symptoms have pcrsisled. and the patients have chronically infected airways. In the early 1970s. some changes were noled in the patients' serum immunoglobulin levels, but the levels relumed lo normal. Over lime the severity and the extent of the skin lesions improved considerably in the exposed population. Fifteen yean after the accident, only a very few patients had extensive chloracnc (91). About five yean after the oulbrcak of Yusho, tissue and body fluids of Yusho patients were analyzed for various congenen of PCB and PCDF. At this lime. Ihe PCB levels in adipose tissue were 1.9 1.4 ppm (mg/kg). In Ihe liver Ihey were 0.08 0 06 ppm and in blood, 6.7 5.3 ppb (jxg/kg); thus, they were not very different from levels in the general population in Japan. On the other hand, the isomeric distribution for the PCBs in the Yusho patients varied from that in the control population in the same area (92). About 40 PCDF congeners were identified in Ihe rice oil that the Yusho patients ingested- Only some PCDF congeners were retained in the body for a long lime; they included 2,3,6,8-TCDF, 2.3,7,8-TCDF, 1,2.4,7,8-PCDF, 2.3,4,7,8-PCDF, and 1,2,3,4,7.8-hexachlorinated dibenzofurans. Since these congeners do not have free adjacent carbon atoms, they are not as easily metabolized and excreted. More of the 2,3,4.7.8-PCDF than Ihe other iso mers was retained in ihe patients' tissues. In the five patients studied, the concentration of this isomer ranged from 6.9 ppb (/ig/kg) in a specimen obtained in 1969 to 0.1 ppb (/ig/kg) in a specimen collected in 1977. Measurable concentrations of TCDFs were only detected in Ihe earlier years. Although not the most toxic isomer, Ihe 2.3,4,7,8-PCDF caused mixedfunction oxidase induction at a dose of I /*g/kg in rats, and atrophy of the Ihymus, suggesting toxicity at a very low dosage level. Thus, Ihe clinical o manifestations observed in these patienls were primarily caused hy the PCDFs. specifically by the more toxic isomers. o In the Yucheng episode, il was never determined with certainty how the rice oil was contaminated (93). In 1979 a school for blind persons informed a :) local health bureau in Taichung County lhal a strange disease characterized by 4 an acnelike skin eruption had been occurring frequently among students and v HUMAN HEALTH EFFECTS OF PCBs & PBBs 101 staff since the end of March. At the same lime. 85 of 150 workers in a nearby plastic shoe factory had Ihe same symptoms. Later that year, this outbreak was also reported to a local health bureau. Victims in both outbreaks had consumed the same brand of cooking rice oil. which had been manufactured by the same company and which had been purchased in the same store. For this reason the rice oil was the prime suspect in the oulbrcak. In additional reports of outbreaks in other companies and in the general population, all victims had consumed the same type of C-rice oil. Finally, because Ihe disease resembled the Yusho disease in Japan, samples of C-rice oil and patients' blood were analyzed in Japan and were found lo contain either a Kanechlor-400 or a Kanechlor-500 mixture at concentrations as high as 65 and 108 ppm (mg/kg), respectively. Over 2,000 patients were finally idenlified as having been poisoned by contaminated rice oil. Oil samples collected from other outbreaks contained PCBs at concentrations of 31-300 ppm (mg/kg). Retrospective studies determined lhal ihe period of PCB inlake ranged from 3 lo 9 months. The average total intake for each person varied from 0.77 to 1.8 mg of PCB. Within Ihe first year of the outbreak, the blood levels of PCB in 13 patients ranged from 3 ppb to 1.156 ppb. Most of the patienls had blood levels between 11 and 150 ppb (/ig/kg). The symptoms observed in these patients were quite similar to those already described for the patients in Ihe 1968 Yusho outbreak in Japan. The rice oil was not only contaminated with PCBs but also with PCDFs and polychlorinated quaierphenyls. It contained the same major components of PCDFs observed in the rice oil in Japan--namely. 2,3.4.6.7-PCDF and 2.3.4.7.8- PCDF. Relatively high concentrations of 2.3.4.5.3',4'-hexachlorohiphenyl were found in Ihe blood and adipose tissue of the Yucheng patients. This particular PCB isomer is biologically quite active, and the concentration of 2.3,4,3l4'-pentachlorobiphenyl was also elevated in these patienls. Furthermore, as in the Yusho patients. Ihe concentration of 2.3,7.8TCDF was comparatively low (92). Chen et al (94) analyzed additional samples of ihe oil, blood, and adipose tissue of the Yucheng patients. These investigators identified several TCDF and PCDF isomers. Apparently. 2.3.7.8- TCDF was only a minor component in the oil; the major component was 2.3.4,8-TCDF. One of Ihe major furans in the toxic oil was 2.3.4,7.8PCDF. The concentrations of the PCDFs in different oil samples ranged from 0.21 10 1.68 ppm. Polychlorinated quaierphenyls were present in concentrations ranging from 25 to 53 ppm (mg/kg). Overall, ihe concentrations of PCDFs and PCQs were lower in these oil samples than in the oil samples that had caused Ihe Yusho outbreak. Whether these oil samples were representative is not really known. The 3.4.3'4'-tetrachlorobiphenyl was also identified in the 011 lhal caused Yucheng disease in Taiwan. This isomer is considered to be WATER PCB-SD0000012985 O oo (X) X 102 KIMBROUGH he most loxic PCB isomer present in commercial PCB preparations (95); it was present at a concentration of about 1%. Most other commercial PCB preparations, such as the Aroclors. in the United States have not been shown to contain this particular isomer. In addition to the epidemiological studies, some disease-specific in vestigations were also conducted. The blood pressure of the Yucheng and Yusho patients was not affected (60). Although some of the patients in the Yusho cohort have died of cancer (90). the number has been small; because the latency period may be long, the population should be followed for a longer period to determine whether Ihe cancer incidence will increase. Although PCBs and related compounds are known to affect reproduction in animals, and although they affected some fetuses and neonates in the Yusho and Yucheng episodes, the information on reproduction and fetal toxicity in general is very limited. In one such study, Hara (96) examined women working in a capacitor plant who also nursed their infants and who themselves had mild chloracne and erythema of the skin. The human milk of some of these women contained, on a whole milk basis. PCB levels that ranged from below 50 ppb (/xg'kg) to about 400 ppb (/xg/kg). Forty children of these mothers were followed for a five-year period. Some children were found to have "decayed" nails, gingival pigmentation, mottled enamel, and denial caries. No relationships between these changes or symptoms to PCB blood levels, however, were observed. The general population in the United States and other countries also has body burdens of PCBs, PCDFs. and polychlorin ated dibenzodioxins (97, 98). However, these background concentrations-- particularly for the biologically active isomers--are far lower than they were in the Yusho and Yucheng patients, even several years after exposure. Chang el al (99) examined the delayed immune response in 30 Yucheng patients and compared their responses with those of 50 controls. The mean age of patients in both groups was about 14 years. The authors injected a solution of streptokinase and streplodomasc subcutaneously into the flexor side of the forearm. The response was read al 24 hours (hr) and again at 48 hr after injection. Eighty percent of Ihe controls had an induration of 5 mm or more in diameter 24 or 48 hr after they were injected; only 43% of the exposed group responded similarly. All of the poisoned patients had dermal lesions, and Ihe percentage of patients with a positive response decreased with increasing severity of the skin lesions (chloracne). Furthermore, the degree of the dermal lesions appeared to be associated with the whole blood PCB concentrations. Patients with minor skin lesions that were classified as grade I appeared to have a normal skin response. The same authors found that PCBs caused a decreased concentration of IgA and IgM. but not of IgG, in serum. Furthermore, the percentages of total T cells, active T cells, and T mu cells decreased, whereas Ihe percentage of B cells and T gamma cells were not HUMAN HEALTH EFFECTS OF PCBs & PBBs 103 affected (100). These two reports are the first in which the effect on the immune response was actually correlated with body burdens of PCBs and in which only severely poisoned patients showed this effect. This finding is consistent with the findings from animal studies in which relatively high doses of PCBs affected the immune response and also caused some other adverse effects. In the Japanese and Ihe Taiwanese Yusho and Yucheng poisoning out breaks, sensory neuropathy was reported in a number of patients for whom nerve conduction velocities were measured (101, 102). The blood levels of Ihe various chemicals (PCBs. PCDFs, PCQs) were negatively correlated with the lowered nerve conduction velocity, suggesting that these types of chemi cals affect nerve conduction velocity. (If is not quite clear why most in vestigators measure nerve conduction velocity to detect sensory neuropathy. Other tests that would measure the detection of vibration, touch, and tempera ture would be more useful from a clinical perspective.) Seppalainen el al (103) examined 16 men working in a cardboard plant who were exposed to fumes that resulted from the explosion of 15 capacitors containing Clophen A-30. The first PCB air concentrations, measured 5.5 hr after the explosion, were 8,000 to 16,000 /xg/m3 air. PCDFs were also formed. The soot samples contained tetrachlorodibenzofuran up to 90 /xg/g. of which 6.5 /xg/g was 2,3.7.8-tetrachlorodibenzofuran. In addition, monochloropyrenes and dichloropyrenes were found. Most of the men had a transient sensory neuropathy in their lower extremities. Chang et al (104) reported increased urinary 5-aminolevulinic acid uropor phyrin excretion in 69 Yucheng patients over that of 20 controls. No informa tion on the patients' clinical conditions or on how these findings related to degree of exposure was given. No such observations have been reported from Japan. POLYBROMINATED BIPHENYLS Since the toxicity of PBBs both in laboratory animals and livestock was recently reviewed (105), we do not review it here in detail. In laboratory animals, PBBs generally cause effects similar to those that the PCBs cause. They produce morphological changes in the liver, affect reproduction, and promote biochemical changes, such as hepatic porphyria and induction of mixed-function oxidases. Teratogenic effects have also been noted. In addi tion, atrophy of the thymus has been reported, and hepatocellular carcinomas have been produced in both rats and mice. The overall findings reported in animal studies are similar to those that have been reported for PCBs. Although the PCB contamination of the environment is a more general problem, Ihe PBB contamination primarily affects certain areas within the state of Michigan. Most persons living within the lower peninsula of Michi- WATER PCB-SD0000012986 104 KIMBROUGH gan have had slight exposure, since the contamination resulted from dairy products and since normal marketing channels for these products involved the mixing of milk from many producers in relatively few processing facilities. In addition, most cull dairy cattle are used for hamburgers and processed meat products that would also receive wide distribution. Thus, the marketing system diluted the degree of exposure for the individual; however, it increased the number of those exposed. In 1978, the distribution of PBBs was com prehensively studied in a probability sample of 1.738 persons. PBB levels in scrum were determined, and 844 adipose tissue samples were also analyzed for PBBs. PBBs were detected in 97.3% of the adipose (issue samples, in 68% of the adult serum samples, and in 72.7% of the serum samples from children. The mean PBB concentration in adipose tissue was 400 pph (/ig/kg); in scrum it was 1.3 ppb (/ig/kg) for adults and 1.8 ppb (ju/gkg) for children. The highest adipose tissue concentration was 37 ppm (mg/kg) (18). In addi tional studies, when cohorts of PBB-exposed residents of Michigan were compared with residents of the stale of Wisconsin, a higher prevalence of a variety of symptoms and complaints was noted in the Michigan residents (106). Similarly, in comparative neurobehavioral studies, the Michigan pop ulation was found to be affected more than that in Wisconsin (107). Since the findings were not correlated with body burdens of PBB in any of these studies, determining whether other factors may be responsible for these differences is difficult. In 1976, the Michigan Department of Public Health established a cohort of farmers who had been exposed to varying con centrations of PBBs in their products and their environment. A total of 3.877 persons were enrolled. They included farm residents, direct recipients of farm products, chemical workers and their families, and a few persons who had been originally studied in a smaller previous study. The serum PBB levels in this entire group ranged from no detectable levels to 1,900 ppb (pg/l). with a mean of 21.2 ppb (pg/l) and a median of 3 ppb (jsg/1). Because of (he wide range of exposure and because results could be analyzed by regression analyses with exposure as a variable, a comparison group for acute health effects was not included. This cohort was found to have various symptoms and conditions; however, these symptoms did not correlate with PBB body burdens. Symptom prevalence rates were slightly higher in persons with no detectable PBBs in serum than in those with measurable quantities. In all groups, including chemical workers and quarantined farm residents, the highest prevalence rates were in persons with the lowest serum PBB levels (22). Similarly, in this study and in a previous immunologic study (108) no dosc-relaled depression of lymphocyte function in persons exposed to PBBs could be demonstrated. All these findings suggest that there may be no causal HUMAN HEALTH EFFECTS OF PCBs & PBBs 105 relationship between the abnormal lymphocyte functions observed in some persons or the prevalence of other symptoms and exposure to PBBs. This cohort of Michigan residents is still being followed by the Michigan Depart ment of Health in collaboration with the Centers for Disease Control. Several studies of subgroups of this population pnd surveys for chronic health effects have been conducted since the cohort was First assembled (109, 19). When serum and adipose tissue concentrations were compared, a significant correla tion was found. The serum: adipose tissue concentration ratios ranged from I to 140 to I to 260 for pregnant women and male chemical workers, respec tively. Males from farms had a significantly different ratio of I to 325 (o 329. Potential transplacental passage of PBBs was demonstrated, since they could also be found in the fetus and newborn. Cord blood contained one-tenth of the concentration found in the maternal serum, which indicated partial placental passage. Human milk contained PBBs at 107-119 times the quantity found in maternal serum. PBBs were also delected in bile and feces, which indicates that these materials can be transferred into the intestinal tract. All of these concentrations were measured long after the population bad first been exposed to PBBs (19). Concentrations of PBBs observed in bile and feces were about one half to seven-tenths of the serum levels and are probably about 0.5% of (he adiposejissue levels. These findings indicate lhai PBBs are very slowly excreted, which is consistent with the Findings of Tuey &. Matthews in rats (110). The estimated half-life for PBB is 6.5 years. More recently, two groups of Michigan residents--those with high PBB serum levels and those with PBB serum levels around I ppb--were matched for age. sex, and smoking. For both groups, various clinical laboratory tests were conducted, blood pressure was measured, and height and weight were determined. In this study. 83 participants had PBB serum levels of 50 ppb (/x.g/1) or more. In the middle group, 83 had PBB levels of 5-49 ppb and 96 had PBB levels of 0-44 ppb (/x.g/1) in serum. Urinary porphyrins were also measured in all of the participants. Thus far. the final results of this study have not been reported. For most of the parameters studied--which included serum glucose, triglycerides, high-density lipoproteins, various liver func tions, creatinine, uric acid, thyroid function, proteins, calcium and phospho rus in serum, and also measurement of various porphyrins--no differences of clinical significance were found among different groups (M. Barone. personal communication). (Note: Even though significantly more women in the high PBB group used birth-control pills than women in (he low PBB group, too few were using them to affect urine porphyrin levels.) In none of a variety of other studies conducted on (his population as well as other groups in Michigan did any Findings indicate that exposure to PBB had impaired the health of the exposed group. All of these studies have been reviewed by Fries (105). WATER PCB-SD0000012987 106 KIMBROUGH Allhough this population was exposed during a 9-monlh period in 1973 and 1974. whether ii will have chronic health effects is unknown. This particular cohort needs lo be followed for 30 lo 40 years before the question of chronic health effects can be intelligently addressed. Two problems with assessing chronic healih effects are that the cohort, in spite of iis size, is slid relatively small and lhat ihe amount of exposure it has received varied widely. Although some members of Ihe group exposed lo PBBs have relatively high body burdens, these burdens are still appreciably lower than those of rats in which liver cancer developed. In the study by Kimbrough el al (111), liver cancer developed in the rats lhal received a dose of 1,000 mg/kg body weight. This dose for humans would roughly translate into a dose of 70 grams per person. These amounts are much greater than the estimated mean total exposure per person. The highest exposure was about 11.7 grams, and Ihe mean was 170 mg per person. In rats given 200 mg/kg. a dose lhal for humans would be between 12 and 14 grams, only neoplastic nodules developed in their livers: there was no evidence of hepatocellular carcinomas. Of course, whether humans would be more or less susceptible lo the toxic effects of PBBs and whether their response would be similar to that of rats is not known. In conclusion, various toxic effects of PBBs and PCBs have been described in laboratory animals. In humans, acuie poisoning outbreaks have only occurred following exposure lo a combination of PCBs and PCDFs. When humans were exposed only lo PCBs or PBBs. Ihe only observed acute effects have generally been minor. So far. no significant chronic healih effects have been causally associated with exposure lo PCBs or PBBs. Use of trade names is for identification only and does not constitute endorsement by Ihe Public Healih Service or Ihe US Department of Health and Human Services. Literature Cited 1. Jensen. S. 1966 Report of a new chernicl hazard. New S* i. 32:612 2. 19X2. Polychlorinated biphenyls (PCBs): manufacturing, processing, distribution in commerce, and use prohibitions: use in electrical equipment. Fed, Mrgin. 47:37342-6(1 3. |982. Polychlorinated biphenyls (PCBs): manufacturing, processing, distribution in commerce, and use prohibitions: use in dosed and controlled waste man* ufacturing processes. Fed. Regal, 47:469X0-96 4 McLeod. K E 1981 Polychlorinated biphenyls in indoor air. Environ. Sd. Tethnal. 15.926-28 5. Kuiz, F. W.. Vang. H. S C 1976 A note on polychlorinated biphenyls in air. In Pros'. Natl. Can/. Paivchlorinnted Binhenvh. Chicago. 1975. |:PA 360/6 75-004. p. 182. Washington. DC: En viron. Prof. Agency 6. Cordle, F., Locke. R.. Springer. J. 19X2. Determination of human risk in regulating polychlorinated biphenyls (PCIIst--a case study. CRC toiiiol. Rttk Attest. 2 211 25 7. Curdle. F.. Locke. R.. Springer. J. 1982. Risk assessment in a federal reg ulatory acency: an assessment of risk associated with the human consumption of some species of fish contaminated with polychlorinated hiphcnyls (POBsl. Lm'iro/i Henilh Persped. 45.171-82 I t i I HUMAN HEALTH EFFECTS OF PCBs A PBBs 107 8. Sullivan. J. R.. Delftno. J. J., Buelow, C. R.. Sbcffy. T. B. 1983. Polychlori nated biphenyls in the fish and sediment of Ihe Lower Fox River. Wisconsin. Bull. Environ. Contam. Toxicol. 30:58 64 9. Schwartz. P. M.. Jacobson. S. W.. Fein. G. G.. Jacobson. J. L.. Price. H. A. 1983. Lake Michigan fish consump tion as a source of polychlorinated biphenyls in human cord serum, mater nal serum, and milk. Am. J. Public Health 73:293-96 10. Tatsukawa. R. 1976. PCB pollution of Ihe Japanese environment. In PCB Poisoning and Pollution, cd. K. Higuchi. pp. 147-79. Tokyo: Kodansha 11. Morita. M.. Nakagawa. J.. Rappe. C. I97H. Polychlorinated dibenzofuran (PCDFl formation from PCB mixture by heat and oxygen. Bull. Environ. Contam. Tosicol. 19:665-70 12. OKeefe. P. W. Silkworth. J. B.. Gierthy. J. F.. Smith. R. M.. DeCaprio. A. IV. et al. 1985. Chemical and biological investigations of a transformer accident at Binghamton. New York. Environ. Health Perspect. 60:201-9 13. Hutzinger. O., Ghuiam, G. C., Brock. Ci. C.. Johnston. I.. E 1983. Formation rtf polychlorinated dibenzofuran* and dioains during combustion, electrical equipment fires and PCB incineration. Environ. Health Perspect. 60:3-9 14. Carter. 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