Document 8RDveN5bp2MOpYdxZ622Yd7ZZ

ro yy -- - ycM ' yixic/rr Cc p O -- (=> 3x. r>Vvj^Lh^t tiy. <6hjJ\ Clvo Institutes TNO Report No. V 83.285/291099 LIFESPAN ORAL CARCINOGENICITY STUDY OF VINYL CHLORIDE IN RATS (Final Report) If tf .9*7 R&S 040699 T/ ~^ny^ vet RECEIVED OCT 29 1984 YC TECH CTR civo institutes tno 1 * A: .. /^ Netherlands organization tor applied scientific research / TerKt-v'* GCD division for nutrition and food r*A(th tno p.o. box 360 3700 AJ zeist Netherlands Report No. V 83.285/291099 LIFESPAN ORAL CARCINOGENICITY STUDY OF VINYL CHLORIDE IN RATS (Final report) Authors : Dr E.P. Til H.R, Immel Dr^V.J. Feron ,_t At the request of : Verband Kunststofferieugende Industrie E.V., Frankfurt am Main, B.R.D. Project number : B 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 Date : Dr A.P. de Groot : September, 1983 Total partial publication ofinis report without written assent is not allowed ?3j toooo Otfll i R&S 040700 V 83.285 CONTENTS SUMMARY 1. INTRODUCTION 4 2. MATERIAL AND METHODS 2.1 Materials 2.2 Preparation and administration of the diets 2.3 Animals and housing 2. A Identification of the test system 2.5 Experimental conduct 2.6 Statistical analysis 2.7 Contributors 2.8 Deviations from the protocol 3. RESULTS 3.1 Analysis of VCM in the diets 3.2 Levels of VCM in the faeces 3.3 Actual oral exposure levels of VCM 3.A Symptomatology and survival 3.5 Body weights 3.6 Food intake 3.7 Haematology 3.8 Glutathione levels of the liver 3.9 Pathology A. DISCUSSION 5. CONCLUSIONS page 4 7 8 8 9 10 11 12 17 18 18 19 19 19 19 20 21 21 22 22 22 25 27 33 CO O O N| O I V 83.285 CONTENTS 6, REFERENCES 7. AUTHENTICATION 8. RETENTION OF RECORDS AND SPECIMENS STATEMENT OF CLP COMPLIANCE TABLES FIGURES ANNEXES page -3- 28 30 31 32 33 70 71 3D W o o Nl o ro V S3.285 -A - SUMMARY 1. The oral carcinogenicity of vinyl chloride monomer (VCM) was examined in a lifespan study (149 weeks) with five groups of Wistar rate, each consisting of 100 males and 100 females, except for the top-dose group a which comprised 50 males and 50 females, VCM 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 The diet was provided daily for a period of 4 consecu tive hours, whereas food was withdrawn during the other 20 hours.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 og VCM/kg body weight/day. An extra control group of 100 rats/sex was housed in a separate room. Additional satellite groups of 10 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 and 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 glands. ,, , 2. General health, behaviour, body weight and food intake 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 was 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 the top-dose group was slightly higher than in the lower dose groups and the controls. 4. Thrombocyte count, prothrombin time and liver glutathione levels did not show treatment-related differences among the groups. 040703 00 Ho CO V 83.285 I L ^ *, - 5- 5. A clearly higher Incidence of grossly visible, tumourous, liver nod ules was found In both males and females of the top-dose group than In any of the other groups. Moreover, In females of the top-dose group the Incidence of hepatic cysts was considerably higher than In con trols. 4 6. 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 compared to the control group. Moreover, a hepatic angiosarcoma was found In one male and two females of the top-dose group, whereas no such tumours were encountered In any of the other groups. The number of animals bearing foci of cellular alteration in the liver was also statistically significantly Increased In females of the mid-dose group as compared to controls. In addition. In females but not in males, the incidence of basophilic foci of cellular alteration in the liver was statistically significantly higher In both the lowand the mid-dose group than In the control group. R&S 040704 7. There was no evidence of VCM-feeding affecting the Incidence of ab dominal mesotheliomas or the type and Incidence of mammary gland tu-L mours. No Zymbal gland tumour was found. 8. It was concluded that under the conditions of the present experiment: - VCM at a level of 1.3 mg/kg body welght/day induces neoplastic and non-neoplastic changes in the liver of rats, - VCM at a level of 0.13 mg/kg body welght/day may lead to more female rate bearing foci of cellular alteration In the liver, - VCM at levels of 0,014 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 VCM/kg body welght/day is a "no-observed-adverse-effeetlevel" with respect to the Induction of tumours in rats. V 83.285 ^ L.' -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 that the cancer risk of a likely maximum oral daily Intake of 0.1 pg VCM per person per day can be practically neglected. R&S 040705 V 83.265 LIFE-SPAN ORAL CARCINOGENICITY STUDY OF VINYL CHLORIDE IN RATS 7 1. INTRODUCTION From a previous life-span oral toxicity study of vinyl chloride monomer (VQl) in rats with dose levels of 1.7, 5.0 and 14.1 mg/kg body velght/day (Feron et al., 1978; 1981), It appeared that hepatic angiosarcomas occurred at dose levels of 5.0 og/kg body weight/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 weight/day. Moreover, there was some evidence that VCM en hanced the formation of lntra-abdomlnal mesotheliomas and adenocarcinomas of the mammary gland. In females of the low-dose group there was still a high incidence of liver-cell tumours, via. 28/58. Thus, this study showed a) that VCM is a carcinogen in rats when administered orally, and b) that the "no observed-adverse-effect level* of VCM in rats with respect to the Induction of tumours was lower than 1.7 mg/kg body velght/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, vis. nominally 0.017, 0,17 and 1.7 mg VCM/kg body weight/day (actual oral exposure levels 0.014, 0.13 and 1.3 mg VCM/ kg body weight/day) and two control groups. Based on the pathological findings of the previous long-term study, hlstopathologlcal 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. R&s 040707 V 83.285 2. MATERIAL AND METHODS - 8- 2.1 Materials 4 Vinyl 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 S- 99.97 wt Z min,; acetylene 4 2 pl/1 (gas); mono-vinylacetylene ( 15 jil/l (gas); 1,3-butandiene f 10 jil/1 (gas); methyl chloride $ 75 jil/1 (gas); ethyl chloride 6 50 ;il/l (gas); chloro- prene i 1 ft1/1 (gas); 1,1-dlchloroethane i, 1 pl/1 (gas); 1,2-dichloroeth- ane < 20 jjl/1 (gas); acetaldehyde & 5 mg/kg; hydrochloric acid 4 1 mg/kg; Iron 6 0.5 mg/kg; water 6-100 mg/kg; evaporation residue 6 10 mg/kg. FVC-nowder. commercial name Carina S 65-02, was supplied by Shell Nederland Chemle, Pernis, The Netherlands, In closed steel barrels. The particle size distribution (by weight), specified by the supplier, was: 0.1 Z max. > 300 pm; 4 Z max. > 200 pm; 90 Z 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 than 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 a few minutes before mixing with each diet. R&S 040708 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, Jn that study the dietary levels of VCM were obtained by incorporating into the diets 10 2 PVC-powder with varying proportions of VCM-contalning and "VCM-free" powder. In the present study, the diets were prepared to a total concentration of PVC-powder of 1 Z only. The proportions of PVC-powder with or without VCM for each of Che diets are specified below: group PVC-powder In the diet (I) PVC-powder con PVC-powder taining about 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 VCH. The VCM-content was lower than 0.2 ppm Diets were prepared dally - 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 detection 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 o40709 V 83.285 - 10 - As in the previous study, the powdered diets were available to the rats each day for a period of four hours (generally between 10.00 a.m. and 02.00 p.m.), in 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-May 1981, September-November 1981 and January-March 1982 are given in Annexes 14 to 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 TNO, Zeist, The Netherlands. The animals arrived on August 14, 1979, when they were about 25 days old. The body weight range 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, viz. the 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 the satellite groups, which received the same diets as the rate 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 start of the study. None of the other reserved were used. R&S 040710 V 83.285 11 The rats were housed under conventional conditions, five rats per sex per cage. In suspended stainless steel cages (17 x 44 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 cr 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 rats of the control-, low-, mid- 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 was kept at 23+1 *C, the relative humidity was 40-80 1 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 days, 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) was fitted with a letter and colour code. A table showing the group letter, colour code, exposure level and number of animals is given on the next page. group colour letter code A white B blue C green D red E brown nominal actual exposure levels exposure levels (mg VCH/kg b.w./day) 00 0.017 0.014 0.17 0.13 1.7 1.3 00 number of rats main groups satellite groups study no. 125 study no . 126 males females males females 100 100 10 10 100 100 10 100 100 10 10 10 50 100 50 10 100 10 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 - one right plus one left R^ one right RjLj " one right plus two left *2 " two right one left - two right plus one left RjLj ".two right plus two left Lj " 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 reference num ber 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 May 20 or 22, 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 June 24 to 29, 1982, when about 80 Z of the control-, low-dose and mid-dose rats had died. The following determinations were made: V 83.285 a) Analyslsof VCMlp the diets 13 - R&S 040712 The VCM content of freshly prepared diets was generally determined once every fortnight. Burlng the course of the study, 72 determinations for each dose level were carried out. At the same time the VCM content of the 4 PVC-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 A 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 feedlng-perlod 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 2A 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 VCM 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. R&s 040713 V 63.285 14 - The VCM Intake (mg/kg body uelght/day) was 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). Both the rate of evaporation and the rate of food intake were assumed to be linear within each hourof the feeding-period. The actual VCM Intake for each group during each of 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 welght/day) was calculated based on an average of 36 g wet faeces/100 g foodconsumed. The amount of VCM in the wet. faeces (mg/kg body welght/day) was calculated by multiplying Che 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 welght/day, was substracted from the actual oral Intake of VQi (also expressed In mg VCM/kg body welght/day) to find the actual oral exposure level of VCM. d) Clinical signs (main groups) 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) Body 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 weight of the animals of each group was calculated. V 63.285 f) Food consum2>tion;_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 46, 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 4, 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) ft&S 040714 Samples of blood were collected from the tip of the tall 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/sex/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(Na2) solution. A part of the homogenates was centrifuged at 2800 rpra for 5 minutes. The non-protein bound sulfhydryl ('glutathione') con tent of the supernatants was determined with 5,5'-dithiobls-(2-nltrobenzolc acid) by means of an AutoAnalyaer. V 83.285 1) Pathology 16 - R&S 040715 All males 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 time a mortality of about 80 2 had been reached the control, low- and mid-dose groups. A thorough autopsy was also performed on rats found dead or killed In ex- tremls. 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 mammary 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 site 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, site 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 Z aqueous neutral phosphate-buffered formaldehyde solution: adrenals aorta axillary lymph nodes brain caecum cervix coagulating glands ovaries pancreas parotid salivary glands preputial glands pituitary prostate seminal vesicles V 83.285 - 17 - colon skeletal muscle duodenum epididymides skin spinal cord exorbital lachrymal glands spleen eyes sciatic nerve sternum (with bone marrow) 4 stomach heart Ileum submaxlllary salivary glands sublingual salivary glands jejunum kidneys testes thymus liver thyroid lungs trachea mammary glands urinary bladder mesenteric lymph nodes uterus nose Zymbal glands oesophagus all gross lesions Tissues required for microscopic examination were embedded In paraffin wax, sectioned at 5 pa and stained with haematoxylln and eosln. Histopgthology 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. R&S 040716 2.6 Statistical analysis Data on body weights were subjected to one-way analysis of (co-)varlance, followed by Dunnett's multiple comparison test. Thrombocyte counts and prothrombin time values were analysed by the Mann Whitney U-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. Grossly visible masses were evaluated by the Chi square test. V 83.285 2.7 Contributors Major contributions to this study were made by: Animal handling and: Hs. M.M. Andrlnga diet preparation J.M. Blom Ms. A. Dijkstra Ms. C.C.M. Fleer Ms. A.A. van Tuyl D.C. Veldhuysen Diet analysis : A, Schouten Clinical chemistry : Dr H.E. Falke J.F. Catsburg Biochemistry : Dr D.C. Leegwater Hlstotechnique : Ms. M.T. v. Rijswijk Ms. J. Wlsman Ms. A.P.M. v. Tuyl-Fonville Pathology Ms. N. Hagemeyer U.R. Immel Dr V.J. Feron Study director Dr H.P. Til . Study supervisor Dr V.J. Feron 18 - R&S 040717 2.8 Deviations from the protocol - At the request of the sponsor, the glutathione determinations in the liver were carried out after S and 18 months Instead of after 6, 12 and 24 months. - Prothrombin time and thrombocyte count in rats of the satellite groups after 24 months could not be carried out, because all rats of the sat ellite group had already been killed before that time. V 83,285 3. RESULTS 19 - 3.1 Analysis of VCM In the diets (tables 1 and 2) 4 The average VCM content of freshly prepared test diets appeared to be 0.49, 4.49 and 44.1 ppm for the low-, mid- and top-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 Z 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 Z for the low-, mid-, and top-dose groups, re spectively. 3.3 Actual oral exposure levels of VCM (table 4) The VCM contenc of the freshly prepared test 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 four-hour feeding-period (table 2), the rate of food intake (table 10) and the VCM content of the faeces (table 3) were known, 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). | R&S 040718 V 83.285 20 - When the actual oral Intake of VCM was expressed as percentage of the theoretical Intake, the values found were 81.5, 81.2 and 81.3 Z for males, and 80.1, 79.8 and 79.8 Z 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 Z. This percentage was in good agreement with the 80 Z 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 VCM. 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, stalred 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 was higher than In the other groups. This remarkable differ ence appeared to be caused by the 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 a& 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. R&S 040719 V 83.285 21 The cumulative mortality is given in table 6. Up to week 68 mortality was very low. Thereafter, mortality gradually Increased, more quickly in the 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 the 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). Males 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 040720 V 83.285 - 22 - 3.7 Haematology (table 11) Thrombocyte count did not show 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 4 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 4 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 size, 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. R&S 040721 i j j R&S 040722 V 83.285 - 23 - 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. 4 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 14) Increased Incidences of foci of cellular alteration, neoplastic nodules, hepatocellular carcinomas, liver-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 the long-term effects of oral administration of VCM in rats (Feron et al., 1981), no further discrlption of these alterations Is presented In this report. The morphology of VCM-lnduced hepatic angiosarcomas has also been described in a previous report (Spit et al., 1981). Table 14 also shows that In females, but not In males, of both the low-- and 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 than In the control group. V 83.285 24 - R&S 040723 There was no evidence that any of the other 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 cholangiofibrosis 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 hepatocytes 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, cholangiofibrosis and vacuolisatlon 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). Mammary glands (table 15) The number of females bearing a mammary 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) All intra-abdominal tumours found at autopsy were examined histologi cally. The numbers of mesotheliomas detected in rats of the different 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 Zymbal gland tumour was found. V 83.285 - 25 - 4. DISCUSSION In the present study no effects attributable to VCM were 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 VOI/kg body weight/day) used in the previous long-term oral rat study with VCM (Feron et al,, 1981). R&S 040724 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 in 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 Z; Feron et al., 1981). Similar incidences were also found ij females of control groups of three othdr 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 12 2, 3/20 or 15 Z and 17/99 or 17 Z. These data seem to Indicate that the Incidence of basophilic foci (9 Z) 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 Z, 1/20 or 5 Z, 5/69 or 7 Z and A/50 or 8 Z. Compared to these figures an incidence of 9/98 or 9 Z 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 0.01A or 0.13 mg/kg body welght/day may have resulted in more basophilic foci of cellular alteration in Che liver of female rats. The directive of the European Community (EC) of January 30, 1978 (78/142/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-directive 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 dally intake of VCM by humans mean in terms of cancer risk? V 83.285 - 27 Extrapolation of the results of the present rat study to humans using the linear extrapolation model and taking into account an 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 daily Intake is only 0.1 pg VCM/person/day, the estimated cancerrisk 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 might be (considerably) lover 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 (Gehring et al., 1979). Translated into practical terms this means that the cancer risk of a maximum oral dally intake of 0.1 fig 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 weight/day induces neoplastic and non-neoplastic changes in the liver of male and female rats, - VCM at 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. V 83.285 6. REFERENCES - 28 - Ministry of Agriculture, Fisheries and Food (1978). The Surveillance of Food Contamination in the United Kingdom: Survey of Vinyl Chloride Content of Polyvinyl Chlorldf for Food Contact and of Foods. The Second Report of the Steering Group 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, 428-430 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. Cosmet. 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 R&S 040727 Gering, P.J., P.G. 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 Owren, P.A. Thrombotest: a new method for controlling anticoagulant therapy Lancet, 1959, II, p. 754 Spit, B.J., V.J. Feron and C.F.M. Hendrlksen Ultrestructure of hepatic angiosarcoma In rats induced by vinyl chloride Exp. Mol. Pathol. 35 (1981) 277-284 V 83.285 - 29 - Squire, R.A. and M.H. Levitt Report of a workshop on classification of specific hepatocellular lesions in rats Cancer Res. 35 (1975) 3219-3223 INST. CIV0-T0X. AND NUTR. TNO 83-09-017AE 30 w O O 'si NJ 00 V 83.285 7. AUTHENTICATION This report was prepared by: 30 - (Study director) date H.R. Imnel (Pathologist) date: 2. $ - <&5 3 (Study supervisor. Pathologist) and approved by! Dr A.P. de Groot date: (Head Dept. Biological Toxicology) R&s 040729 V 83.285 6. RETENTION OF RECORDS, SAMPLES AND SPECIMENS - 31 All records concerning the dietary formula, daily observations, body weights, food intake, haematology, clinical chemistry and gross- and microscopical examinations an<! all other information relevant to the quality and integrity of the study have been filed in the archives of the . Department of Biological Toxicology together with the master copy of this report. Materials (samples of the test substance, wet specimens, blocks) 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 15 years, i.e. till July 1997. R&S O o Nl Co R&S 040731 V 83.285 32 - QUALITY ASSURANCE UNIT TNO - F.O. Box 360, 3700 AJ ZEIST, Netherlands STATEMENT OF CLP COMPLIANCE On : Lifespan oral carcinogenicity study of vinyl chloride ln'rats Report no.: V 83.285/291099 Dace : September, 1983 The study was carried out under conditions of good laboratory practice. Wichln 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 (1) 22 and 25 February 1980 (1) 12 May 1980 (1) 12 November 1980 (1) 17 November 1980 (2) 23 January 1981 (1) 3-4 March 1981 14-15 May 1981 (2) (3) 18 August 1981 (1) 27 October 1981 (3) 22 January 1982 (3) 12-13 May 1982 (2) 24 June 1982 (1) 8 July 1982 (3) 8-11 March 1983 < 2) 10 March 1983 (1) 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 1981 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 Draft report audit: 10-11 March 1983 24 March 1983 Final report audit: 20 September 1983 (1) ^ yv- y Drs S. van Straten Quality Assurance Manager 20 September 1983 date: ^ S'3 R&S 040732 V 83.285 - 33 - 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 diets11 Group A B C D E Intended oral exposure levels of VCM (mg/kg b.w. /day) 0 0.017 0.17 1.7 0 Dietary level of VCM (ppm) MEAN SEM N < 0.1 0.487 4.487 44.07 <0.1 0.013 0.097 0.926 72 72 72 72 65 ^ The VCM concent of the PVC-powder with VCM was on average 4582 + 98 ppm. CIVO/TNO STUDY NO 125 CARCINOGENICITY STUDY IN RATS WITH VCM IN THE DIET TABLE 2 LOSSES OF VCM FROM THE DIET DURING A PERIOD OF FOUR HOURS Group Intended oral exposuire levels of VCH (mg/kg b.w,./day) VCM: levels in the diet during the four--hour feeding-period at different points of time expressed as ppm^ expressed as percentage of tp C0 *3 e4 (1 *2 (3 l4 A 0 < 0.1 < 0.1 < 0.1 < 0.1 < 0.1 B 0.017 0.48 0.37 0.32 0.27 0.28 77 67 56 58 0.03733 0.039 0.024 0.021 0.016 C 0.17 4.2 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 1.94 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 (tg) and further from the feeders In the cages of the rats one hour (t^), two hours (t2)> three hours (t-j) and four hours (t^) after preparation ^ The values are the means of 7 different analyses. ^ Standard error of the me1"1. SSfcf ClVO/TNO STUDY NO 125 CARCINOGENICITY STUDY IN RATS WITH VCM IN THE DIET TABLE 3 LEVELS OF VCM IN THE FAECES Croup A B C D E Intended oral exposure levels of VCM (mg/kg b.w./day) 0 0.017 0.17 1.7 0 Level of VCM (ppm) found in the faeces 2k hours after the start of the feeding-period^ MEAN < 0.1 0,26 2.66 23.6 < 0.1 SEM 0.028 0.273 2.968 N 5 5 5 5 5 ^ Faeces were collected by squeezing the lower part of the rat's abdomen; they were immediately submerged In 10 ml ethylacetate and then stored at 6 *C In a closed vessel until analysis for the VCM content. C1V0/TN0 STUDY NO 125 CARCINOCENICITY STUDY IN RATS WITH VCH IN THE DIET TABLE 4 DESIGN AND ACTUAL DOSE LEVELS OF VCH IN RATS FED DIETS CONTAINING PVC POWDER Croup A B C D E Intended oral exposure levels of VCM ppm mg/kg b.w./ in diet /day 00 0.34 0.017 3.4 0.17 34 1.7 00 Actual dietary VCH levels at the start of the feeding period 1 (ppm) 0 0.49 4.5 44.1 0 Theoretical oral intake of VCH2) (mg/kg b.w./ day) 0 0.022 0.21 2.1 0 Actual oral in take of VCM35 (mg/kg b.w./ day) 0 0.018 0.17 1.7 0 Actual ora] ex posure level of VCH*0 (mg/kg b.w./day) 0 0.014 0.13 " 1.3 0 ^.Average dietary VCH contents determined Immediately after preparation of the diets. ^ Assuming no loss of VCH by evaporation from the diets (see also table 1). Taking Into account the loss of VCH from the diets. ^ Oral Intake of VCH diminished by the faecal VCM. 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. sezofro s$b CIVO/TNO STUDY NO 125 CARCINOGENICITY STUDY IN RATS WITH VCM IN THE DIET TABLE 5 CROSSLY VISIBLE OR PALPABLE SUPERFICIAL OR INTRA-ABDOMINAL HASSES OBSERVED IN THE COURSE OF THE STUDY Group Intended exposu re levels of VCM (mg/kg b.u./day) Total no. of rats at start No. of rata with masses Total Non-survivors HALES A0 B 0.017 C 0.17 D 1.7 E0 100 100 100 50 100 35 45 36 18 32 40 31 17 14** No., of masses Total disappeared In course of time 55 10 55 10 42 7 23 6 16 1 FEHALES A0 100 68 / 59 B 0.017 100 80 69 C 0.17 100 63 49' D 1.7 50 38 36 E0 100 48** 48 112 119 115 87 79 3 8 6 2 4 STATISTICS: CHI-square test: ** P < O.Ol, *** P < 0.001 u i &mo syy mi-le h MALES 0 If [i A DA BAY DAY DAY DAY DAY DA I DAY [I A DAY DAY d i r DAY DAY DAY DAY DAY DA t DAY DAY [i A Y DAY DAY DA Y DAY [i A Y r. no 56 81 112 1 -VO US 1 ? T1 A 252 280 7 Aq 3 2-6 3 ' 392 -120 -t 'IS *? 6 501 5 32 560 5SS 616 61 1 i 7T 700 72 6 MORTALITV COHTROL INCIDENCES A 03 A nr A o:-: ll l 13 l IX i IX l 13 l IX l IX l IX i IX . i it; A AV A 23 A ay A 3.Y A 43 4 43 A i:; \ 13 t 43 43 4 43 i iy t 1 113 13 1 33 13 133 0.017 MG/KG 1 MCI DEUCES 0 03 A 03 0 03 l 13 l 13 l 13 l 13 l 13 i 13 l 13 l 13 l 13 i 13 l 13 l 13 i 13 l 13 l 13 4 43 43 43 A 73 0 83 8 83 1 2 123 15 153 15 153 1 5 153 .. 0.1 7 MG/KG IHCIDEHCES 0 ny 0 03 A 03 0 03 0 03 0 03 0 03 . 0 03 0 03 0 03 0 03 1 13 I 13 1 13 2 23 2 ny 3 33 3 33 4 43 5 53 4 43 y 73 7% 7 73 e 83 O 93 1 3 133 14 143 1 . 7 MG/KG IHCI DEUCES A 03 n 03 r, 03 A 03 0 03 0 03 0 03 0 03 0 03 A 03 A 03 0 03 0 03 0 03 0 03 0 03 0 03 0 03 1 23 1 2*i n 43 A 43 A UZ A fey 3 63 4 83 A 143 10 203 statistics: SHER EXACT PROBABILITY TEST 1 PC0.05 " P<0.01 ' PCO,001 TWO SIDED COMTRL'L ihcideuce: a r*\ V 03 7 03 A A A Ay A 07 A v. A f\ 03 0 03 A O'. 0 T. A Ay A ny A A r. y A nv V oy O oy C cy uy * 7 *. i 1 0*. 1 3 12 3 12LQPO ssa TABLE 6 MORTALITY (CONTINUED 1) hALE S L' H Y 7S4 [1H Y O 1 o DAY 3 -t 0 DAY Sie [i A Y OO f. [i A i ?'l [i /1Y O ^ D A Y 3 grt DAY 1 008 DAY 10 34 CONTROL INCIDENCES 15 152 31 212 28 23% 31 312 38 3B2 44 4 4 2 53 ` 5 32 58 592 72 ir/ 80 B02 0.017 MG/KG INCIDENCES 17 172 25 252 28 2 82 34 342 4 3 4 32 51 512 54 542 43 432 49 492 80 B02 0.17 MG/KG INCIDENCES 14 23 2163 2v 28 282 34 342 38 382 44 442 53 532 59 592 72 722 82 822 - 1 .7 MG/KG INCIDENCES 1 5 30 2 18 36 2 n'i i'l v 24 48 2 2 4" 482 2 9' 562 32** 64 2 35* * 70 2 39 * ` * 78 2 42* " 84 2 * statistics: FISHER EXACT F'ROPAB IL I TY TEST P<0.05 * P<0.01 * *' PC0.001 TWO SIDED CONTROL INCIDENCE: 3? * " ' 3 **_ A - * ' i **. 5 1 * *' \ 71 * * * '6* * * OO * 1 * 71 \ Ti1 03' X r,r, * < - 1 00 1 * * 100** 100 * * * 10 or. 100 *. 100`. 1 00 2 TABLE 6 MORTALITY (CONTINUED 2) FEMALES BAY BAY BAY PAY PAY BAY PAY PAY PAY PAY PAY PAY PAY PAY PAY PAY DAY PAY PAY PAY PAY BAY PAY PAY DAY PAY PAY PAY v 30 58 Pi 114 112 170 198 22i 251 282 310 338 3ii 391 4tt ISO 178 50i 531 5i2 590 i18 ili A71 702 730 758 CONTROL INCIDENCES 0 OX 0 OX 0 OX 0 OX 0 OX 0 OX 0 OX 0 OX 0 OX 0 OX 0 OX 1 IX 1 IX I IX 1 IX 1 IX 1 IX 1 IX v 2X t TV 4 IX 5 5X 4 &X 7 7X 10 12X 12 12X 15 15X 20 20X 0.017 MG/KG INCIDENCES 0 OX 0 OX 0 OX 0 OX 0 OX 0 OX 0 OX 0 OX 0 OX 0 OX 0 OX 0 OX 0 OX 0 ox 0 ox 0 ox 0 ox 1 IX T 2X T 2X 1 4X 1 IX 4 IX 5 5X 8 BI 8 8X 11 11 X 13 13X 0.17 MG/KG INCIDENCES 0 OX 0 OX 0 OX 0 OX 0 OX 0 OX 0 OX 0 OX 0 OX 0 OX 0 OX 0 OX 0 OX 0 OX 1 IX 1 IX 2 22 2 22 3 32 3 32 3 32 4 42 4 42 7 72 10 102 12 122 14 142 19 192 STATISTICS; rISHER EXACT PROBABILITY TEST * P<0.05 P<0.01 1.7 MG/KG INCIDENCES 0 02 0 02 0 OX 0 OX 0 OX 0 OX 0 02 0 02 0 OX 0 02 0 02 0 02 0 02 0 02 0 02 0 OX 1 22 1 22 4X n 42 2 4X 5 102 4 1 22 4 122 4 122 4 122 12 24 2 11 282 P<0.001 TUO SI BED CONTROL INCIDENCES 0 02 o OX r, OX 0 02 0 OX 0 OX 0 OX 0 ox 0 ox 0 ox 0 AV 0 ox 0 ox 0 ox 0 ox 0 02 0 ox n 02 0 OX0 OX 0 ox 1 IX t TV T TV T. Tv v 3X 7 TV 11 1 42 SSZOfrO SSd TABLE 6 MORTALITY (CONTINUED 3) F E MALE 5 PAY PAY PAY PAY PAY DAY PAY PAY DAY PAY 786 81 4 842 870 898 92 6 954 982 1010 1038 CONTROL INCIDENCES 20 202 26 262 28 282 30 30 2 35 35 2 41 417. 47 472 58 582 66 66 2 76 762 0.017 MG/KG INCIDENCES 1 5 152 25 252 32 32Z 36 362 41 4 12 51 512 60 602 66 662 72 722 77 772 0.17 MG/KG INCIDENCES 19 192 22 222 27 272 30 302 35 352 45 452 52 522 61 612 67 672 74 742 STATISTICS I FISHER EXACT PROBABILITY TEST P<0.05 * P <0.01 1.7 HG/KG INCIDENCES 15 302 16 322 19 38 2 21 422 24 482 28 562 32* 64 2 36* 722 43' ' 862 45' * 902 P<0.001 TWO SIDED CONTROL INCIDENCES 16 162 25 252 37 37 2 50* ' 50 2 58' ' 5BX 9 2 1 * * 92 v 100" 1002 100` 1002 100' " 1002 too*'1 1002 OfrZOfrO S'Sb 1 7TADLE MEAN BODY HEIGHTS (G) HALES CONTROL MEAN SEH N 0.017 HG/KG MEAN SEH N 0 .17 HG/KG MEAN SEH N 1.7 MG/KG MEAN SEM N CONTROL MEAN SEM N BAY BAY DAY DAY DAY DAY DAY DAY DAY DAY DAY DAY DAY DAT DAY DAY DAY DAY DAY DAY DAY DAY 'AY .. A Y DAY DAY DAY 'AY lAY 0 14 28 56 64 112 140 16B 196 224 252 280 308 336 364 392 420 448 476 504 532 560 58B 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 3B8.5 403.4 411.2 416.5 413.3 410.8 404.2 404,2 405.9 400.4 415.6 419.4 414.5 419.4 428 , B 431.1 427.8 425.4 419.8 0.5 0.B 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 98 97 96 96 96 96 94 94 94 94 89 88 87 53.2 96.9* 147. B 223*0* * 271.8 * 309.2 ' ' 340.2 357.0 370.8 377.7 381 . 2 400.0 403.3 406.5 409.1 403.6 397.3 399.0 403.3 395.2 407.5 414.9 412.5 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 1 00 100 100 99 99 99 99 99 99 99 99 99 99 99 . 99 99 99 99 96 ' 96 9 6 ' 93 ' 92 . 92 , 89 B6 85 , 85 , 53.2 95.8 148.9 232.0 263.3 323.B 343.0 364.7 372.8 3B5.7 387.8 405.2 407.7 415.0 411.7 407.4 402.8 404.7 410.9 404.0 416.1 422.6 418.4 425.2 431.5 437.8 431.4 428.0 423.3 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 4.6 100 100 100 100 100 100 100 100 100 100 100 100 99 99 99 98 98 97 97 96 95 94 93 93 93 92 91 87 86 54.3 98.2 149.2 224.5* 279.0 316.6 34B.8 364.1 384.3 389.3 387.3 40B.3 414.6 417.9 41B.6 419.4 411.9 413.1 414.8 409.2 420.6 426.7 423.5 429.3 43B.5 446.5 440.4 428.8 423.9 0.6 1.1 1 .8 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 4.8 4.8 5.1 5.0 5.0 5.5 5.3 5.5 5.8 6.1 5.8 7.4 6.9 50 50 50 50 50 50 50 50 *50 50 50 50 50 50 50 50 50 50 50 49 49 49 48 4B 47 47 46 44 40 51.5 89.3* * 141.7* 214,6* * 251.1* * 278.1 * * 297.5* * 314.5* * 318.5* * 339.8* * 351.4* * 365.0* * 37!.0* * 371,3* * 377.8* * 385.2* * 370.9* * 368.0* * 377.3* * 364.0* * 380.7* * 384.3* * 379.9* * 376.7* * 379.7* * 381.2 * * 365.9 * * 357.2* * 344.8* * 0.5 0.7 1.1 1.5 2.0 2.2 2.3 2.6 2.8 2.6 2.7 2.7 2.8 3.3 3.0 3.1 2.9 2.9 3.0 3.7 3.3 3.3 3.3 3.7 4.1 4.3 4.5 4 .9 5.5 100 100 100 100 100 100 100 100 100 100 100 100 100 1 00 100 100 100 100 99 92 92 92 92 92 90 87 83 78 71 I ;tatist ics : COUAR + OUNNETT TESTS ' p<0 . 05 P<0 .01 T WO SIDED < exp'. UNIT = AMHAL) alUL'i NU I LrtKLlNUbtnii.1 I I aiui't in uni j i in vlii in . n. uli TABLE 7 MEAN BODY WEIGHTS (G) {CONTINUED 1) MALES CONTROL MEAN SEM N 0.017 MG/KG MEAN SEM N 0.17 MG/KG MEAN SEM N PAY 786 DAY 812 HAY B 6 0 DAY 868 DAY 896 DAY 924 DAY 952 DAY 980 DAYIOOB DAYI036 DAY 1063 408.6 394.8 393.3 387.4 376.6 365.6 361.7 352,9 346.5 341.3 343.8 4.0 3.9 3.9 3.8 4.3 4.3 4.1 4.4 5.2 5.2 5.3 85 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 4.8 4.9 4.7 5.0 5.6 5.8 6.4 6.1 6.5 5.5 5.6 83 75 72 67 59 49 47 38 31 20 20 409.4 402,9 400.6 394.0 386.3 371,2 363.4 360.2 358.6 356.3 358.1 4.6 4.5 4.7 4.7 4.5 4.4 4.7 4.9 6.3 6.2 6.3 84 77 72 66 62 56 47 43 28 19 IB 1.7 MG/KG MEAN SEM N 414.9 403.5 398.3 393.7 382.5 380.0 368.6 363.3 354.2 366.7 368.7 6.7 7.1 5.9 5.7 6.1 7.8 7.7 8.0 7.4 7.8 7.9 35 32 29 26 26 21 18 16 11 B 8 CONTROL HEAN SEM N 322.9* * 302.5* * 297.9* * 299.3* * 288.1 * * 329.8 ----- -- 5.4 5.5 7.0 7. B 9.0 20.4 -- -- ---- 63 58 61 39 26 41. 0 0 0 0 0 STATISTICS! COOAR + BUNNETT TESTS P < 0.0 5 ' P<0.01 TWO SIDED . (EXP.UNIT = ANIMAL) 2t?i.01?0 S'SH TABLE 7 HE AN BODY WEIGHTS <G> (CONTINUED 2) FEMALES CONTROL MEAN SEH N 0.017 MG/KG MEAN SEH N Cl AY JAY JAY JAY JAY JAY .1A Y *ihY J AY JAY JAY JAY JAY JAY JAY )iVr' i AY i AY JAY iAY JAY 1A Y 'AY 'AY lAY 'AY JAY 'AY 'AY 0 14 30 5B 86 114 142 170 198 226 254 2B2 310 338 346 394 4 22 4 50 476 506 534 562 590 618 646 674 702 730 758 52,8 92.7 118.9 151.4 164.7 17B.0 183.3 188.4 189.6 193.9 199.0 202.9 210.2 209.2 211.5 212.1 207.9 210.1 214.0 213.8 21B>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 I.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. I 2.0 2.3 2.5 2.8 2.5 2,7 2.7 2. 7 2.7 3.0 100 100 100 100 100 100 100 100 100 100 100 100 99 99 99 99 99 99 99 98 98 96 95 94 93 90 89 86 80 52.9 92,5 120.2 153.4 166.2 175,2 1 S3.4 187.2 192.9 192.7 201,0 203.7 209.7 208.1 212.4 210.8 207 . 210.3 212.4 213.1 217.6 219.0 ^22,7 227.0 231.9 236.0 234.4 232.3 234.6 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 2 2.5 2.5 2.7 2.B 2.9 3.0 3.0 3.0 3.2 100 100 100 100 100 100 100 100 100 100 100 100 100 100 100 100 100 100 99 98 98 96 96 96 95 93 92 90 87 0.17 HG/KG MEAN SEH N 51.5 90,1 120.1 153.6 16B.6' 17B.5 1B4.0 190.6 190.B 195.6 201 .B 205.9 210.8 210.B 211.6 212.9 209.0 213.4 215.4 216.0 218.4 221 ,B 226.5 230.3 235.8 241.0 240.0 238.B 240.2 0.5 0.7 0.9 1.0 1.2 1 .3 I .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 2.2 2,2 2.4 2.5 2.6 2.7 2.B 2.7 2.9 100 100 100 100 100 100 100 100 100 100 100 100 100 100 100 99 99 98 98 97 97 97 96 96 93 91 B9 86 Bl TATISTICS : CO'JAR + DUNNETT TESTS * P<0 . 05 ** F'<0, 01 TWO SIDED 1.7 HG/KG MEAN SEH N 51.2 BB .5' 117.9 152.0 167.B 179.8 187.8 194.3* 194 .B 195.4 203.3 205.5 212.3 213.1 214.6 217.3 212.7 217.0 217.9 217.0 222.2 223.3 228.0 232.9 240.8 245.1 242.8 237.0 237.4 0.7 1 .2 1.3 1.5 1.8 2.0 2.1 242 2.4 ntt 2.4 2.6 2.8 2.B 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 4.2 50 50 50 50 50 50 50 50 50 ,, 50 50 50 50 50 50 50 50 49 49 48 48 48 46 44 41 44 44 38 36 CONTROL MEAN SEH N 52.8 90.4 ' * 117.4 160.0*' 167.3 176.3 182. B IBB.9 193.0 195.8 205.2 206.7 212.3 210.0 211.5 213.2 209.7 211.2 215.9 210.B 219.1 219.3 221.9 223.2 227.7 226.8* 224.0'* 21'- .8* * 219i0** 0.4 0,8 0.9 1.2 1 .4 1.5 1.5 1.6 1.6 1.6 1 .8 1.8 1.9 1.8 1.9 1.9 i .e 1 .9 2.0 2.0 . r* 2,1 2.1 '? . n n ^ '> 2.4 2.6 2.4 2.5 IOC 100 100 100 100 100 100 100 100 100 100 100 100 100 100 100 100 100 100 100 too 100 99 98 98 98 98 94 86 (EXR, UNIT - ANIMAL) r- bZOP0 S9y TABLE 7 MEAN BODY WEIGHTS <G> {CONTIHI)ED 3) F E H ft L E S CONTROL MEAN SEM N 0.017 MG/KG MEAN SEM N DAY 786 PAY 814 DAY 842 DAY 070 DAY 898 DAY 926 DAY 954 DAY 982 DAY 1010 DAY 10 3 8 DAY1042 230.7 224.0 224.2 222*7 217.9 216.6 212.3 211,1 212.7 212.4 214.9 2.8 2.6 2.7 2.9 2.9 2.9 3.0 3.3 3.4 3.6 3.3 GO 74 72 70 66 59 54 43 36 23 21 230.7 227.0 224.0 222 * 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 5.1 5.7 5.7 85 78 69 65 59 49 41 34 28 23 23 0.17 MG/KG MEAN SEM N 235.4 232.1 231.8 227.3 223.3 220.8 215.1 212.1 221.1 218.4 217.7 2.8 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 48 42 33 25 25 1.7 MG/KG MEAN SEH N 232.9 227.9 227,4 227. 1 227.0 224.6 217.2 194.2 208.6 214.2 213.2' 3.7 35 3.7 34 4.4 31 4.2 29 4.9 26 5,4 22 5.5 19 4 .B 14 3.7 * 7 3.1 4 ' 3.8 4 CONTROL MEAN SEM N 209 . B' 207.7* 203.1** 203.3" 200.6* * 217.6 --- -- -- -- 2.8 2.B 3.4 3,7 4.0 4.3 --- ---- 84 77 65 50 42 R 0 0 0 0 0 STATISTICS: CLUMP f DUNNETT TESTS P'0.05 " P<0.01 TWO SIDED (EXP.UNIT = ANIMAL) WZOfrO SSb IAPLE 8 MEAN BODY HEIGHTS (G>, HALES hay PAY PAY PAY PAY DAY PAY PAY DA : PAY [i A Y HAY PAY PAY PAY PAY PAY PAY PAY PAY PAY 0 IA 28 56 94 112 140 169 196 252 280 308 336 364 392 4 20 448 476 504 532 CONTROL MEAN SEM N 51.4 8? . 4 141.6 229*2 276.4 312.1 336.1 360.4 373.5 383.4 3G2.6 417.1 424.6 430.3 413.0 419.7 416.5 413.7 422.1 413.5 426.6 2.0 2.5 3.1 4.6 4 .B 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 io. e 9.6 11.6 10 10 10 10 10 10 10 10 10 10 10 5 5 5 f J 5 5 C 5 5 5 0.017 HG/KG MEAN SEM N . 53.4 94,0 146.3 232.8 288.7 325.9 354.7 374.5 384.2 385.5 395.4 425,5 434.3 439.9 440.0 436.0 432.6 434.0 421.8 405.4 423.2 2.1 2.4 3.2 5.4 6.3 7.3 10.2 8.3 9.7 12.2 9.7 8.5 12.2 13.2 13.0 11.9 10.4 10.9 6.8 10.2 9.2 10 , 10 10 , 10 10 10 10 10 10 10 10 5 5 5 5 5 5 5 5 5 5 0. 17 MG/KG MEAN SEM N 55.5 96.9 139,5 230.4 289.4 325.7 350.7 372.3 377.5 38B. 4 394.2 393.1 392.4 400.1 395.6 391.0 379.2 382.4 381.5 374.9 395.1 1 .5 2.0 9.1 4.3 6.5 8.8 9.6 10.6 9.5 6.5 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 9 9 4 4 4 4 4 4 A 4 4 4 ?TATI ST ICS ! A- 3PA + PUNNET! TESTS ECO.05 . P<0.01 TOO SIDED 1 .7 HG/KG MEAN SEM H 56.4 97.0 149.5 n?-* . o 280.3 321.0 341.9 360.8 379.6 391,1 394.1 412.3 415.1 4 18.4 421.4 421.8 413.2 417.9 421.8 408.7 422.7 1.0 1 .0 2.0 3.2 5.0 i. 6.3 6.1 4.9 5.5 7.0 10.1 10.0 8.2 8.9 8.4 8.8 8.1 7.5 4.8 6.8 10 10 10 10 10 10 10 10 *10 10 10 5 5 s 5 5 5 5 5 5 5 CONTROL MEAN 54.9 96 A 147.7 218.1 246.0* 270.3'* 2B6.9 * * 306.5' 314.6'* 337.6*' 340.6' 352.3'* 361.3" 364.1 ' ' 373 . 1 372.9' 359.2'* 356.0*' 359.6 ' ' 353.1 ` * 369.2' 1*5 -> *T A .7 S.A 9>C on 9,6 9*7 6.8 9.5 8.1 16.3 16.7 16.3 14.9 15.6 18.1 16.9 16.5 15.7 17.0 ( EXP .UNIT - AMI HAL % t st?i.ot?o s'sa STUDY HO 126 18-MONTH STUDY IN RATS WITH VCM IN THt UtET IABLE 8 MEAN BODY WEIGHTS (G > (CONTINUED) FEMALES CONTROL MEAN SEM N 0.017 MG/KG MEAN SEM N 0. 17 MG/KG MEAN SEM N DAY DAY DftY DAY DAY DAY DAY DAY DAY DAY DAY DAY DAY DAY DAY DAY DAY DAY DAY DAY DAY 0 16 30 SB 84 114 M2 MO 1 ?8 226 254 2B2 310 338 366 394 450 478 506 534 53.3 95.4 119.8 150.3 161.7 172.3 177.8 lf.B 185.9 187.7 195.3 195.7 197.3 198.7 200.7 203.1 198.8 201.4 208.0 20^ , 8 214.5 1.A 1 ,B 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 c. 5 5 5 5 5 5 5 5 5 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 n *>5 t 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,B 7.8 7.9 9.1 10 10 10 . 9 9 9 9 9 9 9 9 5 5 5 5 5 5 5 5 5 5 54.7 95.2 123.1 154.4 166.3 178.5 .185.9 190.7 191.0 196.7 201.3 195.1 202.2 199.5 198.2 198.7 199.6 201.0 200.7 202.2 204,3 1.6 1.9 2.4 3.1 3.3 4.1 4.6 4.9 4.2 5.0 4.3 7.2 8.3 6,9 8.1 8.4 8.1 7.5 8.1 8.8 9.0 10 10 10 10 10 10 10 10 10 10 10 5 5 5 5 5 5 -5 5 5 5 STATISTICS ! ANOVA + DUNNETT TESTS PC0.05 PCO.Oi TWO SIDED 1.7 HG/KG MEAN SEH N 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 5.5 6.1 6.0 5.6 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 5 5 CONTROL MEAN SEM 50.4 85.7* 111.8 152.2 160.9 173.6 179.6 184.8 186.2 190.4 198.3 204.7 211.4 211.6 215.4 214.0 210.7 212*2 217.2 214.2 223.0 1 .6 2.1 2.4 2.9 4.4 4.9 4.3 5.4 4.8 5.4 5.7 5.0 5.B 6.0 5.6 6.4 5.3 5.7 6.4 5.1 6.0 (EXP. 1UNIT = ANIMAL) 9frZ0t>0 SSd CIVO/TNO STUDY HO 125 CARCINOGENICITY STUDY IN RATS WITH VCH IH THE DIET TABLE 9 HEAN FOOD INTAKE Group A B C D E Intended intake levels of VCH (tng/kg b.w./day HALES 0 0.017 0.17 1.7 0 Food Intake (g/rat/day) In week 1 2 3 4 11 12 23 24 35 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 8.2 10.3 12.4 13.5 12.5 13.3 12.5 11.9 13.3 12.9 47 48 15.7 17.0 14.6 15.3 15.8 17.1 15.5 16.4 12.3 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 10.6 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 > l^lQp 0 CIVO/TNO STUDY NO 125 CARCINOGENICITY STUDY IN RATS WITH VCM IN THE DIET TABLE 9 MEAN FOOD INTAKE (CONT.) Group A B C D E Intended Intake levels of VCM (rng/kR b.w./day KALES 0 0.017 0.17 1.7 0 Food intake (g/rat/day) In week 60 73 74 85 86 97 98 109 110 16.2 16.5 16.6 14.4 16.2 15.2 15.9 16.1 13.6 .14.6 16.1 17.5 17.3 15.0 16.3 15.8 18.2 17.5 15.7 '14.9 12.6 13.3 13.3 13.012.4 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 FEMALES 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 V 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 8frZ0t>0 SSb C1V0/TN0 STUDY NO 125 CARCINOGENICITY STUDY IN RATS WITH VCH IN THE DIETS TABLE 10 FOOD INTAKE DUR] .0 THE FOUR-HOUR FEEDING PERIOD Croup Intended oral Intake Food Intake during the feeding--period over periods of one hour D levels of VCM (mg/kg b.w./day) expressed as g/rat expressed as X-tage 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 3-4 HALES A0 11.6^ 2.7 2.0 1.9 18.2 64 15 11 10 0.87 0.29 0.27 0.31 0.62 B 0.017 9.9 3.2 2.3 1.8 17.2 58 19 . 13 10 0.66 0.29 0.11 . 0.17 0.71 C 0.17 10.5 3.0 2.5 1.7 17.7 59 17 14 10 0.58 0.27 0.23 0.16 0.60 0 1.7 10.7 3.6 2.3 2.0 18.5 58 19 12 11 0.90 0.34 0.25 0.29 0.95 E0 8.8 2.6 1.7 1.6 14.6 60 16 11 11 0.66 0.24 0.17 0.15 0.69 overall A n 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 ove rail ^ Food intake figures are the means of 11 different determinations. ^ Standard error of the mean 60 18 12 10 55 17 14 14 54 20 13 13 53 19 15 13 55 18 14 13 54 19 13 14 54 19 14 13 l/* 6W0W) SSH T ABLE M MEAN MAEMATOLOGICAL FINDINGS RECORDED AT DAY 94,180 AND 347 ALES CONTROL MEAN SEM N 017 MG/KG MEAN SEM N 0.17 MG/KG MEAN sen N J.7 MG/KG MEAN SEM N CONTROL MEAN SEN N DAY 94 TMROMROC (10E9/L> 98?. ?7. 10 1036. 33. 10 1000. 23. B 996. 23. 10 866. 50. ? DAY 185 THROMDOC {10E9/L) 098. 17. 10 915. 43. 8 860. 45. 9 825. 51 . 10 879. 31. 10 DAY 367 THROMDOC <10E9/L) 946. ' 23. 5 855. 43. 5 895.1 21.' 4 930. 63. 5 672. 32. 5 DAY 94 F'TT (SEC) 55.0 1.7 10 52.9 1.5 10 53.0 2.2 9 50. B 1.3 10 54.2 2.3 8 DAY 183 F'TT (SEC) 36.8 O.B 10 35.2 0.4 6 37.2 0.6 9 37.0 0.7 8 39.0 0.5 10 DAY 367 F'TT (SEC) 49.3 1.4 5 50.1 2.7 5 50.5 0,6 4 50.2 1.8 5 52.5 1.2 5 statistics; mann'uhitney u--test * r-'<o.o5 1HROMBOC = THROMBOCYTE TIT = PROTHROMBIN TIME p<o.o2 ** r<o.oo2 two sided (exp.unit = animal) OSZOfrO ssy if ( UL I MO * table ii A *-0 MEAN HAEMATOLOGICAL FINDINGS RECORDED AT DAT 97,104 AND 366 FEMALES CONTROL MEAN SEM N ',>.017 MG/KG MEAN SEM N 0.17 MG/K'G MEAN SEM N 1 . 7 MG/KG MEAN SEM N CONTROL MEAN SEM N DAY 97 THROMBOC (10E9/L) 927. 19. 10 915. 27. 9 904 . 14. 10 863. 34. 10 94 3. 2 El. 10 HAY 184 THROMSDC (10E9/L) 972. 27. 10 1032. 34. 9 071. * ' 23. 9 1010. 22. 9 1010. 29. 10 DAY 366 THROMBOC (I0E9/L > 879. 26. 5 930. 106, 3 904. 24, 4 906. * 20. 4 929. 131. 3 DAY 97 F'TT < SEC > 41.6 0.0 10 40.2 0.0 9 43. I 1.6 10 47.4* * 2.1 8 51.3* * * 1.9 10 DAY 184 PTT < SEC) 33.2 0.5 9 32.0 0.4 9 33.0 1.0 8 36.2 1 .7 9 37.0* * * 0.7 9 DAY 366 PTT (SEC* 40.1 1.2 3 43.6 2.8 4 39.4 1.2 5 38.6 1.7 5 39.9 1.0 4 STATISTICS! MANN/UHITNEY U-TEST * P<0.05 THROMBOC = THROMBOCYTE m - PROTHROMBIN TIME r<0.02 * ** P<0.002 TWO SIDED (EXP.UNIT = ANIMAL > S?y CIVO/TNO STUDY HO 126 18-MONTH STUDY IN RATS WITH VCM IH THE DIETS TABLE 12 MEAN NON-PROTEIN BOUND SULFHYDRYL ("CLUTATHIONE") CONTENT OF LIVERS OF RATS FED VCM FOR 9 AND 18 MONTHS Croup A B C D E Intended exposure levels of VCH (mg/kg/day) 0 0.017 0.17 1.7 0 Non-protein bound liver SH In umol glutathlone/g liver In months 9 18 9 18 MALES 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 Under each mean the standard error of the mean ts given. In brackets number of rats If not 5. ZSlOtrQ s$y .'IVO/TNO iTUDY NO 125 CARCINOGENICITY STUDY IN RATS WITH VCH IN THE DIET Al'.l.F. 13. MACROSCOPIC PATHOLOGY Site and type of observations Initial number of animals Effective number of animals MODE OF DEATH: a. found dead b. killed in extremis c. terminal sacrifice AUTOLYSIS/CANNIBALISM: a. slightly autolytic b. partially lost c. completely lost Incidence of observations ___________ HALES FEMALES * mg VCM/kg b^-/day mg VCM/kg b.w./day 0 .,01? 0.17 777 0 0 ,,017 0.17 1.1' 0 100 100 100 50 100 100 100 100 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 1 101 10 364 26 064 25 204 1 0 skin / sUBCirris I. tumour or suspected of tumour 2. alopecia 3. 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 CLANDS 1. tumour or suspected of tumour 2. evidence of secretory activity 3. haemorrhage ABDOMINAL CAVITY 1, tumour or suspected of tumour 2. ascites 57 933 5 3 3 23 4 I 7 2 3 10 9 9 4 8 2 3 2' 1 0 1 1 1 00 00 1 00 00000 0020 1 0000 1 3 2000 2 2000 1 0000 000 1 0 01 20 1 00000 352 1 2 7 6 11 2 2 0 1 1 00 00200 00000 3 1 020 55 1 35 001 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 0 0 1 00 00000 12 1 10 3 53 20 68 526 55546 R&s 040753 UIVil/fNO STUDY NO 125 CARCINOGENICITY STUDY IN RATS WITH VCM IN THE DIET TAHLE 13. MACROSCOPIC PATHOLOGY (CONTINUED 1) Site and typo of observations SPLEEN 1. tumour or suspected of tumour 2. splenomegaly 3. atrophy A. granular surface 5. small 6. small nodule 7. dark ADRENALS 1. tumour or suspected of tumour 2. small (unilateral) 3. haemorrhagic A. 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 A. discoloured a. pale b. dark c. greenish d. reddish 5. cyst(s) 6. enlarged 7. small 8. spotted STOMACH 1. enlarged/swollen 2. haemorrhage/erosion 3. rough inner surface A. thickened cardial/fundic wall 5. thin fundic wall 6. trichober.nar Incidence of observations MALES ____________FEMALES__________ ing~VCM/kg b.w./day mgVCM/kg b.w./d.iv 0 .017 0.17 1.7 0 0~.017 0.17 1.7 0 0 10 10 8 1A 7 3 2 0 0 0 01 2 0 110 10 0 11 0 0 0 01 0 0 0 00 0 10 0 3 A 2 A3 2 0 0 0 10 0 0 1)0 0 0 20 0 0 0 00 0 0 10 0 0 7 5 2 01 1 0 1 0I 0 0 0 00 101 1 13 3 6 10 2 9 8 7 12 6 28.2,1 3 2 2 21 5 5 20 1 110 0 0 1110 0 2 0 12 1 A 2 7 A7 26 25 27 1A 23 21 9,6 A 7 1 I 5 1A 02 3 11 10 9 110 5 27 10 2 112 0 00 5 AA 3 2A 0 00 10 0 00 18 10 23 01 00 01 2A 02 00 01 0 200 5 220 3 230 3 110 0 10 0 3 3 13 0 000 I A30 112 0 0 10 0 0 000 0 0 0 0 1 0 0 1 0 0 1 A 163 1 6 5A 10 0 0 0 10 0 0 0 10 n T 1n 6 I 0 0 0 * 1A 06 00 10 00 32 12 00 00 00 0 2 0 0 0 040754 37 ffo C/3 R&S 040755 ClVO/TNO STUDY NO 125 CARCINOGENICITY STUDY IN RATS WITH VQ1 IN THE DIET TAIil i: 11. MAOt;iiSt:OI`li: PATHOLOGY (CONTINUED 2) - JO - Silo and type of observations SMALL INTESTINES 1. tumour 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. smaI1 4. filled with watery fluid COLON I. 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. edema 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 c. reddish 4. spotted 5. small haemorrhages MESENTERIC LYMPH NODES !. tumour or suspected of tumour 2. enlarged 3. small 4. edema 3. haemorrhagic 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 r.7 0 0 2 000 34 000 333 i 0 l 22 i 0 1 0 1 00 00 1 0 1 0000 0 1 00 i 2 3 1 000 0 1 000 00 0 0 0 00000 00 1 00 85 7 5 3 0 1 000 0 0 0 0 1 00000 33000 00000 00000 1 000 0 022 20 0 0 0 0. 1 0 000 0 1 0000 1 0000 0 0 .. 0 0 0 00 1 00 0 1 2 00 00400 443 1 0 00 1 00 00000 224 03 1 0000 54 1 13 00 000 00 1 00 21 1 00 2 002 1 4 030 1 1 0 1 00 01 1 00 3 0000 0 00 00 44 24 5 000 1 2 ] 2202 000 1 0 00000 00 0 0 0 1 0000 4 2 200 33 1 1 J 0 1 101 00 2 0 3 1 00 00 24 4 2 3 61 03 0 2I 21 \ 21 1 03 00 0 0 I R&s 040756 C1V0/TN0 STUDY NO 125 CARCINOGENICITY STUDY IN RATS WITH VCM IN THE DIET TARI.F. 13. MACROSCOPIC PATHOLOGY (CONTINUED 3) Site and type of 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 1. tumour or suspected of tumour 2. atrophy a. unilateral b. bilateral 3. enlarged (unilateral) 4. cryptochismus EPIDIDYMIDES 1. tumour or suspected of tumour 2. evidence of inflammation 3. atrophy 4. haemorrhage Incidence of observations MALES mg VCM/kg b-v./day 0 .017 0.17 '1.7 0 FEMALES mg VDl/kg b.w./dav 0 .017 0.17 1-.7 0 J1V0/ NO STUDY NO 125 CARCINOGENICITY STUDY IN RATS WITH VO! IN THE DIET Aiu.r. 13. iiumsroi'k: pathology (Continued 4) 56 Jili* and typo of obscrvntions JVAR1ES 1. tumour or suspected of tumour 2. cyst(s) 3. enlarged i. reddish JTERUS 1. tumour or suspected of tumour 2. hydrometra i. evidence of inflammation t, pyometra >. enlarged/swollen .IVER . tumour or suspected of tumour prominent lobular pattern swollen/enlarged discoloured a. pale b. dark . cyst(s) . $ma11 . small surface lesions (rosette, constriction) . spotted . granular surface H0RACIC CAVITY . tumour or suspected of tumour . hydrothorax . haemothorax HYMUS . tumour or suspected of tumour enlarged . spotted EART . enlarged (atria) . thrombus . pale Incidence of observations HALES mg VCM/kg b-w./day 0 .017 0.17 1.7 0 FEMALES mg VCM/kg b.w./d:iv 0 .017 0.17 1.7 0 6 4 2 35 8 II 15 3 7 5 2 4 27 2 1 I I'0 21 29 28 2113 110 0 20 10 2 2 II 33 2 00 00 27 1 1 27 I 2 I 380 1 4 200 0 13 1 0 24 5 1 0 32000 3.2 0 1 0 13I 2I 3300 1 000 1 0 2 1 5 4 0 17 14 22 33 3 0 1 1 02 00200 10101 56823 20020 3 000 1 2 1 622 0 1 200 00000 35 83 0 4874 0 00000 262 1 0 5 I 000 1 1 000 0 1 000 I 0000 00020 1 000 I 00000 14 19 16 5 3 37 650 00 I 00 675 0 I 454 00 00000 J R&S 040757 tVi)/TNO IUI)Y NO 125 CARCINOGENICITY STUDY IN RATS WITH VCM IN THE DIET Mil I 'J. MACKOKCOPl C l'ATHOEOCY (CONTINUED 5) - V) - R&S 040758 itu and type of observations IEDIASTINAL LYMPH NODES . tumour or suspected of tumour 2'. enlarged' -UNGS 1. tuiaour or suspected of tumour l. evidence of CRD 3. spongy i. spotted 5. haemorrhage 5, discoloured a. pale b, dark c. reddish 7. atelectasis (partially) 6, pleuritis TRACHEA 1. filled with blood AORTA t . 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 GLANDS 2. tumour or suspected of tumour 2, edema 3. discoloured a. dark b. reddish A. enlargcd/swollen Incidence of observations MALES tug VCM/kg b W # /day 0 .017 0.17 .7 0 FEMALES mg VCM/kg b w. /day 0 017 0.17 .7 * 3I '^~Y ~ 2 0 I 0 00 i m~ 6 "I ~ 0 ' 2 000 T" 2T 010 4 64 6 8 II 18 18 26 1 04 1 20 1 00 131 1 24 000 0I 5 72 1 50 96 01 42 00 00 00 02 000 4 3 10 554 20 17 9 00 1 564 000 020 1 03 1 00 0i 9 67 3 36 64 01 02 00 00 22 01 0 I 000 00000 599I 0 I 0000 52 I 1 2 00000 I 0000 0 00 1 0 0I I01 00000 1 0000 0 1 1 00 00000 00000 I 1 1 00 121 1 1 1 0000 00 1 00 1 1 000 0000 I 000 1 0 00000 0 1 000 00000 CRD = chronic respiratory disease C1V0/TN0 STUlJY NO 125 CARCINOGENICITY STUDY IN RATS WITH VCM IN THE DIET TAI'.I i' 13. MACKOSCOl'l i" PATHOLOGY (CONTINUED 6) - 60 - R&S 040759 Silt* and type of observations SUBLINGUAL SALIVARY GLANDS 1. tumour or suspected of tumour 2. enlarged/swollen 3. haemorrhage A. edema 5. discoloured a. pale b. reddish PAROTID SALIVARY GLANDS 1. discoloured, pale 2. spotted 3. enlarged THYROID 1. 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 A. 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 VQl/kg b.w./Jav 0 .017 0.17 1.7 0 1 0000 0 I 000 1 0000 0 1 1 00 1 0000 00000 0 0 0 0 ,0 00000 00000 00000 00000 0 I 000 3 0 0 10 0001 3 0 1 000 00000 00000 10 0 0 0 2 63 1 0 301 1 0 1 0000 21 30I 00000 1 2000 0000 1 000o0 0 'I 0 0 I 00 1 0 0 10 0 0 0 0 0 0 0 0 2 2 0 2A 0 2 111 1 6 6 2 3 2 2 10 I 3 3 3 0 02 6 9 9 2 21 8 6 6 2 0 12 12 3 7 3 13 1 13 7 9 16 2 1A 3 5 00000 0 1110 0 0 0 10 1 0C00 00000 00000 0020 00 000 iIVn/YNO STUDY NO I V> CARCINOGENICITY STUDY IN HATS WITH VCM IN THE DIET i mu r n. macijoK'tiimc pathology (CONTINUED 7) - () i Site and type of observations * PITUITARY 1. tumour or suspected of tumour 2. haemorrhagic (cyst) 3. discoloured a. pale b. dark 4. enlarged/swollen NOSE 1. tumour or suspected of tumour 2. haemorrhagic scabs around nose ORAL CAVITY 1, 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 4. haemorrhage ABDOMINAL/LUMBAL LYMPH NODES 1. tumour or suspected of tumour 2. enlarged ABDOMINAL WALL 1. small cysts TAIL 1. ulcer 2, abscess EAR DUCT 1. compact debris 2. evidence of inflammation LIMBS 1. tumour or suspected of tumour 2. abscess 3. deformed Incidence of observations MALES mg VCM/kg b*w./day 0 .017 0.17 1.7 0 FEMALES mg VCM/kg b.w./dav 0 .017 0.17 1.7 0 8 9 10 5 3 13 13 17 10 5 1 23 I 1 5 10 11 4 2 3 I 1 I 0 2 4000 I I 00 I 0000 1 1 020I 1 1 00 1 00000 6 6 2I I 0 03 0 0 4 32 00 5 3 21 1 2 1 0 0 1 18 12 10 69 6 8 13 17 0 00 0 0 0 000 0 01 01 0 00000 0000 1 00000 0 0 0 0 o' I 0 1 00 00000 0 10 00 1 1 000 2 1 000 00 I 00 0 00 0 0 00000 I 0000 2I 1 10 00000 00000 0 0I 0 0 0 I 000 00 1 00 0 00 00 00000 20000 0000 1 42 I 0I 00000 000 1 0 I 1 1 00 ..... i R&S 040760 c i Vi>/ i ;;n mi iiv MD ii") i:,\Kci:.o(ji:i:i<:n'Y study in rats with vcm in the dint ,`:i IT. "iDJCiii'i i; i'."TH01.0UY (CONTINUED 8) Incidence of observation:; :.ii. .uni i;T>i- T nli.:i i`viil inns __ MALES mp, Vffl/kj; li.w.7dny t_____~ ir.w~u:~i r~i~.~7~o I-'ICMjM.KS inn VCM/Lp. l|. /d.iv o .uTro7ri"r. r o' MISCELLANEOUS 1. emaciated 2. deformed spine 3. deformed dorsal vertebrae 15 19 17 I 30 00 I 7 0 0 48 32 22 26 14 55 0 0 10 0 0 0 0 0 0 00 R&S 040761 CIVO/TNO STUDY NO 125 CARCINOGENICITY STUDY IN RATS WITH VCM IN THE DIETS TABLE 14 TYPE AND INCIDENCE OF HISTOPATHOLOCICAL CHANCES IN THE LIVER Type of changes^ Number of animals examined Incidence of changes Males Females fflR VCM/kg b.v. /day mg VCM/kg b.u. /day 0 0.017 0.17 1.7 0 0 0.017 0.17 1.7 99 99 99 49 100 96 100 96 49 0 99 1. Foci of cellular alteration a. clear cell foci I one or a few II several 12 e 00 8 0 b. basophilic foci I one or a few II several 42 00 3 0 c. mixed cell foci 1 one or a few 01 2 d. eosinophilic foci I one or a few 11 2 All type of foci? Total ------------------------------------ 17 12 15 Number of foci" bearing animals 16 12 15 16** 3* 8* 0 2 1 30 23*** 54 00 09 00 06 00 5 19 5** 19 5 3 13*** 1 00 0 20* 26*** 29*** 4 102 0 6 4 8* 0 0 1 10*** 2 32 34 58 7 27 31* 32*** 7** 29Z0KI SSH CIVO/TNO STUDY NO 125 CARCINOGENICITY STUDY IN RATS WITH VCM IN THE DIETS TABLE 14 TYPE AND INCIDENCE OF HISTOPATHOLOGICAL CHANCES IN THE LIVER - CONTINUED 1 Type of changes 0 2. Neoplastic nodules a. one b. a few 3. Hepatocellular carcinoma 4. Angiosarcoma 0 0 0 0 5. Liver-cell polymorphism a. slight b. moderate c. severe 27 4 1 6. Prominent sinusoidal cells a. slight b. moderate/marked 1 1 7, Cysts a* one b. a feu c* many 8, Bile duct proliferation a. slight b. moderate c. severe 1 4 0 37 9 2 Incidence of changes Hales Females mg VCM/kg b.w . /day ng VCM/kg b.w. /day 0.017 0.17 1.7 0 0 0.017 0.17 1.7 1 0 0 100 1 0 0 200 0 0 0 3* 0 1 0 0 0 100 0 1 9*** 0i 1 3" 02 0 0 0 0 0 23 26 19 12** 46 41 4 7 10** 1 14 13 1 1 30 2 3 49 23 38 8 15* 15 4 9*** 3 1 1 01 4 9 2 1 103 8 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 83 11 6 0 0 01 1 2 36 18 14* 10 '2 3* 1 l4 CIVO/TNO STUDY NO 125 CARCINOGENICITY STUDY IN RATS WITH VCM IN THE DIETS TABLE I^t TYPE AND INCIDENCE OF HISTOPATHOLOGICAL CHANGES IN THE LIVER - CONTINUED 2 Type of changes __________________________________Incidence of changes________ Males Females mg VCM/kg b.w./day mg VCM/kg b.w./day 0 0.017 0.17 1.7 00 0.017 0.17 1.7 9. Cholangioflbrosls a. slight b. moderate 33 19* 20* 15 23 16 13 17 15 2 6 8* 2 1 6 2 4 0 10. Periportal fibrosis a. slight b. moderate 1 2 2. 3 5 0 0 00 2 0 0 0 20 0 0 0 11. Periportal/centrolobular infiltrates of mononuclear cells a. slight b. moderate c* severe 14 10 8 3 7 3' 2 12 1 1 1 000 0 20 0 1 1 000 0 00 12. Foci of RES-cells occasionally accompanied by a few necrotic hepatocytes a. one or a few b. several 18 15 27 5 83 5 12 39 28 42 12* . 016 3 5 0 13, Slight haematopoietic activity 2 0 0 011 0 3 0 0 2*** 0* l 0 4 0 0 25* 3 0 P910P0 SSU C1V0/TN0 STUDY NO 125 CARCINOCENICITY STUDY IN RATS WITH VCM IN THE DIETS TABLE 14 TYPE AND INCIDENCE OF HISTOPATHOLOG1CAL CHANGES IN THE LIVER - CONTINUED 3 Type of changes 14. Single cell necrosis a. slight b. noderate/severe 15. Focal necrosis a. slight b. moderate c, severe 16. Centrolobular liver-cell degeneration 17. One or a few foci of degenerated hepatocytes 18. Haemorrhagic areas 19. Peliosis-llke changes 20. Small granuloma 21. Vacuolimation of hepatocytes mainly a. periportal I slight 11 moderate 111 severe __________________________________Incidence of changes Hales Females mg VCM/kg b.v./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 0 0 1 3 4 2* 11 3* 8 2 4* 4 4 6 6 1 2 1 003 1 2 0 1 0 o - 2 0 3 0 10 1 3 3 103 2 2 0 1 0 7* 3 0 1 2 0 0 1 0 1 0 0 0 0 1 0 0 1 0 201 1 0 0 0 0 0 002 0 0 0 3 6 li* i 3 4 24 22 29 4* 0 2 5* . 0 0 3 2 60 0 1 l, 0 0 0 0 00 0 9* 2 3 3 1 0 0 0 0 0 5*** 1 0 S9Z0t>0 s?b CIVO/TNO STUDY NO 125 CARCINOGENICITY STUDY IN RATS WITH VCM IN THE DIETS TABLE 14 TYPE AND INCIDENCE OF HISTOPATHOLOCICAL CHANCES IN THE LIVER - CONTINUED 4 b. diffuse I slight 11 moderate HI severe c. centrolobular I slight Incidence of changes Kales Females mR VCH/Rr b.w./day mg VCM/kg b.u,. /day 0 0.017 0.17 1.7 0 0 0.017 0.17 1.7 1 0 1 .0 0 0 0 00 1 0 2 .0 0 2 0 0 0* 0 2 o , 0 0 1 0 00 - 0 1 o: 0 0 0 0 00 0 0 0 2 0 ^ 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 test (one tailed). i i 99/Ofro CIVO/TNO STUDY NO 125 CARCINOGENICITY STUDY IN RATS WITH VCM IN THE DIET TABLE 15 TYPE AND INCIDENCE OF TUMOURS OF THE MAMMARY CLANDS AND INCIDENCES OF ABDOMINAL MESOTHELIOMAS Incidence of tumours Males Females mg 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 0 Number of animals examined 99 99 99 99 100 98 100 96 48 98 Mammary Rlands - Mo of tumour-bearing animals 1. Adenoma a. single b. two 2. Intraductal papilloma 3. Fibroadenoma a. single b. two c. multiple 58 00 00 00 01 00 00 3 '0 0o 00 0 0 0 .0 0o 0 '0 3 91 21*** 28* 06 01 00 4 1 0 2 1 0 0 25 02 03 10** 3 1 15 2 1 21 20*' 52 00 10 14 11* 24 11 191QP0 ssa CIVO/TNO STUDY NO 125 CARCINOGENICITY STUDY IN RATS WITH VCM IN THE DIET TABLE 15 TYPE AND INCIDENCE OF TUMOURS OF THE MAMMARY GLANDS AND INCIDENCES OF ABDOMINAL MESOTHELIOMAS CONTINUED Site and type of tumours 4, Fibroma a. single b. two c. multiple 5. Adenocarcinoma a. single b. two Incidence of tumours Males Females mg VCM/kg b.w . /day m g VCM/kg b.w./day 0 0.017 0.17 1.7 0 0 0.017 0.17 1.7 0 - A6 00 00 2 0 31 A 1 22 1 0 00 0 0 0* 0 0 ..0 01 0 0 00 01 10 0 0 03 0 5 10 0 0 00 0 2 01 Abdotne r. 1. Mesothelioma 01 2 0 03 2 2 11 * P < 0.05; ** P <0.01; *** P < 0.001, according to the Fishers' exact test Cone tailed) 89Z0K) SSb R&S 040769 70 V "\ \ i tm mum * auA bti cMmi OM/rM hK nv>ici*tUiiw<MCuIUttr<I *ft*.-inINV(Mr TKHMl I tttein ur M.ti7 IMI t- I.t7 Am N FIGURE 1 7(111(1 jf* j-K x_. PIOMI I. HA# HVT vtln V rsuu wn (Mil aomt 4f ** IO 1*4 yi ?fp HI p: mi ** >]* tu 417 * Ml pi c|wo/1 MO MOM !*!. htl la * TU* ItvM Pii4P(i IP I'cCItt Mat i COlMC *<}* C9*(1 *0Vr 4' C0Mt*L t'O.Olf mitrS c< . *'* *- 1.7 4/4 f e0tl FIGURE 2 *1 ANNEX 1 - 71 - PERCENTAGE COMPOSITION OF BASAL DIET FOR RATS AND MICE soya bean oil meal fish meal 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^) vi4tamin ADEK m*ixture 3\) soya bean oil 11 7 4 36 29.7 3 3 2 0.4 0.5 0.1 0.3 3 100.0 a ) Trace elements in salt MnS04-H20 2nd, KJ FeSO^.7H20 . CoCl2.6H20 CuSOa.5H,0 NaCl ) B-vitarain mixture Thiamin_HCl Riboflavin Pyridoxin-HCl Niacin Ca-pantothenate Biotin Folic acid Vitamin B^2"PreP* Ground sucrose (0-1 *) Vitamin ADEK mixture Vit. ADj-prills (Farmix): 2250 10 vit. A, 750 10 vit. D3/g Vit. E-dry powder (Merck) 50 1 Menadion-Na-bisu lphite (Vit. k3) Wheat starch / Batch 2380 250 20 1250 20 400 45680 E 50000 Batch 25 S 30 E 100 E 125 75 1.5 5g 50 9588.5 10000 s Ba tch 25000 800 27 800 26627 g 8 S g 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 teg diet 2.5 mg 3.0 mg 10*0 mg 12.5 mg 7.5 mg 0.15 mg 0.5 mg 5.0 mg ( 5 yg B32) added to 1 kg diet 6339 IU vit. A 2112 IU vit. D3 45 mg vita E 3 mg vit* K3 i i * 30 to 0 O Nl Nl O ANNEX 2 CONTAMINANTS REGULARLY DETERMINED IN BASAL DIET 1. Contaminants In basal diet contaminant - inorganic substances_ Ung/kg). arsenic cadmium lead mercury selenium tin potassium nitrate sodium nitrite - organochlorine_compounds_(g/|59l HCB lindane (y-HCB) a-HCH 6-HCH heptachlor heptachlorepoxide a-chlordane y-chlordane aldrin dieldrin endrin methoxychlor p,p'-DDE p,p'-TDE o,p`-DDT p,p1-DDT PCBs - organoghosphorus compounds^_(mg/lcg)_ chlorpyrifos diazinon dichlorvos fenitrothion malathion mevinfos parathion sulfotep - dithiocarbamates,_in_totali_determined_as carbondisulfide_ Ong/kg). ferbam mancozeb maneb nabam thiram zineb ziram detection limit 0.01 0.005 0.05 0.005 0.01 1 25 5 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 0.04 0.3 0.01 0.01 0.01 0.01 0.01 0.01 0.01 0.01 0.5 72 tentative maximum 0.25 0.05 1 .0 0.05 0.5 25 500 10 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 0.05 0.05 0.05 0.05 0.5 0.05 0.05 0.05 1 .0 I R&S 040771 ANNEX 2 (CONT.) Contaminants in basal diet (cont.) contaminant detection limit - polycyclic aromatic_hydrocarbons_!jJgA2^ bento(b)fluoranthene bento(k)fluoranthene bento(a)pyrene fluoranthene lndeno(1,2,3-cd)pyrene . *- 0.2 0.4 0.2 1.0 0.4 nltroso compounds (pg/kg) nitrosodiethylamlne nitrosodimethylamine nitrosodlpropylamlne nitrosomethylbutylamlne nitrosomethylpropylamine nitrosomorpholine nitrosoplperidlne nitrosopyrrolidlne 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 mycotoxins (rog/kg| aflatoxin Bi miscellaneous determinations oestrogenic activity (Tiecco test) urease activity (ApH) trypsin inhibitors (mg trypsin/g food) haemagglutinins negative reaction at dilution 0.01 - a- tentative maximum 10 10 10 so 10 10 10 10 10 10 10 10 10 o.os negative 0.3 8.0 1:200 040772 33 Co 03 ANNEX 3 - 74 - R&S 040773 NUTRIENT OPPOSITION OF BASAL DIET FOR RATS Analyses of a 1000 kg batct) produced January 24, 1980 moisture 12.6 * crude protein (N x i6.25) 20.9 X crude fat 8.9 X crude fibre 3.6 X total ash 5.6 * calciun 0.89 X phosphorus 0.72 X magnesium 0.14 X potassium 0.75 X sodiun 0.28 X iron manganese copper zinc cobalt chromiun seleniin vitamin A vitamin D vitamin E carotene vitamin K3 5200 I.U./kg vitamin 1380 I.U./kg vitamin 44 mg/kg vitamin 8^ 2.0 mg/kg niacin 1.6 mg/kg pantothenic acid folic acid choline biotin vitamin 8^2 175 ppm 95 ppm 15 ppm 50 ppm 2 ppm 3 ppm 0.24 ppm 4.7 . mg/kg 5.8 mg/kg .. 12.0 mg/kg 66 mg/kg 15.5 1.25 1600 0.22 8 mg/kg mg/kg mg/kg mg/kg vg/kg isoleucine leucine lysine methionine cystine phenylalanine tyrosine threonine tryptophan g/100 g g/16 g N 10.85 4.1 1.60 7.7 1.03 5.0 1D.39 1.89 0.36 1.75 0.89 4.3 0.64 3.1 0.81 3.9 0.21 1.01 valine arginine histidine alanine aspartic acid glutamic acid glycine proline serine g/100 g 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 kg batch produced September 9, I960. moisture 11.9 r crude protein (N x 6.25) 22.2 2 crude fat 6.4 Z crude fibre 3.1 Z total ash 5.4 Z calcium 0,90 Z phosphorus 0.78 Z magnesium 0.15 Z potassium 0.82 Z sodium 0.29 Z iron manganese copper zinc cobalt chromium selenium iodine 135 ppm 95 ppm 16 ppm 55 ppm 2 ppm 2 ppm 0.40 ppm 2.0 ppm vitamin A vitamin D vitamin E carotene vitamin 7000 I.U./kg I.U./kg 55 mg/kg mg/kg 1.8 mg/kg vitamin B^ vitamin vitamin Bg niacin pantothenic acid folic acid choline biotin vitamin B^ 6.2 mg/kg 5.4 mg/kg 11.8 mg/kg 74 mg/kg 11.5 mg/kg 2.4 mg/kg 1600 mg/kg 0.20 mg/kg 16.5 /ig/kg isoleucine leucine lysine methionine cystine phenylalanine tyrosine threonine tryptophan g/100 g 0.98 1.78 1.13 0.41 0.38 0.98 0.69 0.85 0.23 g/16 g N 4.4 8.0 5.1 1.8 1.7 4.4 3.1 3.9 1.0 valine arginine histidine alanine aspartic acid glutamic acid glycine proline serine g/100 g g/16 g N 1.14 1.21 0.44 1.19 1.93 5.2 5.4 2.0 5.4 8.7 4.20 18.9 1.12 5.1 1.42 6.4 1.17 5.3 R&S 040774 4 1t - ANNEX 5 NUTRIENT COMPOSITION OF BASAL DIET FOR RATS Analyse* of a 1000 batch produced February 27, 1981 moisture crude protein (N x 6.25) crude fat crudefiber total ash calcium phosphorus magnesium potassium sodium 12.6 2 20.6 I 6.1 2 3.8 2 *5.6 2 1.05 2 0.74 2 0.13 Z 0.75 2 0.32 2 iron manganese copper zinc cobalt chromium selenium iodine - 76 - 150 90 17 45 0.6 3 0.4 1.5 ppm ppm ppm ppm ppm ppm ppm ppm vitamin A vitamin D vitamin E carotene vitamin isoleucine leucine lysine methionine cystine phenylalanine tyrosine threonine tryptophan 6600 -1} 57 1.6 I.U./kg I.U./kg mg/kg mg/kg mg/kg vitamin vitamin B^ vitamin Bg niacin pantothenic acid folic acid choline biotin vitamin Bj^ 7.5 5.7 12.1 55 14.7 5.1 1500 0.25 15.5 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 0.81 4.0 1.52 7.5 1.03 5.1 0.38 1,9 0.33 1.6 0.88 4.3 0.63 3.1 0.78 3.8 0.19 0.9 valine arginine histidine alanine aspartic acid glutamic acid glycine proline serine g/100 g g/16 g N 0.95 4.7 1.13 5.6 0.46 2.3 1.08 5.3 1.74 8.6 3.70 18.2 1.13 5.6 1.46 7.2 1.06 5.2 ^not analysed CIVO-TNO, June 1981 ' ------- i i 1 I l i 30 Co cn o o vl V/ Ul ANNEX 6 NUTRIENT COMPOSITION OF CIVO-BASAL DIET FDR RATS Analyses of a 4000 kg batch produced by Hope Farms on Sept. 9, 1981 raoisture crude protein (N x 6.25) crude fat crude fiber total ash calcium phosphorus magnesium potassium sodium 11.3 Z 20.2 6.9 3.9 Z z z j5.3 z 0.65 z 0.7 z 0.21 z 0.8 z 0.32 z iron manganese copper zinc cobalt ch romium selenium iodine 220 105 18 55 1 1 0.25 3.0 ppm ppm ppm ppm ppm ppm ppm ppm vitamin A vitamin D vitamin E carotene vitamin 5700 .1) 56 13 <3 I.U./kg I.U./kg mg/kg mg/kg mg/kg vitamin vitamin Bg vitamin niacin pantothenic acid folic acid choline biotin vitamin B^2 9.0 7 12 70 mg/kg mg/kg mg/kg mg/kg 18.6 3.3 1700 0.24 13 g/kg mg/kg mg/kg mg/kg g/kg isoleucine leucine lysine methionine cystine phenylalanine tyrosine threonine tryptophan g/100 g g/16 8 N 0.81 4.0 1.60 7.9 1.07 0.40 5.3 2.0 0.34 1.69 0.89 4.4 0.65 3.2 0.78 3.9 0.23 1.14 valine arginine histidine alanine aspartic acid glutamic acid glycine proline serine g/100 g g/16 { 1.05 5.2 1.13 5.6 0.49 2.4 1.14 5.6 1.73 8.6 3.51 17.4 1.07 5.3 1.27 6.3 1.01 5.0 ^not analysed CIVO-TNO, June 1981 aj fio 0 o -b o CD ANNEX 7 - 78 NUTRIENT COMPOSITION OF CIVO-BASAL DIET FOR RATS Analyses of a 4000 kg ba tch produced by van Eck, Cothen, on Febr. 9, 1982 moisture crude protein (N x 6.25) crude fat crude fiber total ash calcium phosphorus magnesium potassium sodium 11.3 Z 19.8 Z 6.2 Z 4.1 4Z 5.5 Z 0.95 Z 0.66 Z 0.16 Z 0.73 Z 0.32 Z iron manganepe copper zinc Cobalt chromium selenium iodine 315 95 19 65 0.5 1.0 0.3 0.9 ppm ppm ppm ppm ppm ppm ppm ppm vitamin A vitamin D vitamin E carotene vitamin K, j 8000 -X) 56 12.5 <2 I.U./kg I.U./kg mg/kg mg/kg mg/kg vitamin vitamin vitamin B^ niacin pantothenic acid folic acid choline biotin vitamin B^ 7 mg/kg 5 mg/kg 12 mg/kg 55 mg/kg 15 mg/kg 4 mg/kg 1600 mg/kg 0.21 . mg/kg 26.8 ug/kg isoleucine leucine lysine me thionine cystine phenylalanine tyrosine th reonine tryptophan g/100 g g/16 g N 0. 79 4.0 1.52 7.7 0.99 0.40 5.0 2.0 0.32 1.62 0.84 0.51 0.77 0.22 4.3 2.6 3.9 1.11 valine arginine histidine alanine aspartic acid glutamic acid glycine proline serine g/100 g 1.00 1.08 0.43 1.02 1.65 3.48 1.06 1.22 0.94 g/16 g N 5.1 5.5 2.2 5.2 8.4 17.6 5.4 6.2 4.8 not analysed CIVO-TNO, June 1982 3J C/3 O O 'si -vl R&S 040778 ANNEX 8 - 79 - CONTAMINANTS IN BASAL DIET FOR BATS AND MICE Sample of a 1000 kg batch produced on January 24, 1980 lead cadmium mercury tin arsenic selenium crgano-P-compounds organo-Cl-compounds dithiocarbamates (as CS^) 0.95 0.06 0.02 2 0.4 0.24 mg/kg mg/ kg mg/kg mg/kg mg/kg mg/kg ND ND 0.5 mg/kg aflatoxln oestrogenic activity (Tiecco test) < 5 us/kg ND benzo(a)pyrene benxo(b)fluoranthene indeno (1,2,3-c.d) pyrene benxo(k)fluoranthene fluoranthene K-nitrate Na-nltrlte n11rosodirae thylamine nitrosopyrrolidine other nltrosamines urease activity trypsin inhibitor haemagglutlnino positive dilution 2.5 4.4 2.8 2.6 12.0 pg/kg ug/kg vS/kg pg/kg pg/kg 163 mg/kg 1 mg/kg < 0.4 <2 ND 0.15 < 0.6 none pg/kg pg/kg A pH mg tryps/g sample ND - not detectable CIVO-TNO-May, 1980 ANNEX 9 CONTAMINANTS IN BASAL PIET FOR RATS CIOOO kg batch produced 2 7 February 1981) lead cadmium mercury tin arsenic selenium A organo-P-c ompound a organo-CI"Compounds PCB's dithiocarbamates af iatoxio S1ib2,g1iG2 oestrogenic activity (Tiecco teat) bentoi a) pyrene benxoCb)fluoranthene indeno (l,2,3-c,d) pyrene benioC It) fluoranthene fluoranthene K-nicrate Na-nitrite ni trosodimethyfamine ni trosopyrrolidine other nitrosaminei urease activity trypsin inhibitor haemagglucinins positive dilution negative dilution - 80 - 0-3 0.07 0.25 < 1.0 0.35 0.4 mg/kg mg/kg mg/kg mg/kg mg/kg mg/kg NO ND ND ND '< 5 Pg/kg ND 0.8 0.9 0.6 0.3 5.7 Pg/kg pg/kg Pg/kg Pg/kg P g/kg 77 mg/kg 1 mg/kg 1.2 p g/kg 1.7 P g/kg ND 0.05 < 0.6 A pH mg tryps/g sample 1 : 10 1 : 20 NO not detectable CIVO-TNO, June 1981 6^ 0 t? 0 33 Q* CO R&S 040780 CONTAMINANTS IN BASAL DIETS FOR RATS . (1000 kg batch, produced 9 September 1980) lead cadmium mercury tin arsenic selenium 0.25 mg/kg 0.045 mg/kg 0.025 nig/kg <1.0 mg/kg 0.45 mg/kg A 0.40 mg/kg organo-P-compounds ND o rgan Or C1 -c ompound a ND dithiocarbamates ND aflatoxin C^, and C2 oestrogenic activity (Tiecco test) <' 5 yg/kg ND benzo(a)pyrene benzo(b)fluoranthene indeno (l,2,3-c,d) pyrene benzo(k)fluoranthene fluoranthene 2.0 ' 2.7 2.0 1.0 16.0 yg/kg yg/kg yg/kg yg/kg Vg/kg K-nitrate Na-nitrite 80 mg/kg 1 mg/kg nit roaod ime thy 1 amine nitrosopyrrolidine other nitrosamines 0.5 1.5 ND yg/kg V g/kg urease activity trypsin inhibitor haemagglutinins positive dilution negative dilution 0.05 <0.6 none A pH mg tryps/g ND " not detectabli CIVO-TNO december 1980 ANNEX 11 CONTAMINANTS IN CIVQ BASAL DIET FOR RATS (A000 kg batch, produced by Hope Fame on September 9, 1981). lead cadmium mercury tin arsenic selenium or g ano"- P--c ompound s malathion organo--Cl"Compounds PCB's d ithiocarbaraates (as CSj) 0.9 mg/kg 0.07 mg/kg 0.005 g/kg < 1.0 mg/kg 0.45 rog/kg 0.25 mg/kg ND 0.08 ND ND < 0.5 mg/kg mg/kg >f1 atoxin Ej tB2t oestrogenic activity (Tiecco test) <5 yg/kg ND benzo(a)pyrene benzo(b)fluoranthene indeno (1,2,3-c.d) pyrene benzo(k)fluoranthene fluoranthene benzCghi) perylene K-nitrate Na-nitrite ni trosodimethy 1amine nitroaopyr rolidine other nitrosamines urease activity trypsin inhibitor haemagglutinins positive dilution negative dilution 0.4 0.7 < 0.5 0.1 0.9 0.3 yg/kg yg/kg yg/kg Ug/kg yg/kg yg/kg 83 <1 mg/kg mg/kg 0.9 1.0 ND yg/kg y g/kg 0.05 A pH 0.51 mg tryps/g 1 : 50 1 : 100 ND - not detectable CIVO-TNO, November 1981 - 3: - R&S 040781 ANNEX 12 CONTAMINANTS IN CIVO BASAL DIET FOR RATS (A000 kg batch, produced by van Eck, Cothen, on Febr. 9, lead cadmium mercury tin arsenic selenium 1.4 0.09 0.005 < t.Q 0.3 0.3 mg/kg mg/kg mg/kg mg/kg mg/kg mg/kg o rgano-F-c ompound s organo~Cl-compounds PCB's dithiocarbamates ND ND ND < 0.5 mg/kg aflatoxin Bi>B2'Gl'G2 *5 Pg/kg oestrogenic activity (Tiecco test) ND benzo(a)pyrene 0.5 benzo(b) fluoranthene 0.1 indeno (1,2,3-c.d) pyrene < 0.1 benzo(k)fluoranthene < 0.2 fluoranthene 0.2 benz(ghi) perylene < 0.3 pg/kg Pg/kg Pg/kg Pg/kg Pg/kg Pg/kg K-nitrate Na-nitrite 34 ND mg/kg ni trosodime Chylamine nitrosopyrrolidine other nitroaaraines ND 0.7 ND Pg/kg urease activity trypsin inhibitor haemagglutinins 0.00 A pH 1.4 mg tryps/g sample positive dilution none negative dilution ND * not detectable CIVO-TNO, June 19E2 - 83 040782 Bo 03 ANNEX 13 Contaminants In drinking water contaminant lnorjanic_substances__(y2/l)_ arsenic cadmium chromium copper cyanide lead mercury selenium tine potassium nitrate sodium nitrite ammonia * - organochlorine_comgounds_^vg/l^ 1,1-dichloroethane dichloromethane tetrachloroethene te trachloromethane 1,1,1-trichloroethane trichloroethene trichloromethane HCB lindane (y-HCH) e-HCS 6-HCH heptachlor heptachlorepoxide a-chlordane y-chlordane aldrin dieldrin endrin methoxychlor p,p'-DDE p,p'-TBE O,p'-DDT p,p'-DDT PCBs orqanophosphorus compounds (yg/1) chlorpyrifos diazinon dichlorvos fenltrothion malathion mevlnfos parathion sulfotep detection limit 10 1 10 1 10Q 1 1 1 1 1,000 1,000 500 - 84 - tentative maximum 50 5 50 50 100 50 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 0.06 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 0.1 0.1 0.1 0.1 0.1 0.1 0,1 R&S 040783 !I J i ! ANNEX 13 (CONT.) Contaminants in drinking vater (cant.) contaminant - polycyclic aromatic hydrocarbons^n^/l)_ benzo(b)fluoranthene benzo (JO fluoranthene t benzo(ghi)perylene benzo(a)pyrene fluoranthene lndeno(l,2,3-cd)pyrene detection limit - S3 - tentative maximum 5 200 5 200 5 200 5 200 10 200 5 200 1 NOV 1979 3) G </> O -P. o -J as c* ANNEX 14 - 86 - CONTAMINANTS IN TAP WATER (determined in period Nov.-Dec. 1979) lead cadiaiuiB mercury arsenic selenium copper chromium rinc cyanide PPb <1 <1 <5 * 2 <1 SO <1 - s 3 ammonia nitrate nitrite chloride sodium calcium potassium iron EE < 0.01 9.6 < 0.01 23 14.5 31 1.2 0.04 EEfe dichlorranothane <S BCB 1,1-dichloroethane absent' a-HCH chloroform 0.8 lindane 1,1,1-trlchloroethane 0.2 heptachlor te trachloromathane 0.5 heptachl. epoxide trichloroethene absent aldrln tetrachloroethene absent ESI dleldrin o-chlordane benro(a)pyrene < 5 y-chlordans benzo (b) fluoranthene < 5 ondrin indeno (l,2,3,-c,d) pyrene< 5 P, p1-DDE benzo (X) fluoranthene < 5 o, p *-DDT fluoranthene < 10 p,p'-TDE benzo (g,h,i) perylene < 5 p,p'-DDT 8-HCB PCB'a DDVP aevinfoa diazlnon malathion parathion chlorpyrifos sulfotep fenitrothion Ppfa < 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. l < 0. 1 0.2 R&s 040785 R&S 040786 ANNEX 15 CONTAMINANTS IN TAP WATER (determined in period Jan.-April 1980 lead cadmium mercury arsenic selenium copper chromium zinc cyanide EI& ' 1 0. 1 0.1 7 <1 55 '1 3 <2 ammonia ^nitrate nitrite chloride sodium calcium potassium iron EEE 0.01 11 < 0.01 30 )6.5 31.0 1.3 0.08 b dichloromethane 1,1-dichloroethane chloroform 1,1,1-trichloroe thane tetrachloromethane trichloroethene tetrachloroethene <5 <1 0.3 < 0.1 < 0.1 0.3 < 0.2 PPt senzo(a)pyrene <5 cenzo(b) fluoranthene <5 indeno (l,2,3,-c,d) pyre ne < 5 oenzo (k) fluoranthene <5 fluoranthene < 10 ryuizo (g,h,i) perylene <5 HCB a-HCH lindane heptachlor heptachl. epoxide aldrin dieldrin a-chlordane y-chlordane endrin p ,p * '-DDE o ,p1-DDT p,p *-TDE p,p1-DDT 6-uch 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 < 0.10 < 0.03 PCB'S < 0.8 DDVP tnevinfos diazinon malathion parathion chlorpyrifos sulfotep fer.itrothion < 0.1 < 0.1 < 0.1 < 0.2 < 0.2 < 0.1 < 0.1 < n? ANNEX 16 CONTAMINANTS IN TAP WATER (determined in period Sept.- Nov. 1980) 88 - R&S 040787 lead cadmium mercury arsenic selenium copper chromium zinc cyanide EEt <1 < 0.1 0.1 1 A <1 50 <1 10 <1 ammonia nitrate nitrite chloride sodium calcium potassium iron ppb dichloromethane 1, l-dichloroethane chloroform 1,1,1-trichloroethane tetrachloromethane trichioroethene te traehloroe thene <5 0.3 0.3 0.1 < 0.1 0.4 < 0.2 t benzo(a)pyrene . benzo(b)fluoranthene <5 <5 indeno (1,2,3 ,-c,d)pyrene < 5 benxo(k)fluoranthene <5 fluoranthene < 10 benio(gh,i)perylene <5 HCB o-HCH lindane heptachlor heptachl. epoxide aldrin dieldrin o-chlordane y-chlordane endrin p,p'-DDE o,p1"DDT P,P'-tde P,P'"DDT B-HCH methoxychlor m < 0.01 8.0 < 0.01 25. 15.0 28.5 1.2 0.01 EES: < 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 December 1980 DDVP mevinfos diazinon ma lathion parathion chlorpyrifos sulfotep fenitroth ion < 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 1980 - R&S 040788 lead cadmium mere ury arsenic selenium copper chromiurn cine cyanide b 1 < 0.1 < 0.1 2 <M 30 <1 2 6 dich1oromethane 1,1-dichloroethane chloroform 1,1,1-trichloroethane ^ <5 <1 < 0.1 < 0.1 tetrachloromethane trichloroethene tetrachloroethene < 0.05 1.0 < 0.4 benzo(a)pyrene benzo(b)fluoranthene <5 <3 indeno (1,2,3,-c,d)pyrene < 5 benzo(k)fluoranthene fluoranthene <5 <10 benzo(e,h, Operylene <5 ammonia nitrate nitrite chloride sodium calcium potassium iron E2 < 0.01 16 < 0.01 25 14.0 30.0 1.1 0.03 UCB o-HCH lindane heptachlor heptaehl. epoxide al'drin dieldrin o-chlordane T-ehlotdane endrin p,p1-DDE o.p'-DUT p.p'-tde p.p'-dut fl-HCH methoxychlor 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 < 0.1 < 0,03 /> o V AA OO >-- CO CIVO-TNO May 1981 PCB's DDVP mevinfos diazinon melathion parathion chlorpyrifos sulfotep Eenitrothion < 0.1 < 0.1 < 0.2 < 0.2 < 0.1 < 0.1 < 0.2 ANNEX 18 - 90 - CONTAMINANTS IN CIVO TAP HATER (determined in period Sept. - Nov. 1981) lead cadmiun mereury arsenic selenium copper chromium zinc cyanide ppb <1 <1 <5 < 10 <1 * 50 < 10 5 < 10 dichloromethane 1,l-dichloroethane chloroform 1,1,1-trichloroethane te trachlorome thane PPb <5 <1 < 0.1 0.2 0.2 trichloroethene te trach 1 oroe thene 1.5 < 0.5 benzo( a) pyrene El <2 benzo(b)fluoranthene <2 indeno (1,2,3,-c,d)pyrene <10 benzo(k)fluoranthene <2 f luoranthene <1 benzoCg,h,i)perylene <3 ammonia nitrate nitri te chloride sodium calcium potassium iron PPW < 0.01 9.3 < 0.01 30 17.5 30.0 1.2 0.03 HCB a-HCH lindane heptachlor heptachl. epoxide aldrin dieldrin a-ehlordane y-chlordane endriri p, p '-DDE o, p'-DDT p,p'-TDE P, P1 -DDT b-hch methoxychlor b < 0.01 < 0.01 < 0.01 < 0.01 < 0.02 < 0.02 < 0.03 < 0.03 c 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 diazinon mala thion para thion chlorpyri fos sulfotep fenitrothion < 0.1 < 0.1 < 0.1 < 0.2 < 0.2 < 0.1 < 0.1 < 0.2 R&S 040789 I ANNEX 19 CONTAMINANTS IN TAP WATER (determined in period Jan - March 1982) - 91 - R&S 040790 1 ead cadmium mercury arsenic selenium copper chromium line cyanide PPb <1 <1 <5 <10 <1 4 AO *: 10 10 ' 10 dich1oromethane PPb <5 1,1-dichloroethane <5 chloroform 1,1,1-trichloroe thane < 0.5 <0.2 te trachlorome thane < 0.1 trichloroethene te tra chioroe thene 0.A < 0-2 ppt benzo(a)pyrene < 1.3 benzo(b)fluoranthene < 0.5 indeno (1,2,3,-c,d)pyrene < 1.3 benzo(k)fluoranthene <0.7 fluoranthene benzo(g,h,i)perylene < 0.5 < 5.A CIVO-TNO May 1982 ammonia ' nitrate nitri te chloride sodium calcium potassium iron UCB o-HCH lindane heptachlor heptachl. epoxide aldrin die ldrin o-chlordane y-chlordane endrin p, p1-DDE o.p'-DDT p,p'-TDE P* P1"DDT B-HCH methoxychlor PCB's DDVP mevinfos diazinon malathion parathion chlorpyrifos sulfotep fenitrothion EES < 0.01 8.3 < 0.01 23 13.0 30.0 1.1 0.02 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 < 0.8 < 0.1 < 0.1 < 0.1 < 0.2 < 0.2 < 0.1 < 0.1 < 0.2 ANNEX 20 u' I NO - 92 OSS REFERENCE LISTING AT NC). 125 0 ZERO REFERENCE KATE. " '.'1/ Hr 79 MALES OMR 0002K 0004K 0006K 0008K 001 OD 'G012K '00140 '00160 '00181 '0020D '0022D '002AH '00260 '002BD '00300 '00320 '00340 '0036K '00380 '00400 '00421 '00440 '00460 '00480 '00500 '00520 '007.4H '0056D '0058D '00600 '00620 '00640 '0066K '00680 '00700 '00720 '00740 '0076K '0078K '00800 '00820 '00840 700860 700880 'ooyoo 700920 '00940 '00V6K .'00980 EXFR A 'j A4 A6 A8 A 10 A 12 A 14 A 16 A 18 A 20 A 99 A 24 A 26 A 2B A 30 A 32 A 34 A 36 A 38 A 40 A 42 A 44 A 46 A 48 A 50 A 52 A 54 A 56 A 58 A 60 A 62 A 64 A 66 A 68 A 70 A 77 A 74 A 76 A 78 A 80 A 82 A 84 A 86 A 88 A 90 A V7 A 94 A 96 A 98 KGR CAGE EARR (0) CO) (0) (0) <01 (0) <0) <0> (0) (0) (0) <0) <01 (0) (0) (0) (0) CO) CO) CO) CO) CO) CO) CO) CO) (0) CO) CO) CO) (0) CO) CO) <0> 0) >,0) (0) (0) CO) CO) CO) (0) CO) (0) CO) CO) CO) CO) CO) (0) 2 7. K1 7 R2 n A- LI 2 L2 4 R1L1 4 R1L2 4 R2L1 4 R2L2 4 R3 67 6 K1 6 R2 6 LI 6 L2 8 R1L1 8 R1L2 B R2L1 8 R2L2 8 R3 10 7 10 R1 10 R2 10 LI 10 L2 12 R1L1 1 2 R1I.2 12 R2L.1 12 R2L2 12 R3 14 7 14 R1 14 R2 14 LI 1 4 L.2 16 R1L1 1 6 R1 L.2 16 R2L 1 16 R2L2 1 6 K3 18 7 18 K1 18 R2 18 LI 18 L2 20 R1L1 20 k 11' 20 R2I 1 VO R2L2 FEMALES COMR EXF'KC SGR 770001H 7700031) 7700050 770007D 7700090 770011K 7700130 7700150 770017K 7700190 77002)0 7700230 770025K 770027D 7700290 7700310 770033K 7700350 770037K 7700390 77004IK 7700430 770045K 770047K 7700490 770051K 7700530 7700550 7700570 7700590 7700610 7700631 770065K 7700671 7700690 7700710 7700730 7700750 7700770 770079J 77008111 7700H3K 7700850 7700870 7700890 7700910 7700930 7700950 7 7009'0 A1 A3 A5 A7 A9 A 11 A 13 A 15 A 17 A 19 A 2.1 A 23 A 25 A 27 . A 29 A 31 A 33 A 35 A 37 A 39 A 41 A 43 A 45 A 47 A 49 A 51 A 53 A 55 A 57 A 59 A 61 A 63 A 65 A 67 A 6V . i 7.1 A 73 A 75 A 77 A 79 A HI A 83 A 85 A 87 A 89 A 91 A 93 A 95 A 9' (0) CO) CO) (0) (0) CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) (0) CO) CO) CO) CO) CO) CO) CO) CO) (0) (0) CO) (0) CO) CO) CO) CO) (0) CO) (0) CO) CO) (0) GAGE 1 1 1 1 1 3 3 3 3 3 5 5 5 5 5 7 7 7 7 7 9 9 9 9 9 11 11 11 11 11 13 13 13 13 13 15 15 15 15 15 17 17 17 17 17 19 19 19 19 EARK 7 K1 R2 LI L2 R1L1 K1L2 R21.1 R2L2 R3 7 kl R2 L. 1 L2 R1L1 R1L2 R7L1 K2L2 R3 Z R1 R2 LI L2 R1L1 K1L.2 R'.'l. .1 K2L2 R3 7 Kl R2 LI l2 R1L1 K11.2 R21.1 R2L2 R3 7. RX R2 LI l2 R1L 1 KXl 2 k:-i. i R2L2 R&S 040791 K - killed D - dead I * ill - (dead) ANNEX 20 (CONT. 1) :U/I NO - S3 1 0 S S REFERENCE LIS TING 1A> NU. 1 20 0 ZFR8 RK F F h'F NCE DAI L = 21/ k.- `V hALES COMR F..XPR 70100D /0102M 70104D 70106D 7010611 70110D '70112D '70114 D '70116D 701ian '70120D '70122D '70124D '70126D '7012811 '70130D >70132D '70134D '70136D 770138K /70140K 770142D 770144D 770146D 7 7 014 8 D 7701SOD 770152D 7701541 770156D 770J58D 77Q16GD 770162K 770164K 770166K 770168K 770170K 7V0172D 770174D 770176D //0178K /70180K 7701B2D 7 7 018 4 D 7701BAD 770168D 770190K 770192D 770194k 770196D 7701VBD 770200D A100 A102 A104 A106 A3 08 A110 A112 All 4 A116 A3 18 A120 A122 A124 A126 A3 28 A130 A132 A134 A136 A138 A140 A142 A144 A146 A148 A150 A152 A3 54 A156 A158 A160 A162 A3 64 A166 A168 A170 A3 72 AI 74 A1 76 A178 A180 A3 82 Al 84 A166 A188 A190 Al 97 A194 Al 96 Al 98 A 7 00 SGP CAGE EARR (0) <<>> (0) (0) (0) (0) (0) (0) (0) (0) (0) (0) (0) (0) to> (0) (0) (0) (0) (0) (0) (0) <0> (0) (0) <0> (0) (0) (0) (0) (0) (0) (0) (0) 10 > < <J (0) (0) (0) (0) (0) (0) (0) (0) (0) (0) (0) !0) <01 (0) 20 22 99 22 99 2? 24 24 24 24 24 26 26 26 26 26 28 28 28 28 28 30 30 30 30 30 32 32 32 32 3? 34 34 34 34 34 36 36 36 36 36 38 38 38 38 38 40 40 40 40 40 K3 Z R1 R2 U 1.2 R1L1 R1L2 R2L1 R2L.2 R3 7. R1 R2 LI 1.2 R1L1 R1L2 R21.1 R2L.2 R3 Z 1 R1 R2 1. 1 1.2 R1L1 R1L2 R21 1 R2I..2 R3 Z R1 K7 1. 1 L.2 R1L1 R1L2 R2I 3 R2I. 2 R3 Z R1 R? I. 1 1-7 R1L1 R1L2 R2L 1 R2L2 R3 FEMALES COMR t XFR SGF' CAGE ' EARR 77009911 770101D 7701030 770105H 770107D 7701OVD 7701111 7701131 770115K 770137D 7701190 7701211 7701231 770125K 770127D 770129D 770131D 770133K 770135D 770137D 770139D 77014 IK 770143D 7701451.1 770147D 770149D 770151D 7701530 7701 551.1 770157D 770159K 770161D 770163D 7701651.1 770167K 770169D 770171D 770173k 7701 751.1 7/0177D 770179D 7 70181D 770183D 770185D 770187D 770189K 770191K 770193D 7/01V5D 7 7 019 7 D 7703 99k A 99 A101 A103 A105 A107 A109 Al 11 All 3 Al 15 Al 1 7 Al 19 A121 A123 A125 A127 A129 A131 A133 A135 Al 37 A139 A3 43 A143, A145 AI 47 A149 AlSl A153 A 5.55 A157 A159 Al 61 A163 A165 A167 A169 Al 71 A173 A3 /5 A177 A179 Al 81 A183 A185 A187 A189 AI yi A 3 v;i A195 Al 9/ A >.99 <0 > <0) (0) (0) <0) <0) (0) <03 (0) < 0) (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> <03 (03 (0) <03 (03 (0) <03 <03 <03 <0 3 <03 <03 <<>) <03 19 k`3 21 7 21 R1 21 R2 21 1.1 21 t 2 23 R1L 1 23 R1L2 23 R?l 1 23 R2L2 23 R3 25 7 25 R1 25 R2 25 LI 25 1.2 27 R1L1 27 R1L2 27 R2L1 27 R21.2 27 R3 29 Z 29 . R1 29 R2 29 1.1 29 L 2 31 Rl L1 31 R1L2 31 R2L1 31 R2I. 2 31 K3 33 Z 33 Rl 33 K? 33 1. A 33 L 2 35 R1L1 35 RILL1 35 R2L1 35 r:.m. 2 35 K3 37 i 37 Rl 37 K2 37 1 1 37 i 3 V R1L 1 39 Rll 2 39 kill 1 39 R2L2 3V R5 u C w o -fe o V/ to Fo ANNEX 20 (CONT. 2) l/INU 94 OSS REFERENCE LISTING 1Y NO. 125 0 ZERO REFERENt.l 061k = 2/ O' MALES FEMALES JMR Q202I' 0204B 0206B 0208P 0210D 02120 023 40 0216 D 0218D `0220K `022211 '0224D '0226K '0228D '0230D '02320 '0234K '02360 7023BD 70240D 70242K /0244D 70246K 702480 702500 70252K 702540 702560 70258N 702600 702620 702640 702660 702680 702700 702720 70274K ` 70276K `702780 `702800 '702820 '702840 '702860 '702881 '702901' '702.920 '70294K EXPR B B4 B6 B8 B 10 H 12 B 14 B 16 Ft 18 B 20 B 22 B 24 B 26 B 28 B 30 B 32 B 34 B 36 B 38 B 40 B 42 B 44 R 46 B 48 H 50 B 52 E( 54 B 56 0 58 B 60 b 62 B 64 B 66 B 68 h 70 B 72 B 74 B 76 R 78 B 80 B 82 B 84 B 86 B HH B 90 B 92 B 94 SGP CARE CO) < 0) (0) (0) CO) CO) (0) (0) <0> 10) (0) (0) <0> (0) (0) (0) (0) (0) <<>) CO) (0) (0) CO) CO) (0) CO) (0) CO) CO) (0) CO) CO) CO) CO) (0) (0) (0) CO) CO) (0) (0) CO) CO) CO) CO) CO) CO) 42 42 42 42 42 44 44 44 44 44 46 46 46 46 46 48 48 48 48 48 50 50 50 50 50 5? 52 52 52 52 54 54 54 54 54 56 56 56 56 56 58 58 58 58 58 60 60 EARN 4' COMR 7. 77020IK Kt 770203K K2 7702050 LI 7702070 L2 7702090 R1L1 7702110 R1I.2 7702130 R2L1 77021SD R2L2 770217K R3 7702190 Z 7702210 kl 7702231 R2 7702250 LI 7702270 L2 7702290 R1L.1 7702310 R1I..2 7702330 R2L1 7702350 R2L2 7702370 R3 770239K Z 7702410 R1 7702430 R2 > 7702450 LI 770247K 1.2 7702490 R1U 7702510 Rll. 2 7702530 R2L1 7702550 R2L2 770257K R3 7702590 7 770261K Kl 770263K R2 7702650 l.l 770267K 1.2 7702690 Rll-1 77027IK Rll. 2 R2L1 7702730 770275K R2L.2 7702770 h`3 7702790 Z 7702010 Kl 7702830 K2 7702850 L 1 770287K 1.2 7702890 Rll. J 7702910 Rll? 7702930 EXPR B1 B3 B5 B7 B9 B 11 B 13 B 15 B 17 B 19 B 21 B 23 B 25 B 27 B 29 B 31 B 33 R 35 1# 37 B 39 B 41 B 43 R 45 B 47 B 49 B 51 B 53 B 55 B 57 B 59 B 61 B 63 B 65 B 67 B 69 B 71 B 73 B 75 B 77 B 79 B 81 B 83 B 85 B 87 B 89 B 91 B 93 SUP LAGF . HARK CO) (0) CO) CO) (0) CO) CO) CO) CO) CO) (0) (0) CO) CO) CO) CO) CO) CO) CO) CO) CO) (0) CO) CO) (0) CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) 41 Z 4 1 Kl 41 K2 41 LI 41 L2 43 Rll.l 43 Rll. 2 43 K2I. 1 43 K2L2 43 R3 45 Z 45 Kl 45 R2 45 LI 45 L 2 47 Kill 47 R1L2 47 R2L1 47 R2L2 47 R3 49 7 49 " Ri 49 R2 49 "'Ll 49 L 2 51 K J L.l 51 Rll, 2 51 K2L1 51 R2L2 51 R3 53 7. 53 Kl 53 R2 53 L 1 53 1.2 J h) Kll. I 1") 5 Rll. 2 55 R2L1 55 R2L2 55 R3 57 7 5/ Kl 57 K2 57 1 1 57 1. 2 5V Kill ViV RJI 7 R&S 040793 ANNEX 20 (CONT. 3) /TNG 95 - OSS REF E R E N C E LI STING .Y NO. 125 0 21. Nil Kl l K Kh Nl.t BAIL 1 HALES JMR E XPft 02960 02980 03000 03020 03040 03060 0308K 03100 03120 03140 '03161 '03180 '03200 '03220 '03240 ?03260 /0328D 703300 703320 70334D 703360 703380 703401 703420 703440 703460 703480 70350K 703520 70354K 703560 703580 '70360K '703620 70364K 703660 '703680 '703700 '70372K '703740 '703760 ?70378k 770380K 7703820 7703840 7703860 7703880 770390I 7703920 7703940 770397,0 770398K 7704000 B 96 B 98 B100 0102 0104 0106 0108 B110 0112 B114 0116 0118 0120 0122 0124 0126 0128 0130 0132 0134 0136 0138 0140 0142 0144 0146 0148 0150 015? 0154 0156 0158 0160 0162 0164 0166 0168 0170 HI 72 0174 0176 0178 0180 0182 0184 0186 0188 0190 0192 0194 0196 0198 0200 SUP CAGE EARR (0) 60 R2L1 (0) 60 K2L2 (0) (0) (0) (0) 7,0 62 62 62 K3 Z R1 K2 (0) 62 1.1 (0) 62 L.2 <01 64 R1L1 (0) 64 R1L2 (0) 64 R2I 1 <0) 64 K?l. 2 (0) 64 (0) 66 K3 Z (0) 66 Kl (01 66 R2 (0) 66 1.1 <01 66 L.2 (0) 68 R1L1 <01 68 R1L2 (0) 66 K2-I.1 (0) 68 R2L2 <01 68 R3 (0) 70 Z (01 70 R1 (0) 70 k? (01 70 l1 (01 70 L2 (01 72 R1L1 (0) 7? K1L2 <01 7.V r;/i 1 (0) 7? K2I.2 <01 T.; R3 (01 74 Z (01 74 Kl (0) 74 u:.' (01 74 ii (01 74 L2 (01 76 R1L1 (01 76 KU 2 (01 76 kl.'l. .1 (01 76 K2L 2 (01 76 R3 (01 78 Z <01 7H Kl (0) 7H (0) 76 (0) 76 K2 11 L2 (01 80 K1L 1 (0) lit* KU.2 <01 60 K3L 1 (0) 80 K21.2 (0) HO K.l E H MAt E S COMR EXFK 6GF CAGE EARK 7702V5K 0 95 (0) 59 R21.1 7702970 B 97 (0) 59 K2I 7 7702990 B 99 (0) 5V R3 7703010 01 01 (0) 61 2 7703031 B103 (0) 61 R1 770305K BIOS (0) 61 K? 7703070 B107 (0) 61 1. 1 770309K B109 (0) 61 1.2 7703110 Bill (0) 63 R1L1 770313.1 0113 <01 63 R1L2 7703150 BUS (0) 63 K21.1 7703170 B117 (0) 63 R2I..2 7703190 B119 (0) 63 770321.0 0121 (0) 65 K3 1 7703231 BUM (0) 65 Rl 7703250 B123 <0) 65 K2 770327K B127 (0) 65 1. 1 7703290 B129 (0) 65 L2 7703310 B131 (0) 67 R1L1 7703330 B133 (0) 67 R1L2 7703350 B135 (0) 67 R21 1 7703370 B137 <01 67 R2L2 7703391 B139 to > 67 R3 7703410 B141 <()) 69 2 7703430 B143 (0) 69 Rl 7703450 B145 (01 69 K7 770347K B147 (0) 69 1. 1 7703490 B149 <(>) 69 L2 770351K 0151 <01 71 R1L1 770353K B.153 (0) 71 r 11:1 7703550 B155 (0) 71 k:.'.i. 7703570 B157 (0) 71 k:.>i.:- 7703590 B159 <<)> 71 Rj 7703610 B161 tO) 73 Z 7703631 0163 <0> 73 Rl 770365K B165 (0) 73 K? 7703670 B167 <0) 73 1( 770369k B16V (0) 73 L2 7703710 :l 7 i 10) 75 RlL 1 7703730 0173 <0> 75 Kl L 2 7703750 B175 (0) 75 k:.m. i 7703770 B177 (0) 75 k:.'i. .' 7703790 HI 79 10} 75 K3 7703810 BiBt (0) 77 7 V /(13830 0183 (0) 77 kt 7703850 B185 <0> 77 k 770387K 0187 (0) 77 11 770389K B i 89 (0) 77 L3 7703910 B > y .1 (0) 79 Kill 7703930 0193 (0) 7 V KH . 7703950 0 1 95 (0) 79 K.'l 1 7703970 0 ,i 9 7 <0> 7V R..'i 7 770399i 0 i 99 (0) 7" K 040794 oo Bo </> ANNEX 20 (CONT. 4) 0/1 NU OSS REFERENCE LI STING - 96 * ay nu. 175 0 7LR0 Rb.FERt.Nt> HA lb - 7.* MALES :omr '04020 70404K 704060 704080 704 ion 70412K 70414K 70416K 70418K 704200 704220 704240 704260 704280 704300 704321* 704340 704360 70438D 704400 704421* 70444D 704460 704480 70450K 70452D 70454D 704560 704380 704600 7Q462K 704640 '70466K '70468K '70470K '704720 70474K '704760 '704780 '704800 '704Q20 '704840 7704860 /704880 '704900 EXF'R C2 C4 C6 C8 C: 10 C 12 C 14 i; 16 c 18 c 20 c 22 c 24 t; 26 c 28 c 30 c 32 c 34 u 36 (; 38 c 40 c 42 L 44 t: 46 t: 48 c 30 c 52 c 54 c 36 i: 5B c 60 c 62 L 64 t; 66 t: 68 c 70 c 72 t. 74 t; 76 c. 78 c 80 c 82 c H4 t: H6 c 88 c 90 SGR CAGE FQKK (0) 82 7. CO) 82 R1 (0) 82 R2 CO) 82 L1 CO) 82 L.2 CO) 84 Kll. 1 CO) 84 K1L2 CO) 84 R2L 1 CO) 84 R2L2 <0) 84 R3 CO) 86 2 CO) 86 R1 CO) 86 R2 (0) 86 LI <0) 86 1.2 CO) 88 Rll. 1 CO) 88 R1L2 CO) 88 R2L1 CO) 88 R2L2 CO) CO) 88 90 K3 Z CO) 90 R1 CO) 90 R2 CO) 90 LI CO) 90 1.2 CO) 92 R1L 1 CO) 97 R1L2 CO) 92 R2L1 CO) 92 R2L.2 CO) 92 R3 CO) 94 7 CO) y4 R1 CO) 94 R2 CO) 94 1.1 CO) 94 1.2 CO) 96 RIL. 1 CO) 96 R1L2 CO) 96 R2L1 CO) 96 K2I. 7 CO) CO) 96 98 R3 7 CO) CO) 98 98 K1 R2 CO) 98 11 CO) yH 17 FEMALtS C8MR EXF'R 7704G1K 7704030 7704050 7704070 77040VK 7704110 7704130 770415K 7704170 7704190 7704210 7704230 7704251 7704270 7704290 770431K 7704330 7704321 770437K 770439K 7704410 7704430 7704450 7704470 770449K 7704510 7704530 7704330 7704570 7704390 770461K 770463K 7704631 770467K 7704690 7704710 7704730 7/0475K 7704770 7704790 7704810 7704830 7704H3K 7704870 7704890 C1 (.: 3 03 C7 89 C 11 C. 13 8 15 r: 17 0 19 C 21 c: 23 c; 73 <; 27 C 29 C 31 C- 33 C 33 C 37 C 39 C 41 C 43 C 45 C 47 C 49 C 51 L 33 0 55 C 57 C 59 C 61 C 63 C 63 U 67 C 69 C 71 <: 73 C 73 (J 77 C 79 C 81 I) 83 C 83 c; 87 C 89 SGP CO) (0) CO) CO) CO) CO) CO) (0) CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) (0) CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) (0) (0) CO ) CO) CO) CO) CO) CO) LAO. h ARK 81 / HI R1 81 R2 81 L 1 81 1 7 83 Kll. 1 83 K1L2 83 R2L1 83 R2l 2 83 K3 83 7 83 R1 85 R2 85 L 1 83 1.7 87 Kll. 1 87 R1L2 87 R7L1 87 R7.L7 87 K3 8V 7 89 R1 89 R2 89 L 1 89 I..2 91 Kll. 1 91 R1L2 91 R2.L 1 91 K2I.7 91 K3 93 7 93 R1 93 R2 93 1.1 93 1.7 93 K1L1 93 R1L2 95 R?l 1 93 K7I 3 93 K3 97 7 97 ra 97 K.' 97 1 9/ u Qo CO O C* 0 tXoI 01 Al'NEX 20 (CONT. 5) O/TNO OSS REF E R N C E L AY NCI. 125 0 MAL.F.S OMR FXPR SOP CACiF EARR 0492D '0'19'Ui '0496K '04981 '0500K '05021 '0504D '0506K '050BD '0510D '0512D '0514n '0516D '0518D '0520D '0522D '0524D '0526D '0528D '0530K 70532D '0534D '0536D '0538D '0540D '054211 '0544K '0546D '0548D '0550D '0552K '0554D '0556D '0558D '0560D '0562D '0564D '0566D '05681 '0570D '0572I'i '0574D '0576D '0578D '05B0D '0582K '0584D '05861 '03888 '0590D '0592D '0594 L1 '0596D ' 0 5 9 0 [ '0600H i: 92 C 94 C 96 C 98 C100 0102 Cl 04 C106 C108 Cl 10 Cl 12 C114 C116 Cl 18 Cl 20 Cl 22 C124 C126 Cl 28 C130 Cl 32 C134 C136 C138 C140 C142 C144 C146 C1 4 R Cl 50 Cl 52 C154 C156 Cl 58 Cl 60 Cl 62 C164 C166 Cl 68 C17 0 ri72 C174 C176 Cl 78 C180 Cl 82 C184 C186 1.188 C. 190 Cl 9 2 ci y4 Cl 96 Cl 98 C200 (0) (0) (0) (0) <01 (0) (0) <0) <<)> CO) (0) (0) CO) CO) CO) CO) CO) CO) CO) (0) CO) CO) CO) CO) CO) CO) CO) (0) CO) CO) CO) CO) CO) CO) (0) CO) CO) CO) CO) (0) CO) CO) CO) CO) CO) CO) CO ) CO ) CO) CO) CO) CO) (0) CO) CO ) 100 100 100 100 100 102 102 1 02 102 102 104 104 104 104 104 106 106 106 106 106 108 108 108 108 108 110 1 10 110 110 110 112 1 17 ;i y 11 112 1 14 114 i 14 114 114 1 16 11A i 16 116 116 1 18 1 18 l iH 1 18 1 1 II l 70 120 120 1 20 i 70 Rll 1 nil:? K21.1 K2L2 R3 7. R1 K2 LI L2 Rll.l Rll. 2 R21.1 R2L2 R3 7. Ri R2 LI L2 R1L1 R1I.2 R2I.1 R2L2 R3 7. R1 R2 LI L2 KI LI K1I..2 R2I 1 R2L2 R3 7 Ni R2 LI L2 R1I.1 Rll. 7 K2L1 R2L2 R3 7. IU R2 11 i R1L1 R1 L2 R2I 1 K7I 7 K3 - 97 - TING 7H RCJ RfcPFR'ENCL. DA It - 217 PF. HALF'S COMR F.XPR SCiP tlAGF EARR 770491D 7704931! 770495K 770497K 770499K 7 705011.1 770503K 770505D 770507K 77O509D 77051IK 77051 3D 770515D 770517K 770519D 77052ID 770523K 770525K 770527H 7705291 7705311.1 770533IJ 770533D 770537D 770539D 77054ID 770543K 7705451 770547D 770549D 77055ID 7705530 77055SD 770557D 7705591 77056ID 7705630 770565D 770567D 770569D 77057.1 D 7/05731' 770575D 770577D 77057911 770501D 7/05838 770585K 770587D 7705898 770591K 7 7 0 5 y 311 7 705 95)! //059 71s 77059911 0 91 C 93 C 95 C 97 C 99 ClOl Cl 03 C105 Cl 07 C109 cm C113 C115 Cl 17 Cl 19 C121 C123 C125 C127 Cl 29 C131 C133 C135 C137 Cl 39 C141 C143 Cl 45 Cl 47 Cl. 4 9 cm C153 C150 Cl 57 Cl 59 C161 C163 Cl 65 Cl 67 Cl 69 C171 C1 73 Cl 75 Cl 77 Cl 79 Cl 81 C183 C1R5 C1H'7 C189 ia9i 1, 1 V/ l i v:# C1 97 C \ 99 CO) CO) CO) CO) CO) (0) CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) (0) (0) CO) (0) CO) CO) (0) CO) (0) CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) (\ \ 99 99 9V 99 99 101 101 101 101 101 103 103 103 103 103 105 105 105 105 105 107 107 107 107 107 109 109 109 109 109 111 1J1 111 111 111 113 I13 113 113 113 1J 5 1 13 1 i3 1 13 115 1 17 1.1 7 I1/ 1 17 11/ 1 tV 11 I )9 ] 1V 1 1 <2 ' K1L1 Kil. 2 K2I/1 R2L2 R3 7 kl R2 LI L2 R1L1 Nil. 2 K2I..1 K2L2 R3 7 RI R2 LI L2 R1L.J Rll-2 K'21.1 R2L2 R3 7 kl R2 LI L2 R .1 LI Nil. 2 R.VI 1 R2L2 R3 / R2 LI l .7 k :i L l Nil 2 N2L1 R2L2 R3 / l\ 1 K2 LI 12 RI L 1 K1L.2 k.'i i k;*i :* iv < ANNEX 20 (CONT. 6) J/TNO OSS REFERENCE LISTING AY NO. 125 n ZLFcO KFEFkhNCF OlYfl - 21/ 8/ 'v MALE'S FEMALES OMR 06020 0604 0 06060 06080 061 on 06121' 06140 06160 0618K '06200 06220 '06240 '06260 '0628K '06300 '06320 '06340 '06360 '06380 '06400 '06420 '0644K '06460 '06480 '06500 '0652K '06540 '06560 '06580 '06600 .'06620 70664K '06660 70668K 70670k 706720 70674K 706760 706780 706800 706820 706840 706861 706880 706900 706920 706940 EX Ok 0 '# 04 06 08 0 10 0 12 0 14 0 16 0 18 0 20 0 } rt 0 24 0 26 0 28 0 30 0 32 0 34 0 36 0 38 0 40 0 42 0 44 0 46 0 48 0 50 0 52 0 54 0 56 0 58 0 60 0 62 0 64 0 66 0 68 0 70 0 /2 0 74 0 76 0 78 0 HO 0 8? LI B4 0 86 0 88 0 90 0 92 0 94 SGF' CAGF E^kk (0; (0) <0> <0) (0) <0> <0> <0> <0> <0> <0) (0) (0) (0) (0) <0> <0> (0) (0) (0) (0) <0) (0) (0) <(>) (0) (0) (0) (0) <01 <0) (0) (0 <\ (0) (0) lO) (0) (0) (0) (OF (0) (0) (0) (0) LO) (0) 122 122 122 3r,r> 122 124 124 124 3 74 124 126 126 126 126 126 128 128 128 128 128 130 130 130 130 130 132 132 13? 132 132 134 134 134 134 ' l 34 136 136 3 36 136 136 13e 3 38 3 3H 138 1 38 3 40 140 Z R1 K2 1.1 1.2 R1L1 R1L2 R2L.1 R2L2 R3 Z R3 k2 1.1 L2 R1L1 R1L2 K2L1 R2I.2 R3 Z R1 R2 LI L2 R1L1 R3L2 K21.1 R2L 2 R3 Z K .1 R2 11 L2 R1L1 R3L 2 K2I l K21 2 R3 Z K1 R2 LI 1,2 ku l Kit 2 CUMk 7706010 7706030 7706050 7707.070 770609K 7706110 7706330 7706350 7706171 7706190 7706210 7706230 /70625k 7706270 7706290 7706310 7706330 7706350 7706370 7706390 7706410 7706430 7706450 7706470 7706490 7706510 7706530 7706550 7706570 7706590 7706613' 770663k 7706651 7/06670 '0669K 770*710 7706730 7706730 7706771 7706790 7706810 7706830 7706830 7706870 7706890 7/069.1 0 7706930 F XPR 01 03 03 07 09 0 33 0 13 0 13 0 17 0 19 0 21 0 23 0 25 0 27 D 29 0 31 0 33 0 35 0 37 D 39 H 41 0 43 0 45 0 47 0 49 0 3i. 0 53 0 55 0 57 0 39 0 6 i. 0 63 0 65 0 67 0 69 0 71 0 73 0 73 0 77 0 79 0 8i 0 H3 D 85 0 87 0 R9 0 91 0 93 SGF' (0) (0) 10) (0) (0) (0) (0) (0) (0) (0) (0) (0) (0) (0> (0) (0) (0) (0) (0) (0) (0) (0) (0) (0) (0) (0) (0) (0) (0) (0) (0) (0) (0) (0) (0) (0) (0) (0) (0) (0) (0) (0) L0) (0) (0) (0) (0) CAGE 123 121 121 121 12l 123 123 123 3 23 123 125 125 125 125 123 127 127 127 127 3 27 129 129 129 129 129 131 131 1 3.1 131 131 133 133 133 133 133 135 135 1 33 133 i 33 137 137 1 37 137 1 37 1 39 13V EAkK 4 RJ K2 1. j 12 R1L1 R1L2 k'?l 1 k'2l. 2 R3 Z kl K? 1.1 L2 R1L1 R1L2 K21.1 k2l. 2 R3 Z R3 R2 1. 1 L2 R1L1 R1L2 K2I. 1 k2l. 2 R3 Z kl 1(2 1.1. L2 1 Ll k1 L2 K2I i r:m-? R3 7 Kl K2 Ll 12 kll .' klL2 ANNEX 20 (CONT. 7) .1/1 N(J OSS REF ERE N C E 1Y NO * 122. 0 - 99 LI STING 7H k'U REFERENCE DATE 21 / 8/ /v MAI. E S EE MOLES OMR EXF'R BW cage EARR COMR t y.F-R Sbk CAGE EARR 0696D 069SH 0700D 0702K 07Q4K 0706K 0708k 0710K 0712K 0714K 0716K 0718K '0720K '0722K `Q724K '0726K '0728K '0730K '0732K '0734K '0736K '0738K '0740K '0742K 'Q744K '0746K 70748K 70750K 70752K '0754K 70756K 70758K /0760K 70762k 70764K 70766K 707 68K 70770K 70772K 707 74 K 70776K 7077BK 70780K 707U2K 70784k 70786k 70788K 707V0K /0/V2K 70794k 707V6K 707V8K 7080OK H 96 H VB IHOO D102 C104 ruoA in on 1)110 D112 0114 ) 111A mis ni2o HI 22 D3 24 HI'/A iiii'a D130 D132 D134 Li 113 A Ml 38 Ml 40 M142 [1144 IU 4 A [1148 [1150 [H52 Ii 154 H.156 1U58 [U60 [1162 HI 64 ).i 166 H168 [i] 70 [117? HI 74 HI 76 111 78 M180 HI 87 IU 84 [1186 HI 88 HI VO H 1 77 111V 4 HI V6 Hi VB H.'OO <<>) (01 (0) (0) (0) (0) (0) (0) (0) (0) (0) (0) <0> (0) <01 <<>) (0) <0> (0) (0) (0) (0) <01 <0) (0) (0) (0) (0) (0) (0) (0) (0) (0) (0) (0) (0) (0) (0) (0) ((1) (01 (01 (0) (0) ((>> (01 (0) (0) (0) (0) (0) (0) <01 140 140 140 14? 142, 142 142 14? 144 144 144 144 144 146 146 146 146 146 148 14H 148 148 148 150 150 150 150 150 152 15? 152 152 15? 154 154 154 154 154 .1 56 156 156 156 156 X58 158 158 158 158 1.60 160 1 60 160 160 R2L1 R2L2 R3 7 R1 R2 LI L.7 Rll. 1 R1L2 R2L1 R2L2 R3 7 R1 R2 11 1.2 Rll. 1 R1L2 R2L1 R21 2 R3 7 R1 R2 11 1.2 K).l 1 R1L2 R2L1 k 71, 7 K3 7 R1 R2 1. 1. 12 K1L1 R1L2 R?l 1 R2I.2 R3 Z ftl K7 LI L2 Rll 1 k (1 7 K7l 1 R7L7 R'3 770695H H 95 770697J' H 97 7706VV).i H 99 770701k D101 770703K D103 770705K [U05 770707K Ml 07 770709K D109 77071IK Dill 770713K D113 770715K HI 15 770717K HI 17 77071VK ' H119 77072IK D121 770723K H123 770725K HI 25 770727K H127 77072VK M129 770731K D131 770733K D133 770735K Hi 35 770737K Ml 37 77073VK D139 77074 IK D141 770743K M43 770745K HI 45 770747K Mi 47 77074VK D149 770751k D151 770753K III 53 770755K Ml 55 7/075 7I\ HI 57 770759k [1159 770761K D161 770763K H163 7707651 HI 65 770767k D1.67 7707A9K D169 770771k HI 71 770773k IJl 73 7707/5K 11175 770777k HI 77 //0779K D179 770781K U181 770783k 770785K Ml S3 D185 770787k Ii J 87 770789K l.iiBV 7 70/911\ H ' 91 /70/V3K D19 3 770795k HI 95 7707V7K H 197 770799K HI yy (0) (0) (0) (0) (0) (0) (0) (0) (0) (0) (0) (0) (0) (0) (0) (0) (0) (0) (0) (0) (0) JO) (0) (0) (0) (0) (0) (0) (0) (0) (0) (0> (0) (0) (0) (0) (0) (0) (0) (0) (0) (0) (0) (0) (0) (0) (0) (0) (0) (0) (0) (0) (0) 139 139 139 141 141 141 141 141 143 143 143 143 143 145 145 145 145 145 147 147 147 147 147 14V 149 149 149 149 151 151 151 151 1 51 153 153 153 153 1 53 155 1 55 155 155 155 157 157 .1 57 157 157 159 159 159 15V 159 R2L1 R2L*2 k'3 7 R1 R2 LI L2 R11.1 R1L2 F:2L1 R2L7 R3 / R1 R2 L1 I..2 R1U R1L2 R2L1 R2.L2 R3 l R1 R2 1.1 1.2 Rll.l R1L2 R2L1 k?l. 2 R3 I R1 R2 1. 1 12 K J LI R1L2 R2L1 K2I 2 K'3 1 R1 R2 LI L2 R1 L 1 Idl 2 K2I 1 R2L2 R3 i I R&S 040798 annex 20 (CONT. 8) 1/1 wo OSS REF E R E N C E LI STING - 100 - tV NO. 3 25 0 7FRCI RF.FFRFNCF HAIL - 2 3/ 8/ - L MALES FEMALES Jf(R 080 2D 08040 0806D 08080 08100 08120 0B14D 081 AD 08180 '0820D '08220 '03240 '082A0 '03280 '08300 '08320 708340 '083AD 708380 708400 70842D 708440 708460 708480 708500 708520 70854D 7-08560 708580 708600 708620 708640 708660 708680 708700 708720 708740 708760 708780 '708800 '708820 '708840 '708860 708880 '708900 '708920 '708940 EXF'R E2 E4 EA F. 8 E 10 E 12 E 14 E 16 E 18 E 20 E 22 E 24 E 26 F 28 E 30 E 32 E 34 E 36 E 38 E 40 E 42 E 44 E 46 E 48 E 50 E 52 E 54 E 56 E 58 E 60 E 67 t: 64 E. 66 t 68 E 70 E 72 F 74 F 76 F 78 F 80 F 82 F 84 t: 86 F 88 F 90 F V? F 94 SGF' CAGE fc ARK <03 <03 <03 <0 3 (03 <03 <03 <03 <03 <03 <03 <03 <0> <0 3 <03 <03 <03 <03 <03 <03 <03 <03 <03 <03 <03 <03 <03 <03 <03 <03 <03 <03 <03 <03 <03 <0 <0 ' <0 3 <03 <03 <03 <03 <<3 3 (0) <03 <0 3 <03 3 62 3 62 162 162 162 3 64 164 164 164 164 166 166 166 166 166 168 160 168 168 168 170 170 170 170 3 70 172 172 3 72 372 372 174 174 3 74 3 74 374 176 376 176 3 76 3 76 178 3 78 178 178 178 3 80 3 80 7 R1 R2 LI L2 Rll.l R 11.2 R2L1 R2L2 R3 7. Rl R2 LI L2 RILl R1L2 R2L1 R2L2 R3 1 Rl R2 LI L2 Rll.l R1L2 R2L1 R2L2 R3 7 Rl R2 L1 L.2 R1L.3 R1L 2 R2L1 R'.'L 2 R3 7. kl R7 L1 l7 R1L3 R1I.2 I.OMR 7708010 7708030 7708050 7708070 7708090 7708110 7708130 7708150 7708170 7708190 7708210 7708230 7708250 7708270 7708290 7708310 7708330 7708350 7708370 7708390 7708410 7708430 7708450 7708470 7708490 7708510 7708530 7708550 7708570 7708590 7708610 7708630 7708650 7708670 7708690 7708710 7708730 7708750 7708770 7708790 7708810 7/0HH3D 7708850 7708870 7708890 7708910 770HV30 EXF'R E1 E3 E5 E7 E9 E 13 E 13 E IS E 17 E 19 E 21 E 23 E 25 F, 27 E 29 E 31 E 33 E 35 37 E 39 E 41 E 43 E 45 E 47 E 49 F 51 E 53 F, 55 E 57 F 59 F 61 F. 63 F 65 F 67 E 69 t 73 F 73 F 75 F 77 F 7V F. 81 F 83 t 85 F 87 F 89 i- 91 F 93 SGF' CAGF . F ARK <0> <0> <0) <<>> <0) <0> <0> <0) <0> <0) <0> <0> <<>) <0> <0> <0 > to > <0) <0> <0 > CO) <0) C0> <0 > <0> <0) <0> <0> (0) <0> <0> <0) <0) <0) CO) <0) (0) <0) <0) <0) <0) <0) <0) <<)) <0> (0) <0) 161 363 161 161 161 163 163 163 163 163 165 165 165 165 165 167 167 3 67 167 167 169 169 169 169 169 171 373 171 171 371 1 73 3 73 1 73 173 3 73 3 75 3 75 3 75 175 3 75 17/ 1 77 3 77 177 1 77 1 /V 3 79 7 F('J k? LI L2 RILl R1L2 R2L1 R2L2 R3 7 Rl R2 LI L2 RILl R11. 2 R2L1 R21.2 R3 Z Rl R2 LI L2 RILl R1L2 R2L1 R2L? R3 7. R3 K7 13 L.7 Rll.l R1L2 R2LJ R2L.2 K.S / K1 R7 L1 1.7 kill K1L 7 X CO Jov o >/ CO CO ANNEX 20 fCOKT. 9) 1 HO ; s s REF E R E N C E Nil. i:".-. ci 101 LI STING ZERO REFERENCE I'Alk - 21/ 8/7 R&S 040800 MALES -IR EXPR 396D 398D 900D 902D 904D 906D 908D 9100 93 2H 914D 9160 9181 920D >92,20 >9240 >9261 >9281 >9301 >9320 >9340 19360 >9381 19401 19420 19440 19460 19480 19500 19521 19540 09560 19580 09600 09620 09640 09660 09680 09700 09720 09 74 0 09761 0978H 09801 09820 09840 09860 '09880 09901 '09920 '09940 "09960 '09980 -l 0000 E 96 F 98 H 1 00 t.102 104 E 1 06 MOB f.110 El 12 El 14 El 16 F. 118 El 20 E.l 22 E124 E126 11 28 El 30 F.132 E134 E136 E 138 E 1 40 F.142 E144 E 1 46 fc 1 48 El 50 E152 E.154 E 156 El 58 E 160 FI 62 E.l 64 El 66 E 168 E170 E172 E 1 74 E 1 76 E, 1 78 E180 k 1 8? k 1 84 E186 E 188 E 190 E 1 92 k 1 94 f 196 k' 1 98 k 200 SGP CAGE EARR <0> (0) (0) (0) <0> (0) (0) (0) <0) CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) (0 ) C01 1HC) 180 180 1 82 182 182 182 182 184 184 184 184 184 186 186 186 186 186 1 88 188 188 188 188 iyo 190 1 90 1 90 1 90 192 192 19? 192 I 92 194 1V4 194 194 1 94 1 96 1 96 196 1 96 .1 Vr. 198 1 ' 1 93 1 98 .1 98 200 200 .'00 200 200 K2L1 R2U? 83 7. K1 R2 LI L2 R1L1 K1L.2 R2L1 R2L2 R3 7. R1 R2 l.l 1.2 R1L1 FC11.2 R2L1 R2L2 R3 7. R1 R2 L1 I..2 Kit. J R1L2 R2L1 R2L2 R3 l R1 R2 11 L2 Kll 1 R1L2 R2L1 R2L 2 K3 7 K1 R? LI 1.2 RtL 1 Kll 2 K2L 1 K2L2 R3 EE'MALES COMR EXE-R SOP CAGE FARR 7708950 7708978 7708998 77090ID 7709030 7709050 7709070 770909L1 77091111 77091 3D 770915D 770917D 770919H 770921H 770923D 770925D 7709270 77092911 7709318 7709330 770935D 770937D 770939D 770941H 770943D 7709450 770947D /70949D 770951D 770953D 770955D 770957H 7709S9II 770961H 7709631' 7709651,1 770967H /70V69H 770971D 770973E' 770975D .'709/7D i 70979D 770981D 770V83D 770985D 770987D 7709898 770991D 7709930 / 70 9 9 5 El 7709V7D 7 709998 E 95 E. 97 t 9V E 101 E103 El 05 E 107 1109 El 11 El 13 e;iis e: 117 M39 E. 121 E 123 1.1 25 E 127 E 129 E.l 31 El 33 E.l 35 E 137 E.l 39 E141 E143 E145 E 147 E 1 49 E.l 51 E.l 53 fc 155 fc 157 E 159 E 161 E.l 63 E 3.65 E 167 El 69 El 71 El 73 E.l 75 E 177 E179 fc 181 fc 3 83 E 185 E.l 87 e i By E 191 E 1 93 ei yc E 1 97 fc i 99 CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) (0) CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) (0) CO) CO) <0> (0) CO) (0> CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) 3 7V 3 7V 179 181 181 181 183. 181 183 183 183 183 ie3 ie5 185 3 85 185 185 187 187 J 87 187 187 189 3 8V 189 189 ] 89 391 191 1 91 191 191 1 93 3 93 193 193 1 V3 1 V5 3 95 195 195 1 V5 19/ 197 3 97 197 1 97 199 3 99 1 99 1 99 1 99 K2I 1 K/l 2 R3 /* K1 R2 LI 1, 2 Rll. 1 R1L 2 K2L 1 R2L2 R3 7 R.1 F;2 L3 12 Kll 1 R1L 2 R2L1 R21 2 R3 7 R1 R2 l3 12 RU 1 K1 L 7 K2L1 K?l 2 K3 7 R1 K2 1l 1 Kill K3L2 R2L1 R2L2 K3 7 kl R2 LI 12 Kll 1 Kll 2 fc' 1 1 fc?l 3 fc: ,5 UAL MAI tTF.) - 500 HJ7AI FfcKAIfL 000 ,\ssuy. -i VO/I NO ROSS REFERENCE LI S-T I N G ILJ SAY NO. 126 0 HALES CO MR 780002 780004 780006 7S0008 780010 ' 780012 780014 780016 780018 780020, 780022 780024 7B0026 780028 780030, 780032 780034 780036 780038 780040 780042 780044 780046 780048 780050 780052 780054 780056 780058 780060 780062 780064 780066 7B0068 780070, 780072 760074 780076 780078 780080. 780082 780084 780086 780088 7800?0_ 780092 780094 780096 780098 780100 EXFR An A4 A6 A8 A 10 A 12 A 14 A 16 A 18 A 20 B2 84 86 88 8 10 8 12 8 14 B 16 8 18 B 20 C2 C4 C6 C8 C 10 C 12 C 14 c 16 c 18 c 20 n2 D4 B6 II 8 D 10 D 12 n 14 n 1t b 18 8 20 Eo EA E6 E8 E 10 E 12 E 14 E 16 E 18 F 20 SGF (0) (0) CO) (0) (0) CO) CO) CO) CO) CO) CO) CO) CO) (0) CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) <0) CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) CO) (0) CO) CO) CO) CO) CO) (0) CO) . <0> CO) CAGE EARFc 2 2 n 4- n 4 4 4 A 4 6 6 6 6 6 8 8 8 .8 8 10 10 10 10 10 12 12 12 12 12 14 14 14 14 14 16 16 16 16 16 18 18 18 18 18 20 20 20 20 20 Z Rl * R2 LI L2 R1L 1 R1L2 R2L1 R2L2 R3_ z Rl R2 LI L2_ R1L1 R1L2 R2L1 R2L2 z Rl R2 LI L2_ R1L1 R1L2 R2L1 R2L2 z Rl R2 LI L2 R1L 1 R1L2 R2L1 R2L2 R3_ Z Rl R2 LI L2 R1L 1 R1L2 R2L1 R2L2 R3 TOTAI MALES SO FEMALES COMR EXFR SGF CAGE E ARK1 780001 780003 780005 780007 780009 7800U 780013 780015 780017 780019, 780021 780023 780025 ' 780027 780029 780031 780033 780035 780037 780039 780041 780043 780045 780047 780049 780051 780053 780055 780057 780059 78006*1* 780063 780065 780067 780062, 780071 780073 780075 780077 780079 780081 780083 780085 780087 78008.9. 780091 78)093 780095 780097 780099 A1 A3 A5 A7 A9 A 11 A 13 A 15 A 17 A 19 h1 B3 B5 B7 B9 B 11 B 13 B 15 B 17 B 19 C1 c3 c5 c7 c9 c 11 c 13 c 15 c 17 c 19 r> 1 B3 B5 I> 7 B9 II 11 D 13 B 15 B 17 P 19 E1 E3 E5 E7 E9 E 11 E 13 E 15 E 17 E 19 CO) 1 2 (0) 1 Rl CO) ' 1 R2 CO) 1 LI CO) 1 ` L2. CO) 3 R1L1 CO) 3 R1L2 (0) 3 R2L1 CO) 3 R2L2 CO) 3 CO) 5 RJyL A. CO) 5 Rl CO) 5 R2 CO) 5 LI (0) 5 L? CO) 7 R1L1 CO) 7 R1L2 CO) 7 R2L1 (0) 7 R2L2 (0) 7 R3^ CO) 9 Z CO) 9 R1 (0) 9 R2 (0) 9 LI (0) 9 L2_ CO) 11 R1L1 CO) 11 R1L2 CO) 11 R2L1 CO) 11 R2L2 CO) 11 RZL. CO) 13 T' CO) 13 Rl CO) 13 R2 (0) 13 LI CO) 13 L2_ CO) 15 R1L 1 CO) 15 R1L2 CO) 15 R2L1 CO) 15 R2L2 CO) 15 R3 CO) 17 I CO) 17 R1 CO) 17 R2 CO) 17 LI CO) 17 L2_ CO) 19 R1L 1 (0) 19 R1L2 CO) ) 9 R2L 1 CO) 19 R2L2 CO) 19 R3 TOTAL FEMALES = R&S 040801