Groups of 60 Wistar rats of each sex were fed diets containing 3, 6 or 12% of the margarine emulsifier TOSOM (thermally oxidized soybean oil interacted with mono- and diglycerides of fatty acids) for 2.5 yr. In addition, three groups of 60 rats of each sex were fed two products of the release agent TOS (thermally oxidized soybean oil) in dietary levels of 1.2% TOS(G) (TOS from Grindsted Product A/S, Denmark) and 0.3 and 1.2% TOS(N) (TOS from Nexus Aps, Denmark), respectively for 2.5 yr. 120 rats of each sex fed a diet containing mono- and diglycerides served as controls. The diets given to all groups were isocaloric. Clinical appearance, food consumption, body weight and weight gain, survival, haematology, and clinical chemistry parameters were examined. Gross and histopathological examinations, including neoplastic and non-neoplastic lesions, were performed on all groups. Time to occurrence of tumours was recorded. No substance-related effect, including carcinogenicity, was found.

Groups of 60, 40, 40 and 60 F0 Wistar rats of each sex were fed a semi-synthetic diet containing butylated hydroxytoluene (BHT) in concentrations to provide intakes of 0, 25, 100 or 500 mg/kg body weight/day, respectively. The F0 rats were mated and groups of 100, 80, 80 or 100 F1 rats of each sex were formed from 40, 29, 30 and 44 litters, respectively. After weaning, the highest dose (500 mg BHT/kg/day) was lowered to 250 mg/kg/day for the F1 rats. The numbers of litters of ten or more pups at birth decreased with increasing BHT dose. At weaning, treated F1 rats had lower body weights than the controls, the extent of the reduction being dose related; the effect, which persisted throughout the study, was most pronounced in the males. The survival of BHT-treated F1 rats of both sexes was significantly better than that of the controls. No significant changes attributable to BHT treatment were found in the haematological parameters. F1 females on the highest dose showed an increase in serum cholesterol and phospholipids, and serum triglycerides were reduced in this group in both sexes. Dose-related increases in the numbers of hepatocellular adenomas and carcinomas were statistically significant (at P less than 0.05 or lower) in male F1 rats when all groups together were tested for heterogeneity or analysis for trend. The increase in hepatocellular adenomas and carcinomas in treated female F1 rats was only statistically significant for adenomas (at P less than 0.05) in the analysis for trend. All hepatocellular tumours were detected when the F1 rats were more than 2 yr old. Tumours were found in many other organs of some of the treated rats, but their incidence was not significantly different from that in controls. The role of BHT in the development of hepatocellular tumours requires further elucidation.

Butylated hydroxyanisole (BHA) was given to pregnant SPF pigs (Danish Landrace) in doses of 0, 50, 200 and 400 mg/kg body weight/day from mating to day 110 of the gestation period. The BHA was mixed in the diet (pelleted). Caesarean section was performed on gestation day 110. BHA affected neither the reproduction data nor the incidence of defects in the foetuses. Significantly lower weight gain was observed in the group of dams on the highest dose. Absolute and relative organ weights for the liver and thyroid gland showed a dose-related increase. Proliferative and parakeratotic proliferative changes of the stratified epithelium of the stomach were found in both control and treated pigs. In addition, proliferative and parakeratotic changes of the oesophageal epithelium were observed in a few pigs in the two groups on the highest doses. Papillomas were not found, and no changes of the glandular part of the stomach were observed.

Groups of 40, 29, 39 and 44 F0 rats of each sex were fed a semi-synthetic diet containing butylated hydroxytoluene (BHT) in concentrations to provide intakes of 0, 25, 100 or 500 mg/kg body weight/day, respectively. The F0 rats were mated, and groups of 100, 80, 80 and 100 F1 rats of each sex were formed. After weaning, the highest dose of BHT was lowered to 250 mg/kg/day for the F1 rats. At weaning the BHT-treated F1 rats, especially the males, had lower body weights than the controls and the effect was dose related. The survival of the BHT-treated rats of both sexes was higher than that of the controls. Dose-related increases in the numbers of hepatocellular adenomas and carcinomas were statistically significant in male F1 rats when all groups together were tested for heterogeneity or analysis for trend. The increases in hepatocellular adenomas and carcinomas in treated female F1 rats were only statistically significant for adenomas in the analysis for trend. All hepatocellular tumours were detected when the F1 rats were more than 2 years old.

In order to detect possible formation of carcinogenic N-nitroso compounds from nitrite and nitrosatable compounds in meat, studies were carried out with 70 male and 140 female F0 rats, divided into six groups, and 60, 100, 70, 60, 60 and 66 of their male and female offspring. One control group received casein and other groups chopped pork as the sole protein source (45%, mass/mass) on a fresh basis, either salted (sodium chloride) or not. For test groups, nitrite was also added to the meat before autoclaving and storing the diet and represented mass fractions of 200, 1 000 and 4 000 mg/kg, as sodium nitrite. The results do not demonstrate any effect on reproduction and no significant carcinogenic effect was revealed. However, an observed tendency toward an increased number of tumour-bearing rats in the highest dose group, plus the possible formation of carcinogenic N-nitroso compounds in nitrite-treated meat products, led to a recommendation to reduce the use of nitrite. Results from a concomitant study demonstrate that it is possible to produce many cured-meat products with the addition of only 50 mg/kg nitrite.

The oral LD50 for malachite green oxalate was found to be 275 mg/kg in rats while the approximate lethal dose for NMRI mice was 50 mg/kg. No systemic effects were seen after dermal application of 2,000 mg/kg. Repeated administration in the diet for 28 days to rats produced only minor changes in serum urea and aspartate aminotransferase levels. The rats at the highest dose level showed decreased weight gain and appeared clinically to have elevated motor activity. No sex differences were observed in either acute or prolonged experiments. In accord with human experience malachite green was irritating to mucous membranes, but no effects were seen on intact skin nor was it shown to be sensitizing. It was found to be a mutagen in the Salmonella/microsome test after metabolic activation but without clastogenic activity when tested at maximally tolerated levels in mice in the micronucleus test.

Acta Pharmacologica et ToxicologicaVolume 53, Issue 5 p. 433-434 Hepatocellular Neoplasms in Rats Induced by Butylated Hydroxytoluene (BHT) Preben Olsen, Preben Olsen From the Institute of Toxicology, National Food Institute, Mørkhøj Bygade 19, DK-2860 Søborg, DenmarkSearch for more papers by this authorNils Bille, Nils Bille From the Institute of Toxicology, National Food Institute, Mørkhøj Bygade 19, DK-2860 Søborg, DenmarkSearch for more papers by this authorOtto Meyer, Otto Meyer From the Institute of Toxicology, National Food Institute, Mørkhøj Bygade 19, DK-2860 Søborg, DenmarkSearch for more papers by this author Preben Olsen, Preben Olsen From the Institute of Toxicology, National Food Institute, Mørkhøj Bygade 19, DK-2860 Søborg, DenmarkSearch for more papers by this authorNils Bille, Nils Bille From the Institute of Toxicology, National Food Institute, Mørkhøj Bygade 19, DK-2860 Søborg, DenmarkSearch for more papers by this authorOtto Meyer, Otto Meyer From the Institute of Toxicology, National Food Institute, Mørkhøj Bygade 19, DK-2860 Søborg, DenmarkSearch for more papers by this author First published: November 1983 https://doi.org/10.1111/j.1600-0773.1983.tb03447.xCitations: 20AboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinked InRedditWechat No abstract is available for this article.Citing Literature Volume53, Issue5November 1983Pages 433-434 RelatedInformation

Butylated hydroxyanisole (BHA) was given to pregnant SPF pigs (Danish Landrace) in doses of 0, 50, 200 and 400 mg/kg body wt per day for 110 days. BHA was mixed in the diet (pelleted). Hyperkeratotic and parakeratotic proliferative changes of the stratified epithelium of the stomach were found at various degree in both control and dosed pigs. Papillomas were not found and no changes of the glandular part of the stomach were observed.

An embryotoxicity study on butylated hydroxyanisole (BHA) was carried out in SPF pigs (Danish Landrace). BHA was incorporated in the diet and administered to pigs in doses of 0, 50, 200 and 400 mg/kg body wt/day from mating (artificial insemination) to day 110 of the gestation period, when the foetuses were removed. Significant lower weight gain was observed in the dams dosed 400 mg/kg body wt/day. Absolute and relative organ weight for the liver and thyroid gland showed a dose-related increase, BHA neither affected the reproduction data nor the incidence of defects in the foetuses.

Orange RN (monosodium salt of 1-phenylazo-2-naphthol-6-sulphonic acid) was fed to pigs at dietary levels of 160, 40, 10 and 0 (control)_mg/kg body weight/day for 15 weeks. In the 160-mg group hepatosis with liver enlargement, fibrosis and bile-duct proliferation was observed. Proliferation of the cells of the bile-ductule epithelium was found in all the test groups and the intensity of proliferation was dose-related. At the highest dose level macrocytic anemia, hemiglobinemia and increase in ASAT and LD serum levels were observed. Heinz-body formation was marked in the groups fed 160 and 40 mg/kg body weight. The study indicates the pigs are very sensitive to the effect of Orange RN on the liver. A no-effect-level has not been found in these experiments, but it is lower than 10 mg/kg body weight/day.

Acta Pharmacologica et ToxicologicaVolume 32, Issue 3-4 p. 314-316 Bile Duct Proliferation in Pigs Fed the Food Colour Orange RN Preben Olsen, Preben Olsen Institute of Toxicology, National Food Institute, DK-2860 Søborg, DenmarkSearch for more papers by this authorErnst Hansen, Ernst Hansen Institute of Toxicology, National Food Institute, DK-2860 Søborg, DenmarkSearch for more papers by this author Preben Olsen, Preben Olsen Institute of Toxicology, National Food Institute, DK-2860 Søborg, DenmarkSearch for more papers by this authorErnst Hansen, Ernst Hansen Institute of Toxicology, National Food Institute, DK-2860 Søborg, DenmarkSearch for more papers by this author First published: March 1973 https://doi.org/10.1111/j.1600-0773.1973.tb01476.xCitations: 3AboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onEmailFacebookTwitterLinkedInRedditWechat References Dacre, J. L.: Acute and chronic toxicity studies on Orange RN. Proc. Univ. Otago med. Sch. 1969, 47, 3. Web of Science®Google Scholar Gaunt, I. F., P. G. Brantom, I. S. Kiss, P. Grasso & S. D. Gangolli: Short-term toxicity of Orange RN in rats. Fd. Cosmet. Toxicol. 1971, 9, 619–630. 10.1016/0015-6264(71)90149-0 CASPubMedWeb of Science®Google Scholar Sisk, D. B., W. W. Carlton & T. M. Curtin: Experimental aflatoxicosis in young swine. Amer. J. Vet. Res. 1968, 29, 1591–1602. CASPubMedWeb of Science®Google Scholar Citing Literature Volume32, Issue3-4March 1973Pages 314-316 ReferencesRelatedInformation