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I '1 TOWOLDMONOOQ1748 Sequence of Tables 1. The Mortality, Length of Survival, end Changes in Weight of Aniaals Exposed to the Vapor of Aroclor 1242 In Air* 2* The Mortality, Length of Survive!, end Changes in Weight of Aniaals Exposed to tha Vapor of Aroelor 1254 in Air. 3* The Mortality, Length of Survival, and Changes in Weight of Exposed to Conditioned Air* 4r Suaaary of the Data on tha Mortality Aaong Experimental and Control Aniaals . 5* The Average Changes in Weight of the Survivors Among the Original Croups* . GBRN002715 TOWOLDMONOOOI749 *I1. ii Sequence of Figures 1. Weight of Cat A-499 Recorded at Weekly laterals During tha Period of Exposure to tbe Vapor of Aroelor 1242 in Air - 1.9 aicrograms per liter (Scperimeat No. 3). . 2. Weights of Individual Guinea Pigs B-599, B-800, B-8oi aad 3-828 Recorded at Weekly Intervals Duriag the Period of Sxposure to .the Vapor of Aroelor 1242 ia Air - 1.9 micro grams per liter (Experiment No. 3), 3* Weights of Individual Guinea Pigs B-802, 3-803 aad B-8o4 Recorded at Weekly Intervals Duriag the Period of Sspo- sure to the Vapor of Aroelor 1242 ia Air - 1*9 mierograms per liter (Experiment No. 3) * 4. Average aad Baage of Weight Recorded at Weekly Intervals of Gulaea Pigs 3-599 end 3-801 to B-8o4 that Survived throughout the Period of Exposure to the Vapor of Aroelor 1242 ia Air - 1.9 aierogreas per liter (Experiment No. 3). 5. Weights of Individual Vice 3-587. 3-588, 3-589 and 3-800 Recorded at Weekly Intervals Duriag the Period of Expo- sure to the Vapor of Aroolor 1242 ia Air - 1.9 aicrograas , per liter (Experiment No. 3). 8. Weights of Individual Mice B-570, S-571, B-572 aad B-921 Reeorded at Weekly Intervals Duriag the Period- Of Scposure to the Vapor of Aroelor 1242 ia Air - 1.9 mierograss per liter (Experiment No. 3) 7. Weights of Individual Mice 3-573, B-574-, B-575 , 3-578 aad .B-801 Recorded at V7eekly Intervals Duriag the Period of Scposure to the Vaoor of Aroelor 1242 ia Air - 1.9 micro grams per liter (Szperissat No. 3). 8. Average and Range' of Weight Reeorded at Weekly Intervals of Mice B-587, B-568, B-571, B-572, B-573 aad 3-574- that Survived throughout the Period of Exposure to tha Vapor of Aroelor 1242 la Air - 1.9 mierograss per liter (Ex pendcent No. 3) - G GBRN002716 TOWOLDMONOOOI750 ill 9* Weights of Individual Rabbits H-106l, 2-1062, j-784, 7-828, 7-S6l, 7-863, 7-864 end- 7-886 Recorded at Weekly Intervals During the Period of toosure to the . T(ESxpre_r?imf eAntroNcolo. r31)2.42 in Air- - 1.9 aicrograms 'per liter 10 Weights 0f Individual Rabbits H-1063, 2-1064, 7-84, ' 7-324, 7-7*0, 7-823* 7-862, 7-285 end 7-837 Recorded at Weekly Intervals During the Period of Exposure to the Vapor of drooler 1242 in Air - 1.9 micrograns psr liter (Experiment No. 3) 11. Weights of Individual Rats L-992, L-993 and L-994 Re corded at Weekly Intervals During ths Period of Exposure to the Vapor of Aroclor 1242 in Air - 1.9 aicrograms per liter ( Experiment No. 3). . 12. Weights of Individual Rats 1-995* 1-996 and 1-997- Re corded at Weekly Intervals During the Period of Exposure to the Vapor of Aroclor 1242 in Air - 1*9 oicrograms per liter (Experiment No. 3) ' 13. Weights of Individual Rats 1-993, 1-999* 1-1000, L'-l and . M-445 Re corded at Weekly Intervals During the Period of Exposure to the Vapor of Aroclor 1242 in Air - 1.9 micro grams per liter (Experiment No. 3) 14. Average and Range of Weight Recorded at Weekly Intervals of Rats 1-992 to 1-1000 that Survived throughout the Period of Exposure to ths Vapor of Aroclor in Air - 1.9 aiorograss per liter (Experiment No. 3)* 15. Weight of Cat A-500 Recorded at Weekly Intervals During the Period of Exposure to the Vaoor of Aroclor 1254 in ' Air. - 1*5 microgrecs per liter (Experiment No. 2) 16. Weights of Individual Guinea Pigs B-605, B-6c6, B-607, B-o72 and 9-799 Recorded at Weekly Intervals During the Period of Exposure to the vapor of Aroclor 1254 in Air - A 1.5 micrograms per liter (Experiment No. 2) . 17. Weights of Individual Guinea Pigs B-6c8, B-609 and B-6l0 Recorded at Weekly Intervals During ths Period of Expo sure to the Vapor of Aroclor 1254 in Air - 1.5 micrograms G per liter (^periment No. 2). GBRN002717 TOWOLDMONOOQ1751 1$ Iwiji and Range of Weight Recorded at Weekly Intervals of &ulaaa Pigs B-606 and B-606 to B-6l0 that Survived throughout tha Period of Exposure to the Vapor of Aroelor 1254 la Air - 1.5 micrograss per liter (Experiment No. 2). 19. Weights of ladiTidual Mice 3-577* B-576 ead B-579 Recorded at Yfeekly Intervals Ourlag the Period of Exposure to the Vapor of Aroelor 1254 in Air - 1.5 aicrograas per liter ( Experiment No 2) 20. Weights of'Individual Nice B-56O. B-561, 3-562 and B-919 Recorded at Weekly Intervals During the Period of Expo sure to the Vapor of Aroelor 1254 in Air - 1.5 micrograms per liter (Sxperiaaat No 2) 21. Weights of ladividusLl Nice B-563, B-564. 3-565, B-566, B-I02. B-0O3,. 3-o73 emd' 3-663 Recorded at Weekly Inter vals During the Period of Exposure to the Vapor of Aroolor 1254 in Air - 1#5 aicrograas per liter (Experiment No. 2). 22. Average sad Raage of Weight Recorded at Weekly Intervals of Nice B-577 to B-560, 3-562 ead 3-565 that Survived throughout the Period of Exposure to the Vapor of Aroolor 125*r in Air - 1.5 micrograas per liter (Experiment No. 2). 23* Weights of Individual Rabbits H-IO65 and 3-1066 Recorded - at Weekly Intervals During the Period of Exposure to the Vapor of Aroelor 1254 in Air - 1.5 aicrogreas per liter (Experiment No. 2). 24. Weights of Individual RabbitsH-1067: and 3-1066 Recorded at Weekly Intervals During the Period of Exposure to the Vapor of Aroolor 1254 la Air - 1.5 aicrograas per liter (Sxperiaaat No. 2) . . 25. Average ead Range of Weight Recorded at Weekly Intervals of Rabbits S-1065 to 3-1066 that Survived throughout the Period of Exposure to the Vapor of Aroelor 1254 in Air *1.5 aicrograas per liter (2^?crimen: No. 2). 26. Weights of Individual Rats M-2, tf-3 and 11-4 Recorded at Weekly Intervals Duriag the Period of Exposure to the Vapor of Aroelor 1254 in Air - 1.5 micrograas per liter (Sxperiaaat No. 2). 6BRN002718 TOWOLDMONOOOI752 27 . Weights of Individual Rats M-5. N-6f K-7 end M-g Re corded at Weekly Intervals During the'Period of Srsosure to the Vapor of Aroelor 1254- in Air -1.5 mioregraiss per liter (Experiment No* 2) - 2i Weights of Individuel Rats U-9, 15-10, H-ll end 15-140. Re corded at Weekly Intervals luring the Period of Exposure to the Vapor of Aroelor 1254* in Air - 1.5 microgracs per liter (Experiment No. 2) -* 29* Average and Range of Weight Recorded at Weekly Intervals of Rats 15-2 to 11-11 that Survived throughout the Period of Exposure to the Vapor of Aroelor 1254. in Air - 1.5 mierograas per liter (Experiment No. 2] 50. Weight of Control Cat A-510 Recorded at Weekly Intervals During the Period of Confinement in Conditioned Air (Ex periment No. 2) 31. Weights of Individual Control Guinea Pigs B-6ll, S-&L2, B-613 and 3-79g Recorded at Weekly Intervals During the Period of Confinement in Conditioned Air - (Experiment No. 2). 32. Weights of Individual Control Guinea Pigs S-6l4>, B-615 and S-6l6 Recorded at Weekly Intervals During the Period ' of Confinement in Conditioned Air (Experiment No. 2) 33. Average end Range of Weight Recorded at Weekly Intervals of Control Guinea Pigs B-6l2 to B-616 that Survived throughout the Period of Confinement in Conditioned Air (Experiment No. 2). 3^. Weights of Individual Control Nice B-567* B-5gg B-5S9 ' B-9oS and 3-969 Recorded at Weekly Intervals During the Period of Confinement in Conditioned Air (Experiment No. 2). 35. Weights of Individual Control Mica B-590, B-591, B-592, B-S64-, S-936 and B-967 Reoordad at Weekly Intervals Dur- lag the Period of Confinement in Conditi oned Air (Experi ment No. 2) '. 36. Weights of Individual Control Nice 3-593 B-594-, 3-595 B-590, 3-933, B-935 and B-970 Recorded at Weekly Intervals During the Period of Confinement in Conditioned Air (Ex periment No. 2). EBRN002719 TOWOLDMONOOOI753 vi 37. Average aad Range of Weight Recorded at Weekly Intervals of Control Mice 3-566, B-569, 3-591 and 3-591+ to 3-596 that Survived throughout the Period of Confinement of Conditioned Air ( 9efinest No. 2). 36. Weights of Individual Control Rabbits a-1069 aad S-1070 Recorded at Weekly Intervals During the Period of Con finement in Conditioned Air (Experiment No. 2}. 39. Weights of Individual Control Rabbits E-1071, Si-1072, 7-133 end 7-202 Recorded at Weekly Intervals During the Period of Confinenent in Conditioned Air (a^eriaant No. 2) 40. Average aad Range of Weight Recorded at Weekly Intervals of Control Rabbits E-IO69. S-1070, 7-163 and 7-202 During the Period of Confinenent in Conditioned Air (xbraeriaeat No. 2). ' 4-1. Weights of Individual Control Rats tf-12, M-13 and M-14 Recorded at Weekly Intervals During the Period of Con-, flneaent in Conditioned Air ( Experiment No. 2) 42. Weights of Individual Control Rats il-15, M-l6 and M-17 . Recorded at Weekly Intervals During the Period of Con . flneaent in Conditioned Air (Experiment No. 2). 43. Weights of Individual Control Rats lf-l6, lf-19, 11-20, M-21 and M-351 Recorded at Weekly Intervals During the Period of Confinenent in Conditioned Air (Experiment No. 2). 44. Average and Range of Weight Recorded at Weekly Intervals of Control Rats U-12 to M-lS, U-20, M-21 aad M-531 that Survived throughout the Period of Confinenent In Con ' ditioaed Air (Experiment No. 2). 6BRN002720 TOWOLDMONOOOI754  The Toxicity of the Taper of Arotuor i242 ana of Aroclor 1254 (Supplement to Heport or June 22, 1955) Scone This supplemental report presents the results obtained when separate groups of animals were exposed, respectively, to Aroclcr 1242 end Aroclor 1254 in lower concentrations in the atmosphere and for a longer period than were those employed in the experiments described in the report of June 22,. 1955* Since these experiments involved a different period of time and a dif ferent season of the year, a new group of control aniaals was included. . Summary of Results 1. In an experiment in which, for 7 tours per day on 150 days over a period of 214 days, aniaals were ess os el to air bearing Aroolor 1242 in the concentration of 1.9 micro grams per liter (O.li ppm), the incidence of mortality among the various species, with the exception of the rabbits (which died during an epidaaio of pulmonary disease), was no greater than that encountered a similarly constituted control group. The growth of the experimental animals was unaffected. Gbrhoo272i TOWOLDMONOOOI755 No gross or microscopic evidence of general or specific in jury was found in the tissues of the animals as a response to their prolonged exposure to Aroolor 1242. 2. Zn another experiment, in which animals, were exposed to air hearing Aroolor 1254 in the ooneentration of 1.5 micro grass per liter' (0*11 ppm), for 7 hours per day on 150 days ever a period of 213 days, the incidence of mortality was no greater than that observed among corresponding groups of oontrels. The growth of the experimental groups did net differ significantly from that of the controls*. No relevant gross pathological changes were observed in the animals, but micro scopic lesions of a mild, nonspecific, toxic type were found in the livers of some of ths guinea pigs and mice end in all of the rabbits and rats* Similar changes were also found in ths renal tubules of the rats* Although microscopic abnormali ties of this type are often found in the tissues of "normal" animals, those observed in certain of the control animals were ,, not as severe as they were found to be in the corresponding species of animals that had been exposed to Aroclor 1254* The conclusion seems to be warranted, therefore, that prolonged respiratory exposure to Aroclor 1254 is capable of causing some injury to ths tissuss of susceptible animals, under conditions in whieh the atmospheric concentration of the material is of the order of 0*11 parts per million* GBRNQ02722 TOWOLDMONOOOI756 Sreosure to the Veoore T2ie method. of volatilization of either of the Aroclors froa a heated glass well was sinilar to that described in the earlier report (Tune 22, 1955), except that i3 these experi ments the temperatures of the Aroelors la their respective wells were decreased and the rates of air-flow over the surface of tba . liquids were increased, la the ease of Aroelor 1242, which was heated to 55* to 6o*C, the air passed ever the liquid at the rate of $00 liters per minute as measured by a venturi-meter attached to an inclined manometer. Aroelor 1254 was maintained between 115* end 125*0 while 700 liters of air per minute passed over its surface before entering the chamber. During a 7-hour period oa each of 5 days per week, 3 groups of animals (2 experimental and 1 control) were confined simul taneously in seperate 600 liter chambers as described earlier. Zach group consisted originally of 1 eat, 6 guinea pigs, 10 mice, 4 rabbits and 10 rats. When an animal in eny of these , groups died, it was replaced soon thereafter. The ooatent of Aroelor in the air was determined daily by the turbldimetric 'method described ia the previous report. In these experiments samples were collected by passing air at the rate of 1 liter per minute for 4 minutes through a Willson GBRN002723 TOWOLDMONOOQ1757 combustion furnace and then through 2 midget bubblers in series, each containing 10 ml of 0.1 N sodium hydroxide. Sraeriaental Results . Aroclor 3.24a. Srseriaent No. me pertinent data relat ing to the fate of the individual smimals of this experiment are . detailed in Table 1 and summarized in Table 4. One oat, 5 guinea pigs, 6 mice and 9 rats survived following their subjection for 7 hours on each of 150 days (over 214 days) to air bearing the vapor of Aroclor 1242 in the concentration of 1.9 microgrems per liter (O.ld ppm). Sevan additional animals, introduced as sub stitutes for animals that had died during the experimental per iod, were alive at the conclusion of the final period of expo sure; these are listed to show their species and, in parenthe ses, the number of periods of exposure to which each was sub jected is given, as follows: 1 guinea pig (2o), 2 mice (45, 135) 3 sabbits (S, 13 end 34), and 1 rat (107), ,, In the initial group, 10 fatalities occurred as follows: 1 guinea pig after 124 periods of exposure; 4 mice after 14, 14, 102 and 150 periods, respectively; 4 rabbits after 17, lid* lid and 129 periods, respectively; and 1 rat after 43 periods. In addition, a substituted mouse died after 135 periods of GBRN002724 TOWOLDMONOOOI758 5 exposure, 11 substituted rabbits died, for the asst part, during the first 2 weebs after they bad been' introduced. Ten of tbs 15 rabbits died within a period of 3 weebs during a leTera epidastio of pneumonia* . Except in tbe ease of tbe rabbits, tbe incidence of mor tality was no greater enoag tbe experimental animals than among tbs oontrols (Tables 3 and b) Among tbe original group of controls 1 eat, 5 guinea pigs,, b mice, 2 rabbits and 9 rats survived throughout tbe entire period of tbeir confinement, for 7 hours on each of 150 days over 213 days in a chamber supplied with conditioned air* Three other animals, 1 rabbit end 2 mice, were billed after 3b, 109 and 113 periods of con finement, respectively, in a chamber in which air containing ' no Aroclor was conditioned with respeot to dust,humidity and temperature* Other control animals, introduced as sub stitutes for those that bad died or been billed, survived dtiying the indicated numbers of periods of confinement, re spectively: 1 guinea pig, 33; 6 mice, 3, 3, 13, 37, bl and 66; 2 rabbits, 113 and 119; and 1 rat, 113* Seven fatalities occurred among the. original group as follows: 1 guinea pig after 9b periods of confinement; b mice after b, 112,' 137 and Ibl periods, respectively; 1 rabbit after 30 periods, and 1 rat after 29* Tro mice used as substitutes died after b and 33 periods of confinement, respectively* EBRN002725 TOWOLDMONOOOI759 la summary, in each of tbs original (experimental and con trol) groups, 1 guinea pig, 4 sloe and 1 rat* died* No specifio signs of intoxication were observed in the members of either the experimental or control groups. . The weights, observed at weekly intervals, of tbs individ ual animals (cats, guinea pigs, mice, rabbits end rats-) exposed to Arodor 1245 (Experiment No. 3) are shown in Figures 1 to 3, 5 to 7 end 9 to 14. The average weight of the original groups end the greatest and least weights attained by individuals within these groups of guinea pigs, alee and rats that survived are plotted in figures 4-, 3 end 14-, respectively. Compares la * data for the controls are to be found in Figures 30 to 44. The av erage changes in weight of the various survivors of the original groups (experimental and control) are shown in Table 3* The ap plication of the widely used "tn test indicates that the growth of the enimals exposed to Aroclor 1242 did not differ signifi cantly from that of the control group. . All of the animals were examined post-mertem, and the vis cera of nost of them were examined microscopically. Among the exposed group, 1 guinea pig, 3 aice and 13 rabbits that died, and 1 rat that was killed when moribund, were victims of in cidental infeotious disease. The eat, guinea pigs, nice and rats that survived bad normal viscera. The 3 rabbits used as 6BRN0D2726 TOWOLDMONOOOI760 replacements survived. Two of them were found to have hepatic lesions of eoccidiosis aad the other had no gross sicrosoopic abnormalities. The control anisals that died were found to baTe pneumonia. Of those that survived, 1 rat, 2 guinea pigs and 1 mouse had focal or diffuse ejtoplasoio vacuolatioa of the hepatic cells, the etiology of which was not apparent. The remaining animals that survived had. essentially normal viscera. Aroclor 1254. Sroerimeat Wo. 2. The pertinent-data relat ing to the changes in weight and fate of the Individual animals subjected to prolonged lnternittant exposure to the vapor of Aroclor 1254 are shown la Table 2 and summarized In Table 4-. One cat, '4 guinea pigs, 6 mice, 4 rabbits and 9 rats from the . original group survived following their exposure for 7 hours on each of 150 days over a period of 213 days to air containing &rodor 1254- In the concentration of 1.5 micrograms per liter (0.11 ppm). One guinea pig and 1 rat from the original group were hilled after 30 and 149 periods of exposure, respectively. Seven animals Introduced into the group latar survived. They are listed by species aad periods of exposure, in parentheses, as follows: 2 guinea pigs (49 aad 120)j 4 mice (49, 66, 115 end 135) J 1 fat (139). Among the original group only 4 mice and 1 guinea pig died after 12 to 101 periods of exposure, and 6BRN002727 TOWOLDMONOOQ1761 e souse, which, was & replacement, died after 1d periods of ex posure, the incidence of fatalities aaong then being slightly less than that encountered eaong the controls* No general or specific signs of intoxication were noted esocg the ea^erimsntal Calais during or after their exposure. The weights, at weekly intervals, of the individual animals (i.e., cat, guinea pigs, aloe, rabbits and rats) exposed to Arodor 1254 are presented in Figures 15 to 17, 19 to 21, 23 and 24, and 26 to 2d* The average weight of the original groups, end the greatest and least weights attained by individuals within these groups of guinea pigs, nice, rabbits end rats that survived are presented graphically in Figures Id, 22, 23 and 29, respec tively* The rates of growth of the experimental groups did not differ significantly from those of the control group (Table 5) All of the animals were examined post-mortem, and the vis cera of most of them were examined microscopically. The guinea pig that died exhibited chronic pyelonephritis, pulmonary hyper emia and edema, and degenerative lesions in the brain and liver. The visceral lesions were related, no doubt, to the renal in fection* Tha deaths of the 4 mice were attributable to acute bronchitis and pneumonia* The viscera of the cat that-survived were normal* Of 7 guinea pigs that were living when the exper iment was terminated, 3 had normal viscera end four had slight 6BRN002728 TOWOLDMONOOOI762 alterations of hepatic calls characterized by cytoplasmic vacu- olation. Ten nice survived and of these 6 had nozsal viscera and 4* had slight degenerative changes in the liver. ?our rab bits hilled 1 to 15 days after the last period of exposure had diffuse hepatic degeneration. The character of the lesions var ied fsoa cloudy, hyalins or hydropic degeneration and. Included varying degrees of fatty metamorphosis. The other viscera of these aninals rare normal. Ill of the rats were examined and found to have degenerative lesions of the liver of-slight to moderate degree. The lesions of greatest severity were'found in the rat that was hilled and examined on the first day .after the last period of exposure. Two rats had chronic pyelonephri tis, end the remainder had slight degeneration of the renal tubules. . . Pis cuss ion The preceding report (dated June 22, 1955) indicated that no harm resulted to animals that inhaled Aroolor 124-2, inter mittently, over periods not exceeding 4- months when the con centration in the' atmosphere approached that of saturation. The present report demonstrates, in terms of rates of growth, incidence of mortality and non-occurrence of pathologic changes, Gb^0q2729 TOWOLDMONOOOI763 ft - 10 - that no injurious effects resulted fraa more prolonged, later* aitteat exposure (more then 6 months) to tbs- coaceatretioa of O.lS ppa lathe atmosphere (1.9 mierogrems per liter). This concentration Is nearly twice that (1 alerograa per liter) . re cocaended by the Acer! can Ocnfereace of Goverareata1 Hygien ists (Arch, lad. Hyg. Occup. Med., 2:530, 19540 for a ahloriaated diphenyl of unstated chlorlas content. The exposure of anlsals to Aroclor 1254- failed to induce haraful effects la the fors of retardation of growth, or of mortality, but histopathologic evidence of appereatly reversible hepatic cellular injury was found ia the animals* These find ings cannot certainly be attributed to the effects of Aroclor 1254> because of the appreciable incidence of pneumonia among both experimental and control animals. When these nonspecific toxic changes in the viscera of the animals were associated with pneumonia, they were readily explained thereby, but they were also found in animals that had been exposed to the vapor of Aroclor 1254* and were free of pneumonia. That these may have represented toxio effects of exposure to Aroclor 1254- fiads support in the fact that the livers of the exposed rats (report of June 22, 1955) ware significantly heavier in relation to their body weight, than were those of control rats. It would appear that this material, which is reported to contain 5^ GBRN002730 TOWOLDMONOOOI764 per cent of chlorine, is somewhat aero tozie than is Arbeler 1242, which contains only 42 per cent of chlorine. C.S. Drink! proposed (J. lad. 2rg. Toxicol., 21:155, 1939) as a threshold concentration for a sasple of a chlorinated diphenyl that con tained 50 *3 55 P- cent of chlorine, the value of 0.5 micro gram per liter, which is about one-third of the concentration of Aroclor 1254 to which the aniaals were exposed in this ex periment. Proa The Kettering Laboratory in the Oepartaent of'Pre ventive Medicine and Industrial Health, College of Medicine, University, of Oincianati, Cincinnati, Ohio. Project No* 47* Experimental Work and Report by: Joseph P Treoa, Ph.D.. ' Prank ?. Cleveland, K.2. Prederic S. Shaffer, M.S. John Oappel, B.S. Robert A. Boiler, M.S. John P. Torbeek, B.S. June 2S, 1955 aobert a. aenoe, sui). 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