NATIONAL TOXICOLOGY PROGRAM Technical Report Series No. 337 TOXICOLOGY AND CARCINOGENESIS STUDIES OF NITROFURAZONE (CAS NO. 59-87-0) IN F344/N RATS AND B6C3F1 MICE (FEED STUDIES) U.S. DEPARTMENT OF HEALTH AND HUMAN SERVICES Public Health Service National Institutes of Health NATIONAL TOXICOLOGY PROGRAM The National Toxicology Program (NTP), established in 1978,develops and evaluates scientific information about potentially toxic and hazardous chemicals. This knowledge can be used for protecting the health of the American people and for the primary prevention of disease. By bringing to- gether the relevant programs, staff, and resources from the U.S.Public Health Service, DHHS, the National Toxicology Program has centralized and strengthened activities relating to toxicology research, testing and test developmenthalidation efforts, and the dissemination of toxicological in- formation to the public and scientific communities and to the research and regulatory agencies. The NTP is made up of four charter DHHS agencies: the National Cancer Institute (NCI), National Institutes of Health; the National Institute of En- vironmental Health Sciences (NIEHS), National Institutes of Health; the National Center for Toxicological Research (NCTR), Food and Drug Ad- ministration; and the National Institute for Occupational Safety and Health (NIOSH), Centers for Disease Control. In July 1981,the Carcino- genesis Bioassay Testing Program, NCI, was transferred to the NIEHS. Nitrofurazone, NTP TR 337 NTP TECHNICAL REPORT ON THE TOXICOLOGY AND CARCINOGENESIS STUDIES OF NITROFURAZONE (CAS NO. 59-87-0) IN F344/N RATS AND B6C3F1 MICE (FEED STUDIES) Frank Kari, Ph.D., Chemical Manager NATIONAL TOXICOLOGY PROGRAM P.O. Box 12233 Research Triangle Park, NC 27709 June 1988 NTP TR 337 NIH Publication No. 88-2593 U.S. DEPARTMENT OF HEALTH AND HUMAN SERVICES Public Health Service National Institutes of Health NOTETOTHEREADER This study was performed under the direction of the National Institute of Environmental Health Sci- ences as a function of the National Toxicology Program. The studies described in this Technical Re- port have been conducted in compliance with NTP chemical health and safety requirements and must meet or exceed all applicable Federal, state, and local health and safety regulations. Animal care and use were in accordance with the U.S.Public Health Service Policy on Humane Care and Use of Ani- mals. All NTP toxicology and carcinogenesis studies are subjected to a data audit before being pre- sented for public peer review. Although every effort is made to prepare the Technical Reports as accurately as possible, mistakes may occur. Readers are requested to identify any mistakes so that corrective action may be taken. Further, anyone who is aware of related ongoing or published studies not mentioned in this report is encouraged to make this information known to the NTP. Comments and questions about the National Toxicology Program Technical Reports on Toxicology and Carcinogenesis Studies should be directed to Dr. J.E. Huff,National Toxicology Program, P.O.Box 12233, Research Triangle Park, NC 27709 (919-541-3780). These NTP Technical Reports are available for sale from the National Technical Information Service, U.S.Department of Commerce, 5285 Port Royal Road, Springfield, VA 22161 (703-487-4650). Single copies of this Technical Report are available without charge (and while supplies last) from the NTP Public Information Office, National Toxicology Program, P.O. Box 12233, Research Triangle Park, NC 27709. Nitrofurazone, NTP TR 337 2 THIRTEEN-WEEK STUDIES .. ... .... .. ... .. .. ..46 TWO-YEAR STUDIES , .. .. .e.* .. s. s 8 . 41 BODY WEIQHTS, FEED CONSUMPTION, AND CLINICAL SIGNS ,,...... , , , , ....,.,41 SURVIVAL s s e s e e e e e s s s s s e s 8 8 e 9 s s e s s e 8 s 0 8 s e s e M) PATHOLOQY AND STATISTICAL ANALYSES OF RESULTS ,.,,.. ...... .. .. sa 3 Nitrofurazone, NTP TR 337 CONTENTS (Continued) PAGE IV. DISCUSSION AND CONCLUSIONS .................................................... 57 V. REFERENCES ..................................................................... 6s APPENDIXES APPENDIX A SUMMARY OF LESIONS IN MALE RATS IN THE TWO-YEAR FEED STUDY OF NITROFURAZONE .............................................. 69 APPENDIX B SUMMARY OF LESIONS IN FEMALE RATS IN THE TWO-YEAR FEED STUDY OF NITROFURAZONE .............................................. 91 APPENDIX C SUMMARY OF LESIONS IN MALE MICE IN THE TWO-YEAR FEED STUDY OF NITROFURAZONE ............................................. 111 APPENDIX D SUMMARY OF LESIONS IN FEMALE MICE IN THE TWO-YEAR FEED STUDY OF FWI"'I'OFURAZONE ............................................. 135 APPENDIX E GENETIC TOXICOLOGY OF NITROFURAZONE .............................. 159 APPENDIX F SENTINEL ANIMAL PROGRAM ............................................ 165 APPENDIX G FEED AND COMPOUND CONSUMPTION BY RATS AND MICE IN THE TWO-YEAR FEED STUDIES OF NITROFURAZONE ........................... 169 APPENDIX H INGREDIENTS. NUTRIENT COMPOSITION. AND CONTAMINANT LEVELS IN NIH 07 RAT AND MOUSE RATION ......................................... 175 APPENDIX I AUDIT SUMMARY ....................................................... 181 Nitrofurazone.NTP TR 337 4 NITROFURAZONE CAS NO. 59-87-0 C6H6N404 Molecular weight 198.1 Synonymsor Trade Names: 5-Nitro-2-furaldehyde semicarbazone; 2-[(5-Nitro-2-furanyl)methylenelhydrazinecarboxamide; Aldomycin; Amifur; Chemfuran; Coxistat; Furacin; Furacinetten; Furaplast; Furazol W; Furesol; Furracoccid; Mammex; Nefco; Nifuzon; Nitrofural; Vabrocid ABSTRACT Nitrofurazone is a synthetic furan derivative, active against a broad spectrum of bacteria, which has been widely used in veterinary and human medicine. Toxicology and carcinogenesis studies were conducted by feeding diets containing nitrofurazone (99% pure) to groups of F344/N rats and B6C3F1 mice of each sex for 14 days, 13weeks, or 2 years. Fourteen-Day and Thirteen-Week Studies: Groups of five males and five females of each species were fed diets containing 0, 630, 1,250, 2,500, 5,000, or 10,000 ppm for 14 consecutive days. Early deaths occurred in all groups of rats receiving 5,000 or 10,000ppm nitrofurazone. The surviving rats in the lower two dose groups gained weight, but weight gain was decreased as the dose of nitrofurazone was increased. Feed consumption by rats of each sex was decreased at all doses above 630 ppm. In all dosed groups, clinical signs of toxicity included rough hair coats and lethargy. At doses of 2,500 ppm and above, rats of each sex exhibited intermittent episodes of seizures and lethargy. All mice that received 2,500, 5,000, or 10,000 ppm nitrofurazone and 315 males that received 1,250 ppm died before the end of the 1Cday studies; the surviving dosed mice (except females at 630 ppm) lost weight. A dose-related decrease in feed consumption was observed at all doses above 630 ppm. Clinical signs included rough hair coats and convulsive seizures. In the 13-week studies, groups of 10 rats of each sex were given diets containing 0, 150, 310, 620, 1,250,or 2,500 ppm nitrofurazone, No deaths were observed and all animals gained weight, but the magnitude of weight gain was dose dependent with decrements in final mean body weight for the highest dose group reaching 55% in males and 36% in females. Other evidence of chemically related toxicity included convulsive seizures, osteoporosis, degenerative arthropathy, and gonadal hypo- plasia in both sexes at the two highest doses. Groups of 10mice of each sex were given diets containing 0,70, 150,310,620, or 1,250 ppm nitrofura- zone for 13weeks. Early deaths were observed in the two highest dose groups of each sex. The final mean body weights of male and female mice in the 1,250-ppm groups were about 20% lower than those of the controls; weight gains of the other dosed mice were comparable to those of the controls. Stimulus-induced convulsive seizures were observed for all mice in the two highest dose groups. Tes- ticular hypoplasia was observed in the two highest dose groups of male mice. Body Weight and Survival in the Two-year Studies: Dietary concentrations for the 2-year studies were 0,310, or 620 ppm for rats and 0,150, or 310 ppm for mice (60 animals per dose group). Mean 5 Nitrofurazone, NTP TR 337 body weights of high dose male rats were lower than those of the controls after week 39; mean body weights of low dose male rats and of the controls were comparable throughout the study. Final mean body weights of low and high dose female rats were 9% and 21% lower than those of the controls. Dosed rats consumed less feed than did the controls. The average amount of nitrofurazone consumed per day was approximately 11-12 or 24-26 mg/kg by low or high dose male and female rats. The sur- vival of the high dose group of male rats was lower than that of the controls after week 92 (final sur- vival--male: control, 33/50; low dose, 30/50; high dose, 20150; female: 28/50; 37/50; 31/50). Mean body weights of dosed mice were similar to or somewhat greater than those of the controls throughout most of the studies. The average daily feed consumption by dosed mice was similar to that of controls. The average amount of nitrofurazone consumed per day was approximately 14-16 or 29-33 mg/kg for low or high dose male and female mice. The survival of the high dose group of male mice was lower than that of the controls after week 88 (final survival--male: 39/50; 31/50; 27/50; fe- male: 39/50; 40/50; 35/50). In mice of each sex, nitrofurazone administration induced stimulus-sensitive convulsive seizures be- ginning at week 4 or 5 for high dose mice and week 24 for low dose female mice. These seizures were observed primarily in the first year of the study. Nonneoplastic and Neoplastic Effects in the Two-year Studies: Degenerative changes involving the vertebral and femoro-tibial (knee) joints were observed at increased incidences in dosed rats. The de- generative changes primarily affected the articular cartilage and were similar to those seen in the 13- week studies. Degeneration of the sternal synchondroses was increased in high dose female rats.