Document 0qz6RKqBLLq643vNVbvr3wBXk

A1* .^JDALL JON'/ ESQ. iARLES H. McC/A, SR., ESQ. NIK SKRINJARir ESQ. JONES f JONES,-"LOSE & BROWN, CHARTERED t 300 -South FoVth Street #300 "Las Vegas, 1^ 89101-6026 '^Telephone: ?02) 385-4202 y '' ATTORNEYS PLAINTIFF ' ... A UNITED STATES DISTRICT COURT DISTRICT OF NEVADA * NEVADA PQtfR COMPANY, a Nevada corporatio, *7 Plaintiff, OLj TO COMPANY, a foreign Ugon; GENERAL ELECTRIC Y1, a foreign corporation; NGHOUSE ELECTRIC ATION, a foreign tion; and DOES I through elusive,' 4 Defendants. CV-S-89-555-LDG-LRL OPPOSITION TO DEFENDANT'S JOINT MOTION FOR SUMMARY JUDGMENT AND JUDGMENT ON THE PLEADINGS AND REQUEST FOR ORAL ARGUMENT EXHIBIT 9 VOLUME II R EPO R T TO MONSANTO COMPANY T W O-Y EAR CHRONIC ORAL TOXICITY WITH AROCLOR 1260 IN. ALBINO RATS NOVEMBER 12, 1971 IBT NO. B7298 L Introduction At the r e q u e s t of Monsanto Company, a tw o - y e a r chronic o r a l toxicity study was conducted using albino ra ts :o determ ine the potential toxicity of A r o c lo r 1260, Lot No. AK-3. The following re p o rt p r e s e n t s the results of this investigation. CO^ 000082 3 In Tc ST IL Sum m ary 2 Food consumption and body weight gains were not altered at any le v e l of A r o c l o r 1260 fed. The n u m b e r of a n i m a l s dying and the time at which the deaths occurred did not vary among the test and control groups. Hematological and clinical blood chem istry studies conducted after 1260. Urine analyses failed to reveal any differences between test and control animals. At s a c r i f i c e a f t e r 3, 6, or 12 m o n th s on t e s t o r g a n w e ig h ts , o r g a n to body weigh: and organ to b r a n weight ratios disclosed several randomly occurring intergroup differences. The lack of any consistent dose related re sponse and the absence of any deleterious histopathologic change confirm that these d ifferen c es w ere not related to the ingestion of A roclor 1260. At the final sacrifice after 24 months on test, the liver weights and liv e r to body weight or b rain weight ratios w ere significantly elevated in the rare f r o m the T-UI group. Histologic e x a m i n a t i o n of the liv e rs from the T-UI group revealed several animals with vacuolar change. This le sio n is m o rpho lo g ically indicative of fatty deg en eratio n . Specific fat stains c o n f i n e d the p resen ce of fat in these vacuoles. Focal hypertrophy and focal h y p e rp la sia were also found in the liv e r s from animals fed Aroclo 1260. A0H 003721 263 000083 % o Hyperplasia of the urinary bladder was found in an animal from the control group but not in any of the te st anim als. T h e i n c i d e n c e s a n d types of aU t u m o r s w e r e about the s a m e ip. a l l groups, including the control group, and are considered normal .... of this *ge. Respectfully submitted, INDLSRIAL 3 IO -T Z S T LA3CRATCRIZS, INC Report p re p a re d by: 1L\ P h i l i p 3 m i t B . S. Assistant Toxicologist F-at T oxi ci ty Report approved by: Jkrn^s 3. Plan* Sector Group Leader R-at T o x i c i t y <TJ ' / / / , . y } * . P a u l L. Wright, ? h . D. Section Head, Toxicology L. ^epliqg/^7^?h. Manager, Toxicology N ovem ber 12, 1971 lam :sjn *0" 03722 264 000084 *1 I m. Procedure A. E x p e r i m e n t a l Animals The an im als employed in the test were C harles River strain-* albino r a t s . F o u r hundred ra ts (200 m a le s and 200 fe m ale s) were selected for the experim ent, ear-punched with the animal num ber aeeigned and housed individually in standard w ire-bottom ed steel rat cages. Each cage bore a color-coded card identifying the rat with r e spect to p ro ject num ber, dose level assignment, individual animal number and sex. B . Organization of Groups A s t r u c t u r a l outline of the ex p erim en t is given in Table L O TABLE I TEST MATERIAL: A ro c lo r 1260 Tw o-Year Chronic Oral Toxicity Study - Albino Rats Outline of E x p e rim e n t Crouo N u m b e r of Animals Male F errale Dietary Level (oor.) Control 50 50 None Administered T - I 50 50 1 T-II 50 50 10 T-I1I 50 50 100 Charles River Breeding Laboratories, North Wilmington, Mass. ACM CC3722 265 000085 C* D ie ts . Feed ing and Food Cor. sumo tion All d i e t s w e r e p r e p a r e d Li the c e n t r a l d ie t r e e m of i s laboratory. The basic ration from which all diets were constructed was a standard, pulverized stock rat ration*. The diet for any given t e s t group was p r e p a r e d by blending the calculated amount of A ro c lo r 1260 with a p r e - w e i g h e d p o r ti o n of the stock r a t i o n Ln a H o b a r t m i x e r . F resh diets were prepared each week and every rat was offered an am ount of food sufficient for ad libitum feeding. Food consumption was recorded for five r a t s of each sex in every group weekly for the first three months and monthly for the next nine months. Thereafter, periodic spot checks were made. D. Bodv Weights and Weight Cains In it ially, the. body weight of each r a t in e v e r y g ro u p was d e t e r mined and recorded. Thereafter, individual weighings were made weekly f o r the f i r s t 13 w eek s and m ont hly t h e r e a f t e r u n ti l the co nclu si on of the i n v e s ti g a ti o n .___________________ __________________________ ________________ E. Mortality and Reactions * Checks for mortality and abnormal behavioral reactions were .made daily throughout the investigation. 7-T* F . H e m ato l o g ic Studies and Urine A naly ses Blood studies, including d e te r m in a tio n s of hemoglobin c o n c e n t r a tions, hem otocrit value, erythrocyte count and both total and differential P u r i n a Rat Chow, Ralston Purina C o ., St. Louis, Mo. AC* CQ37 266 00008G o leukocyte counts, were conducted upon five m a le s and five fem ales from b o t h the c o n t r o l and 100 pp m gro u p s . U rina a n a ly s e s for the pre s e n c e of glucose, albumin, m i c r o scopic e l e m e n t s , and d e te r m in a tio n s of pH and specific gravity w ere c o n ducted upon urine sam ples at the same time intervals and from the sam e an im a ls as those employed for the blood studies. T h e s e s t u d i e s w e r e conducted a f t e r 3, 6, 9, 12, 18, and 24 m o n th s of feeding. C. Clinical Blood Chemistry Studies D e t e r m i n a t i o n s of blood u r e a n i t r o g e n c o n c e n t r a t i o n (BUN)# e r u m alkaline phosphatase activity (SAP), fasted blood glucose .concen tration and s eru m glutam ic-pyruvic tran sam in ase activity (SGPT) were c o n d u c t e d a f t e r 3, 6, 9, 12, 18, and 24 m o n th s of feeding upon the s a m e ra ts as w ere the routine blood studies. H. Pathologic Studies When an animal succumbed during the test period, a gross autopsy was p e rfo rm e d . In those cases when p o s tm o rte m autolysis was not a d vanced, rep resen tativ e tissues were taken and p re serv e d in ten percent form alin solution for possible future histopathologic study. A f t e r 3, 6, and 12 m onth s of te st in g , five a n i m a l s of each s e x from each group were sacrificed and subjected to complete gross pathologic examinations. ADM 0C372 267 000087 Absolute organ weights w ere r e c o r d e d ar.d organ to body weight and organ to brain weight ratios computed. The following organs were included: liver, kidneys, spleen, gcr.ads, h e a rt and brain. These data w e r e then subjected to s ta t i s t i c a l analysis. An Analysis of V ariance was conducted first and significant effects disclosed by that treatment were fu rth er studied by " f '- t e s t s . Complete m icroscopic examinations were conducted upon the t i s s u e s and o r g a n s f r o m a ll c o n t r o l and 100 p p m gro up r a t s s a c r i f i c e d a f t e r 3, 6, o r 12 m o n th s of t e s t i n g . Ths following t i s s u e s and o r g a n s w e r e included in the examinations: heart, trachea, lungs, liver, pancreas, esophagus, stomach, small intestine (duodenum, jejunum and ileum), caecum, colon, spleen, lymph nodes, kidneys, urinary bladder, gonads, prostate gland, seminal vesicles, uterus, pituitary gland, adrenal glands, salivary glands, thyroid gland, parathyroid glands, skeletal muscle, sternum, bone, peripheral nerve, spinal cord and brain (cerebrum, cerebellum and pons). The aforementioned tissues were prepared and stained with Hematoxylin-Eosin stain. I" * A fte r two years of feeding, all surviving anim als were sacrificed and subjected to complete gross pathologic examination. The absolute weights of the liver, kidneys, spleen, gonads, heart and brain were recorded immediately after sacrifice. ADH 0 0 3 7 2 268 000083 O M icroscopic examinations were conducted upon tissues and organs from selected animals dying during the experiment and upon all anim als from the final sacrifice. T issues and organs examined were the same as those previousl * c mentioned. L Tumor Incidence and Classification A tabulation of the incidence of tu m o r formation in each group w a s m a d e at the conclusion of the investigation. In addition, t u m o r data for individual animals including location, weight, sire and pathologic classification were recorded. ACM C C 3 7 2 7 269 000089 XV. R e s u l t a A* Bo dv Weights and Weight Gains The m ale and female body weight data are assem bled in Table II. The d a t a m this t a b le a l s o in cl ude the 3, 12 and 24 m onth to ta l weight I9 gains. T h e r e w e re no significant effects due to ingestion of the compound A O CC 3 270 oooosn k )ietary Level of A ro c lo r 1260 |pprr>> Sex Control M F K-J> | M F 10 M F 100 M F OQ T A B L E II T E ST MATERIAL: A ro c lo r 1260 Tw o-Year Chronic Oral Toxicity Study - Albino Rats Body Weight and Total Weight Gain Data Sum m ary of Mean Values Body Weight 90-Day (grama) Total Weight Week: Gain 0 1 2 3 4 5 6 7 8 9 10 11 12 1) ( g r a m a / r a l j 13) 203 245 313 362 373 391 409 424 440 440 449 467 474 136 178 190 219 244 254 259 265 270 276 281 278 290 288 34 1 152 133 192 250 322 361 374 394 4 16 428 448 450 474 479 490 136 167 189 218 24 1 252 256 261 268 27 ) 277 281 290 288 357 152 133 204 245 317 368 382 400 417 436 448 461 467 475 481 136 174 190 217 242 254 260 263 268 274 281 279 285 288 348 152 13) 208 247 319 363 375 394 425 437 451 460 479 486 500 136 173 188 222 240 253 258 262 265 267 270 275 275 2U0 367 14 4 o 3 or o .i t I 60000 Dietary Level of A ro c lo r 1260 (ppm) Sex Control M F 1 ro NJ 10 M F M F 100 M F O T A B L E II continued T E S T M ATERIA L: A ro c lo r 1260 Tw o-Y ear Chronic Oral Toxicity Study - Albino Rata Body Weight and Total Weight Gain DaU Summ ary of Mean Values Body Weight (yrama) Month: 45 6 7 8 9 10 1 1 498 533 565 593 622 642 667 688 297 310 322 333 345 359 374 383 516 547 578 600 628 643 668 688 298 307 314 321 333 340 353 367 496 526 551 581 60 6 633 660 685 291 306 314 3 i 1 340 358 370 381 522 555 580 607 633 650 681 707 289 302 307 314 326 333 342 347 O 12-Month Total Weight Ga in 12 ( r a m s / r a t ) 701 568 390 254 699 4 379 566 243 709 576 388 252 717 584 36 3 227 000092 > o 2 O o 1>J u-a Dietary Level of A r o c lo r 1260 (ppm) Sex Control M F 1M F 10 M F 100 M F O o r> to x -J o o ui OO T A l i L E II continued T E S T MATERIAL: A ro c lo r 1260 Two-Year Chronic Oral Toxicity Study - Albino R aft Body Weight and Total Weight Gain Data Sum m ary of Mean Value Body Weight 24-Month (grama) Total Weight Month: Gain 13 14 IS 16 17 18 19 20 21 22 23 24 (g r a m a / r a t ) 728 746 759 768 769 767 760 758 739 * 726 687 642 426 444 466 481 500 515 533 547 551 550 564 551 721 740 760 768 776 774 762 753 733 718 715 654 390 427 466 433 445 476 481 493 514 5 30 484 525 509 4 15 52 1 385 728 745 776 761 768 7H0 774 761 735 715 683 645 416 434 482 449 473 508 519 5 30 5 36 542 536 584 746 760 772 775 774 769 760 750 733 714 711 64 1 380 386 417 414 435 453 471 481 500 507 485 537 5 12 448 508 40 1 I I 13 B. Food Consumption Tilt data obtained from the food consumption m easurem ents c on duc te d d u r i n g the f i r s t 12 m o n t h s of feeding a r e s u m m a r i z e d in T a b l e III. T h e se data revealed no significant difference between test and control rats. The periodic checks conducted during the rem ainder of the investigation also revealed no outstanding differences between te st and control animals. ( -e ACP CC3 0 0 274 vJ O o T A B L E III T E ST MATERIAL: A ro c lo r 1260 Two-Year Chronic Oral Toxicity Study - Albino Rata Food Consumption Data Sum m ary of Mean Values 1 Food Consumption letary Level of (gram a/rat/aeven days) Average A r o c l o r 1260 Week: Conuump (ppm) Sex 1 2 3 4 5 6 7 8 9 10 11 12 11 ( g r a m s / r a t / s c Control M 94 160 175 161 117 137 225 197 204 188 191 194 196 F 7 1 140 144 107 104 111 208 154 165 151 146 150 151 174 140 1 M 118 154 151 169 201 116 200 198 201 191 189 187 194 F 95 118 97 119 160 107 162 119 165 149 150 141 147 175 116 10 M 106 146 144 142 190 116 196 214 214 192 191 199 200 F 94 124 128 124 179 90 190 167 155 151 150 49 152 175 141 100 O O M 121 150 156 117 187 124 121 190 196 186 184 199 189 F 89 91 121 151 151 101 191 116 140 144 144 147 H 6 161 1 15 ACM 0037 r- o ietary Level of A r o c lo r 1260 (ppm) Sex 4 Cont rol M 196 F 148 l M 196 F 148 10 M 186 F 142 100 M 179 F 156 TA B LE III continued TE ST MATERIAL: A ro c lo r 1260 Two-Year Chronic Oral Toxicity Study - Albino Rata Food Consumption Data Summ ary of Mean Values Food Consumption (gram s/rat/seven days) Average Food Month: Consumption 5 6 7 8 9 10 11 12 - ( g r-a.m....s. / r a t /--s --c v--c--n---(-l--av1s- )1 189 168 147 163 184 179 170 187 154 136 121 138 152 160 141 127 176 142 200 189 192 179 181 182 172 184 163 141 163 160 153 157 143 ' 132 186 151 191 188 193 197 197 168 164 196 138 145 144 144 151 136 139 141 186 142 170 180 188 186 166 181 188 180 127 147 144 134 132 162 149 139 * 180 143 l vo X a oVP VP 00 276 C. M o r ta lit y and Reactions A f r t q u t n c y distribution of natural deaths o c cu rrin g during the i n v e s t i g a t i o n a p p e a r s in T a b le IV. No untow ard b eh av io ral re a c tio n s w e r e noted among *ny of the animals employed in the investigation. AO* CC37 00 277 O Dietary Level (ppm) Sex Cont rol M F 1M F 10 M F 100 M F T A B L E IV T E S T M A T ER IA L ; A r o c lo r 1260 Two-Year Chronic Oral Toxicity Study - Albino Rata Mortality Data Frequency Distribution of Deaths Frequency Distribution of Deaths Months; - 1-3 4-6 7-9 10-12 13-15 16-18 19-21 000 2 17 6 000 1 3 5 7 00 1 1 4 7 5 000 2 3 3 6 1 10 2 4 7 8 000 3 24 5 10 0 2 2 8 6 0 22 0 35 7 22-24 9 3 8 5 6 3 9 4 Total Dead Number Tested* 25/35 19/35 26/35 19/35 29/35 * 17/35 29/35 23/35 a Does not include the live anim ala of each aex fro m e v er y group sa c rif ic e d for in te r im pathologic stud ic a . nx OC 3 73 WI W I %* 4 13 D. H em atologic and Clinical Blood1C h em istry Studies M ean m ale and fem ale hem atologic and clin ical blood c h e m istry d a t a a r e s u m m a r i z e d in T a b le s V th ro u g h DC. Values for all p aram eters investigated w ere within the norm al * I range for the albino rat. No significant differences between test and control values w ere observed. ACM C C 3 1 3 - 00' 279 i B I O T HS T O OO Dietary Level (ppm) Control Sex 3 M 14.0 F 9. 1 TABLE V TEST M ATERIAL: A roclor 1260 Tw o-Y ear C hronic O ral Toxicity Study - Albino Rata Hematologic Data Sum m ary of Mean Valuea Total Leukocyte Count fthou aand a/m m Month: 6 9 12 18 24 Erythrocyte Count (mill tone/mm*) Month: 36 9 12 18 IS. 4 9.4 10.1 6.7 II.1 7.6 9.9 8.8 11.2 7.6 8. 1 8 .2 8.7 8 .8 7 .6 7.5 7.7 7.3 7.0 7.7 24 6.5 6. B M 10.8 13.1 7.9 12.8 8.6 8.4 100 F 13.2 8.2 7.9 6.5 7.8 7.4 7.5 8.1 8.0 7.6 8.0 7.6 7.3 7.5 7.3 7.5 7.4 6.4 08Z JO >o Oo .I It I *1 Dietary Level (ppm) Sex 3 M 17. 1 Control F 16.9 O TABLE VI TEST M ATERIAL; A ro d o r 1260 Tw o-Y ear Chronic O ral Toxicity Study - Albino Rate Hematologic Data Sum m ary of Mean Values Hemoglobin Concentration (g/100 ml) Month; 6 9 12 18 24 H em atocrit Value <%> Month; 3 6 9 12 17.7 17.4 15.7 17.6 . 14. 1 16.7 16. 1 13.6 16.4 14.6 51 49 48 50 50 47 43 46 18 41 42 M I6l 1 17.2* 14.0 15.3 15.9 14.5 100 F 16.3 17.0 13.7 15.6 15.3 12.9 49 47 43 43 41 48 47 41 44 39 24 35 38 38 a 32 r ACM 0C3 I~tJi o Dietary Level (ppm) Sex 3 Control M 82.0 F 83.0 TABLE VII T E ST M ATERIAL: A ro c lo r 1260 Tw o-Y ear Chronic Oral Toxicity Study - Albino Rata " Hematologic Data Summ ary of Mean Values 6 86.6 86.6 Differential Leukocyte Count (Number of Cells per Hundred! L ym phocytes Neutrophils Month: Month: 9 12 18 24 3 6 9 12 18 80.4 78.8 79.0 65.4 16.0 11.4 18.6 20.6 20.0 86.8 89.0 70.2 61.4 15.4 11.6 11.8 10.0 29.2 24 32. 2 36.0 M 85.8 85.4 83.4 72.4 7 3 . 8 66 6 100 F 92.0 89.6 78.8 82.2 80.6 65.6 12.6 7.6 13.8 10.0 15.8 19.4 22.6 17.0 24.6 17.4 31.6 34.2 00 D OOiiJ x- ft Dietary Level Cj>pm) Sex 3 Control M 0.6 F 0.6 M 0.0 100 F 0.0 TABLE VI! continued TEST M ATERIAL; A ro c lo r 1260 Tw o-Y ear Chronic O ral Toxicity Study - Albino Rata Hem atologic Data Sum m ary of Mean Values Differential Leukocyte Count (Number of Celia per Hundred) M onocytes Eosinophlla Month: Month: 6 9 12 18 24 36 9 12 0 .0 0 .0 0.4 0. 2 1.2 1.4 2.0 1.0 0. 2 0 .0 0.0 0.0 0.2 1.0 1.0 1.8 1.4 0. 2 18 0 .8 0.6 24 1.6 1.6 0.0 0.0 0.0 0.4 0.8 0.0 0.0 0.0 0.4 0.2 1 .4 0 . 8 0. B 1 .0 1. 2 1.00.4 0.4 0 .8 0.8 1.6 0 .0 k> o2 Oo m 0 ( 283 Q I O TABLE! VII continued TEST M ATERIAL; A r o d o r 1260 Tw o-Y ear Chronic O ral Toxicity Study - Albino Rate Hem atologic Data Sum m ary of Mean Values Dietary Level (ppm) Sex Differential Leukocyte Count (Number of Celle per Hundred) Basophils Month: 6 9 12 18 M 0.0 0.0 0.0 0.0 0.0 Control F 0.0 0.0 0. 0 0.0 0 .0 M 0.2 0.0 o e 0.0 0.0 100 F 0.0 0.0 0.0 0.0 0.0 o 24 0.0 0.0 0.0 0.0 284 O * o X o o m O I fl Dietary Level Ippm) Sex 3 Control M 38 F 33 M 33 100 F 20 O TABLE VIII TEST MATERIAL: A roclor 1260 Two*Year Chronic O ral Toxicity Study - Albino Rata C linical Blood C h em istry Data Summary of Mean Values Serum Glutamic-Pyruvic T ransam inase Activity (Dade Units) Month: 6 9 12 IB 24 36 Blood U rea Nitrogen Concentration (m g s %) Month: 9 12 18 24 48 45 45 42 36 39 38 42 32 30 17 15 12 10 14 15 14 16 15 9 13 11 39 42 43 31 36 37 50 40 32 33 14 15 17 16 16 15 15 15 14 13 12 14 O <=> o* toLoon X o r Ut a*. c /i* ' D ietary Level (ppm) Sex 3 Control M 128 F 130 1 100 M 118 F 117 TA B LE IX TEST MATERLALx A roclor 1260 Tw o-Y ear Chronic O ral Toxicity Study - Albino Rata C linical Blood C hem istry Data Sum m ary of Mean Values F asted Blood Glucose Concentration fmga%) Month; 6 0 12 18 24 Serum Alkaline Phosphatase Activity (K in g - A r m s tro n g U n iti>) Month: 3 6 9 12 18 24 123 89 118 U S 88 138 99 92 134 109 28.6 18.9 24.7 17.8 21.5 10.6 20. 3 11.9 16.2 16.8 22. 3 20. 2 f. o C 118 116 135 146 98 116 1(5 138 146 94 37.8 12.0 21.8 21.9 12. 1 7 .6 23.9 11.7 11.9 7.8 16. 1 12.4 O cU^t AOM GOJ Urina Analyses M e i n m a l a And f e m a le u r i n e a n a l y s e s d ata a r e s u m m a r i s e d in T a b le s X th ro u g h XXL Determinations for glucose concentration were negative for all % ra ta a t every in terv al of exam ination. Ko`d ifferen ces w ere noted betw een the urine from control and te st anim als at the intervals of examination. ACM 003 00 287 l iu U B I n T 5 T Dietary Level PPTM> Sex 3 Control Mn Fn Mn 100 Fn oo TABLE X T E S T M ATERIAL: A ro clo r 1260 Tw o-Y ear Chronic O ral Toxicity Study - Albino Rata Urine Analyses Data Sum m ary of Mean Valuea Glucoae Month: 6 9 12 nnn nnn 18 24 nn nn Albumin Month: 3 6 9 12 18 24 n n I t 42 43 nnnnt 42 n n n nn n n n nn n t t t 4 1 43 n n n n 4 1 44 P^iCOO wov Key: n s negative t = tr a c e ; leaa than 30 m g / 100 m l u rin e + 1 = 30 to 100 m g / 100 m l u r i n e 4 2 = 100 to 300 m g / 100 m l u r in p * 3 = 300 to 600 m g / 100 m l u r in e 4 = m o r e than 500 m g / 100 m l u r in e * O Dietary Level (ppm) Sex a Control MN FN MN 100 FN U T A B L E XI TEST M ATERIAL; A ro clo r 1260 Tw o-Y ear Chronic O ral Toxicity Study - Albino Rate Urine Analyses Data Sum m ary of Mean Values M icroscopic Elements Month; 6 9 12 IB NNNN NNNt 24 1 t a6 pH Month; 9 12 7767 8 B7 8 N N N Nt N N N tt 7767 7 *8 6 7 O 18 24 76 66 66 66 Key; N= N o r m a l I s trace; less than 5 per low pow er field O i l = 5 to 10 p e r low p o w e r fie ld O ' i 2 s 10 to IS p e r low p o w e r fie ld * ' O - m o re than IS p e r low pow er field r L O nt t/l N Oi 289 A IO r Dietary Level (ppm) Control too O TAfaLE XII T E ST M ATERIAL: A roclor 1260 Tw o-Y ear Chronic O ral Toxicity Study - Albino Rats Urine Analyses Data Summ ary of Mean Values n u - , Sex 3 M 1.029 F 1.020 M 1.021 F . 1.016 6 1.022 1.019 1.025 1.023 Specific G ravity Month: 9 12 1.033 1.027 1.025 1.024 1.031 1.035 ` 1.030 1.029 18 1.042 1.032 1.043 1.028 24 1.051 1.039 1.039 1.045 * 290 O > o oo U> JmJ+Uual B ' O * T E S T c. 30 F . Pathologic Studies 1 T h r e e - M o n t h S a c r i f i c e a. Cross Pathologic Findings No outstanding differences w ere noted between te st and control ra ts upon gross pathological examination. b. Organ Weight and Organ to 3odv and Organ to Brain Weignt Ratio Data The results of the statistical analyses conducted on absolute organ weights organ to body w eight and organ to b ra in w eight r a t i o s a r e s u m m a r i z e d in T a b le s -XIII th ro u g h XVIII. Significant differences between a test group and the control group are designated by asterisk s following the test values. Organ weights organ to body weight and organ to b ra in weight ratios disclosed several randomly occurring intergroup difference: The lack of any consistent dose related resp o n se and the absence of any deleterious, histopathologic change confirm that these differences w ere not re la te d to the ingestion of A roclor 1260. AON C037<r 291 0 n r\ n / oo TA BLE XIII T E ST MATERIAL A ro c lo r 1260 Tw o-Y ear Chronic O ral Toxicity Study - Albino R ata Organ Weight and Ratio Data Three-Month Sacrifice Sum m ary of Mean Value* Organ; Liver Dietary Level of A ro clo r 1260 (ppm) O rgan Weight fg) Male* F em ale* Organ/Body Weight Ratio (g /1 0 0 g) M ales Fem ales O rgan/B rain Weight R atio (g/gl M ales Fem ale s Cont rol 21.4 13.0 4.13 4.16 10.3 6.82 l Vto> IP to too 20.4 24.044 29.544 13.2 12.2 14.5 3.90 4.48 5.7344 4.59 3.61 4.58 9.98 11.8 14.44# 6.86 6.38 7.72 Statistically significant difference at the 99 percent confidence level. >o 3 k ) OoU> -VJJ> o ytm ' it 1 cn m l/t o I Dietary Level of A ro c lo r 1260 (ppm| Control 1 10 100 o TA B LE XIV TEST M ATERIAL: A roclor 1260 T w o-Y ear C hronic O ral Toxicity Study - Albino R ata O rgan Weight and Ratio Data Three-Month Sacrifice Sum m ary of Mean Value Organ: Kidney o O rgan Weight (g) Male F em ale 4.03 2.49 4.37 2.62 4.00 2.61 4.00 2.52 Organ/Dody Weight Ratio (g /1 0 0 g) Male F em ale 0.782 0.802 0.838 0.911 0.747 0.782 0.779 0.793 O rgan/B raln Weight Ratio (g/g) Male F em ale 1.95 1.31 2.14 1.36 1.96? . 1.37 1.96 1.35 OI n i* i uw o LKKjOJ -X oo l I *6Z 0 0 Dietary Level of A ro clo r 1260 (ppm) C o n tro l 1 10 100 rOe o o Ul o i T A B LE XV TE ST MATERIA Li A ro c lo r 1260 Tw o-Y ear C hronic O ral Toxicity Study - Albino Rata Organ Weight and Ratio Data Three-Month Sacrifice Sum m ary of Mean Valuea Organ; Spleen O O rgan Weight (g) M ales Fem ales 0.766 0.578 0.846 0.SI6 0.838 0.632 0.718 0.556 O rgan/Body Weight Ratio (g /1 0 0 g) M ales Fem ales 0.150 0.184. 0. 161 0. 180 0.156 0.189 0.140 0.177 O rg an /B raln Weight R atio (g/g) M ales Fem ales 0.371 0.302 0.412 0.269 0.411 0.331 0.352 0.297 o i^ n o Vi t K * 'TT'VC- O Ci table xvi TES, T MATERIAL Aroctor 12i60 Tw o-Y ear Chronic O ral Toxicity Study - Albino Rata O rgan Weight and Ratio Data % Three-Monlh Sacrifice Sum m ary of Mean Values Organ: Gonada Dietary Level of A roclor 1260 (ppm | Control 1 10 100 O rgan Weight Organ/Body Weight Ratio (g) (g /1 0 0 g) M a le s _______ F e m a l e s __________ M a lc a Fem ales 3.60 0.114 0.698 0.0370 3.67 0.128 0.704 0.0450 3.03 0.121 0.715 0.0368 3.71 0.138 0.725 0.0429 O rgan/B rain Weight Ratio (g/g) M a le s ______ F e m a l e a 1.74 0.0599 1.60 0.0666 1.89 0.0636 1.81 0.0728 O ADM 0 0 3 7 5 r 0 % u TA B LE XVII $ T E ST M ATERIAL: A ro c lo r 1260 Tw o-Y ear Chronic O ral Toxicity Study - Albino R ats Organ Weight and Ratio Data Three-Month Sacrifice tN I o C r Sum m ary of Mean Values Dietary Level of A ro c lo r 1260 (ppm) Organ: Heart Organ Weight (g) M ales F emales Organ/Body Weight Ratio (g /1 0 0 g) M ales Fem ales O rgan/B raln Weight Ratio (g/g) M ales Fem ales l Cont rol 1.59 1.05 0.307 0.339 0.767 0.551 1 1.57 1.01 0. 300 0.353 0.765 0.526 10 1.58 1.04 0.296 0.311 0.781 0.546 100 o 1.52 0.946 0.298 0.297 0.743 0.505 ACM OC375 tI o OOO TA BLE XVII! TEST M ATERIAL: A ro clo r 1260 T w o-Y ear C hronic O ral Toxicity Study Albino R ats Organ Weight and Ratio Data a Three-Month Sacrifice Sum m ary of Mean Values Organ; Brain Dietary Level of A ro c lo r 1260 (ppm) O rgan Weight (g) M ales Fem ales O rgan/Body Weight Ratio (g /1 0 0 g) M ales F em ales Final Body Weights of Sacrificed Rata (g) M ales Fem ales Control 2.07 1.91 0.402 0.616 518 314 1 v, 2 . 0 5 1.92 0.192 0.672 523 287 10 2.04 1.91 0.380 0.572 536 337 too 2.05 0.400 0.591 517 321 O ii OJ f in t*\ 0CO* ACh 0C3755 O' Bf '-TEST J7 c. H istooatholocic Fir,dine a Histopathologic examination of tissues and organs ta k e n f r o m a l l a n im a ls f r o m b o th the c o n tro l an d 100 p p m g ro u p s was conducted. Tables XIX and XX list a ll histopathologic changes noted. All of the lesions noted in the m icroscopic exam ina tion of tissues w ere those of spontaneous d isease and are not unusual fo r the albino ra t. The m ost frequent findings w ere lesions in the trachea and lungs, indicating chronic murine pneumonia. These oc c u rre d in the control as well as the ra ts fed A ro clo r 1260. I O ADM C037 : 00 298 I Ie3l 33 TAB LZ XDC O T E S T M A TER IA L: A r o c lo r 1260 T w o-Y ear Chronic O ral Toxicity Study - Albino Rata Histopathologic Changes Three-Month Sacrifice Croup: Control N u4m b e r of Animals Organ Examined Findings 5 Males Trachea Lung U rinary bladder Tracheitis Chronic resp irato ry disease Hyperplasia 5 Fem ales Lung Chronie re sp ira to ry disease Average Incidence Grade 1 1.0 1 3 * 1 1.0 1.0 1.0 All other tissues and organs were normal histologically. Grading System 0 .5 * m inim al 1.0 * slight 2 .0 s mild 3 .0 * m oderate 4.0 * severe 5.0 * extrem e A0M 0 0 3 7 5 7 299 oc \ I iX (. B I rti r jfuu T A B L E XX T E ST M ATERIAL; A ro c lo r 1260 T w o-Y ear Chronic O ral Toxicity Study - Albino Rats Histopathologic Changes Three-Month Sacrifice G ro u p : 100 p p m N um ber of Animals Organ Examined Findings 5 Males Trachea Lung Tracheitis Chronic respiratory disease 5 Fem ales Traehea Lung Kidney Tracheitis Chronic resp irato ry disease Foeal lymphoid infiltration 'A verage Incidence G rid e 4 1.0 2 1.0 3 2 - 1 1.0 1.0 1.0 All other tissues and organs were norm al histologically. Grading 5vstem 0 .5 = m inim al 1.0 s slight 2 . 0 3 m ild 30 3 m oderate 4.0 severe 5.0 * extrem e O AGH 003756 00 300 i _7nr/ Hi**->i O i t ' i e a I snc. 40 2, Six-Month Sacrifice a. C ross P ath o lo g ieFindings No outstanding differences w ere noted between test and control anim als. b. Organ Weight and Organ to Bodv and Organ to Brain Weight Ratio Data The re s u lts of the statistical an aly ses conducted on absolute organ w eights, organ to body weight and organ to brain weight ra tio s a r e su m m a riz ed in Tables XXI through XXVI. O rgan w eights, organ to body w eight and organ to b ra in weight ratios disclosed several randomly occurring intergroup differences. The lack of any consistent dose related response and the absence of any deleterious histopathologic change confirm that these differences w ere n ot re la te d to the ingestion of A ro c lo r 1260. AC* 0C375 oc 301 oft* * oCo fo oo oO' o Dietary Level of A ro clo r 1260 (ppm) Control 1 10 100 o TABLE XXI T E ST M ATERIAL: A ro c lo r 1260 Tw o-Y ear .Chronic O ral Toxicity Study - Albino Rata Organ Weight and Ratio Data Six-Month Sacrifice Sum m ary of Mean Value Organ: Liver o O rgan Weight (g) M ales Fem ales 22.9 n .i 1 4 .8 15.1 19.0* |0 .5 * * 24.4 11.6 Organ/Body Weight Ratio (g /1 0 0 g) M ales Fem ales 3.74 3.77 2.92** 3..74 3.31* 3.08* 4.69* 4.03 O rgan/B raln W eight Ratio (g/g) M ales Fem ales 11.7 6.71 7 .0 5 * 7.66 9.45 6.07 12.3 6 . 17 '> S ta tis tic a lly s ig n ific a n t d if f e r e n c e a t the 95 p e r c e n t co n fid e n ce le v e l. Statistically significant difference at the 99 p e rc en t confidence level. Th i C ii is t Oo TABLE XXII TEST MATERIAL: A ro clo r 1260 T w o-Y ear Chronic O ral Toxicity Study - Albino R ate , O rgan Weight and Ratio Data * Six-Month Sacrifice Sum m ary of Mean Values Organ: Kidneys # Dietary Level of A roclor 1260 fppm) O rgan Weight (g) M ale______ F e m a l e s Organ/Body Weight Ratio (g /1 0 0 g) M ale______ F e m a l e O rgan/B raln Weight Ratio (g/fi) M a le a ______ F e m a l e s C o n tro l 4.04 2.79 0.664 0.806 2 r 06 1.43 1 3.64 2.73 0.716 0.674e. 1.73 1.42 10 oo 100 3.85 3.92 2.35 2.35 0.670 0.753 0.684* 0.814 uo> w * S tatistically significant difference at the 95 p e rc en t confidence level. 1.92 1.97 1.36 1.25 , 12 r nTr ,r u: m in AC 0C376 N AOM 0 0 3 7 t ooo Dietary Level of A ro c lo r 1260 (ppm) C o n tro l 1 10 100 T Ail LB XXIII T E ST M ATERIA L: A ro c lo r 1260 T w o-Y ear C hronic O ral Toxicity Study - Albino R ata O rgan Weight and Ratio Data Six-Month Sacrifice Sum m ary of Mean Valuea Organ: Spleen O rgan Weight (B> M alea Fem ales 0.752 0.684 0.754 0.908 0.836 0.566* 0.636 0.44J* Organ/Body Weight Ratio (g /1 0 0 g) M ales Fem ales 0 . 122 0.196 0. Hfl* 0.220 0. 146* 0.167 0.122 0.151 O rgan/B rain Weight Ratio <g/g) M ales Fem ales 0.384 0. 350 0.359 0.472 0.416 0.327 0.320 0.236* S ta tistically significant differen ce at the 95 p e rc e n t confidence level. * 4 ooo TA B LE XXIV TEST M ATERIAL: A ro clo r 1260 T w o-Y ear C hronic O ral Toxicity Study - Albino R ats O rgan Weight and Ratio Data * Six-Month Sacrifice Sum m ary of Mean Values Organ: Gonads Dietary Level of A ro c lo r 1260 Ippml O rgan Weight (g) M ales Fem ales O rgan/Body Weight Ratio (g/100 g) M ales F em ales O rgan/B raln Weight Ratio (g/gl M ales Fem ales C o n tro l 3.55 0.0960 0.593 0.0279 1.81 0.0492 1 3.23 0. 120 a 0.636 0.0297 1.54 0.0625* 10 3.56 0.0968 0.620 0.0287 , 1.77 0.0560 100 3.53 0.0710* 0.686 0.0250 o a S ta tistic a lly significant d iffe re n c e a t the 95 p e rc e n t confidence level* 1.77 0.0378* 6 ACM CC 37 QLZOO O <--) o trv * Dietary Level of -A roclor 1260 (ppm) Control 1 10 ICO TA B LE XXV TEST M ATERIAL: A ro clo r 1260 T w o-Y ear Chronic O ral Toxicity Study - Albino Rata O rgan Weight and Ratio Data Six-Month Sacrifice Sum m ary of Mean Valuea Organ: Heart O rgan Weight (g) M alee Fem alee 1.70 1.05 .45 1.12 1.63 1.05 1.49 1.06 O rgan/Body Weight Ratio (g /1 0 0 g) M ale. F enul.s 0.282 0.302 0.286 0.277 0.283 0.307 0.286 0.368** O rg an /B raln Weight Ratio (g/gl M alee Fcm alei 0.867 0.538 0.690 0.583 0.810 0.606 0.749 0.564 * S ta tis tic a lly aig n iflcan t d iffe re n c e a t the 99 p e r c e n t confidence le v e l. 0 i V l c a -C" ** * lr oO Dietary Level of A ro c lo r 1260 (ppm) C o n tro l 1 10 100 TABLE XXVI TEST MATERIAL A ro c lo r 1260 `T w o - Y e a r C h r o n ic O r a l T o x ic ity Study - A lb in o R a t a Organ Weight and Ratio Data Six-Month Sacrifice Sum m ary of Mean Values Organs Brain O rgan Weight (g) M ale a ______ F e m a l c e 1.96 1.95 2.10 1.92 2.01 1.73 1.99 1.88 O rgan/Body Weight Ratio (g/100 g) M a le a ______ F e m a l e s 0.320 0.560 0.412* 0.474* 0.349 0.382 0.504 0.650* Final Body W eights of Sacrificed Rata * (g) M a le a ______ F e m a l e s 613 348 509 405 575 . 343 522 289 * Statistically significant difference at the 95 p ercen t confidence level* o O oV_*14 O' n l OC 37 j t t d i t LoJ, 3 I T E S T J am -"Vi C. H i s t o p a t h o l o g i c Findi ngs Histopathologic examination of tissues and organs. ta k e n f r o m a l l a n i m a l s f r o m both the c o n tro l and 100 p p m g ro u p s w as conducted* T ables XXVII and XXVIII lis t all histopathologic changes noted. All of the lesions noted in the m icro sco p ic exam ination of tissues w ere those of spontaneous disease and are not unusual for the albino ra t. The m o st frequent findings w ere lesions in the trachea and lungs, indicating chronic m urine pneumonia. These occurred in the contro 4 a s w ell a s the r a ts fed A ro e lo r 1260. ( i O t ACM OC3706 0 ( 308 Blr T55 T ;Uc. 43 TABLE XXVII T E S T MATERIAL: A ro c lo r 1260 Tw o-Y ear C hronic O ral Toxicity Study - Albino Rats Histopathologic Changes *C Six-M onth S a c r if ic e Croup: Control N u m b e r o Animals Organ Examined Findings Average Incidence ' Grade 5 Males Trachea Lung Liver Skeletal muscle Tracheitis Focal tracheitis Focal pneumonitis H yperem ia Focal pericholangitis Focal inflammation 2 2 1 2 l l 1.5 1.0 0.5 1.0 1.0 1.0 5 Females Trachea Lung Liver Kidney Focal tracheitis H yperem ia H yperem ia Pericholangitis Focal inflammation i 1.0 2 2.5 1 1.0 1 0.5 l 1.0 All other tissues and organs were normal histologically. Grading System 0.5 a minimal 1.0 i slight 2 .0 a mild 3.0 * m oderate 4.0 * severe 5.0 extrem e 7 O ADM 0C37fc 309 00 I_J l TA B LE XXVIII T E S T M ATERIAL: A ro c lo r 1260 T w o -T ear C hronic O ral Toxicity Study - Albino Rats Histopathologic Changes Six-Month Sacrifice G ro u p : 100 ppm Number of Animals Organ Examined Findings Average Incidence G rade 5 Males Trachea Lung Liver Tracheitis Focal tracheitis Focal pneumonitis H yperem ia H yperem ia Focal pericholangitis. 2 1 4 4 l 1. 2 .0 0.5 1.0 1.5 1.0 0 .5 5 Fem ales Trachea Lung L iv er Kidney Focal tracheitis Focal pneumonitis H yperem ia H yperem ia Focal pericholangitis Focal nephritis 2 2 1 2 1 3 0.5 1.0 3.0 1.5 0.5 0.5 AH other tissues and organs w ere normal histologically. Grading System 0.5 m inim al 1.0 slight 2.0 a mild 3.0 a moderate 4.0 a severe 5.0 extrem e AOP C C37i jrtdtbU toel & I T E S T Jz&yusJ&uA, SO 3 T w e lv e -M o n th S a c r if ic e A* G r o s s P a th o lo c ic F in d in g s G r o s s p ath o lo g ic find Lag a am o n g t e s t a n i m a l s w e r e : significantly different fro m those noted in control anim als. b. Organ Weight and Ratio Data The m e an organ w eight and ra tio data collected a t :h tw elve m onth sacrifice a re p resen ted in Tables XXIX through XXXIV. * Statistically significant differences are designated by a ste risk s following the test group value. O rg an weights, organ to body w eight and organ to br; weight ratios disclosed several randomly occurring intergroup differences, The lack of any consistent dose related response and. the absence of any deleterious histopathologic change confirm that these differences w ere not re la te d to the ingestion of A roclor 1260. ACM 0 0 3 7 6 9 311 0 > o Dietary Level (ppm) Control I 10 100 oo T A tiL E XXIX TEST M ATERIAL: A ro clo r 1260 T w o-Y ear C hronic O ral Toxicity Study - Albino Rata Twelve-Month Sacrifice O rgan Weight and Ratio Data Sum m ary of Mean Values Organ: Liver Organ Weight <6> M ales Fem ales 22.1 15.5 17. ! 10.7* 20.0 13.6 28.8* 14. 1 Organ/D ody Weight Ratio (fi/100 g) M ales Fem ales 3.79 3.52 2.79** 3.01* 3.30 3.54 4.26 3.79 O rgan/B rain Weight Ratio <g/g) M ales Fem ales 10.8 7.72 9.23 5.76 10.8 7.28 13.4* 8.52 * Statistically significant difference at the 95 percent confidence level, Statistically significant difference at the 99 p erce n t confidence level, to % to ool*J ytimt u Dietary Level (ppm) Control 1 10 100 o TA B LE XXX TEST MATERIAL; A roclor 1260 Tw o-Y ear Chronic O ral Toxicity Study - Albino Rats Twelve-Month Sacrifice O rgan Weight and Ratio Data Sum m ary of Mean Values Organ: Kidneys O rgan Weight (g) M ales Fem ales Organ/D ody Weight Ratio (g/100 g) M ales F em ales O rgan/B rain Weight Ratio (g/g) M ales Fem ales 4.04 3.04 4.00 5.11* 3.19 2.58** 2.78* 2.81 0.692 0.625 0.660 0.758 0.738 0.724 0.736 0.755 1.97 2.06 2.16 2.38* 1.58 1.39 1.48 1.69 Statistically significant difference at the 95 percent confidence level. S tatistically significant difference at the 99 p e rc e n t confidence level. / 1t ** oo Dietary Level (ppm) Control l 10 100 TABLE XXXI TEST M ATERIAL: A ro clo r 1260 T w o-Y ear Chronic O ral Toxicity Study - Albino Rata Twelve-Month Sacrifice Organ Weight and Ratio Data Sum m ary of Mean Values Organ: Spleen O rgan Weight (g) M ales Fem ales O rgan/B ody Weight Ratio (g/100 g) M ales Fem ales 0.778 0.796 0.133 0.183 0.756 0.836 0.460+ 0.578 0 . 122 0.119 0.129* 0.150 0.785 0.474* 0.116 0.127* O rgan/B raln Weight Ratio (g/g) M ales Fem ales 0.380 0.408 0. 396 0.247* 0.457 0.365 0.308 0.284 * Statistically significant difference at the 95 p ercent confidence level. - vo ooIV G { 314 O Dietary Level fppml Cont rol 1 10 100 Oo T A B L E XXXII TEST MATERIAL; A roclor 1260 T w o-Y ear Chronic O ral Toxicity Study - Albino Rats Twelve-Month Sacrifice O rgan Weight and Ratio Data Sum m ary of Mean Value* Organ; Gonads O rgan Weight (g) M ales Fem ales O rgan/B ody Weight Ratio (g /1 0 0 g) M ales Fem ales O rgan/B rain Weight Ratio (g/g) M ales Fem ales 3.46 3.38 3.51 4.03** 0.116 0.103 0.112 0.088* 0.596 0.551 0.578 0.598 0.0275 0.0287 0.0294 0.0238 1.70 1.82 1.69 1.88 0.0574 0.0552 0.0584 0.0527 * Statistically significant difference at the 95 p e rc e n t confidence level. * Statistically significant difference at the 99 p ercen t confidence level. v => % o o Kj* 315 r O Dietary Level (ppm) Control 1 10 too Oo TABLE XXXIII T E S T M A T E R IA L : A r o c l o r `1260 T w o-Y ear Chronic O ral Toxicity Study - Albino R ats Twelve-Mouth Sacrifice O rgan Weight and Ratio Data Sum m ary of Mean Values Organ; Heart O rgan Weight (g) M ales Fem ales O rgan/B ody Weight Ratio (g/100 g) M ales Fem ales O rgan/B raln Weight Ratio (g/gl M ales F em ales 1.71 1.68 1.23 1.07 0.293 0.273 0.2B7 0.301 0.835 0.902 0.611 0.575 1.59 1.86 1.16 1.27 0.261 0.275 0.307 0.340 0.845 0.868 0.620 0.760 Iu-* Vo * CTi o c> nf S 1o T HS T 317 OOo Dietary Level (ppm ) Cont rol 1 10 100 T A B L E XXXIV TE ST M ATERIAL: A roclor 1260 Tw o-Y ear Chronic O ral Toxicity Study - Albino Rata Organ Weight and Ratio Data Twelve-Month Sacrifice Sum m ary of Mean V alues Org an Weight te) Males Fem ales 2.04 2.02 1.87 1.86 1.68 1.90 2.15 1.67** Organ: Brain Organ/Body Weight Ratio (g/IOOg) M ales F em ales 0.349 0.460 0.304 0.522 0.310 0.493 0.318 0.449 Final Body W eight of Sacrificed Rats (gram s) M ales Fem ales 584 439 616 356 607 385 675 372 a Statistically significant difference at the 99 p ercent confidence level. o 7 Oo f uO*' c. Hiatooathologic Finding a Histopathologic examination of tissues taken from each r a t s a c r i f i c e d f r o m the c o n tro l and 100 ppm g r o u p s a f t e r twelve m o n th s of testing was conducted. T ables XXXV and XXXVI lis t a il hisopathologic changes noted. All of the lesions noted in the m icroscopic exam ination of tis s u e s w e re those of spontaneous d is e a s e and a r e not unusual fo r the albino ra t. The m ost frequent findin gs w ere lesions in the trachea and lungs indicating chronic m urine pneumonia. These occurred in the contrc a s well as the ra ts fed A ro clo r 1260. AOK GG3776 318 ooc ?O TABLE XXXV TE ST M ATERIAL: A r o c lo r 1260 Two*Year Chronic Oral Toxicity Study - Albino Rata Histopathologic Changes Twelve-Month Sacrifice Group: Control N um ber of Animals 5 Males 5 Females Organ Examined TracheaLung U rinary Bladder Trachea Lung Colon Findings Incidence Tracheitis Chronic respiratory disease Calculus 1 2 1 Tracheitis Chronic resp irato ry disease P a ra site s 1 .* 1 1 Ave i Gra 1. 1. 1. 1. 1. 1. All other tissues and organs w ere norm al histologically. Grading Svstem 0 . 5 = minimal 1.0 = slight 2 .0 s mild .3.0 a moderate 4.0 severe 5.0 a extreme A0H 0 0 3 7 7 7 no 3 l9 8 1 T5 T ci**-**. ;U. 3* TABLE XXXVI TE ST M ATERIAL: A ro c lo r 1260 T w o-Y ear C hronic O ral Toxicity Study - Albino Rata Histopathologic Changes Twelve-Month Sacrifice C ro u p : 100 ppm N u m b e r of.. Animals 5 Males Organ Examined Traehea Lung Kidney Liver 5 Females Trachea Lung Findings Incidence Tracheitis Chronic resp irato ry disease Focal lymphoid infiltration Vacuolar change of random cells 1 3 1 l Tracheitis Chronic resp irato ry disease 1 1 Ave ra Grad 1.0 1.0 1.0 1.0 1.0 1.0 All other tissues and organs were normal histologically. Grading Svstem 0.5 = minimal 1.0 a slight 2.0 mild 3.0 * moderate 4.0 > severe 5.0 a extreme ADM O3770 nr 320 j B t >T S T jViC bQ 4. Final Sacrifice 4 C r o n P a th o lo g ic Findi ngs G ross pathologic findings among test animals were not significantly different from those noted in control anim als, b. Organ Weight and Ratio Data The m ean organ weight and ra tio data collected at the fin a l s a c r if ic e a r e p r e s e n te d in T ables XXXVII th ro u g h XI.1I. S ta tis tic a lly significant differences a re designated by a ste risk s following the test group value. A bsolute liv er weights and liv e r to body weight or liv e r to b r a in w eight r a tio s w e re sig n ific a n tly e lev a te d in those r a ts fed 100 p p m A ro c lo r 1260. O ther org an w eights, o rg an to body w eight and organ to b ra weight ratios disclosed several randomly occurring intergroup differences. The lack of any consistent dose related response and the absence of any deleterious histopathologic change confirm that these differences were n o t re la te d to the ingestion of A ro c lo r 1260 ACM 0 C 3 7 7 321 0( 1 S 3 1 * O i S V ^ '^ T 'v TA BLE XXXVII * T E S T M ATERIA L] A ro c lo r 1260 T w o-Y ear C hronic O ral Toxicity Study - Albino R ata Organ Weight and Ratio Data i Sum m ary ol Mean Valuea Final Sacrifice Organ: Liver Organ Woight Organ/Body Weight R atio O rgan/B rain W eight Ratio ictary Level (g) (g/IOQ g) (g/g) (ppm)____________M a le s _____________F e m a l e s ___________ M a le s ____________ F e m a l e s ____________ M a le s Fem ales onlrol 22.0 16. 3 3.47 3.05 10. 10 8. 42 1 22.0 16.6 3.40 3. 35 9.93 8.62 10 24. 1 18. 1 2. 87 + 3. 19 11.20 9. 26 00 29.9** 22.? ` 4 .67** 4.42M 13.40* 11.60* ' S tallstically significant difference at the 95 p e rc e n t confidence level. ' Statistically significant difference a t the 99 p e rc e n t confidence level. *j k> j o J nr C03 * )ictary Level (ppm) lonlrot 1 10 too T A B L E XXXLX TEST M ATERIAL: A ro clo r 1260 T w o-Y ear Chronic Ora! Toxicity Study - Albino Rata Organ Weight and Ratio Data Sum m ary of Mean Values Final Sacrifice Organ Weight M ales (6) Fem ales 1. 15 0.96 1. 17 0.72 1.09 0.83 0.89 0.75 Organ; Spleen Organ/Dody Weight Ratio (g / 100 g) M ales Fem ales 0 . 182 0. 181 0. 178 0. 148 0. H I 0.142 * 0 . 138 0. 146 i % t ** ' y | f O rgan/B rain W eight Ratio A (e/e) M ales Fem ales 0.520 0.492 0.529 0. 370 0.509 0. 428 0.400 0. 379 o o Lou 323 OO Dietary Level - fpnnil________ Control 1 10 100 TABLE XL TEST M ATERIAL; A roclor 1260 Tw o-Y ear Chronic O ral Toxicity Study - Albino Rata O rgan Weight and Ratio Data i Sum m ary of Mean Values Final Sacrifice Org an Weight M ales (g) Fem ales 2.52 0. 123 3. 56* 0. 140 Organ: Gonads Organ/Dody Weight Ratio (c/100 g) M ales Fem ales 0. 397 0.0222 0.550* 0.0317 3.36 0.177 0.400 0.033E 2. 97 0. 113 0.464 0.0229 ^ *> S ta tis tic a lly sig n ific a n t d if f e r e n c e a t th e 95 p e r c e n t co n fid e n ce le v e l. Cj ^ ADH 0 0 3 7 8 2 O O rgan/B rain W eight Ratio (g/fi) M ales Fem ales 1. 15 0.0628 1.61* 0.4718 1.56* 0.0900 1.33 0.0584 JU oo Dietary Level fppm) Control I 10 100 TABLE XLI T E ST M ATERIA L: A ro c lo r 1260 Tw o-Y ear Chronic O ral Toxicity Study - Albino Rata O rgan Weight and Ratio Data i Sum m ary of Mean Value Final Sacrifice Organ: Heart Organ Weight M ales () Fem ales I.B7 1.54 O rgan/Ilody Weight Ratio Cg/ 100 g) M ales Fem ales 0.294 0. 288 1.98 1.45 0. 304 0.294 1.90 1.58 0.2254* 0.279 2.00 1.49 0.313 0.288 >4 S ta tis tic a lly significant d iffe re n c e a t the 99 p e r c e n t confidence lev el. u> tuot C>*3 o o o O rgan/B rain W eight Ratio (e/g) M ales Fem ales 0. 860 0. 795 0.893 0.747 0.884 0.809 0.904 0.764 O vJ a* o i v c. Histopathologic Findings Histopathologic exam ination of tissues and organs ta k e n f r o m a l l a n im a ls s a c r i f i c e d a f t e r 24 m o n th s on te s t w as c o n d u cte d . Tables' XLIII through XLVI list all histopathologic chang n o te d / Table XLVII contains the re su lts of the histopathologic examinatio; of the u rin a ry bladders. S ig n ific an t liv e r in ju r y w a s found in a n im a ls fed 100 ppm A ro e lo r 1260 (T-IH). The compound a s s o c ia te d lesions consisted of vacuo change, focal H ypertrophy and focal h y p e rp la s ia . In addition# th e re w ere lesions of inflam m ation, necrosis, fibrosis and m inor degeneration in this group which, although seen in the controls and low er te st groups, w ere mo sev ere and frequent in the T-IH group. The vacuolar change was not seen in the control anim als b u t w as o b s e rv e d o ccasio n ally in liv e rs fro m T - I (lppm ) and T-CI (10 ppm) a n im a ls. It was frequent in the T-IH a n im als. This lesion is m orphologic indicative of fatty degeneration. F o rm a lin -fro z e n sections of liv e rs from re p resen ativ e anim als which displayed vacuolar changes were stained with Oil Red 0 to re v e a l the presen ce of fat. The v a cu o la r lesion in the cytoplas of these cells was positively identified as fat. The hypertrophie change found in the liver was foeal and often lim ited to the central lobular a re a w here groups of cells w ere swollen to two or three times their norm al size with c lea r pink homogenous cytopla The h y p e rp la sia was associated with the sam e cells and appeared to be an ACM C 0 3 7 6 * O' 326 U > < I h J I wo extension of the hypertrophic lesion. The h y p e rp la stic cell a were also usually hypertrophic. The m ost severe exam ples of hyperplasia appear- a s nodular grow ths with lim ited com pression of the surrounding n o rm al hepatic tissue. ' The m in o r le s io n s of d e g e n e r a t i o n , h e p a titis , d u c tu le ce c p ro life ra tio n , n e c ro sis and focal lymphoid in filtra tio n seen in the control" an im als and the T -Ia n d T-H groups are lesio n s of spontaneous d isea se an a r e not re la te d to the A ro d o r 1260. This lev el of liv e r disease is n o tu n u in old anim als. H yperplasia of the u r in a r y b la d d e r was found in an anim ; f r o m the c o n tr o l g ro u p . T his finding w a s n o t p r e s e n t in any of the r a t s fee o A ro c lo r 1260. All of the other lesions in o th er tissu e s found in these anirr a r e re la ted to spontaneous disease and they a r e not unusual for old r a ts . Donovan C o rd o n , C. V. M . , F h . D. D iplom ate, A m erican College of V eterinary Pathologists O ACM C037 OC 3 2 7 n ooo Dietary Level (ppm) Control 1 10 too TABLE XL!! . T E S T M ATERIA L: A ro c lo r 1260 T w o-Y ear Chronic O ral Toxicity Study - Albino R ata * O rgan Weight and Ratio Data i Sum m ary of Mean Values Organ Weight (g) M ales Fem ales Final Sacrifice Organ: Brain Organ/Body Weight Ratio (g/100 g) M ales Fem ale Final Body Weight of Sacrificed Animals (gram s) M ales Fem ales 2.18 1.95 0. 342 0. 362 642 551 2.21 2. 15 1.94 l.% 0. 343 0.255** 0.407 0.347 654 845* 525 584 2.22 1.95 0.346 0.378 641 644 I I I I 1 * f Statistically significant difference at the 99 percen t confidence level. ADM 0 0 3 7 8 6 OO'' 328 6 Z t 00 OO TAJJ LG X LIII TEST M ATERIAL: A roclor 1260 Tw o-Y ear C hronic O ral Toxicity Study - Albino Rate Histopathologic Changes Final Sacrifice Num ber of A nim als II M ales * D X o o u> Ob -I Organ Examined Heart Trachea Lung Liver Stom ach Spleen Kidney Testes Prostate Adrenal Gland Drain Eye Croup: Control Findings Chronic focal m yocarditis T rac h e itis Chronic respiratory disease Focal degeneration Ductile hyperplasia Chronic gastritis Hem osiderosis Chronic nephrilis Testicular degeneration P e ria rte ritis Inflam m ation Telangiectasis Nodular hyperplasia Focal fatty change Focal encephalitis Inflam m ation Incidence 3 3 9 1* 2 1 3 8 2 2 1 1 1 1 2 O A verage G rade 1.0 1.0 1.0 1.0 1.0 1.0* 1.0 1.0 1.0 1.0 1.0 1.0 1.0 1. 0 1.0 1.0 o4 OO i TABLE XLIII continued TEST M ATERIAL: A ro clo r 1260 T w o-Y ear C hronic O ral Toxicity Study - Albino R ats Histopathologic Changes Final Sacrifice Group: Control Number of Animals Organ Examined Findings Incidence 330 H Females O ot> n oLou CD Heart Trachea Lung Liver Stomach Colon Spleen Uterus Kidney Ova ry Adrenal Gland Salivary Gland Chronic focal m yocarditis Tracheitis Chronic respiratory disease Focal lymphoid infiltration Ductile hyperplasia Focal hepatitis Non-lipid vacuolar change Chronic gastritis Acanthosis - squamous epithelium Parasites H em osiderosis M etritis Chronic nephritis Focal lymphoid infiltration Cyst Telangiectasia Nodular hyperplasia Chronic fibrosis and atrophy All o lh c r tis su e s and organs were n o rm al histologically. 7 9 n 3 2 1 2 3 5 1 4 3 6 1 1 9 4 1 O A verage G rade 2.0 1.0 1.0 1.0 1.0 1.0 ' 0. 5 1.0 1.0 1.0 1.0 2. 0 1.0 1.0 1.0 1.0 1.0 1.0 331 OO TABLE XLIV T E S T M A TER IA L: A ro c lo r 1260 Tw o-Y ear Chronic O ral Toxicity Study - Albino Rale Histopathologic Changes Final Sacrifice Group; I ppm * Number of A n im a la ________ O r g a n E x a m in e d 11 M ale s Heart Trachea Lung Liver Pancreas Stomach Spleen Lymph Node Kidney Testes Prostate Scminai Vesicle Findings Chronic focal m yocarditis Tracheitis Chronic respiratory diseaso Focal hepatitis Focal scarring H em osiderosis Chronic focal pancreatitis Acute focal g astritis H em osiderosis Congestion Chronic nephritis Testicular degeneration P e ria rte ritis Inflammation Inflammation Incidence 4 1 10 1 1 I 2 1 1 1 7 1 2 4 1 o o O A verage G rade 1.0 1.0 2.0 1.0 1. 0 1.0 1.0 3.0 1.0 1.0 2.0 2. 0 1.0 2. 0 2. 0 r TABLE XLIV continued T E ST M ATERIA L: A ro c lo r 1260 T w o-Y ear Chronic O ral Toxicity Study - Albino R ate Histopathologic Changes Final Sacrifice Number of A n im a ls ________ O r g a n E x a m in e d Group: 1 ppm ' A verage F in d in g s _____________________________ In c id e n ce______________ G r a d e 11 Males Adrenal Gland Nodular hyperoplasia Focal fatly change 2 1.0 2 1.0 14 F e m a l e s oO uu>> to nX> IooaJ H eart Trachea Lung Liver Stomach Chronic focal m yocarditis Chronic focal m yocarditis Tracheitis Chronic respiratory disease Focal fibrosis Focal lymphoid infiltration Ductile hyperplasia Congestion Vacuolar change (fatty) Chronic gastritis Acanthosis 3 1 9 n i i i i i i 2 1.0 3.0* 1.0 1.0 1.0 1.0 0. b 1. p 2. 0 1.0 1.0 000 333 o Number of Animala l -1 F e m a l e s P X o oU->4o TABLE XLIV continued T E ST M ATERIA L: A ro c lo r 1260 Tw o-Y ear Chronic O ral Toxicity Study - Albino Rate Histopathologic Changes Final Sacrifice Group: I ppm * Organ Examined Stomach Colon Spleen Lymph Node Kidney Uterua Adrenal Gland Findings U lceration and acanthosis P a ra site s Hemosiderosis Hyperplasia Congestion Chronic nephritis Vacuolar change (fatty) M etritis Telangiectasis Nodular hyperplasia Congestion Incidence 1 1 11 I 1 1 1 4 9 3 i All other tissu es and organa w ere n o rm al histologically. 0.5 s minimal Grading System 0 = mild an Q A verage G rade 1. 0 1.0 1.0 1,0 1.0 2.0 . 1.0 1.0 1.0 1.0 1.0 oo Number of Animals 7 Males * TABLE XLV T E ST M A TERIA L: A ro c lo r 1260 T w o-Y ear Chronic O ral Toxicity Study - Albino Rata Histopathologic Changes Final Sacrifice, . G ro u p : 10 ppm * Organ Examined Heart Trachea Lung Liver Stomach Spleen Kidney Prostate Adrenal Gland Finding* Chronic focal m yocarditis Tracheitis Chronic respiratory disease Suppurative pneumonia Vacuolar change (fatly) Chronic gastritis Acanthosis H em osiderosis Chronic nephritis Inflammation Nodular hyperplasia Incidence 3 2 5 1 2 1 2 3 6 2 1 =5 Cj 4 i M Female* __ n o tu Heart Trachea Lung Chronic focal m yocarditis Thraclieitits Chronic respiratory disease 3 B 14 A verage G rade 1.0 1.0 1.0 3.0 1.0 1.0 1.0 1.0 1.0 1-0 . 1.0 1.0 1.0 1.0 4 r r `T^nr^ncrr Number of Animals N Fem ales o>X ot-4u4) 9 TABLE XLV continued T E ST M A TER IA L: A ro c lo r 1260 T w o-Y ear C hronic O ral Toxicity Study - Albino R ats Histopathologic Changes Final Sacrifice G ro u p : 10 p p m # Organ Examined Liver Pancreas - Stom ach Spleen Kidney Uterus Pituitary Adrenal Gland Findings V acuolar change (fatty) Focal hyperplasia Chronic focal pancreatitis P e ria rte ritis Acanthosis. H em osiderosis Chronic nephritis M etritis Congestion Telangiectasis Nodular hyperplasia Congestion Incidence 1 1 1 12 10 . 7 1 7 I 1 All other tissu e s and organa w ere n o rm a l histologically. r!-- m tn . ? A verage G rade 1.0 1.0 3.0 3.0 . . 1.0 t.o 2.0 LO 1.0 1.0 1.0 tn 335 n O Number of Animals 10 M ale s u> oa\j o> oo ui ---- i. * TA B LE XLV1 T E S T M ATERIA L: A ro c lo r 1260 T w o-Y ear Chronic O ral Toxicity Study - Albino Rato Histopathologic Changes Final Sacrifico G ro u p ; 100 p pm Organ Examined Heart Trachea Lung Liver * Pancreas Stomach Spleen Kidney 1 Findings Chronic focal m yocarditis Tracheitis Chronic respiratory disease Congestion Vacuolar change (fatty) Focal degeneration Congestion Centrolobular hypertrophy Telangiectasis Chronic focal pancreatitis Inflam m ation Acanthosis H em osiderosis Focal lymphoid infiltration Calcified concretions in pelvis Incidence 5 6 9 1 2 1 3 2 1 2 2 2 7 1 1 A verage G rad 1.0 1.0 2.0 1.0 2 .0 1.0 2.0 1.0 1.0 1.0 2.0 1.0 1.0 1.0 1.0 1.0 r- Number of A nim als 10 M ales TABLE XLV1 continued T E S T M A TERIA L: A ro c lo r 1260 T w o-Y ear Chronic O ral Toxicity Study - Albino Rate Histopathologic Changea Final Sacrifice C ro u p : 100 ppm* Organ Examined Kidney (continued) Testes Prostate Thymus Adrenal Gland Findings Calcified concretions in pelvis A sperm ia Inflammation Cyst Telangiectasis Nodular hyperplasia Incidence 1 1 1 1 3 1 15 F e m a le s oo Co Co VI " n x onio H eart Trachea Lung Liver Chronic focal m yocarditis Tracheitis Chronic respiratory disease Congestion Edema Vacuolar change (fatty) 5 8 11 1 1 5 O l% .1 I A verage G rade 1.0 1.0 1.0 1.0 1.0 1.0 Ii l \ J a I 1.0 i.O 1.0 1.0 1.0 1.0 --4j r Number of Animals 15 F e m a le s > r nob> *r> o TABLE XLVI continued T E ST M A TER IA L: A r o d o r 1260 Tw o-Y ear Chronic O ral Toxicity Study - Albino Rata Histopathologic Changes Final Sacrifice G ro u p : 100 p p m ' Organ Examined Liver (continued) Pancreas Spleen Kidney Uterus Pituitary Adrenal Gland Parathyroid Findings Incidence Focal hyperplasia Chronic inflammation of liver capsule Centrolobular hypertrophy N ecro sis Ductile cell hyperplasia, portal fibrosis and w idespread eosinophil inclusions in heputucytu cytoplasm Chronic focal pancreatitis and periarteritis Hem osiderosis Chronic nephritis M etritis Congestion Telangiectasis Nodular hyperplasia Congestion Hype rplasia 4 1* 5 2 1 1 9 8 7 1 6 1 1 1 All o lh r ti s s u e * and o r g a n s w e r e n o r m a l hintnlo****!' A verage G rade 1.0 1.0 1.0 2.0 1.0 3.0 2.0 1.0 1.0 1.0 1.0 1.0 1.0 1.0 338 r r ooo Group C T-l * (1 ppm) T-H (10 ppm) T-1II (100 ppm) T*ABLE X L V H TEST MATERIAL: A roclor 1260 Tw o-Y ear Chronic O ral Toxicity Study - Albino Rata Histopathologic Changes of the U rinary B ladder Final Sacrifice Number Sex Examined F in d in g s'* Incidence M 11 F 14 P ro tein ac eo u s plug (34) Focal epithelial h y p erp lasia (62) 1 1 M 10 F II M7 F 14 P ro te in a c e o u s plug (86. 105. and 107) None P ro tein aceo u s plug (187) None 3 - i 1 - M 10 F 14 P ro tein aceo u s plug (271) None 1 - A verage G rade 1.0 - m - * Individual anim al num bers a re shown in p arentheses. Grading System O 1 . 0 - minima! or trace 4.0 = severe 2 . 0 --m ild 5.0 = extreme 1.0 = moderate ACM CC37 O IW - I S J I * G. Detailed Description of Tumor Findings Tum or data for individual anim als including location, weight and pathologic classification a re presented in T ab les XLVIH through LI. None of these tum ors could be re la te d to the ingestion of A rocloi 1260 and a re co n sid ered n o rm al for a ran d o m population of ra ts this age. 3 AQM 0031 340 0( C O Animal ' u m b er m! Sex Location * 1-M Abdomen 2-M Pancreas i6-F 17- F B -F Abdomen i Lymph node I Abdomen 50-F Abdomen 52-F > 39-F . 5 64-F o O ^J Abdomen Abdomen Ad rnal gland Abdomen O t TABLE XLVIII TEST M ATERIAL; A roclor 1260 Tw o-Y ear Chronic O ral Toxicity Study - Albino Rata D escription of Tum or Findings Group; Control W eight (gram s) Stxe (cm) . C lassificatio n - M amm ary fibroadenoma 11.76 - A denocarcinom a 6 4 . 9B 35.09 8x1x2 5x2x2 M am m ary fibroadenoma , M am m ary fibroadenoma - - Reticulum cell sarcom a - 2 x 2 x 1 M am m ary fibroadenoma 88.92 2x2x3 ^s - 13 x 11 x 4 M am m ary fibroadenoma M am m ary fibroadenoma M amm ary fibroadenoma Phcochrom ocylom a F ibroadenom a O M onths on Test 24 24 24 24 24 7 24 24 24 341 lu O Animal Number and Sex 9')-M 127-F 128-F 138-F Ml -F 151-F Mc* 152-F * o T an Location Pituitary Pituitary Abdomen Pituitary Abdomen Abdomen Piluilary Ovary Abdomen Abdomen O TABLE XUX m TEST M ATERIAL: A ro clo r 1260 Tw o-Y ear Chronic O ral Toxicity Study - Albino Rata Description of Tum or Findings Group: 1 ppm Weight (gram a) 25.00 .6 0 .8 0 * '' 20.07 - M . 89 540.00 Size (cm) C laeaiflcatlo n - /^`C h ro m o p h o b e a d e n o m a 6x3x1 5x6x2* / Chromophobe adenoma 1 M ammary fibroadenoma t Chromophobe adenoma *" M a m m a r y f ib r o a d e n o m a M am m ary fibroadenoma - t Chromophobe adenoma - /^Adenoma - M ammary fibroadenoma 1 3 x 1 5 x 2 * M ammary fibroadenoma O M ontha on Test 20 24 21 ,i 24 24 24 22 oO Animal Number and Sex * 205-F 208-F 213-F 216-F 220-F 221 - F CJ U) 1 o X TABLE L TEST M ATERIAL: A ro clo r 1260 T w o-Y ear Chronic O ral Toxicity Study - Albino Rata D escription of Tum or Findings G roup: 10 ppm Location W eight ' (grams) Abdomen 190.00 Pituitary - Abdomen - Right shoulder 138. 00 Abdomen Pituitary 9.57 - S ubm axlllary region Abdomen 14.44 29.0 Size <c TM> 9x5x4 - *- - _ 2x3x1 3x5x2 , Classification i' ` M a m m a r y f ib r o a d e n o m a i- C h ro m o p h o b e a d e n o m a i M am m ary fibroadenoma , u Subcutaneous tum or ' `M a m m a r y f ib r o a d e n o m a ; Cliomophobe adenoma i Fibroadenoma . i M ammary fibroadenoma O M onths on Teat 24 24 24 23 24 24 ~ Animal Number and Sex V 2 2*- F 233-F 234-F Location Left fem oral region Pituitary Abdomen Pancreas TABLE L continued TEST M ATERIAL: A ro clo r 1260 T w o-Y ear Chronic O ral Toxicity Study - Albino Rate D escription of Tum or Findings G roup: 10 ppm W eight (urania) 56.00 Size (cm) 6x8x2 C lassificatio n ^S ubcutaneous tumor 6 0 . 82 6x4x3 K Chromophobe adenoma ^ M ammary fibroadenoma a*I s le t c e l l a d e n o m a M onths on Test 6 24 24 1 !1 * \ 344 O o o 1 Ui Cl) * Animal Number and Sex 2-H-M 264-M 2B1-F 291-F 299-F 301,F. Location Pancreas Pancreas Abdomen Liver Abdominal cavity s Liver t TABLE LI TEST MATERIAL) A ro clo r 1260 Tw o-Year*Chronic O ral Toxicity Study - Albino Rata D escription of Tum or Findings G roup: 100 ppm W eight (gram e) 35.04 50.75 i. Size (cm) * 6x4x2 6x4x3 - C laesificatio n 1 Adenoma f^lslet celi adenoma ^ M a m m a ry fibroadenoma Lymphos a re orna s / Fibroma Lymphos a re orna u u\ *o o C) Qu}> 3 t* M onths on Test 24 21 24 23 24 23 i nr n Animal Number and Sex '3 0 2 -F 30A-F 311-F 312-M 314-F % 1 Location Lymph node Pituitary Abdomen Pituitary Abdomen TA B LE LI continued TEST MATERIALS A ro clo r 1260 T w o-Y ear Chronic O ral Toxicity Study - Albino Rats Description oi Tum or Findings G roup: 100 ppm W eight (gram s) 14. 00 26.00 Sire _ 1cm) 4x3x2 1x2x1 5x4x3 C lassificatio n L ym phosarcom a (/# C h ro m o p h o b e a d e n o m a 'Mammary fibroadenoma . 1M am m ary fibroadenom a ''chrom ophobe adenoma M ammary fibroadenoma i j i M onths on Teat 24 24 24 24 24 OACi o o T a o io ^, ) 11/ d 8 K T E S T ;'..* R EPO R T TO MONSANTO COMPANY TW O-YEAR CHRONIC ORAL TOXICITY WITH AROCLOR 1254 IN ALBINO RATS NOVEMBER 12, 1971 IB T NO. B72?8 L Introduction At the re q u e st of M onsanto Company, a tw o -y e a r chronic o ra l toxicity study was conducted using albino ra ts to determ ine the potential toxicity of A roclo r 1254, Lot No. AK-38. The following re p o rt re p re se n t: the re su lts of this investigation. e O ADM 00354 00 347- 3 l C J T S S T tn riv u fi' Z H. Sum m ary A tw o-year chronic toxicity study was conducted using albino ra ts fed d ie ts c o n ta in in g 1 (T -I), 10 ( T -11} o r 100 (T-IIIJ ppm A r o c l o r 1254. Food consum ption w as not a lte red in any te st group. Body weight gains of t e s t an d c o n t r o l a n im a ls did n o t d iffe r a f t e r 3 o r 12 m o n th s of t e s t i n g . A f t e r 24 m o n th s f e m a le s fed 100 p p m A r o c l o r 1254 had gain ed le ss w e ig h t than the c o n tro l fe m a le s. Weight gains of the m a le ra ts did not d iffe r f r o m the c o n tro ls . The num ber of a n im a ls dying and the tim e at w hich d e a th o ccu rred did not differ among test and control groups. H em atological and clinical blood chem istry studies conducted after 3 6, 9. 12, 13, o r 24 m o n th s did n o t r e v e a l an y a f f e c t s r e l a t e d to A r o c l o r 1254. U rin e an aly ses failed to rev eal any d ifferen ce between te st and -control rats. 1 A t s a c r i f i c e afLer 3, 6, o r 12 m o n th s on t e s t , o r g a n w e ig h ts , o r g a n eo body w eight and org an to b rain w eight ra tio s d is c lo s e d s e v e ra l ra n d o m ly occurring intergroup differences*. The lack of any consistent dose related response and the absence of any deleterious histopathologic change confirm that these d ifferen ces w ere not related to the ingeseion of A roclor 1254. II A t the fin al s a c rific e a fte r 24 m onths on t e s t , the absolute liv e r w eig h t <1 and liver to body weight or brain weight ratios w ere significantly elevated in b oth T-IIX m a le s and fe m a le s . H istologic e x a m in a tio n s of the l i v e r s 1 f r o m the T -IH group rev ealed s e v e ra l animal.' v-ilh v a c u o la r change. AOM 0 0 3 5 4 0C 348 UIO *T 1 T j 3 T h is le s io n is m o rp h o lo g ic a lly indicative- of fa tty d e g e n e ra tio n . S p e c ific fat sta in s c o n firm e d the p re s e n c e of fat in th ese v a cu o les. F o c a l h y p ertro p c and focal h y p erp lasia w ere also found in the liv ers from anim als fed A r o c l o r 1254. H y p e rp la sia of the u rin a ry blad d er was found in anim als fro m the contrc g ro u p and fro m eaeh of the te s t g ro u p s. This h y p e rp la sia was u su a lly associated with cystitis. None of the tum ors found could be related to the ingestion of A ro c lo r 1254 and a r e c o n s id e re d n o r m a l for a ra n d o m po p u la tio n of r a t s th is a g e . Respectfully submitted, INDUSTRIAL BIO-TEST LABORATORIES, INC. R ep o rt p re p a re d by: Philip Sm ith, B. S. A ssistant Toxicologist Rat Toxicity R ep o rt approved by: Rat Toxicity Faul L. W right, Ph. D. Section Head, Toxicology Mi L . Kcphiir.ur, P U .^ . M anner, Toxicology N o v em b er 12, 1971 bb:hp AOM 0 C 3 OC 3 4 9 3 IO TEST no. fa& 4 III. P ro ced u re A. Experim ental Animals The anim als employed in the test w ere C harles River Strain* alb in o r a ts . P o u r hundred ra ts (200 m ale and 200 fem ale) w ere s e le c te d for the e x p erim en t, ear-punched with the anim al num ber assigned and housed individually in standard w ire-bottom ed steel ra t cages. E ach cage bore a color-coded card identifying the rat w ith respect to p ro ject num ber, dietary level assignm ent, individual num ber and sex. B. O rg a n iz a tio n of G roups A s t r u c t u r a l o u tlin e of the e x p e r i m e n t is g iv e n in T ab le 1. TABLE I TEST MATERIAL: A ro c lo r 1254 T w o-Y ear Chronic O ral Toxicity Study - Albino Rats O utline of E x p e rim e n t Grouo N um ber of A nim als Male Fem ales Dietary Level (oom) C o n tro l T-I T-H 50 50 None A dm inistered 9 50 50 1 50 50 10 T -m 50 50 100 * C harles R iver Breeding Laboratories, North Wilmington, M ass. an C0354 ( D 350 i-1'.uzL B I O T E S T ci, ;Vui 5 C D i e t s , F e e d i n g and Food C o n s u m p t i o n A ll d iets w ere prepared in the c e n tra l diet room of this laboratory. The basic ration from which all diets were constructed was a standard, pulverized stock ra t ratio n -. The diet for any given te st group was p re p a red by blending the calculated amount of A roclor 1254 w ith a pre-w eighed portion of the stock ratio n in a H obart m ixer. F re s h diets were prepared each week and every rat was offered an am ount of food sufficient for ad libitum feeding. Food consumption was recorded for five rats of each sex In ev ery group weekly for the first three months and monthly for the next nine months. T hereafter, periodic spot checks w ere made. D. B odv W eig ht s and V"eight G a in s Initially, the body weight of each ra t in every group was d e t e r mined and recorded. T hereafter, individual weighings w ere made w eekly f o r the f i r s t 13 w e e k s and m o n th ly t h e r e a f t e r u n til the c o n c lu s io n of th e investigation. E. M ortality and Reactions Checks for m ortality and abnorm al behavioral reactions w ere m ade Ii daily throughout the investigation. * j F. Hem atologic Studies and Urine A nalyses ~ 'i *| t Blood studies, including determ inations of hemoglobin co n cen tra tion, hem atocrit value, erythrocyte count and both total and differential * Purina Rat Chow, R alston Purina Co. St. Louis. Mo. AO* m -351 QQ3- I i l !0 7 a 3 T J*uz. 6 leukocyte counts, w ere conducted upon five m ales and five fem ales from b oth the c o n tr o l and 100 ppm g ro u p s. U rine analyses for the presence of glucose, albumin, m ic ro scopic e le m e n ts, and determ inations of pH and specific gravity w ere c o n ducted upon urine sam ples at the sam e tim e intervals and from the sam e anim als as those employed for the blood studies. T h e s e stu d ies w ere conducted a fte r 3, 6, 9, 12, 18, and 24 months of feeding. C. C linical Blood C hem istry Studies D eterm in atio n s of blood u re a n itro g en concentration (BUN), serum alkaline phosphatase activity (SAP), fasted blood glucose concen tration and seru m glutam ic-pyruvic tran sam in ase activity (SGPT) w ere conducted a fte r 3, 6, 9, 12, 18, and 24 m onths of feeding upon the sa m e ra ts as w ere the routine blood studies. H. Pathologic Studies When an anim al succumbed during the test period, a gross autopsy was perform ed. In those case when p ostm ortem autolysis was not a d vanced, representative tissues w ere taken and preserved in ten percent form alin solution far possible future histopathologic study. A f t e r 3 , 6 , and 12 m o n th s of t e s t i n g , fiv e a n i m a l s of e a c h s e x f r o m each group w ere sacrificed and subjected to com plete gross pathologic exam inations. A bsolute organ weights w ere re c o rd e d and organ to body weight and organ to brain weight ratios computed. The following organs w ere ADM 0 0 3 5 Ol 352 B I 0 T E S T Jz^yu^ytLtd, jUc included: liver, kidneys, spleen, gonads, heart and brain. These data w ere then subjected to statistic a l analysis. An A nalysis of V ariance was conducted first and significant effects disclosed by that treatm ent w ere f u r t h e r s t u d i e s b y Mt n - t e s t s . Com plete m icroscopic examinations w ere conducted upon the t i s s u e s and o r g a n s f r o m a ll c o n tro l and 100 p p m g ro u p a n im a ls s a c r i f i c e d a f t e r 3, 6, o r 12 m o n th s of te s t i n g . The fo llo w in g t i s s u e s and o rg a n s w ere included in the examinations: heart, tra c h ea , lungs, liver, p ancreas, esophagus, stom ach, sm all intestine (duodenum, jejunum and ileum), caecum colon, spleen, lymph nodes, kidneys, u rin ary bladder, gonads, prostate gland, sem inal vesicles, uterus, pituitary gland, adrenal glands, salivary glands, thyroid gland, parathyroid glands, skeletal m uscle, sternum , bone, peripheral nerve, spinal cord'and brain (cerebrum , cerebellum and pons). The aforem entioned tissues w ere p rep ared and stained with Hem atoxylin-Zosin stain. A fter two y ears of feeding, all surviving anim als w ere sacrificed and subjected to complete gross pathologic exam ination. The absolute weights of the liv er, kidneys, spleen, gonads, h e a rt and brain w ere recorded immediately after sacrifice. M icroscopic examinations were conducted upon tissues and organs from selected anim als dying during the ex p erim en t and upon all anim als fro m the final sacrifice. ACM OC 355 n r tci '*al O K T E S T J i^yu sJ a 'ic i, 5*to. 8 T issu es and organs examined w ere the sam e as those previously m entioned. L Tum or Incidence and C lassification A tabulation of the incidence of tu m o r fo rm atio n in each group w a s m ad e a t the conclusion of the in v e stig a tio n . In ad d itio n , tu m o r d a ta for individual anim als including location, w eight, size and pathologic classification were recorded. i I t I AOK G 03552 . 354 nr 3K I S0 * ,V ii 7 IV. R esults A. Body Weights and Weight Gain Tho m ale and fem ale body weight data a re a ssem b led in Table II. The d a t a in th is ta b le a lso inclu d e the 3. 12 an d 24 m o n th to ta l w e ig h t gains. 1 O ADH 0C 3553 355 m n O Dietary Level of A ro clo r 1254 (ppm) Sex Control M F 1 .M F 10 M F 100 M F >a 3: o0 l*J 01 Ul u o TA B LE II T E S T M A TER IA L; A ro c lo r 1254 T w o-Y ear C hronic O ral Toxicity Study - Albino Rats Body Weight and Total Weight Gain Data Sum m ary oi Mean Values 0 1 2 34 Body Weight (gram s) Week; 56 78 9 10 1 1 12 13 133 203 245 313 362 373 391 409 424 440 440 449 467 474 136 178 190 219 244 254 259 265 270 276 281 278 290 288 133 206 253 322 364 373 396 4 2 5 ' 4 36 457 470 483 490 509 136 171 190 213 235 246 248 250 259 270 270 274 279 285 133 209 251 325 375 394 402 410 432 453 470 488 494 508 136 171 194 220 243 263 2119 293 281 272 276 274 280 288 133 207 260 324 365 375 295 419 42U 440 450 469 480 486 136 174 197 223 242 254 276 290 281 268 270 274 277 280 90-D ay Total Weight Gain (gram s/rat) 3 11 152 376 119 376 152 * 353 114 Of) 3 5 6 O Dietary Level of A ro c lo r 1254 (pirn) Sex Control M F 1M F 10 M F 4 498 297 522 289 529 300 100 3~ M 495 F 292 u Ul -4 O It oo * * --*, O T A BL Et II co n tin u ed T E S T M A TER IA L: A ro c lo r 1254 Tw o-Y ear Chronic O ral Toxicity Study - Albino Rata Body Weight and Total Weight Gain Data Sum m ary of Mean Values 56 Body Weight (grama) Mtmlli: 7 8 9 10 II 533 565 593 622 642 667 688 310 322 333 315 359 374 383 550 581 604 633 658 680 715 295 307 322 333 340 354 , 367 567 593 . 621 642 667 681 700 308 321 330 341 354 367 380 521 541 562 586 608 627 642 300 306 316 321 326 332 340 12-M onth Total Weight Gain 12 ( g r a m s / r a t ) 701 568 390 254 725 592 3H0 244 726 593 392 256 664 531 347 211 QO dietary Level of A ro c lo r 1254 Sex 'Jon l ro t M F 1M F 10 M F 100 M F T A B L E II co n tin u ed T E S T M ATERIA L! A ro c lo r 1254 T w o-Y ear C hronic O ral Toxicity Study - Albino Rata Body W eight and Total W eight Gain Data Sum m ary of Mean Values 13 14 15 16 Body Weight (gram s) Month; 17 IH 19 20 21 22 23 24 - M onth Total Weight Gain 24 ( g r a m s / r a l ) 728 746 759 768 769 767 760 ' 758 739 726 6 h? 642 426 444 466 481 500 515 533 547 551 550 564 551 509 415 760 780 807 800 802 795 786 769 750 728 715 656 401 426 476 483 493 510 521 532 537 542 553 540 523 404 746 774 824 7 HO 804 ' 826 823 821 813 800 767 762 421 433 485 466 475 481 485 492 298 . 500 471 500 629 364 6GB 714 735 730 733 726 716 700 6B5 652 628 656 360 367 392 374 381 394 393 398 401 400 395 41 I 523 275 ^ ,|J S t a tis tic a lly s ig n ific a n t a t the 99 p e r c e n t c o n fid e n c e l i m i t . u> U1 00 AOH 003555 B. Food Consumption The data obtained fro m the food consum ption m e a su re m e n ts c o n d u c te d d u r i n g th e f i r s t 12 m o n th s of fe e d in g a r e s u m m a r i z e d in T a b l e III. T h e se data revealed no significant difference betw een te st and control ra ts . The periodic checks conducted during the rem ain d er of the investigation also revealed no outstanding d ifferen ces betw een te st and control anim als. ADrt GC355 OC 359 'I T A B LE III T E ST MATERIAL: A ro clo r 1254 Tw o-Y ear Chronic O ral Toxicity Study - Albino Rata Food Consumption Data S u m m ary of M ean Valuei Food Consumption Dietary Level of A ro clo r 1254 Inpn) Sex 1 2 3 4 (gram s/rat/seven days) Week: 56 789 Average Food C onsum ption 10 11 12 13 (graim s / r a t / s e v e n d Coni rol M 94 160 175 163 137 137 225 197 204 188 191 194 196 F 73 140 144 107 104 133 208 154 165 151 146 150 151- 174 140 1 M F 112 87 148 117 156 108 168 118 148 113 214 203 160 105 193 137 218 149 186 144 184 144 187 147 189 146 174 132 10 M 116 F 90 139 109 148 107 164 114 163 121 221 193 201 159 199 134 216 144 191 150 192 151 200 152 199 149 181 136 100 M 116 72 163 185 231 165 189 198 185 197 189 189 121 F 89 141 98 129 U 9 190 146 146 145 139 137 146 193 187 152 = a-vtid , 1 I CD n --i m t/i SSE * Dietary Level of A ro clo r 1254 1npr.il Sex Coni rol M F 1M F 10 M F 100 M F T A B LE III continued TEST MATERIAL: A ro clo r 1254 T w o-Y ear Chronic O ral Toxicity Study - Albino Rata Food Consumption Data 45 196 ' 189 148 154 189 IBI 141 159 190 191 139 138 179 168 148 141 Sum m ary of Mean Values Food Consumption (gram s/rat/seven days) Mouth; 6 7 8 9 10 168 147 163 184 179 136 121 138 152 160 11 170 141 184 198 190 190 197 152 147 162 145 150 151 123 189 196 190 188 191 186 148 148 150 145 138 142 190 179 187 182 197 180 144 162 163 151 148 141 12 187 127 190 126 163 149 188 137 . Averago Food C onsum ption (gram s/m l/seven daval 176 142 186 145 187 144 183 148 (. ^ aoh o-03* C* M o r t a l i t y and R e a c tio n s A frequency distribution of natural deaths o ccu rrin g during the in v e s tig a tio n a p p e a rs in T able IV, No untow ard behavioral reactions w ere noted am ong any of the anim als employed in the investigation. ACM OC35 OC 362 Dietary Level (ppm) Sex Control M F 1M F 10 M F 100 M F ,T A B L E IV TE ST M A TER IA L: A ro c lo r 1254 Tw o-Y ear Chronic O ral Toxicity Study * Albino Rate M ortality Data Frequency Distribution of Deaths Frequency D istribution of Deaths M onths: 1-3 4-6 7-9 10-12 13-15 16-1 a 19-21 000 2 17 6 000 1 3 5 7 000 2 5 6 8 20 0 1 2 15 0 O' 0 1 3 5 8 0 11 4 2 3 4 00 1 3 2 5 7 I0 2 4 4 2 4 22-24 9 3 9 3 10 7 9 6 I T otal Dead 1 Number T ested J % 25/35 19/35 30/35 14/35 27/35 22/35 27/35 23/35 Does not Include the five anim als of each sex from every group sacrificed for in terim pathologic stu d ies. ACH G035 CIO u S I /< 13 D. H em atologic and C linical Blood C h em istry Studies M ean m ale and female Hematologic and clinical blood c h em istry d a t a a r e s u m m a r i z e d in T a b le s V th ro u g h DC. Values for all param eters investigated w ere within the norm al ran g e for the albino ra t. No signifieant differences betw een te st and c o n tro l values were observed. . ACM C 0 3 5 6 00 364 OO O D ietary Level (pnm) Control TABLE V TE ST MATERIAL-, A ro clo r 1254 T w o-Y ear C hronic O ral Toxicity Study - Albino Rata Hem atologic Data Sum m ary of Mean Value Sex 3 M 14.0 F 9*1 Total Leukocyte Count (thousands/mnrv*) Months 6 9 12 18 15.4 10. 1 1!. 1 9 .9 9.4 6.7 7.6 6.8 24 11.2 7.6 Erythrocyto Count (m illiona/m m *) Month: 3 6 9 12 IB 8. 1 8 .2 8 .7 8 .8 7 .6 7.5 7.7 7.3 . 7.8 7.7 24 6.5 6.8 M 15.1 15.2 1 0 . 9 91 1 . 0 9.8 10.2 7.5 7.7 8. 1 7.7 8.0 8.0 100 F 11.6 12.2 7 . 3 8 . 7 6 . 6 11.8 7 . 5 7 . 6 7 . 7 6 . 9 7 . 3 6 . 4 O O >o uoU>^1 X o 0 01 o- O TA B LE VI TEST MATERIAL: A ro clo r 1254 T w o-Y ear C hronic O ral Toxicity Study - Albino. Rata H em atologic Data Sum m ary of Mean Valuea D ietary Level (onm) Sex Hemoglobin Concentration (g/100 mil Month: 3 6 9 12 IB 24 3 M 17. 1 1 7 .7 1 5 .7 1 7 .6 16. 1 1 3 .6 51 Control F 1 6 .9 17.4 14. 1 16.7 16.4 14.6 50 H em atocrit Value (V.) 1M o n t h : 6 9 12 IB 49 48 50 41 47 43 . 46 42 24 35 38 M 15.6 17.6 13.8 15.6 15.7 15.3 47 47 42 44 41 40 100 F 16.5 17.8 13.9 14.5 14.5 12.7 48 47 42 40 37 32 O ox LJ ach\ ooIP O' * 9S 00 OO O D ietary Level (pnm) C o n tro l TA BLE VII TEST MATERIAL: A ro clo r 1254 . T w o-Y ear Chronic O ral Toxicity Study - Albino R ats Hematologic Data Sum m ary of Mean Values Sex 3 M 82.0 t F 81.0 D ifferential Leukocyte Count (Number of Celia per Hundred) L ym phocytes N eutrophils Month: Month: 6 9 12 18 24 3 6 9 12 18 88.6 80.4 78.8 79.0 65.4 16.0 11.4 18.6 20.6 20. 0 86.6 86.8 89.0 70.2 61.4 15.4 11.6 11.8 . 10.0 29.2 24 32. 2 36.0. M 88.2 79. A 85.8 85.2 69.8 68.0 10.8 20.2 l i . 8 14.2 29.0 30.2 100 F 92.2 92.0 79.6 84.6 79.0 6l.O 7.4 7.8 20.2 15.0 20.4 57.6 JtiOiU I ! ( \ O I o TABLE Vll continued TEST MATERIALS A roclor 1254 T w o-Y ear Chronic O ral Toxicity Study - Albino Rata Hem atologic Data Sum m ary of Mean Values D ietary Level (npm) Control Sex 3 M 0.6 F 0.6 D ifferential Leukocyte Count (Numbor of Cell a p er Hundred) M onocytes Eosinophils Month: Month: 6 9 12 18 24 3 6 9 12 18 0 .0 0.0 0.4 0 .2 1.2 1.4 2.0 1.0 0 .2 0. 8 0 .0 0.0 0.0 0 .2 1.0 1.0 1.8 1.4 0 .2 0 .6 24 1.6 1.6 M 0 .0 0 .0 0.0 0 .0 0.0 0.8 1.0 0 .2 0.4 0.6 1.2 1.0 100 F 0.0 0.0 0.0 0.0 0.0 1.0 0.4 0.2 0.2 0.4 0.6 0 .4 AON 0 C 3 5 6 5 OO D ietary Level (ppm) C o n tro l Sex M a F M 100 F TADLE VII continued TEST M ATERIAL: A roclor 1254 T w o-Y ear Chronic O ral Toxicity Study - Albino Rata Hem atologic Data S um m ary of Mean Value Differential Leukocyte Count (Number of Cella por Hundred) Basophile Month: 3 6 9 12 IB 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 24 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 o AOM 003 *o r i c a I I (l T A B L E VIII T E ST M ATERIAL: A ro c lo r 1254 T w o-Y ear C hronic O ral Toxicity Study - Albino Rata Hematologic Data Sum m ary of Mean Values D ietary Level (ppm) Sex 3 Serum Glutam ic-Pyruvic T ransam inase Activity (Dado Units) Month: 6 9 12 18 24 3 Blood U rea Nitrogen Concentration (m g s */) Month: 6 9 12 18 M 38 48 45 45 42 36 17 15 12 10 14 Control F 33 39 38 42 32 30 14 16 15 9 13 M 33 32 43 ' 50 38 41 14 15 13 12 11 100 F 32 46 49 61 46 78 16 15 13 14 13 24 15 11 15 15 O C3 ACM OC 3 5 67 O OO D ietary Level (ppm) Control Sex M F TA B LE IX T E S T M ATERIAL: A ro c lo r 1254 T w o-Y ear Chronic O ral Toxicity Study - Albino Rata Hematologic Data Sum m ary of Mean Values F asted Blood Glucoeo Serum Alkaline Phosphatase Concentration Activity (m g s 7.) (K lng-A rm strong Units) Mouth: Month: 3 6 9 12 18 24 3 6 9 12 18 24 128 123 89 n o n a 115 88 138 99 28.6 24.7 21.5 20.3 16.2 22. 3 92 134 109 18.9 1 7.8 10.6 11-9 1 6 .8 2 0 .2 M 109 113 129 128 124 115 3 5 .4 2 3 .2 2 7 .4 27. 1 1 4 .6 2 3 .5 100 F 120 107 124 113 132 107 17. 1 18. 1 1 9 .3 1 6 .7 1 0 .0 2 1 . 6 r ( i u a ACM 0 0 3 5 6 S fat jiv L lU 'm iL B 10 T S S T a S y ia t& w i. $*%> Zh E* U rin e A nalyses Me^n m ile and fem ale urine analyses data are sum m arised in Tables X through XU. No differences w ere noted between the urine from control and te st anim als at the intervals of examination. ADM C 0 3 5 6 9 * 0 0 372 I TABLE X TEST MATERIAL: A ro clo r 1254 T w o-Y ear Chronic O ral Toxicity Study - Albino Rata Urine Analyses Data Sum m ary of Mean Values D ietary Glucose Albumin Level Month: Month: (ppm) Sex 3 6 9 12 18 24 3 6 9 12 i a 24 Control M F nn n *n n n n n nn n t n *n n n n n t 42 43 nt 42 Mn n n n n nn n t t t 42 100 a Fn n n n n nn n t tt 43 O> nrt ~K U) NJ ;> Co O It) Key: n - negative t = tra c e ; le s s than 30 m g /1 0 0 m l u rin e 4 1 = 30 to 100 m g / 100 m l u r i n e -t-2 - 100 to 300 m g /1 0 0 m l u r i n e 3 = 300 to 500 m g / 100 m l u r in e 4*1 = m o r e than 500 m g /1 0 0 m l u r i n e Ui r. Oc o BI0 -TE3T T A B L E XI TEST M ATERIAL; A ro cto r 1254 T w o-Y ear Chronic O ral Toxicity Study - Albino Rata U rine Analyses Data Sum m ary ol Mean Values Di i-i a r y M icroscopic Elements pH Level Month: Month: !P* S*x 3 69 12 18 24 1 6 9 12 IB 24 a # * % *s n n C r. * n n n n 11 7 7 6 nt t887 77 86 6 6 n n S s nt t776 76 6 t1 p j* n n N t t 887 76 6 Key: \* - N o r m a l n a negative O I = trace; less than 5 per low power Held o 41 = 5 lo 10 p e r low p o w o r field -I 2 = 10 to 15 p e r low p o w e r fie ld 11 = m o r e th a n 15 p e r low p o w e r fie ld s e o o huv * V. D ietary Level (ppm) Control Sex M F M 100 F TABLE XII T E ST M ATERIAL: A ro clo r 1254 T w o-Y ear Chronic O ral Toxicity Study - Albino Rats U rine Analyses Data 3 1*029 1.020 Sum m ary of Mean Values Specific Gravity Month: 6 9 12 1.022 1.033 1.027 1.019 1.025 1.024 18 1.042 1.032 1.01B 1.020 1.021 1.018 1.029 1.023 1.027 1.022 1.037 1.029 24 1.05) 1.039 1.027 1.031 V* ta O n U) t K Zl)> tUU WOv se*i JtVl*1V* F . Pathologie Studies 1. T hrceM onth Sacrifice I G ross Pathologie Findings No outstanding differences w ere noted between test and control rats upon gross pathological exam ination. b. O rgan "eight and Organ to Body and O rg a n to 5 r a i n V.'oigiit R atio D ata The results of the statistic a l analyses conducted on absolute org an w eights, organ to body w eight and o rg an to b rain weight ra tio s a re s u m m a riz e d in Tables XIII th rough XVIII. Significant differences between a te st group and the control group a re designated by asterisks following the te st value. O rgan w eights, organ to body w eight and organ to b ra in weight ratios disclosed several randomly occurring intergroup differences. The lack of any consistent dose related response and the absence of any deleterious histopathologic change confirm that these differences were not related to the ingestion of A roclor 54. AOM 0 0 3 5 7 3 OC 376. o TABLE X ia T E ST M ATERIAL: A ro c lo r 1254 T w o-Y ear Chronic O ral Toxicity Study - Albino Rat* O rgan Weight and Ratio Data Three-M onth Sacrifice Sum m ary of Mean Values Organ: Liver Dietary Level of A ro c lo r 1251 (ppm) O rgan Weight O rgan/B ody Weight Ratio (g) (g /1 0 0 g) M a le s _______F e m a l e * __________ M a le a _______F e m a l e * O rg an /B rain Weight Ratio (g/g) M a le s _______F e m a l e * C o n tro l 1 10 100 o 21.4 24.0 22. B 28.0* 13.0 13.5 13.1 13.8 4.13 4.34 4.59 5.25 4.16 4.34 4.29 4.89 10.3 11*7 10.9 14.6** 6.82 6.85 6.85 7 . 11 S tatistically significant difference at the 95 p e rc en t confidence level. ** S ta tistic a lly significant d ifferen ce at the 99 p e rc e n t confidence lev el. .seoo uo/ B l 0 - T E 5 T ACM OC 0 0 378 D iiU ry Level of A ro c lo r 1254 (pnml C o n tro l 1 10 100 TA B LE XIV T E ST MATERIAL! A ro c lo r 1254 T w o-Y ear Chronic O ral Toxicity Study - Albino Rats Organ Weight and Ratio Data Three-Month Sacrifice Sum m ary of Mean Values Organ: Kidneys * O rgan W eight (6) M ales Fem ales 4.03 4.30 4.05 4.21 2.49 2.62 2.61 2.74 O rgan/B ody Weight Ratio (g /1 0 0 g) M ales Fem ales 0.782 0.802 0.778 0.845 . 0.B21 0.792 0.852 0.975 O rgan/B raln W eight Ratio (g/g) M ales Fem ales .1 .9 5 1.31 2.09 1.32 1.94 2.19 1.36 1.42 u* u oo o t a b Le XV TE ST M ATERIAL: A ro c lo r 1254 T w o-Y ear Chronic O ral T oxicity Study - Albino R ats Organ Weight and Ratio Data Three-M onth Sacrifice Sum m ary of Mean Values Organ: Spleen Dietary Level of O rgan Weight O rgan/B ody Weight Ratio O rg an /B raln Weight R atio A r o c lo r 1254 (g) (g /1 0 0 g) (g/g) fpnm)______________M a le s ______ F e m a l e s __________ M a le s _______F e m a l e s __________ M a le s ______ F e m a l e s s C o n tro l 0.766 0.578 0.150 0.184 0.371 0. 302 1 0.942 0.668 0 . 167 0.217 0.454 0.338 10 0.766 0.586 0.154 0.191 0.368 0.308 100 0.814 0.542 0.153 0.192 0.425 0.281 AOM C 03576 n/m- r c Dietary Level of A ro c lo r 1254 (PPm) _ _ C o n tro l 1 10 100 TA B L E XVI T E ST M ATERIAL: A ro c lo r 1254 T w o -Y ear Chronic O ral Toxiclly Study - Albino Rale Organ Weight and Ratio Data Three-M onth Sacrifice . Sum m ary of Mean Values Organ; Gonads O rgan W eight (g) M ales Fem ales 3.60 0.114 3.50 0.134 3.55 0.110 3.82 0.120 O rgan/B ody W eight Ratio (g /1 0 0 g) M ales Fem ales 0.698 0.0370 0.637 0.0431 0.720 0.0361 0.718 0.0426 O rgan/B rain Weight Ratio fg/g) M ales Fem ales 1.74 1.70 0.0599 0.0675 1.70 0.0568 1.99 0.0618 i 3 I > r oo o TA B LE XVII TE ST M ATERIAL: A ro c lo r 1254 T w o-Y ear Chronic O ral Toxicity Study - Albino Rata Organ Weight and Ratio Data Three-M onth Sacrifice Sum m ary of Mean Value Organ; Heart Dietary Level of O rgan Weight O rgan/B ody Weight Ratio O rgan/B raln W eight R atio A ro c lo r 1254 (g) ( g /1 0 0 g) (g/g) (ppm )______________M ale______ F e m a l e __________ M a le _______F e m a l e ._________ M ale ' F e m a l e C o n tro l 1 10 100 1.59 1.56 1.49 1.58 1.05 1.02 0.984 0.992 0.307 0.283 0.302 0.297 0.339 0.332 0.322 0.352 0.767 0.756 0.713 0.817 0.551 0.516 0.515 0.512 ADM . 0 0 3 5 7 8 " fctSEOO rx Dietary Level of A ro c lo r 1254 (ppm) C o n tro l 1 O >n x ou>o 1 100 r O TABLE XVII! T E ST M ATERIAL: A ro c lo r 1254 Tw o-Y ear Chronic O ral Toxicity Study - Albino Rata O rgan Weight and Ratio Data Three-M onth Sacrifice Sum m ary of Mean Values Organ; Drain O rgan Weight (g) M ales Fem ales 2.07 1.91 O rgan/Body Weight Ratio (g /1 0 0 g) M ales Fem ales 0.402 0.616 Final Body W eights *f Sacrificed Rata (g) M ales Fem ales 518 314 2.07 1.99 0.376 0.647 553 311 2.09 1.91 0.424 0.625 * 499* 307 1.95 1.94 0.366 0.668 434 261 BIC .'EST i: C. H is to p a th o lo g ic T in d in g s H istopathologic exam ination of tissu e s and organs ta k e n f r o m a ll a n im a ls fro m both the c o n tro l and 100 ppm g ro u p s w as conducted. T a b le s XIX and XX lis t a ll histo p ath o lo g ic changes noted. A ll of the lesions noted in the m ic ro s c o p ic ex am in a tion of tis s u e s w ere those of spontaneous d isea se and a re not unusual for the albino rat. The m ost frequent findings w ere lesions in the tra c h e a and lungs* indicating chronic m urine pneum onia. T hese oc c u rre d in the control as well as the rats fed A roclor 1254. AOM 0 0 3 5 SC nn 383 T A B L E XIX T E S T M A TER IA L; A ro c lo r 1254 T w o-Y ear Chronic O ral Toxicity Study - Albino Rats Histopathologic Changes c Three-Month Sacrifice Croup: Control N um ber of A nim als Organ Examined Findings A vere Incidence C rac 5 Males Trachea Lung U rinary Bladder Tracheitis Chronic resp irato ry disease H yp erp lasia 1 1.0 1 * 1.0 3 1.0 1 5 Fem ales Lung Chronic respiratory disease 1 1.0 AU other tissues and organs w ere norm al histologically. Grading System 0.5 = m inim al 1.0 = slight 2 . 0 = mild 3.0 = m oderate 4.0 = severe 5.0 = extrem e A DM 0 0 3 5 . 0 ( 384 ) T A B L E XX T E S T M ATERIA L: A ro c lo r 1254 Tw o-Y ear Chronic O ral Toxicity Study - Albino Rata Histopathologic Changes Three-Month Sacrifice G ro u p : 100 p p m N u m b e r ox Average A n i m a l s - O r g a n E x a m i n e d - ---------- F in d in g s _____ . . In c id e n c e _ ...G rad e 5 Males Trachea Lung Tracheitis Chronic resp irato ry disease 1 * 3 1.0 1.0 5 Fem ales Trachea Tracheitis 1 1.0 All other tissues and organs were norm al histologically. Grading System 0.5 = minimal 1.0 = slight 2 .0 = mild 3 . 0 = moderate 4 . 0 = severe 5.0 = extreme % ACH 00 3 5 8 2 OC 385 ,W r -'^ a I C 3 I SMZ&UCl. ,u 2. 5is-Mnrth ^acrifico a. G ross Pathologic Findings No outstanding differences w ere noted between te st and control anim als. b. O rgan Weight ar.d O r? m to Bodv and O rg a n to B ra in V.'eigh; Ratio D ata The re su lts of the statistical an aly ses conducted on absolute' organ weights , organ to*body weight and organ to brain weight " ra tio s a re su m m arized in T ables XXI through XXVL O rgan w eights, organ to body w eight and organ to b ra in weight ratios disclosed several randomly occurring intergroup differences. The lack of any consistent dose related response and the absence of any deleterious histopathologic change confirm that these differences w ere not re la te d to the ingestion of A ro clo r 1254. ADh CC35a n r t a fs 1w * - J 1 9 Dietary Level of A ro c lo r 1254 C o n tro l 1 10 100 TA BLE XXII i T E ST M ATERIAL: A ro c lo r 1254 T w o-Y ear Chronic O ral Toxicity Study - Albino Rata Organ Weight and Ratio Data Six-Month Sacrifice Sum m ary of Mean Values Organ: Kidneys O rgan Weight (b) M ales Fem ales 4.04 2.79 3.80 2.58 3.22** 2.50 3.23'* 2.43 O rgan/B ody Weight Ratio ( /1 0 0 g) M ales Fem ales 0.664 0.806 0.715 0.784 0.703 0.780 0.700 0.861 O rgan/B raln W eight Ratio (g/l M ales Fem ales 2.06 1.43 1.75* 1.39 1.57* . 1.31 1.68** 1.25 * Statistically significant difference a t the 95 p e rc en t confidence level. .* t S 5 0 0 WOV riM 0 0 3 5 6 TARLE XXI T E ST M ATERIA L; A r o d o r 1254 * T w o-Y ear C hronic O ral Toxicity Study - Albino Rata Organ Weight and Ratio Data % Six-Month Sacrifice Sum m ary of Mean Valuea Organ: Liver Dietary Level of O rgan Weight O rgan/D ody Weight Ratio O rg an /B raln Weight Ratio A ro c lo r 1254 (g) (g /1 0 0 g) <g/g) (pn:n)______________M ale_______F e m a l e __________ M a lc a ______ F e m a l e s __________ M a le a Fem ales C o n tro l 22.9 13.1 3.74 3.77 11.7 6.71 1 16.1* 10.3* . 3 . 0 f* `* 3.11** 7.45** 5.54* 10 12. 9 .5 4 * 2.81** 2.94** 6.29** 4 .9 9 * 100 19.2 12.4 4.14 4.40 10.0 6.39 * S tatistically significant difference at the 95 p e rc en t confidence level. S ta tis tic a l)-/ s ig n ific a n t d if f e r e n c e a t the 99 p* e r c e n t c o>n fid e n ce le v e l. i L ( ft I; II. tt- i; t K O D.clary Level of A ro c lo r 1254 fopm) Control 1 10 100 O 4 TABjLE XXIII TE ST M ATERIAL: A ro c lo r 1254 Tw o-Y ear Chronic Oral Toxicily Study - Albino Rats Organ Weight and Ratio Data Six-Month Sacrifice Summary of Mean Values Organ: Spleen e o Organ Weight <6> Males Females 0.752 0.684 0.830 0.602 0.588 0.594 0.664 0.512 Organ/Dody Weight Ratio (g/ioo e) Males Females 0.122 0.196 6.155 0.182 0.128 0.184 0.150 0.180 O rgan/B raln Weight Ratio (c/c) Males F em ales 0.384 0. 350 0.384 0.324 0.287 0.311 0.356 0.264 ACM 0 0 3 5 3 6 I AC 0 0 J 5 S 7 o % Dietary Level of A ro c lo r 1254 (onml Cont rol 1 10 100 Oo T A B L E XXIV T E S T M ATERIAL: A ro c lo r 1254 Tw o-Y ear Chronic Oral Toxicity Study - Albino Rata Organ Weight and Ratio Data Six-Month Sacrifice Sum m ary of Mean Values Organ; Gonads a Organ Weight (g) Males F em ales 3.55 0.0960 3.67 0.0922 3.30 .0.0868 3.44 0.104 Organ/Body Weight Ratio ( g / 1 0 0 g) Males Females 0.593 0*. 691 0.0279 0.0280 0.724 0.0272 0.747 0.0364 O rgan/B ratn Weight Ratio (g/g) Males Fem ales 1.81 0.0492 1.70 0.0496 1.61 0.0454 1.79 0.0536 | ! i ADH CC3 5 a Dietary Level of A ro c lo r 1254 (ppm) Control l 10 100 TA BLE XXV / TEST M ATERIAL: A ro c lo r 1254 . - Two-Year Chronic Oral Toxicity Study - Albino Rata Organ Weight and Ratio Data Six-Month Sacrifice ,, Summ ary of Mean Valuoa Organ: Heart a , Organ Weight <8> 'Malea Femalea 1.70 1.05 1.58 1.04 1.44 1.03 1.40 1.02 Organ/Body Weight Ratio ( g / i o o g) M ale. Femalea 0.282 0.302 0.298 0.316 0.314 0.320 0.301 0.361 O rgan/B rain Weight Ratio (g/fi) Malea F em ales 0.867 0.538 0.731 0.559 0.702 0.539 0.729 0.526 O Dietary Level of A ro c lo r 1254 (pnm) Control 1 10 100 oQ T A*B LtE XXVI TE ST MATERIAL* A ro c lo r 1254 Two-Year Chronic Oral Toxicity Study - Albino Rata Organ Weight and Ratio Data Stx-Monlh Sacrifice Summary of Mean Values Organs Drain e - Organ Weight , Organ/Body Weight Ratio (g) (g /1 0 0 g) M a l e s ______ F e m a l e s __________ M a l e s ______ F e m a l e s Final Body Weights of Sacrificed Rats (g) M a l e s _______F e m a l e s 1.96 2.16 1.95 1.86 0.320 0.4044 0.560 0.562 613 348 534 331 2.05 1.92 1.91 1.94 0.447>* 0.414** 0.593 0.681*' 459 322 464 285 t Statistically significant difference at the 95 p e rc en t confidence level. Statistically significant difference at the 99 percent confidence level. 392 \mjy c. Histopathologic Flndlr.es H istopathologic exam ination of tissu e s and organs ta k e n fro m a ll a n im a ls fro m both th e c o n tro l and 100 ppm g ro u p s w as conducted. T ables XXVII and XXVffi list all histopathologic changes noted. All of the lesions noted in the m icro sco p ic exam ination of tis su e s w e re those of spontaneous d isease and a r e not unusual fo r the albino ra t. The m o st frequent findings w ere lesions in the trach ea and lungs, indicating chronic murine pneumonia. These occurred in the c o n t r o l a s w e l l a s th e r a t s fed A r o c l o r 1254.03 *G*V 0 0 3 S ,C 393 I 0 _J TA B LE XXVII TEST M ATERIAL; A ro clo r U S4 T w o -Y ear C hronic O ra l Toxicity Study - A lbino Rata H istopathologic Changes Six-M onth S acrifice C roup: C ontrol N u m b e r o Animals Orcan Examined Findings 5 M ales . T rachea Lung L iver S k e le ta l M uscle T racheitis Focal tracheitis F o cal pneum onitis H yperem ia F ocal pericholangitis Fo cal inflam m ation 5 Fem ales T raehea Lung L iver K idney Focal tracheitis H yperem ia Hype rem ia P ericholangitis F ocal inflam m ation Averag Incidence G rade 2 1.5 2 1.0 1 * 0.5 2 1.0 * 1 1.0 I 1.0 1 2' 1 1 1 1.0 2 .5 1.0 0 .5 1.0 A ll other tissu es and organs w ere norm al h isto lo g ically . G rading System 0 .5 * m inim al 1 .0 s slight 2 .0 m ild 3 . 0 * m oderate 4 .0 severe 5 .0 * extrem e 003591 *0* tablexxvm T E S T M A T ER IA L : A r o c l o r 1254 ^ v o -Y e a r Chronic O ral Toxicity Study - Albino Rats Histopathologic Changes J Six-Month Sacrifice G ro u p : 100 pp m N u m b e r of Animals Organ Examined Findings Ave rage Incidence G rade 5 Males Trachea. Lung Liver ' Kidney 5 Females H eart Trachea Lung Kidney Tracheitis Focal tracheitis Focal pneumonitis Focal pericholangitis Focal nephritis - Myocarditis Foeal tracheitis Focal pneumonitis Hyperemia * Focal nephritis 2 1 3 1 *l 1 3 .. 4* 1 2 2.0 1.0 0.5 2.0 0.5 0.5 0.5 1.0 1.0 3.0 All other tissues and organs were normal histologically. Grading Svrstam 0.5 s minimal 1.0 = slight 2.0 a mild 3.0 s moderate 4.0 ssevere 5.0 * extreme ADM 0 0 3 5 9 2 395 01 BIO TES T 3* T w e l v e Month S a c r i f i c e a* G r o s s P a t h o l o g i c Fin d i n g s Cross pathologic findings among test animals were not significantly different from those noted in control anim als, b. O r g a n V.rci g h t and R at io D a ta The mean organ weight and ratio data collected at the 1 2 - m o n t h s a c r i f i c e a r e p r e s e n t e d in T a b l e s XXDC t h r o u g h XXXIV. Statistically significant differences, are designated by asterisks following the test group value. Organ weights, organ to body weight and organ to b ra in weight ratios disclosed several randomly occurring intergroup differences. The lack of any consistent dose related response and the absence of any deleterious histopathologic change confirm that these differences were not related to the ingestion of A roclor 1254. *0 * 0 J 593 Of 3 9 6 r Dietary Level Inpnl Coni rol 1 10 100 TABLE XXIX TEST MATERIAL; A ro clo r 1254 Tw o-Y ear Chronic O ral Toxicity Study - Albino Rate Twelve-Month Sacrifice Organ Weight and Ratio Data Sum m ary of Mean Values Organ Weight (g) Males Females 22. 1 15.5 20.6 12.9 23.5 14.6 27. 1 12.2 Organ: Liver . Organ/Botly Weight Ratio (g /1 0 0 g) Males Females 3.79 3.52 3.09* 3.41 3.52 4.34 4.58* 3.43 O rgan/Braln Weight Ratio (g/g) Males F em ales 10.8 7.72 9.89 7.29 11.2 7.3d 12.7 A. 30 * Statistically significant difference at (he 95 p e rc e n t confidence level. AC u u> k Dietary Level (oaml Control 1 10 100 TA B LE XXX TEST MATERIAL: A ro clo r 1254 Tw o-Y ear Chronic O ral Toxicily Study - Albino Rats Twelve-Month Sacrifice Organ Weight and Ratio Data Summary of Mean Values Organi Kidneys Organ Weight (g) Males Fem ales Organ/Dody Weight Ratio (g/100 g) Males Females O rgan/Braln Weight Ratio (g/g) Males F emalcs 4.04 4.09 4.64 4.39 3.19 2.75 2.76* 2.78* 0.692 0.618 0.697 0.740 0.738 0.728 0.817 0.796 1.97 1.98 2.21 2. 06 1. SB 1.55 1.39 1.44 O ct uu>> oo * Statistically significant difference at the 95 p ercen t confidence level. i r Dietary Level (oom) Control 1 10 100 TA BLE XXXI TEST MATERIAL: A ro c lo r 1254 Tw o-Y ear Chronic O ral Toxicity Study - Albino Rata Twelve-Month Sacrifice . Organ Weight and Ratio Data Sum m ary of Mean Valuea Organ; Spleen Organ Weight (g) Males Females 0.778 0.714 0.904 0.796 0.4B4+ 0.548 0.754 0. 976 Organ/Body Weight Ratio (g /1 0 0 g) Males Females 0. 133 0.183 0.107 0.129 0.135 0.161 0. 126 0. 280 O rgan/Braln Weight Ratio fg/g) Males F em ales 0. 380 0.196 * 0.344 0. 271 0.430 0.275 0.352 0.517 ti v. C3 O in in y. V n * S ta tis tic a lly significant difference at the 95 p e r c e n t confidence le v e l.' IP U) IP 399 r 400 Dietary Level (nnm) Control 1 10 too o o o o UUJl TA B LE XXXII TE ST M ATERIAL: A ro clo r 1254 Tw o-Y ear Chronic O ral Toxicity Study - Albino Rats Twelve-Month Sacrifice Organ Weight and Ratio Data Summ ary of Mean Values m Organ: Gonads Organ Weight (g) Males Females O rgan/nody Weight Ratio (g /1 0 0 g) Males Females 3.46 0.116 0.592 0.0275 3.68 0. I l l 0.557 0.0294 3.68 0.0918 0.550 0.0270 3.56 0. 121 0.603 0.0334 Organ/Brain Weight Ratio (g/gj Males F em ales 1.70 0.0574 1.78 0.0624 1.75 0.0461 1.67 0.0624 V E, cr I < i L r(t ir- * fi >1 in 4- O TA B LE XXXIII o TEST MATERIAL; A roclor 1254 Tw o-Y ear Chronic O ral Toxicity Study - Albino Rats Twelve-Month Sacrifice Organ Weight and Ratio Data Sum m ary of Mean Values Organ; Heart Dietary Level (opm| Contro) 1 0 100 Organ Weight Males (8) Females 1.71 1* 23 1.68 1*13 1.84 1.08** 1.69 1*14 Organ/Body Weight Ratio (k/ ioo b) Males Females 0.293 0. 286 0.255 0.301 0.275 0.322 0.2B4 0.325 Organ/Brain Weight Ratio (/> Males F em ales 0.835 0.609 0.812 0.635 0.875 0.544* 0.795 0.589 I /<(C, ACM 0 0 3 5 C * S ta tistic a lly significant d ifferen ce at the 95 p e r c e n t confidence level* *> S t a t i s t i c a l l y s i g n i f i c a n t d i f f e r e n c e at th e 99 p e r c e n t c o n f i d e n c e l e v e l . ooo Dietary Level fpnml Control I 10 too TA B LE XXXIV TE ST MATERIAL: A roclor 1254 Two-Year Chronic Oral Toxicity Study - Albino Rate Twelve-Month Sacrifice Organ Weight and Ratio Data Summary of Mean Valuee Orga n Weight <g> Males Females 2.04 2.02 2.08 . 1.78 Organ: Brain Organ/Body Weight Ratio <6/100 6) Males Female 0.35 0.474 0.315 0.475 Final Body Weight of Sacrificed Animala (grama) Males Females 584 4 39 665 360 2. 10 2. 14 1.99 1.93 0. 315 0.361 0.591 0.552 669* 596 . 338* 355 ' S t a t i s t i c a l l y significant difference at the 95 p e r c e n t confidence level. O >D o3 K> OOiiPn u O c Histopathologic Findings Histopathologic examination of tissues taken from each rat s a c r i f i c e d f r o m th e c o n t r o l and 1 0 0 'ppm g r o u p s a f t e r 12 m o nth s of t e s t i n g was conducted. T a b l e s XXXV and XXXVX l i s t a l l h i s t o p a t h o l o g i c c h a n g e s noted* AO* 0C3tCC A^taJL B I O " E S T ^ 30 TA BLE XXXV TEST MATERIAL: A roclor 1254 T w o-Y ear Chronic Oral Toxicity Study - Albino Rate Histopathologic Changes Twelve-Month Sacrifice Group: Control N u m b e r of Animals 5 Mile a 5 Females - Organ Examined Trachea Lung Urinary Bladder Trachea Lung Colon Findings Tracheitis Chronic respiratory disease Calculus Trache itis Chronic respiratory disease Parasites Incidence 1 2 1 i.. 1 1 Aver Cra 1.1 1.1 . l.< 1.1 1.1 1.1 All other tissues and organs were normal histologically. Grading System 0.5 = minimal 1.0 = slight 2.0 = mild 3.0 =.m oderate 4.0 = severe 5.0 = extrema ADh 0C36C1 0 C 44 jfjCY LiAiLal 8 I C T E S T j Uc t'i TABLE XXXVI TEST MATERIAL: A ro c lo r 1254 Two-Year Chronic Oral Toxicity Study - Albino Rats Histopathologic Changes Twelve Month Sacrifice Croup: 100 p p m N um ber of Organ Animals Examined Findings Averi - - Incidence * "G rad: 5 Males Lung Chronic respiratory disease 4 1.0 5 Females Heart Trachea Lung Adrenal Focal myocarditis Tracheitis Chronic respiratory disease * Congestion i 1 1 1 1 1.0 1.0 1.0 1.0 AH other tissues and organs were normal histologically. Gradir.s Svs tern 0. 5 * m inimal 1. 0 a slight 2. 0 = mild 3. 0 = m oderate 4. 0 = severe 5.0 s extreme ADM 0036C 405 00 VO I W - T 5 5 I i. 60 i 4. F i n a l Sacrifice - a*. C r o s s P a th o lo g ic Findings i\ [ i G ross pathologic findings among test animals were not significantly different from those noted in control anim als, b. Organ height and Ratio Data The mean organ weight and ratio data collected at the final sacrifice are presented in Tables XXXVE through XLII. Statistically significant differences, a r e designated by asterisk s following the test group value. Absolute liver weight, liver to body weight and liv e r to b ra in weight ratios were significantly elevated in both male and female r a t s fed 100 p p m A r o c l o r 1254. O t h e r o rg a n w e i g h t s , o r g a n to b o d y weight and organ to brain weight ratios disclosed sev e ra l randomly oc curring intergroup differences. The lack of any consistent dose related r e s p o n s e and the absence of any deleterious histopathologic change confirm that these differences w ere not related to the ingestion of A roclor 1254. AOM 0 C 3 t 0 0 0 0 406 o 0 0 0 OO Dietary Level . (pprni Control 1 10 100 TABLE XXXVII T E ST MATERIALS A ro c lo r 1254 Two-Year Chronic Oral Toxicity Study - Albino Rata Organ Weight and Ratio Data i Sum m ary of Mean Values Final Sacrifice Organ Weight Males <el Females Organ: Liver Organ/Body Weight Ratio ( s / > K) Males Females 22.0 16.3 3.47 3. OS 19.5 16. 1 2.99 3. 10 23.9 19.5 3.21 3.84 3.0, 2** 3 2 . I+* 4 . 75* 8.62** * Statistically significant difference at the 95 percent confidence level, * ** Statistically significant difference a t the 99 p e rc e n t confidence level. AUh 003604 o 0 i, t ii;' O r g a n / B r a l n W e i g h t R a t i o ** (o/l Males Females 10, 10 8.42 8.76* 8. 31 po^i o* 1 1 . 40 1 4 . 10* 17. 10* O' OO Dietary Level (ppm) Control 1 10 100 TABLE XXXVIII T E S T M A T ER IA L : A ro c lo r 1254 Two-Year Chronic Oral Toxicity Study - Albino Rats Organ Weight and Ratio Data Sum m ary of Mean Values Final Sacrifice Organ Weight Males lei Females Orgahi Kidneys Orgau/11 tidy Weight Ratio lit 100 g) Males Females 5.57 3.48 0. 877 0.646 5.52 ) . 68 0.868 0. 706 5.6) 3.47 0. 761 0.717 4.74 3.44 0.749 0.899* Organ /B rain Weight Ratio M ales (e/el Fem ales . 2.57 1.79 2.48 1.8B 2.70 1.84 2.21 <M 09 . ''*S t a t i s t i c a l l y s ig nifi c a nt d i f f e r e n c e a t the 95 p e r c e n t c o n fi d e n c e l e v e l . sot- 0 0 0 AOH 00360*5 a lv r * Dietary Level tppm) Control 1 10 100 TABLE XXXIX TE ST M ATERIAL; A roclor 1254 Tw o-Year Chronic Oral Toxicity Study - Albino Rale Organ Weight and Ratio Data Sum m ary of Mean Values Final Sacrifice Organ Weight Male (Cl Females Organ; Spleen Organ /Body Weight Ratio ( e / i o o g> Males Females 1. IS 0 .9 6 0. 182 0.181 1.26 1 57 1.06 0.86 0. 189 0.220 0.237 0. 175 1.04 0.66 0.164 0. 174 CJ V 1 i Vi O r g a n / B r a l n W e i g h t R a t i o 1 (c/el Males F em ales 0. 520 0.568 0.492 0.562 0.749 . 0.462 0.489 0.352 O O oo X. ou> - oo id Oof>' o Dietary Level (ppmt Control 1 10 100 O . TABLE XL T E ST M ATERIAL: A roclor 1254 Tw o-Y ear Chronic O ral Toxicity Study - Albino Rata Organ Weight and Ratio Data i Summ ary of Mean Values Final Sacrifice Organ Weight Males (el Females 2.52 0.123 Organ: Gonads Organ /Body Weight Ratio (g/100 g) Males Females 0.397 0.0222 3.19 3.28 0.204 0.135 0.482 0.41$ 0.0390 0.0283 . 3.44 0. 128 0.547 0.0316 t ** E o C O r^ a u /B ra in We Ight Ratio (/fil Males F em ales 1. 15 0.0628 1.43 0. 1010 1. 58* 0.0707 1.60 0.0666 Statistically significant difference at the 95 percen t confidence level. noo 410 ADM 0 0 3 6 0 7 oo Dietary Level fpoml Control 1 10 100 TABLE XLI T E ST MATERIAL* Ar o d o r 1254 Tw o-Year Chronic Oral Toxicity Study - Albino Rata Organ Weight and Ratio Data I Summary of Mean Values Final Sacrifice Organ: Heart Organ Weight Males () Females Organ/ilody Weight Ratio (g/100 g) Males Females 1.87 1.54 0.294 0.288 1.98 1.55 0.303 0. 302 2.08 1.37 0.279 0.282 1.94 1.50 0.304 0.386+* * Statistically significant difference at the 95 percent confidence level* `' ^ S t a t i s t i c a l l y s ig n if ic a n t d i f f e r e n c e a t the 99 p e r c e n t c o n fi d e n c e level* O O ^ ADK 0 0 3 6 0 f t If.,, 0 t 1 -4 II tk --4 6 O rg an /B rain We Ight Ratio (/) Males Fem ales 0.860 0. 795 0. 8B9 0.805 0.998* 0.727 0.909 0. 803 * S IO 1 -ST o Dietary Level (ppm) Control 1 10 100 TABLE XLII T E S T M ATERIAL: A ro c lo r 1254 Tw o-Y ear Chronic Oral Toxicity Study - Albino Rata Organ Weight and Ratio Data i Sum m ary of Mean Values , Organ Weight j (s) Males . Females Final Sacrifice Organ: llraln O rgan/Body Weight Ratio (a /lo o g) Males Female Final Body Weight of Sacrificed Animals (grams) Ma les Females . 2. 18 2.22 2.08 . 2. 13 1.95 1.94 ' 1.88 1.88 0. 342 0.347 a 0.279 0. 340 0. 362 0.376 0.388 0.486** 642 551 656 540 762 500 656 411* ** Statistically significant difference at the 99 p e rc e n t confidence level. zx* 0 0 0 ADM 0 0 3 6 0 9 Lm -m L BUT T E S T / a i 67 C* H is to p at h o lo g ic Findi ngs Histopathologic examination of tissues and organs taken f r o m a ll anim als sacrificed a fte r 24 months on te st was conducted. Tables XLItt through XLVI list all histopathologic chang n o t e d . T a b l e XJLVII c o nt ai ns the r e s u l t s of the h i s t o p a t h o l o g i c e x a m i n a t i o r of the urinary bladders. Significant liver injury was found in a n im als fed 100 ppm A roclor 1254 (T-IIfl. The compound associated lesions consisted of vacuolar change, focal hypertrophy and focal hyperplasia. Jn addition, there w ere lesions of inflammation, n ecrosis, fibrosis and m in o r d e generation, w ere more severe and frequent in the T-UI group.. The vacuolar change was not seen in the control anim als b u t w a s o b s e r v e d o c c a s i o n a l l y in l i v e r s f r o m T - I (1 ppm) and T - I I (10 p p m ) anim als. It was frequent in the T-III animals. This lesion is morphologies indicative of fatty degeneration. Form alin-frozen sections of livers fro m representative animals which displayed vacuolar changes were stained with Oil Red 0 to rev eal the presence of fat. The vacuolar lesion in the cytoplas of these cells was positively idenified as fat. I*- 4I The hypertrophic changes found in the liver was focal and often lim ited to the central lobular area where groups of cells were swollen i to two o r th re e times their n o rm a l size with clear pink homogeneous c y to plasm. The hyperplasia was associated with the same cells and appeared AC* 00361C o o c 413 v IW ' I 68 to be i n extension of the hyp ertro p h ic lesio n . The hyperplastic ceils w e r e a ls o u sually h y p ertrophic. The m o s t s e v e r e exam ples of h y p e rp la s ia e appeared as nodular growths with lim ited c o m p re ss io n of the surrounding normal hepatic tissue. The minor lesions of degeneration, hepatitis, ductule cell proliferation, necrosis and focal lymphoid infiltration seen in the control a n im a ls and the T-I and T - II groups a r e lesions of spontaneous disease and a r e not related to the A ro c lo r 1254. This level of liver disease is not unusual in rats of this age. Hyperplasia of the transitional epithelium was p re s e n t in the u r in a r y bladder of animals from the co n tro l group and each of the test groups. This lesion was usually associated with cystitis. The m o st . severe changes were associated with the p re se n c e of m ineralized calculi in the bladder lumen. All of the other lesions in o th e r tissu es found in these animals are related to spontaneous disease and they are not unusual for old rats. Donovan E . G ord o n , D. V. M. . P h . C. EHp l m a t e . A m e r i c a n C olle ge o f Veterinary Pathologists ACM CC3011 000 414 415 * r' Number of Animals 11 M a l e s > V. 5s o* oo Oi>'i TABLE xliii [TEST M ATERIA L: A r o c l o r 1254 ii Tw o-Y ear Chronic Oral Toxicity Study - Albino Rata Histopathologic Changea .Final Sacrifice Organ Examined Group: Control , Findings i Incidence Heart Trachea Lung Liver i Stomach Spleen, Kidney Testes, 1l Prostate Adrenal Gland 11i Brain |. Ev' e i Chronic focal myocarditis Tracheitis Chronic respiratory disease Focal degeneration puctilc hyperplasia Chronic gastritis Hemosiderosis { Chronic nephritis Testicular degeneration Periarteritis . Inflammation -Telangiectasis ,J Nodular hyperplasia * Focal fatly change ! 'Focal encephalitis Inflammation * | 3 3 9 1* 2' 1. 3 a* 2 2 1 1 1 1 1 2 t II tP O nt m Average Grade 1.0 1.0 1.0 1.0 1.0 1.0 1.0 1.0 1.0 1.0 1.0 1.0 1.0 1.0 1.0 1.0 a -u 000 Number of Animals 14 F e m a l e s >o X I TABLE'XLIII continued T E ST MATERIAL: A ro c lo r 1254 T w o-Y ear Chronic Oral Toxicity Study - Albino Rate Histopathologic Changes Final Sacrifice Group: Control Organ Examined Findings Heart Trachea lung . Liver Stomach Colon Spleen Uterus Kidney Ovary % Chronic focal myocarditis Tracheitis Chronic respiratory disease Focal lymphoid infiltration Ductile hyperplasia Focal hepatitis Non-lipid vacuolar change Chronic gastritis Acanthosis - squamous epithelium Parasites Hemosiderosis Metritis Chronic nephritis Focal lymphoid infiltration Cyst . Incidence 7 3 13 3 2 1 2 3 5 1 4 3 6 1 1 n u Average Grade 2.0 1.0 1.0 1.0 1.0 1.0 0. 5 *1.0 1.0 1.0 1.0 2.0 1.0 1.0 1.0 f -I c m 000 P i t O rsniirr TADLCtXUtl continued T E S T ,MA TE [UAL: A r o c l o r 1254 , Ttyo-Year .Chronic O ral Toxicity Study - Albino R ats Histopathologic Changes Number of Animals 14 F e m a l e s Organ Examined Adrenal Gland Salivary Gland Final Sacrifice Group: Control4 Findings Telangiectasis Nodular hyperplasia pltcocliromocy toma Chronic fibrosis and atrophy Incidence 9 4 1 1 All other tissues and organs w ere norm al histologically. 0.5 s'mlnimal 1.0 s slight Grading System 2. 0 = m ild 3.0 = moderate 4.0 = severe 5.0 = extreme AD* 00361* ` Average Grade 1.0 1.0 1.0 1.0 It l" K i Number of Anirr'-ls 11 O ?, o* oo oo UOJ' e l ' ` T A B L E XLIV* T E S T M ATERIAL: A ro c lo r 1254 'Tw o-Y ear Chronic Oral Toxicity Study - Albino Rata Histopathologic Changes Final Sacrifice Group: I ppm Organ Examined Heart Trachea Lung Liver Pancreas Stomach Spleen Kidney Testes, Findings Chronic focal myocarditis Tracheitis Chronic respiratory disease Congestion Ductile hyperplasia Vacuolar change (fatty) Necrosis Chronic focal pancreatitis Periarteritis Chronic gastritis Acanthosis lllcer Hemosiderosis Chronic nephritis Testicular degeneration Average _______ I n c i d e n c e _______________Gra<lc 5 1.0 6 1.0 9 1.0 1 1.0 1 0.5 1 1.0 1 3.0 2 1.0 1 2.0 2 1.0 2 1.0 1 l. 0 5 2. 0 a 2. 0 2 1.0 '* r Number of Animals 11 Males T A B L E X'LIV co ntinued T E S T M A T ER IA L : A ro c lo r 1254 Tw o-Year Chronic Oral Toxicity Study - Albino Rata Histopathologic Changes Final Sacrifice Group: I ppm * Organ Examined Testes (continued) Prostate Seminal Vesicle Pituitary Adrenal Gland Parathyroid Gland Findings Periarteritis Focal calcification inflammation Inflammation Congestion Nodular hyperplasia Hyperplasia Incidence 2 2 6 2 1 1 1 22 F e m a le s Heart Trachea Lung Liver 40* *< u Chronic focal myocarditis Tracheitis Chronic respiratory disease Focal lyniphoidinfilhration 12 16 17 3 tu Average Grade 1.0 2.0 2.0 2. 0 1.0 1.0 1.0 1.0 1.0 2.0 1.0 L-.II 000 419 A TABLE XLIV continued T E ST M A TER IA L: A ro c lo r 1254 Tw o-Y ear Chronic O ral Toxicity Study - Albino Rata Histopathologic Changes Final Sacrifice Group; 1 ppm ' Number of Aninv Organ Examined Findings ADM 0 03 b 17 22 Fri.:, i * O Liver (continued) Stomach Colon Spleen Lymph Node Kidney Uterus Pituitary Adrenal Gland Ductile hyperplasia Vacuolar change (fatty) Congestion Necrosis Focal inflammation and acanthoala Parasites Hemosiderosis Abscesses Chronic nephritis Focal calcification Metritis Hyperplasia Telangiectasis Nodular hyperplasia Congestion t Focal fatty change 4Kk) O .11 o t h e r t i s s u e s and o r g a n s w e r e n o r m a l his to lo g ic al ly . Grading System Incidence 2 1 1 1 4 2 a 1 8 3 6 4 17 1 1 3 & E O Average Grade 1.0 2.0 1.0 1.0 2.0 1.0* 2.0 * 1.0 1.0 1.0 1.0 1.0 1.0 1.0 1.0 1.0 -I 4- z' Number of Animals 12 M ale s > o * o o l*J ao-- GO TADLE XLV T E S T M A TEIU A L: A ro c lo r 1254 ' T w o-Y ear C hronic O ral Toxicily Study - Albino R ata Histopathologic Changes Final Sacrifice G ro u p : 10 ppm * Organ Examined H eart' Trachea Lung Liver 1 i . Pancreas Stomach Spleen 1Kidney T ables Findings Chronic focal m yocarditis Tracheitis Chronic respiratory disease Pneumonia V acuolar change (fatty) Ductile hyperplasia Hypertrophy Focal degeneration Chronic focal pancreatitis Chronic gastritis and acanthosis H em osiderosis Chronic nephritis Testicular degeneration Incidence 7 9 II 1 3 i i i i l 2 7 1 A verage G rade 1.0 2.0 1.0 1.0 1.0 1.0 .0 .5 0.5 3.0 1.0 1.0 I. 0 2. 0 m 000 ACM 003* 000 422 f Number oi Animals 12 Males. T A B L E -XLV co n tin u e d TEST MATERIAL: A roclor |254 1T w o - Y e a r C h r o n ic O r a l T o x ic ity Study - A lb in o R a t a Histopathologic Changes Final Sacrifice G ro u p : 10 p p m ' Organ Examined T estes (continued) P ro sta te Seminal Vesicle A drenal Gland. Thyroid Gland Findings P e ria rte ritis Inflam m ation Inflam m ation Telangiectasis Focal fatty change Focal hyperplasia Incidence 1 2 2 2 1 1 12 F e m a l e s . H eart Trachea Lung Liver i i Chronic focal m yocarditis T rach eitis Chronic respiratory disease Focal lymphoid Infiltration V acuolar change (fatty) Nodular hyperplasia 3 7 12 4 1 1 v A verage G rade 3.0 2.0 1.0 1.0 1.0 1.0 1.0 1.0 J. 0 1.0 1.0 3.0 a O I * TABLE-XLV continued T E ST M ATERIAL: A ro c lo r 1254 T w o-Y ear Chronic O ral Toxicity Study - Albino Rata Histopathologic Changes Final Sacrifice G ro u p : 10 p p m N um ber of Animals Organ Examined Findings Incidence A OP. C C 3 6 2 0 12 F e m a le s O Liver (continued) Colon Spleen Lymph Node Kidney U terus Pituitary Adrenal Gland Parathyroid Gland Thymus Ductile hyperplasia Telangiectasis llcpatom a Parasites H em osiderosis Congestion Chronic nephritis Focal calcification M etritis Adenoma Hyperplasia Telangiectasis Hyperplasia Abscess T 4 A ll other, t i s s u e s and o r g a n s w e r e n o r m a l h is to lo g ic a l ly . K> Grading System 2 1 1 1 2 1 3 2 1 3 1 9 1 1 A verage G rade 1.0 1.0 1.0 1.0 1.0 1.0 .2.0 1.0 1.0 1.0 1.0 1.0 1.0 1.0 N um ber of A nim als 11 Male* TABLE XLVI T E ST M ATERIAL: A ro c lo r 1254 Tw o-Y ear Chronic O ral Toxicity Study - Albino Rata Histopathologic Changes Final Sacrifice G ro u p : 100 ppm* Organ Examined Heart Trachea Lung Liver Small Intestine Spleen Lymph Node Findings Chronic focal m yocarditis Tracheitis Chronic respiratory disease Edema Focal degeneration Vacuolar change (fatty) Ductile hyperplasia Hypertrophy Focal lymphoid infiltration Portal fibrosis Focal inflammation H em osiderosis Congestion Incidence 3 4 10 3 2 3 3 1 1 1 1 2 1 A verage G rade 2.0 1.0 1.0 2.0 1.0 i;o 1.0 2.0 1.0 2.0 1.0 1.0 1.0 AOM 0 0 3 6 2 1 Number of A nim ala 11 Males TADLE XLVI continued T E S T M A TER IA L: A ro c lo r 1254 T w o-Y ear Chronic O ral Toxicity Study - Albino R ate Histopathologic Changes Final Sacrifice G ro u p ; 100 p p m Organ Examined Kidney * Testes P ro sta te Adrenal Gland. Thyroid Gland Findings Chronic nephritis P e ria rte ritis Inflam m ation Telangiectasis Congestion H y p erplasia Incidence 8 1 5 2 i 1 A verage __________ G r a d e 1.0 1.0 2. 0 1.0 1.0 1.0 1$ F e m a l e s O O H eart Trachea Lung Liver 1 Chronic focal m yocarditis T rac h e itis Edema Focal hypertrophy H ypertrophy F o c a l h y p e r p l%a s ia * 5 6 13 1 1 5 1 1.0 1.0 2.0 2. 0 1.0 3.0 1.0 AM 0036 r T A B L Et X L V I co n tin u e d T E ST M ATERIAL: A ro c lo r 1254 Tw o-Y ear Chronic O ral Toxicity Study - Albino Rata Histopathologic Changes Final Sacrifice G ro u p : 100 pprrv Number of Animal* IS Female* Organ Examined Liver (continued) S tom ach Spleen Kidney 9 ACM 0 C 3 6 2 3 Uterus Pituitary Pituit Adrenal Gland O Findings Ductile hyperplasia Vacuolar change (fatty) Nodular hyperplasia Hepatoma Chronic gastritis Acanthosis Inflam m ation H em osiderosis Chronic nephritis Focal calcification M etritis H y p erplasia Nodular hyperplasia Telangiectasis Nodular hyperplasia Congestion Incidence 2 10 1 1 2 4 1 5 5 1 3 4 I 3 1 1 M All other tissu e s and organs w e re n o rm al histologically. Ch G ra d in g S y s te m A verage G rade 1.0 1.0 1.0 1.0 1.0 1.0 1.0 1.0 1.0 1.0 1.0 1.0 1.0 1.0 1.0 1.0 no Of 42 7 TABLE XLVII TEST M ATERIAL: A ro c lo r 1254 Tw o-Y ear Chronic O ral Toxicity Study - Albino Rate lliatopathologlc Chance of the U rinary B ladder Final Sacrifice Group C T -1 (1 ppm ) Number Sex Examined M 11 F 14 M 10 F 19 Findings* P ro tein ac eo u s plug (34) Focal epithelial hyperplasia (62) P ro tein aceo u s plug (326) Diffuse epithelial hyperplasia associated with diffuse cy stitis (377) T-Il (10 ppm) M 13 F 13 Diffuse epithelial hyperplasia and papilloma asso ciated with focal cy stitis (417) Diffuse epithelial hyperplasia associated. with diffuse cy stitis (419) None T-III (100 ppm) M 10 F 17 Focal epithelial h y perplasia a sso c iate d wll)i focal cy stitis (497) Focal epithelial hyperplasia (5) None l * Individual anim al number a a re shown in.parenthesis. Nj G ra d in e S y s te m 4* LO =m i n i m a l o r t r a c e 4.0 = severe Incidence 1 1 1 1 1 1 1 1 A verago G rade 1 4 %.. 3 4 2 2 1 - oo Z I O ' : 5 I tyLsbudi. G. D etailed D escription of Tum or Findings . Tum or data for individual anim als including location, weight a n d p a th o lo g ic c la s s if ic a t io n a r e p r e s e n t e d in T a b le XLV1II th ro u g h LX. J None of these tum ors could be re la te d to the ingestion of A ro c lo r 1254 and a re considered norm al for a random papulation of rats this age. AOM nniA (H 428 fr r Q6 Animal Number and Sex l-M 2-M 6-F -n-F 8-F 50-F 52-F 0 5 9 -F o o MiO - F o* 1 oU> fOvj' -F ` TADl^E XLVIII TEST M ATERIA L: Aroc lo r 1254 T w o-Y ear Chronic O ral Toxicity Study - Albino Rata D escription of Tum or Findings t Location Abdomen Pancreas Abdomen W eight (gram a) - 11.76 64.98 35.09 Group: Control ________________________________________________________ :_______________________________________________ M onths Size on (cm) C lassificatio n Tost - M am m ary fibroadenoma 24 - Adenocarcinoma 24 8x1x2 5x2x2 M am m ary fibroadenoma M am m ary fibroadenoma 24 t ! Lymph node Abdomen Abdomen Abdomen Abdomen *drenal gland > - m 2x2x1 2x2x3 - Reticulum celt sarcom a M am m ary fibroadenoma M am m ary fibroadenoma M am m ary fibroadenoma M am m ary fibroadenoma Phcochrom ocytom a 24 24 7 24 24 Abdomen Abdomen 88.92 230.00 13x11x4 13. x 11 x 4 Fibroadenom a 1 M ammarv adenoma 24 1A 0 0 0 429 r Animal `j u m h e r *-.nd Sox . 11 - F V 12-F 1.3- F * * ! ocation Abdomen Ahdonr en Abdomen TABLE XLIX continued TE ST M ATERIAL: A roclor 1254 T w o-Y ear Chronic O ral Toxicity Study - Albino Rata D escription of Tum or Findings Group; I ppm W eight (gram s) - - -. Size (cm) - - - C lassificatio n y M ammary adenocarcinoma / {, M a m m a r y f ib r o a d e n o m a Lipoma A J*) */ <*** ^ iW* ** *^4. h. *i c C2*J cr u> o i * n M onths on Teat 4 24 24 Animal Number and Srx l-M 2-M 3-F 4-F 5-F 6-F 7-F : 8-F 9-F o> * o TAQLE XLIX TEST M A TER IA U A ro clo r 1254 Tw o-Y ear Chronic O ral Toxicity Study - Albino Rate Description of Tum or Findings Group; 1 ppm Location Weight (grama) Parathyroid gland - Pituitary - Abdomen - Abdomen Abdomen Parathyroid gland % 8.97 - Abdomen 8.86 Abdomen Doraal cervical region 65.0 209 Pituitary Abdomen a Abdomen - Size _______<<=">) 2x2x2 3x2x2 13 x B x 2 i* m1. C la ssific a tio n V Adenoma *' C h ro m o p h o b e a d e n o m a 1- M a m m a r y f ib r o a d e n o m a V " M am m ary fibroadenoma IX 1M ammkry adenom a-"'' V Adenoma u i M ammary fibroadenoma * M a m m a r y f ib r o a d e n o m a i ' ' ^Fibroadenoma ; `i C h ro m o p h o b e a d e n o m a w M ammary fibroadenoma ^ *' M a m m a r y fib r o a d e n o m a '" M onths on Test 24 23 24 24 24 24 24 24 ya o & TABLE L I TEST MATERIAL] A ro clo r 1254 T w o-Y ear Chronic O ral Toxicity Study - Albino Rata Description of Tum or Findings G roup: 10 p pm \nim al Cumber nd Sex 1-M Location Pancreaa W eight (grama) - Size (cm) C la ssific a tio n y I^Adenoma 1 2-M Parathyroid gland - 3-F Abdomen - - 4-F Thyroid Abdomen 5-F Abdomen -_ - -- '"Adenoma U- "Mammary fibroadenoma ^ 1 Adenoma /"Mammary fibroadenoma ^ M am m ary fibroadenoma 6-F Abdomen Abdomen 86 29 7x8x3 5x5x2 i s F i b r o a d e n o m a 4-" Fibroadenoma * 7-F Pituitary - - VChromophobe adenoma 8-F oDC 9-F o o o-r sO Abdomen Pituitary Adrenal gland 28 m 7.8 6x5x3 * - /Fibroadenom a l^* (/Chromophobe adenoma t / T e l a n g l o m a / t t,'1 O M onths on Test 24 24 24 24 24 24 24 24 22 g 24 W ijw rr; 1 53 a I a Animal Number and Sex 1-F 2-F 3-F 4-M 5-M 6-F 7-M a-F O O 9 *F o,, 10-F * X*. oo tUoJ utOu>' Location Abdomen Abdomen Abdomen Pancrcaa Abdomen Abdomen Lymph node Pancrcaa Lymph node ` Abdomen D TABLE LI TEST MATERIAL! A ro clo r 1254 T w o-Y ear Chronic O ral Toxicity Study - Albino Rata Description of Tum or Findings G roup; 100 ppm W eight (trin ili 48.12 % - 13.0 52.0 Size (cm| 5x4x3 18 x 4 x 3 *1x2x1 3x3x3 a x 5X2 C lassificatio n [/Fibrom a J/' I f M amm ary fibroadenoma ^ is M am m ary fibroadenom a 1 / [/Adenoma /' F i b r o a d e n o m i ' j M am m ary fibroadenoma L Reticulum cell sarcom a / [/'A denocarcinom a j-- [/Lymphosarcoma K V F ib r o a d e n o m a I f 0" % M onths on Test 24 24 24 24 24 24 13 24 24 24 OO -4 434 Animal Number and Sex 11 - F 12-F 13-F 14-F 15 - F Location Abdumcn Abdomen Abdomen Abdomen Abdomen TABLE L! continued T E ST MATERIAL: A ro c lo r 1254 Two*Year Chronic O ral Toxicity Study - Albino Rata Description of Tum or Flndlnga G roup: 100 ppm W eight (gram s) 141.9 42 209.0 14. 6) Size (cm) 8x8x 2 6x 4x 2 10 x 9 x 3 - C laaslflcatlon Is' Mamma ry flbroadcnomat^" M a m m a r y fib r o a d e n o m a i-- is Fibroadenom a / M ammary fibroadenoma l Fibroadenoma O O o X I ooL*J boi ir i i c < < M onlha on Test 24 24 24 24 24 Ooo0 j-d+i rtfLud. B I u - I E S T 2a*vuza>u*i, 2*c. R E P O R T TO MONSANTO COMPANYTWO-YEAR CHRONIC ORAL TOXICITY WITH AROCLOR 1242 IN ALBINO RATS NOVEM BER 12, 1971 " IBT NO. B7298 I. Introduction At the req u est of Monsanto Company, a tw o-year chronic o'ral toxicity study was conducted using albino ra ts to determ ine the potential toxicity of A ro c lo r 1242, Lot No. AK-255. The following re p o rt p r e s e n ts the results of this investigation. ( CC3b3 OOC 435 B I C T E S T JzacyuiLyM* ^ IL Sum m ary A tw o -y e a r chronic toxicity study w u conducted using albino rats fe d d ie ts c o n ta in in g 1 (T -I), 10 (T -II) o r 100 (T -III) ppm A ro c lo r 1242. Food consumption and body weight gains were not altered at any le v e l of A ro c lo r 1242 fed. The n u m b e r of a n im a ls dying and the tim e at which the deaths o ccu rred did not vary among th test and control g ro u ts. H em atological and clinical blood chem istry studies conducted after 3 ( 6, 9 , 12, 18 o r 24 m o n th s did n o t r e v e a l any a ffe c ts re la te d to A ro c lo r 1242. U rine analyses failed to re v e a l any differences betw een test and control animals. A t s a c r i f i c e a f t e r 3, 6 o r 12 m o n th s on t e s t , o r g a n w eights, o r g a n to body weight and organ to b rain weight ratio s disclosed several random ly o c cu rrin g intergroup d ifferences. The lack of any consistent dose re la te d resp o n se and the absence of any d eleterio u s histopathologic change confirm that these differences w ere not re la te d to the ingestion of A ro c lo r 1242. At the final sac rifice after 24 months on te st, the liver weights and liv e r to body weight o r b rain weight ratios w ere significantly elevated in fe m a le s fro m the T-III group. H istologic exam ination of the liv e rs from the T-III roup revealed several anim als with vacuolar change. This lesion is m orphologically indicative of fatty degeneration. Specific fat stains confirm ed the presence of fat in these vacuoles. Focal h y p e rtro p h y and fo cal h y p e rp la s ia w e re a lso found in the liv e rs fro m a n im a ls fed A ro c lo r 1242. ADM 0 0 3 6 3 3 0 0 ' 436 B I ' T E S T za&akuM. 2 H y p erp lasia of the u rin a ry bladder was found in anim als from the c o n tro l group and from each of the te st groups. This hyperplasia w as u sually asso ciated w ith cystitis. T he in cid en ces and types of all tum ors w ere about the sam e in all g ro u p s, including the control group, and are considered norm al for rats of this age. Respectfully subm itted, INDUSTRIAL BIO -TEST LABORATORIES, INC. R eport prepared by: P h ilip s \ S m ith , B . S. A ssistant Toxicologist Rat Toxicity R eport approved by: 4/ r ji 7 * -- P a u l L. b r i g h t , P h. D. Section Head, Toxicology M. L , Keplinffer, M anager, Toxicology N ovem ber 12, 1971 lm :bb:hp Jcjl ADM 0 0 3 6 3 * 001 437 uaf B I ~ - T E S T 2zo*eJa*+A. J, 4 HI. P ro ced u re A. E xperim ental Animals The anim als employed in the te st w ere C harles River strain^ albino r a t s . F o u r hundred r a ts (200 m a le and 200 fem ale) w e re 's e le c te d L for the experim ent, ear-punched with the anim al number assigned and housed individually in standard wire-bottom ed steel rat cages. Each cage b o re a c o lo r-c o d e d c a rd identifying the r a t with re s p e c t to p ro je c t num ber, dietary level assignm ent, individual anim al number and sex. B. Organization of Groups # A s tr u c tu r a l outline of the e x p erim en t is given in Table I. TABLE I TEST MATERIAL: A ro c lo r 1242 T w o-Y ear Chronic O ral Toxicity Study - Albino Rats (. Outline of Experim ent* TOUO Control T-I T-U T-HI N um ber of A nim als Male Fem ale 50 50 50 50 50 50 50 50 dietary Level ftrom) None A dm inistered 1 10 100 * C harles R iver Breeding L aboratories, North Wilmington, M ass. 0033 0 0 1 438 S I O S T Jaoo+rJo+i+i, $*c. 5 C. D iets, Feeding and Food Consumption A ll d ie ts w e re p r e p a r e d in the c e n tra l d ie t ro o m of this la b o ra to ry . The b asic ration from which all diets w ere constructed w as a stan d a rd , pulverized stock ra t ration*. The diet for any given L t e s t group w as p re p a re d by blending the calculated am ount of A roclor 1242 with a p re -w e ig h ed portion of the stock ra tio n in a Hobart m ix er. F re s h diets w ere prepared each week and every rat was offered a n am ount of food sufficient for ad libitum feeding. Food consum ption was recorded for five ra ts of each sex in every group w eekly for the f ir s t th ree months and m onthly for the next nine m onths. T hereafter, periodic spot checks w ere made. D. Bodv Weights and Weight Cains i Initially, the body weight of each ra t in every group was d e te r m ined and recorded. Thereafter, individual weighings were made weekly f o r the f i r s t 13 w e e k s and m o n th ly th e r e a f te r u n til the conclusion of the investigation. E. M ortality and Reactions Checks for m ortality and abnormal behavioral reactions were m ad e daily throughout the investigation. F* H e m a to lo g ic Studies and U rine A n aly ses Blood studies, including determinations of hemoglobin concentra tion, h e m ato crit value, erythrocyte count and both total and differential e * Purina Rat Chow, Ralston Purina Company, St. Louis, M issouri. ACK C0363 00( 439 leukocyte count, w ere conducted upon five m ale and five fem ales from b o t h the c o n tro l and 100 ppm g ro u p s. I U rin e a n aly ses for the presence of g lu c o se , albumin, m ic r o s c o p ic e le m e n ts , and d e term in a tio n s of pH and s p e c ific gravity w ere con ducted upon u rin e sam ples at the same time in terv als and from the sam e a n im a ls as those em ployed for the blood studies. T h e s e s tu d ie s w e r e conducted a f te r 3, 6, 9, 12, 13, and 24 m onth o f feeding. G. C linical Blood C hem istry Studies D e term in atio n s of blood u rea nitrogen concentration (BUN), s e r u m alkaline phosphatase activity (5AF), fasted blood glucose concen tra tio n and se ru m glutam ic-pyruvic transam inase activity (SGPT) w ere f c o n d u c te d a f t e r 3, 6, 9, 12, IS , and 24 m o n th s o f fe e d in g upon the s a m e r a t s as w ere the routine blood studies. H. Pathologic Studies When an anim al succumbed during the te st period, a gross autopsy w a s p erfo rm ed . In those cases when postm ortem autolysis was not ad- . anced, re p re se n ta tiv e tissu es w ere taken and p re s e rv e d in ten percent form alin solution for possible future histopathologic study. A f t e r 3, 6 , a n d 12 m o n th s of te s t in g , fiv e a n i m a l s of e a c h s e x fro m each group w ere sacrificed and subjected to complete gross pathologic exam inations. ACM 001 C03637 iii 440 - a L B I l T i 5 T Jaaaxalauad. % 7 A bso lu te org an w eights w e re re c o rd e d and organ to body w eight and organ to b ra in weight ratios computed. The following organs wereincluded: liv er, kidneys, spleen, gonads, h eart and brain. T hese'data w e re then subjected to s ta tistic a l an aly sis. An A nalysis of V ariance was conducted first and significant effects disclosed by that treatm ent were fu rth e r studied by "t" -te s ts . Com plete m icroscopic examinations w ere conducted upon the t i s s u e s a n d o r g a n s f r o m a ll c o n tro l and 100 p p m g ro u p r a t s s a c r i f i c e d a f t e r 3, 6, o r 12 m o n th s of te stin g . The follow ing tis s u e s and o r g a n s w e re included in the exam inations: h eart, trachea, lungs, liver, p a n crea s, esophagus, stom ach, sm all intestine (duodenum, jejunum and ileum ), caecum , colon, spleen, lymph nodes, kidneys, urinary bladder, gonads, p ro state gland, sem inal vesicles, uterus, pituitary gland, adrenal glands, saliv ary glands, thyroid gland, parathyroid glands, skeletal m uscle, sternum , bone, peripheral nerve, spinal cord and brain (cerebrum , cerebellum and pons). The aforem entioned tissues were p rep ared and stained with H em atoxylin-Eosin stain. A fter two y ears of feeding, all surviving anim als w ere sacrificed and subjected to complete gross pathologic examination. The absolute weights of the liver, kidneys, spleen, gonads, h eart and brain w ere recorded immediately after sacrifice. 1 AO* B I C T E S T a&ui/ivut, 2* S M icrosco p ie exam inations w ere conducted upon tissues and o rg an s fro m selected anim als dying during the ex p erim en t and upon all an im als from the final sacrifice. T issues and organs examined w ere the sam e as those previously m entioned. Z. T u m o r In c id e n ce and C la s s ific a tio n A tabulation, of the incidence of tu m o r fo rm atio n in each group w as m ade a t the conclusion of the investigation, tn addition, tum or data for individual anim als including location, weight, size and pathologic classification were recorded. t t ACm c <3639 0 0 ' 442 H fa U '- d B I 0 - T HS T Js U d * * * . f a 9 IV. R esults A, Body Weights m d Weight Gains The m ile and fem ile body weight d i t i i r e issem b led in T ib le I T h e d i t i in th is t i b l e i l s o in c lu d e the 3, 12 and 24 m o n th t o t i l w e ig h t g i i n s . T h e re w e re no sig n ific in t effects a ttrib u tab le to the ingestion of the compound. i / AD* orr** 00' 443 I T A B L E II T E ST M ATERIA L: A ro c lo r 1242 T w o-Y ear Chronic O ral Toxicity Study - Albino R ats Body Weight and Total Weight Gain Data Sum m ary of Mean Values dietary Level of A r o c l o r 1242 (ppm) Sex 0 12 Body Weight (grams) Week: 3456 7 85 10 I I 12 13 lontrol * I 10 M 133 203 245 313 362 373 391 409 424 440 440 449 467 474 F 136 178 190 219 244 254 259 265 270 276 281 278 290 288 M . 133 205 254 329 367 377 397 417 432 451 462 4SI 493 508 F 136 173 197 226 240 253 268 292 278 280 284 284 288 292 M 133 202 251 332 362 372 392 414 428 454 475 481 500 509 F 136 176 196 226 244 254 268 288 276 280 281 284 260 285 100 M 133 203 248 320 365 376 398 416 427 439 453 467 478 487 F 136 173 194 225 251 264 272 290 274 275 279 285 283 286 90-Day Total Weight G a in (g r a m a / r a t ) 341 152 375 156 376 149 354 150 %t ir < i I O o. > x* o l.Uu 1 o o _ c TABLE It continued T E S T M A T E R IA L : A r o c lo r 1242 T w o-Y ear Chronic O ral Toxicity Study - Albino Rats Body Weight and Total Weight Gain Data Sum m ary of Mean Values lietary Level of A ro c lo r 1242 (ppm) Sex 4 5 6 Body Weight (grama) Month: 78 9 10 1 1 12 Control M 498 533 565 593 622 642 667 688 701 F 297 310 322 333 345 359 374 383 390 1 M 525 555 580 614 640 665 690 707 743 F 302 321 326 333 342 355 367 381 388 10 M 525 553 579 6 0 0 627 641 674 693 706 F 299 312 328 333 346 354 365 375 384 100 M 504 533 551 580 . 608 633 662 686 711 F 290 307 321 333 347 360 374 388 379 O o JtCn* >o o o LM 12-M onth Total Weight Gain (gram a/rat) 568 254 610 252 573 248 578 243 U . t 4 46 Hetary Level of A ro c lo r 1242 (ppm) Sex Control M F 1M F 10 M F 100 M F T A B L E II co n tin u ed *- T E ST M ATERIA L: A ro c lo r 1242 T w o-Y ear Chronic O ral Toxicity Study - Albino Rata 03 O Body Weight and Total Weight Gain Data Sum m ary of Mean Values m i/i Body Weight 24-M onth (gram a) T otal W eight Month: G ain 13 14 15 16 17 IB 19 20 21 22 23 24 ( s r a m a / r a t ) r 728 746 759 768 769 767 760 758 739 726 687 642 509 426 444 466 481 500 515 533 547 551 550 564 551 415 767 785 805 800 804 810 800 804 802 .800 808 759 428 443 479 468 500 514 523 531 535 538 535 532 626 396 740 750 r 763 780 784 780 785 783 780 774 796 734 393 406 422 441 462 480 500 519 529 533 537 547 6o 1 4M 724 728 723 735 728 736 733 730 725 715 743 652 428 447 463 467 472 484 481 484 485 490 487 4 8 s ' 519 349 B I 0 T HS T JiJ\aU*u* jke. 13 B, Food Consumption The data obtained fro m the food consumption m easurem ents c o n d u c te d d u r in g th e f i r s t 12 m o n th s of feeding a r e s u m m a r iz e d in T a b le III. - T hese data revealed no significant difference between test L and control ra ts. The periodic checks conducted during the rem ainder of the investigation also rev ealed no outstanding differences betw een te s t and control anim als. ACP 00 447 r 7 l S21 0 Ia T A B L E III T E S T M ATERIA L: A ro c lo r 1242 I! T w o-Y ear Chronic O ral Toxicity Study - Albino Rats Food Consumption Data Sum m ary of Mean Values Food Consumption letary Level of (gram s/rat/sev en days) Average Food A ro c lo r 1242 Week: C onsum ption (ppm Sex 1 2 3 4 5 6 7 8 9 10 11 12 13 ( g r a m s / r a t / s e v e n davi Control M 94 1 6 0 175 163 137 137 225 197 204 188 191 194 196 F 73 140 144 107 104 133 208 154 165 151 146 150 151 174 140 1 M 129 154 165 201 120 191 206 210 232 185 188 189 190 F 91 109 120 164 110 166 124 137 172 140 145 148 145 167 136 10 M 113 132 142 201 120 193 183 184 184 192 196 191 193 F 96 114 147 131 1 10 149 102 124 166 144 143 138 144 171 131 100 M 117 149 185 230 161 172 199 172 227 192 186 190 190 F 89 118 129 189 98 114 142 135 180 145 142 139 140 182 135 O On 00 M a l B I 0 - T E S T M AD- 00( 449. etary Level of A roclor 1242 ___ (ppTM> Sex 4 Control M 196 F 148 1 M 180 F 152 10 M 184 F 139 100 M 179 F 163 TA B LE III continued T E S T M ATERIAL: A ro c lo r 1242 T w o-Y ear Chronic O ral Toxicity Study - Albino Rata Food Consumption Data Sum m ary of Mean Values Food Consumption (gram a/rat/seven days) Month: 5 6 7 8 9 10 11 12 189 168 147 163 184 179 170 187 154 136 121 138 152 160 141 127 181 170 189 190 189 196 192 208 159 141 154 150 148 143 145 159 164 168 197 193 1RS 191 186 177 147 136 151 144 145 138 142 130 192 189 200 196 186 184 187 167 141 160 148 145 144 147 146 145 Average Food C onsum ption (gram s/rat/seven days) 176 142 188 150 183 14! 187 149 Ln *"--fc** ''* 3 1 0 - . S T la C. M o rta lity and R eactions A freq u e n cy d istrib u tio n of n atu ral deaths o c c u rrin g during the in v e stig a tio n a p p e a rs in T able IV. No u n to w a rd b e h a v io ra l re a c tio n s w ere noted am ong any of the a n im a ls em ployed in the investigation. aCH 0 0 3 0 * 7 O O t 450 ! >( vm tTfovm j- i ; 3 ) * O ! 9 Dietary Level <ppm > Sex Control M F 1M F 10 M F 100 M F T A B L E IV T E ST M A TER IA L: A ro c lo r 1242 T w o-Y ear Chronic O ral Toxicity Study - Albino Rata M ortality Data Frequency D istribution of Deaths 1-3 4-6 Frequency Distribution of Deaths M onths: 7-9 10-12 13-15 16-18 19-21 000 2 17 6 000 1 3 5 7 20 1 1 3 3 7 10 0 1 3 2 4 000 2 17 5 0 11 2 0 2 5 000 3 3 6 7 0 I0 1 2 4 3 22-24 9 3 8 5 8 5 10 9 Total Dead N u m b e r T e s t e d 1* 25/35 19/35 25/35 16/35 23/35 16/35 29/35 25/35 * Does not include the five anim als of each sex from every group sacrificed for interim pathologic a tudie s . ad* OOl 451 Y BIO TES T 18 D. H em atologic and C linical Blood C hem istry Studies M ean m ale and fem ale hem atologic and clinical blood chem istry i d a t a a r e s u m m a r i z e d In T a b l e s V t h r o u g h DC. V alues for all p a ra m ete rs investigated w ere within the norm al range for the albino rat. No significant differences between test and control values w ere observed. Q O i b 1*9 0 0 452 ssuluib-isd B I 0 - T E 5 T Jaa. 0 0 453 C D ietary Level (ppm) Sex 3 Control- M 14*0 F 9. 1 TABLE V T E ST M ATERIAL: A ro clo r 1242 T w o -Y ear C hronic O ral Toxicity Study - Albino R ata H em atologic Data Sum m ary of Mean Values Total Leukocyte Count (thousands/m m ^) M onth: 6 9 12 18 15.4 10. 1 11. 1 9 .9 9.4 6.7 7.6 8.8 24 11.2 7.6 Erythnocyte Count (m illions/m m ^) M onth: 36 9 12 18 8. 1 8 .2 8 .7 8 .8 7.6 7.5 7.7 7.3 7.8 7.7 24 6.5 6 .8 M IS.6 10.4 9.7 * 12.5 9.4 9.3 100 F IS.6 9.0 . 7.7 7 .8 6. 2 6. 4 7.7 8.0 8.0 7.6 7.4 7.6 7.6 7.1 7. 9 7 .0 7.7 7.2 o D vO > * n * r D ietary Level (ppm) _ Control Sex M F 3 17. 1 16.9 T A B L E VI T E S T M ATERIA L: A ro clo r 1242 T w o-Y ear Chronic O ral Toxicity Study - Albino Rats H em atologic Data Sum m ary of Mean Values Hemoglobin Concentration (g/100 ml) Month: 6 9 12 18 17.7 15.7 17.6 16. 1 17.4 14. 1 16.7 16.4 24 13.6 14.6 H em atocrit Value 36 (%) Month: 9 12 18 51 49 48 50 41 50 47 43 46 42 M 16.8 16.9 14.2 15.4 15.6 14.6 50 48 43 44 4.1 100 F 15.8 16. 1 13.6 15.7 15.5 14.8 46 48 41 41 40 L CP O m in r24 35 38 37 37 '-- > OO LI > * kJ r ro^j B I 0 T E 5 T a** 0 0 455 n D ietary Level <ppm > Control 100 T A B LE VII T E S T M ATERIA L: A ro c lo r 1242 T w o-Y ear C hronic O ral Toxicity Study - Albino Rata H em atologic Data Sex 3 M 82.0 F 83.0 6 86.6 86.6 Sum m ary of Mean Values D ifferential Leukocyte Count (Number of C ells p er Hundred) L ym phocytes Neutrophils Month: Month: 9 12 18 24 36 9 12 16 80.4 78.8 79.0 65.4 16.0 11.4 18.6 20.6 20.0 86.8 89.0 70.2 61.4 15.4 11.6 11.8 10.0 29.2 24 32.2 36.0 M 89.6 84.4 . 79.8 87.0 66.0 58.4 F 91.4 86.0 82.8 88.4 75.6 65.2 9.6 15.2 18.8 13.2 33.0 38. 8 8 . 4 I ) . 6 17.2 1 1. 2 2 3 . 0 3 3 . 6 *o> N o o Ui ,f^vt77e7, i S 3 l mO 18 D ietary Level tppm) Control 100 Sex 3 M 0.6 F 0.6 M 0.2 F 0.0 TADLE VII continued T E ST M ATERIA L: A ro c lo r 1242 T w o-Y ear Chronic O ral Toxicity Study - Albino Rats H em atologic Data Sum m ary of Mean Values Differential Leukocyte Count (Number of Cells per Hundred) M onocytes Eosinophils Month: Month: 69 12 18 24 3 6 9 12 0.0 0 .0 0.4 0 .2 1.2 1.4 2.0 1.0 0 .2 0.0 0 .0 0.0 0.2 1.0 1.0 1.8 1.4 . 0.2 18 . 24 0. 8 1.6 0 .6 1.6 0.0 0.0 6.4 0.0 0.8 0.0 0.0 0.2 0.0 0.2 0.6 0 .4 1.4 1.4 0. 2 0.4 0 .0 0 .2 1.0 2 .0 1.4 1.0 > o OX o o o U1 0\ u> n* _UL N I ia D ietary Level . .(ppm) C o n tro l Sex M F 100 M F TADLE VII continued T E S T M A T E R IA L : A r o c l o r 1242 T w o-Y ear C hronic O ral T oxicity Study - Albino Rata Hem atologic Data Sum m ary of Mean Values * D ifferential Leukocyte Count (Number of Cells p e r Hundred) Daaophils Month* 3 6 9 12 18 ________24 0.0 0.0 0.0 0.0 6.0 0.0 0.0 0. 0. 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 0.0 t 0.0 0.0 0.0 0.0 0.0 0.0 o o ft is i l AOM 0 0 3 6 5 fv BIO.TEST TABLE VIII T E S T M A TER IA L: A ro c lo r 1242 T w o-Y ear Chronic O ral Toxicity Study - Albino Rata C linical Blood C hem istry Data Sum m ary of Mean Values D ietary Level (ppm) Sex Serum Glutam ic-Pyruvic T ransam inase Activity > {Dade Units) M onth: 3 6 9 12 18 M 38 48 45 45 31 Control F 33 39 . 38 42 32 24 36 30 Blood Urea N itrogen Concentration (mgs %) Month: 3 6 9 12 18 17 15 12 10 14 14 16 15 9 13 24 15 II M 33 35 36 57 39 32 100 F 39 41 53 40 52 46 15 15 13 14 12 20 17 14 14 11 11 14 % LO J ) O' y i s 3 i * o i a t w t ." r f' D ietary Level (ppm) Sex 3 M 128 Control F 130 M 121 100 F 125 T A B L E IX T E S T M A TER IA L: A ro c lo r 1242 T w o-Y ear C hronic O ral Toxicity Study - Albino Rats C linical Blood C hem istry Data Sum m ary of Mean Values F asted Blood Glucose C o n cen tratio n (m g s %) - M onth: 6 9 12 18 123 89 88 138 118 115 92 134 24 99 109 Scrum Alkaline Phosphatase Activity (King-Armstrong Units) Month: 3 6 9 12 18 28.6 24.7 21.5 20. 3 16.2 18.9 17.8 10.6 11.9 16. 8 24 22. 3 20. 2 123 129 145 125 115 118 124 121 135 116 26.4 27.6 23.2 20. 3 17.4 9.6 25.8 11.7 16.7 11.6 13.2 14.2 uhi tNn VO BI0 TE5T ^c& 26 E. U rine Analyses Me&n m i l e end fem ale u rin e an aly ses data i r e su m m arized in T ables X through XIL No d ifferen c es w e re noted betw een the u rin e fro m control and te st in im ils i t the intervals of examination. aom On 001 460 BI 0 T s S T Jt ACH TADLE X T E ST M ATERIAL: A ro c lo r IZ42 Tw o-Y ear C hronic O ral Toxicity Study - Albino R ats Urine A nalyses Data Sum m ary of Mean Values k D ietary Glucose Level Month: (p p m ) _ S ex 3 6 9 12 18 24 Control M n n n n nn F n n n n nn M n n n n nn 100 F n n n n nn Albumin Month; 3 6 9 12 18 24 nn t. t + 2 43 n n n n t 42 n n I t 4 1 44 n n t t t 43 Key: n = negative t = t r a c e ; le e s than 30 m g / 100 m l u r in e 4 1 = 30 to 100 m g / 100 m l u r in e 4 2 = 100 lo 300 m g / 100 m l u r in e 4 3 = 300 Io 500 m g / 100 m l u r in e 44 = m o r e than 50 m g / 100 m l u r i n e B I O * ( E S T J.sJ* 462 TADLE XI T E ST M ATERIAL: A ro c lo r 1242 Tw o-Y ear C hronic O ral Toxicity Study - Albino Rata U rine A nalyses Data Sum m ary of Mean Values D ietary Level (ppm) .Control M icroscopic Elem ents Month: S ex 3 6 9 12 18 24 M N N N N N 4I F NNNNt t pH Month: 36 9 12 18 24 7 7 6. 7 7 6 8 8 7 8 66 M N N N N Nt 7 7 7 8 6 6- 100 F N N N N Nt 7 7 6 7 66 Key; N= N o r m a l t = trace; lesa than 5 p er low pow er field 4 1 = 5 to 10 p e r low p o w e r fie ld 4 2 = 10 to 15 p e r low p o w e r fie ld 3 = m o r e th a n 15 p e r low p o w e r fie ld U) <r N 00 BI 0 T E S T M D ietary Level <ppm > _ Control 100 T A D L E XII T E ST M A TERIA L: A ro c lo r 1242 T w o-Y ear C hronic O ral Toxicity Study - Albino Rats U rine A nalyses Data Sum m ary of Mean Values Specific Gravity Month: S e x 3 6 9 12 M 1.029 1.022 1.033 1.027 F 1.020 1.019 1.025 1.024 M 1.021 1.028 1.032 1.024 F 1.022 1.019 1.029 1.029 18 1.042 1.032 1.043 1.027 24 1.051 1.039 1.030 1.038 O Os IP o O' vNO B l O - T E 5 T 2aU*aU 30 i F. Pathologic Studies 1. T h re e -M o n th S a c rific e a. C ross Pathologic Findings G ross pathologic findings among test animals w ere not significantly different from those noted in control anim als. b. Organ Weight and Ratio Data The m ean organ weight and ratio data collected at the th re e -m o n th s a c rific e a re p re sen te d in T ab les XIII through XVIH, ' Statistically significant differences are designated by a ste risk s following the test group value. w Organ w eights, organ to body w eight and organ to b ra in (\ w e ig h t r a t i o s d i s c l o s e d s e v e r a l r a n d o m ly o c c u r r i n g i n t e r g r o u p d i f f e r e n c e s i The lack of any c o n siste n t dose re la ted re s p o n se and the absence of any deleterious histopathologic change confirm that these differences w ere not re la te d to the ingestion of A ro clo r 1242. GG 3fct 001 464 Dietary Level of A ro c lo r 1242 (ppm) Cont rol 1 10 100 a o> CT\ 01 o GO TABL E XIII T E S T M A TERIA L: A ro c lo r 1242 T w o-Y ear C hronic O ral Toxicity Study - Albino R ats O rgan W eight and Ratio Data Three-M onth Sacrifice Sum m ary of M ean Values Organ: Liver O rgan W eight (b) M alee Fem ales 21.4 13.0 22.3 12.4 23.3 14.4 25.5 ' 14.0 O rgan/B ody Weight Ratio (g /1 0 0 g) M alea Fem ales 4 . 13 4.16 4.31 3.92 4.41 4.31 4.39 O rg an /B rain W eight R atio (g/g) M ales Fem ales 10.3 6.82 10.6 6.59 11.9 7.93 15. 1 7.65 Dietary Level of A ro c lo r 1242 (ppm) C o n tro l 1 10 100 T A B L E XIV T E S T M A T ER IA L : A ro c lo r 1242 T w o -Y ear C hronic O ral Toxicity Study - Albino R ats O rgan Weight and Ratio Data Three-M onth Sacrifice ' Sum m ary of M ean Values Organ: Kidneys ( O rgan W eight (g) M ales Fem ales 4.03 2.49 4.06 2.46 4.14 2.75 4.24 2.61 O rgan/B ody Weight Ratio Ig /1 0 0 g) M ales Fem ales 0.782 0.802 0.784 0.772 0.786 0.827 0.795 0.817 O rg an /B raln W eight R atio (g/ g) M ales Fem ales 1.95 1.31 1.93 1.30 2.11 1.52 2.47 1.42 v c C t ii i i a i 0 0 465 t> L Ir tL id u a i B I 0 T E 5 T JaA oia& usi, 26 E. U rine Analyses Mean m ale and female urine analyses data are sum m arized in T ables X through XIL No differen ces w ere noted between the u rin e fro m control and te st anim als at the intervals of examination. I AOH O n a < l ) ( ) t 460 7 1 S 3 1 ` 0 M3 TABLE X T E ST M ATERIAL: A ro c lo r 1242 T w o-Y ear Chronic O ral Toxicily Study - Albino Rata Urine A nalyses Data Sum m ary of Mean Values ! D ietary Glucose Level Month: Jp p m ) S ex 3 6 9 12 18 24 Control M n n n n nn F n n n n nn M n n n n nn 100 F n n n n nn Albumin Month: 3 6 9 12 18 24 n n t t 42 43 n n n n t 42 n n t t 4 1* 44 n n t t t 43 Key: n = negative t = t r a c e ; l e e s than 30 m g / 100 m l u r in e 4 I = 30 to 100 m g / 100 m l u r in e 4 2 = 100 to 300 m g / 100 m l u r in e 4 3 = 300 to 500 m g / 100 m l u r i n e 44 = m o r e than 500 m g / 100 m l u r in e ADM 00 461 TV i s 3 ; *O 18 P W V T A B L E XI T E ST M A TERIA L: A ro c lo r 1242 T w o-Y ear C hronic O ral Toxicily Study - Albino R ats U rine A nalyses Data Sum m ary of Mean Values D ietary Level (ppm) .Control M icroscopic Elem ents Month: S e x 3 6 9 12 18 24 M N N N N N +1 F NNNNt t pit Month: 3 6 9 12 18 24 7 7 6. 7 7 6 8 8 7 8 66 M N N N N Nt 7 7 7 8 6 6- 100 F N N N N Nt 7 7 6 7 66 Key: N= N o r m a l t = trace; le ss than 5 p er low power field 1 = 5 to 10 p e r low p o w e r fie ld +2 = 10 to IS p e r low p o w e r fie ld O = m o r e th a n 15 p e r lo w p o w e r fie ld UJ o- N CO 462 * f, D ietary Level (ppm) Control 100 O O >o*T CT> Ul Oo a O' B!0 -T EST A TADLE XII T E S T M A TER IA L: A ro c lo r 1242 T w o-Y ear C hronic O ral Toxicity Study - Albino R ats U rine A nalyses Data Sum m ary of Mean Values Specific Gravity Month; S e x 3 6 9 12 M 1.029 1.022 1.033 1.027 F 1.020 1.019 1.025 1.024 M 1.021 1.028 1.032 1.024 F 1.022 1.019 1.029 1.029 18 1.042 1.032 1.043 1.027 24 1.051 1.039 1.030 1.038 nNO jftd d a lu a l B 1O * T E S T 30 f F. Pathologic Studies 1. T h r e e -M o n th S a c rific e j a. G ro ts Pathologic Findings G ross pathologic findings among test animals w ere not significantly different from those noted in control anim als. b. Organ Weight and Ratio Data The m ean organ weight and ratio data collected at the th re e -m o n th s a c rific e a re presented in T ables XIII through XVin. * Statistically significant differences are designated by a ste risk s following the test group value. O rgan w eights, organ to body w eight and organ to b ra in weight ratios disclosed several randomly occurring intergroup differences The lack of any consistent dose related resp o n se and the absence of any deleterious histopathologic change confirm that these differences w ere not re la te d to the ingestion of A ro clo r 1242. CM GG366 0 0 1 464 D ietary Level of A ro c lo r 1242 (ppm) Coht rol 1 10 100 5 O* u CTl CM TABLE XIII T E S T M ATERIA L: A ro c lo r 1242 T w o -Y ear C hronic O ral Toxicily Study - Albino R ats O rgan W eight and Ratio Data Three-M onth Sacrifice Sum m ary of M ean Values Organ; Liver O rgan W eight <g) M alea Fem ales 21.4 13.0 22.3 12.4 23.3 14.4 25.5 14.0 O rgan/B ody Weight Ratio (g /1 0 0 g) M ales Fem ales 4 . 13 4.16 4.31 3.92 4.41 4.31 4.80 4.39 O rg an /B rain W eight R atio (g/g) M ales Fem ales 10.3 6.82 10.6 6.59 11.9 7.93 15. 1 7.65 V Dietary Level of A ro c lo r 1242 (ppm) C o n tro l 1 10 100 T A B L E XIV T E S T M ATERIA L: A ro c lo r 1242 T w o -Y ear C hronic O ral Toxicity Study - Albino R ats O rgan Weight and Ratio Data Three-M onth Sacrifice ' Sum m ary of M ean V alues Organ: Kidneys O rgan W eight (g) M ales Fem ales 4.03 2.49 4.06 2.46 ` 4.14 2.75 4.24 2.61 O rgan/B ody Weight Ratio (g/100 e) M ale. Fem ale. ~ 0.782 0.802 0.784 0.772 0.786 0.827 0.795 . 0.817 O rg an /B rain W eight R atio (g/g) M ales Fem ales 1.95 1.31 1.93 1.30 2.11 1.52 2.47 1.42 V 00 465 o o f' Dietary Level of A ro c io r 1242 (ppm) Cont rol 1 10 100 C T A B L E XV T E S T M A TERIA L; A ro c io r 1242 T w o -Y ear C hronic O ral Toxicity Study - Albino R ats O rgan W eight and Ratio Data Three-M onth Sacrifice Sum m ary of M ean Values Organ: Spleen O rgan W eight (g) M ales Fem ales 0.766 0.578 0.768 0.544 0.756 0.596 0.814 0.528 O rgan/B ody Weight Ratio (g /1 0 0 g) M ales Fem ales 0.150 0.184 0. 148 0.171 0.143 0.179 0.154 0.166 O rg a n /B ra in Weight R atio (g/ g) M ales Fem ales 0.371 0.302 0.364 0.287 0. 385 . 0.328 0.468 0.288 C Ki. Ic f b99fcU'J *UV r Dietary Level of A ro c lo r 1242 (ppm) C o n tro l 1 10 100 . TA BLE XVI T E S T M A TERIA L: A ro c lo r 1242 T w o -Y ear Chronic O ral T oxicity Study - Albino R ats O rgan Weight and Ratio D ata Three-M onth Sacrifice Sum m ary of M ean Values Organ: Gonads O rgan W eight Cgl M ales Fem ales 3.60 0.114 3.51 0 . 1436 3.50 0.112 3.49 0.112 O rgan/B ody Weight Ratio (g /1 0 0 g) M ales Fem ales 0.698 0.0370 0.681 0.0452 0.666 0.0336 0.656 0.0356 O rg an /B rain W eight R atio (B/ g )* M ales Fem ales 1.74 1.66 1.78 0.0599 0.0758 0.0616 2.06 0.0619 * S tatistically significant difference at the 95 p e rc en t confidence level. O' 0 U ( 468 Dietary Level of A ro c lo r 1242 (ppm) C o n tro l 1 10 100 G <b T a\ ID *!>*3 TA B LE XVII T E S T M ATERIA L: A ro c lo r 1242 T w o-Y ear Chronic O ral Toxicily Study - Albino R ats O rgan Weight and Ratio Data Three-M onth Sacrifice Sum m ary of M ean Values Organ: H eart O rgan W eight O rgan/B ody Weight Ratio O rg an /B rain W eight R atio (g) (g /1 0 0 g) (g/g) M a le _______F e m a l e _____ _____ M a le _______F e m a l e __________ M a le _______F e m a l e 1.59 1.05 0.307 0.339 0.767 0.551 1.58 1.02 0. 307 0.320 0.752 0.540 1.55 1.10 0.295 0.331 0.788. 0.608 1.61 1.05 0.302 0.330 0.943 0.573 09tU'J ^3^ Dietary Level of A ro c lo r 1242 (ppm) C o n tro l 1 10 too i TA B LE XVIII T E S T M A T E R IA L : A r o c lo r 1Z42 T w o -Y ear C hronic O ral Toxicity Study - Albino R ats O rgan Weight and Ratio Data Three-M onth Sacrifice Sum m ary of Mean Values Organ: Brain O rgan W eight (g) M ales Fem ales O rgan/B ody Weight Ratio ( g /1 0 0 g) M ales Fem ales Total Body Weights of Sacrificed Rats (g) M ales Fem ales 2.07 2.12 1.97 1.82 1.91 1.89 1.81 1.83 0.402 0.410 0.375 0.345 0.616 0.597 0.545 0.576 , 518 520 527 534 314 318 334 319 v B 1C* - T EST oJafoa/*Ud, fa 37 c- Histopathologic Findings Histopathologic examination of tissues and organs taken f r o m a l l a n i m a l s f r o m b o th th e c o n t r o l and 100 p p m g r o u p s w a s c o n d u c te d . T a b l e s XDC and XX l i s t a l l h i s to p a th o lo g ic c h a n g e s noted. All of the lesions noted in the m icroscopic exam ination of tissu e s w ere those of spontaneous disease and a re not unusual for the albino ra t. The m o st frequent Endings w ere lesions in the trach ea and lungs, indicating chronic m urine pneumonia. These occurred in the c o n tro l as w ell as the ra ts fed A ro clo r 1242. ADH CO 0 U 471 I B 1O T E S T Ja&uiUiicd, 3c. 38 TA BLE XIX T E S T M ATERIA L: A ro c lo r 1242 T w o -Y ear C hronic O ral Toxicity Study - Albino Rats Histopathologic Changes % Three-M onth Sacrifice Group: Control N u m b er of A nim als O rean Exam ined Findines Ave raj Incidence Grade 5 M ales Trachea T rac h e itis 1 1.0 Lung Chronic resp irato ry disease 1 r.o U rinary Bladder H yperplasia 3 1.0 5 Fem ales Lung Chronic resp irato ry i disease 1 1.0 All other tissu es and organs w ere norm al histologically. Gradine System 0.5 s m inim al 1.0 = slight 2 .0 = m ild 3.0 = m oderate 4.0 = severe 5.0 = extrem e ADM C C 3 6 6 9 I 4 7 2 3 10 ' E 5 T .7/ic 39 TABLE XX .T E ST M ATERIAL: A ro c lo r 1242 T w o-Y ear C hronic O ral Toxicity Study - Albino Rats Histopathologic Changes Three-M onth Sacrifice G roup: 100 ppm N um ber of A nim als Organ Examined Findings 5 M ales Trachea T rac h e itis 5 Fem ales Lung Chronic resp irato ry disease Focal interstitial p neum onia Aver a Incidence G rad 2 1.0 2 1.0 1 * 1.0 All other tissues and organs were norm al histologically. Grading System 0 . 5 = m inim al 1.0 = slight 2.0 = mild 3.0 = m oderate 4. 0 = severe 5.0 = extrem e ACM 0 0 3 o 7Q . 473 i J hii& al B I O * T E 5 T J a k *afM i, a. 40 2. Six-M onth S acrifice a. C ross Pathologie Findings No outstanding differences w ere noted between test and control anim is. b. CTgan Weight and Organ to Body and O rgan to flrain Weight Ratio Data __------- -- T h e r e s u l t s o f t h e statistical~ an lay ses-.co fld u cted .-o n . a b --___ solute org an w eigh ts, o rg an to body w eight and o rg a n to brain weight ra tio s a re su m m arized in Tables XXI through XXVI. O rgan w eights, organ-to body weight and organ to b ra in weight ratios disclosed several random ly occurring intergroup differences. The lack of any co n sisten t dose re la ted response and the absence of any deleterious histopathologic change confirm that these differences w ere not r e la te d to the in g e stio n of A r o c lo r 1Z42. ACM 0 C 36 71 0 0 1 474 r*t 1S3' 019T nVJ7T` * Dietary Level of A ro c lo r 1242 (ppm) C o n tro l 1 10 , 100 TA BLE XXI T E S T M A T ER IA L : A ro c lo r 1242 T w o-Y ear C hronic O ral Toxicity Study - Albino R ats O rgan Weight and Ratio Data Six-Month Sacrifice Sum m ary of Mean Values Organ; Liver O rgan W eight (g) M ales Fem ales 22.9 13.1 13.8** 9.98* 22.3 22.0 13.2 11.7 O rgan/B ody W eight Ratio (g /1 0 0 g) M ales Fem ales 3.74 3.77 2.85** 4.08 3.15 4.23 4.58* 3.7 O rg an /B rain W eight R atio (g/g) - M ales Fem ales 11.7 6.86** 11.0 10.8 6.71 5.76 6.95 5 .9 4 ' S tatistically significant d ifference at the 95 p e rc e n t confidence level, i Statistically significant difference at the 99 percent confidence level; AD* 0 0 3 6 7 2 * 4- 00( 475 r JtuL,t*uiL B 1' T E S T 00 ( 476 Dietary Level of. A ro c lo r 1242 (pnm) C o n tro l 1 10 100 T A B LE XXII TEST MATERIAL: A roclor I242j 3 T w o -Y ear C hronic O ral Toxicily Study - Albino Rata |i O rgan W eight and Ratio Data <1 Six-Month Sacrifice Sum m ary of Mean Values Organ: Kidneys i O rgan W eight (g) M ale Fem ales 4,04 2.79 3.51 2,44* 3.75 3.54 2.49* 2.33* O rgan/B ody Weight Ratio (g /1 0 0 g) M ate Fem ales 0.664 0.728 0.686 0.806 0.774 0.790 i ! ! j t 0.730 0.757 O rg an /B raln W eight R atio (g/g) M ales Fem ales 2.06 1.43 1.75 1.41 1.86 1.31 1.74 1.18 * Statistically significant difference at the 95 percent confidence level. A0H 0 0 3 6 7 3 Jtv* LLV M 30 Dietary Level of A r o c lo r 1242 (ppm) Cont rol 1 10 ' 100 T A B L E XXIII T E ST M A T ER IA L : A ro c lo r 1242 T w o-Y ear C hronic O ral Toxicity Study - Albino R ats O rgan W eight and R atio Data Six-Month Sacrifice Sum m ary of Mean Values Organ: Spleen O rgan W eight (g) M ales Fem ales 0.752 0.684 0.754 0.542* 0.796 0.622 0.584 0.488* O rgan/B ody Weight Ratio (g /1 0 0 g) M ales Fem ales 0 .1 2 2 0 .1 9 6 0.154 0.172 0.142 0 .1 2 0 0.199 0.159 O rg an /B rain W eight R atio (g/g) M ales Fem ales 0.384 0.350 0.375 0.313 0.394 0 . 327 0.288 0.248 * S ta tis tic a lly sig n ifican t d iffe re n ce at the 95 p ercen t confidence level. ml B 1C T E S T I s jiW ik v u A l. a. r* I4*4 % Dietary Level of A r o c lo r 1242 (ppm) Control 1 10 100 TABLE XXIV T E S T M A T ER IA L : A r o c lo r 1242 Tw o-Y ear Chronic O ral Toxicity Study - Albino Organ Weight and Ratio Data Six-Month Sacrifice Summary of Mean Values Organ: Gonads Organ Weight (g) Males F em ales 3.55 0.0960 3.44 0.0810 3.55 0.106 3.35 0.0772 Organ/Body Weight Ratio (g/100 g) Males Females 0.593 0.0279 0.716 0.0253 0.652 0.0335 0.697 0.0251 O rgan/B rain Weight Ratio (g/g) Males F em ales 1.81 0.0492 1.71 0.0468 1.76 0.0558 1.65 ' 0.0392 O -4 CD i * Dietary Level of A r o c l o r 1242 <ppm) . _ Control 1 10 100 TA B LE XXV T E S T M ATERIA L: A ro c lo r 1242 Tw o-Y ear Chronic O ral Toxicity Study - Albino Rata Organ Weight and Ratio Data Six-Month Sacrifice Summary of Mean Values Organ: Heart u u S '- C* 1 r TEST Stic. Organ Weight (g) Mates F em ales 1.70 1.05 1.49 1.03 1.46 1.06 1.42 0.970 Organ/Body Weight Ratio ( g / 1 0 0 g) Males Fem ales 0.2R2 0.302 0.309 0.325 0.267 0.335 0.293 0.314 O rgan/B rain Weight Ratio (g/g) Males F em ales 0.867 0.538 0.741 0.595 0.722 0.558 0.700 0.492 00 f 479 u4*i r- ST Jai 0 0 C 4SO Dietary Level of A r o c l o r 1242 (ppm) Control 1 10 100 . TA BLE XXVI T E S T M A T ER IA L : A r o c l o r 1242 Tw o-Y ear Chronic Oral Toxicity Study - Albino Rats Organ Weight and Ratio Data Six-Month Sacrifice Sum m ary of Mean Values Organ; Brain Organ Weight (g) Males F em ales 1.96 1.95 2.01 1.73 2.02 1.90 2.03 1.97 Organ/Body Weight Ratio (fi/iOO g) Males Fem ales 0.320 0.560 0.415 0.546 0.369 0.597 0.416 0.635 Final Body Weights of Sacrificed Rats (y) Males F em ales 613 348 484 317 548 318 488 310 jI-. i D* O H t 4* O' B T E S T atafaud*. 47 c. Histopathologic Findings Histopathologic examination of tissues and organs taker. / f r o m a l l a n i m a l s f r o m both the c o n t r o l and 100 pp m g r o u p s was c o n d u c t e d . ; T a b les XXVII and XXVIII list all histopathologic changes noted. All of the lesions noted in the m icroscopic examination of tissu es w ere those of spontaneous disease and are not unusual for the albino rat. The m ost frequent findings were lesions in the trachea and lungs, indicating chronic murine pneumonia. These occurred in the control as well as the r a ts fed A ro c lo r 1242. *0* 003t 0 0 431 U ndttdfiiel Bl - TES T Jjo +a*uj , $4 43 TABLE XXVII T E S T MATERIAL: A r o c l o r 1242 I T w o-Y ear Chronic Oral Toxicity Study - Albino Rats Histopathologic Changes Six-Month Sacrifice Group: Control N u m b e r of Animals Organ Examined Findings Ave raj Incidence Grad< 5 Males Trachea Lung Liver Skeletal Muscle Tracheitis Focal tracheitis Focal pneumonitis Hyperemia Focal pericholangitis Focal inflammation 2 1.5 2 1.0 1 0.5 2 1*0 1 ' 1.0 1- 1.0 5 Females Trachea Lung Liver Kidney Focal tracheitis Hyperemia Hyperemia Pericholangitis Focal pneumonitis 1,, 2 t 1 1 1.0 2.5 1.0 0.5 1.0 All other tissues and organs were normal histologically. Grading System 0 . 5 = minimal 1.0 a slight 2.0 s mild 3.0 s moderate 4.0 * severe 5.0 a extreme CC3T9 AC* aduiiiiat B I O T E 5 T 49 , i TABLE XXVIS T E S T MATERIAL: A ro c lo r 1242 Tw o-Year Chronic Oral Toxicity Study - Albino Eats Histopathologic Changes Six-Month Sacrifice G ro up: 100 p p m N u m b e r of Animals Organ Examined Findings Average Incidence Grade 5 Males - Trachea Lung Tracheitis Focal tracheitis Focal pneumonitis Hyperemia 2 1.0 2 0.5 3 1.5 2 1. 5 Females Trachea Lung Focal tracheitis Focal pneumonitis Hyperemia l; 1 1- 1.0 0.5 2.0 All other tissues and organs were normal histologically. Grading System 0 . 5 = minimal 1.0 = slight 2.0 = mild 3.0 = moderate 4.0 = severe 5.0 = extreme AGH GQ368C 00( 4e3 II i:d*dt*al B l T E S T e. 50 3. T w e l v e - M o n t h S a c rific e -a--.-----G---r-o--s--s---P--a--t-h--o--l-o--g- i-e----F-in d' i n'g s / G ross pathologie findings among test animals were not significantly different from those noted in control animals. b. Organ Weight and Ratio Data The m ean organ weight and ratio data collected at the twelve-m onth sac rifice a re presented in Tables XXIX through XXXIV, Statisticallv significant differences are designated by asterisks following the test group value. f Organ weights, organ to body weight and organ.to b rain weight ratios disclosed several randomly occurring intergroup differences. * The lack of any consistent dose related response and the absence of any deleterious histopathologic change confirm that these differences were not re la ted to the ingestion of A roclor 1242. 'Of 434 1 Dietary Level (ppm) Control 1 10 100 O o .Ek oo o LH O oIOP T AnL E XXIX TEST MATERIAL: A ro c lo r 1242 Tw o-Y ear Chronic Oral Toxicity Study - Albino Rats Twelve-Month Sacrifice Organ Weight and Ratio Data Sum m ary of Mean Values Organ: Liver Organ Weight (g) Males Females 22.1 15.5 Organ/Dody Weight Ratio ( g / 1 0 0 g) Males. Females 3.79 3.52 22.7 13.7 3.45 3.33 22.4 14.4 3.48 3.44 26.3 14.4 3.78 3.59 O rgan/Brain Weight Ratio (g/g) Males F em ales 10. 8 7. 72 10.4 7. 05 10. 6 12.4 7. 39 7. 16 'tsriCjrtMyK'r r Dietary Level (ppm) Control 1 10 100 TA B LE XXX TE ST M ATERIAL: A ro c lo r 1242 T w o-Y ear Chronic O ral Toxicity Study - Albino Rats Twelve-Month Sacrifice Organ Weight and Ratio Data Summ ary of Mean Values Organ: Kidneys Organ Weight (g) Males Females 4.04 3. 19 4.31 2.93 4. 27 3.24 4.98* 3.01 Organ/Body Weight Ratio (g/100 g) Males Females 0. 692 ' 0.738 0.656 0.735 0.662 0.779 0.716 0.745 Organ/Brain Weight Ratio (g/g) Males F em ales 1.97 1.58 1.97 1. 51 2.02 2. 33 1.66 1. 50 O. O* * Statistically significant difference at the 95 percent confidence level. 00 a\ > > pr fu K d r m 1/1 a Ntn r o o j. s =i *f a rv'T'r-' Dietary Level (ppm) Control 1 10 100 TA BLE XXXI TE ST M ATERIAL: A ro c lo r 1242 T w o-Y ear Chronic O ral Toxicity Study - Albino Rats Twelve-Month Sacrifice Organ Weight and Ratio Data Summ ary of Mean Values ` Organ: Spleen Organ Weight (g) Males Females 0. 7BB 0.796 1.000 0.600 0.868 0.664 0.9&8 0. 570 O rgan/nody Weight Ratio (g/100 g) Males Females 0. 133 0. 183 0. 192 0.153 0. 136 0. 138 0. 145 O rgan/Bratn Weight Ratio (g/g) Males F em ales 0. 380 0. 396 0.458 0. 307 0.412 0. 341 0. 454 0. 285 * Statistically significant difference at the 95 percent confidence level. O nt CO r> oU* O' 0 u\n Dietary Level (ppm) Control 1 10 100 TABLE XXXII TE ST MATERIAL: A ro clo r 1242 T w o-Y ear Chronic O ral Toxicity Study - Albino Rats Twelve-Month Sacrifice Organ Weight and Ratio Data Sum m ary of Mean Values Organ: Gonads Organ Weight (g) Males Females 3.46 0. 116 3.47 0.0938 3.00 0. 141 3.67* 0. 122 Organ/Dody Weight Ratio (g /1 0 0 g) Males Females 0.596 0.0275 0. 529 0.0239 0.457 0.0332 0. 531 0.0305 O rgan/Brain Weight Ratio (g/g) Males F em ales 1.70 0.0574 1.59 0.0483 1.40 1.72 0.0719 0. 0608 * Statistically significant diffe rence at the 96 percent confidence level. vo , ? Ool>* S'tJnit'.uil. C ") * T S T yi 4* f' 1 S 3 1 * (. 0 ">( ADA* s Dietary Level (ppm) Control l 10 100 CD GO > VO > i T A B L E XXXIII TEST MATERIAL: A roclor 1242 Tw o-Y ear Chronic Oral Toxicity Study - Albino Rats Twelve-Month Sacrifice Organ Weight and Ratio Data Sum m ary of Mean Values Organ: Heart Organ Weight (g) Males Females 1.71 1.23 1.69 1.18 1.68 1.29 1.87 1.19 Organ/Body Weight Ratio f g / 1 0 0 g) Males Females 0.293 0.286 0.258 0.298 0. 263 0.312 0.268 0.298 O rgan/B rain Weight Ratio (g/g) Males F em ales 0. 835 0.776 0. 609 0. 610 0.799 0.663 0. 878 0. 591 i/ i VJt Z 5 T Jfjoy'nnfii'.u S, 490 D ietary L e v e l (ppm) Control 1 10 100 T A B L E XXXIV TEST MATERIAL: A ro c lo r 1242 Tw o-Year Chronic Oral Toxicity Study - Albino Rats Twelve-Month Sacrifice Organ Weight and Ratio Data Sum m ary of Mean Values Organ: Brain O rg a n W eight (g) Males Females 2.04 2.02 2. 19 1.95 2.11 1.95 2.14 2.00 O rg a n /B o d y W eight R atio (g /1 0 0 g) Males Females 0. 351 0. 474 0. 334 0. 516 0. 329 0.473 0. 310 0. 508 F in a l Body W eight of S acrificed A nim als (gram s) Males F em ales 584 439 660 431 642 425 698* -.409 S ta tistic a lly significant d ifferen ce at the 95 p e rc e n t confidence level. i'. ion* ^ I O - T ES T JaU ^aia^is^, 3ae. 57 c. Histopathologic Findings H isto patholo gic examination of t i s s u e s taken f r o m each r a t s a c r i f i c e d f r o m the c o n tro l and 100 ppm g ro u p s a f t e r twelve m o n t h s of testing was conducted. Tables XXXV and XXXVI list all histopathologic change noted. All of the lesions noted in the m icroscopic examination w e re those of spontaneous disea se and cannot'be re la te d to the ingestion c A ro c lo r 1242. ( i. : 0 0 * 491 B * J T S T aoo*ayu*4, jn c . 53 TABLE XXXV T E S T MATERIAL: A ro c lo r 1242 Two-Year Chronic Oral Toxicity Study - Albino Rats Histopathologic Changes Twelve-Month Sacrifice Croup: Control N u m b e r of Animals Orcan Examined Findings Incidence 5 Males Trachea Lung Urinary Bladder - Tracheitis Chronic respiratory disease Calculus 1 2 l 5 Females Trachea Lung Colon Tracheitis Chronic respiratory disease Parasites r i. i Avc Cr; 1. J. 1. 1. 1. 1. All other tissues and organs were normal histologically. C ra d irr Sv'stem 0 . 5 = minimal 1.0 = slight 2.0 = mild 3.0 = moderate 4.0 = severe 5.0 = extreme i ACM 0 0 3 6 6 9 00( 492 C6fr ' O u 6^eoo wov ta W I 59 TABLE XXXVI T E S T M A T E R IA L : A r o c l o r 1242 Tw o-Y ear Chronic O ral Toxicity Study - Albino Rats Histopathologic Changes t Twelve-Month Sacrifice Group: 100 ppm N um ber of O rgan Animals Examined Findings 5 Males Trachea Lung Liver Pancreas Kidney Pituitary Adrenal Tracheitis Chronic respiratory disease Congestion Focal atrophy Focal lymphoid infiltration Chromophobe hyperplasia Nodular hyperplasia Hematocyst Aver Incidence Gra 1 4 1 1 l 1 1 1. l.( l.< l.< f< l.( l.( l.C l.C 5 Females Trachea Lung Tracheitis Chronic respiratory disease 1 1 1. C l.C All other tissues and organs were normal histologically. Grading System 0. 5 = m inim al 1.0 = slight 2.0 s mild 3.0 s moderate 4. 0 s severe 5. 0 s extrem e. V. aoh 0C3(jo 0 0 ( 494 B I ~ - T E S T A&yi&LyuM, nc. 3 - 4. Final Sacrifice a. Gross Pathologic Findings Gross pathologie findings among test animals were not significantly different from those noted in control animals. j b. O rgan Weight ar.d Ratio Data The m ean organ weight and ratio data collected at the final sacrifice a re presented in Tables XXXVII through XLII. Statistically significant differences are designated by a s te ris k s following the te st group value. The absolute liver weight and liver to body or b ra in weight r a t i o s w e r e s i g n i f i c a n t l y e v alu a te d in f e m a le r a t s fed 100 ppm A r o c l o r 1242. Statistical evaluations of other organ weights, organ to body weight and organ .to brain weight ratios disclosed several randomly occurring intergroup dif fe re n ce s. The lack of any consistent dose re la ted re sp o n se indicates that th e s e d iff e r e n c e s w e r e not r e l a t e d to the in g e stio n of A r o c l o r 1242. ADM CC3692 T A B L E XXXVII TEST MATERIAL: A ro c lo t 1242 Two-Year Chronic Oral Toxicity Study - Albino Rata Organ Weight and Ratio Data V 1 ( r t ____________________________________ Organ Weight itary Level <g)' (ppm) M ales Fem ales Sum m ary of Mean Values Final Sacrifice I I( \ Organ: Liver I V ____________________________________________________ ____________ } Organ/Body Weight Ratio O rgan/Brain Weight Ratio (g/100 g) (g/g) M ales Fem ales M ales Fem ales C o n tro l 22.0 16.3 3.47 3.05 10. 10 8 .4 ; 1 21.9 15.7 2.93 3.00 9.97 8.03 10 22. 3 17. 1 3. 19 10. 20 8. 50 100 2 5.6 22.4** 3.95 4.02** 11.90 1 1 . 60** GO 496 * S ta tis tic a lly sig n ifica n t d iffe re n ce at the 99 percent confidence level. O - o 4 I 0 IO *I t 5 T etary Level <PPm) Control 1 10 100 TA B LE XXXVIII T E ST M ATERIAL: A ro c lo r 1242 Two-Year Chronic Oral Toxicity Study - Albino Rata Organ Weight and Ratio Data Summary of Mean Values Final Sacrifice Organ Weight (gl Males Females 5.57 3.48 6. n 3.45 Organ: Kidneys* Organ/Body Weight Ratio (c/100 g) Males Females 0.877 0.646 0. 820 0.670 5.63 5.44 3.43 3.43 0.816 0. 835 0.644 0.725 v Organ/Brain Weight Ratio (g/g) Males F emalcs 2.57 1. 79 2.78 I. 76 2.56 2.52 1.71 1.79 * O' N 6P )00 >' C00 etary Level (ppm) Control 1 10 100 o o o ID TA BLE XXXIX T E ST MATERIAL: A ro c lo r 1242 Tw o-Y ear Chronic Oral Toxicity Study - Albino Rats Organ Weight and Ratio Data Summary of Mean Values Final Sacrifice Organ: Spleen Organ Weight (g) Male Females 1. 15 0.9b 1.26 0.81 Organ/Body Weight Ratio ( g /1 0 0 g) Males Females 0. 182 0. 181 0. 169 0. 158 1.08 1.07 0. 82 0.65 0. 158 0. 165 0.153 0. 140 5 l i i i i ! Ii I J i Organ/Brain Weight Ratio (g/'g) Males Females 0.520 0.572 0. 49? 0. 145 0.495 0.499 0.409 0. 339 OUJ' etary Level (ppm) Control 1 10 100 O o> Cl * u> VO oo oUi o It*/* TABLE XL TE ST MATERIAL: A ro c lo i 1242 Two-Year Chronic Oral Toxicity Study - Albino Rats Organ Weight and Ratio Data Sum m ary of Mean Values Final Sacrifice Organ Weight Males (g) Fem ales 2.52 0. 123 . 20 0. 102 t 0. 126 Organ: Gonads Organ /Body Weight Ratio (g/100 g) Males Females 0.397 0.0222 0.424 0.0205 0.437 0.0237 3.31 0. 129 0.514 0.0278 * O r g a n / B r a i n W eig ht Rati (g/g) Males Female 1. 15 1.46 0. 062 0.0523 1.40 0.0625 1.55 0.0671 O' -ft * :tary Level (ppm) Control 1 10 100 .TABLE XL! TEST MATERIAL: A ro c lo r 1242 Tw o-Y ear Chronic Oral Toxicity Study - Albino Rats Organ Weight and Ratio Data Sum m ary of Mean Values Final Sacrifice Organ Weight Males (gV Females Organ: Heart Organ/Dody Weight Ratio tg /1 0 0 g) Males Females 1.87 2. 30* 2.08+ 2. 19* 1.54 1.5.1 1.59 1.41 0.294 0.307 0.303 0.335 0.288 0.292 0. 298 0.303 + S t a t is t ic a lly s ig n ifica n t d iffe re n c e at the 95 p e rc e n t confidence le v e l. * S ta tis tic a lly sig n ifican t d iffe re n ce at the 99 p ercen t confidence level. ADM 0 0 3 6 9 7 : i; co O m LO Organ/Brain Weight Ratio (g/gV Males Females 0. 860 1.040** 0.959* 1.010** 0.79r 0.770 0. 788 0. 74 1 OUl' obs 00 r Dietary Level (ppm) Control 1 10 100 TABLE XLII T E S T M ATERIAL: A ro c lo r 1242 T w o -Y e a r Chronic O ral Toxicity Study - Alliino Hats O rga n Weight and Ratio Data Summary of Mean Values Final Sacrifice Organ Weight (g) Males Females . 2.16 1. 95 Organ: Brain Organ/Body Weight Ratio (g/100 g) Males Female 0. 342 0. 362 Final Body Weight of Sacrificed Animals (grams) Males Females 642 551 2.20 1.96 0. 294 0. 388 759 532 2. IB 2.01 0. 313 0.379 734 547 2. 14 1.91 0. 330 0.415 652 4H5 t l t ( Ii i l l 6 t o > o o "X U1 o o oU Oo'- O' O' c. Histopathologic Findings H is to p at h o lo g ic e x a m i n a t i o n o t i s s u e s and o r g a n s ta k e n # f r o m aU a n im a ls sacrificed after 24 months os te s t was conducted. Tables XLUI through XLVI list all histopathologic change noted. Table XLVII contains the results of the histopathologic examination of the urinary bladders. Significant l i v e r in jury was found in anim als fed 100 ppm A r o c l o r 1242 (T-III). The compound a sso ciated lesions consisted of v a c u o la r change, focal hypertrophy and focal h y p e rp la sia . In addition, the w e r e l e s i o n s of in flam m atio n , n e c r o s i s , f i b r o s i s and m in o r d e g e n e ra t i o n i: t h i s g r o u p which, although s ee n in the c o n t r o l s and lo w e r t e s t g r o u p s , wer< m o re severe and frequent in the T-III group. The vacuolar change was no: seen in the control animals b u t w a s o b s e r v e d o c c a s i o n a l l y in l i v e r f r o m T - I (1 ppm) and T - H {10 ppm) a n i m a l s . The v a c u o la r change was o b s e rv e d freq u e n tly in the T -III animal* II This lesio n is morphologically indicative of fatty degeneration. Formalin-i fro zen sections of livers from representative animals which displayed j i v a c u o l a r c h a n g e s w e r e s t a i n e d with Oil R ed O to r e v e a l the p r e s e n c e of fat.i i .The v a c u o la r lesio n in the cytoplasm of these cells was positively identified as fat. The hypertrophic change found in the liver was focal and often lim ite d to the c en tral lobular a re a w here groups of cells w ere swollen to two o r three times their norm al size with clear pink homogeneous cytopl AC* OC309S 0 0 C 502 The h y p e rp la sia was associated with the sam e cells and appeared to be an extension of the hypertrophic lesion. The hyperplastic cells were also usually hypertrophic. The m ost severe examples of hyperplasia appeared a s nodular growths with limited co m p ressio n of the surrounding n o rm a l hepatic tissue. The m in o r lesions of d e gene ra tion, hepatitis, ductule cel p ro liferatio n , n e c ro sis and focal lymphoid infiltration seen in the control a n i m a l s and the T - I and T - II g r o u p s a r e l e s i o n s of spo nt an eou s d i s e a s e and a r e not r e l a t e d to the A r o d o r 1242. This level' of liv e r disease is not unusual in old anim als. * H yperplasia of the transitional epithelium was p re s e n t in the u r i n a r y b l a d d e r of anim als f r o m the c o n tro l gro up and f r o m each of the test groups. This lesion was usually associated with cystitis. All of the other lesions in the other tissues found in these animals are related to spontaneous disease and they are not unusual for r a t s of this age. Donovan E. C o r d o n , D. V. M. , Ph . D. Diplomate, A m e r ic a n College of Veterinary Pathologists AOH C 0 0 U 503 V TABLE XLIII TEST MATERIAL: A roclor 1242 Tw o-Y ear Chronic O ral Toxicity Study - Albino Rats Histopathologic Changes Final Sacrifice Group: Control Number of . A n i m a l s _________O r g a n E x a m i n e d 1I Males Heart Trachea Lung Liver Stomach Spleen Kidney Testes os 3 0 0 Prostate Adrenal Gland * > o X Brain Eye o Uoi o Findings Chronic focal myocarditis Tracheitis Chronic respiratory disease Focal degeneration Ductile hyperplasia Chronic gastritis Hemosiderosis Chronic nephritis Testicular degeneration Periarteritis Inflammation Telangiectasis Nodular hyperplasia Focal fatly change Focal encephalitis Inflammation Incidence 3 3 9 1 2 1 3 B 2 2 1 1 1 1 t l V* fus k O ni tn Average Grade 1.0 1.0 1.0 1.0 1.0 1.0 1.0 1.0 1.0 1.0 1.0 I. 0 1.0 1.0 1.0 1.0 P i; i.' f sr O' Number of Animals 14 F e m a l e s O,, 2s Ln uoi oo ip o N TABLE XLIII continued v> TE ST M ATERIA L: A r o c l o r 1242 Tw o-Year Chronic Oral Toxicity Study - Albino Rata Histopathologic Changes Final Sacrifice Group: Control Average O r g a n E x a m i n e d ____________________F i n d i n g s ___________________________ In c id e n c e _________'_____ G r a d e Heart Trachea Lung Liver Stomach Colon Spleen Uterus Kidney Ovary Chronic focal myocarditis Tracheitis Chronic respiratory disease Focal lymphoid infiltration Ductile hyperplasia Focal hepatitis Non-lipid vacuolar change Chronic gastritis Acanthosis - squamous epithelium Parasites Hemosiderosis Metritis Chronic nephritis Focal lymphoid infiltration Cyst 7 9 13 3 2 1 2 3 5 1 4 3 6 1 1 2.0 i.o 1.0 1.0 1.0 1.0 0. 5 1.0 1.0 1.0 . 1.0 2. 0 1. 0 1.0 1.0 I-1 H ? I * O 19 506 Number of Animals 14 F e m a l e s TABLE XLIII continued TE ST M ATERIAL: A ro c lo r 1242 Tw o-Year Chronic O ral Toxicity Study - Albino Rats Histopathologic Changes Final Sacrifice Group: Control * Organ Examined Adrenal Gland Salivary Gland Findings Telangiectasis Nodular hyperplasia Chronic fibrosis and atrophy incidence 9. 4 1 All other tissues and organs w ere normal histologically. 0.5 = minimal |.0 = slight Grading System 2. 0 = m ild 3.0 = moderate 4.0 = severe 5. 0 = extrem e O O A0M 0 0 3 7 0 3 Average Grade 1.0 1.0 1.0 r s Number of Animals 13 Males O <=$, I TABLE XUV T E S T M ATER IA L: A r o c lo r 1242 Tw o-Y ear Chronic O ral Toxicity Study - Albino Rats Histopathologic Changes Final Sacrifice Group: 1 ppm Organ Examined Heart Trachea Lung Liver Stomach Spleen Kidney Testes Prostate Adrenal Gland Findings Chronic focal myocarditis Tracheitis Chronic respiratory disease Focal lymphoid infiltration Cirrhosis Congestion Chronic gastritis and acanthosis Hemosiderosis Chronic nephritis. Calcification Pc ria rlc rilis Testicular degeneration Inflammation Telangiectasis Nodular hyperplasia Incidence 5 9 12 1 1 1 2 4 9 1 2 1 1 2 l f V* Is I" TO O nt in Average G rade 1.0 2. 0 1.0 1.0 1.0 1.0 1.0 2. 0 2. 0 1.0 2.0 1.0 4. 0 1.0 2. 0 I'i 507 0 T A B L E X L I V continued T E S T M A T E R I A L : A r o c l o r 1242 T w o -Y e a r C h ro n ic O ra l T o x icity Study - A lb in o Rata <> H i s t o p a t h o l o g i c C h a n g e s Final S acrifice Group: 1 ppm . Num ber of A n im a ls O rgan Examined F in d in g s 14 F e m a l e s Heart Trachea C hronci focal m yocarditis T racheitis Lung C hronic re sp ira to ry disease L iv e r D uctile hyperplasia Pancreas C hronic focal pancreatitis S tom ach C hronic g astritis U lcerative g astritis A c a n th o s is Spleen H em osiderosis K id n e y Chronic nephritis oE U te ru s Mel ritis A d ren al Gland T e la n g ie c ta s is D O ------------------------------------------------------------------------------------------------------------- o -/ Lfl OCO ( A ll other tissues and organs w ere n o rm a l h isto lo g ica lly . * Grading System Incidence 6 7 13 3 2 2 1 2 8 6 1 10 Average G ra d e 1.0 1.0 1.0 1.0 1.0 1.0 1.0 1.0 2. 0 1.0 1.0 1.0 000 509 Num ber of A n im a ls 13 M a le s o TABLE XLV T E S T M A T E R IA L : A r o c l o r 1242 Tw o-Year Chronic O ral Toxicity Study - Albino Rats Histopathologic Changes Final Sacrifice G ro u p : 10 p p m O rgan Exam ined H e a rt Trachea Lung L ive r Pancreas Sto m ach Spleen K id n e y Testes F in d in g s C h ron ic focal m yocarditis T racheitis C h ro n ic re sp ira to ry disease F o ca l hypertrophy and hyperplasia H yp e rtro p h y V a c u o la r change (fatty) D uctile hyperplasia C h ro n ic focal pancrcatits A c a n th o s is Inflam m ation, ulceration anti ncc ros is H em osiderosis Chronic nephritis T e sticu la r degeneration P e ria rte ritis Incidence 6 7 9 1 1 2 1 3 6 1 7 10 2 3 Averag G ra d l. 0 1.0 2. 0 1.0 1.0 i: o 2.0 1.0 1.0 1.0 1.0 2. 0 3. 0 3.0 -1 i. T A B L E X I,V continued T E S T M A T E R I A L : A r o c l o r 1242 T w o -Y e a r C h ro n ic O ra l T o x icity Study - A lb in o R ats H istopathologic Changes Final S acrifice G r o u p ; 10 p p m * Num ber of Average A n i m a l s _________ O r g a n E x a m i n e d _____________________ F i n d i n g s _______________________________I n c id e n c e _____________ G r a d e 13 M a le s 13 Females Prostate A d re n a l Gland Heart Trachea Lung Tongue Sto m ach ' Spleen Lym ph Node K id n e y Uterus A drenal Gland Inflam m ation T cla n g icc las is M edullary hyperplasia N odular hyperplasia C hronic focal m yocarditis T racheitis C h ro n ic re sp ira to ry disease P e ria rte ritis C h ro n ic g a s tritis and acanthosis U lceration 1I c m o s i d c r o s i s F.dcma Chronic nephritis M etritis T e la n g ie c ta s is N odular hyperplasia l 4 1 2 1 5 9 1 5 1 5 1 6 l B 1 3. 0 1.0 1.0 1.0 1.0 1.0 1.0 2. 0 1.0 1.0 2. 0 1.0 1.0 1.0 1.0 1.0 ui i-* ACM 00 0CC3707 TABLE XLVI T E S T MATERIAL: A r o c lo r 1242 T w o -Y e a r C h ro n ic O ra l T o x icity Study - A lb in o R a is H istopathologic Changes F in a l S acrifice G r o u p : 100 p p m ' Num ber of A n i m a l a _________ O r g a n E x a m i n e d _____________________ F i n d i n g s _______________________________I n c id e n c e _____ H eart Trachea Lung L iv e r Pancreas Testes Stom ach Spleen Lym ph Node K id n e y C h ron ic focal m yocarditis T rach eitis C h ro n ic re sp ira to ry disease F o ca l degeneration Focal hypertrophy H yp e rtro p h y V a c u o la r change (fatty) P e riarte ritis P e ria rte ritis C h ro n ic g a s tritis and acanthosis P e ria rte ritis H em osiderosis P e ria rte ritis C hronic nephritis 3 3 fc 1 1 1 3 1 3 1 1 1 1 4 Average G ra d e 1.0 2. 0 1.0 1.0 1.0 1.0 1.0 3. 0 2. 0 1.0 2.0 1.0 3. 0 2. 0 fr t t ( i i I I t Ml V a oo jo v 4k u*I I Num ber of A n im a ls 6 M ales 14 F e m a l e s O O o U1 Nj \ > ox o tou TADLE XLVI continued T E S T M A T E R I A L : A r o c l o r 1242 T w o -Y e a r C h ro n ic O ra l T o x icity Study - A lb in o Rats H istopathologic Changes F in al S a crifice "T O rgan Exam ined Prostate A d re n a l Gland S a liv a ry G land Eye G r o u p : 100 ppm* F in d in g s Inflam m ation F o ca l fatty change Focal chronic inflam m ation Inflam m ation Incidence 1 '1 1 1 H eart Trachea Lung L ive r C hronic focal m yocarditis T racheitis C hronic re sp ira to ry disease F o cal degeneration Focal hyperplasia Focal hypertrophy H yp e rtro p h y Nodular hyperplasia Focal chnlangicjicpatitis 4 8 6 3 2 2 3 1 1 A v e ra g e G ra d e 3.0 1.0 1.0 1.0 1.0 1.0 1.0 1.0 1.0 1.0 2. 0 1.0 1.0 ''II 1 S 2 1 * O I 3 in M UJ V uc/r:eoo wav TADLE XLVI continued T E S T M A T E R I A L : A r o c l o r 1242 # T w o -Y e a r C h ro n ic O ra l T o x icity Study - A lb in o R ats H istopathologic Changes Final Sacrifice G r o u p : 100 p p m Num ber of A n im a ls O rgan Exam ined F in d in g s 14 F e m a l e s L iv e r (continued) Pancreas Stom ach Spleen Lym ph Node K id n e y - Ute rua A d re n a l Gland Fo cal hepatitis lly p e rp la sia V a c u o la r change (fatty) C hronic focal pancreatitis C hronic g a stritis A c a n th o s is U lcerative g a stritis U lcer H em osiderosis C o n g e s tio n C hronic nephritis M etritis Telang ectasia Nodular hyperplasia A c a n th o s is A ll other tissues and organs were norm al h istologically. Incidence 1 1 11 1 2 2 1 1 4 1 7 2 8 4 1 0. 5 = m in im a l Grading System 2.0 = m ild 4.0 = severe 1 < ( j A v e ra g e G rad> 1.0 3. 0 1.0 1.0 1.0 1.0 1.0 1.0 2.0 1.0 1. 0 1.0 1.0 1.0 1.0 T -1 til X*'"- ****1 B K TEST 3* t a b l e x lv ii T EST M A T E R IA L: A ro clo rl2 4 2 T w o -Y e a r C h ro n ic O ra l T o x icity Study - A lb in o Rats H is to p a th o lo g ic Changes of the U rin a r y B la d d e r Final S acrifice G ro u p Num ber Sex Exam ined Findings* C M 11 P ro te in a c e o u s plug (34) F 14 F o c a l e p it h e lia l h y p e r p la s ia (62) T -I (1 p p m ) M 12 F 13 P ro te in a c e o u s plug (562) D iffuse ep ith elial hyperplasia associated w ith d iffu se c y s titis (566) Diffuse epithelial hyperplasia associated w ith d if fu s e c y s t it is (SHO) None T -Il (10 ppm) o x T- HI ! 100 ppm ) o Uol r\j M F M F 11 13 6 13 Diffuse epithelial hyperplasia associated with d iffu se c y s titis (651) Diffuse epithelial hyperplasia associated with diffuse cystitis and a proteinaceous plug (655) None P ro te in a c e o u s plug (744) F o c a l e p ith e lia l h y p e rp la s ia (767) : I n d i v i d u a l a n i m a l n u m b e r s a r c s h o w n in p a r e n t h e s e s . G r a d i ng S y s lc m 1 . 0 ? m in im a 1 or* trace . n - ii 11U 4.0 = severe * 'll z i*yl ri'ii w* Incidence 1 1 1 1 1 1 1 1 1 Averag G ra d e 1 2 1 3 1 t - . 2 ~j B 1O - T 5 T JzS o tA & i* a , fnc. Z yj G . Detailed D escription of Tum or Findings Tumor data for individual animals including location, weight and pathologic classification a re presented in Table XLVIII through LI. None of these tum ors could be related to the ingestion cf A ro c lc r 1242 and a r e c o n sid e re d n o r m a l for a ra n d o m population of rats this age. AOH 003713 00 515 Animal 'umbe r ml Sex J -M 2-M |6- F 47 - F 48- F 50 - F 52 - F O v ,-F cn t-* cn | - E ( f. . r' ? o o i*j ^ T I T A B L E XLVIII T E S T M A T E R I A L : A r o c l o r 1242 T w o -Y ea r Chronic Oral Toxicity Study - Albino Rats Description of T u m o r Findings Group: Control Location Abdomen Pane rea a Abdomen Lymph node Abdomen Abdomen Abdomen Abdomen Adrenal gland Abdomen A1 1-------- Weight (gram) - 11.76 64. 98 35. 09 - - - - - 88.92 n n rwi Sire (cm) 8x1x2 5x2x2 - 2x2x1 2x2x3 - -. - 13 x 11 x 4 - 11 a Cla s sificat ion Mammary fibroadenoma Adenocarcinoma Mammary fibroadenoma M ammary fibroadenoma Reticulum cell sarcoma M ammary fibroadenoma M ammary fibroadenoma M ammary fibroadenoma Mammary fibroadenoma 1`h c o c h r o m o c y l o i n a Fibroadenoma .. Vi O Months on Test 24 24 24 24 24 7 24 24 24 'i f : 517 Animal Numbe r and Sex 566 -M 569-M 604-F 609- F m 611-F Location Abdomen Spleen Abdomen Abdomen Abdomen 6 13- F Q5 15 - F O -c? --------------- Ovary Abdomen Abdome n AUM 0 0 3 7 1 5 TABLE XLIX T E S T M A T E R I A L : A r o c l o r 1242 T w o -Y ea r Chronic Oral Toxicily Study - Albino Rats Description of T u m o r Findings Group: 1 ppm Weight (grams) 15. 00 53. 50 35. 00 335.00 Size ( c m )__________ 5x3x1 5x3x1 Ixlxl 3x2x1 13 x 1 0 x 5 Classification Ade noe a rei noma Lymphosa rcoma M ammary fibroadenoma Fib ro adenoma Fibroadenoma Fib roadenoma Granulosa cell tumor Fibroadenoma Mammary fibroadenoma ur i Mont hs on Te st 24 22 24 24 24 24 23 .5 o(>j 518 Animal Numbe r and Sex 62 1- F 627-F 628-F 6 30 - M 631 -F 6 34 - F J5-F Location Abdomen Pituitary Abdomen Abdomen Uterus Abtiomen Abdomen Abdomen Pituitary AUh 0 0 3 7 1 6 TABLE X LIX continue! T E S T M A T E R I A L : A r o c l o r 1242 T w o - Y e a r C h r o n i c O r a l Toxi city Stutly - Albino Rais Description of Tum or Findings Group: 1 ppm Weight (grams) 18. 00 - 157.20 117.90 44. 00 13. 41 39. 36 Sire 1x3x3 - 6x3x3 9x6x4 7x6x2 3x2x1 6x5x3 6x5x4 Classification Fibroadenoma Chromophobe adenoma Fibroadenoma Mammary fibroadenoma M am m ary fibroa!enoma Fibroma Fib r o a de noma Fib roadenoma Fibroadenoma Chromophobc adenoma Months on Te st 24 24 24 23 24 24 24 Animal Number and Sex 6 10- M 646- M 662-M 687- F 689- F 690 - F O O -F O ai vo 1- F o Coo Ni I TABLEL TEST MATERIAL: Aroclor1242 T w o -Y ear Chronic Oral Toxicity Study - Albino Rats Description of T u m o r Findings Location Weight (grams) G r o u p : 10 p p m Size (cm) * Classification Abdomen 129.0 9x7x3 Fibroadenoma Pane reas Adrenal gland Pituitary Abdomen . - 280. 0 - - - - Islet cell adenoma Medullary tumor Chromophobe adenoma Mammary fibroadenoma Pituitary - - Chromophobe adenoma Abdomen - - Mammary adenocarcinoma Pituitary Abdomen Abdomen - 3. 90 23. i 5 - 3x3x2 7x6x3 Chromophobe adenoma Mammary fibroadenoma Mammary fibroadenoma Abdomen 175. 0 8x7x5 Mammary fibroadenoma Parathyroid gland - - Adenoma t Months on Test 24 24 24 22 24 24 24 24 24 24 Animal Number and Sex 707- F 708- F 7 1 1- F 7 12- F Location Abdomen Abdomen Abdomen Subcutaneous, abdominal TABLE L continue! T E S T M ATERIAL: A roc lor 1242 T w o -Y ea r Chronic Oral Toxicity Study - Albino Rats Description of T u m o r Findings G r o u p : 10 p p m Weight ( g r a m a ) ___ 74. 6 189.0 6. 96 155. 0 Size (cm) 7x6x3 5x4x1 3x2x1 11x8x3 Classification Mammary fibroadenoma Mammary fibroadenoma Mammary fibroadenoma Fatty tissue fibroma o o ui K> o Oi OOOJ ,, k Months on Te at 24 24 24 24 (O 310-TEST Animal Numbe r and Sex 74 4 - M 775- F 785-F 789 -F m f 94-F Location Pane reas Ovary Pituitary Adrenal gland Abdomen TABLE LI T E S T M A T E R I A L : A r o c l o r 1242 T w o -Y e a r Chronic O ral Toxicity Study - Albino Rats Description of T u m o r Findings G r o u p : 100 p p m Weight (grams) 260. 0 Si z.c (cm) - 10 x 10 x 5 Classification Islet cell adenoma Granulosa cell tumor Chromophobe adenoma Medullary tumor Mammary fibroadenoma Months on Teat 24 24 24 24 23 OU en ro / Monsanto H*rn* ol No m int* C o m t i n r u m l or S l i M 0 P t. O m llO ft Med. & Fnv. H l t h ^ P o i u . o * T<li i-iury G n a t Toxicolocv Manacer N /IS M M A C I 19 6 w tin .il Cl O u t A n n u l i SUnr S 31 . 800 C llA D B l 'l rnn| LKihOn Creve Co 0*1 o ' u in in c iu M 2/1/76 D i L_i n f c m p i o y R - Typ oi A m ia t* t w o u i f o ry f r o m in o p p o m * p o i of i n n tnc *>ncn m o i l i p p r o p o n t i y g n u i o n tn coa* d iow.j , m a ntr in u it q o r A m>0 O l 0 iy Technical ^ Accomplishment of significant results. O c/.0 in cniAvamanl *n a Ul itin iticjn e* to M o n u m o w i C tr o ry Co m s' At a recent meeting in Creve Coeur, Glenn Schweitzer, head of the OfficToxic Substances of EPA, offered some interesting comments. He observe that Monsanto's toxicologists were held in high regard at EPA. Howeve since it lacked an in-house toxicology facility, Monsanto as a company not as highly regarded overall as duPont, Carbide, or Dow. Dr. Wright's professional and personal characteristics have contribute nificantly to Monsanto's image at EPA. He has shown unusual persevera. and dedication, frequently involving his own time, to review and interp large volumes of data which he subsequently organized for presentation EPA officials. Particularly noteworthy weri his efforts on polvchlorin biphenyls (Aroclors) and chlorinated isocyanurates (ACL products). In former instance, his excellent analysis and synthesis of widely scatter observations played a prominent role in forestalling EPA's promulgation unrealistic regulations to limit discharges of polychlorinated biphenyl EPA's proposed regulations would have precluded the use of these materi by Monsanto's customers. With respect to chlorinated isocyanurates, Dr. Wright has had several contacts with individual scientists at EPA t 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 played, he has made an important contribution to Monsanto's image at El That favorable image will become increasingly important to Monsanto as areas of interaction with EPA multiply.