Document 2R8YjM6QBjm7dr9rr2gXOLjg5

- P90>J(T no: ' RtfOR, FILE ( 8 SfT$u&t/ufiJ BI O - T E S T 1810 FRONTAGE ROAD NORTHBROOK, ILLINOIS 60062 3no. vy REPORT TO MONSANTO COMPANY THREE-GENERATION REPRODUCTION STUDY WITH AROCLOR 1242 IN ALBINO RATS NOVEMBER 1, 1971 IBT NO. B7297 ( ( 0365389 TOXSTUDIES1147 9ru!uit/u<il B i o - t e s T XdxmkyiM'. 9nc. 1810 FRONTAGE ROAD NORTHBROOK, ILLINOIS 60062 November 1, 1971 />// ;v-<. Mr. Elmer Wheeler Monsanto Company 800 N. Lindbergh Boulevard St. Louis, Missouri 63166 Dear Mr. Wheeler: Re: 1BT No. B7297 - Three-Generation ReproductionStudy With Aroclor 1242 In Albino Rats We are submitting herewith our laboratory report dated November 1, 1971, prepared in connection with the above study. Very truly yours, JCC:sjn J. C. Calandra President 0365390 TOXSTUDIES1148 'tindathtel B I O - T E S T latKyiabyu&i, 9hc. ( i REPORT TO MONSANTO COMPANY THREE-GENERATION REPRODUCTION STUDY WITH AROCLOR 1242 IN ALBINO RATS NOVEMBER 1, 1971 IBT NO. B7297 I. Introduction At the request of Monsanto Company, a reproduction study was \. ( initiated in order to ascertain potential toxicological effects of subacute oral administration of Aroclor 1242, Lot No, AK-255, upon the fertility and offspring of three consecutive generations of albino rats. This report presents the data collected from the initiation of the investigation to the weaning of the second (F3b) litters obtained from the third (F2) parental generation. C 0365391 TOXSTUDIES1149 BIO-TEST ab>'iaifr\iei-l 9nc. Z II. Summary A three-generation reproduction study employing albino rats fed diets containing 1, 10 or 1 00 ppm Aroclor 1242 was conducted. The fol lowing results were obtained during the investigation. A. Parental Animals 1. Body Weights Body weights and body weight gains revealed no differences between test and control animals which could be attributed to the ingestion of Aroclor 1242. 2. Mortality and Reactions Several animals succumbed during each of the three generations. None of these deaths could be attributed to the ingestion of Aroclor 1 242. 3. Pathologic Studies Gross pathologic examination revealed no differences between test and control rats. The absolute liver weights and liver to body weight or brain weight ratios were significantly elevated in both FI males and females fed 100 ppm Aroclor 1242. Statistical evaluations of other organ weights, organ to body weight and organ to brain weight ratios disclosed several randomly occurring intergroup differences. The lack of any consistent dose or generation related response and the absence of any deleterious histopathologic change confirm that these differences were not related to the ingestion of Aroclor 1242. TOXSTUDIES1150 9ndMsUal B I O - T E $ T latMaivUet, 9*tc. 3 All lesions noted during histopathologic studies were those of spontaneous disease. 4. Reproductive Performance Reproductive indices of rats fed 1 or 10 ppm did not differ from those of the controls during any of the three generations. During the second generation (F2a and F2b litters), rats fed 100 ppm had lowered mating indices and a reduced incidence of pregnancy. Because only two T-III females produced F2b litters, it was not possible to continue this level through the third generation. B. Progeny 1. Population Data The number of live pups born per litter and the avejage number of pups weaned were not affected at any level of Aroclor 1242 tested. No external abnormalities were present that could be attributed to treatment. No untoward behavioral reactions were observed among test or control progeny. 2. Survival Twenty-four hour and five day survival indices and the lactation index were normal for all groups. 3. Body Weights Body weights for all progeny were normal at weaning. 0365393 TOXSTUDIES1151 t . 9nduiUial B I O - T E $ T 2ahviaiiyu&i, 9nc. 4 ( 4. Pathologic Studies At autopsy of the F3b weanlings no gross or microscopic tissue changes were present which could be associated with the ingestion of Aroclor 1242. Respectfully submitted, INDUSTRIAL BIO-TEST LABORATORIES, INC. Report prepared by: Sandra Haley Assistant Toxicologist Rat Toxicity / Report approved by: Jhjfnes B. Plank Senior Group Leader Rat Toxicity /^/^^_ Paul L. Wright, *Pn. D. Section Head, Toxicology November 1, 1971 Jg:6jn;psh M. L, Kepling^r, Ph.^T Manager, Toxicology ( 036539* TOXSTUDIES1152 k ftufo&kiol BI O T E S T 2olMafo>uM., 9nc. 5i HI. Procedure A. Experimental Animals The animals employed were weanling Charles River albino rats*. A total of 96 weanling rats (32 males and 64 females) was selected to form three test groups and a control group of Fo generation animals. Parental animals were allowed to reach maturity, mate and produce two litters. Eight males and 16 females from the second litters of each dietary group (Fib and F2b) were selected at weaning as parental animals for the succeeding generations. The study terminated following the weaning of the F3b litters. B. Organization of Groups A structural outline of the experiment is given in Table I. A flow chart depicting the three generations is shown in Figure 1. * Charles River Breeding Laboratories, Inc., North Wilmington, Mass, 0365395 TOXSTUDIES1153 'SnduiUial BIO-TEST J-ctltfr'ialasti&i, 9nc. 6 ( TABLE I TEST MATERIAL: Aroclor MM Three-Generation Reproduction Study - Albino Rats Outline of Experiment G r oup Dietary Level (ppm) Number of Animals Male F emale Control None 8 16 T-I 1 8 16 T -II 10 8 16 T -III 100 8 16 ( C T0XSTUDIES1154 ,, ` # StidnsUial BIO-TEST dalonatMieZ, 9nc. 1\ 7 FIGURE 1 TEST MATERIAL: Aroclor 1242 Design of Experiment Fo Parental Animals G e n e ra tio n G e n e ra tio n G e n e ra tio n F irs t Fla Progeny Wean Sacrifice Fib Progeny Wean Select the FI Parental Animals Second F2a Progeny Wean Sacrifice F2b Progeny Wean Select the F2 Parental Animals ( F3a Progeny F3b Progeny Wean W ean Sacrifice Histopathology T h ird Gross and histopathologic examinations were conducted upon parental animals of all three generations following the weaning of the second (b) litters. C T0XSTUDIES1155 , S(<n4u4^uai B I O - T E S T JaiotefoueA, 9nc. 8 / C. Diets and Feeding Diets were prepared in the central diet room of this laboratory. The diet for any given test group was prepared by incorporating a calculated weight of Aroclor 1242 into a standard pulverized stock ration*, so that the final concentrations of test material in the diets were 1, 10 and 100 parts per million respectively. The animals in the control group were fed the basic ration without test material. Animals in all groups were maintained on their respective diets without interruption until their sacrifice, which followed the weaning of the second litters. Feeding was conducted on an ad libitum basis with fresh, diets prepared weekly, D. Body Weights and Weight Gains ^ Initially, the body weight of each rat was determined and recorded at 21 days of age. All animals were then weighed weekly until mating trials commenced. No further body weight data were obtained until the sacrifice of each parental generation, at which time final body weights were recorded. E. Mortality, Reactions and General Observations Observations among animals of each generation for mortality and abnormal behavioral reactions were made daily. In addition, these animals were observed for fertility, length of gestation and lactation performance, * Special Mix Mouse Chow, Ralston Purina Company, St. Louis, Missouri, (' ' 0365398 TOXSTUDIES1156 . fautuifaial B IO ** T E S T $nc. 9 ^ All pups were examined for physical abnormalities at birth and the numbers of viable and stillborn members of each litter were recorded. Records of survival at designated intervals during the lactation period were maintained and a final examination for physical abnormalities was made at the weaning of each litter. Litters of more than ten pups were reduced to that number by random sacrifice on the fifth day of the lactation period. F. Mating Procedure Mating trials were initiated when the parental animals were 100 days old. Females were caged in pairs and mated with a male from within the same treatment group. Daily examinations were conducted ^ to determine if copulation had occurred. Males were rotated within their dietary group at ten day intervals until conception was confirmed or until each female had been paired with a maximum of three males. The first litters (Fla, F2a and F3a) obtained were weaned at 21 days post-partum. The parental females were then given a ten-day rest period and again mated, the above procedure being repeated in order to obtain second litters (Fib, F2b and F3b). Females failing to become pregnant during the first mating trials were not rebred. Eight males and 16 females from the second litters of each group were selected at weaning on the basis of health and body weight as parental animals for each succeeding generation. C 0365399 TOXSTUDIES1157 'fyidui&iial B 1 O - TEST Ja&otaUiict. $nc. 10 C, Gross and Microscopic Pathologic Studies Whenever a parental animal succumbed during the test period, a gross autopsy was performed. In those instances when postmortem changes were not advanced, representative tissues were taken for microscopic study. After the second litters were weaned, all surviving males and eight feinales from each group were sacrificed and subjected to gross pathologic examination. The weights of the liver, kidneys, spleen, gonads, heart and brain were recorded, along with the final body weight of each rat. Statistical analyses were conducted upon the absolute organ weights of the sacrificed animals and upon the corresponding organ to body weight and organ to brain weight ratios. An Analysis of Variance was conducted first and significant effects disclosed by that treatment were further studied by 'V-tests. Microscopic pathologic studies wore conducted upon five males and five females (Fo and FI) from the control and T-III groups; the T-II and control groups were examined following sacrifice of the F2 generation. The following tissues and organs were examined: 0365*00 TOXSTUDIES1158 faufultual B I O - T E S T Ja&MafWte, 9nc. ( Tissues and Organs ' Examined Grossly Skin Heart Trachea Lungs Liver Pancreas Esophagus Stomach Intestinal Tract Spleen Lymph Nodes Kidneys Urinary Bladder Testes Ovaries Prostate Seminal Vesicles Uterus Vagina , ( Pituitary Gland Adrenal Glands Salivary Glands Thyroid Gland Skeletal Muscle Bone Peripheral Nerve Eyes Brain Thymus Optic Nerve Aorta 11 Tissues and Organs Examined Microscopically Heart (right and left ventricles) Trachea Lung Liver Pancreas . Stomach (cardiac, fundic and pyloric regions) Small Intestine (duodenum, jejunum and ileum) Caecum Colon Spleen Lymph Node (cervical and mesenteric) Kidney Urinary Bladder Testis Ovary Prostate Uterus Pituitary Gland - Adrenal Gland Salivary Gland (submaxillary) Thyroid Gland Parathyroid Gland Skeletal Muscle Bone Marrow (sternum and femur) Peripheral Nerve (sciatic) Brain (cerebrum, cerebellum and pons) Seminal Vesicles Esophagus Spinal Cord t 0365401 TOXSTUDIES1159 UndutUial BIO - T E S T alxyiaio>iiel. 9nc. *^ iv. Results A. Parental Animals 1. Body Weights Body weight data are presented in Tables II and III. Included in the data on these tables are total weight gains over the first 77 days of the feeding period. There were no differences between test and control animals which could be related to the ingestion of Aroclor 1242. j i ( C 3654o^ TOXSTUDIES1160 B IO - T E S T lalt&xd&u&i-, oup Generation 0 Fo FI 47 F2 49 I ppm) ' Fo F1 F2 48 58 -II Fo 0 ppm) FI 50 F2 52 -HI 00 ppm) Fo FI 45 TABLE II TEST MATERIAL: Aroclor 1242 Three-Generation Reproduction Study - Albino Rats Parental Male Rats Mean Body Weight (grams) Day Number: 7 14 21 2S 35 42 49 56 63 70 77 95 142 202 253 309 354 393 47.6 456 480 492 77 112 152 192 234 271 316* 349 382 416 438 77 108 140 164 200 234 265 295 319 345 354 95 137 192 242 292 336 374 409 435 458 475 76 115 164 212 259 297 335 371 393 421 444 98 149 195 234 280 327 357 390 408 436 455 95 141 200 249 297 341 366 394 422 447 466 85 123 169 221 271 306 346 394 422 438 455 92 136 176 221 262 285 313 341 362 380 394 95 139 196 224 292 340 375 400 433 461 474 73 112 153 194 245 291 339 366 409 432 450 Total Weight Gain (grams) Final Body Weight at Sacrifice 397 391 305 709 636 592 380 396 397 707 639 680 371 405 342 661 636 574 379 405 720 652 ote: All animals were 28 days old at day number seven. T0XSTUDIES1161 M u i h l a l B I 0 - T E ST Is th v d w ie i, }nc. TABLE III TEST MATERIAL: Aroclor 1242 Three-Generation Reproduction Study - Albino Rats Parental Female Rats Mean Body Weight (grams) Day Number: rup Generation 0 7 14 21 28 35 42 49 56 63 70 77 jpm) I ppm) in 0 ppm) Fo FI F2 Fo FI F2 Fo FI F2 Fo FI 93 127 158 182 203 220 233 240 250 257 266 45 73 104 134 156 180 197 220 229 237 246 248 44 66 89 113 134 152 163 178 183 193 199 212 . 93 126 155 179 203 221 232 243 252 262 270 4b 75 107 140 156 176 196 216 225 229 248 252 48 76 104 134 144 169 177 186 191 205 213 214 . 93 127 158 181 200 219 230 241 248 256 263 49 78 107 138 167 176 204 222 237 249 256 263 49 79 116 137 160 177 189 200 212 223 231 237 . 93 127 160 184 207 224 236 247 254 264 272 42 69 103 136 160 182 206 223 237 252 261 270 iet All animals were 28 days old at day number seven. Total Weight Gain (grams) Final Body Weight at Sacrifice 173 203 168 388 349 316 177 206 166 388 304 318 170 214 188 387 338 360 179 228 372 392 w O\F' TOXSTUDIES1162 jhuL&t'ual B I O - T E S T Jtabonaivu&i, $*ic. 15 2. Mortality ' Mortality data are presented in Tables IV and V. Gross autopsy performed on animals which succumbed during the test period failed to reveal any relationship between the ingestion of Aroclor 1242 and the death of the animal. 0365405 TOXSTUDIES1163 1 S 3 - O I 3i w$ TABLE IV TEST MATERIAL: Aroclor 1242 Three-Generation Reproduction Study - Albino Rats Mortality Data Parental Generation Males Group Generation C Fo FI F2 T-I (1 ppm) Fo FI F2 T -II (10 ppm) Fo FI F2 T -III Fo (100 ppm) FI r'irst Mating Pre-M M Post-M 000 000 000 000 0 0 0 000 000 000 000 1 00 000 Second Mating M Post-M Total 000 000 01 1 000 01 1 000 000 0 0 -o 01 1 00 1 0 1 .1 Pre-M = Pre-Mating Period M = Mating Period Post-M = Post-Mating Period TOXSTUDIES1164 lutbUol BI 0 - T E S T JtaUvJaMol, 3m. TABLE V TEST MATERIAL: Aroclor 1242 Three-Generation Reproduction Study - Albino Rats Mortality Data Parental Generation Females Group Generation C Fo FI F2 T-I (1 ppm) Fo FI F2 T-II (10 ppm) Fo FI F2 T -III Fo (100 ppm) FI Pre-M 0 0 0 0 0 0 0 0 0 0 0 First Mating MG P 001 000 000 000 000 000 000 000 000 000 000 Pre-M = Pre-Mating Period M = Mating Period G = Gestation Period P = Parturition L = Lactation Period R = Resting Period Post-W = Post-Weaning Period LR 00 00 01 0 .0' 00 00 00 00 00 00 00 Second Mating M G 'P L Post-W T otal 00000 1 000000 000001 0001 01 00001 1 00 00 00 000000 00 0 000 000000 000000 000000 40VS9C0 TOXSTUDIES1165 B I O - T E S T Jialvu$no. 18 3. Reactions No untoward reactions were noted among the parental ani mals during the investigation. 4. Pathologic Findings a. Gross Pathologic Findings There were no gross pathologic findings which could be correlated with ingestion of the test material. Findings among test animals were comparable to those noted among control animals. b Organ 'Weights and Ratios Mean organ weights, organ to body weight ratios and organ to brain weight ratios are assembled in Tables VI through XIII, The absolute liver weights and liver to body weight or brain weight ratios were significantly elevated in both FI males and females fed 100 ppm Aroclor 1242. Statistical evaluations of the other organ weights and ratios revealed several intergroup differences. There was no consistent dose or generation related response and these differences were not attributed to the ingestion of Aroclor 1242. - TOXSTUDIES1166 B 1O - T E S T Ja&oAaioAiet., 9*- CO CO Group Generation C. To FI F2 T-I (1 ppm) Fo FI F2 T-II (10 ppm) o o- T-III (100 ppm] c Fo FI F2 Fo FI TABLE VI TEST MATERIAL: Aroclor 1242 Three-Generation Reproduction Study - Albino Rats Mean Organ Weights and Ratios Parental Animals Organ: Liver Organ Weight (g) Males F emales 28. 0 24.7 23. 6 16.3 15. 2 12.2 Organ/Body Weight Ratio (g/100 g) Males Females 3. 96 3. 90 3.97 4.26 4.41 3.87 Organ/Brain Weight Ratio (g/g) Males Females 12. 9 11.9 11.6 8. 68 7. 92 6. 97 00 26.8 24.4 22.7 26.5 26. 7 22. 5 16.8 13.4** 12.0 16. 0 16. 1 15. r 3. 80 3. 87 3.38** 4. 02 4. 20* 3. 94 4.33 4.45 . 4. 18 4. 78 4.25 . 12. 6 11. 8 10. 5 12. 0 -12.4 10. 6 7. 06* 6. 02 7. 77 8. 36 7. 80 29.9 30. 8* 17. 6 18.3** 4. 18 4.75* ` 4.75 4.73 14.4* 15.8** 7. 90 10.30** * Statistically significant difference at the 95 percent confidence level. ** Statistically significant difference at the 99 percent confidence level. TOXSTUDIES1167 `iitwfDWfor l s 3 i o 13 Group Generation C Fo FI F2 T-I (1 ppm) Fo FI F2 T -II (10 ppm) T-m (100 ppm) Fo FI F2 Fo FI TEST MATERIAL: Aroclor 1242 Three-Generation Reproduction Study - Albino Rats Mean Organ Weights and Ratios Parental Animals Organ: Kidneys Organ Weight (g) Males Females 4. 82 4.44 4. 01 2. 77 2. 68 2. 43 4. 30 4.47 4. 03 2. 72 2. 65 2. 37 4. 59 4. 56 3. 71 2. 95 2. 74 2. 68 4. 26 4. 09 3. 02 2. 74 Organ/body Weight Ratio (g/100 g) Males ' Females Organ/Brain Weight Ratio (g/g) Males Females - 0. 680 0. 709 0.681 0. 724 0. 771 0. 774 2. 22 2. 14 1.99 1.48 1.40 1. 39 0.610* 0. 709 0.601** 0.704 0.882** 0.755 . 2. 02 2. 17 1.87 1.39 1. 39 1. 19 0. 693 0.723* 0. 647 0. 771 0. 813 0. 755 2. 08 2. 13 1.74 1.44 1.42 1. 38 0.593** 0. 6?6* 0.816** 0.711 2. 05 2. 09 1.33 1. 55 * Statistically significant difference at the 95 percent confidence level, ** Statistically significant difference at the 99 percent confidence level. TOXSTUDIES1168 Itia i B I O - T E S T Jobvuifoii& l, $nc. Group Generation C Fo FI F2 T-I (1 ppm) Fo FI F2 T-H Fo (10 ppm) o FI Cf F2 t-iii (100 ppm) Fo FI TABLE VIII TEST MATERIAL: Aroclor 1242 Three-Generation Reproduction Study - Albino Rats Mean Organ "Weights and Ratios Parental Animals Organ: Spleen Organ Weight (g) Males Females 0. 83 0. 74 0.96 0. 63 0. 61 0. 63 0. 85 0.73 0.83* 0. 89 0. 78 0.76* 0. 59 0.53 0. 57 0. 59 0. 64 0. 62 . 0. 82 0.82 0.58 0. 60 Organ/Body Weight Ratio (g/100g) Males ' Females 0. 12 0. 12 0. 16 0. 16 0. 18 0. 20 0. 12 0. 12 0.12** 0. 14 0. 12 0. 13* 0. 15 0. 18 0. 18 0. 15 0. 19 0. 18 0. 11 0. 13 ' 0.16 0.15 Organ/Brain Weight Ratio (g/g) Males F emales 0. 38 0. 36 0.48 0. 33 0. 32 0. 36 0.40 0. 35 0.38** 0.41 0. 36 0.35** 0. 30 0. 28 0. 28 0. 29 0. 33 0. 32 0.40 0.42 0. 25 0. 34 * Statistically significant difference at the 95 percent confidence level. ** Statistically significant difference at the 99 percent confidence level. TOXSTUDIES1169 BIO -TEST j* Group Generation C- Fo FI F2 T-I (1 ppm) Fo FI F2 T -II (10 ppm) Fo , c FI O vn F2 T-IH (100 ppm) Fo FI TEST MATERIAL,: Aroclor 1242 Three-Generation Reproduction Study - Albino Rats Mean Organ Weights and Ratios Parental Animals Organ: Gonads Organ Weight (g) Males F emales 3. 62 3. 57 3. 18 0. 117 0. 099 0. 132 Organ/Body W eight Ratio (g/100 g) Males ' Females 0. 510 0. 576 0.530 0.0302 0.0283 0. 0424 Organ/Brain Weight Ratio <g/g) Males Females 1. 67 1.72 1.57 0.0627 0.0524 0.0757 3. 59 3. 66 3. 56 0. 116 0. 080 0. Ill 0. 512 0. 581 0. 536 0. 0303 0.0263 0.0353 1.69 1.77 1.65 0.0590 0.0415 0.0561 . 3. 61 3. 78 3. 74 3. 55 3. 69 0. 131 0. 119 0. 130 0. 114 0. 102 0. 549 0. 630 0.652* 0.0338 0.0361 . 0.0375 0. 501 0. 571 ' 0.0310 0.0274 1.64 1.76 1.75 1. 70 1.89 0.0637 0.0620 0.0668 0.0505 0.0579 * Statistically significant difference at the 95 percent confidence leve! tv tv TOXSTUDIES1170 B I O " T E S T XatuyiakviieA, ?*ic. Group Generation C To FI F2` T-I (1 ppm) Fo FI F2 t-ii (10 ppm) Fo FI O' F2 T-m (100 ppm) Fo FI TABLE X TEST MATERIAL: Aroclor 1242 Three-Generation Reproduction Study - Albino Rats Mean Organ Weights and Ratios Parental Animals Organ: Heart Organ Weight (g) Males Females I. 36 1.74 1.34 1.28 1.24 1. 19 1.79 1.87 1.93 1.28 1.32 1. 12 1. 71 1. 82 1. 57 1.92 1. 83 1.25 1.10** 1. 18 1.25 1. 28 Organ/Body Weight Ratio (E/100 g) Males ' Females Organ/Brain Weight Ratio (g/g) Males Females 0. 261 0. 276 0.314 0. 332 0.360 0.379 0.858 0.844 0.914 0. 684 0. 647 0. 682 0. 254 0.297 0. 288 0.333 0.440** 0.355 *. 0.843 0.918 0.891 0.655 0. 695 0. 563 0. 259 0.289 0.274 0. 267 0.284 0. 331 0.328* 0. 337 0. 337 0.332 0.775 0.851 0.739 0.927 0. 944 0. 611 0.574* 0. 608 0. 561 0.727* v* * Statistically significant difference at the 95 percent confidence level. ** Statistically significant difference at the 99 percent confidence level. TOXSTUDIES1171 'yarwjvioifv^ 1 S 3 1 - O 18 c T-I (1 ppm) T-II (10 ppm) O' T-III (100 ppm) Generation Fo FI F2 Fo FI F2 Fo FI F2 Fo FI TABLE XI TEST MATERIAL: Aroclor 1242 Three-Generation Reproduction Study - Albino Rats Mean Organ Weights and Ratios Parental Animals Organ: Brain Organ Weight (g) Males Females Organ/Body Weight Ratio (g/100 g) Males Females 2. 17 2.07 2. 01 1.90 1.93 1.74 0. 307 0. 334 0. 345 0. 509 0. 566 0. 557 2. 12 2. 07 2. 17 2. 21 , 2. 15 2. 12 . 2.08 1.95 1.96 1.91 1.99 Z. 05 1.93 1. 94 2. 01 1.77* 0. 303 0. 329 0. 328 0. 338 0. 346 0. 371 0.294 0. 302 0. 510 0. 637 0. 633 0. 542 0.577 0. 559 0. 540 0.464* * Statistically significant difference at the 95 percent confidence level. TOXSTUDIES1172 Q t% 'w w fo w ft JLS 3 1 * O 19 'y Generation c' Fo FI F2 . T-I (1 ppm) Fo FI F2 T-H (10 ppm) T-in (100 ppm) 2 ST * U1 Fo . FI F2 Fo FI TABLE XII TEST MATERIAL: Aroclor 1242 Three-Generation Reproduction Study - Albino Rats Mean Organ Weights and Ratios Parental Animals Organ: Adrenal Glands Organ Weight (g) Females Organ/liody Weight Ratio (g/100 g) Males ` F emales Organ/Brain Weight Ratio (g/g) Males Females 0.072 0.066 0. 071 0.065 0. 063 0. 061 0. 076 0.084* 0. 063 0. 095 0. 091 0. 080 0. 102 0. 077 0. 075 0. 093 0. 088 0. 088 0. 010 0. 010 0. 012 0. 009 0. 010 0. 009 0. Oil 0.013* 0. Oil 0. 025 0. 026 0. 026 0. 026 0.026 0.024 0. 024 0. 026 0.025 . 0.033 0. 032 0.035 0.030 0.031 0.028 0.034 0.039** 0. 030 0. 050 0. 047 0. 046 0. 052 0. 040 0. 038 0. 045 0.045 0. 045 0. 063 0. 069 0. 090 0. 078 0. 008 0.011 ' 0. 024 0. 020 0.030 0. 035 0. 039 0.044 ^Statistically significant difference at the 95 percent confidence level. ^Statistically significant difference at the 99 percent confidence level. TOXSTUDIES1173 B IO - T E S T JtaL&iaiMuiA., Site. Group Generation C Fo FI F2 T-I (1 ppm) Fo FI F2 T-n (10 ppm) T-III (100 ppm) c cO' * Fo , FI F2 Fo FI TEST MATERIAL: Aroclor 1242 Three-Generation Reproduction Study - Albino Rats Mean Organ Weights and Ratios Parental Animals Organ: Thyroid Gland Organ Weight (g) Males F emales 0. 035 0. 025 0. 034 0.028 0.025 0. 028 Organ/Body Weight Ratio (g/100 g) Males ` Females 0.0050 0.0040 0.0056 0.0075 0.0074 0.0089 Organ/Brain Weight Ratio (g/g) Males Females 0.016 0. 012 0. 016 0. 015 0. 013 0. 016 0. 036 0.034** 0. 038 0. 042 0.043** 0. 040 0. 031 0. 026 0. 030 0. 026 0.029* 0. 035 0.0051 0.0054** 0.0059 0.0081 0.0087 0.0095 0.0063* 0.0069** 0. 0070 0. 0067 0.0087 0.0099 , 0. 017 0.016** 0. 017 0. 019 0.020** 0. 019 0. 016 0. 014 0.015 0. 013 0. 018 o o * 0. 041 0.037* 0. 028 0.032* 0.0057 0.0p57* ' 0.0077 0.0082 0. 020 0.019** 0. 014 0. 018** * Statistically significant difference at the 95 percent confidence level. ** Statistically significant difference at the 99 percent confidence level. TOXSTUDIES1174 . -StiduAi/Ual B 1 O T E S T akvuit&iie4; Unc, 27 c. Histopathologic Findings Histopathologic findings among sacrificed animals are summarized in Tables XIV through XVI. Table XVII summarizes the histopathologic examination of the urinary bladder conducted on all animals sacrificed from the final generation. No tissue changes were observed microscopically which could be correlated with ingestion of test material. Findings among treated animals were essentially the same as those of control animals. 0365417 TOXSTUDIES1175 'fm yvwfo?- 1 S 3 1 " 0 1 9 Organ T rachea Lung Pancreas Kidney Adrenal gland EST MATERIAL: Aroclor 1242 Three-Generation Reproduction Study - Albino Rats Histopathologic Changes Control Group - Fo Generation Number of Animals Examined Findings Incidence 5 Males 5 Females Focal inflammation T racheitis Focal inflammation 3 1 1 5 Males 5 Females Focal inflammation Hyperemia Focal inflammation Hyperemia 2 5 2 5 5 Males Prominent islands of Lange rhans 1 5 Males 5 Females Focal inflammation Hyperemia Hyperemia 1 1 2 5 Females Hyperemia ' 3 Average Grade 0. 5 2. 0 1.0 1.0 2. 0 0. 5 1.0 1. 0 0. 5 0.5 1.0 1.0 fs> co 0 3 6 5 *il8 TOXSTUDIES1176 BI 0 -T EST Hoc. Organ Trachea Lung Liver TABLE XIV continued TEST MATERIAL: Aroclor 1242 Three-Generation Reproduction Study - Albino Rats Histopathologic Changes Test Group III - Fo Generation Number of Animals Examined 5 Males 5 Females , 5 Males 5 Females 5 Males 5 Females Findings Focal inflammation T racheitis Focal inflammation Focal inflammation Brown macrophages Hyperemia Focal inflammation Hyperemia Focal vacuolization Focal pericholangitis Hyperemia Incidence 2 2 3 1 1 1 1 -4 1 1 2 Average Grade 1.0 2.0 0.5 1.0 1.0 1.0 1.0 2. 0 0. 5 0. 5 1.0 TOXSTUDIES1177 i*4a l B I O - T E S T 2abo4aiMi&i, mP . Organ Kidney Adrenal gland TABLE XIV continued TEST MATERIAL: Aroclor 1242 Three-Generation Reproduction Study - Albino Rats Histopathologic Changes Test Group III - Fo Generation Number of Animals Examined 5 Males 5 Females Findings Focal inflammation Nephritis Hyperemia Nephritis Hyperemia ' Incidence 1 1 2 1 2 5 Males 5 Females Hyperemia Hyperemia 1 1 All other tissues and organs examined were normal. Grading System 0.5= Minimal 1.0 = Slight 2.0 = Mild 3. 0 = Moderate 4. 0 = Severe 5.0 = Extreme ' ^ Average Grade 0. 5 0. 5 0. 5 0. 5 .1.0 0. 5 0. 5 TOXSTUDIES1178 B IO - T E S T JedotauMiet, 9ftc. Orean Trachea Lung Liver Kidney Prostate TABLE XV TEST MATERIAL: Aroclor 1242 Three-Generation Reproduction Study - Albino Rats Histopathologic Changes Control Group - FI Generation Number of Animals Examined 5 Males 5 Females 5 Males 5 Females 5 Females 5 Males 5 Females 5 Males Findings Incidence Average Grade T rachcitis F ocal inflammation T racheitis 3 2. 0 3 ' 1.0 3 2. 0 Focal inflammation Focal inflammation Focal inflammation with abscess Hyperemia . 5 4 1 1 2. 0 1. 0 5. 0 2. 0 Hyperemia 1 0. 5 Nephrosis Nephritis Nephrosis 2. 1 1 0. 5 0. 5 0. 5 Focal inflammation 1 0. 5 T?*S9fO TOXSTUDIES1179 B 1O - T E S T TABLE XV continued TEST MATERIAL: Aroclor 1242 Three-Generation Reproduction Study - Albino Rats Histopathologic Changes Test Group III - FI Generation Organ Number of Animals Examined Findings Incidence Trachea 5 Males 5 Females T racheitis T racheitis Focal inflammation Lung . 5 Males 5 Females Focal inflammation Hyperemia Focal inflammation Focal inflammation with abscess Hyperemia . Liver 5 Males 5 Females Hyperemia Hyperemia Kidney 5 Males 5 Females Nephrosis Nephritis Nephrites Adrenal gland 5 Females Hyperemia All other tissues and organs examined were normal. Grading System 0.5 = Minimal 4.0 = Severe 1.0 = Slight 5.0 = Extreme 2. 0 = Mild 3 2 2 5 3 4 1 4 1 1 1 1 1 1 . Average Grade 2. 0 1. 0 0. 5 2. 0 1.0 0. 5 3. 0 2. 0 1.0 1.0 0. 5 0.5 0. 5 1.0 . TOXSTUDIES1180 7V<*9 0 <( fonvjVK'jo-f J. S 3 1 O I a Organ Trachea Lung TABLE XVI ' TEST MATERIAL: Aroclor 1242 Three-Generation Reproduction Study - Albino Rats Histopathologic Changes Control Group - F2 Generation Number of Animals Examined Findings Incidence 5 Males 5 Females 5 Males 5 Females Focal inflammation Inflammation Focal inflammation Inflammation Focal inflammation Inflammation with abscess Hyperemia Focal inflammation Inflammation with abscess Hyperemia 1 2 1 1 4 1 2 1 1 3 Average Grade 0. 5 3. 0 0. 5 1.0 1.0 3. 0 1.0 2. 0 4. 0 2. 0 T0XSTUDIES1181 B IO - T E S T la l& ia t& iio l, 2+tc. Orean Colon Liver Kidney TABLE XVI continued TEST MATERIAL: Aroclor 1242 Three-Generation Reproduction Study - Albino Rats Histopathologic Changes Control Group - FZ Generation Number of Animals Examined 5 Males 5 Females 5 Males 5 Females Findings Focal inflammation Hyperemia Focal inflammation NephrocalcinosLs Incidence 2 1 1 1 Average Grade 0. 5 0. 5 0. 5 0. 5 TOXSTUDIES1182 duAbual B I O - T EST Sue. TABLE XVI continued TEST MATERIAL: Aroclor 1242 Three-Generation Reproduction Study - Albino Rats Histopathologic Changes Test Group II - F2 Generation Organ Number of Animals Examined Findings T rachea 5 Males 5 Females Focal inflammation Focal inflammation Inflammation Lung 5 Males 5 Females Focal inflammation Focal hype'remia Hyperemia Focal inflammation Hyperemia Colon 5 Males Focal inflammation All other tissues and organs examined were normal. Grading System 0. 5 = Minimal 1.0= Slight 2. 0 = Mild 3.0 = Moderate 4.0= Severe 5.0 = Extreme Incidence 4 2 1 2 1 2 3 3 1 Average Grade 1.0 0.5 2.0 1.0 1.0 1.0 1.0 Z. 0 1.0 42V59C0 TOXSTUDIES1183 9?fc 59 CO v isa i-o ia Group C T-I (1 ppm) T-II (10 ppm) Sex M F M F M F TABLE XVII TEST MATERIAL: Aroclor 1242 Three-Generation Reproduction Study - Albino Rats Histopathologic Changes of the Urinary Bladder F2 Generation Number Examined Findings* Incidence 7 None 10 Focal epithelia hyperplasia 1 associated with focal cystitis (11) 7 Proteinaceous plug (30) 15 None ^ 5 Proteinaceous plug (55) Focal epithelial hyperplasia (69) 13 None 1 - 1 1 - * Individual animal numbers are shown in parenthesis. Grading System . 1.0 = Minimal or Trace 2.0 = Mild 3.0 = Moderate 4.0 = Severe 5.0 = Extreme Average Grade 1.0 . - 1.0 " TOXSTUDIES1184 $'nd*uUial BIO-TEST JahrtabvU&i, 9tic. 37 5. Reproductive Data Reproductive data for parental animals are presented in Table XVIII, The indices employed are defined below: Mating Index Number of Copulations51* ~ Number of Estrus Cycles Required X 100 Fertility Index Number of Pregnancies Number of Copulations X 100 Incidence of Pregnancy Number of Pregnancies Number of Females Mated X 100 Incidence of Parturition _ Number of Parturitions Number of Pregnancies X 100 The addition of 1 or 10 ppm Aroclor 1242 to the diet did not affect the reproductive performance of rats during the three-generation study. During the second generation (F2a and F2b litters) rats fed 100 ppm (T-III) had lowered mating indices and a reduced incidence of pregnancy. Because only two T-III females produced F2b litters it was not possible to continue this level through the third generation. * Only one copulation counted per estrus cycle. Five days equal one estrus cycle. . 036*427 TOXSTUDIES1185 9 7V<i`>tO wy 'wijn>w>py l S 3 1 ' O I 9 >,"N TABLE XVIII TEST MATERIAL: Aroclor 1242 Three-Generation Reproduction Study - Albino Rats Parental Animals Reproductive Performance Mating Index Group Litte r F raction Percent Incidence of Pregnancy F raction Percent Fertility Index F raction Percent Incidence of Parturition Fraction Percent C Fla 17/25 68.0 16/16 100. 0 16/17 94. 1 15/16 93.8 Fib 15/20 75. 0 15/15 100. 0 15/15 100. 0 15/15 100. 0 F2a F2b 16/37 14/26 43.2 53.8 14/16 14/14 87. 5 100.0 14/16 87. 5 14/14^ 100. 0 14/14 14/14 100.0 100.0 F3a F3b 15/66 12/16 22.7 75. 0 13/16 12/12 81.2 100. 0 13/15 12/12 86.7 100. 0 11/13 11/12 84.6 91.7 T-I Fla (1 ppm) Fib 16/37 15/21 43.2 71.4 16/16 15/15 100. 0 100.0 16/16 15/16 100. 0 93.8 16/16 15/15 100.0 100.0 F2a F2b F3a F3b 15/27 15/18 13/77 9/23 55. 6 83. 3 16.9 39.1 15/16 14/15 9/16 8/9 93. 8 93. 3 56.2 88. 8 15/15 14/15 9/13 8/9 100. 0 93. 3 '69. 2 88. 9 15/15 14/14 9/9 8/8 100. 0 100.0 100.0 100. 0 Gestation Time (Days) Mean Range 22 19-22 22 21-22 21 20-22 22 21-23 22 21-22 22 21-22 22 21-23 22 21-24 22 20-22 22 21-23 22 21-22 21 20-22 T0XSTUDIES1186 TABLE XVIII continued TEST MATERIAL: Aroclor 1242 Three-Generation Reproduction Study - Albino Rats Parental Animals Reproductive Performance Group Mating Index Litter Fraction Percent Incidence of Pregnancy Fraction Percent Fertility Index Fraction Percent incidence of Parturition Fraction Percent Gestation Time (Days) Mean Range T -II Fla 16/43 (10 ppm) Fib 16/24 37.2 66.7 16/16 16/16 100.0 100.0 16/16 16/16 100.0 100.0 15/16 16/16 93.8 100.0 22 22 21-23 21-23 F2a 13/53 F2b 13/24 24. 5 54.2 12/16 12/12 75. 0 100.0 12/13 12/13 92.3 92. 3 12/12 12/12 100.0 100.0 22 22 21-22 21-22 F3a 16/43 F3b 16/28 T-III Fla 15/43 (100 ppm) Fib 15/24 F2a 11/78 F2b 2/37 37. 2 57. 1 34.9 62. 5 14.1 5.4 16/16 15/16 15/16 15/15 7/16 2/7 100.0 93. 8 93. 8 100. 0 43.8 28. 6 16/16 15/16 15/15 15/15 7/11 2/2 100.0 93.8 100. 0 100. 0 63. 6 100. 0 15/16 15/15 14/15 15/15 7/7 2/2 93.8 100.0 93.3 100. 0 ' 22 22 22 22 100.0 100.0 21 22 21-22 22-23 21-23 21-23 - o u 9<9 > rv sC B I O - T E S T lobo-iaiiyiiei; 9nc. w T0XSTUDIES1187 ^nJuii'ual B I O - T E 5 T Jahvui&vUei, Phg. 40 B. Progeny 1. Population Data Population data for all litters are presented in Tables XIX and XX. The number of live pups born per litter and the number of pups weaned per litter were not affected by feeding 1 , 10 or 100 ppm Aroclor 1242. All pups delivered appeared outwardly normal and dis played normal growth through the lactation period. At weaning, an exam ination of each pup for external abnormalities was conducted and all pups were judged to be free of gross external abnormalities. 0365430 TOXSTUDIES1188 1 S 3 1 - O I 9 V*3*,tf'T V 'y Litter Fla Fib F2a F2b F3a F3b Fla Fib F2a F2b F3a F3b TABLE XIX TEST MATERIAL: Aroclorl242 Three-Generation Reproduction Study - Albino Rats Population Data Progeny Number of Pups Delivered Total Mean/Litter Number of Pups Stillborn T otal Mean /Litter Number of Pups Cannibalized Total Mean/Littei: Number of Viable Pups Born Total Mcan/Litter 178 191 147 164 102 109 11.9 12.7 10. 5 11.7 9.3 9.9 13 0.9 12 0. 8 0 1 7 0. 5 0 0 - '0 10 0.9 15 1.4 0 1 0. 1 _ _ 0. 1 165 178 140 164 92 93 11.0 11.9 10. 0 11. 7 8.4 8.5 ' 184 208 158 142 86 80 11.5 13. 9 10. 5 10. 1 9.5 10. 0 9 0. 6 11 0. 7 1 4 3 0.2 2 0.1 1 0 11 1.2 , '0 7 0. 9 4 0. 1 0. 3 0. 1 - 0. 5 174 193 154 140 75 71 10. 9 12.9 10.3 10. 0 8. 3 8.9 TE<>99fO TOXSTUDIES1189 tu b u h ia l B I O - T E S T J a & Q id a ile l, Hue. Group Litter T -II (10 ppm) Fla Fib F2a FZb F3a F3b T-III . Fla (100 ppm) Fib F2a F2b TABLE XIX continued TEST MATERIAL: Aroclor 1242 Three-Generation Reproduction Study - Albino Rats Population Data Progeny Number of Pups Delivered Total Mean/Litter Number of Pups Stillborn Total Mean/Littcr Number of Pups Cannibalized Total Mean/Litter Number of Viable Pups Born Total Mean/Litter 176 207 126 138 149 171 11.7 12.9 10. 5 11.5 9.9 11.4 3 4 8 5 17 8 0. 2 0. 2 0.7 0.4 1. 1 0. 5 0 0 0 0 _ - 5 0. 3 3 0. 2 173 203 118 133 127 160 11.5 12. 7 9. 8 11.1 8. 5 10. 7 168 203 12.0 13. 5 8. 3 0. 6 0. 2 . 4 0.3 ' 0 156 200 11. 1 13. 3 53 7. 6 6 20 10.0 2 0. 9 1.0 2 0. 3 2 1. 0 45 6.4 16 8. 0 TOXSTUDIES1190 BIO -TEST Jc TABLE XX TEST MATERIAL: Aroclor 1242 Three-Generation Reproduction Study - Albino Rats Population Data Progeny jtoud Litter Number of Pups at Lactation Day 1 Total Mean/Litter Number of Pups at Lactation Day 5 T otal Mean/Litter Number of Pups Retained at Lactation Day 5 Total Mcan/Littcr Male Number of "Weanlings F emale Total Mean/ Litter Fla Fib 146 176 9.1 11.7 70 117 4.7 7. 8 66 103 4.4 6.9 34 28 31 35 62 4. 1 66 4.4 F2a F2b 132 163 9.4 11. 6 90 135 6. 4 9.6 78 115 5.6 8. 2 30 34 64 4. 6 39 46 85 6.1 F3a F3b 90 91 8. 2 8. 3 69 6. 3 65 5.9 26 25 51 4.6 79 7. 2 75 6.8 16 26 42 3.8 r-i Fla ;i ppm) Fib' W F2a F2b 176 175# 152 133 11.0 12. 5 10. 1 9.5 103 148 107 100 6.4 91 5. 7 36 53 89 5.6 10. 6 IZ0 8. 6 46 40 86 6.1 7. 1 7. 1 99 6. 6 . 90 . 6-4 37 39 76 5. 1 37 30 67 4.8 F3a F3b 69 66 7. 7 8. 2 46 5. 1 39 4. 3 12 13 25 2.8 46 5.8 45 5. 6 12 11 23 2. 9 v One female died on Day 1 of lactation. Pups from this litter were sacrificed and are not included in the totals and means. T0XSTUDIES1191 1S 31-O I9 0U(f TABLE XX continued TEST MATERIAL: Aroclor 1242 Three-Generation Reproduction Study - Albino Rats Population Data Progeny Stovlp________Litter Number of Pups at Lactation Day 1 Total Mean/Litter Number of Pups at Lactation Day 5 Total Mean/Litter Number of Pups Retained at Lactation Day 5 Total Mean/Litter Number of Weanlings Male Female Total Mean/Litter T-H (10 ppm) Fla Fib 170 202 . 11.3 12. 6 133 163 8.9 10. 2 118 7.9 134 8.4 59 49 46 60 108 106 7. 2 6.6 F2a F2b 109 132 9.1 11.0 96 8. 0 ,90 7. 5 38 39 115 9.6 103 8.6 37 37 77 6.4 74 6. 2 F3a F3b 126 152 3.4 10. 1 114 7.6 105 7. 0 44 60 104 6.9 119 7.9 102 6.8 51 46 87 5.8 r-in (100 ppm) Fla Fib 149 196 10. 6 13. 1 133 9.5 122 8. 7 44 51 133 8.9 '' 111 7.4 31 28 95 6.8 59 3. 9 F2a F2b 37 15 5.3 7.5 36 5. 1 15 7.5 36 13 . 5. 1 6.5 14 21 56 35 5.0 11 5.5 <7 s* T0XSTUDIES1192 frndtuUial B I O - T E S T J.aimrUoAiei-, 9nc. 45 2. Survival Data Survival data for progeny are presented in Table XXI, Live birth, 24 hour, five day and lactation indices were normal for all groups. The indices employed are defined below. Live Birth Index Number of Viable Pups Born Total Number of Pups Born 24-Hour Survival Index Number of Pups Viable at Lactation Day 1 x 100 Number of Viable Pups Born 5-Day Survival Index Number of Pups Viable at Lactation Day 5 x jqq . Number of Viable Pups Born Lactation Ind^x - Number of Pups Viable at Lactation Day 21 x jqq Number of Pups Viable at Lactation Day 5 0365*35 TOXSTUDIES1193 ; jIndu&btial BIO-TEST Hc. 46 ( TABLE XXI TEST MATERIAL: Aroclor 1242 Three-Generation Reproduction Study - Albino Rats Survival Indices Progeny Group C T-I (1 ppm) Live Birth Litter Index Fla Fib F2a F2b F3a F3b 92. 7 93.4 95.2 100. 0 90. 2 85. 3 Fla Fib F2a F2b F3a F3b 94.6 92. 3 97.5 95.5 87. 2 88. 8 24-Hour 5 -Day Survival Index Survival Index Lactation Index 88. 5 98.9 94. 3 99. 3 97. 8 97. 8 42.4 65. 7 64. 3 82. 3 75. 0 85.0 93.9 64.1 81.2 73.9 78. 5 56.0 98. 3 97. 2 98. 7 95.0 92. 0 93. 0 59.2 82. 2 69. 5 71.4 61.3 63.4 97.8 71.7 . 76.8 74.4 64.1 51.1 T -II (10 ppm) Fla Fib F2a F2b F3a F3b 98. 3 98. 1 93.7 96.3 85.2 94.1 98. 3 99.5 92.4 99.2 99.2 95.0 76. 9 80.3 81.4 86.4 82. 7 74. 4 91.5 79. 1 85.6 71.8 99.0 85. 3 T -III (I 00 ppm) Fla Fib F2a F2b 92.9 98. 5 84.9 80,8 95.5 98. 0 82. 2 93.8 85.3 66.5 80.0 93. 8 77.9 53.2 97.2 84. 6 t 0365*36 TOXSTUDIES1194 ytiduAkUal BIO TEST hyiabyueS, Hkc. 47 3, Bodv Weight Data . Body weight data for weanlings are presented in Table XXII. There are no significant differences between test and con- trol weanling body weights. v TOXSTUDIES1195 Jltulu&Uial B I O - T E S T Jaho'iaU'Uei., 9hc. 48 Group C T-I (1 ppm) T -II (10 ppm) T -III (100 ppm) TABLE XXII TEST MATERIAL; Ax odor 1242 Three-Generation Reproduction Study - Albino Rats Progeny Weanling Body Weight Data Litter Mean Body W eight (grams) Male Female Fla Fib F2a F2b F3a F3b Fla Fib F2a F2b F3a F3b Fla Fib F2a F 2b F3a F3b Fla Fib F2a F2b 44 47 46 41 45 42 46 46 47 45 38 35 49 52 48 41 49 39 ' 45 42 55 38 44 43 43 39 44 42 ' 46 45 - 44 40 38 39 47 47 47 45 46 43 . 43 42 48 37 ( 03640# T0XSTUDIES1196 BIO-T E S T $nc. 49 4, Pathologic Studies At autopsy of the F3b weanlings no gross or microscopic tissue changes were present which could be associated with the ingestion of Aroclor 1242 . ( C 0365439 TOXSTUDIES1197