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Clvo Institutes TNO Report NO. V 83.285/291099 LIFESPAN ORAL CARCINOGENICITY STUDY OF VINYL CHLORIDE IN RATS (Final Report) # A/I, ; #- ' civo institutes tno nethertands organization for applied acientific research * GCD dhrtslon tor nutrition and lood rtttch tna P.o. box 360 3700 AJ rets notheHindi Report No. V 83.285/291099 LIFESPAN ORAL CARCINOGENICITY STUDY OF VINYL CHLORIDE IN RATS (Final report) Authori : Dr H.P. Til H.R. Imael Dr V.J. Feron At the request of : Verband Kunststofferzcugende Industrie E.V., Frankfurt am Main, B.R.D. Project number : E 79-1099 Start of the study : August 21, 1979 Termination of the study: June 29, 1982 Study director : Dr H.P. Til Study supervisor : Dr V.J. Feron Approved, by : Dr A.P. de Croot Date : September, 1983 Total or partial publication of thi* report without writton aaaant u not >llow<fl 73**10 coo-otei V 83.285 CONTEXTS 6. REFERENCES 7. AUTHENTICATION 8. RETENTION OF RECORDS AND SPECIMENS STATEMENT OF CLP COMPLIANCE TABLES FIGURES ANNEXES 3- 28 30 31 32 33 70 71 R&S 134162 V 83*285 r SUMMARY 4- X. The oral carcinogenicity of vinyl chloride monomer (VCH) vit examined in a lifeapan study (149 weeks) with five groups of tfistar rats, each consisting of 100 males and 100 fesiales, except for the top-dose group which comprised 50 males and 50 females. VCH was administered by in corporating polyvinyl chloride (PVC) powder with a high VCM content into the diet. The diet was provided daily for a period of A consecu tive hours, whereas food was withdrawn during the other 20 hours. The use of this way of oral VCM administration resulted in the following exposure levels: 0 (control), 0.014, 0.13 and 1.3 mg VQi/kg body welght/day. An extra control group_.of 100 rats/sex was housed in a separate room. Additional satellite groups Of 1* male and 10 female rats, each re ceiving the same treatment as the main groups were used for determi nations of glutathione levels in the liver after 9 end 18 months.. Observations were made of general appearance mortality, growth, food intake, thrombocyte count, prothrombin time, glutathione levels in the liver, gross pathology and microscopic pathology of the liver and of all grossly visible tumours or presumable tumours in the abdominal cavity, the glands of Zymbal and the mammary glanda. 2. General health, behaviour, body weight and food Intaka were not ad versely affected by the test Substance. 3. In the second half of the experimental period, mortality in the extra control group was higher than in all other groups. This wes most prob ably due to a high Incidence of chronic respiratory disease in the extra control group. In the final stage of the study, the mortality in Che top-dose group was slightly higher than In the lower dose groups and the controls. 4. Thrombocyte count, prothrombin time aDd liver glutathione levels did not show treatment--related differences among the groups. T* R&S 134164 V 83.285 ^ - 5- 5. A clearly higher Incidence of grosaly visible, tumourous, liver nodulea warn fouiid in both male* and females of the top--dose group than In any of the other group*. Moreover, in females of the top-doae group the incidence of hepatic cyatc was considerably higher chan in con trols. 8, Microscopic examination of the liver revealed increased incidences of liver-cell polymorphism hepatic cysts, foci of cellular alteration, neoplastic nodules and hepatocellular carcinomas in the top-dose group as compered to the control group. Moreover, a hepatic angiosarcoma was found in one male and two females of the top-dose group, whereas no aucb tumours were encountered In any of the other groups. The number of animals hearing foci of Cellular alteration in the liver was also statistically significantly increased in females of the aid-dose group as compared to controls. In addition, in females but not in msles, the incidence of basophilic foci of cellular alteration in the liver was Statistically significantly higher in both the lovand the mid-dose group than in the control group. 7: There was so evidence of VQf-feedlhg effecting the incidence of ab dominal mesotheliomas or the type and incidence of mammary gland tu mours. No Zymbal gland tumour was found. 8, It wss concluded that under the conditions of the present experiment: - VCH st s level of 1.3 ag/kg body welght/day induces neoplastic and non-neoplastic changes in the liver of rsts, - VCH at a level of 0.13 mg/kg body wclght/day may lead to more female rata bearing foci of cellular alteration in the liver, - VCM et levels of 0.01* or 0.13 mg/kg body welght/day may result in an increased incidence of basophilic foci of cellular alteration in the liver of female rats, - 0.13 mg VQ</kg body welght/day la s "no-obaerved-adverse-effectlevel* with respect to the induction of tumours in rats. V 83.285 t. w -6- 9. Risk estimation based on the results of the present rat study and taking into account the prudence of the linear model applied and a lesser sensitivity of humans to the carcinogenic action of VCM in comparison with rats, indicates chat the cancer risk of a likely maximum oral dally intake of 0.1 pg VCM per person per day cag be practically neglected. 00 So (n 134165 'V V 63.285 LIFE-SPAN ORAL CARCINOGEN'!CITY STUDY OF VINYL CHLORIDE IN RATS 1. INTRODUCTION From a previous life-span oral toxicity study of vinyl chloride monooar (VCM) in rats with dose levels of 1.7, 5.0 and 14.1 mg/kg body welght/day (Feron et si., 1978; 1981), it appeared that hepatic angiosarcomas occurred at dose levels of 5.0 og/kg body veight/day and above, while hepatocellular tumours (neoplastic nodules and carcinomas) were found at all dose levels. Zymbal gland tumours were found at VCM levels of 5.0 and 300 mg/kg body veight/day. Moreover, there was some evidence that VCM en hanced the formation of intra-abdominal mesotheliomas and adenocarcinomas of the mammary gland. In females of the low-dose group there was still a high incidence of liver-cell tumours, viz. 28/58. Thus, this study showed a) that VCM is a carcinogen in rats when administered orally, and b) that the "no observed-adverse-effeet level" of VCM in rats with respect to the induction of tumours was lower than 1.7 og/kg body welght/day. For extrapolating experimental data to man In a reliable way the infor mation obtained from such a test system ideally should Include both a "minimum tumour--level" and a "no tumour-level". Therefore, a similar life-span oral carcinogenicity study with VCM in rats was carried out, at lower dose levels, vir. nominally 0.017, 0.17 and 1.7 mg VQi/kg body weight/day (actual oral exposure levels 0,014, 0.13 and 1.3 mg VCM/ kg body welght/day) and two control groups. Based on the pathological findings of the previous long-term study, hlstopathological examinations in the present experiment were focused on the detection of liver lesions, Zymbal gland tumours, mammary gland carcinomas and abdominal meso theliomas. A proposal for the present study - dated 19-02-1979 - was accepted by the sponsor. The present experiment and results are described In this report. S9 9 $ y V 83.285 2. MATERIAL AND METHODS 2.1 Materials Vlnvl chloride monomer (VCM), from Akzo Zout Chemle, Rotterdam, The Netherlands, Physical chemical constants: MW - 62.50; m.p. - -153.8 *C; b.p. - -13.37 *C; density - 0.9106; 1.3700. The product (colourless, clear, free of suspended matter) was delivered in pressurized stainless-steel cylinders, and was specified by the supplier as: Vinyl chloride monomer i. 99.97 wt Z min.; acetylene 2 pl/1 (gas); mono-vlnylacetylene ( 15 jil/1 (gas); 1,3-butandiene (. 10 ^il/1 (gas); methyl chloride 75 jil/1 (gas); ethyl chloride 4 50 jil/1 (gas); chloro- prene i 1 pl/1 (gas); 1,1-dichloroethane 4 1 pl/1 (gas); 1,2-dichloroeth- ane t 20 ^1/1 (gas); acetaldehyde 4.5 mg/kg; hydrochloric acid 4 1 mg/kg; Iron 0.5 mg/kg; water < 100 og/kg; evaporation residue 4 10 mg/kg. PVC--powder. commercial name Carina S 65-02, was supplied by Shell Nederland Chemie, Pernls, The Netherlands, in closed steel barrels. The particle size distribution (by weight), specified by the supplier, was: 0.1 Z max. > 300 fim; A X max. > 200 pm; 90 X max. > 88 pm; 95 Z max. 40 pm. The remaining part of a batch of 500 kg PVC-powder (VCM content 3 ppm) received on January 13, 1977 from Shell, which had been partly used In the previous study (Feron et al., 1978), was used. Part of this PVC-powder was freed from residual VCM by keeping it in layers of 4 to 6 cm thick in a vacuum oven at 60 `C for a period of 3 to 4 days. The so treated PVC-powder (VCM content less chan 0.2 ppm) was used for control purposes. An amount of about 50 kg PVC powder was mixed with a calculated amount of the liquid VCM in a closed steel barrel to raise the VCM content of the PVC-powder to approximately 4600 ppm. This PVC-powder was repacked and stored in tightly closed steel con tainers (containing about 10 kg PVC) in a freezer at -20 *C until u few minutes before mixing with each diet. R&S 134167 R&S 134168 V 83.285 -9- 2.2 Preparation and administration of the diets Preparation and administration of the diets were done In the same way as In the previous life-span study (Feron et al., 1978), Only one distinction has to be made. In that study the dietary levels of VCH were obtained by incorporating into the diets 10 Z FVC-powder with varying proportions of VCM-contalnlng and "VCM-free" powder. In the present study, the diets were prepared to a total concentration of PVC-powder of 1 * only. The proportions of FVC-powder with or without VCM for each of the diets are specified below: group PVC-oouder in Che diet () PVC-powder con taining about PVOpowder without 4600 ppm VCM VCM1) control group low--dose group mid-dose group top-dose group extra control group 0 0.01 0.1 1 0 1 0.99 0.9 0 1 ^ PVC-powder freed from VCM. The VCM-content was lower than 0.2 ppm Diets were prepared daily - by mixing appropriate amounts of PVC-powder (with or without VCM) with the Institute's rat stock diet - Just prior to offering the diets to the rats. The composition of the stock diet is presented in Annex 1. The level of nutrients and contaminants in the stock diet are determined twice a year. The contaminants determined are tabulated, together with det' ction limits and tentative maxima In Annex 2. Actual levels in batches produced on January 24, 1980, September 9, 1980, February 27, 1981, September 9, 1981 and February 9, 1982 are given in Annexes 3 to 7 (nutrients) and in Annexes 8 to 12 (contaminants). R&S 134169 V E3.285 10 As In Che previous study, the pondered diets were available to the rats each day for a period of four hours (generally between 10.00 a.Q. and 02,00 p.a.), la such quantities that the animals did not consume all the feed. At the end of the four-hour feeding period the feeders were removed from the cages and the remainder of the diets was destroyed. The rats had constant access to bottled tap water. The contaminants in the drinking water which are determined twice a year, are given in Annex 13. The actual levels determined In November--December 1979, January--April 1980, September--November 1980, March-Hay 1981, September--November 1981 and January--March 1982 are given in Annexes 14 co 19. 2.3 Animals and housing Five hundred and twenty-five male and 525 female, SPF-bred, weanling rats (Cpb:WU; Wistar random) were obtained from the Central Institute for the Breeding of Laboratory Animals TN0, Zeist, The Netherlands. The animals arrived on August 14, 1979, when they were about 25 days old. The body weight renge Was 38-71 g for males and 42-66 g for females. Upon arrival, the animals were checked for overt signs of ill health and anomalies. One rat showing hydrocephalus, one rat with closed eyes and two rats with a dirty yellow fur were discarded. The healthy animals were allocated to five main groups by a computer randomisation program. Four of the groups, vli. che control group, the low- and mid-dose group and the extra control group, each consisted of 100 males and 100 females. The top--dose group consisted of 50 males and 50 females. To each dietary group a satellite group, comprising 10 males and 10 females each, was attached. The rats of che satellite groups, which received the same diets as the rats of the corresponding main groups, were killed for determinations of glutathione levels in the liver, half of the animals when they had been on their diets for nine months and the remaining half after an experimental period of 18 months. The rats which were not allocated to the main or satellite groups were kept In reserve for a period of four weeks. One female rat of the extra control group showing epileptic fits had to be exchanged for a reserve two days after the etert of the study. None of the other reserved were used. V 83.285 -U The rats were housed under conventional conditions, five rats per sex per cage, in suspended stainless steel cages (17 I ^ x 32 cm), fitted with wire mesh floors and fronts. Animals which were in bad condition were housed Individually in separate cages until they died or until they were killed because their condition was so bad that they were not expected to last out the night, (or, when observed on Friday, the weekend). If the condition of the singly housed rats improved they were returned to their original cage. The male racs of the control-, low-, aid- and top-dose groups were housed in one room, the females of these groups in a second room and the males and females of the extra control group In a third room. The rooms were ventilated with about 10 air changes per hour. The temperature uas kept at 23 + 1 *C, the relative humidity was 40-80 I and a 12 hour light/dark cycle was maintained (light from 06.00 a.m. till 06.00 p.m.). After an acclimatisation period of 7 days, the animals were placed on the test diets for four hours a day during the entire test period. During a period of 3 8ays, prior to the start of the experiment, the rats received stock diet without PVC for four to six hours each day. This was done to adapt the animals to the daily feeding-period of four hours. Drinking water was supplied in glass bottles, which were filled daily with fresh tap water, except for the weekend, and cleaned once weekly. 2.4 Identification of the test system The individual animals were identified in the following way. Each group of rats (the control, low-dose, mid-dose, top-dose and extra control group) uas fitted with a letter and colour code. A table showing the group letter, colour code, exposure level and number of animals la given on the next page. * R&S 134170 i i- v r L CO V 83.285 group colour letter code A white B blue C green D red E brown nominal actual exposure levels exposure levels (mg VCM/kg b w./day) 00 0.017 0.014 0.17 0.13 1.7 1.3 00 number of rats main grouos satellite groups study no. 125 study no. 126 males females males f emales 100 100 100 100 100 100 10 10 10 10 i0 10 50 50 10 10 100 100 10 10 Each rat was identified by a six digit computer reference number, which was even for males and odd for females. Within each subgroup of 10 rats, each rat was Individually identified by sex and one out of 10 different V-shaped earmarks according to the following code: z zero R1L1 " one right plus one left R1 * one right R1L2 " one right plus two left R2 " two right R2L1 two right plus one left L1 one left r2l2 two right plus two left L2 two left R3 - three right See also the cross reference listing (Annexes 20 and 21). Each cage was provided with a coloured card showing the computer refer ence number range, the earmark range, the cage number, the group letter and the study number. 2.5 Experimental conduct The experiment (main and satellite groups) was started on August 21, 1979, Half of the rats of the satellite groups (Assay no. 126) was killed on Hay 20 or 22t 1980 and the remaining half on February 26 or 27, 1981 for determinations of glutathione in the liver. The final autopsy of the rats in the main groups (assay no, 125) was carried out on Juae 24 to 29, 1982, when about 80 Z of the control-, low-dose and aid-dose rsts had died. The following determinations were made: I ! ! | V 63.285 a) Analysis of VCM In the diets 13 - The VCM concent of freshly prepared diets was generally determined once every fortnight. During the course of the study, 72 determinations for each dose level were carried out. Ac the same time the VCM content of the FVC-powder was determined. The rate of evaporation of VCM from the diets was determined by measuring the VCM content of the diets at the beginning of the feeding-period and after 1 hour and after 2, 3 and 4 hours. The diet samples to be analysed were taken at random from the feeders. In this way samples of each of the test diets were taken in week 3, 12, 27, 56, 79, 105 and 132, and were analysed for their VCM content. At the same time samples from the control diets were taken at the beginning of the feeding-period only and also analysed. The analyses were carried out by means of gas-liquid chromatography ac cording to a method described by Feron et al. (1975). b) Analysis of VCM In the faeces Freshly produced faeces from 5 rats of each dietary group were collected 24 hours after the start of the feeding--period at about 10,00 a.m. in week 6, 28, 52, 80 and 106. Fresh faeces was obtained by squeezing the lower part of a rat's abdomen. The droppings were weighed, submerged in 10 ml ethylacetate, and stored at 4 *C in a closed vessel until analysis of the supernatant liquid by gas chromatography. c) Calculation of the actual oral intake and oral exposure levels of VOl R&S 134172 The amount of food eaten by the rats of each test group (g/kg body weight/day) was calculated for males and females separately from the grand averages of body weight (table 7) and food intake (table 9) of each test group over the whole experimental period. V 83.285 14 The VCM intake (mg/kg body ueight/day) vas calculated from the rate of evaporation of VCM from the diets (table 2) and the rate of overall food consumption over the four-hour feeding-period for males and females (table 10). goth the rate of evaporation and the rate of food Intake were assumed to be linear within each hour of the feeding-period. The actual VCM intake for each group during eachof these four periods was calculated by multiplying the amount of food eaten during a one-hour period by the average VCM content of the food eaten in that period. The total actual VCM intake was obtained by adding the VCM intakes during the four one-hour periods of the feeding-period. To obtain the actual oral exposure levels of VCM, the actual VCM intakes were corrected for the VCM excreted with the faeces. For that purpose, the faeces production (g/kg body ueight/day) was calculated basedon an average of 36 g wet faeces/100 g food consumed. The amount of VQ( in the wet faeces (mg/kg body veight/day.) was calculated by multiplying the amount of faeces produced/kg body weight with the VCM content measured in the faeces (table 3). The amount of VCM excreted with the faeces, ex pressed in mg VCM/kg body velght/day, was substracted from the actual oral intake of VCM (also expressed in mg VCM/kg body velght/day) to find the actual oral exposure level of VCM, d) Clinical signs (main groups) R&S 134173 The animals were observed daily and carefully examined for signs of ill ness and tumours once every two weeks. All signs of ill-health or tox icity with any changes in behaviour were recorded, together with the pro gression or regression of such abnormalities, as well as mortality and the time of onset, dimensions and location of palpable tumours. e) Bodv weight (main and satellite groups) The weight of each animal was recorded when the administration of the test substance was started and further in week 2 and 4 and once every four weeks thereafter. From these individual figures the mean body veighi of the animals of each group was calculated. v 83,285 ) Food consumption; amount and rate (main groups) 15 - The food consumption was measured in 20 animals/sex/group, in week 1, 2, 3 and 4, 11 and 12, 23 and 24, 35 and 36, 47 and 48, 59 and 60, 73 and 74, 85 and 86, 97 and 98, 109 and 110 and 121 and 122. The rate of food consumption was determined by weighing the feeders initially and after periods of 1 hour, 2 hours, 3 hours and 4 hours. This was done for each of the diet groups on one day in week 6, 12, 25, 38, 52, 64, 77, 90, 103, 116 and 129. The number of rats involved in these determinations was 20/sex/group. g) Haematology (satellite groups) R&S 134174 Samples of blood were collected from the tip of the tail of 10 rats/sex/ group on days 94 and 185 and of five rats/sex/group on day 367. All blood samples were examined for the following parameters. -- thrombocyte count, by means of the Sysmex Platelet Counter PI 110 using the manual of Toa Medical Electronics Co., Ltd., Japan - prothrombin time using Normo-test reagents from Nyegaard 6 Co. As, Oslo, Norway (Owren, 1959). h) Glutathione in the liver (satellite group) In week 40 and 80, five rats/aex/group were killed by decapitation. The livers were quickly removed and cooled to 0 *C and cut into two, about equal, pieces; both were weighed and stored in a freezer at -20 *C until analysis. After thawing, each sample was homogenised in ice-cold 0.1 M EDTA(Nsj) solution. A part of the homogenates was centrifuged at 2800 rpm for 5 minutes. The non-protein bound sulfhydryl ('glutathione') con tent of the supernatants was determined with 5,5'-dithlobls-(2-nitrobenzoic acid) by means of an AutoAnalyaer. I) Pathology All Bales still alive in week 149 (June 24 and 25, 1982) and all females in week 150 (June 28 and 29, 1982) were killed by decapitation, autopsied and subjected to a careful gross examination. At that tine a mortality of about 80 Z had been reached in the control, low- and mid-dose groups. A thorough autopsy was also performed on rats found dead or killed in ex tremis , All superficial tissues, including the urogenital orifices and tail, each pinna, eye and external auditory meatus, were examined visually and by palpation for distortion, swelling or evidence of tumour formation; simi lar attention was given to the mamnary tracts and subcutaneous struc tures, The external nares, buccal cavity and tongue were then examined, and the cranial roof removed to allow observation of the brain, pituitary gland and cranial nerves. After ventral midline incision and skin reflec tion, all subcutaneous tissues were examined, including regional lymph nodes, mammary and thyroid/parathyroid glands. The condition of the tho racic viscera was noted, and attention was given to the thymus, lymph nodes and heart. The abdominal viscera were examined before-and after removal. The lungs were removed and all pleural surfaces examined. The kidneys were incised and examined. Any abnormalities in the appearance and size of the gonads, adrenals, uterus, intra-abdominal lymph nodes and accessory reproductive organs were recorded. Any lesion suggestive of neoplasia was noted, including details of location, size and multiplicity. Any evidence of adhesion or possible invasion to adjacent structures was noted. Samples of the following tissues and organs of all animals were preserved in 4 I aqueous neutral phosphate-buffered formaldehyde solution: adrenals aorta ovaries pancreas axillary lymph nodes brain caecum cervix coagulating glands parotid salivary glands preputial glands pituitary prostate seminal vesicles <. . *4 r t. r r' r L, u f" L !" t I J '' ^ ' I- L. L V 63.285 17 - colon skeletal muscle duodenum epididymides exorbical lachrymal glands skin spinal cord spleen eves sclacic nerve sternum (with bone marrow) stomach heart submaxillary salivary glands ileum jejunum kidneys sublingual salivary glands testes thymus liver thyroid lungs trachea mamary glands urinary bladder mesenteric lymph nodes uterus nose 2ymbal glands oesophagus all gross lesions Tissues required for microscopic examination were embedded in paraffin wax, sectioned at 5 pm and stained with haematoxylln and eosin. Histopathology was restricted to the liver, all grossly visible tumours or presumable tumours in the abdominal cavity, the glands of Zymbal, and the mammary glands. As to the liver, of each rat three liver pieces, each taken from a different lobe, were studied; these pieces were always taken from the same three lobes at about the same sites; In addition sections were prepared from liver tissue showing gross changes. 2.6 Statistical analysis Data on body weights were subjected to one-way analysis of (co-)variance, followed by Dunnett's multiple comparison test. Thrombocyte counts and prothrombin time values were analysed by the Mann Whitney D-test. Levels of glutathione in the liver were evaluated by the Student t-test. Data on mortality and microscopical observations were analysed by the Fisher exact probability test. Crossly visible masses were evaluated by the Chi square test. L V 83.285 2.7 Contributors f Major contributions to this study were made by: I Animal handling and: Ms. M.M. Andrlnga R&S 134177 diet preparation J.M. Blom Ms. A. Dijkstra Diet analysis Clinical chemistry Ms. G.C.M. Fleer Ms. a.A. van Tuyl D.C. Veldhuysen A. Schouten Dr H.E. Falke J.F. Catsburg Biochemistry Histotechnique Dr D.C. Leegvater Ms. M.T. v. Rijsvijk Ms. J. Wlsman Ms. A.P.M. v. Tuyl-Fonville Ms. N. Hagemeyer Pathology : H.E. Immel Dr V.J. Feron Study director : Dr H.P. Til Study supervisor : Dr V.J. Feron L, " If - 2.8 Deviations from the protocol - At the request of the sponsor, the glutathione determinations in the liver were carried out after 9 and 18 months instead of after 6, 12 and 26 months. - Prothrombin time and thrombocyte count in rats of the satellite groups after 26 months could not be carried out, because all rats of the sat~ elllte group had already been killed before that time. L T~ V 83.285 3. RESULTS 15 3.1 Analysis of VCM In the dices (cables 1 and 2) The average VCM concent of freshly prepared test diets appeared to he 0.49, 4,49 and 44.1 ppm for the low-, mid- and cop-dose group, respect ively (table 1). The VCM content in all three test diets decreased with time during the four-hour feeding-period (table 2). The greatest losses were observed during the first hour. Proportionally, the losses of VCM were virtually the same in all three test diets at the corresponding points of time dur ing the four-hour period. On average, the cumulative losses were 22, 30, 39 and 38 I after 1. 2, 3 and 4 hours, respectively. 3.2 Levels of VCM in the faeces (table 3) The VCM content of the faeces increased with increasing dietary levels of VCM. No appreciable differences in VCM content of the faeces were found within one particular test group. The average amount of VCM found in the faeces expressed as percentage of the actual oral VCM intake, was found to be 23.5, 25.9 and 23,9 I for the low-, mid-, and top-dose groups, re spectively. 3.3 Actual oral exposure levels of VCM (table 4) The VCM concent of the freshly prepared teat diets (table 1) formed the basis for calculating the average VCM content Of the various diets. Since the loss of VCM from the diets during the foul--hour feeding-period (table 2), the rate of food Intake (table 10) and the VCM content of the faeces (table 3) were knoun, the actual oral exposure levels of VCM for each group could be calculated (see section 2.5). They were found to be 0.014, 0.13 and 1.3 mg VCM/kg body weight/day for the low-, mid--, and top-dose groups, respectively (table 4). V 83.285 2C - When the actual oral intake of VCM uas expressed as percentage of the theoretical intake, the values found were 81.5, 81.2 and 81.3 X for males, and 80.1, 79.8 and 79.8 X for females of the low-, mid- and topdose groups, respectively. The overall average for males and females of the various test groups was calculated to be 80.6 X. This percentage uas in good agreement with the 80 X found in the previous life-span study using much higher dietary levels of VCM (Feron et al. , 1978; 1981). 3.4 Symptomatology and survival (tables 5 and 6) There were no overt signs of reaction to treatment with VO.. The behav iour of the rats during the first 18 months of the experiment was unre markable. After 18 months ageing symptoms developed in all groups and the number of unthrifty rats increased, more rapidly in males than in fe males. The poor condition started with a humpbacked position and slight emaciation, followed by dyspnoea, pale eyes, lethargy, filthiness and in many cases severe emaciation. Randomly distributed major abnormalities due to ageing included maloc clusion of incisors, staired coats, a bloody discharge around nostrils and eyes, wet stools, focal alopecia, focal dermatitis, paresis of hind legs, loss of one or both eyes and white opaque cornea. Macroscoplcally visible or palpable masses occurred in all groups. The total number of rats which showed such a lesion at any time, together with the number of masses is given in table 5. In females both the inci dence and the total number of these gross lesions was considerably higher than in males. Taking into account that the top-dose group comprised only 50 rats of each sex, the total number of masses in females of the topdose group uas higher than in the other groups. This remarkable differ ence appeared to be caused by Che high number of top--dose females with liver cysts (see also tables 13 and 14), which are easily detected by palpation of the abdomen. In the extra control group the total number and the incidence of masses was relatively low as compared to the other groups, the control group included. The low figures for the extra control group are undoubtedly related to the relatively high and early mortality in this group, giving, fewer animals of this extra control group the time to develop lesions associated with old age. v 83,285 21 - The cumulative mortality Is given In table 6. Up to week 66 mortality was very low. Thereafter, mortality gradually Increased, more quickly In che extra control group than in the other groups. The extra control animals which were housed in a separate room - were much more severely affected by chronic respiratory disease than the animals in the other groups (see also 3.9.1). This phenomenon undoubtedly explains che higher and earlier mortality in the extra control group. 3.5 Body weights (tables 7 and 8; figures 1 and 2) In the three test groups, the mean body weights were generally comparable to those of the controls. Some isolated changes were observed, but there was no evidence of a dose-related response. In the extra control group mean body weights were statistically signifi cantly decreased throughout the study in males and from day 674 and on wards in females as compared to the standard control group. In males the lower body weights were accompanied by lower intake of food (see 3,6). Since the extra controls were housed in a separate room, the lower body weights and food consumption might be due to unknown differences in en vironmental condition (see also 3.4), although this separate room was conditioned in the same way as the other rooms. 3.6 Food Intake (tables 9 and 10) Food intake showed wide variations among the different weighings within one group. There was, however, no evidence that food Intake was affected by VCM-feeding (table 9). Hales of the extra control group had a lower food Intake than the other groups from week 3 onward. No appreciable variations were encountered in the rate of food consump tion in the various groups (table 10). Females consumed their food slightly less quickly than males. R&S 134180 R&S 134181 V 83.265 - 23 - 3.7 Haematology (table 11) Thrombocyte count did not shoo dose-related differences between the test groups and the controls. The slight, though statistically significant increase in females of the top--dose group on day 366 was not apparent when compared to the extra controls. Prothrombin time of females of the top-dose group was slightly increased after 3 and 6 months. However, prothrombin time of females of the extra control group was even higher at the same stages. No differences between the groups occurred after a feeding-period of 12 months. 3.8 Glutathione levels of the liver (table 12) There were no significant differences in liver glutathione levels between the various test groups and the controls, either after 9 or 18 months. 3.9 Pathology 3.9.1 Gross examination (table 13) The incidence of liver nodules suspected of being tumours was higher in males and females of the top--dose groups than in controls or lower dose animals. Most of these tumourous masses were small (diameter up to 1 cm), solid and pale or had the same colour as the adjacent liver tissue; some were large (diameter 2 to A cm), soft or firm and occasionally slightly haemorrhagic. Cysts in the liver of old rats, particularly in females, is a common finding in the strain of rats used. However, the incidence of such cysts was much higher in females of the top-dose group than in females of the other groups, the control group included. The cysts varied widely in sire, were often multiple and generally contained a turbid, watery liquid. There was no evidence that any of the other gross lesions observed and summarised in table 13 were related to the administration of VCM. v 63.235 13 - Severe chronic respiratory disease (CRD) often accompanied by focal emphysema (spongy lungs) was seen much more frequently in the extra control group than in the other groups. Many of the rats affected by ad vanced CRD lost weight and emaciated before they died or were killed in extremis. Other conspicuous findings in animals of the extra control group were the high incidences of bilateral testicular atrophy and atrophy of the coagulating glands and seminal vesicles. Atrophy of the testes and of the secondary sexual glands were often seen in the same animals, but these lesions also occurred separately. Moreover, a relatively high number of females of this group showed protrusion of the eye balls; this was almost exclusively observed in severely emaciated rats. 3.9.2 Microscopic examination Liver (table 1A) Increased incidences of foci of cellular alteration, Deoplastlc nodules, hepatocellular carcinomas, livei--cell polymorphism and cysts have been observed in the top-dose group. Moreover, in the top-dose group two females and one male had developed a hepatic angiosarcoma, whereas such tumours have not been seen in any of the other groups. Since the morphology of the hepatocellular lesions was essentially the same as that of the liver lesions described in full detail in our previous report on Che long-term effects of oral administration of VCM in rats (Feron et al,, 1981), no further dlscription of these alterations is presented in this report. The morphology of VCM-induced hepatic angiosarcomas has also been described in a previous report (Spit et al., 1981). Table 1A also shows that in females, but not in males, of both the lowend mid-dose group the incidence of basophilic foci of cellular alter ation was significantly higher than in controls. In addition, in the mid-dose group the number of females bearing foci of cellular alteration was statistically significantly higher chan in the control group. V 3,285 21 There was no evidence that any of the ocher hepatic lesions observed, was related to the feeding of VCM, There were a few statistically significant differences between test groups and the control group, but this concerned decreases In incidence In the test groups (e.g. slight cholangloflbrosis in males), or the differences were not statistically significant as com* pared to the extra controls (e.g. single cell necrosis in females), or there was no dose-response relationship (e.g. foci of degenerated heparocytes in males). As compared to controls lower incidences of several hepatic abnormalities were found in the extra control group. Examples of such lesions are cysts, bile duct proliferation, cholanglofibrosls and vacuolisation of periportal hepatocytes In females, and liver cell polymorphism In males. Such changes might be associated with old age and their relatively low incidences in the extra control group is most probably due to the much lower average survival time of these animals (118 weeks) in comparison with that of the controls (129 weeks). R&S 134183 Mammary glands (table 15) The number of females bearing a qammary gland tumour was considerably lower in the extra control group and in the low- and mid-dose groups than in the control group. In the same groups the number of females with a mammary fibroadenoma was also lower than in the control group. There was no indication that the incidence of mammary gland carcinomas Increased with Increasing VCM dose. Abdominal tumours (table 15) u All intra-abdominal tumours found at autopsy were examined histologi r cally. The numbers of mesotheliomas detected in rats of the different L groups indicate that there was no evidence of a relationship between the administration of VCM and the development of this type of tumour. Zymbal gland tumours All gross tumours in the head region were studied by light microscopy. No f Zymbal gland tumour was found. L f. I L L. V 83.285 4. DISCUSSION In the present study no effects attributable to VQ1 vere observed other than liver changes, and a higher mortality in the top-dose group as com pared to the control group during the last 6 to 9 months of the exper imental period. The differences In mortality between the top-dose group and the control group were slight and attained a level of statistical significance only at a few points of time. This slightly increased mortality in the topdose group might be due to the Increased Incidence of neoplastic and non-neoplastlc liver changes in this group. A similar adverse effect has been observed in females, but not in males, of the low--dose group (1.7 mg VQi/kg body veight/day) used in the previous long-term oral rat study with VCM (Feron et al., 1981). From the previous study (Feron et al., 1981) it appeared that oral exposure to VCM at a level of 1.7 mg/kg body weight/day resulted in a series of changes in the hepatic parenchyma. These changes Included an Increased Incidence of cellular alteration and neoplastic nodules, a few hepatocellular carcinomas, an increased Incidence and degree of liver-cell polymorphism, and an Increased Incidence of hepatic cysts. In the present study the same type of treatment--related liver changes were found in the top-dose group receiving 1,3 mg VCM/kg body weight/day. In addition, one male and two females of the top--dose group had developed a hepatic angiosarcoma; in the previous experiment hepatic angiosarcomas were observed at VCM levels of 5.0 mg/kg body weight/day and above but not at the lowest exposure level of 1.7 mg/kg body weight/day. As compared to controls Increased incidences of basophilic foci of hepatocellular alteration occurred in males of the top-dose group and in females of each of the dose groups. In females the Incidence of this type of foci of cellular alteration in the liver increased with increasing dose levels. Moreover, the number of rats bearing foci of cellular alteration in the liver (all type of foci) was significantly higher in males of the top--dose group and in females of the mid-- and top-dose group than in controls. These findings suggest a relationship between the oral exposure to VCM at levels of 0.014 and 0.13 mg/kg body weight/day and the Increased occurrence of foci of cellular alteration lu the liver of female rats. V 83.285 26 The incidence of basophilic foci of hepatocellular alterations in females of the control group (9/98 or 9 I) was clearly lower than the Incidence of this type of foci in female controls used in the previous long-term oral rat study with VCM (10/57 or 18 I; Feron et al., 1981). Similar Incidences were also found in females of control groups of three other long-term rat studies carried out in our Institute more or less simultaneously with the study of Feron et al. (1981); the Incidences being 11/95 or II I, 3/20 or 15 I and 17/99 or 17 Z. These data seem to indicate that the incidence of basophilic foci (9 I) in females of the control group of the present VCM study is unusually low, and, thus, would seem to throw doubt upon the toxicological significance of the slight increase in incidence of this type of foci of cellular alterations in the low- and mid-dose groups. However, the incidences of basophilic foci of hepatocellular alterations in females of control groups of other long-term rat studies performed in our Institute in the same period as the present VCM-study were: 0/20 or 0 ;, 1/20 or 5 I, 5/69 or 7 I and 1/50 or 8 Z, Compared to these figures an incidence of 9/98 or 9 I as found In the present group of female controls Is not unusually low, and, thus support the suggestion that the feeding of VCM at levels of 0.011 or 0.13 mg/kg body welght/day may have resulted in more basophilic foci of cellular alteration in the liver of female rate. The directive of the European Community (EC) of January 30, 1978 (7S/112/EEC) Imposes a maximum permitted limit of 1 ppm residual VCM in finished plastic materials or articles Intended for use in contact with foodstuffs. The same EC-direetive stipulates that any migration of VCM to the food must not produce a concentration in the food of greater than 10 ppb VCM which is or is very close to the detection limit of VCM in foodstuffs. Calculations based on a survey of residual levels of VCM in PVC bottles, films and foods in the period of 1974-1977, suggest that the intake of VCM from the average diet will be less than 0.1 pg per person per day (Crosby, 1982; Ministry of Agriculture, Fisheries and Food, 1978). What does this maximum oral daily Intake of VCM by humans mean in terms of cancer risk? R&S 134185 R&S 134186 V 83.285 - 21 - Extrapolation of the results of the present rat study to humans using the linear extrapolation model and taking into account en acceptable cancer risk from oral VCM exposure for man of 10 , leads to an acceptable oral intake of 0.4 pg VCM per person per day. Since the estimated maximum oral dally Intake is only 0.1 pg VCM/person/day, the estimated cancer risk is at least a factor 4 lower. Moreover, the extrapolation model used is a conservative and prudent one, which implies that the actual cancer risk night be (considerably) lower than that calculated by means of the linear model. In addition, comparison of the results of an epidemi ological study of vinyl chloride workers with the predicted cancer risk in the cohort of workers based on the results of VCM studies in rats suggested that humans are less sensitive to the carcinogenic action of VCM than rats (Gehrlng ec al., 1979). Translated into practical terms this means that the cancer risk of a maximum oral dally Intake of 0.1 pg VCM/person/day is small enough to be practically neglected. 5. CONCLUSIONS The results of the present oral study allow the following conclusions: - VCM at a level of 1.3 mg/kg body welght/day induces neoplastic and non-neoplastic changes in the liver of male and female rats, - VCM ac a level of 0.13 mg/kg body weight/day may lead to more female rats bearing foci of cellular alteration in the liver, - VCM at levels of 0.014 or 0.13 mg/kg body weight/day may result in an increased incidence of basophilic foci of cellular alteration in the liver of female, but not of male, rats, and - 0.13 mg VCM/kg body weight/day is a 'no-observed-adverse-effect-level with respect to the induction of tumours in rats. R&S 134187 V 83.2S5 6. references Ministry of Agriculture, Fisheries and Food (1978). The Surveillance of Food Contamination in the United Kingdom: Survey of Vinyl Chloride Content of Polyvinyl Chloride for Food Contact and of Foods. The Second Report of the Steering Croup on Food Surveillance - The Working Party on Vinyl Chloride. Food Surveillance Paper No. 2, p. 6. HMSO. London. Crosby, N.T. Analysis for residual monomer levels In plastic and in foods Anal. Proc. September 1982, 426-430 Feron, V.J., et al. Life-span oral toxicity study of vinyl chloride in rats CIVO-Report No. R 5788 (1978) Feron, V.J,, et al. Lifespan oral toxicity of vinyl chloride in rats Fd. Coamet. Toxicol. 19 (1981) 317-333 Feron, V.J., et al. Observations on the oral administration and toxicity of vinyl chloride in rats Fd. Cosmet. Toxicol. 13 (1975) 633-638 t Cerlng, P.J., P.C, Watanabe and C.N, Park ( Risk of angiosarcoma in workers exposed to vinyl chloride as predicted from studies in rats Toxicol. App. Pharmacol. 49 (1979) 15-21 Ovren, P.A. Thrombotest; a neu method for controlling anticoagulant therapy Lancet, 1959, II, p. 754 L Spit, B.J., V.J. Feron and C.F.M. Hendrlksen Ultrascructure of hepatic angiosarcoma in rats Induced by vinyl chloride Exp. Mol. Pathol. 35 (1981) 277-284 L L f I V 83.285 29 Squire, R. A. and M.H. Levitt l Report of a workshop on classification of specific hepatocellular lesions in rats Cancer Res. 35 (1975) 3214-3223 r L r m r r [ c L f L. f- L INST. CIVQ-TOX. AND NUTR. TNO 83-09-01/AE r i f I 30 03 CJ 4* 03 00 V 83.285 7. AUTHENTICATION This report was prepared by: date: I (Study director) \ t r i L r l L r L. 3J fto CO CO ft. --t 09 (O r Lt (Pathologist) (Study supervisor, Pathologist) and approved by: Dr A.P. de Croot date: (Head Dept. Biological Toxicology) S/L^dc------/f 8 5 R&S 134190 V V 83* 285 - ) ' b. RETENTION OF RECORDS, SAMPLES AND SPECIMENS " 31 - j~ All records concerning the dietary formula, daily observations, body [ weights, food Intake, haematology, clinical chemistry and gross- and microscopical examinations and all other information relevant to the p- quality and integrity of the study have been filed in the archives of the j. Department of Biological Toxicology together with the master copy of this report. Materials (samples of the test substance, wet specimens, blocks) r. have been stored, and will be retained for a period of eight years, i.e. [ till July 1990. The slides will be retained for a period of IS years, i.e. till July 1997. r f' L L R&S 134191 V 63.2S5 3: QUALITY ASSURANCE UNIT TNO - I'.O. Box 360, 3700 AJ ZEIST, Netherlands STATEMENT OF CLP COMPLIANCE On : Lifespan oral carcinogenicity study of vinyl chloride in rats Report no.: v 83.285/291099 Date : September, 19S3 The study was carried out under conditions of good laboratory practice. Within reason there have been no circumstances that might have affected the quality and integrity of the results obtained. Dates and number of inspections: 19 November 1979 22 and 25 February 1980 (1) (1) 12 May 1980 12 November 1980 17 November 1980 23 January 1981 3-4 March 1981 14-15 May 1981 (1) (1) (2) (1) (2) O) 18 August 1981 27 October 1981 (1) (3) 22 January 1982 (3) 12-13 May 1982 (2) 24 June 1982 8 July 1982 CD (3) 8-11 March 1983 (2) 10 March 1983 (1) Draft report audit: 10-11 March 1983 Dates of reports to management: 19 November 1979 25 February 1980 12 May 1980 12 November 1980 18 November 1980 26 January 1981 5 March 1931 18 May 1981 18 August 1981 27 October 1981 26 January 1982 13 May 1982 24 June 1982 8 July 1982 16 March 1983 24 March 1983 24 March 1983 Final report audit: 20 September 1983 (1) 20 September 1983 Drs S. van Straten Quality Assurance Manager date: V 83.285 CIVO/TNO STUDY NO 125 CARCINOGENICITY STUDY IN RATS WITH VCM IN THE DIET TABLE 1 LEVELS OF VCM IN THE DIET IMMEDIATELY AFTER PREPARATION OF THE DIETS ' Croup A B C D E Intended oral exposure Dietary level of VCM (ppm) levels of VCM MEAN SEM N (og/kg b.v./day)_______ 0 < 0.1 72 0.017 0.487 0.013 72 0.17 4.487 0,097 72 1.7 44.07 0.926 72 0 < 0.1 65 1) The VCM content of the PVC~powder with VCM was on average 4582 + 98 ppm. R&S 134192 f E6tfrSl. SSb -- -I 1 C1VO/TNO STUDY NO 125 CARCINOGENICITY STUDY IN RATS WITH VCH IN THE DIET TABLE 2 LOSSES OF VCH FROM THE DIET DURING A TERtOD OF FOUR HOURS Croup Intended oral exposUlre levels of VCH (tng/kg b.u,./day) VCH lcve1s In the 'diet during the four-hour feed Ing"pcrlod at different points of time ^ expressed. as ppm 2) expressed as percentage of to fo *1 1 2 c 3 b, ll 12 1 3 l4 A 0 < 0.1 < 0.1 ' 0.1 < 0.1 * 0.1 B 0.017 0. A 8 0.37 0.32 0.27 0.28 77 67 56 58 0.0373) 0.039 0.024 0.021 0.016 C 0.17 A.2 3.3 3.0 2.7 2.7 79 71 64 0.36 0.23 0.13 0.20 0.15 D 1.7 41.3 33.3 29.9 26.7 25.3 81 72 65 61 1.96 1.85 1.70 1.66 1.96 E Grand Mean 0 < 0.1 < 0.1 < 0.1 < 0.1 < 0.1 79 70 62 62 ^ Samples taken Immediately after preparation of the diets C t q J and further from the feeders In the crtp.cn of the rnts one hour (t^), two hours (tjK three hours (t^) ami four hours (t ^) after preparation ^ The values are the means of 1 different analyses. ^ Standard error of the me1'. i f i r--, r ~1 -~r) --! '*! CIVO/THO STUDY NO 125 CARCINOCENICITY STUDY IN RATS WITH VCH IN THE DIET TABLE 3 LEVELS OF VCH 111 THE FAECES Croup A B C D E Intended oral exposure levels of VCH (tnft/kR b.v./day) 0 0.017 0.17 1.7 0 Level of VCH (ppm) found In the faeces 2 4 hours after Lhe start of the fcedlnfc-period* ^ MEAN SEH N A o H- 5 0.26 0.028 5 2.64 0.273 5 23.6 2.968 5 < 0.1 5 1) Faeces were collected by'squeezing tbe lower part of the ra t ' b abdomen; they were Immediately submerged In 10 ml ethylacetate and then stored at 4 'C In a closed vessel until analysis for the VCH content. i i ) .' t- w mm ssy nr i T' r c~ r~--5 r " - -- -----i -- ; --3 ---i --i 96 l- S^H CiVO/TNO STUDY NO 125 CAKCINOGEN1C1TY STUDY IN RATS WITH VCH IN THE. DIET TABLE A DESIGN AND ACTUAL DOSE LEVELS OF VCH IN RATS FED DIETS CONTAINING I'VC POWDER Group A B C D E Intended oral exposure levels of VCH ppm mg/kg b.v./ In diet 0 /day 0 0.34 0.017 3.4 0.17 34 1.7 00 Actual dietary VCH levels at the start of the feeding period * ^ (ppm) 0 0.49 4.5 44.1 0 Theoretical oral Intake of VQI2) {mg/kg b.v./ day) 0 0.022 0.21 2.1 0 Actual oral Intake of VCH33 (rag/kg h.v./ day) 0 0.018 0.17 1.7 0 Ac t ual oral ex posure level of 4)----VCH fmg/kg 1>. w. /day ) 0 0.014 0.13 1 .3 0 ^.Average dietary VCH contents determined Immediately after preparation of the diets. 2)* Assuming no loss of VCH by evaporation from the diets (see also table 1). ^ Taking Into account the loss of VCH from the diets. A) ' Oral Intake of VCH diminished by the faecal VCH. The VCH excreted In the faeces was considered to be still enclosed In the PVC granules, and thus not to have been In contact with the body. Cl VO/TWO STUDY NO 125 CARCINOGENICITY STUDY IN RATS WITH VCT! IN THE DIET TABLE 5 CROSSLY VISIBLE OR PALPABLE SUPERFICIAL OR INTRA-ABDOMINAL MASSES OBSERVED IN THE COURSE OF THE STUDY C roup Intended exposure levels of VCM (mg/kg b.u./day) Total no. of rats at start No. of rats with masses Tota 1 Hon-survivo rs HALES A0 B 0.017 c 0.17 D 1.7 E0 100 35 100 A 5 100 36 50 18 100 ** 32 AO 31 17 H** No . of masses Total H1sa ppeared In course of time 55 10 55 10 A2 7 23 6 16 I FEMALES A0 100 68 59 B 0.017 100 80 69 C 0.17 100 63 A9 D 1.7 50 38 36 E0 100 48** A8 112 3 119 8 115 6 87 2 79 4 STATISTICS: CHI-sqoate test: ** P < O.Ol, *** P < 0.001 96VVCV SW I r r---?. r zei.m ssa r~- r--1 r-'" i _ . *' . w ** *'"' T ftpL E f> M ALz s DAY A [tftY 28 ti fir Si [if>r 84 : ft r 11 2 [ift r 110 lift Y 148 [ift Y [<n r 194 PAY PAY 252 TO A PAY 308 PAY 334 PAY 34-! PAY 392 Dft Y PAY PAY 120 110 *74 PAY 504 PAY PAY 540 PAY 580 PAY PAY 616 611 PAY 472 PAY 7AA i PAY TOR i PAY 756 MORTftL I TY ............................... ...................... COUTKOI INCIPEMCES o o:: A OV a or; i i:: i 12 i i:: i i:; i 12 i 12 i 12 i 12 i 12 12 a "1 V a 2v A 22 3 32 4 42 1 42 4 42 4 42 4 42 4 62 4 42 4 42 11 11X 12 122 1 3 1 22 0.017 MC-.'KG I HE 1 PE lit E 8 0 02 A 02 0 02 1 12 1 12 1 12 1 12 1 12 1 12 1 12 1 12 1 12 1 12 1 12 1 12 1 12 1 12 1 12 1 42 4 42 1 4 2 "? -*x 8 87 8 ,82 ! 2 122 15 152 1 5 152 15 152 .......... .... , ............... ____ ... - _------------------ . - . _ ................ . . .......................... _ ... 0.17 HG.'KG JHCIPEUCES 0 02 A AV A 02 0 02 0 02 A 02 A 02 A 02 0 02 o AV A 02 l 12 l 12 l 12 A 22 A AV 3 32 3 3 y. 4 42 5 52 4 62 A ? y. "* A 7 7 77 8 82 Cl x 1 3 1 32 1 4 142 1 . 7 HG/KG me I PEHCES 0 02 A r. v r. o y. A AV 0 02 0 02 0 02 0 02 A 02 A 02 A 02 0 02 0 or; 0 02 0 02 o 02 A 02 0 02 1 22 1 Av A 6 7: A /, V A uV *J f V 7 f>;-: 4 82 If. 2 10 202 fnM M [.' L i o !. 1 fL l A A7 r r. 0 A V a ' 0 7 A a; A 0` 0 A v' .A A* A O' A A* A 1'* C' CJ ' 8 E" o C 1 1: ' *' -`O'* L1 '* ir 1 statistics: FISHER EXACT PROBABILITY TEST p-io.os p '0.01 F' 0,001 TIJO SI PEP { r" i r r" TAPLE 6 HA L ES DAT DAT DAY DAY n ay DAY DAY DAY DAY DAY 'BA B12 S 10 B6B S? r 'i g PO 100B 1036 MORTALITY (CONTINUED I) CONTROL INCIDENCES 15 I s:: 31 21" 28 2BX 31 3ti: 3B 30X 14 a a:* 53' 53:: 5? 5?:: 72 nv 80 BOX 0.017 HG/KG INCIDENCES 1 7 1 ->7. it; 25:: 28 2 8:; 34 3 4 X A3 4 3X 51 5 IX 5-1 5 4 X 63 6 3X 69 69X BO BOX 0.17 HG/KG INCIDENCES 16 16X 23 T 1 V 2B 2B7. 34 3 4 X 3 B 3 OX 4 4 4 1X 53 53X 59 59X 72 72X 82 B2X statistics: FISHER EXACT PROBABILITY TEST . P-.0.05 '* P co.oi 1 . 7 HG/KG INC I DEUCES 1 5 H) X 18 If, X 21 i 2 X 24 48 X 24 48 X rjf> y. Z2'* UU7. 35 * 70 X 39 * 78 X 42 ' * * 8A V. PYO.OOl 1UU SIDED CONTROL INC 1 BENIN ' \ ' . * 7Q . . 71 * * > 'A i > '[' 100 . 1 1 00 ' ' I 00 * * * IOC' ~1 : C" 1 C-v 100 100 100 -- I ) 86Vt?V S'SH r 66 SSH TABLE 6 MORTALITY (CO NTINUEP 2) FEMALES PAY PAY PAY PAY PAY PAY PAY PAY PAY PAY PAY PAY PAY PAY PAY PAY PAY PAY PAY PAY PAY PAY PAY PAY PAY PAY PAY PAY 30 50 06 11 4 h: 170 190 226 351 202 310 338 366 394 A 150 478 506 534 563 590 618 646 674 703 730 758 CONTROL INCIPENCES 0 02 0 07. 0 02 0 02 0 02 0 02 0 02 0 02 0 02 0 02 0 02 1 12 1 12 1 12 1 12 1 12 1 12 1 12 2 32 2 32 4 42 5 52 6 62 7 72 10 122 13 122 15 152 20 202 0.017 HG/KG INCIPENCES 0 02 0 02 0 02 0 02 0 02 0 02 0 02 0 02 0 02 0 02 0 02 0 02 0 02 0 02 0 02 0 02 0 02 1 12 1 22 n 22 4 42 4 42 4 42 5 52 8 82 8 82 11 112 13 132 0.17 MG/KG INCIPENCES A 02 0 02 0 02 0 02 0 02 0 02 0 02 0 02 0 02 0 02 0 02 0 02 0 02 0 02 1 12 1 12 2 32 A 32 3 32 3 32 3 32 4 42 4 12 7 72 10 102 13 132 1 4 1 42 19 192 o o statistics: r I SHER EXACT PROBABILITY TEST * P<0.05 * ' P I 1. 7 HG/KG INCIPENCES 0 02 O 02 0 02 0 02 0 02 0 02 0 02 0 02 0 02 0 02 0 02 0 02 0 02 O 02 0 02 0 02 1 22 1 22 2 42 n 4;; 42 5 102 6 122 6 122 6 122 6 122 13 242 1 4 28 2 P',0.001 TU0 SI PEP CONTROL 1 NC1 PENCES 0 02 0 02 r. r, v o 02 0 02 0 A '1 0 f. 0 02 0 02 0 02 0 AV 0 V *, 0 02 0 0'/. 0 02 0 02 0 02 A 02 0 0T 0 O'. 0 02 1 12 n '> * AV 2 AV 7 * IV y/ 1 4 1 12 1 r--: r r~"! r~ i~ i ' i ) "1 ir TABLE 6 MORTALITY (CONTIHUCO 3) F E HALE S bay PAY PAY hay Ii ft Y BAY PAY PAY PAY PAY 7B6 81 A 842 870 B98 ??6 954 9B2 1010 103 B CONTROL INCI PENCES 20 203 26 263 2B 2 B 3 30 303 35 353 41 413 A 7 473 5B 583 66 663 76 763 0.017 hG/KG I MCI PENCES 1 5 153 2 2 53 32 323 36 363 4 1 413 5 1 513 60 603 66 663 72 723 77 773 0.17 hG/KG 1MCI PENCES 1 9 193 nji 27 273 30 303 35 353 45 453 52 523 61 613 67 673 74 743 o o statistics: FISHER EXACT PROBABILITY TEST * F'10,05 1 P 1 .7 HG/KG INC!PENCES 15 10 3 1 6 12 3 1 9 18 3 21 4 2 3 2 A 48 3 28 563 32 1 64 3 36 43 1 * 45 723 863 SO 3 P/0,001 T K0 SI PEP CONTROL INC I PENCES 16 16 3 25 n rj 3 ' Z ? *; 50 1 50 3 58' ' 9 -I * * 5U3 V2*: 100 1003 100" 1003 100 100 3 100` 1003 I I osm sva iozpzi s'sa TABLE 7 HEAH POPY WEIGHTS (G) HALES CONTROL MEAN SEH N 0.017 HG/KG MEAN SEH N PAY PAY DAY PAY PAY PAY PAY PAY PAY DAY PAY PAY PAY DAY PAY PAY PAY PAY PAY JAY JAY JAY 1AY .'AY JAY JAY 1A Y 1A Y 1AY 0 14 28 56 04 112 1 40 I6B 194 224 252 280 308 334 344 392 420 4 48 474 504 532 560 588 616 644 672 700 728 756 53.9 95.7 149.8 233.4 286.4 326.5 344.7 367.5 376.6 386.5 300.5 403.4 411.2 416.5 413.3 410.0 404.2 404,2 405.9 400.4 4 15.6 419.4 414,5 419.4 428.8 431.1 427.0 425.4 419.0 0.5 0.8 1. 4 2.4 3.1 3.4 3.5 3.7 3.7 3.9 4.0 4.0 4.2 4.2 4.2 4.2 4.1 4.2 4.4 4.2 4.1 4.1 4. 1 4.2 4.3 4.2 4.0 4.0 4.2 100 100 100 100 99 99 99 99 99 99 99 99 99 99 98 98 9B 97 96 96 96 96 94 94 94 94 B9 BB B7 53.2 96.9 1 147 . B 223.0 ' 1 271 .0 1 309,2'' 340.2 357.0 370 .B 377.7 301.2 400.0 403.3 406.5 409. I 403.6 397.3 399.0 403.3 395.2 407.5 414.9 4 12.5 4 15.9 425.9 427.5 424.8 421.2 418.5 0.5 1 .0 1.6 2.4 3.0 3.3 3.6 3.7 3.9 4.0 4.0 4 .1 4.2 4.2 4. 3 4.3 4.3 4.4 4.4 4.3 4.4 4.3 4.5 4.5 4*5 4.7 4.8 4.8 4.7 1 00 100 1 00 1 00 9? 99 99 99 99 99 99 99 99 99 99 99 99 99 99 96 96 96 93 92 92 B? B6 95 85 0.17 HG/KG HEAN SEH N 53.2 95.9 148.9 222 * 0 283.3 323.8 343.0 364.7 372.8 385.7 307.0 405.2 407.7 4 15.0 4 11.7 407 . 4 402.9 404.7 410.9 404.0 416.1 422.6 418.4 425.2 431.5 437.8 431.4 428.0 423.3 0.5 0.8 1 .3 2.5 3.0 3. 1 3. 4 3.4 3.6 3.7 3.4 3.6 3.7 3.8 3.9 3.9 3.9 4. 1 4.1 4 .4 4.2 4.2 4.3 4.3 4.4 4.5 4. 4 4. 4 4.6 I 00 100 100 100 1 00 100 100 100 100 100 100 100 99 99 99 90 98 97 97 96 95 94 93 93 93 92 91 87 86 ITATISTICS 1 COVAR f PUNNETT TESTS * p 10 . 05 p:o.,01 TUG SI PEP 1 .7 HG/KG HEAN SEH N 54.3 98.2 149.2 224.5' 279.0 316.6 340.0 364 . 1 304 . 3 309.3 387.3 408.3 414.6 4 17.9 410.6 419.4 411.9 413.1 414.0 409,2 420.6 426.7 423.5 429.3 438,5 4 4 6.5 440.4 428 . B 423.9 0.6 1.1 1 .B 2.9 3.6 3.9 4.4 4,7 4.8 4.9 4. 7 5.1 5,2 5.0 5.1 4.9 4.0 4.B 5.1 5.0 5.0 5.5 5.3 5.5 5.9 6. 1 5. B 7. 4 6.9 50 50 50 50 50 50 50 50 50 50 50 50 SO 50 50 50 50 50 50 49 49 49 4B 4B 47 47 46 44 40 CONTROL HEAN SEH H 51.5 89.3 ' ' 141.7' 214.6 ' ` 251.1" 27B . 1 ' ' 297.5* * 314.5" 31 B . 5 * ' 339 . B ' ' 351.4" 365.0'' 371.0" 371.3" 377.8" 3B5.2" 370.9'' 36B.O'' 377.3'' 364.0" 390.7'' 3B4.3 ' ` 379.9 ' ' 376 . / ' ' 379.7' ' 381 .2 " 365.9' * 357.2 " 344 . B ' ' 0.5 0.7 1.1 1 .5 2,0 *> . 2.3 2.6 2.8 2,6 ' ( -7 2.7 2. B 3.3 J .0 3. 1 2.9 2.9 3.0 3. 7 3.3 3. 3 3.3 3. 7 1 . 1 4.3 4*5 4.9 iOO IOO 100 SOO 100 100 100 100 100 100 100 100 IOO 100 100 100 100 100 9? 92 92 92 92 92 90 07 02 7II 71 (EXP. UNIT = Al 1 HA 1 ) c r- r~ r 1------ 1 r - --------- 1 TABLE 7 HALES MEAN BOOT UEIGKTS (0) (CONTINUED 1) CONTROL MEAN SEh N 0.017 MEAN HG/KG SEH N 0.17 HG/KG HEAN SEH N BAT 784 BAT 812 BAT B 4 0 BAT 868 BAT 096 BAT 924 BAT 952 BAT 900 BAT100B PAT 1036 BAT 1043 408.6 394.0 393.3 387.4 376.6 365.6 361 .7 352.9 346.5 341.3 343.0 4.0 3.9 3.9 3.8 4.3 4.3 4.1 4. 4 5,2 5.2 5.3 05 79 72 69 62 56 47 41 30 21 20 406.3 395.5 400.0 390.6 383.7 372.9 368.7 359.5 360.7 356.6 359.3 4 . B 83 409.4 4.9 75 402.9 4.7 72 400.6 5.0 67 394.8 E . 6 59 386.3 5 . B 49 371 .2 6.4 47 363.4 6 . 1 3B 360.2 6.5 31 35B. 6 5.5 20 356.3 5.6 20 358.1 4.6 4.5 4.7 4.7 4.5 4.4 4.7 4.9 6.3 6.2 6.3 B4 77 72 66 62 56 47 43 20 19 10 1.7 HG/KG HEAN SEH N 414.9 403.5 398.3 393.7 382.5 300.0 360.6 363.3 354.2 366.7 368.7 6.7 7. 1 5.9 5.7 6.1 7.0 7.7 B,0 7.4 7.B 7.9 35 32 29 26 26 21 10 16 11 8 8 CONTROL HEAN SEH 322.9 1 302.5' ' 297.9 ' 299.3' 1 2B0.1 ' 1 329.8 -- --- -- 5.4 5.5 7.0 7.8 9.0 20.4 ---- 63 50 41 29 24 n 0 0 0 0 0 statistics: covar + plimnett tests rto.os r<o.oi tud siPEU (EXP .UNIT = ANIMAL) zosm ssa f r~ r -- SOSt'St SSd ) rftPLE 7 HEAN POPY WEIGHTS ( G) (CONTINUED 7) FEMALES pay jay jAY JAY Tift Y JAY JAY JAY 'irtY (i A Y 5 AY JAY :iay JAY ) *i Y JAY JAY iA Y AY JAY l AY lAY lAY iAY iftY lAY iAY lAY 0 16 30 SB 06 II4 1 43 170 190 226 254 2B2 310 338 366 394 422 450 478 504 534 562 590 618 646 674 702 730 758 CONTROL MEAN SEH N 52.8 92 * 7 119.9 151.4 144.7 178.0 183.3 188.4 189.6 193.9 199.0 202.9 210.2 209 2 211.5 212.1 207.9 210.1 214.0 213.0 218.6 220.7 225.3 226.3 231.7 237.0 235.8 232.5 232.9 0.5 0.8 0.9 1 .1 1 .3 1 .4 1.5 1 .6 1 .6 1 .7 1.6 1.6 1.7 1.7 1.8 1.9 1.9 1.9 2.1 2.0 2.3 2.5 2.8 2.5 2.7 2.7 2.7 2.7 3.0 100 100 1 00 100 1 00 100 1 00 100 100 1 OO 1 00 100 9V 99 99 99 99 99 99 9B 9B 96 95 94 93 90 89 86 BO 0.017 MEAN HG/KG SEH N 52.9 92.5 120.2 153.4 166.2 1 75.2 103.4 107.2 192.9 192.7 201.0 203.7 209.7 20B . 1 212.4 210.0 207.6 210.3 212.4 213.1 217.6 219.0 222 7 227.0 231 .9 236.0 234.4 232.3 234.6 0.5 0.7 0.9 1 .1 1 .4 1 .5 1 .7 1 .7 1 .B 1 .B 1 .7 1 .9 2.0 2.0 2.1 2.1 2.1 2.1 2.2 "? ** 2.5 2.5 2. 7 2.B 2.9 3.0 3.0 3.0 3.2 100 1 00 100 100 100 1 00 J 00 100 100 100 100 100 100 100 100 100 100 too 99 98 VB 96 96 94 95 93 92 90 87 0.17 HG/KG MEAN SEH N 51.5 90.1 120.1 1 5 3.6 168.6* 170.5 1 e4.0 190.6 190.0 195.6 201.8 205.9 210.8 210.8 211.6 212.9 209,0 213.4 215,4 216.0 218.4 221.8 226.5 230.3 235.0 24 1.0 240.0 238.8 240.2 0.5 0.7 0.9 1 .0 1 .2 1 .3 1 .5 1 .5 1 .6 1.7 1 .6 1 .6 1 ,7 1.9 1 .8 1.9 1 .9 1 .9 2.0 2.0 ?.7 2.2 2.4 2.5 2.6 2.7 2.8 2.7 2,9 100 1 00 100 100 100 1 00 1 00 100 100 100 100 100 loo 100 100 99 99 98 98 97 97 97 96 96 93 91 89 e6 81 TATISTICS CD'JAR 4 PUNNETT TESTS ' F'-TO 05 ' ' F- .0.01 TWO SI HEP 1 7 HG/KG HEAN SEH n 51.2 88.5* 117.9 152.0 147.8 178.9 187.8 194.3* 194.0 195.4 203.3 205.5 212.3 213.1 214.6 217.3 212.7 217.0 217.9 217.0 223.3 22B . 0 232.9 240.8 245.1 242.8 237.0 237.4 0.7 1 .2 1 .3 1.5 1 .8 2.0 2.1 7.2 2.4 *1 ^ *1 2.4 2.6 2.0 2.8 3. 1 3. 1 3.0 3.2 3.3 3.3 3.6 3.7 3.5 3.8 4.0 4 .2 4 .4 4.0 1.2 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 50 49 19 4B 4B 48 46 44 41 44 44 38 36 CONTROL HEAN SEH N 52.8 90.4 * * 117.4 160.0* * 167.3 176.3 182.8 189.9 193.0 195.8 205.2 206.7 212.3 210.0 211.5 213.2 209.7 211.2 215.9 210.8 219.1 219.3 221.9 223 * 2 227.7 226.8 * ^74 . n * 21' .8* . 21 y. o *4 0.4 0.B 0.9 I .2 1 .1 1.5 1 .5 ! .6 1 .4 \ .L 1 .8 i. e i .? 1 *D l .o ! .9 1.B 1 .9 2.0 2,0 2.o 2, 1 2. 1 *1 *> 1 -) 2.4 2.6 2 . ) 2.5 1 00 1 00 100 1 00 100 100 100 100 1 00 1 00 1 00 1 00 1 00 100 1 00 100 100 1 00 100 1 flA 1 Ov 100 99 98 ?B 9a 91 86 (EXE. UNIT - AN I HALJ r~ ~~~] r ^ 1 i-----1 f--i " "i ^ ' --] TABLE 7 MEAN POI'Y UEI GUTS (G ' (CONTINUED 3) F E HALES CONTROL MEAN SEN N 0.017 HG/KG MEAN SEN N 0.17 NG/KG MEAN SEN H 1.7 HG/KG MEAN SEN N CONTROL HEAN SEH N PAT 7B6 PAY e i -i PAY 8 4 2' PAY 870 PAY B9B PAY 926 PAY 954 PAY 982 PAY 1010 PAY103B P A Y 10 4 2 230.7 224.0 T'J'I t "J 217.9 216.6 212.3 211.1 212.7 212.4 214.9 2.8 2.6 2.7 2.9 2.9 2.9 3.0 3.3 3.4 3.6 3.3 BO 74 72 70 66 59 54 43 36 23 21 230.7 227.0 224.0 m 7 221.0 217,7 212.8 214.4 216.0 215.1 215.2 3.2 3. 1 3.4 3.6 3.9 3.8 4.3 4.6 5. I 5.7 5.7 B5 7B 69 65 59 49 41 34 28 23 23 235.4 232.1 231.8 227.3 223.3 220.8 215.1 212.1 221.1 21B. 4 217.7 2.B 2.7 2.8 2.9 3.2 2.9 3.0 3.8 3.5 4,1 4.2 81 78 74 70 66 55 4B 42 33 25 25 232.9 227.9 227.4 227 . 1 227.0 224.6 217.2 194.2 20 8.6 214.2 Op l' 3.7 3.7 4. 4 4 .2 4.9 5.4 5.5 4.8 3.7 3.1 3.8 35 34 31 29 26 nn 19 14 7 4 4 209 . B 1 207.7 203.1 * * 203.3 ' * 200.6*1 217.6 ----- 2.8 2.B 3. 4 3.7 4.0 4.3 -- -- -- D4 77 65 50 42 8 0 0 0 0 0 STATISTICS! COVAR + BUNNETT TESTS F<0.05 ' F1 ' 0.01 TIJO SIPES (EXF'.UNIT = AMI HAL ) t i POZPZl SSd sozm S'Sd .. ^ ] 1 >> I*L C 8 <1 ' V.M IK I III', Ulhl HE AN KOHY HEIGHTS (G>. MALES HAY HAY HAY HAY HAY HAY HAY HAY HA l HAY HAY HAY HAY HAY HAY HAY HAY DAY HAY HAY HAY 0 14 ;-e 56 BA 112 140 1 66 196 224 2 5n 280 308 336 364 392 420 4 4B 476 504 532 CONTROL MEAN SEH N 51.4 09.4 14 1.6 229. 276.4 312.1 336 . 1 360.4 373.5 303.4 302.6 417.1 424.6 430.3 4 13.0 419.7 416.5 413.7 422,1 4 13.5 426.6 2.0 2.5 3. 1 4.6 4,8 5.1 5.4 7. 1 6.2 7.4 12.7 11.9 9. 4 10.3 9.7 8.1 6.6 10.1 10.8 9.8 11.6 10 10 10 10 10 10 10 t0 10 10 10 5 5 5 5 5 5 5 5 e- 0.017 HG/KU MEAN SEH N 53.4 94.0 146,3 232.8 200.7 325.9 354.7 374.5 3 B 4.2 3e5.5 395.4 425.5 434.3 439.9 440.0 436.0 432.6 434.0 421 .0 405.4 423.2 2.1 2.4 3.2 5.4 6.3 7.3 10.2 3. 3 9.7 12.2 9.7 0.5 12.2 13.2 13.0 11.9 10.4 10.9 0,0 10,2 9.2 10 10 10 10 10 10 10 10 10 10 10 5 5 3 5 3 5 5 3 3 3 0 MEAN 55.5 96.9 139.5 230.4 209.4 325.7 350.7 372.3 3 7 7.5 308.4 394 .2 393.1 392.4 400.1 395.6 391.0 379.2 382.4 381 .5 '374.9 395.1 statistics: A- 3VA + DUHMET1 TEST rs * T 0.05 R'.O, Oi HO/KG SEH H 1 .7 MG/EG ML All SEH H CON [RUL MEAN SEH 1.5 2.0 9.1 4.3 6.5 8.0 9.6 10.6 P.5 0.5 9.4 6.5 6.5 7.8 5.6 5.8 12.7 9.3 12.1 14.1 14.9 10 10 10 o 9 9 9 9 9 o 9 4 4 4 4 4 4 4 4 4 4 56.4 97.0 1 49,5 224.9 200.3 321.0 341.9 360.0 379.6 391.1 394.1 412.3 4 15.1 4 10.4 421,4 421 .0 413.2 417.9 421.8 400.7 422.7 1.0 1 .0 2.0 3.2 5.0 7^ 6.3 6. 1 4,9 5.5 7.0 10. i 10.0 0.2 0.9 0.4 0,0 8. 1 7.5 4,8 6.G 10 10 10 10 10 10 10 10 10 10 10 5 5 5 5 5 5 5 5 5 5 54,9 96.4 14 7, 7 218.1 246.0' 2 7 0.7,' ' 2 G 6.9 ' ` 306.5' ` 314.6' ' 337.8' ' 340.6' 352.3' ' 361.3' ' 364.1' * 373 . 1 372.9' 359.2' ' 356,0* ' 359.6 ' * 353.J' ' 369.2 ' 1.5 2.3 4. S. 4 9.0 ,2 9,& 9. 7 B.B 9.5 8. 1 16.3 16.7 16.3 14.9 15.6 18.1 16.9 16.5 15.7 1 '. 0 ....... ..... ____ ______ _ _ ............... .. ___ .. ............ 1VJ0 5 11>E Ci (EXE.UNIT - AN1HA1. i study no 6U ia-nomn siuur in kais wnn vui in mil mu TABLE 8 MEAN DODY UEIGHTS <G). (CONTINUED) FEMALES DAY DAY DAY DAY DAY DAY DAY DAY DAY DAY DAY DAY DAY DAY DAY DAY DAY DAY DAY DAY DAY 0 16 30 58 86 11 4 14 3 170 178 236 254 282 310 338 366 394 422 450 478 506 534 CONTROL HE AH SEH N 53.3 95.4 119.8 150,3 161.7 172.3 177.8 18' .0 185.9 187.7 195.3 195.7 197.3 198.7 200.7 203.1 198.8 201.4 208.0 20<?, 0 214.5 1 .4 1 .8 2.5 2.7 3.8 4.0 4.9 5.1 5.4 5.5 5.5 3.2 4.5 3.0 5.2 5.3 5.5 5.9 6,4 5.5 7.0 10 10 10 10 10 10 10 10 10 10 10 tr 5 5 5 5 5 5 5 5 0.017 HEAH HG/KG SEH H 54.3 92.2 119.3 154.5 168.9 179.5 185.3 193.3 193.9 195.9 204.8 209.8 214.3 213.2 220.3 216.5 215.6 217.1 219.1 225*2 226.6 1 .7 2.7 2.8 3.7 3.6 3.4 3.4 3.9 3.4 3.6 3.2 3.0 5.1 4.4 5.5 6. 1 6.4 6.8 7.8 7.9 9. 1 10 10 10 9 9 9 9 9 9 9 9 5 5 5 5 5 u 5 5 5 5 0. 17 HD/K 8 HEAH SEH H 54.7 95.2 123.1 154.4 166.3 178.5 18 5.9 190.7 191.0 196.7 201.3 195.1 202*2 199.5 198.2 198.7 199.6 201.0 200.7 'IQ'1 > 204.3 1 . 6 10 1 . 9 10 2.4 10 3. 1 10 3.3 10 4 . 1 10 4.6 10 4.9 10 4.2 10 5.0 10 4.3 10 7.2 5 8.3 5 6.9 5 8.1 5 8.4 j 8.1 5 7.5 5 8. 1 5 8.8 5 9.0 j statistics:1 AMOVA + PUNHETT TESTS P<0.05 * F<0.01 TWO SIDED i .7 MG/KG HEAH SEH H 52.5 87.2 116.2 147.9 162.7 175.6 182.1 195.9 188.9 191.0 199.4 198.3 202.3 201.7 201.6 204.0 199,5 204.2 205.2 205.8 209.4 1 .6 2.9 3.3 4.0 4.8 5.8 55 6. 1 6.0 5*A 6.1 11,3 11.9 10.5 10,4 10.6 10.1 11.0 11.2 11,3 12,4 10 10 10 10 10 10 10 10 10 10 10 5 5 5 5 5 5 5 5 COHTROL HEAH SEH 50 . A 85.7' 111.8 152.2 160.9 173.6 179.6 184.8 186.2 190.4 198.3 204.7 2 11.4 211.6 2 15.4 2 14.0 210.7 217,2 214.2 223.0 1.6 2. 1 2.4 2.9 4 4 4,9 4. 3 5.4 4 .B 5.4 5. 7 5.0 5.8 6.0 56 6.4 5.3 5,7 6.4 5. 1 6.0 (EXP. UHIT = AHIHAL ) 902m S'Sd P'~'' ! [--r r LQZWl S'Sb '1 n \ C1VO/TNO STUDY HO 125 CARCINOGENICITY STUDY IH RATS WITH VCM IN THE DIET TABLE 9 HEAN FOOD INTAKE Croup A B c D E Intended Intake levels of VCH (og/kg b.v./day KALES 0 0.017 0.17 1.7 0 Food Intake (g/ra t/day) in week 1 2 3 4 11 12 23 24 3S 36 8.9 10.3 12.3 14.4 16.1 16.1 16.5 16.3 15.3 16. 7 7. A 9.7 11.1 13.5 14.0 14.5 14.5 14.0 14.8 15.5 7.7 10.3 11.9 14.7 15.6 15.6 17.4 17.0 15. 5 16.6 7.8 10.4 12.1 14.5 14.7 15.3 17.4 17.0 15.9 15.5 B.2 10.3 12.4 13.5 12.5 13.3 12.5 11.9 13.3 12.9 47 15.7 14.6 15.8 15.5 12.3 48 17.0 15.3 17. 1 16.4 13.0 59 17. 1 16.6 15.9 17.0 12.5 FEMALES A 0 8.9 10.0 9.9 10.7 10.4 . 10.2 9.2 9.1 9.3 9.8 10.6 10.2 9.9 B 0.017 8.3 9.8 10.4 11.0 10.2 10.0 9.0 9.2 10.3 10.5 9.4 9.8 9.6 C 0.17 8.5 9.7 11.5 12.0 11.0 10.8 10.8 10.4 10.3 11.3 10.3 10.6 10.0 D 1.7 8.3 9.4 10.4 10.8 11.0 10.7 9.3 9.6 10.8 11.1 9.3 10.4 10.3 E 0 8.3 9.3 10.2 9.9 9.4 10.3 9.1 9.2 9.9 9. 7 9.8 9.9 10.0 I-------- r_ r-- CIVO/TNO STUDY HO 125 CARCINOGENICITY STUDY IN RATS WITH VCH IN THE DIET TABLE 9 KEAN FOOD INTAKE (CONT.) Croup A B C D E Intended Intake levels of VCH (of>/kR b.w./day HALES 0 0.017 0.17 1.7 0 Food Intake (g/ra t/day) In week 60 73 74 85 86 97 98 109 no 16.2 15.2 16.1 15.8 12.6 16.5 15.9 17.5 18.2 13.3 16.6 16.1 17.3 17.5 13.3 14.4 13.6 15.0 15.7 13.0 16.2 14.6 16.3 14.9 12.4 14.7 14.6 15.4 15.7 12.3 14.2 15.2 15.9 15.6 11.6 15.2 15.4 15.7 16.1 11.4 15.0 14.5 15.7 15.4 11.2 121 15.9 14.6 15.8 14.2 12.3 122 16.1 15.0 15.8 15.8 12.5 FEKALES A 0 9.7 10.4 9.0 8.9 9.7 9.7 9.6 10.1 10. 2 10.8 10.8 B 0.017 10.0 10.9 10.5 9.9 9.3 10.4 11.8 10.5 10.4 11.7 12.1 C 0.17 9.9 12.0 11.0 9.8 10.8 10.4 10.9 11.0 10.4 11.4 11.6 D 1.7 9.7 11.1 10.6 10.1 9.9 10.8 10.9 11.4 10.5 11.3 13.3 E 0 9.5 9.8 10.3 9.2 9.2 8.8 8.7 9.7 9.2 10.0 11.0 1 h- SQZtrtl 602t'|. ssy ClVO/TWO STUDY HO 125 CARC1HOGEH1ClTY STUDY IN RATS WITH VCM IN THE DIETS TABLE 10 FOOD INTAKE DURI G THE FOUR-HOUR FEEDING PERIOD Croup A B c D E Intended oral Intake levels of VCH (mg/kg b.v./day) HALES 0 0.017 0.17 1.7 0 Food J ntake during the feed!ng-perJod over periods of one hour* ^ expressed as g/rst expressed as Z--tnge of total food consumed per rat over the 4-hr period 0-1 ' 1-2 2-3 3-4 TOTAL 0-1 1-2 2-3 W+ U.62) 0.87 9.9 0.66 10.5 0.58 10.7 0.90 8.8 0.66 2.7 0.29 3.2 0.29 3.0 0.27 3.6 0.34 2.6 0.24 2.0 0.27 2.3 0.11 2.5 0.23 2.3 0.25 1.7 0.17 1.9 0.31 1.8 0.17 1.7 0.16 2.0 0.29 1.6 0,15 18.2 0.62 17.2 0.71 17.7 0.60 18.5 0.95 14.6 0.69 64 15 11 10 58 19 13 11) 59 17 14 in 58 19 12 11 * 60 IB 11 11 overall A B C D E FEMALES 0 0.017 0.17 1.7 0 6.6 0.32 6.5 0.44 6.5 0.30 7.1 0,46 6.2 0.36 2.1 0.19 2.4 0.23 2.3 0.20 2.3 0.21 2.1 0.13 1.7 0.10 1.6 0.19 1.9 0.18 1.8 0.16 1.5 0.14 1.7 0,26 1.5 0.18 1.6 0.24 1.6 0.17 1.6 0.18 12.1 0.40 12.0 0.53 12.3 0.30 12.8 0.62 11.4 0.31 60 18 12 10 55 17 14 14 54 20 13 13 53 19 15 13 55 18 14 1 1 54 19 J 3 14 overall ^ Food Intake figures are the means of 11 different determinations. ^ Standard error'of the mean 54 19 )4 l_i 11 TABLE 11 MEAN MAEHAT OLOGI CAL FINDINGS RECORBE0 AT HAY 94.1B5 ANH 367 MALES . CONTROL MEAN SCM N 0.017 MG/KG MEAN SEH N 0.17 HG/KG MEAN SEH N 1.7 MG/KG MEAN SEH N CONTROL MEAN SEH N HAY 94 THROMHOC t10E9/L) 9B2. 27. 10 1036. 33. 10 1000. 23. B 996. 20. 10 B66. 50. 9 HAY 1B5 THROMHOC ( 10C9/L> 09(1. 17. 10 915. 43. 8 060. 45. 9 825. 51 >. 10 B79. . 31 . 10 HAY 367 THROMHOC ( 10E9/L) 946 . 23. 5 055. 43 . 5 B95. 21. 4 930. 63. 5 B72. 32. 5 HAY 9 4 PTT (SEC> 55,0 1.7 10 52.9 1.5 10 53.0 n " ? 50.0 1.3 10 54.2 2.3 0 HAY 105 PTT (SEC) 36.0 0.0 10 35.2 0.4 B 37.2 0.6 9 37.0 0.7 8 39.0 0.5 10 HAY 367 PTT (SEC) 4 9,3 1 .4 5 50,1 2.7 J 50.5 0.6 4 50.2 1.0 J 32 * 5 1.2 3 STATISTICS! MANN'WHITNEY U--TEST ' P<0.05 THROMHOC = 1HROMHOCYTE PTT = PROTHROMBIN TIME ** P<0.02 " P-:0.002 TUO SIOEH (EXP,UNIT - ANIMAL) oizpi s'sy r~' r ; r -? i--: t--i r~i r --i , r~------r --i r~ - r----- i ----i r..... t ------ , uzm ssa ----- 1 - - "I - SjlUUl NU. TABLE 11 ll-| ICH* 1 ll ~ .. -O 1 OO 1 *41 >1 * ....................... MEAN HAEMATOLOGICAL FINIilNGS RECORBEH AT HAY 97.104 ANO 366 F EHALE S CONTROL MEAN 5CM W 0.017 HG/NG MEAN SEM N 0.17 HG/KG MEAN SEN N 1.7 MG/KG MEAN SEM N CONTROL MEAN SEM N OAT 97 THROHBOC (10E9/1> 977. 19. 10 915. 27, 9 904 . 14 . 10 963 34. 10 943. 20. 10 HAY 104 THROMHOC (10E9/L> 972 . 27. 10 1032. 34 , 9 971. 23. 9 1018. 22. 9 1010. 29. 10 HAY 36A 11IR0IIH0C ( 10E9/L > 879. 26. 5 930. 106. 3 904 . 24 . 4 906. 20. ` 4 929. 131 . 3 HAY 9 7 PTT (SCO 41.6 0. El 10 40.2 0.0 9 43. 1 1.6 10 47.4 1 ' 2.1 0 51.3* * * 1.9 10 HAY 104 PI T (SEC > 33.2 0.5 9 32.8 0,4 9 33.0 1.0 8 36.2 1.7 9 37.O''* 0.7 9 HAY 366 F-TT i sn.` > 40.1 1.2 3 43.6 2.0 A 39.4 1.2 5 39.6 1 .7 5 39.9 1.0 4 . . ............. F'<0.05statistics: mann/whitney li-tcst * THR0MBOC => THROMBOCYTE F'TT = PROTHROMBIN TIME r-<o,02 '* 1X0.002 TWO SI OFH (EXP.UNIT = ANIMAL) 1 1 i | 1 r" ii CIVO/TNO STUDY HO 126 18-HONTH STUDY 1H RATS WITH VCH IN THE DIETS TABLE 12 HEAD HON-FROTEIN BOUND SULFHYDRYL ("GLUTATHIONE") CONTENT OF LIVERS OF RATS FED VCH FOR 9 AND IB MONTHS Croup A B C D E Intended exposure levels of VCH (mg/kg/day) 0 0.017 0.17 1.7 0 Non-protcin bound liver Sll In umol ftlutathlone/fc liver in months 9 18 9 18 HALES 7.58 7.52 0.34 0.38 7.32 7.57 0.38 0.47 7.38 7.33 (4) 0.29 0.28 7.30 7.63 0.22 0.22 7.31 7.44 0.36 0.38 FEMALES 6.05 5.58 0.34 0.63 5.66 (4) 5.69 0.18 0.14 5.95 5.95 0.29 0.31 6.57 6.13 0.36 0.43 6.52 6.30 0.58 0.29 Unde r each mean the standard error of the mean Is given. In brackets number of rats If not 5. UN UJ i l. i1. . r l i L. r L JJ 99 CO pit CO Pa ro CO r** i r i Lj rr r i i (" I W_t cr> r*' j L !f '' u ff - L r > u. L ' :vo/tno - I' TJDY NO 125 CARCINOGENICITY- STUDY IN RATS WITH VCM IN THE ________ _D_IE_T__ _____ __ _ __________ '.ic.r. 13. macroscopic pathology ;ite and type of observations Initial number of animals effective number of animals 10DE OF DEATH: a. found dead b. killed ir. extremis 0. terminal sacrifice UJTOLY'SIS/CANNIBALISM: a. slightly autolycic b. partially lost c. completely lost SKIN / SUBCUTIS 1. tumour or suspected of tumour 2, alopecia j, edema 4. haemorrhage 5. injury AXILLARY LYMPH NODES 1. tumour or suspected of tumour 2. enlarged 3. reddish PREPUTIAL -/CLITORAL GLANDS 1. tumour or suspected of tumour 2. evidence of inflammation 3, enlarged 4. small 5. cyst MAMMARY GLANDS I. tumour or suspected of tumour 2. evidence of secretory activity 3* haemorrhage ABDOMINAL CAVITY 1. tumour or suspected of tumour 2. ascites Incidence of observations MALES mg VCM/kg b.v /day 0 . 01 ) 0. 1 ) i. 7 0 FEMALES mg VCM/kg b.v. /dav 0 017 0. 17 1 .7 0 100 100 100 50 100 100 100 too 50 100 99 100 99 49 100 98 100 96 49 100 26 26 25 16 10 54 54 57 27 90 20 20 18 7 0 8 15 9 5 8 71 62 67 41 92 21 23 24 4 0 10 15 13 8 9 11 4 4 9 1 10i 10 364 2 6 064 25 204 1 0 57 93 3 53 3 2 4 1 7 2 3 10 9 9 4 8 232 1 0 1 1 1 00 00 1 00 00000 0020 1 0000 1 3 200 0 2200 0 1 0000 000 1 0 0 1 20 1 00000 352 1 2 7 6 11 2 2 01 1 00 00 20 0 00000 3 1 020 55 1 35 00 1 1 0 00000 0 1 000 8 14 7 3 3 39 25 28 18 23 12 6 8 2 6 53 35 41 21 29 00 I 00 00000 ]2 1 10 35320 685 26 55546 - a* ' ; : Y 1 j 1 1 JV.j/INO STUDY NO 125 CARCINOGENICITY STUDY IN RATS WITH VCM IN THE DIET `.'Ai.l.r. ! 2. MACROSCOPIC PATHOLOGY (CONTINUED I) R&S 134214 Sit*.* and type ot ob.svrvjtiuns SPLEEN 1. tumour or suspected of tumour 2. splenomegaly 3. atrophy 4. granular surface 5. snail 6. small nodule 7. dark ADRENALS 1. tumour or suspected of tumour 2. small (unilateral) 3. haemorrhagic 4. discoloured a. pale b. dark 5. spotted 6. enlarged a. unilateral b. bilateral KIDNEYS 1. tumour or suspected of tumour 2. granular surface 3. unilateral hydronephrosis 4. discoloured a. pale b. dark c. greenish d. reddish 5. cyst(s) 6. enlarged 7. small E. spotted STOMACH 1. enlarged/swollen 2. haemorrhage/erosion 3. rough inner surface 4. chickened cardial/fundic wall 5. thin fundic wall 6. trichobezoar Incidence of ubsurv.itions MALES mg VCM/kg b*w./duy 0 .017 0.17 1.7 0 l'UMALES mg VCM/kg b.w./dav 0 To P7 ' b .17 1.7* C~~ 0 10 10 8 14 7 3 2 0 00 0 1 2 0 110 10 0 11 0 00 0 1 0 00 0 0 0 10 0 3 42432 0 0 0 10 00 1 I C 00 2 00 0000 0 0 10 0 0 7 52 0 1 1 0 10 1 0 00 0 0 101 I 13 3 6 10 2 9 8 7 12 6 28 21 3 22 21 5520 1 110 0 0 1110 0 2 0 12 1 42747 26 25 27 14 23 21 9 6 115 47 14 0 23 1110 9 1 I0 5 27 10 2 112 0 00 5 44 3 24 0 00 10 0 00 18 I0 23 01 00 01 24 02 00 01 0200 52 20 3230 3 110 0 10 0 3 3 13 0000 1 430 112 0 0 10 0 0000 0 0 0 0 1 0 0 I 00 1 4 16 3 16 5 4 10 0 0 0 10 0 0 0 10 n 1I n 6 1 0 0 0 * 14 06 00 10 00 32 12 00 00 00 0 2 0 0 0 L i j' 1 r- I 1 1 }i 1 1 33 i So 1/7 < OJ t 4b. ro cn . I L 1 *. CIVO.'TNO STUDY NO I'5 CARCINOGENICITY STUDY IN RATS WITH VCN IN THE DIET II. il.i'Hiim :>>' 1 niOLOOV (CONTINUED ?.) - 00 Site and type of obscrvjlions SMALL INTESTINES 1. Humour or suspected of tumour 2. prominent Peyers's patches 3, enlarged/swollen 4, haemorrhage 5. reddish 6. filled with watery fluid COECUM 1. tumour or suspected of tumour 2. enlarged/swollen 3. small 4. filled with watery fluid COLON 1, tumour or suspected of tumour 2. enlarged/swollen 3. filled with watery fluid 4. accreted to the cervix MESENTERY 1. tumour or suspected of tumour 2 ed ema 3. dilated blood-vessel 4. thrombus in blood-vessel 5. haemorrhage PANCREAS 1. tumour or suspected of tumour 2. discoloured a. pale * b. dark 3 edema 4. spotted c. reddish 5. small haemorrhages MESENTERIC LYMPH NODES 1. tumour or suspected of tumour 2. enlarged 3. small 4. edema 5. haemorrhagic Incidence of observations MA1.ES mg VCM/kg b.v./day 0 .017 0.17 1.7 0 FEMALES mg VCM/kg b.v./<Jav 0 .017 0.17 1 .7 0 02000 34000 333 1 0 1 22 1 0 1 0 1 00 00 1 0 1 00000 1 00 1 2 3 1 000 0 I 000 000 0 0 0 00 00 00 1 00 857 5 3 0 1 000 0 0 .0 0 1 00000 33000 0 00 0 0 0C000 1 0000 02 2 2 0 0000 1 00000 1 0000 1 0000 00 0 0 0 00 1 00 0 1 200 004 00 443 1 0 00 1 00 000 0 0 2 24 03 1 0000 54 1 1 3 00000 00 1 00 2 1 1 00 2002 1 4030 1 1 0 1 00 0 1 1 00 30000 00000 44 24 5 000 1 2 1 2202 000 1 0 00000 00000 1 0000 42200 33 1 1 1 01 1 0 1 00 2 03 1 0000 4423 2 6 1 300 2121 1 2 1 1 03 0000 1 -- i r _ 1 ; L j > ; ! 1r- j 1 r 1 L i L t !1L_, f. L r L UJVO/TNU study no 1:5 carcinogenicity study in rats with vcm in the diet TAIW.F 1 3. MaCKOSCOI'II pathology (CONTINUED 3) Site and type ot observations 6. discoloured a. pale b. dark c. greenish d. reddish URINARY BLADDER 1, tumour or suspected of tumour 2. haemorrhage 3. enlarged/svollen 4. calculus/concrements SEMINAL VESICLES 1. tumour or suspected of tumour 2. evidence of inflammation 3. enlarged 4. atrophy coagulation glands 1. tumour or suspected of tumour 2. evidence of inflammation 3. enlarged 4. atrophy PROSTATE 1. tumour or suspected of tumour 2. evidence of inflammation 3. enlarged 4. atrophy TESTES ). tumour or suspected of tumour 2. atrophy a. unilateral b, bilateral 3. enlarged (unilateral) 4. cryptochismus EPIDIDYMIDES K tumour or suspected of tumour 2, evidence of inflammation 3. atrophy 4. haemorrhage Incidence of observations MALES mg VCM/kg b-w,/day 0 . 01) 0.17 1.7 0 1 00 1 0 1 0 1 00 00000 00 200 FEMALES mg VCM/kg h . V* / dav 0 . 017 0.17 1.1' 0 0 000 0 0 000 0 00 1 00 002 1 1 001 00 1 2000 1 2002 00 1 00 1 1 000 00200 32 1 00 00000 0 1 000 2 1 1 00 1 0000 4 5 10 3 27 ---- -- -- 1 1 000 1 0000 00000 3 5 9 3 27 ---- -_ --- 0530 1 1 2000 -- 1 J 00 1 -- 4 1 2 1 8- - - ---- 304 14 10 IS 10 9 6 202 1 01 00 57 5 22 10 12 ----- ----- 1 0000 00 I 00 2000 1 I 0000 -* - -_ --_ __ R&S 134216 ' , - - r f~ ! 1 i, , u. i r1 r DO Q CO _s CO rSo* i *sl i r l. r ` * L i- F T, L r; uvo nj study no 1:5 carcinogenicity study IN RATS W :th vex IN THE D15 T .'AIU.H 13. HACKO.NTOi'li PATHOLOGY (CONTINUED 4) - 36 - iitf and type of observations OVARIES 1. tumour or suspected of tumour 2. cyst (s) 3. enlarged 4. reddish UTERUS 1. tumour or suspected of tumour 2. hydrometra 3. evidence of inflammation pyometra 3. enlargcd/svollen -IVER 1. tumour or suspected of tumour 2. prominent lobular pattern 3. swollen/enlarged >. discoloured a. pale b. dark i. cyst(s) >. small small surface lesions (rosette, constriction) spotted granular surface 'HORACIC CAVITY . tumour or suspected of tumour . hydrothorax . haemothorax HYMUS . tumour or suspected of tumour enlarged . spotted ART . enlarged (atria) . thrombus . pale Incidence of observations MAKES FEMALES mg VCM/kg b e V 7IHT7" mg VCM/kg b . V , /dav 0 . 017 0.17 1.7 0 0 . 017 0.17 1 .7 0 ------- ---- ------ 64 23 5 8 ] 15 3 7 52 4 2 7 2 1 1 1 0 - - - - - 21 29 28 1 1 33 - --- - 21 1 32 --- - - 1 1 000 --- -- 02000 - - - - - 10 2 2 2 7 I 1 27 2 I 380 1 4200 01 3 1 0 245 1 0 3 2000 320 1 0 131 21 3300 1 000 1 0 2 1 5 4 0 17 14 22 33 3 0 1 1 02 002 00 1 01 01 5 682 3 2 00 2 0 3 000 1 2 1 622 0 1 200 00000 35830 48740 00000 262 1 0 5 1 000 1 1 000 0 1 000 1 0000 00020 1 000 1 00000 14 19 16 C 3 3750 00 1 00 67 50 1 454 00 00000 i t i * -- i JlVu.'TNO STUDY HU 125 CANCINOCENJCITY STUDY IN RATS WITH VLM IN THE DIET i.Mu i'j. riACisDSi one pathology (CONTINUED 5) -V- R&S 134218 Site and type of observations MEDIASTINAL LYMPH NODES 1. tumour or suspected of tumour T. enlarged LUNGS 1. tumour or suspected of tumour 2, evidence of CRD*) 3. spongy 4. spotted 5. haemorrhage 6. discoloured a. pale b. dark c. reddish 7. atelectasis (partially) 8. pleuritis TRACHEA 1. filled with blood AORTA 1 . dilated 2, calcified DIAPHRAGM 1. tumour or suspected of tumour 2. spotted surface CERVICAL LYMPH NODES 1. tumour or suspected of tumour 2 enlarged SUBMAXILLARY SALIVARY CLANDS 1 a tumour or suspected of tumour 2. edema 3* discoloured a. dark b. reddish 4* enlargvd/swollen Incidence of observations MALES mg VCM/kg b ,V. /day 0 . 017 0. 17 1.7 0 FEMALES mg VCM/kg b , V, /day 0 . 017 0,17 1 .7 0 3 I01 0 I " 2 2 O' '1 00 0 ' '2 00 f'2 0 o' 0!0 4 64 6 8 11 J8 J 8 26 1 04 1 20 1 00 I31 1 24 000 01 5 72 I 50 96 01 42 00 00 00 02 000 4 3 10 5 54 20 17 9 00 1 564 000 020 1 03 1 00 01 9 67 3 36 64 01 02 00 00 22 01 0 I 000 00 00 0 599 1 0 1 0000 52 1 1 2 0 0000 1 000 0 0 00 1 0 0J ]01 0 000 0 1 0000 0 1 1 00 0 000 0 0000 0 1 1 1 00 121 1 1 ] 0000 00 1 00 1 ] 000 0000 1 0001 0 00 000 0 1 00 0 0 0 00 0 * CRD * chronic respiratory disease I r~ JO B CO --4 CO ho ___L CO l r~ i i r ! 1r i_ ' L. .- UlVu/TNO STUDY NO I r5 CARCINOGENICITY STUDY IN RATS WITH VOi IN THE DIET Y' I'J. :lm NOSCiX'I l'.''THf1LO"Y (CONTINUED 6) - faC. - hili- ,nuJ type ol ul);.rrv.il ioui SUBLINGUAL SALIVARY CLANDS 1 a tumour or suspected of tumour 2. enlarged/swollen 3. haemorrhage 4. edema 5. discoloured a- pale b* reddish PAROTID SALIVARY CLANDS 1. discoloured, pale 2. spotted 3, enlarged THYROID I, tumour or suspected of tumour 2* enlarged 3. discoloured a* pale b. dark c, reddish PARATHYROID I. enlarged EYES 1, haemorrhagic scabs around eyes 2. evidence of inflammation 3. protruding 4. pale 5. opaque ERAIN 1. tumour or suspected of tumour 2. haemorrhage 3. a pale spot MENINGES 1, haemorrhage Incidence of observations HALES mg VCM/kg b ,W. /day 0 . 017 0.17 1.7 0 FEMALES mg VCM/kg b . w / vluv U a 017 0.17 1. 0 -- ] 0000 0 I 000 1 0000 0 1 1 00 1 000 0 00000 00000 00000 00000 00000 00000 0 1 000 3 00 1 0 000 I 3 0 1 000 00000 000 0 0 1 0000 263 1 0 30 1 10 1 0000 2 1 30 1 00000 1 2000 0000 1 00000 0 1 00 1 00 1 00 1 0000 00000 2 2024 02 1 1 1 1 6623 2 2 10 1 3 33002 6 9 9 2 21 8 6 6 2 0 12 12 3 7 3 13 1 13 7 9 16 2 14 3 S 0000 0 01 1 10 000 1 0 1 0000 00000 00 000 0020 1 0 0 000 f. < i VI I ' ! M -.nr. ::o i. j cauoinikjkn!ci ty studi is kats utii va: is the diet 1 1 1 3. "v`i:;\v Ol'l i' P.'Tiinl.iiCY (CONTINUED 71 <>! SjU' ,iud type of observations PITUITARY 1. tumour or suspected of tumour 2. haemorrhagic (cyst) 3. discoloured a. pale b. dark 4. enlarged/suollen NOSE 1, tumour or suspected of tumour 2. haemorrhagic scabs around nose ORAL CAVITY I. malocclusion of incisors EXORBITAL LACHRYMAL GLANDS 1. pale 2. granular appearence ZYMBAL's GLANDS 1. tumour or suspected of tumour 2. enlarged 3. evidence of inflammation i. haemorrhage ABDOMINAL/LUMBAL LYMPH NODES 1. tumour or suspected of tumour 2. enlarged ABDOMINAL WALL I. small cysts rail 1. ulcer 2. abscess EAR DUCT >. compact debris 2, evidence of inflammation -1MB5 1, tumour or suspected of tumour 2. abscess 1. deformed Incidence of observations MALES mg VCM/kg b-v./day 0 .017 0.17 1.7 0 FEMALES mg VQl/kg b.w./Jnv 0 .017 0,17 1.7 0 B 9 10 5 3 12 3 11 3 1110 I I 00 I 10 2 0 1 13 13 17 10 51011 4 24 0 0 00 1 I0 0 C 0 5 2 0 I 1 00 66 00 0 2 11 00 43 3 2 00 00 53 2 11 2 10 0 1 IS 12 10 8 13 69 6 17 00 00 00 00 0 0 0 I 0I 0 0 0 00 0 0 0 00 1 0 0 00 0 0 0C 0 10 10 00 0 0 0 10 0 0 0 0 0 110 0 0 2 10 0 0 0 0 10 00 0 0 0 0 0 0 0 0 0 10 0 0 0 2 11)0 0 0 00 0 00 0 0 0 0 10 0 0 0 10 0 0 0 10 0 0 00 00 00 00 0 0 2 0 00 0 0 0 00 1 4 2 10 1 00 00 1I 00 0 0 10 10 0 | j I R&s 134220  i:: lovrs i.'frn vc; in the nji.r (continiieo 5' MISCELLANEOUS 1, emaciaied 2* deformed spine 3, deformed dorsal vertebrae Incidence of nhservacion.; a-,", VITi/k)- |,.w.7`l;iy "itt Vai/k.'T, /.! i 0 .III 7~~t J~.~l i__ l',~7 0" ~(i ~Q| 7 0. 17 Y - * IS 19 13 00 17 7 00 10 48 0 0 32 22 26 1 4 55 0 I 000 0000 0 R&S 134221 ClVO/TNO STUDY NO 125 CARCINOGENICITY STUDY IN RATS WITH VCM IN THE DIETS TADl.E 14 TYPE AND INCIDENCE OF HI ST0PATII0E0C1CAL CHANCES IN THE LIVER Type of changes^ Number of animals examined Incidence of changes Males Females mR VCtl/kR b.w,. /day mg VCH/kg b.w. /tiny 0 0.017 0 .17 1.7 0 0 0,017 0.17 1.7 99 99 99 4 9 100 9B 100 96 49 0 99 1. Foci of cellular alteration a. clear cell foci I one or a few 11 several b. basophilic foci I one or a tew II several c. mixed cell foci I one or a few d. eosinophilic fori X one or a few All type of foci: Total Number of foclbearlng animals 12 8 00 42 00 01 11 ------------17 12 16 12 8 16** 0 3* 54 00 3 8* 00 09 00 22 06 21 " 15 30 00 5 19 15 23*** 5* A 19 5 3 13*** 1 0 0 6*** 0 20* 26*** 19* A A 4 1 020 6 u 8* 0 0 1 10*** 2 32 34 58 7 27 31* 32*** 7** i n tr ZZPl SSU CIVO/THO STUDY NO 125 CARCINOGENICITY STUDY IN RATS WITH VCM IN THE DIETS TABLE 14 TYPE AND INCIDENCE OF HISTOPATHO LOG ICAL CHANCES IN THE LIVER - CONTINUED 1 Type of changes 0 2. Neoplastic nodules a. one b. a few 3. Hepatocellular carcinoma 0 0 0 4* Angiosarcoma 0 5, Liver-cell polymorphism a. slight b. moderate 27 4 c. severe 1 6. Prominent sinusoidal cells a. slight 1 b, moderate/aarked 1 7. Cysts a. one b. s feu c. many 1 4 0 6. Bile duct prollferstIon a. slight b. moderate c. severe 37 9 2 Incidence nf changes Hales Females mg VCH/kg b.u., /dsy mg VCH/kg . /day 0.017 0.17 1.7 0 0 0.017 0.17 1.7 t 0 0 1 00 1 0 0 200 0 0 0 3* 0 1 0 0 0 1 00 0 1 9* * * 0i 13 02 0 0 0 0 0 23 26 19 12** 46 41 4 7 10** 1 14 13 1 1 302 3 49 23 38 8 15* 15 4 g*** 3 1 1 01 4 9 2 1 10 3 6 3 i6 5 20 0 1 00 3 2 4 3 4 1 11 11 0 0 00 3 4 4 11 12 7 2* 9 24*** 1 26 35 14 21 * * 27 23 9 8 3 1** 11 6 0 0 01 1 2 36 18 14* 10 2 3* 1 14 r~ r' i-- r- i t-- CIVO/TNO STUDY NO 125 CARCINOGENICITY STUDY IN RATS UIT1I VCH IN THE DIETS TABLE 14 TYPE AND INCIDENCE OF U1STOPATUOLOCICAL CHANCES IN THE LIVER - CONTINUED 2 Type of cliangea ___________________________________ Incidence of changes Kales Ferns 1 cs mg VCH/kg b.w./day rag VCM/kg b.w./day 0 0.01? 0.17 1.7 0 0 0.017 0.1? 1.7 0 9. Cholanglof1brosla a, slight b. moderate 33 19* 20* 15 23 16 13 17 15 2* * * 2 b 8* 21 b 2 u 0 0* 10. Periportal fibrosis a. slight b. moderate 1 2 2 350 0 0 0 1 2 0 0 020 0 0 0 0 11. Perl porta 1/cent rolobular / Infiltrates of mononuclear cells a. slight ba moderate c. severe 14 10 8 3 7 3 2 1 2 4 1 1 1 000 0 2 00 0 1 1 000 0 0 00 12. Foci of RES-cella occasionally Accompanied by a few necrotic hepatocy tes a. one or a few 18 15 27 5 12 39 28 42 12* 2 5* i b. several 5 8 3 01 6 3 5 03 i 11. Slight hacipji t opo 1 e t J c activity 2 0 0 01 1 0 3 0O vzzvzv s*y szzvzi s'sa CIVO/TNO STUDY NO 12S CARCINOCENICITY STUDY IN RATS WITH VCM IN THE DIETS TABLE 14 TYPE AND INCIDENCE OF HISTOPATHOLOCICAL CHANCES IN THE LIVER - CONTINUED 3 Type of changes 14. Single cell necrosis a. slight b,, ooderate/aevere 15. Focal necrosis a. slight b. moderate c, severe lb* Centrolobular liver-cell degeneratIon 17. One or a feu foci of degenerated hepatocytes 18. Haemorrhagic areas 19. Pellosls-1Ike changes 20. Small granuloma 21. Vacuolization of hepatocytes mainly a. periportal I slight 11 moderate 111 severe ____________________________________Incidence of changes Kales Females rog VCM/kg b.w./day mg VCM/kg b.w./day 0 0.017 0.17 1.7 0 0 0.017 0.17 1.7 2 3 1 04 2 3 13** 5* 2 1 0 001 3 4 2 11 3* 8 2 4* 4 4 6 6 1 2 1 003 1 2 0 1 0 0 203 0 1 0 1 3 3 1 03 2 2 0 10 10 01 00 7* 3 0 1 1 O00 0 201 0 002 2 00 0 10 1 00 0 00 3 6 11* 3 4 24 22 29 4* 0 2 5* 0 0 3 2 ft 0 0 1 1 00 0 0 00 0 9* 2 3 3 1 0 0 0 <1 0 6*** 1 0 I--------- (------ r| --- ^ | CIVO/TNO STUDY HO 115 CARCINOGENICITY STUDY IN RATS WITH VCH IH THE DIETS TABLE 14 TYPE AND INCIDENCE OF H1STDPATHOLOCICAL CHANCES IH THE LIVER - CONTINUED 4 dif fuse 1 slight 11 noderate 111 severe cent rolobular 1 slight Kales Fetnn 1 e s mR VCM/kfc b.v,. /day mfc VCM/kfi b.w,. /day 0 0.017 0.17 1.7 0 0 0.01 7 0.17 1.7 0 1 0 1 00 0 0 0 0G 1 0 2 00 2 0 000 0 2 0 00 1 0 002 0 1 0 00 0 0 0 00 Specific hepatocellular lesions were classified according to Squire and Levitt (1975), * P < 0.05; ** P < 0.01; *** P <0.001, according to the Fishers' exact teat (one tolled). 9ZZPZI SSb r~1 r izzpzi ssa C1VO/TNO STUDY NO 125 CARCINOCENICITY STUDY If) RATS WITH VCH IN THE DIET TABLE 15 TYPE AND INCIDENCE OF TUMOURS OF THE MAKHARY GLANDS AND INCIDENCES OF ABDOMINAL MESOTHELIOMAS Site and type of tumours Incidence of t umours Males Fenrn les mg VCH/kg b.w,. /day 0 0.017 0.17 1.7 mg VCM/kg b. w * / d a y 00 0.017 0.17 1.7 0 Humber of animals examined 99 99 99 A 9 100 98 100 96 48 9 fi Hammary elands " Ho of tumour-bearing animals 1. Adenoma a. single b. two 2. Intraductal papilloma 3. Fibroadenoma a. aingle b. two c. multiple 58 Q0 00 00 01 00 00 3 0 3 41 21 A * * 28* 21 20*' 0 0 06 4 2 52 0 0 01 1 1 00 0 0 00 0 0 10 0 0 0 25 10** 15 14 11* 0 0 02 3 2 24 0 0 03 1 1 11 '" ] 4 m ri r r i r~i CIVO/TNO STUDY HO 125 CARCINOGENICITY STUDY IN RATS WITH VCH IN THE DIET TABLE 15 TYPE AMD IHCIDEHCE OF TUMOURS; OF THE MAMMARY GLANDS AND INCIDENCES OF ABDOMINAL MESOTHELIOMAS CONTINUED Site and type of tumours 4. Fibroma a. single b. tvo c. multiple 5, Adenocarcinoma a. single b. tvo Incidence of tumours Hales Females mg VCM/kg b.u.. /day VCH/Wg b.v./day 0 0. 017 0.17 1.7 0 0 0,.017 0.17 1.7 0 4b 00 00 01 10 2 0 31 k 1 22 1 0 00 0 0 00 0 0 01 0 0 00 0 0 03 0 5 l0 0 0 00 0 2 01 Abdomen 1. Mesothelioma 01 2 0 03 2 2 11 * P < 0.05; ** P <0.01; *** P < 0.001, according to the Fishers' exact test (one tailed) i i i "'i c * QZZWl S'SU /;-/ - /O - 4-- ( -- ( --r ( \. \ \. * un wu am) t H |t |t| 141 JV4 )4J JT) 441 (W 1)4 (fIlVM4/t<< ( Mtm Mt * (IIKIHlth MP# 4CMII MMr Ll4a>; ** tH'ih **,4|7 M/M t . I M/*4 *- 1.7 M/M (4 tit *M 4 m 7** au W|k NT( I* fK ltw< CMTK FIGURE I HIT KtHl rnuii fi&S 134229 (|l4/ll* |t*|T iMtfi ir> V/fCHI III * [tl4(iHil4t MM ICMII MM LtMil' * (Mim .>* Hl C* *.l M/l| * I.T 44/11 |> (MIMS FI CURE 2 r~-. i-. 1 i i. r" ! 1 1r 1 l r . r i k-- r1 L_ f' !. Lr i L L ANNEX I 71 - PERCENTAGE COMPOSITION OF BASAL DIET FOR RATS AND MICE soya bean oil meal fish me*! meat and bone scraps wheat (whole ground) maize (whole ground) brewer's yeast grass meal whey powder defatted bone meal salt with trace elements ) B-vitamin mixture ) vitamin ADEK mixture^) soya bean oil n 7 4 36 29.7 3 3 2 0.4 0.3 0.1 0. 3 3 100.0 Trace elements in salt MnS04.H20 ZnClj KJ FeS04.7HiO CoC12 * 6H2O CuS04.3H20 NaCI "} B-vitamin mixture Thiamin-HCl Riboflavin Pyridoxin-HCl Niacin Ca-pantothenate Biotin Folic acid Vitamin Sl2"PreP* Ground sucrose (0.1 Z) Vitamin ADEK mixture Vit. ADj-prilla (Farmix): 2230 IU vit. A, 750 IU vit. D3/g Vit. E-dry powder (Merck) 30 2 Menadion-Na~bieulphite (Vit. K3) Wheat starch Batch 2380 250 20 1250 20 400 45680 E E 50000 Batch 25 30 100 125 75 1.5 5 50 E 9588.5 10000 Batch 25000 800 27 800 26627 E g U1 H > added to 1 kg diet 41 mg Mn 12 mg Zn 1. 5 mg J 25 mg Fe 0. 7 mg Co 8 mg Cu added to 1 kg diet 2.5 mg 3.0 tag 10.0 mg 12.5 mg 7.5 mg 0.15 mg 0.5 mg 5.0 mg (- 5 (Jg Bj2) added to 1 kg diet 6339 IU vit. A 2112 IU vit. D3 4 5 mg 3 mg vit. K3 37 Co 03 03 -tA fO CO o annex CONTAMINANTS REGULARLY DETERMINED IN BASAL DIET 1. Contaminants in basal diet contaminant detection limit tentativ< maximum inorganic substances^mg/kgl arsenic cadmium lead mercury selenium tin potassium nitrate sodium nitrite 0.01 0.005 0.05 0.005 0 JJ1 1 25 5 0.25 0.05 1.0 0.05 0.5 25 500 10 ortjanochlorine compounds (mg/kg) HCB lindane (g-HCH) o-HCH B-ttCH heptachlor heptachlorepoxide a-chlordane ir-chlordane aldrin dieldrin endrin methoxychlor p,p'-DDE p,p'-TD o,p'-DDT p,p`-DDT RGBS 0.005 0.01 0.01 0.02 0.01 0.02 0.02 0.02 0.01 0.01 0.02 0.05 0.02 0.03 0.03 0.0< 0.3 0.02 0.1 0. 1 0.02 0.02 0.02 0.02 0.05 0.01 0.01 0.02 0.5 0.02 0.03 0.05 0.05 0.5 organophosphorus compounds (mg/fcgl chlorpyrifos diazinon dichlorvos fenitrothion malathion mevinfos parathion sulfotep 0.01 0,01 0.01 0.01 0.01 0.01 0.01 0.01 0.05 0.05 0.05 0.05 0.5 0.05 0.05 0.05 dithiocarbamates, in total, determined as carbondisulf ide_ Ong/kg)^ 0.5 1 .0 ferbam mancozeb aaneb nabaa thiram zineb DO ziram Bo CO CO -pa. to CO 4* ANNEX 2 (COST.) Contaminants in basal diet (cont.) contaminant - golvc^clic aromatic hydrocarbons (pg/kg) benzo(b)fluoranthene benzo(k)fluoranthene bento(a)pyrene fluoranthene indeno(1,2,3-cd)pyrene - nitroso comoounds (ug/ka) nitrosodlethylamine nitrosodimethylacine nitrosodipropylaaine nitrosomethylbutylamine nitrosomethylpropylamine nitrosomorpholine nitrosopiperidine nitrosopyrrolidine - raycotoxins (mg/kg) aflatoxin Bj - miscellaneous determinations oestrogenic activity (Tiecco test) urease activity (dpH) trypsin inhibitors (mg trypsin/g food) haeoagglutinins negative reaction at dilution detection limit 0.2 0.4 0.2 1.0 0.4 0.5-1 .5 0.3-0.9 1.5-4.5 1.5-4.5 1.0-3.0 3.0-9 1.5-4.5 1.5-4.5 0.01 ' /J - tentative maximum 10 10 10 50 10 10 10 10 10 10 10 10 10 o.os negative 0.3 e.o 1:200 ANNEX 3 R& s 134233 NUTRIENT OPPOSITION CT BASAL DIET TOR RATS Analyses of a 1000 kg batch produced Janjary 2A, I960 moisture 12.6 X crude protein (N x 6.25) 20.9 X crude fat 8.9 X crude fibre 3.6 X total ash 5.6 X calcium 0.89 X phosphorus 0.72 X magnesium 0.1A X potassium 0.75 X sodium 0.28 X iron manganese copper Zinc cobalt chromium selenium vitamin A vitamin 0 vitamin E carotene vitamin K3 5200 l.U./kg vitamin 1380 l.U./kg vitamin aa mg/kg vitamin 2.0 mg/kg niacin 1,6 mg/kg pantothenic acid folic acid choline biotin vitamin Bj^ 175 ppm 95 ppm 15 ppm 50 ppm 2 ppm 3 ppm 0.2A ppm 4.7 5.8 12.0 66 mg/kg mg/kg mg/kg mg/kg 15.5 mg/kg 1.25 mg/kg 1600 mg/kg 0.22 mg/kg 8 ug/kg lsoleucine leucine lysine methionine cystine phenylalanine tyrosine threonine tryptophan g/100 g 0.85 1.60 1.03 0.39 0.36 0.89 0.6A 0.81 0.21 g/16 g N A .1 7.7 5.0 1.69 1.75 A.3 3.1 3.9 1.01 valine arginine histidine alanine aspartic acid glutamic acid glycine proline serine g/100 c) 1.00 1.16 0.45 1.09 1.74 3.69 1.11 1.40 1.10 g/16 g N 4.8 5.6 2.2 5.3 8.4 17.9 5.4 6.8 5.3 ANNEX 4 NUTRIENT COMPOSITION OF BASAL DIET FOR RATS 75 Analyses of a 1000 k batch produced September 9, 1980. - moiscure 11.9 2 iron 135 ppm crude protein (N * 6.25) 22.2 I manganese 95 ppm Lr i crude fat 6.4 2 copper 16 ppm crude fibre 3.1 2 zinc 55 ppm total ash 5.4 2 cobalt 2 ppm 1C" calcium 0.90 I chromium 2 ppm i phosphorus 0.78 2 selenium 0.40 ppm magnesium 0.15 2 iodine 2.0 ppm H potassium 0.82 2 sodium 0.29 2 t-- 1 vitamin A 7000 I.U./kg vitamin 6.2 mg/kg i L _ vitamin D I.U./kg vitamin 5.4 mg/kg vitamin E f' i 1 carotene L. vitamin 55 mg/kg mg/kg 1.8 mg/kg vitamin B0. niacin pantothenic ac id folic acid 11.8 mg/kg 74 mg/kg 11.5 mg/kg 2.4 mg/kg choline 1600 mg/kg biotin 0.20 mg/kg r vitamin B^ 16.5 yu.g/kg g/100 g/16 g N g/100 g g/16 g ' isoleucine 0.98 4.4 valine 1.14 5.2 leucine 1.78 8.0 arginine 1.21 5.4 lysine 1.13 5.1 histidine 0.44 2.0 -* methionine 0.41 1.8 alanine 1.19 5.4 cyst ine 0.38 1.7 aapartic acid 1.93 8.7 phenylalanine 0.98 4 .4 glutamic acid 4.20 18.9 r tyrosine 0.69 3.1 glycine 1.12 5.1 threonine 0.85 3.9 proline 1.42 6.4 L. tryptophan 0.23 1.0 aerine 1.17 5.3 1 1_ -Xi 9 CO co 4* to CO 4* annix 5 NUTRIENT COXPOSITION Of BASAL DIET FOR RATS An*lyse of a 1000 batch produced February 27, 1981 moisture 12.6 : i ron 150 ppm crude protein (N x 6.25) 20.6 I manganese 90 ppra r-- 1 crude fat 6.1 I copper 17 ppm crudefiber j.8 : zinc 45 ppm total ash 5.6 I cobalt 0.6 ppm i calcium i.o5 : chromium 3 ppm L phosphorus 0.76 I selenium 0.6 ppm magnesium 0.13 I iodine 1.5 ppm r[ potassium 0.75 : i sodium 0.32 Z R&S 134235 vi camin A vitamin 0 6600 -1) I.U./kg I.'J./kg vitamin vitamin 7.5 mg/kg 5.7 mg/kg vitamin 57 Bg/ kg vitamin 12.1 Bg/kg carotene 1.6 g/kg niacin 55 mg/kg vitamin mg/kg pantothenic acid 14.7 mg/kg folic acid 5.1 mg/kg choline 1500 mg/kg biotin 0.25 mg/kg r vitamin 15.5 Ug/kg g/100 g/16 g N g/100 g g/16 g N r isoleucine 0.81 4.0 valine 0.95 4.7 leucine 1.52 7.5 arginine 1.13 5.6 lysine 1.03 5.1 histidine 0.46 2.3 methionine 0.38 1.9 alanine 1.08 5.3 cystine 0.33 1.6 aspartic acid 1. 74 8.6 phenylalanine 0.88 4.3 glutamic acid 3.70 18.2 tyrosine 0.63 3.1 glycine 1.13 5.6 chreonine 0.78 3.8 proline 1.46 7.2 tryptophan 0.19 0.9 serine 1.06 5.2 r I L not analyaed CIVO-rNO, June 1981 L. j t \ I V__ . * R&S 134236 ANNEX o NUTRIENT COMPOSITION OF Cl VO-BASAL DIET FOR RATS Analyae of a 4000 kg batch produced by Hope Farms on Sept. 9, 1981 moisture 11.3 A iron 220 ppm crude protein (N x 6.25) 20.2 I manganese 105 ppm crude fat crude fiber 6.9 3.9 A A A A copper zinc 18 ppm 55 ppm total aah 5.3 I cobalt 1 ppm calcium 0.65 A chromium 1 ppm 1 phosphorus . 0.7 I selenium 0.25 ppm magnesium 0.21 T iodine 3.0 ppm f potassium 0.8 A L. sodium 0.32 A A i vitamin A 5700 l.V./kg vitamin B^ 9.0 B>g/kg vitamin 0 _1) I.U./kg vitamin 7 ng/kg vitamin E 56 mg/kg vitamin Bg 12 ng/kg carotene 13 mg/kg niacin 70 g/kg vitamin K, < 3 mg/kg pantothenic acid 18.6 ng/kg folic acid 3.3 ng/kg i choline 1700 "g/kg l biotin 0.24 ng/kg r vitamin B^ 13 g/kg i is.. g/100 g g/16 g N g/100 g g/16 t_ isoleucine 0.81 4.0 valine 1.05 5.2 i leucine 1.60 7.9 arginine 1.13 5.6 -- lysine 1.07 5.3 histidine 0.49 2.4 me thionine 0.40 2.0 alanine 1.14 5.6 f cystine 0.34 1.69 aspartic acid 1.73 8.6 pheny1alanine 0.89 4.4 glutamic acid 3.51 17.4 tyrosine 0.65 3.2 glycine 1.07 5.3 rm threonine 0.78 3.9 proline 1.27 6.3 tryptophan 0.23 1.14 serine 1.01 5.0 ^not analysed C1V0-TN0, June 1981 L [ R&S 134237 ANNEX 7 NUTRIENT COMPOSITION OF CIVO-3ASAL DIET FOR RATS Analyses of a 4000 kg batch produced by van Eck, Cothen, or. Febr. 9, 1982 moisture 11.3 I iron 315 ppm r~ crude protein (N * 6.25) 19.8 I manganese 95 ppm \ crude fat 6.2 Z copper 19 ppm V . crude fiber 4.i : zinc 65 ppm total ash 5.5 : cobalt 0.5 ppm calcium L phosphorus 0.95 : 0.66 : chromium selenium 1.0 ppm 0.3 ppm f. . magnesium potassium o.i6 : 0.73 ; iodine 0.9 ppm sodium 0.32 : X* iI vitamin A vitamin D , _ vitamin E ; carotene vitamin i i t i l. 8000 -15 56 12.5 c2 I.U./kg I.U./kg ftg/kg mg/kg mg/kg vitamin B, 1 vitamin vitamin B6, niacin pantothenic acid folic acid choline biotin vitamin B)2 7 5 12 55 15 4 1600 0.21 26.8 mg/kg mg/kg mg/kg mg/kg mg/kg mg/kg mg/kg mg/kg ug/kg g/100 g g/16 g N g/100 g g/16 g isoleucine 0.79 4.0 valine 1.00 5-1 *- leucine 1.52 7.7 arginine 1.08 5.5 lysine 0.99 5.0 hi atidine 0.43 2.2 I" i me thionine 0.40 2.0 alanine 1.02 5.2 L cystine 0.32 1.62 aspartic acid 1.65 8.4 phenylalanine 0.84 4.3 glutamic acid 3.48 17.6 rj tyrosine 0.51 2.6 glycine 1.06 5.4 U threonine 0.77 3.9 proline 1.22 6.2 tryptophan 0.22 1.11 serine 0.94 4.8 L- I) not ana,lysed. r I Crvo-TTJO, June 1982 L: i` L L T 1 L_ *, i L_ f -- ri i. i i V r [ l 1 tw : L r | i ( L L 'r. L (' G L ANNEX 8 - 79 * CONTAMIHANTS IN BASAL DIET FOR RATS AKD MICE Sample of a 1000 leg batch produced on January 24, 1980 lead cadmium mercury tin arsenic selenium organo-P- compound s organo-Cl-compounds 0.95 o.oe 0.02 2 0.4 0.24 ng/kg ag/kg ng/kg vg/kg ag/kg ag/kg HD HD dlthiocarbamates (as CS^) aflatoxin Bj'B2'Gl'G2 oestrogenic activity (Tiecco test) 0.5 tg/kq < 5 uq/kq HD beniofa)pyrene benzo(b)fluoranthene indeno (l,2,3-c,d) pyrene benzo(k)fluoranthene fluoranthene K-nitrate Na-nitrite nitrosodimethylamine nitroeopyrrolidine other nitrosamines urease activity trypsin inhibitor haemagglutinins positive dilution 2.5 4.4 2.8 2.6 12.0 ug/kg ug/kg ug/kg ug/kg ug/kg 163 g/kg 1 mg/kg < 0.4 <2 HD 0.15 < 0.6 none ug/kg ug/kg 6pH mg tryps/g sample ND - not detectable CIVO-TNO-May, 1980 -- - j ij ! j j X 99 CO 03 pa ro 03 03 ANNEX 9 CONTAMINANTS IN BASAL DIET FOR RATS (1000 kg btch produced 27 Feoruary 1981) lead cadmium mercury c in arsenic selenium 0.3 0.07 0.25 < 1.0 0.35 0.4 mg/kg mg/kg mg/kg mg/kg mg/kg mg/kg organo-Pwcompounds organc-Cl-coopounds PCB * t dithiocarbamates ND N0 ND ND 80 R&S 134239 aflacoxin B2.Gj_.C2 oestrogenic activity (Tiecco test) benzo(a)pyrene benzoC b)fluoranthene indeno (l,Z,3-c,d) pyrene bento(k)fluoranthene fluoranthene K-nitrate Na-nitrite nitrosodimethylamine ni troaopyrrolidine other nitroaaoinea urease activity trypsin inhibitor haeaagglutinins positive dilution negative dilution <5 P g/kg ND 0.8 0.9 0.6 0.3 5.7 u g/kg p g/kg p g/kg P g/kg P g/kg 77 mg/kg 1 mg/kg 1.2 P g/kg 1.7 P g/kg ND 0.05 A pH < 0.6 mg tryps/g sample 1 : 10 1 : 20 NO * not detectable CIVO-TNO, June 1981 T_iSr'_ L. 1 V -- V. Lt r~ i [ r1 t_- r r i L r L r r 1- f i L. h L I t f L t1 V.H'bA V CONTAMINANTS IN BASAL DIETS FOR RATS (1000 kg batch, produced 9 September 1980) lfi4d cadmium mercury tin arsenic selenium organo~P_compounds organo~Ci-compounds 0.25 mg/kg 0.045 mg/kg 0.025 mg/kg < 1.0 mg/kg 0.45 mg/kg 0.40 mg/kg ND ND dithiocarbamates ND aflatoxin BpBj.Cj^ and Cj oestrogenic activity (Tiecco test) <5 Ug/kg ND benzo(a)pyrene benzo(b)fluoranthene indeno (l,2,3-c,d) pyrene benio(k)fluoranchene fluoranthene 2.0 2.7 2.0 1.0 16.0 Ug/kg U g/kg ug/kg Ug/kg U g/kg K-nitrate Na-nitrite 80 mg/kg l mg/kg nitroaodimethylamine nitroaopyrrolidine other nitroaamine* 0.5 1.5 ND U g/kg U g/kg urease activity trypiin inhibitor haemagglutinins positive dilution negative dilution 0.05 <0.6 none - A pH mg tryps/g sample ND - not detectabli CIVO-TNO deceober 1980 * 1 ' | i 3J 0 /) pJt CO K to O / ANNEX I I CONTAMINANTS IN CIVQ BASAL DIET FOR RATS (4000 leg batch, produced by Hope Farms on September 9, 1981). 1 ead cadmium mercury t in arsenic selenium 0.9 mg/kg 0.07 mg/kg 0.005 mg/kg < 1.0 mg/kg 0.45 mg/kg 0.25 g/kg organo-P-compounds me 1 a th i on organo-Cl-compounds PCB's d ithiocarberaetes (as CS-,) ND 0.08 ND ND < 0.5 mg/kg mg/kg af1ttoxin bi,B2,C1pC2 oestrogenic activity (Tiecco test) <5 u g/kg ND bemo(a)pyrene benio(b)fluoranthene indeno (l,2,3-c,d) pyrene benzo(k)fluoranthene fluoranthene bent(ghi) perylene 0.4 0.7 < 0.5 0.1 0.9 0.3 ug/kg ug/kg ug/kg ug/kg ug/kg V g/kg K-nitrate Na-nitrite 83 <1 mg/kg mg /kg nitrosodimethy 1 amine nitroaopyrrolidine other nitrosamines urease activity trypsin inhibitor haemagglutinin* positive dilution negative dilution 0.9 1.0 ND u g/kg U g/kg 0.05 A pH 0.51 mg trypi 1 : 50 1 i 100 R&S 134241 ND not detectable CIVO-TNO. November 1981 ANNEX 12 CONTAMINANTS in crvo BASAL diet for rats (A000 kg batch, produced by van Eck, Cothen, on Febr. 9, 1982) Lead cadmium mercury C in arsenic selenium 1.4 0.09 0.003 < 1.0 0.3 0.3 mg/kg mg/kg mg/kg mg/kg mg/kg mg/kg organo-P-compounds organo~Cl-compounds PCB's dithicarbamates ND ND ND < 0.5 mg/kg aflatoxin B..B .G.,G^ 1 21 2 oestrogenic activity (Tiecco test) <5 ND Pg/kg benzo(a)pyrene 0.5 benzo(b)fluoranthene 0.1 indeno (1 f 2,3-e , d) pyrene < 0.1 benzo(k)fluoranthene < 0.2 fluoranthene 0.2 benr(ghi) perylene < 0.3 K-nitrace Na-nitrite 34 ND pg/kg Pg/kg Pg/kg Pg/kg Pg/kg Pg/kg mg/kg nitroaodime thylamine nitroaopyrrolidine o^her nitrosaminea NO 0.7 Pg/kg ND urease activity trypsin inhibitor haenagglutinins 0.00 4 pH 1.4 mg tryp*/g sample positive dilution none negative dilution ND " not detectable CIVO-TNO, June 1982 - 3 - R&S 134242 * i t. i j t1 r j i V. r 1 L r 3J (/> 03 l 4* ro CO f i r ' \ A-WlX 13 Contaminants In drlnhlnc water contaminant - lnorcar.ic substances (yg/1) arsenic cadraiurn chromium copper cyanide lead mercury selenium cine potassium nitrate sodium nitrite ammonia - organochlorine confounds tug/1^ 1,1-dichloroethane dichloroaethane tetrachloroethene tetrachloronethane 1,1,1-trichloroethane trichloroethene trichloroaethane HCB lindane (y-HCH) o-BCH 6-HCH heptachlor heptachlorepoxide c-chlordone y-chlordane aldrin dieldrin endrln methoxychlor p,p'-DDE p,p'-TDE 0,p`-DOT p,p*-DDT PCBe - organophosphorus compounds (ug/1) chlorpyrlfos dlatinon dichlorvos fenitrothlon malathion mevinfo* parathion sulfotep detection lir.lt 10 1 10 1 100 1 1 1 1 1,000 1,000 500 6 tentative fcaxir.ur'. so 5 50 50 100 SO 1 10 100 50,000 1,000 1,000 1 5 1 0.1 0.1 0.5 0.3 0.01 0.01 0.01 0.03 0.01 0.02 0.03 0.03 0.02 0.03 0.05 0.15 0.03 o.oe 0.06 0.1 0.8 0.1 0.1 0.1 0.2 0.2 0.1 0.2 0.1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 0.1 0t 1 0.1 0.1 0.1 0.1 0.1 0.1 L L ANNEX 13 (CONT.) Contaminants m drinklnc water (cont.) contaminant detection limit - tentative maximum gol^cUc aromatic hvdrocarbons (ng/l) benio tb)fluoranthene bento(k) fluoranthene bento(ghi)perylene bento(a)pyrene i fluoranthene indenod ,2,3-cd)pyrene S 200 5 200 5 200 5 200 10 200 5 200 R&S 134244 r ! 1. i. I Ur- L i u l Nov. 1979 L L O O is R&S 134245 ANNEX I4 86 CONTAMINANTS IN TAP WATER (determined in period Nov.-Dec, 1979) lead cadmium mercury arsenic elenitra copper chrocd.ua zinc cyanide Ob <1 *1 5 2 *1 SO <1 5 3 arnnonla nitrate nitrite chloride sodlua C&lclum potassium iron EEE 9.6 < 0.01 23 14. S 31 1.2 0.04 PPfa dlchlormaethano <S BCS 1,l*-dichloroethane chloroform absent 0.8 O-BCB lindane 0 1,1,1-trichloroethane 0.2 heptachlor totrachloroae thane 0.5 heptachl, epoxide trichloroethane te trachloroethene absent absent E21 aldrin dieldrln a-chlordane benzo(a)pyrene <S T-chlordane benzo(b)fluoranthene <5 endxin indeno (l,2,3,-c,d) pyrene* 5 p,p'-DDE benzo (k) fluoranthene <S o,p'-DDT fluoranthene < 10 p,p'-Tt>E benzo (g,h,i) perylone < 5 p,p'-DDT fl-HCB PCS'# DDVP mevinfoe diazinon malathlon parathion chlorpyrifos sulfotep ienitrothion EES. < 0.01 < 0.01 < 0.01 < 0.01 < 0.02 < 0.02 < 0.03 ' < 0.03 < 0.03 < o.os < 0.03 < 0.06 < 0.06 < 0.10 < 0.03 < 0.8 < 0.1 < 0.1 < 0. 1 < 0.2 < 0.2 < 0. 1 < 0.1 < 0.2 ANNEX 15 67 ; ON TAMINANTS IN TA? WATER (determined in period Jan.-April 191 c ad :admiuro wrcury irsenic Jeleniin -'tipper rhromiun :inc ryanide b ; < 0. ] 0. 1 < i 5; *3 3 <2 anroonia nitrate nitrite chloride sodium calciua potassium iron PP 0.01 11 < 0.01 30 18.5 31.0 1.3 0.06 R&S 134246 EL iichloromethane i,1-dichloroethane <h <1 ;hlorof orro 0, 3 I,1,J-trichloroethane < 0,3 :etrachloromethane < 0, 3 irichloroethene 0.3 :etrachloroethene < 0.2 EE! >enzo(a)pyrene >enzo (b) fluoranthene < => *5 .ndeno (1,2,3,-c.d) pyrene< 5 jenzo (X) fluoranthene < fluoranthene < 10 (g.h.i) perylene <S HCB a-HCK lindane heptachlor heptachl* epoxide aldrin dieldrin o-chlordane Y-chlordane endrin p,p'-DOE o, p'-DDT p , p'-TDE , p * -DDT P- ''CH PCB 1 DDVP roevi nfos diazinon tnalathion parathion chlorpyrifos sulfotep fenitrothion EE* < 0.01 < 0.01 < 0.01 < 0.01 < 0.02 < 0.02 < 0.03 < 0.03 < 0.03 < 0,05 < 0.03 < 0.06 < 0.06 < 0.10 < 0.03 < 0.8 < 0.1 < 0. 1 < 0.1 < 0.2 < . 0.2 < 0.1 < 0.1 < n7 ANNEX 16 CONTAMINANTS IN TAP WATER (determined in period Sept.- Nov. 1980) -8 R&S 134247 lead cadmium mercury arsenic selenium copper cb romium cine cyanide E^ <1 < 0.1 0.1 1 <1 50 <1 10 <1 ammonia nitrate nitrite chloride eodiuo calcium potaaaium iron dichloromethane 1p1--dichloroethane chloroform 111,1-trichloroethane tetrachlorooethane trichloroethene te trachloroethene ^ <5 0.3 0.3 0.1 < 0.1 0.4 < 0.2 benro(a)pyrene <5 benro(b)fluoranthene <5 indeno (1,2,3 ,-c(d)pyrene < 5 benro(k)fluoranthene <5 fluoranthene < 10 benro(g,h,i)perylene <5 HCB o-HCH 1 indent heptachlor heptachl. epoxide a ldrin d ieldrin a -chlordane T -chlordane endrin p.p'-DDE o, p'-DDT p.p'-TDE p.p'-DDT 6-HCH methoxychlor ppm < 0.01 8.0 < 0.01 25 15.0 28.5 1.2 0.01 pb < 0.01 < 0.01 < 0.01 < 0.01 < 0.02 < 0.02 < 0.03 < 0.03 < 0.03 < 0.05 < 0.03 < 0.06 < 0.06 < 0.1 < 0.03 < 0.15 PCB'a < 0.8 CIVO-TNO December, 1980 DDVP mevinfoa diarinon malathion parathion chlorpyrifoa sulfotep fenitrothion < 0.1 < 0.1 < 0.1 < 0.2 < 0.2 < 0.1 * 0.1 < 0.2 ANNEX 17 CONTAMINANTS IN TAP WATER (determined in period March - May 19B1) lead cadmium mercury arsenic selenium copper ch romiuxn zinc cyanide b 1 < 0.1 < 0.1 2 <1 30 <1 2 6 amraonia q!trace nicrite chloride sodium calcium potassium iron m < 0.01 16 < 0.01 25 14.0 30.0 1.1 0.03 dichiorooethane 1,]~dichloroeLh*ne chloroform 1,1,1-trichloroethane tetrachloromcthane tricxxloroeLhene Pb <5 <1 < 0.1 < 0.1 < 0.03 1.0 tetrachloroethene beozoC *)pyrene E <3 benzo(b)fluoranthene <3 indeno {1,2,3t-c,d)pyrene < 5 benzo(k)fluoranthene <3 fluoranthene <10 benio(g,h','i)pery lene <5 HCB a-HCH lindane heptachlor heptachl* epoxide al*drin dieldrin o-chlordane X-chlordane endrin p,p1-DDE o.p'-DDT p.p'-TDE p.p'-DOT 6-HCH oethoxychlor PPb < 0.01 < 0.01 < 0.01 < 0.01 < 0.02 < 0.02 < 0.03 < 0.03 < 0.03 1 0.05 < 0.03 < 0.06 < 0.06 < 0.1 < 0.03 <0.15 R&S 134248 O V PCB'a < 0.8 CIVO-TNO May 1981 DDVP meviofoa diatinon malethion parathion chlorpyrifoa aulfotep fenitrothion < 0.1 < 0.1 < 0.1 < 0.2 < 0.2 < 0.1 < 0.1 < 0.2 i /% o V R&S 134249 ANNEX 16 CONTAMINANTS in CIVO TAP WATER (determined in period Sept. - Nov. 1981) - 90 - lead cadmium mercury arsenic selenium copper chromium zinc cyanide ob <1 <1 <S < 10 <1 50 < 10 5 < 10 dich1orome thane 1.1 -dichloroethane chloroform 1,1,1-trichloroethane te tra ch1orome thane trich loro e the ne tetrachloroe thene b <5 <1 < 0.1 0.2 0.2 1.5 benzo( a) pyrene 221 <2 benzo(b)fluoranthene <2 indeno (1,2,3,-c,d)pyrene <10 benzoC k)fluoranthene <2 fluoranthene <1 benzoCg ,h , i)pery lene <3 mmonia nitrace nitrite chioride aodium calcium potassium iron < 0.01 9.3 < 0.01 30 17.5 30.0 1.2 0.03 HCB a-HCH lindane heptachlor heptachl. epoxide aldrin dieldrin o-chlordane y-chlordane endriri p, p *-DDE o, p'-DDT p.p'-TDE p.p'-DDT 6-HCH me thoxychi or Db < 0.01 < 0.01 < 0.01 < 0.01 < 0.02 < 0.02 < 0.03 < 0.03 < 0.03 < 0.05 < 0.03 < 0.06 < 0.06 < 0.1 < 0.03 < 0.15 PCB's < 0.8 CIVO-TNO May 1982 DDVP mevinfoa d iazinon ma lath ion para chion chlorpyrifoa ulfotep fenitrothion < 0.1 < 0.1 < 0.1 < 0.2 < 0.2 < 0.1 < 0.1 < 0.2 ANNEX 19 CONTAMINANTS IN TAP WATER (determined in period Jan - March 1982) R&S 134250 1 ead cadmium mercury arsenic selenium copper ch romium zinc cyanide b <1 <1 <5 < 10 <1 AO < 10 10 < 10 dich1orome thane 1, l-'dichloroethane chiorofoim 1,1,1-trichloroethane Pb <5 <5 < 0.5 < 0.2 te era chi orotne thane trichloroethene te trachloroe thene < 0.1 0.4 < 0.2 PPC benro(a)pyrene < 1.3 benzo(b)fluoranthene < 0.5 indeno (1,2,3-c,d)pyrene < 1.3 benzo(k)fluoranthene < 0.7 fluoranthene benzoCg ,h,i)perylene < 0.5 < 5.4 atmonia nicrate nitrite chloride sodium calcium potassium iron =" < 0.01 6.3 < 0-01 23 13.0 30.0 1.1 0.02 KCB a-HCH 1indane heptachlor heptachl. epoxide aldrin dieldrin o-chlordane y -chlordane endrin p.p'-ODE o.p'-DDT p.p'-TDE P.P'"DDT 6 "HCH me thoxyehlor ppb < 0.01 < 0.01 < 0.01 < 0.01 < 0.02 < 0.02 < 0.03 < 0.03 < 0.03 < 0.05 < 0.03 < 0.06 < 0.06 < 0.1 < 0.03 < 0.15 PCB's < 0.8 Civo-TNO May 1982 DDVp mevinfos diatinon malathion para thion chlorpyrifos sulfotep enitrothion < 0.1 < 0.1 < 0.1 < 0.2 < 0.2 < 0.1 < 0.1 < 0.2 r i 3D B CO _i co ro i cn '' - 1 j r L- ii l| j L \ I L r| L r L r 1 U r L P 1L_ r r L, - ANNEX 20 j/INCj - *. OSS *EF ERENCE L IST ING AY NO. f. n 'ERR RHF kk'FNLt HAlt - pi, i-: MAI t S FFI4ALkS OMR 0002k 0004K '00060 0008N '003 00 '0012k '00140 '00160 '001BI '00200 '00220 '00240 '00260 '002B0 '00300 '00320 '00340 '0036N '00380 '00400 '00421 '00440 '00460 '004 BO '00500 700520 '00540 'O0560 '00580 '00600 '00620 '00640 70066N 700680 '00700 700720 700740 70076N 70Q78K 700800 700B20 700840 700860 700880 700900 700920 700940 70096N 700y8Li EX >"R A `l ft A ft 6 ft 8 A 30 A 12 ft 14 A 16 A 18 A 20 A A 24 A 26 A 28 A 30 A 32 A 34 A 36 A 38 A 40 A 42 A AA A 46 A 48 A 50 A 52 A j4 A 56 A 38 A 60 A 62 A 64 A 66 A 48 A 70 A 7? A 74 A 76 A 78 A HO A 82 A 84 A 86 A 88 A VO A 92 A 94 A 96 A V8 8IJR (0) (0) (0) (0) (03 (0) (0) (0) <03 <0 3 (03 (03 (0) (0) <03 (03 <03 (03 ((3 3 (0) (0) (0) (03 (0 3 (0) (0) <03 (03 (03 (0) (0) (03 <0> 0) v(l) (03 <03 <03 (0) (03 (03 (03 (0! <03 (0) <03 <03 <03 <03 EAr.h n p J. \' 4 /I A A A 6 6 6 6 6 8 8 8 8 8 10 30 10 10 10 12 1 : 12 12 12 1A i4 .1 A 14 34 16 16 16 16 36 18 18 1H 18 18 20 :*o 20 JO EAKR 7 R1 K2 LI L2 R1L1 R31.2 K2I. 3 R2L2 R3 7 kl K7 Ll L2 R1L1 ML 2 K2I. 1 K2l2 R3 7 kl K2 Ll L2 R1L1 kill' R2I. 1 k2L2 R3 7 kl K2 Ll L2 R1L 3 R1L7 R2I. 3 R2L2 R3 7 Kl R2 Ll L2 R1L 3 R11.2 R2L 1 R2L2 COrtR 7700018 7/0003)1 7700058 7700078 7700098 77003 3 k 770013)i 7700158 770017k 7700198 7700235 77002311 770025k 7700278 770029D 7700338 770033k 7700358 770037k 7700398 770043 K 770043)1 770045k 770047k 7700498 770051k 770053)i 7700558 7700578 7700595 770063 8 7700631 770065k 7700671 7700695 7700718 7700735 7700755 7700775 7700791 7700815 7700H3K 7700855 7700875 7700898 7700918 7700938 7700958 7700978 kXPK A3 A3 A5 ay AV A 11 A 13 A 15 A 37 A 19 A 21 A 23 A 25 A 27 A L'V A 31 A 33 A 35 A 37 A 3V A 41 A 43 A Au A 47 A 4V A 51 A 53 A 55 A 57 A 59 A 61 A 63 A 65 A 67 A 6V .i 71 A 73 A 75 A 77 A 79 A HI A 83 A 85 A 87 A H9 A 91 A 93 A 95 A 97 ink cahh: ARK (0) <0> (0) <()) <03 (0) (0 ) (0) (0) (0) (0) (0) (0) (0) (0) (0) (0) (0) <<>> U) ) (0) (0) (0> (0) (0) (0) (0) (03 (0) (0) (0) (03 (03 (0) <03 (0) (0 3 (0) (0) (0) (0) (0) (0) (0) (0) <03 (0) (0) (0) it ik i I;.1 i Ll L2 2 R1L 1 3 KILL 3 K2\. 1 3 K2L2 '/ R3 T 7 . k i K3 ,J l 1 fTJ 12 7 R1L 3 7 Kll.2 / k:.'i. i 7 K2L2 7 R3 9 7. 9 R1 9 k:? 9 Ll 9 L2 11 R1L 3 1 1 K1L7 1 I .1 11 K2L2 1 1 R3 13 7. 1 3 k.( 13 k*:,1 13 Ll 13 l 2 15 R1LJ 15 k 31. 2 15 R2I. l 15 R2L2 15 R3 17 7 17 Kl 17 R'J 17 Ll 17 L 2 19 R3L 3 19 K.U 19 k:*l. 3 iy K2L2 K - killed D * dead I - ill - (dead) - % .j# -- 1 1 ! i j L w / riL _ R&S 134252 ii j L r~ il j i [ L I L i-A L r- L. -- A-WEX 20 (C0N7. 1) 1/ i Nil 93 OSS REF E R E N C E L 1 STING .-,1 NU. 125 0 ZhklJ k`K F 1 KF Nl f DA I L - 21 hALES KFFI ALES rik oioori '010211 '01011' ' 010 6 D '010131' '01 1 0L< '01120 'Oil -in '01160 '01180 701200 701220 701210 701260 /01280 701300 701320 701310 701360 70138K 7011OK 701120 701110 701160 701180 701500 701520 701511 701560 701580 701600 70162K 70161K 70166K '7016BK '70170K '701720 '701710 701760 '70178K '70180K '701820 '701810 7701860 7701880 770190K 7701920 770191K 7701960 7701980 7702000 h XPk A100 A102 A1 01 A1 06 A1 06 A110 A 112 A1 1 1 A1 1 6 A1 18 A1 20 A122 A1 21 A126 AJ 28 A130 A132 A131 A136 A138 A110 A 112 A111 A116 A11B A150 A152 A3 51 A1 56 A158 A160 A162 A1 61 A166 A168 A1 70 A172 A171 A176 A178 A180 A182 AIM A1B6 A188 A190 A192 A1 91 A1 96 A196 A200 SCR CAGE 10) 20 10) 10) 2'n7 (0) 22 (0> (0) 'ft (0) 21 (0) 21 (0) 21 (0) 21 (0) 71 <0 ) 26 (0) 26 (0) 26 <0> 26 <0> 76 (0) 28 (0) 28 (0) 28 (0) 28 <0) 26 (0) 30 (0) 30 (0) 30 (0) 31) (0) 30 10) 32 (0) 32 (0) 32 (0) 32 (0) 32 (0) 31 (0) 31 (0) 31 (0) 31 ( 31 ( It 1 36 <0> 36 (0) 36 (0) 36 (0) 36 (0) 38 (0) 38 (0) 38 (0) 36 (0) 3H 10) 10 to ) 10 10) 10 (0) 10 10) 10 EARR K3 l R1 k? l.l 1.2 R1L1 R1L2 R71 1 R2L.2 R3 7. R1 K? 1. 1 1.2 R1L1 R1L2 R21.1 R2L 2 R3 Z R1 R2 1. 1 1.7 R1L1 R1L2 R2L 1 K21.7 R3 Z R1 K? 1.1 L2 R1L1 R1L2 R2I 1 R2L2 R3 Z R1 R2 LI L2 R1L1 R1L2 R2L 1 R2L2 R3 C.ClrtR 771)099)' 770101D 770103H 7701051' 7701071' 770109L' 7701111 7701131 770115K 77013 7)' 7'701i?H 7701211 7701231 770125K 7701271' 77012VD 7701311' 770133K 7701351' 770137D 770139U 770111K 770113P 7 7011511 7701171.' 771)1191' 770151H 770153H 7703 551.1 770157)' 770159K 7 7 0161D 770163H 7701 651) 770167K 7701691' 7 70171D 770173K 7701751' 7701776 770179D 770181D 7701831' 7701851.' 770187D 770189K 770191K 7701931' 770195).' 7701971' . 770199k F XF'k SGP CAGE EARR A 99 10) ) 9 k'3 A 1 01 10) :.'i / A1 03 10) 21 ftl A 105 10) 21 R2 A1 07 ' 10) 21 1, .1 A109 (0) 2.1 i :/ A11 1 (0) 25 R j 1. 1 A11 3 1 0 ) 23 R1L2 A1 15 10) 23 R2L 1 Al 3 7 1 0) 23 k ;i All? (0) 23 R3 A121 10) 25 7 Al 23 (0) 25 R1 A i 25 (0) 25 k? A127 (0) 25 U A129 (0) 25 1.2 A1 31 10) 27 Rl LI A133 (0) 27 R1L2 A135 (0) 27 R2L1 Al 37 10) 27 k2l. 2 A139 (0) 27 R3 Al 1 1 10) 29 Z A113 10) 29 Rl A115 (0) 29 R2 A] 17 (0) 2? 1.1 Al 19 (0) 2? 1.2 Al 51 (0) 31 kILl A153 10) 31 R1L2 A) 55 (0) 31 k`2L 1 Al 57 10) 31 R2I. 2 A159 10) 31 K.i Al 61 (0) 33 Z A163 (0) 33 Rl A165 10) 33 K3 A167 10) 33 1. t Al 69 10) 33 1.2 A3 73 10) 35 KILl A173 (0) 35 R1L2 A175 10) 35 k?l.l A177 10) 35 R21. 2 Al 79 10) 35 K3 Al 81 (0) 37 1 A.163 10) 37 Rl A185 (0) 37 k2 A187 10) 37 1. 1 A189 10) 37 12 A] VI 10) 3y R1L1 Al 93 10) 39 R1L.2 A195 A197 10) 10) 39 3V R2L.1 R2L2 A jl 99 (0) 39 R3 I l l r L r \ !ri !*- i 1 ! t i ! (' t r Jj ffo C/A/j CO to CJl CO 1 f L (11 ' L L II L annex 20 (COST. 2) :> MNU - 94 ~ m r OSS REF E ft E N C IST ING A3 NO. 123 0 ZElft'U RtF L KkNt t ) Ir-I 1 r ` l* males FEMAL r. S OMR 02021' 020-10 02061' 020Si' '02100 '02120 '023 -10 '02160 '02190 '0220K '02220 702241' 70226K 702280 702300 702320 70234K 702360 702380 702-100 702-12K 702-140 70246K 702480 702500 70252K 702340 702560 7025SK 702600 '702620 '702640 702660 702680 '702700 ' 702720 ' V0274K 7 70276K -702780 7702800 7702820 7702840 7702860 7702BBI 770290K 7702920 770294K EXFR P2 04 B6 P8 P 10 P 12 P 14 P 16 P. IK P 20 P 22 p 24 P 26 P 28 P 30 h 32 P 34 P 36 P 38 P 40 P 42 B 44 P 46 P 48 P 30 P 52 B 54 B 56 H 38 B 60 P 62 B 64 P 66 H 6H P 70 P 72 P 74 P 76 P 78 P 80 B 82 P 84 P K6 B 88 P 90 B 92 P 94 SGR LARK (0) (0/ (0) (0 3 (0) (0) (0) <0 3 (0) (0) (0) (0) (0) (0) (0) (0) (0) <03 <03 (0) (0) (0) (03 (0) <03 (03 <03 (0) (03 (03 (03 (0) (03 (0 3 <03 (0) (03 <03 <03 <03 <03 <03 <03 < <3 ) <03 <03 <03 47 4.7 42 42 42 44 44 44 44 44 46 46 46 46 46 48 48 48 48 48 50 50 50 50 50 52 57 57 52 52 54 54 54 54 54 56 56 56 56 56 58 58 5H 58 58 AO AO h ARK "1 Ri K2. LI L2 ftl L.l km.:; K2L1 K2L2 ft 3 7 Ri R2 L1 L2 RU.l KM, 2 R21 1 R2L2 R3 Z ft 1 K2 L1 1.2 K1L1 ftll. 2 R2L1 R2L2 R3 1 kl K2 l3 1.2 ftll. J ftll. 7 R2L 1 R2L 2 ft 3 2 Rl R2 lJ L2 K1L1 ftll 2 CUMft 77020IK 770203K 770205P 770207ft 770209ft 77021 1ft 77021.511 770215ft 770217K 770219ft 77022 iP 7702231 770225H 770227ft 7702.29P 77023 U' 770233P 770235P 770237P 770239K 7702411.1 770243B 770245B 770247K 770249P 7702515 770253ft 770255P 770257K 770259P 7702A1K 770263K 770265ft 770267K 7702AVP 77027IK 770273ft 770275K 7702771i 77027913 7702811.1 770283ft 770285ft 7702&7K 7702898 7702.9111 7702938 EXftft P1 P3 It P '7 ft 9 P 11 P 13 ft 3 5 P 17 P .19 P 21 P 23 P 25 P 27 P 29 P 31 P 33 ft 35 P 37 P 39 P 4] P 43 P 45 P 47 P 4V B 51 P S3 P 55 P 57 ft. MV P 61 B 63 P P 67 P 69 P 71 P 73 P 75 P 77 P 79 B 81 B 83 P 85 B 87 P 89 P 91 ft 93 551' LAljl F ARK <03 4 1 7 (C'3 4 i K .* (03 41 . K2 < 0 3 43 LI (0) 41 L2 !0> 43 Klt.l (0) 4 3 ft <1.2 <03 4 3 K2I 1 <0 3 4 3 R2L2 <03 43 ft 3 (03 4 5 2 <03 4 5 K.l <03 45 ft 2 <03 45 LI <03 45 L2 <03 47 ML 1 <03 4/ ft 11.2 (0) 4 / R2L1 <0 3 47 R2L? (0) 47 ft 3 <03 49 1 <03 4 V ' ft j (0) 4y R2 (03 49 L1 <03 49 L2 (03 51 Kll 1 < 0 3 5i Kll 2 (0 3 51 K2L1 <0 3 51 R2L2 <03 51 K3 <0> Li 3 7 <03 53 ftl <03 53 R2 <03 53 L1 < 0 3 53 L2 <03 5 Li ftll 1 <0 3 55 ft 11.2 <03 55 R2L1 <03 55 R2L2 (03 55 K3 <03 57 7 <03 57 ftl (03 57 ft 2 (0) 57 l1 (0) 57 1. 2 <03 5 V Kll 1 (03 59 R 11 2 ANNEX 20 (COST. 3) 71 NO OSS REFERENCE LI STING Y NO. 125 (i MALES 3MR tXKK 32961' 22980 03008 03020 03040 03060 0308K 03100 03120 031 4 0 03161 03180 03200 03220 03240 03260 '03280 '03300 '03320 '03340 '03360 '03380 '03401 703420 '03440 703460 703480 70350K 703520 70354K 703560 7035B0 70360K 703620 70364K 703660 703680 703700 '70372K '703740 '703760 '70378K ' 70380K '703B20 '703840 7703860 7703880 7703901 7703920 7703940 7703960 770398K 7704000 0 96 0 98 0100 810? HI 04 0106 0108 0110 03 12 0114 0116 0118 0120 0122 0124 0126 0128 0130 0137 0134 0136 0138 0140 03 47 0144 0146 0148 0150 0 3 57 0154 0156 0158 0160 0 3 67 0164 0166 0168 0170 0 3 72 0174 0176 0178 03 80 01B7. 0184 0186 0188 03 90 0 3 V2 0194 03 96 0198 0700 5CCR l-.nGC EARR CO) cO) CO) <C>) CO) CO) CO) CO) ( 0> (0) CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) (0) CO) CO) CO) CO) (0) CO) (0) <0i CO) C0> CO) CO ) CO) CO) CO) CO) ( 0) CO) (0) <0 3 CO) CO) CO) CO) CO) (0) CCD CO) CO ) CO) CO) 60 K7L 3 60 K7I 2 7.0 K3 67 Z 62 k ] 62 K2 62 LI 7,2 L 7 64 F: 1L1 64 R1L2 64 R7I 1 64 K7I 2 64 K3 66 2 66 Kl 66 K7 66 1.1 67. 1.2 68 R1L1 68 ft) t 2 68 K2I. 1 68 K7I.2 68 R3 70 Z 70 KJ 70 K? 70 1 1 70 L2 72 R1L1 77 K 3 L 7 77 ft 71. ). 77 ft7! 2 77 ft 3 74 Z 74 ftl 74 k:.' 74 i i 74 L2 76 R1L 1 7<s ft 31 2 76 ft 21 1 76 K7L 7 76 R3 78 Z 7H ft) 7H K2 78 1 3 78 L2 BO Kl L 1 130 KIL2 80 ft2L 1 BO K2L7 BO K3 Zl. Nil ft I lll:l N l 1 L V h MAi F S COMR h Xftft Hftft (.AGE EAKK 770295 ft 0 95 CO) 59 K2L.3 7702978 o y" cO) 5V K2I ' 7702998 8 99 (CD 59 ft.) 7703018 03 01 CCD 6) 7703031 0)03 CCD 61 2 Kl 7703050 0105 CO) 61 K.' 7703078 0107 <03 61 1. 1 770309K 8109 CO) A) 1. 2 7703118 03 11 CCD 63 FC1L1 770313) 8113 CO) 63 K.1.L2 7703158 0115 CO) 63 K2l 3 7703178 B117 CO) 63 K3C.2 7703198 8119 CCD 63 K3 7703218 0 3 2 3 CO) 61. 7. 7703231 0123 (0) 65 ft 3 7703258 8125 CO) 65 K.' 770327K 0127 <03 65 1. 1 7703298 8129 (CD 65 L2 7703318 0131 CO) 67 R1L3 7703338 8133 (0) 67 KILL 7703358 8)35 CO) 67 KOI 7703378 0137 (03 67 K2i. : 7703391 0139 CO) 67 ft 3 7703418 8141 (0) 69 3 7703438 8143 (0> 69 R3 7703458 0145 CO) 69 ft 2 770347K 8147 (0) 69 1. i 7703498 0149 CO) 6 V L2 770351K 0151 (0) 71 R1L3 7703530 8153 (0) 71 Kl L 2 7703558 0155 (0) 71 K2| 7703578 0157 CO) 71 K3I.2 7703598 0359 an 71 ft 2 7703618 0161 .0) 73 Z 770363) 0163 C0> 73 R1 7703650 0165 CO) 73 K2 7703678 0167 (0) 73 11 770369K 01 69 CCD 73 L2 7703738 0171 CO) 75 R1L1 7703738 8173 (0) 75 RH .' 7703758 0175 (0) 75 K2I 7703778 0177 (0) 75 K2I. 2 7703798 0179 (0) 75 ft 3 7703B3 8 OlBl (0) 77 7703H38 0183 (0) 77 7 k< 7V03B38 B1B5 CO? 77 ft ' 7703B7K 0187 CO) 77 11 770389K 03 H9 (0 3 77 L7 77039) ti 0 > 9 3. CCD 79 R3 L 1 7703938 81 93 CCD 79 ft 3 C . 7703958 0195 CCD 79 R2I 1 7703978 0 ! 9/ (0) 79 K21 2 77C)39y;i. 0)99 (0) 79 ft.-- R&S 134254 - r k 1 * ; i 30 <Z> -a CO fe ro CJl CJl A57CEX 20 (CONT. 4) 1/~l NU - 96 - OSS R E F E R E N C E L I STING it NO. 175 0 71 KU RFF ^ kf- Mi 1 ) i; 1 1 y * HOLES FEhOLtS L'MR 0 ^ 0 "> LI Q404k 04060 04osn cm] on '0412K '0414k Q416K 04 1BK 04 200 '04220 *04240 '04260 '04230 *04300 *04320 *04340 '04360 *04380 /0440D /0442D 704440 704460 704480 /0450K 704520 704540 704560 704580 /04600 70462K 704640 70466K 70468K 704 70K 704720 70474K 704760 704780 704800 704820 704840 '704860 '704880 704900 EXF'R c C C: 6 i; H c: 10 C 12 C 14 t. 16 ( 18 n 20 c *> -> C 24 t; '.'6 <: 28 8 30 c T 'i L 34 i; 36 L 38 c 40 c 42 L 44 l. 'lA (. 48 c 50 c c 54 t; 56 c 58 c 60 c 62 1. (; AA i: 68 c 70 c 72 (. 74 i; /ft r. 78 c 80 c 87 8 H4 C 86 r; 88 c 90 SGF' C.OGh 1- AKK (0) w:.* 7 < 0 ) 07 R1 (0) 82 R2 (0) 82 L3 (0) 87 L7 (0) 84 kli. 1 (0) 04 K1L2 (0) 84 R2L 1 (0) 84 R2L7 (0) frM R3 (0) 86 2 <<)) 86 R1 (0) 86 R2 (0) 86 L1 (0) 86 1.7 (0) 88 till. 1 (0) BH K1L2 (0) 88 R2L1 (0) 88 R2L? (0) 88 K3 (0) 90 Z (0) 90 R1 (0) 90 R7 (0) 90 11 (0) 90 1.7 (0) V 2 R.U 1 (0) 97 R1L2 (0) 92 R2L1 (0) ?2 R2L.2 (0) 97 KA (0) S'4 7 (0) 94 CO) 94 R) R7 (0) 94 L1 <0> y^ 1.2 (0) V A kH 1 (0) 96 R1L2 (0) 96 R2L1 (0) 96 Ki'i. (0) V6 R3 CO) 90 7 CO) 98 R1 (0) 98 R2 CO) 98 11 CO) VH 1.7 LUMK 77040IK 7704030 7704050 7704070 77040VK 7704110 7704130 770415K 7704]70 7704 IS'O 7704310 7704230 7704251 7704770 7704290 770431K .7704330 770435) 770437K 77043VK 7704410 7704430 7704450 7704470 770449K 7704510 7704530 7704550 7704570 7704590 770461K 770463K 7704651 /70467K 7704690 7704710 7704730 770475K 77047/J.i 7704790 770481D 7704830 7704H5K 7704870 7/04890 LXRR C1 i3 . 5 c/ 9 c 11 c 13 t: .1 -/ i; ,1/ n ]9 c 21 L 23 i; 25 i. \yy c 29 c 31 c 33 c 35 i: 37 r. 39 c 41 c 43 i: 4 5 c 47 C 49 c 51 i. 53 i; 55 c; 57 c 59 c 6) c. 63 c Ail c 67 c 69 c 71 c; 73 <: 75 i; 77 c 79 c 81 c; 83 c 85 i: 87 c 89 SGk CO) < (i (0) CO) vO) CO) CO) CO > CO) CO) CO) (0) (0) CO) CO) (0) CO) (0) (0) (0) CO) co> CO) CO) C0) (0) CO) CO) CO) CO) CO) CO) CO) CO) CO) (0) CO) CO) CO) CO) CO) CO) CO) CO) CO) L.AGh ri i, H .1 81 81 81 Hi H.i 83 83 83 HI-. 85 85 85 85 8/ 8/ 87 B7 H7 mV 89 89 89 89 94 yi 91 91 9) ya 93 93 93 93 y:> V5 95 95 y^i 9/ 97 97 97 97 h nkk / R1 k? L1 ! !' 1; a l. i, K1 L2 R2L1 R2L 7 kV, / R1 R7 11 1./ K11. 1 R1L2 R7L 1 R7L2 K3 7. Rl R7 1. l i. :* K11 1 R1L2 R2L 1 K2I. 7 K3 / HI R? 1. 1 1. 7 K1L1 R1L 2 K2I 1 K7I. 7 K-3 z Rl K7 1i 1. 7 - *4 -- t 1 1 ANNEX 20 (CONI. 5) j/IMO -97- OSS REF E R E N C E L I S T I N G r" 3 Nil. 125 0 / h RlJ Khi-FRENLi. 8 A 1 E - 71/ H-- 1 MALI- S 1 K MAI. h H f f OUR" FXF'R SCR CAhF EARR COMR LX PR Clift 1, AfiF EARR t ! 0492D (." 92 (0) 100 Rll 1 770491 Ji C 91 (0) 99 ft") LI 04940 C 94 (0) 100 Kll. 7 770'iv 38 L 93 (0) 99 K 11 7 r 0496R C 96 10) 10(1 R7I. 1 770495K C 95 CO) 9V R71.1 04981 C 98 (0) a 00 K2L2 770497ft C 97 CO) vs- R7L2 L Q500K 05021 Cl 00 G102 (0) 100 (0) 102 R3 770499ft C VS' CO) 99 7 7705018 ClOt CO) 101 R3 7 0504 0 1:104 (0) 102 k1 770503ft Cl 03 (0) 101 Ri O506K C106 CO) 1 07 K7 7705058 C105 CO) 1 0) K"7 05080 C108 (0) 107 LI 770507ft Cl 07 (0) .id l LI 05100 C110 (0) 102 L2 7705098 Cl 09 CO) 101 L2 05120 Cll? CO) 104 Rll.) 770511ft cm (0) 103 ft] LI 05140 C11 4 CO) 104 Rll. 2 7705138 n 113 CO) 103 Ril. 7 05160 C116 CO) 1 ('4 R2I. 1 7705158 C115 CO) 1 03 ft'21.1 1 05180 05200 CUB r.120 CO) 104 CO) 104 R2L2 R3 770517ft 7705198 Cll 7 Cl 1V (0) ) 03 (0) 103 R2L2 R3 05220 Cl 22 CO) 106 7. 7705718 Cl 21 (0) 105 7 r 05240 C124 (0) 106 ft i 770573K C123 (0) 105 Ri 05260 C126 CO) 106 R2 770525ft Cl 25 CO) 1 05 ft 2 05280 Cl 28 CO) 106 LI 7705278 Cl 27 CO) A 05 LI 0530K C130 (0) 106 L2 770579) Cl 29 (0) 105 L2 05320 Cl 32 (0) 108 R1 L 1 770531.8 C131 (0) 107 ft] LI "05340 C134 CO) 108 Rll. 7 7705338 C133 CO) 1 07 Ril..:/ ' "05360 C136 CO) 108 R2L1 7705358 C135 CO) 1 0 7 R2I..1 05380 0138 CO? 108 R2L2 7705378 Cl 37 (0) .107 R2L2 05400 C1 40 CO) 108 R3 7705398 C13V (0) 107 ft 3 05420 C14 2 (0) 110 7. 77054)8 Cl 41 CO) 109 "7 "0544K C1 44 CO) 1 1 0 ft.t 770543K C14 3 (0) 10V IO. I "05460 C 1 46 CO) no k? 7705451 C1 45 CO) 109 ft',' 05480 n 48 CO) .11 0 LI 7705478 C) 47 CO) 109 LI 05500 Cl 50 (0) 110 L2 7705498 r;i 4V (0) 109 L2 "0557K Cl 57 CO) 112 Rll.) 7705518 C151 CO) 111 ft.l LI ; "0554 0 C1 54 CO) 1 17 K11..7 7705538 C153 (0) 111 1; 11. 7 "05560 C156 CO) 1 .c 7 R7I 1 7705558 C155 (0) 111 K7I 1 "05580 Cl 58 CO) i 1 7 R2L2 7705578 Cl 57 (0) 111 R2L2 f "05600 Cl 60 CO) 112 R3 7705591 (.159 (0) 111 ft 3 "05678 Cl 62 CO) 1 1 4 7 7705618 Cl 61 (0) 113 7 "05640 Cl 64 CO) 1 1 4 ft? 7705638 C163 (0) 1 13 ft .1 "05660 C166 CO) .1 i 4 R2 7705658 Cl 65 (0) 15.3 R2 "05681 01 68 CO) 1 14 LI 7705678 Cl 67 CO) 113 LI f 05700 Cl 70 CO) 114 L2 7705698 Cl 69 (0) 113 L2 i "05720 C172 CO) 1 16 Rll 1 7 7057.1 8 C171 CO) 115 R.U.1 L "05740 C174 CO) 116 Rll. 7 7705738 Cl 73 CO) 1 15 Ril 2 "05760 C176 CO) 1 c 6 K'3L 1 7705758 C175 CO) ` 1 A 5 K2L1 f "057B0 Cl 78 CO) 116 R2L2 7705778 Cl 77 CO) 115 R2L7 i "05800 C180 CO) 116 R3 7705798 C179 (0) 115 R3 i '0082K C1 82 CO) 1 I 8 7 7705H18 Cl 81 (0) 117 7 "05840 C184 CO) 1 m ftl 7 705838 C183 CO) 1 1 / RI '05861 C186 CO) l aH ft 2 770585ft C185 CO) 1 i / R2 ; "05880 1.188 CO ) 1 1 8 L. 1 7705878 1,1 HV (0) 1 1 7 LI "05900 C1 90 CO) i 18 1. 2 7705898 C189 (0) 1 1 7 17 L '05920 Cl 9? CO) l 70 R1L1 770591ft Cl 91 (0) 1 i 9 RlL 1 "05940 Cl 94 CO) 120 R1L2 7705V30 c.cy.i CO) 1 19 R1L2 '05960 Cl V6 cO) 120 R?l 1 7705958 (. 195 CO ) 11 V R71 1 i` '05900 C1 98 CO) J 70 R7I 7 770597ft C197 (0) 1IV K7I 2 1 '06000 C200 C Cl) i.70 ft" 3 7705998 m 99 ( Cl > 1 19 9`J L AI16TX 20 (CONT. 6) J7 I NO OSS REFERENCE LISTING "^Y NU 4 12j 0 ZERO Ki-hl-KHU.r Ill'll) R&S 134257 nFiLES jhk t.XRR 060 CD 060`ID 0606D 0608D 0610D 061 CD 061 4D 0 616 D 0619K 0620D 0622D 0624D 0626D 0628k 0630D 0632D 0634D 0636D '0638D '0640D '0642D '0644K '0646D '0648D '0650D '0652K '0654H '0656D CQ658D '0660D '0662D >0664R /0666D 70668K 70670K 70672D /0674K 70676D 70678D 7Q680D 70682D 70684D 706861 70688D 7Q690D 70692D 70694D D )> 4 D6 ti 8 D 10 ri 1 2 D 14 D 16 l' 10 D 20 n !*> D 24 D 26 ti 28 D 30 D 32 l.i 34 D 36 D 38 D 40 li 4 2 D 44 D 46 D 48 D so 1' e, i ,, D 54 D 56 D 58 D 60 )> 62 D 64 D 66 D 68 n 70 n /2 n 74 n 76 D 78 n WO. n 87 i.i 84 D 86 D HH 90 D 92 D 94 SGf (0> (01 (0) (0) (0) (0) (0) (0) (0) (0) (01 (0) (01 (0 1 (0) <0> (0 ) (0) (0) (0) (0) (0) <0> (0) <(>> <0> i 0) CO) (0) (0) co) (0) (> (\ (0 ) (0) (0) (0) (Ol (0) (0 1 (0) (0) (0) (0) (0) (0) CAGF \ 77 122 122 i ::> a 27 174 124 124 I 74 174 176 126 126 176 126 178 128 128 128 128 130 130 130 130 130 132 132 137 1.47 137 134 134 134 134 134 136 136 3 36 136 136 138 1 38 138 138 138 140 1 40 fc AKR I Rl K2 1. 1 1.7 ML 1 R1L2 R2L1 K7I. 7 R3 2 Rl k'7 l. 1 L2 R1L1 R1L2 K2I. 1 R7I.2 R3 Z Rl K7 L1 L2 R1L1 ML? K2I, 1 R7L 7 R3 L* KI R2 11 L2 M L1 R1L.2 R21. 1 R2L7 R3 Z Rl R2 LI 1.2 Nil. 1 Rll 2 KEhALES RUHR 770601D 770603)' 7/0605D 7/06071.' 770609K 770611D 77061 3D 7706)0 D 7/06171 770619D 7706211' 7706231' 770625R 7/0627D 770629D 770631D 7706331.1 770635H 7/0637D 77063VD 7 7 0 6 4 1D 770643D 770645D 770647D 7706491' 770651P /706S3D 770655D 770657D 770659D 7706611 7/0663K '7/06651 . 70667D 0669K 770671D 7/06/3)! 7/0675D /706771 770679D 770681D 7/06H3)! //068S.D 770687D 77061)9)! 770691D 770693D 1. XRR D1 )' ,5 D D7 D0 D 11 )i 13 )' r.. D 17 D 19 D 71 )' 23 )1 70 D 77 D 39 )' 3) !' 33 )> 35 D 37 D 39 D 4I )i 4 3 )' 45 D 47 Ii 49 D 5) )! 53 I' 55 D 57 D 5V )' 61 )! 63 D 65 D 67 li 69 J! 7 1 )> /.( D 75 D 77 Ii 79 |i 8! )' 83 D 85 Ii 87 D 89 D 91 D 93 bGF- (O) '.1) c0) 40) cci) (0) i Cl ) (0) C) (0> CO) CO) CO) CO) (0) (0) CO) (0) (0) (0) CO) CO) CO) CO) CO) (0) CO) CO) (0) CO) C0) (0) (0 ) CO) CO) CO) (0) CO) (0) (0) (0) (0) (0) (0) (0) CO) (0) cage 1 7J 12 J 121 131 * 7J 1.73 123 123 1 33 .1.2.4 135 125 125 1 75 175 1 77 127 127 17/ ! 77 129 1 ?y 129 1 29 1 7V 131 131 ) 33 131 131 133 133 133 1 3.4 1 3.4 135 135 1 35 135 \ 35 137 137 137 137 137 1 3V 139 ) ARR r1 K7 11 17 R1L1 R1L 2 R7I 1 R-'l. : R3 i. Rl K7 1. 1 L2 R1L1 M L2 R7I. 1 R7I. 7 R3 Z M R7 1. 1 L2 K1L 1 Rl L7 R 71. 1 R2I. 2 R3 Z R.I R7 13 L2 1 LI KJ L7 K2I. .1 R7L 7 R3 Z KM R7 LI 17 Ml, 1 R1L2 ANNEX 20 (CON1 J/ T NtJ 7) OSS REFERENCE LI STING 99 AV NO. 100 0 21 KO K'Er LICENCE I'ATF yi/ / Mi'.L F S F F hfil 1. b OMR EXF'K 8(11 CAIJF EARR CO MR FXPR bl;.| ' LY-iHfc EARR R&S 134258 'OavoL' -O0V8P /o7oor| '0702K '0704k '0706k '0708k '071 OR /0712k '0714k 70736k 70718k 70720k 70722k 70724K 70726k 70728K 70730k 70732k 70734K 70736N 70738K /0740K 70742k 70744K 70746K 70748K 70750k 70752K 7 07lJ4K 70756k 70758k 70760k 70 7a2K 70764k 70766k 70768K 70770K 70772k '70774k '70776k '7077EJK '70780k '70702k '70784k '70786k '70788k '70790k 7707V2k V0774K /70796K 770798k 770800k )i V6 P 9ri ni oo HI 02 m 04 111 0 6 1.1108 I'llO 0112 Hi 14 Ml 1 4 rmo PI 20 L112 2 Pi 24 P176 1.1178 Pi 30 PI 32 P134 Ml 36 PJ 38 1*1 40 PI 42 Pi 44 PI 46 113 48 Pi 50 PI 52 P1 54 iMS/i p] 58 PI 60 Pi 62 PI 64 Pi 44 P) 68 Pi 70 Pi 7? p] 74 PI 76 PI 78 Pi 80 Pi 82 Pi 84 PI 86 PI 8Fi P] 90 PI 92 PI 94 Pi 96 p] 98 P'-.'OO (0) (0) (0) (0) (0) (0) (0) (0) (0) <0> (0) (0) (0) (0) (0) (0) (0) (0) (0) (0) (0) (0) <01 (0) (0) (0) <0> (0) (0) (0) (0) (0) <0) (0) (0) (0) <0> CO) (0) (Oj <01 (0) (0) (0) CO) (0) (0) (0) 10) (0) (0) (0) (0) 1 40 140 140 147 142 1 43 142 1 42 ;! M .144 1 44 144 144 .14 6 146 1 46 146 1 46 148 1 48 148 148 1 48 150 150 150 150 150 153 1 53 152 152 153 i t>4 154 154 154 154 156 1 j6 156 156 156 15 FI 158 158 15FJ 158 160 160 160 160 160 R2L1 R2L2 ft 3 2 R1 R2 LI L2 R11. 1 R1L2 R2L1 R2L2 R3 7 R1 R2 11 1.2 R.ll. 1 R1L2 R2L1 R21 2 r:\ 7 R1 R2 11 1.2 kill R1 L 2 R2L 1 K2I 3 K.1 7 R1 R2 1. 1 1. 1* K1L1 R1L2 R2L 1 K2L2 K3 Z R1 R2 LI L2 k)i l Kit. 2 R2L 1 R2L2 R3 770695P 770 a9 7J i V/06S`VP 770701k 770703k 770705k 770707k 770709k 7 7071 IF. 770713k 770715k 770717k 7/0/1 Vis 770721k 770723k 770723k 770727k 770/2S'k 770731k 770733k 770733k 770737k 770 73S'K 770741k 770743K 770743k 770747k 770749k 770751k 770753k 770753k 7/0/37k 770739k 770761k 770763k 7707631 770767k 770769k 770771k 770773k 770773k 770777k 770779k 770781k 270783k 770785K 770787k 770789F. 770791k 770793k 770795k 770797k 770799K P 93 P 97 l.i 99 P101 P103 P) 03 Ml 07 PI 09 Pill PI 13 P x 15 P.U7 PI 19 PI 21 P) 23 p:c23 I.i i 2 7 P129 PI 31 D133 Pi 33 Pi 37 PI 39 D141 P) 43 11143 Pi 4 7 PI 49 P151 P3 53 Pi 33 PI 57 PI 59 P161 PI 63 PI 63 PI 67 PI 6 9 D J 71 PI 73 PI 75 PI 77 PI 79 D181 PI 83 D185 PI 87 111 89 P i 91 [1193 PI 95 P.1V7 PI 9V CO) (0 ) (0) (0 ) (0 ) <01 CO) CO) (0) CO) CO) CO) CO) (0) (CO (0) C0) CO) CO) CO) (0) CO) CO) (0) (0) CO) (0) (0) (0) CO) CO) CO) CO) CO) CO) CO) CO) CO) (0) CO) CO) CO) CO) CO) CO) CO) CO) CO) C0) CO) CO) CO) 139 139 139 1 4 ,i 14 T 141 141 141 143 143 J 43 143 143 143 143 145 145 1 45 147 147 147 147 147 149 149 149 149 ] 49 1 31 13) 151 151 1 51 1 53 153 153 153 3 53 153 ] 55 155 155 133 137 157 157 157 157 159 159 15V 139 159 R2L 1 R2L2 k`3 / R1 R2 LI L2 R i 1. .1 KILE R2L1 R2L2 K3 7 R'l R2 11 1.2 Rll. J R1L2 R2L1 R2L 2 R3 l R1 R2 13 L7 K.CI.1 R1L2 R2L1 K2I 7 k-7. 7 R1 K2 1i 1. 7 KJLi R1L2 R2L1 K21.7 K3 Z R1 R2 LI L2 Rl L3 k:il.7 K2L ) R2L2 R3 R&S 134259 AN3TX 20 (CONT. 6) LV 1 NCI SS REFERENCE LI STING - IOC - AY NCI. 1 25 Li 11- KLI kkFrkkNII Bill- " 1'1 MALES FEMALES ;gmr 708028 >08048 708068 7080BD 708ion 7081 28 708148 703160 7 0 818 D 70820D '708228 '70824 [i '708268 708288 '70830D '70832D '708348 '70B36D >7083BD >70B40B >708428 >708448 7708460 77084BD >708508 7708520 770B54H 7708560 7708580 7708608 770B620 77OB640 7708660 7708680 7708708 7708720 7708740 770B76D 7708780 7708800 7708820 7708840 7708860 7708880 7708900 7708920 770H940 EXF'R t *1 E4 E6 18 k 10 F 12 E 14 E 16 F 18 E 20 F ""l "1 E 24 E 26 F 28 K 30 E 32 E 34 36 E 38 E 40 E 42 E 44 E 46 E 48 E 50 E 52 E 54 E 56 E 38 E 60 E 62 E 64 E 66 E 68 F 70 E 72 F 74 E 76 k 78 F 80 F 82 F 84 F. 86 F 88 F 90 F 92 F` 94 SGI-' CAGE" (0) CO) CO) (0) (0) CO) CO) CO) (0> ' (0) CO) CO) CO) (0) CO) CO) CO) CO) (0) (0) CO) CO) (0) CO) CO) CO) CO) (0) c0) CO) CO) CO) CO) (0) (0) CO CO . CO) CO) CO) CO) CO) CO) (0) CO) (0) CO) 162 162 167 167 162 1 64 164 164 164 164 166 166 166 166 166 1 68 168 168 168 168 170 170 170 170 170 172 172 177 172 1 72 174 174 174 174 374 176 1 76 176 1 76 176 178 17H 178 178 178 180 180 FORK 7 R) Kl! LI L2 K1L.1 K-1I..2 R2L1 R2L2 R3 7 R1 R2 LI L2 R1L1 R1L2 R2L1 R2L2 R3 7 R1 R2 LI L2 R1L.1 R1L.2 R2L 1 R2L2 R3 7 R1 R? 11 12 Kl LI R1L2 R2L1 R2L2 K3 7 kl K2 L1 L2 R1L1 R1L 2 I.DMK 770801): 7708038 7708058 7708078 7708098 7708118 7708138 7708158 7708178 7708198 7708218 7708238 7708258 7708278 770B298 770B318 7708338 7708358 7708378 7708398 7708438 770B438 7708458 7708478 7708498 7708518 7708538 7708558 7708578 7708398 7708618 7708638 7708658 7708678 7708698 770B718 7708738 7708758 7708778 770B79I* 7708818 770HB38 7708858 7708878 7708898 7708918 7708938 hXPR E1 E3 E5 k7 k9 E 11 E 13 E 15 K 17 E. 19 E 21 E 23 E 25 E 27 fi 2 S' E 31 E 33 E 33 L 37 E 39 E 41 E 43 E 45 F. 47 E 49 F 51 F 53 F. 33 k. 57 F 59 F 61 F. 63 F 63 b 67 F 69 E 71 F 73 F 73 k 77 F 7V F. 81 F 83 E 85 F F(V k By t 91 F 93 SGF- C.AGF CO) CO) (0) CO) CO) CO) (0) CO) CO) CO) CO) CO) (0) CO) CO) (0) CO) CO) CO) (0) (0) (0) (0) CO) (0) (0) CO) (0) (0) (0) CO) CO) CO) CO) (0) (0) <0) CO) (0) (0) CO) (0) (0) CO) (0) CO) CO) 161 1 A) c 6.1 161 161 163 1 63 163 163 163 165 165 1 AS 165 165 167 167 167 167 167 169 169 169 169 169 171 171 171 171 1 71 173 173 1 73 173 173 173 175 175 175 1 75 17/ 1 77 1 77 17 7 177 1/9 1 79 k iKK` / R-l k? Ll L? R1L1 R1L2 R2L1 R2L2 R3 7 R1 R2 Ll L2 R1L 1 R11. 2 R2.L1 R21.2 R3 2 R1 R2 LI L2 R1L1 R1L2 R2L1 R2L2 R3 / R'1 K7 I1 L2 R11.1 R1L2 R2LI ft 21. K3 / Kl K2 L1 l2 Kill Kit 7 -_\-!,TX 20 (COST. 9) ' N | s s REF ft EMC E L I STING - )M - NLl. i:-:-. 0 ZERO REF FlftF N l: F I'MI " i HOLES ift XPR I9 6 0 -tR'BD /OOD ?02D *04H 706D ?osr> ?10D 911!!' 914D 91 60 9181 9200 92.20 9240 9261 9291 9301 932D 9341) 936H 9381 9401 >9420 9440 9460 >9480 >9500 >9521 >954 [i >9560 >9580 >9600 19620 J9640 29660 '968 0 39700 J9720 39740 09761 09780 09B0I 09820 09840 09860 09880 09901 09920 09940 09960 09980 10000 E 9c. F 90 F 1 00 t 102 104 t ] 06 F J OP F .1 I 0 E 312 E114 .316 tllP F 120 FJ 22 E124 F" 1 26 F ] 38 E 130 FI 32 134 136 fc 1 38 F ] 40 Fl 42 F.144 h 146 F 1 48 F 150 F 152 F 1 54 F 1 56 F 158 F 160 F162 F 164 F 1 66 F i ie El 70 El 72 F 1 74 F 1 76 F 1 78 E180 fc 182 fc ] 84 E186 F 188 fc 1 90 fc J 92 fc 194 F' 196 F 198 F 300 SKF CAGE (0> 0> <0> CO) CO) (0) CO) co> CO) CO) (0) (0> CO) (0) (0) CO) < 0) (0) (0) CO) CO) CO) CO) CO) CO) CO) CO) (0) CO) CO) CO > CO) CO) CO) CO) CO ) CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) C0) CO) co) co) 1 HO I HO 180 182 183 182 183 1 R2 1 H4 1H4 1B4 184 1 84 1 86 186 186 1 86 1 86 1 88 1 Hfcl 1 HH J Rb 1 88 .190 190 i yo 1 90 ) 90 1 93 i y:> 1 V? 1 92 193 194 1 V4 194 1 94 I 94 196 1 96 196 1 96 1 96 i9e ]>-! 1 v;< 1 VH 198 300 .'00 300 300 200 F ARK K2L I ft 2 L 2 R3 7 KM. R2 Ll L2 K1L .1 Kit.2 R2L1 R2L2 K3 7 K1 R2 U l2 KI L 1 K1L.2 R2L1 R2L 2 K3 7 K1 R2 l1 1,2 Kll 1 K1L2 R2L 1 R2L2 R3 7 Rl R2 11 t2 Kll 1 R1L2 R2L 1 ft21 2 K3 7. K3 ft 2 Ll 1.2 RU 1 ftll 3 Kill 1 R2L2 R3 ffhales CtlHK fc' Xfc'ft SUP CAGE F AftK 7708950 7708978 7708990 77090ID 770903D 770905D 770907D 770909D 77091111 7709]3D 770915D 7 7 0 917 D 770919H 770921D 7709::3D 770935D 770927D 77092911 77093ID 7709330 770935D 770937D 770939D 770941D 770943D 770945D 7709 4 7 D `709490 77095 J Li 770953D 770955D 770957D /7095VD 770961D 770963D 770965D /70967H 770V69D 770971D 770973D 7709750 7709/70 770979D 770981D 7709H3D 7/09850 770987D 7709B9D 77099)D 7709930 7709950 7709970 7709990 fc ys t. 97 fc 9V E 101 103 E1 05 E 107 fc 109 E.l 13 El 13 E 115 E. 11 7 fc. U V t.171 E 123 t 3 25 fc 137 fc 1 29 fc 3 31 El 33 fc. 135 t 1 37 fc 139 fc. 1 4 1 F143 F. 1 4 5 1- 147 fc 3 49 fc. 1 51 t.3 53 fc 1 55 fc 1 57 fc 3 59 161 E.) 63 fc 165 fc 3 67 E169 El 71 E 1 73 t 175 fc 3 77 El 79 fc 3 HI fc 183 El 85 1.187 fc. 1HV t 191 fc 3 93 El V5 fc 1 97 t 1 9V (0) C Ci i CO/ CO ) C0/ CO) (0) (0) CO) CO) CO) C0) CO i C0) COi CO) (0) C0) CO) ,C 0) CO ) CO) (0) COi CO) (0) (0) CO) CO) CO) CO CO) CO) (0) C () > CO) CO; 10) COi CO) (0) CO) CO) CO) (0) CO) CO) CO/ (0) CO) CO) CO) CO) 3 7v 1 179 18] I 83 3 83 3 8.1 181 183 183 3 82 1 83 182 185 185 3 85 3 85 185 3 87 1 87 3 87 187 iey 1 89 1 HV 18y 3 B9 1 FI 9 19) J Vl 193 3 91 3 91 3 93 193 193 3 93 3 93 1 95 3 V5 195 3 95 1 95 19/ 197 3 97 )y 7 1 97 ] 99 J 9V 1 99 1 VV 1 99 K7I 1 K.'l 2 ft 3 7 ft.1 ft".' L3 1 ~> ftlt 1 ft 11. 2 K71 1 F;?L2 ft 3 / ft'3 ft 2 L3 L : ftll 3 ft 1 L 2 ft3L 1 R2L.2 K3 7 ft) F:2 t3 12 K 11 3 ft) 1 2 K2L1 ft 21 2 K3 7 ftl F:7 13 1 KU3 K) L2 ft2Ll ft/L? ft 3 7 ft) ftl' Ll 1.2 Kll 1 KM 2 K2I 1 R2L2 ft 3 1AL MALES >- 500 lOTAI FhHAI fci, 500 i R&S 134260 R&S 134261 ,'0' 1 NO R05 S R E F E ft E N C E L1 -SAY NC. i: 6 0 hAL c r-t TOMR .'6000? rsoo04 780006 ?soooe 780010 *8001 2 TS001-S 780016 780018 780000, 780002 780004 780006 7S0006 7S0030. 7B0033 760034 780036 780038 780040 780042 78004 4 780046 780048 780050. 780052 780054 780056 7B0058 780060 780062 790064 780066 780068 780070. 780072 780074 780076 780070 780082. 780082 780084 780086 7B00B8 7B0090. 780092 780094 700096 780098 780100 Exf-R Ao A4 A6 A8 A 10 A A 14 A 16 A IB A 20 I* *1 l* 4 6 b8 6 10 6 12 tt 14 8 16 B 13 6 20 C2 c4 c6 c8 c 10 c 12 c 14 c 16 c IB c 20 p b4 P6 P8 10 n 12 [1 14 0 It I' 18 D 20 En E4 E6 E8 E 10 E 12 E 14 E 16 E 18 F 70 SGF (0) <0' (0) (O' (0' (O' (0) (0) (0) (0) i0) <0(0) (0) (0) CO) (0) (0) (0) (0) (0) (0) (0) (0) (0) (0) (0) <0> co> (0) (0) (0) <0> (0) (0) (0> (0) (0) <0> (0) (0) (0) (0) (0) (0) <0) (0) (0) (0) (0) CAGE "1 n 7 7 7 A A 4 A A 6 6 6 6 6 8 8 8 8 B 10 10 10 10 10 12 12 12 12 12 14 14 14 14 14 16 16 16' 16 16 18 IB 18 18 18 20 20 20 20 20 EARS 7 PI R2 1.1 t2 R 1L 1 R1L2 Rl'L 1 R2L2 R3 Z R1 R2 LI L2_ R1L1 R1L2 R2L1 . R2L2 HS. 2 R1 R2 LI L2_ R1L1 R1L2 R2L1 R2L2 F_ 2 R1 . R2 LI L2 R1L1 R1L2 R2L1 R2L2 R3. 2 R1 R2 LI L2 R1L1 R1L2 R2L1 R2L2 R3 TOT Al. MALTS 50 -TING FEMAl EB CO MR- EX PR SGR r A Lit E ARK 780001 780003 780005 780007 780009^ 7B001T 7B0013 780015 780017 7B0019_ 780021 730023 780025 780027 780029 780031 780033 780035 780037 780039 780041 780043 7B0045 780047 7B004.9 780051 780053 780055 780057 780059 78006*1" 780063 780065 780067 780062. 780071 780073 7S0075 780077 780079. 780081 780083 780085 780067 780089^ 780091 78'>093 780095 780097 780099 A1 A A 3t* A7 A9 A 11 A 13 A 15 A 17 A 19 81 B3 B5 B? 69 8 11 B 13 P 15 B 17 B 19 C1 C3 c c7 c9 c 11 c 13 c 15 c 17 c 19 D1 p3 p5 p7 n9 p 11 p 13 p 15 p 17 p 19 E1 E3 E5 E7 E9 E 11 E 13 E 15 E 17 E 19 (0) (0) (0) (0) (0) (0) (0) (0) (0) (0) (0) (0) (0) (0) (0) (0) (0) (0) (0) (0) (0) (0) <0> (0) CO) <0> (0) (0) (0) (0) (0) (0) co> (0) (0) CO) (0) CO) CO) CO) CO) CO) CO) (0) CO) CO) CO) CO) CO) CO) 1 I ft 1 1 F2 1 L1 1 i" 3 R1L 1 3 R1L 2 3 R2L1 1 R2L 2 3 R3_ r( 7 5 R1 5 R2 t LI L2_ 7 R1L1 7 R1L2 7 R2L1 7 R2L2 7 R3_ 9T 9 Rl 9 R2 9 LI 9 L2_ 11 RlLl 1 1 R1L2 11 R2L1 11 R2L2 11 R3 13 Z 13 Rl 13 R2 13 LI 13 l;_ 15 RlLl 15 R1L2 15 R2L1 15 R2L2 15 R3 17 1 17 Rl 17 R2 17 LI 17 L^_ 19 RlLl 19 R1L2 1 9 R2L 1 19 R2L2 19 R3 total females = 50