FLUSILAZOLE EXPLANATION Flusilazole is a fungicide. It was reviewed for the first time at the present meeting. EVALUATION FOR ACCEPTABLE DAILY INTAKE BIOLOGICAL DATA Biochemical aspects Absorption, distribution and excretion 14C-Flusilazole (uniformly phenyl labelled) was administered orally to two male and two female Charles River CD rats/group by one of three regimens: 1) a single dose of about 8 mg/kg bw with no preconditioning: 2) a single dose of about 8 mg/kg bw following preconditioning for 21 days on a diet containing 100 ppm flusilazole; or 3) a single dose of about 200 mg/kg bw with no preconditioning. By 96 (regimen 1) and 168 hours (regimens 2 and 3), 90% of the administered radioactivity was recovered in the urine and feces in both sexes. Preconditioning did not affect excretion. Excretion t1/2 was 31 hours in males and 37 hours in females. There was a sex difference in the amount of material excreted in feces compared to urine. In males total fecal recovery accounted for 87% of the dose with 8% in urine, but in females the corresponding figures were 59% and 23%. No significant amount of radioactivity was expired as CO2 or volatile metabolites. Tissue residues accounted for less than 25% of the dose with highest amounts in carcass, GI tract and liver (means less than 1%). An additional 1 male and 1 female rat were given a single dose of 14C-flusilazole (triazole-3-labelled) of about 8 mg/kg bw without preconditioning. As with the phenyl-labelled material, 88% of the administered radioactivity was recovered in urine and feces within 96 hours. However, the pattern of excretion was different with the predominant route being the urine. In males, urinary radioactivity accounted for 78% of the dose, with 11% in feces; in females, the respective figures were 59% and 26%. Total tissue residues were about 3% of the dose with the highest levels in carcass (approx. 2% of dose), skin, GI tract and liver (all less than .5% of dose (Anderson et al. 1986). In another study, 14C-flusilazole (triazole-3-labelled) was given orally to groups of 5 male and 5 female Charles River Crl:CD(SD)BR rats by one of the following regimens: 1) a single dose of about 8 mg/kg bw without preconditioning; 2) a single dose of about 8 mg/kg bw following a 14-day treatment period with doses about 8 mg non-labelled material/kg bw/day by gavage; 3) a single dose of about 224 mg/kg bw without preconditioning. Total recovery of radioactivity at 96 hours (regimen 1) or 120 hours (regimens 2 and 3) was 92.6 to 99.2%. Sex and regimen differences were not observed in this study. Urinary excretion accounted for about 72% and fecal excretion for 17%. About 90% of the material excreted was recovered in the first 48 hours with all regimens. Tissue levels were low and excluding carcass were less than .2% of the dose. Carcass residues accounted for less than 3% of the dose (Cheng, 1986). Biotransformation 14C-Flusilazole was extensively metabolized when administered orally to Charles River CD rats. Recovered parent compound accounted for 2-11% of the dose in all groups of animals and was round almost exclusively in feces; urinary levels were less than 1% of the dose. The chief fecal metabolites identified following dosing with phenyl labelled flusilazole were: [Bis(4-fluorophenyl)methyl] silanol (Metabolite 1) (about 30% of dose in males, 19% in females); [Bis(fluorophenyl)methylsily] methanol (Metabolite 2a) (about 9% of dose in both sexes); fatty acid conjugates of the latter (19% of dose in males, 10% in females); and disiloxane (about 11% of dose in males, 7% in females). The same metabolites except for the fatty acid conjugates were found in urine but in smaller amounts; none exceeded 1% of the dose in males. In females, urine contained 7.5% of the dose as metabolite 1 and 2.2% of the dose as metabolite 2a, and 1.9% of the dose as siloxane. Following dosing with triazole labelled flusilazole, the chief metabolite identified was 1H,1,2,4-triazole (metabolite 3) which was found predominantly in urine (63.8% of dose in males, 51.6% in females) with smaller amounts in feces (4.0% of dose in males, 17.2% of dose in females). The proposed metabolic pathway is shown in Figure 1 (Anderson, 1986). Metabolite 1: [Bis(4-fluorophenyl)methyl]silano Metabolite 2a [Bis(4-fluorophenyl)methyl silyl]methanol Metabolite 2 1-0-[Bis(4-fluorophenyl)methyl silyl]methyl]-ß-D- glucopyranuronic acid Metabolite 3 1H-1,2,4-Triazole Metabolite 4 N-((4-fluorophenyl)methyl) glycine.Toxicological studies Acute toxicity The acute toxicity of flusilazole to several animal species is given in Table 1. Signs of toxicity following oral administration included weight loss, weakness, lethargy and, at higher doses, laboured breathing, convulsions and prostration. Inhalation effects were chiefly laboured breathing and lung sounds. Dermal administration resulted only in some local erythema. TABLE 1. RESULTS OF ACUTE TOXICITY ASSAYS WITH FLUSILAZOLE SPECIES SEX ROUTE LD50a/ ALDa/ REFERENCES LC50b ALCb Technical flusilazole Rat M oral 1500 Wylie et al. 1983 Rabbit M&F oral 350 Redgate et al. 1985 Mouse M oral 680 Wylie et al. 1985 F oral 1000 Wylie et al. 1985 Rat M oral 1110 Wylie et al. 1984a F oral 674 Wylie et al. 1984a Rabbit M&F dermal >2000 Gargus et al. 1983 Rat M inhal. 2.7b,c Poindexter et al. 1984 F inhal. 3.7b,c Poindexter et al. 1984 Rat M&F inhal. >5.0b Turner et al. 1985 Formulation - 40% flusilazole Rat M oral 1865 Wylie et al. 1984b F oral 1272 Wylie et al. 1984b a mg/kg bw b mg/l c test material was 99% pure. Short-term studies Oral studies Rat Groups of six male Crl:CD rats were given 0 or 300 mg flusilazole (95.5%) in corn oil/kg bw/day by oral gavage for 2 weeks (5 days per week). After the tenth dose 3 rats/group were killed and examined for histopathology. The remaining rats were killed after a 14-day recovery period. One treated rat died after the fifth dose (found Day 7). The treated rats died lost weight during the first week of treatment and showed alopecia, diarrhea, stained face and perineal area, salivation and hypersensitivity during the treatment period only. Liver vacuolation was seen in all treated animals. In urinary bladder transitional epithelium hyperplasia and vacuolation were seen in all sacrificed treated animals with an increase in mitotic figures seen in two of three males killed immediately following the treatment period. Two treated males had atypical giant cells in the lumen of the seminiferous tubules and necrosis and cellular degeneration of the germinal epithelium in the testes. Hyperplasia and vacuolation of the transitional epithelium in the kidney pelvis was seen in two treated males. Lesions tended to be more severe in animals killed immediately after the treatment period than in those from the recovery phase (Wylie et al. 1984c). A 90-day feeding study was carried out in Charles River (CD) rats with dietary levels of flusilazole (96.7%) of 0, 25, 125 and 375 ppm fed to groups of 16 rats/sex and 750 ppm fed to 10 rats/sex. Groups of 10/sex/dose comprised the standard 90-day study while the extra 6/sex/dose continued on diet for a one generation one litter reproduction study and then received control diet for a 4-month recovery phase. Body weight gains were reduced only in the females given 750 ppm. There were no effects on food consumption, clinical signs of toxicity, hematological parameters or urinalyses. Serum cholesterol levels were increased in both males and females given 750 ppm. Absolute and relative liver weights were increased in both sexes at 750 ppm. Mild urothelial hyperplasia was observed in urinary bladders of 5 males and 8 females at 750 ppm and 1 male and 4 females at 375 ppm. After the 4-month recovery phase following dosing with 0-375 ppm flusilazole, no evidence of bladder hyperplasia was observed. At 750 ppm 5 males had liver lesions, chiefly hepatocyte hypertrophy in 5 males and fatty change/hepatocytolysis in 4 males. The NOAEL was 125 ppm (equal to 10 mg/kg bw/day) (Pastoor et al. 1983). Mouse Groups of 20 male and 20 female Crl:CD-1 mice were given diets containing 0, 25, 75, 225, 500 or 1000 ppm of flusilazole (96.7% pure). After four weeks on these diets 10 mice/sex/group were sacrificed. The remaining 10 mice/sex/group were sacrificed after 90 days of treatment. There were no effects noted on body weight, food consumption or clinical signs of toxicity in animals sacrificed at either interval. At 90 days high-dose males and females had slightly reduced Hb, Hct and RBC counts which were statistically significant for at least one of these parameters in each sex. No other hematological changes were noted. At both 4 weeks and 90 days absolute and relative liver weights were increased in male mice at 225 ppm and higher and in female mice at 75 ppm and higher. The absolute and relative kidney weights in males at 1000 ppm were decreased. At 90 days hepatocellular hypertrophy was observed in 4/10 females at 1000 ppm and 8-10/10 males at 225, 500, and 1000 ppm. Hepatocellular vacuolar cytoplasmic changes were observed in both sexes (incidences 0, 0, 0, 2, 7, 10 in males and 0, 0, 1, 3, 9, and 8 in females, at 0, 25, 75, 225, 500, and 1000 ppm, respectively). Urothelial cell hyperplasia in urinary bladder was observed in 1/10 males and 7/10 males at 500 and 1000 ppm, respectively, and 2/10, 3/10, and 6/10 females at 225, 500 and 1000 ppm, respectively. No treatment-related kidney lesions were observed. Tissues from animals killed at 4 weeks were not examined. The NOAEL in this study was 25 ppm (equal to 4.5 mg/kg bw/day) (Pastoor et al. 1984). Dog Four beagle dogs/sex/group were given diet containing 0, 25, 125, or 750/500 ppm flusilazole (93% pure) for 3 months. At the highest dose level the dogs were given diet containing 750 ppm flusilazole for 3 weeks, control diet for one week and then a dose level of 500 ppm for the remainder of the study. The dose level was reduced because of body weight loss and reduced food intake at 750 ppm. At 500 ppm the males gained a mean of 0.2 kg while the females lost a mean of 0.5 kg body weight. The other two groups were comparable to controls (gains of 1.6-1.9 kg in both sexes). Food consumption remained lower than controls at the 750/500 ppm dose level. Clinical signs related to treatment were observed in one male and two females at 500 ppm (weakness or tremors). One female at 25 ppm was sacrificed in extremis week 5. No other animal died. In males at 750/500 ppm WBC count and monocyte counts were slightly but statistically significantly increased and cholesterol levels, total protein and albumen levels were lower than controls. In both sexes at 750/500 ppm and males at 125 ppm alanine amino transerase levels were higher than controls. Absolute and relative liver weights were increased in both sexes at 750/500 ppm and were considered related to treatment. Other organ weight differences were probably related to the decreased body weights at 750/500 ppm. Mild mucosal hyperplasia in the urinary bladder was observed in all males and females at 500 ppm and in 2 males at 125 ppm. In the stomach, hyperlasia of lymphoid follicles in the pyloric granular mucosa was observed in 0/4, 3/4, 4/3, and 4/4 males and 0/4, 0/4, 3/4, and 4/4 females at 0, 25, 125, and 750/500 ppm, respectively. On the basis of this effect in stomach a NOAEL was not demonstrated in this study (Richard et al. 1983). Groups of 5 beagle dogs/sex were given diets containing 0, 5, 20, or 75 ppm flusilazole (95.8% pure) for one year. Although all treated groups of females had lower weight gains than controls, this did not appear to be treatment-related. No body weight effects were seen in males. No treatment-related effects were noted in food consumption or clinical signs of toxicity. WBC counts were higher in both sexes at 75 ppm than in the controls. Alkaline phosphates levels were higher than controls and cholesterol levels lower in both sexes at 75 ppm but the difference was statistically significant only in males. The 75 ppm males also had lower total protein and albumen levels in serum. Relative liver weights were increased in both sexes at 75 ppm and relative kidney weight in females at 75 ppm. There was centrilobular hepatocyte enlargement at 20 and 75 ppm, in both sexes with distinct vacuolation in 3 high-dose males. There were also a higher degree of centrilobular inflammation in all of the high-dose dogs both with regard to relative numbers of inflammatory cells and to increased number of hepatic veins involved. All dogs in all groups including controls had lymphoid hyperplasia in the gastric mucosa but the degree increased with dose level and was moderate only in high-dose males and females. There were no treatment-related kidney or urinary bladder effects. The NOAEL in this study was 5 ppm (equal to 0.14 mg/kg bw/day) (O'Neal et al. 1985). Dermal studies Rabbit A study in New Zealand White rabbits in which groups of 5-10 rabbits/sex were treated dermally with 0, 25, 250 or 2000 mg flusilazole/kg bw/day (95.8% pure material) was terminated after 6 days exposure because of serve erythema with necrosis in animals at 250 and 2000 mg/kg bw/day. The erythema was first observed Day 4 of test. A range-finding study with 175 or 250 mg flusilazole (94.9%) kg bw/day produced only mild erythema at both levels. A 21-day study was performed in groups of 5 New Zealand White rabbits/sex/dose at dose levels of 0, 1, 5, 25, and 200 mg flusilazole (94.9% pure)/kg bw/day. For application the test material was moistened with distilled water. The exposure sites were occluded for 6 hours and then washed free of remaining test material. There were no apparent treatment-related effects on body weights: hematology; clinical chemistry; or liver, kidney and spleen weights. At 200 mg/kg bw/day, four males and 2 females had persistent mild erythema beginning Days 6 to 12. At termination slight to mild diffuse hyperplasia/thickening of epidermis was observed in the skin of one male and two females at 25 mg/kg bw/day and three males and two females at 200 mg/kg bw/day (Sarver et al. 1986). Long-term/carcinogenicity studies Rats Groups of 70 male and 70 female Crl:CD(SD)BR rats were given diets containing 0, 10, 50 or 250 ppm flusilazole (95.6% pure) for two years. Additionally, groups of 10/sex were given the same dose levels for 6 months and 1 year. After about 100 days on test, 20 rats/sex/group were mated for the reproduction phase of the study. These rats were returned to the long-term study after weaning of the second litter. Body weights, food consumption and clinical condition were monitored throughout the study. Hematology and clinical chemistry parameters were examined at 3, 6, 9, 12 and 24 months on fasted animals. The animals sacrificed at 6 months were examined for bladder lesions only. About 40 tissues/organs were examined for control and high-dose animals and liver, bladder, kidney, lung and gross lesions for the low and intermediate dose groups sacrificed after one year. For animals that died on test or were sacrificed at two years all tissues from all animals in all groups were examined. Survival to two years was 39-57% in the various groups with no treatment-relationship. Survival did not drop below 50% in any group until Week 98. The high-dose males had slightly lower body weights from 6 months to the end of the study but there was no similar effect in females. There were no treatment-related effects on food consumption, clinical signs of toxicity, hematology, clinical chemistry or urinalysis. In females, relative liver weights were increased at both one and two years at 250 ppm and at one year only at 50 ppm. No effect was observed in males. No treatment-related effects on the urinary bladder were observed at 6 months. After one year there was an increased incidence of hepatocellular hypertrophy in females (0/10, 0/10, 1/10, and 7/10 at 0, 10, 50, and 250 ppm, respectively) and at 250 ppm polyploidy (increased number of multinuclear hepatocytes) was observed in two females with none in any other group. There were no liver changes in males at one year. There was an increased incidence of hydronephrosis in males (0/10, 0/10, 3/10, and 4/10 at 0, 10, 50 and 250 ppm). Urothelial hyperplasia in renal pelvis was observed in one male at 50 pm and one at 250 ppm, but not in any other group. Treatment-related lesions were not observed in urinary bladders of males. Females did not show kidney or bladder lesions after one year. Among rats dying on test or sacrificed after two years there were increased incidences in livers of females of acidophilic foci of cellular alterations (3, 3, 4, and 13 at 0, 10, 50, and 250 ppm, respectively) and diffuse fatty change (9, 9, 12, and 23, respectively, in 63-68 /group). There were no treatment-related liver lesions in males. In females, there was an increased incidence of pyelonephritis in kidney (3, 3, 8, and 10, respectively), but no treatment-related lesions were observed in males. The only tumours which were increased in incidence in treated groups were squamous cell carcinomas of the oral and nasal cavities in males (0/66, 1/63, 0/67, and 3/64, respectively). Historical control data from six two-year feeding studies showed five studies which no squamous cell carcinomas of oral or nasal origin while the remaining study had an incidence of 2/60. These data suggest that these tumours may occur randomly as clusters and that the incidence in this study was not treatment-related. There was, therefore, no evidence of oncogenicity. It is noted, however, that the effects noted were relatively mild and higher doses might have been tolerated. The NOAEL for non-neoplastic effects was 10 ppm (equal to 0.45 mg/kg bw/day) (Pastoor et al. 1986). Mouse Groups of 80 Crl:CD-1(ICR)BR mice/sex were given a diet containing 0, 5, 25, or 200 ppm flusilazole (93%-95.6% purity) for 18 months. Additional groups of 10/sex/dose were given the same diets for 6 months. Survival to 18 months was 76-86% in males and 57-80% in females with the highest survival at the high-dose level. There were no treatment-related effects on body weight, food consumption, hematology or serum protein. There was a higher incidence of skin sores in high-dose males and females but these were mainly ear lesions which were noted at the beginning of the study and appeared to be related to the animal identification process. High-dose males also had an increased incidence of ruffled fur. Absolute and relative liver weights were increased in males at 200 ppm at both 6 and 18 months and in 200 ppm females at 6 months. Absolute kidney weight only was decreased in females at 200 ppm at 18 months but the biological significance of this observation was not clear. There were no treatment-related lesions in the kidney. In the liver there was an increased incidence of hepatocellular fatty change in both males and females at 200 ppm at 18 months (40/80 and 24/80, respectively, compared to 4/80 and 2/79 in male and female controls. One 200 ppm male at 6 months also had this lesion, although 8/80 males at the 25 ppm levels also had hepatoceullar fatty changes. This incidence was not statistically significantly different from controls. There was no treatment- related increase in incidence of any tumour type in this study. The NOAEL in this study was 25 ppm (equal to 3.4 mg/kg bw/day) (Brock et al. 1985). Reproduction study Rat Groups of 6 Crl:CD(SD)BR rats/sex/dose level from the 90 day feeding study were mated within the same dose level for a period of 15 days. Dose levels were 0, 25, 125, and 375 ppm flusilazole (96.7% pure). The females were examined daily for evidence of a copulation plug. After the mating period, the females were housed individually. The number of pregnant females/group was 3-6. The total number of pups born did not show a treatment-related effect but two of the five litters at 375 ppm were all dead at birth. The remaining three litters at this dose level were 93-100% viable similar to the litters in the other groups including controls. Mean pup weights at Day 4 were lower in the 375 ppm group than the other three groups. At 375 ppm (equal to 29 mg/kg bw/day) pup weights remained slightly lower than the other groups to weaning (Pastoor et al. 1983). Because of the small group size, the usefulness of this study is limited. Groups of 20 Crl:CD(SD)BR rats/sex/dose level from the chronic toxicity/oncogenicity study were mated within the same dose level for a period of 15 days after a dosing period of about 100 days. Dose levels were 0, 10, 50, and 150 ppm flusilazole (95.6% pure). The females were examined daily throughout the mating period for evidence of a copulatory plug. At the end of the mating period the females were housed individually and allowed to give birth and raise their litters to weaning. About one week after weaning of the F1a litters the females were sacrificed without pathological examination. At weaning of the F1b litters 20 weanlings/sex/group were selected as parents for the F2 generation. These weanlings were maintained on the same diets as their parents for a 90 day period before mating. Mating procedures were the same for the F0 - F1 litters. It is stated that sibling matings were avoided. For each litter the number of pups born, the number of pups born alive, the number alive at 24 hours, 4 days, 12 days and 21 days, the litter weight at 24 hours, and 4 days and the weight of male and female pups at 21 days were recorded. At Day 4 all litters with more than 10 pups were culled to 10, with equal numbers of males and females where possible. Pups found dead were discarded. At weaning of the F2b pups 10/sex/group were randomly selected and necropsied. Liver, kidney, testes and brain were weighed and about 26 tissues/organs were examined for histopathology. Food consumption and body weights were recorded only for the pre-mating periods for the F0 and F1b animals. There were no consistent differences during this period in F0 animals but F1b males given 250 ppm had lower body weights. No differences in food consumption were noted. Pregnancy rates were variable in all mating periods but with no dose-relationship. There was no evidence of a treatment-related effect on male reproductive parameters. There was some indication that gestation length might be longer in the high-dose group but this could not be determined unequivocally because of the number of animals for which gestation lengths could not be determined. The number of pups dead at birth was increased in the 250 ppm group in all litters and in the 50 ppm group in the F2a litter. Survival to Day 4 was reduced at 250 ppm in the F1a, F1b, and F2a litters. Loss of the entire litter perinatally was observed in 2, 2, 2 and 1 dams at 250 ppm in the F1a, F1b, F2a and F2b litters, respectively, and in 1 dam at 50 ppm in the F2a litter. Survival after 4 days was similar in all groups. The sex of pups is indicated only at weaning after culling of the litters so the sex ratio cannot be determined. Pup weights were similar in all groups except for reduced weights at weaning in male and female pups in the F1b and F2a litters. In the F2b weanling pups the absolute and relative liver weights in males in the 250 ppm group were increased. No effect was observed in females. The only pathological lesion of note was hydronephrosis in female weanlings. The incidence of unilateral and bilateral lesions combined was 1/10, 4/10, 3/10, and 5/10 at 0, 10, 50, and 250 ppm, respectively. The incidences in the 10 and 250 ppm groups are slightly higher than those observed in 10 historical control groups over the period 1982-1986. However, neither the severity nor the incidence of the lesions showed a dose-related pattern so it is unlikely that this is treatment-related effect. The NOAEL in this study was 10 ppm (equal to 0.85 mg/kg bw/day on the basis of pre-mating intake data) (Pastoor et al. 1986). Special studies on mutagenicity Flusilazole was negative in seven mutagenicity studies (Table 2). Special studies on teratology Rat In a pilot study in which groups of seven pregnant rats were given doses of 0, 100 or 300 mg flusilazole (99% pure)/kg bw/day Days 7-16 of gestation, 300 mg/kg bw was overtly toxic and 100 mg/kg bw slightly toxic to the dams. Both levels were embryotoxic and 300 mg/kg bw resulted in cleft palates in about 51% of the fetuses in each litter. TABLE 2. RESULTS OF MUTAGENICITY ASSAYS ON FLUSILAZOLE CONCENTRATION TEST SYSTEM TEST ORGANISM OF FLUSILAZOLE RESULTS REFERENCE (PURITY) Ames testa Salmonella typhimurium 1-250 ug/plate Negative Donovan & Irr, 1982 TA1535, TA1537, TA98, (90%) TA100 Ames testa Salmonella typhimurium 5-250 ug/plate Negative Arce et al. 1988 TA1535, TA97, TA98, (97.7%) TA100 Unscheduled DNA Rat hepatocytes Crl (SD) 1 x 10-5 - Negative Chromey et al. 1983 synthesis 1.1 x 102 mM (95.5%) CHO/HGPRT assay Chinese hamster ovary 0.04 - 0.15 mM Negative McCooey et al. 1983 for gene mutationa cells (CHO-K1/BH4 clone) 0.05 - 0.275 mM with and without activation, respectively (95.5%) Micronucleus test Mouse (CD-1) 375 mg/kg bw in Negative Sorg et al. 1984 (bone marrow) corn oil (91.5%) Chromosome aberration Human lymphocytes 1.7 - 100 ug/ml Negative Vlachos et al. 1986 assaya (94.85%) TABLE 2 (CONTD). CONCENTRATION TEST SYSTEM TEST ORGANISM OF FLUSILAZOLE RESULTS REFERENCE (PURITY) Chromosome assay Rat (Crl:CD (SD)) 50 - 500 mg/kg bw in Negative Farrow et al. 1983 (bone marrow) corn oil (not given) a with and without metabolic activation. Groups of 25 female Crl:CD(SD)BR rats mated to males of the same strain were given doses of 0, 10, 50, or 250 mg flusilazole (95.6%)/kg bw/day in corn oil by gavage Days 7-16 of gestation. The day a copulation plug was observed was designated Day 1 of gestation. Body weight, food consumption and clinical condition were monitored throughout the study. On Day 21 of gestation, the rats were killed and examined for gross pathology, liver and uterine weights, and numbers of corpora lutea, implantations, live and dead fetuses. The uterus of any apparently non-pregnant females was stained with ammonium sulfide to detect for early resorptions. The fetuses were weighed and examined for external, visceral and skeletal abnormalities. Chromodacryorrhea, chromorhinorrhea, wet underbodies, or wet or yellow or brown stained perineal areas were observed at the 250 mg/kg bw dose level with all but two females showing at least one of these signs. The severity increased with continued dosing and decreased following cessation of dosing. Some females at this dose level had red vaginal discharges or stains and there was a slightly increased incidence of focal alopecia. Two of the dams died (Days 11 and 18). At 50 mg/kg bw one dam had red vaginal discharge and one ruffled abdominal fur. Food consumption was reduced during dosing at 50 and 250 mg/kg bw. At sacrifice relative liver weights were increased in the dams at 50 and 250 mg/kg bw. At 250 mg/kg bw mean fetal body weight was reduced, incidence of resorptions was increased and the number of live fetuses per litter was reduced. Stunted fetuses were observed only in treated groups at incidences of 1, 4, and 3 fetuses at 10, 50, and 250 mg/kg bw. Cleft palate and absence of renal papilla were observed in fetuses (12% and 9%, respectively) from dams given 250 mg/kg bw but not at lower doses. At 50 mg/kg bw/day two fetuses (from 2 litters) had no innominate artery and one (from the same litter as one of the fetuses with no innominate artery) had a great vessel malformation in the heart. These lesions were not seen in any other group so were probably incidental. In all groups including controls there was a high incidence of hydrocephalus and/or dilated lateral ventricles of the brain. The incidence was not dose-related and no hydrocephalus was observed in subsequent studies so this finding was not considered to be treatment-related. The incidence of misaligned sternebra, extra ossification centres in ribs and ossification delay in sternebra were increased at all dose levels compared to controls. There were increased incidences of rudimentary ribs at 50 and 250 mg/kg bw and extra ribs at 250 mg/kg bw. A NOEL was not demonstrated in this study since skeletal variations were increased at 10 mg/kg bw, the lowest dose tested. Maternal toxicity was seen at 50 mg/kg bw. Teratogenic effects were observed at 250 mg/kg bw (Lamontia et al. 1984a). A second study included groups of 24 mated female Crl:CD(SD)BR rats given doses of 0, 0.4, 2.0, 10.0, 50.0 or 250.0 (10 females) mg flusilazole (95.6%)/kg bw/day in corn oil by gavage Days 7-16 of gestation. The day a copulation plug was observed was designated Day 1 of gestation. Body weight, food consumption and clinical condition were monitored throughout the study. On Day 21 of gestation the rats were killed and examined as in the study above. At 50 and 250 mg/kg bw/day the dams had reduced body weight gains during the early part of the dosing period but the difference was statistically significant only at 250 mg/kg bw. Both groups had reduced food intakes during dosing. Clinical signs of toxicity were observed at 250 mg/kg bw: alopoecia, brown stains of face and limbs and yellow staining of perineum. Relative liver weights were increased in the 50 and 250 mg/kg bw groups. The incidence of early resorptions was similar in all groups; however, the number of dams with median or late resorptions was higher in groups given 10 mg/kg bw or more. The mean number of live fetuses/litter was similar in all groups. The incidence of stunted fetuses was increased at dose levels of 10 mg/kg bw and higher. Cleft palate was observed in fetuses from dams given 250 mg/kg bw but not at lower doses. Absence of renal papilla was observed only in treated groups but there was no dose-relationship and the observation was not considered to be treatment-related. Hydrocephalus was not observed in any group. Visceral (large renal pelvis and small renal papilla) and skeletal (ribs anomalies) variations were observed at 10 mg/kg bw and higher. Absence of the innominate artery was observed in 4(3) fetuses (litters) at 50 mg/kg bw and 1 fetus at 250 mg/kg bw. Retarded development (delayed ossification of sternebra and vertebral arch) was also evident at dose levels of 10 mg/kg bw and higher. The incidence of red foci in bladder of fetuses was increased at dose levels of 10 mg/kg bw and higher; however, these lesions were non-specific and were considered to be of no biological significance. The NOAEL in this study was 2 mg/kg bw/day (Lamontia et al. 1984b). In a dietary study, groups of 24 mated Crl:CD(SD)BR female rats were given diets containing 0, 50, 100, and 300 or 900 ppm flusilazole (94.8%) Days 7-16 of gestation. The day a copulation plug was observed was designated Day 1 of gestation. Body weights, food consumption and clinical condition were monitored throughout the study. On Gestation Day 21 the rats were sacrificed and examined for gross pathology, liver and uterine weights and uterine contents. Fetuses were weighed, sexed and examined for external visceral and skeletal abnormalities. One female at 300 ppm died on Day 19 of gestation. The death was not considered to be related to treatment. During Days 7-11 of gestation at the beginning of dosing weight gains in all treated groups were lower than in controls in a dose-related manner. Significant reductions were noted Days 7-9 at 300 and 900 ppm. Food consumption during dosing was lower than controls in all treated groups but reached significant reductions only at 300 and 900 ppm. There were no treatment-related clinical signs of toxicity. Relative liver weights did not indicate a significant treatment- related effect. Pregnancy rate was high in all groups. One dam at 900 ppm totally resorbed its litter. The incidence of early resorptions was similar in all groups. However, there was an increased incidence (dose-related) of median and late resorptions at 100 ppm and higher. Litter size was inversely dose-related and significantly smaller than in controls in the 900 ppm group. Small litters (less than 10 fetuses) were only seen at dose levels of 100 ppm and above. The 50 ppm group was similar to controls. The incidence of stunted fetuses was increased at 300 and 900 ppm. Mean fetal weights were not affected. No treatment-related malformations were observed. No fetuses showed cleft palate and incidence of absence of renal papilla was not dose or treatment-related. Two fetuses from different litters in the 900 ppm group had slightly domed heads but hydrocephalus was not confirmed on visceral examination. There was a significant dose-related increase in skeletal variations at dose levels of 100 ppm and higher with extra ossification of the sternebra at all 3 dose levels, and rudimentary ribs, extra ossification in cervical ribs and delayed ossification in the vertebral arches (cervical) at 300 and 900 ppm. Red foci were observed in the bladder of a few fetuses at all dose levels with a slightly higher but not dose-related incidence at 100 ppm and above. As in the previous study, these were not considered to be biologically significant. The NOAEL in this study was 50 ppm (equal to 4.6 mg/kg bw) (Alvarez et al. 1984). A study of prenatal and postnatal toxicity in rats was conducted in two phases. For the prenatal study (phase 1), groups of 24 female Crl:CD(SD)BR rats were mated on a 1:1 basis with males of the same strain. The day a copulation plug was observed was designated Day 1 of gestation. On Days 7 through 16 the females were given a dose of 0, 0.2, 0.4, 2, 10, or 100 mg flusilazole (94.8% pure in 0.5% aqueous methyl cellulose)/kg bw by oral intubation. Body weights, food consumption and clinical condition were monitored throughout the study. On Day 21 of gestation the females of one group at each dose level were killed for examination of the uterine contents. The extra control and 100 mg/kg bw groups were killed Day 22 of gestation in order to determine whether absence of renal papillae was a compound-related effect or an anomaly. Analyses of the dosing solutions indicates that the solution prepared for the first day of dosing contained only 6-19% of the nominal concentrations. Since solutions were prepared daily this may have been given to only the one female/group that had reached Gestation Day 7 on that day and only two of these (one at 2 and one at 100 mg/kg) were pregnant. However, the next analyses given were done on solutions prepared six days later and it is unknown whether the technical error causing the low concentrations was corrected before this date. Subsequent analyses indicated concentrations 75-110% of nominal. Assuming the worst case, 13-16 pregnant females/group were given 1-6 doses which were about 10% of the intended dose level (mean duration of low doses was 3.4-3.7 days). In each group there were 7-8 pregnant females which did not receive the low doses. Body weight gains and food consumption were lower than controls during dosing in the females given 100 mg/kg bw. Stained chin was observed in many of the 100 mg/kg bw females and some showed perinasal staining and/or wet perineum. Absolute and relative liver weights were increased in dams at 100 mg/kg bw. Pregnancy rate was 83-96% in treated animals and 96-100% in controls; there was no dose relationship. The number of early resorptions and dams with early resorptions did not differ among the groups. However, there was an increased incidence of median and late resorptions at 100 mg/kg bw. The numbers of stunted fetuses and dams with stunted fetuses were increased at 10 and 100 mg/kg bw (statistically significant only at 100 mg/kg bw). One fetus in each of the 100 mg/kg bw groups had no renal papilla unilaterally. One other high-dose fetus had malformations: no tail and imperforate anus (group killed Gestation Day 22). There was an increased incidence of bladder foci in the two 100 mg/kg bw groups compared to their respective controls. However, the Gestation Day 22 control incidence was higher than the 100 mg/kg bw group killed on Gestation Day 21. Groups given 0.4, 2.0 and 10.0 mg/kg bw had slightly higher incidences of bladder foci. Incidence of small papilla in the kidney was increased at 10 and 100 mg/kg bw but in the groups killed Gestation Day 21 the increase was not dose-related. Incidence of distended ureter was increased at 10 and 100 mg/kg bw when expressed as % affected per litter. The number of litters affected was slightly higher than controls in the Day 21 group at 100 mg/kg bw but not in the Day 22 group. In the postnatal study (phase II), groups of 22 females Crl:CD(SD)BR rats were mated and dosed as in phase I except that only one control and one 100 mg/kg bw group were included. The females were permitted to have their litters and raise them to weaning. Body weights, food consumption and clinical signs were monitored throughout the gestation and lactation periods. Offspring were counted and weighted by sex Days 0, 4 (before and after culling), 7 and 14 post partum. Pups were weighed individually Day 21. At weaning the pups were sacrificed and subjected to gross necropsy. The dosing solutions were adequately prepared in this phase of the study. At 100 mg/kg bw five dams died; four on Gestation Day 24 or 25 (with apparently normal litters) and one four days post partum after delivering a litter (all dead) Day 25. At 2.0 mg/kg bw one dam died Day 24 during delivery. At 100 mg/kg body weight gain and food consumption were reduced during the first few days of dosing but were similar to controls for the remainder of the study. A few dams at 100 mg/kg bw are stated to have shown pallor (4), hunching (2), weakness (2) and dystocia (1) during the puturition and lactation periods. There was an increase in mean gestational length at 10 and 100 mg/kg bw. In controls, gestation did not exceed 22 days. In treated groups a gestation period of 23 days was seen in 2, 1, 4, 12, and 4 dams at 0.2, 0.4, 2.0, 10.0, and 100 mg/kg bw, respectively, and at 100 mg/kg bw 11 dams did not deliver until 24 days gestation. At 100 mg/kg the mean gestation length was 23.7 days compared to 21.8 days in controls. Among the 11 high-dose dams with parturition at Gestation Day 24, six had no live fetuses and the other five had 1-5 live pups (mean 3) and 1-7 dead pups (mean 4.8). At 10 mg/kg bw the longest gestation period was 23 days and there was no apparent effect on pup survival at birth. At this dose four dams had one dead pup (of 6-15 pups in litter) at birth; two delivered Day 22 and two delivered Day 23. This latter incidence of dead pups at birth was similar to that in controls. Survival to Day 4 was decreased at 100 mg/kg bw but was similar to controls at the other dose levels. At 100 mg/kg bw 18% of the pups alive Day 0 of lactation had died by Day 4 compared to 0.4-1.7% of the pups in the control and lower dose groups. There was no effect on survival for the rest of the gestation period. Litter size was reduced at 100 mg/kg bw even when both live and dead pups in the litters were considered. Mean litter size was slightly lower than controls in the 10 mg/kg bw group reaching statistical significance after culling. This was the result of an increased number of litters with less than 10 pups (4 cf 0 in controls and 1-2 in the other treated groups). Pup weights at birth were somewhat higher in the three highest dose groups than in controls. At 2 and 10 mg/kg bw a few litters with higher weights were among those with a 23 day gestation length. At 100 mg/kg bw mean, live litter weights were not as high as at 10 mg/kg bw mean, live litter weights were not as high as at 10 mg/kg bw but other effects confound any comparisons. No treatment-related effects on sex were apparent. The only alterations noted in the weanlings were dilatation of the renal pelvis and distended ureter. The incidence was not unequivocally dose-related although somewhat higher at 10 mg/kg bw/day. It is stated that among the dead pups, absence of renal papilla was observed in only 2/42 pups and small papilla (size 1 or 2) was seen in only 4 pups (2 of each size). Most of the dead pups were in the high-dose group so it is presumed that these observations were from that group. In phase II the NOAEL was 2 mg/kg bw with the only effects being a slightly longer gestation period with a higher mean weight in a few animals (Alvarez et al. 1985a). Rabbit Groups of 18 New Zealand White female rabbits were inseminated with semen from one of the two male rabbits of the same strain from the same source. Day of insemination was designated Gestation Day 0. On Days 7-19 the females were given nominal doses of 0, 2, 5, or 12 mg flusilazole (94.8%)/kg bw/day by gavage in corn oil (actual 1.9, 4.8 and 10.1 mg/kg bw/day). There were two deaths during treatment: one control female Day 19 and one high-dose female Day 16. Another control female died Day 23. Periodic anorexia was observed in some rabbits in all groups and was not related to treatment. No treatment-related effects were noted on clinical observations, body weight, mean food consumption, abortions (2 in controls, 3 in each of the treated groups), number of resorptions, number of live or dead (none in any group) fetuses, fetal weights, maternal liver weights or maternal pathology. The total number of malformed fetuses was not increased by treatment. The incidence of hydrocephalus was 1(1), 2(1), 4(2), and 4(3) fetuses (litters) in the nominal 0, 2, 5, or 12 mg/kg bw/day level groups, respectively. There was no increase in any other malformation nor of any variation. In three subsequent studies hydrocephalus was observed only in one fetus at 35 mg/kg bw/day and in none at any lower dose or in controls. The incidence of hydrocephalus in this study, therefore, was not confirmed and could not be regarded as related to treatment. The NOAEL in this study was the nominal 12 mg/kg bw/day level (actual 10.1 mg/kg bw/day) (Solomon et al. 1984). A second study with 20 New Zealand White female rabbits/group artificially inseminated with sperm from one of three male rabbits tested dose levels of 0, 12, and 35 mg flusilazole (95%)/kg bw/day administered Days 7-19 of gestation by gavage. In this study pregnancy rates were low particularly in the two treated groups (not dose-related). At 35 mg/kg bw/day, 2 of the does aborted and 10 totally resorbed their litters with only one doe with a live litter at examination Day 29. There was an increased incidence of vaginal discharge, discharge in cage and stained tail at this dose level as well as an increased incidence of periodic anorexia. Since there was only one litter, no teratologic effects could be determined. One fetus did have hydrocephalus. Reproductive parameters at 12 mg/kg bw/day did not differ from controls and there was no evidence of maternal toxicity at this level. The NOAEL in this study was the nominal level of 12 mg/kg bw/day (actual 11 mg/kg bw/day) (Zellers et al. 1985). A pilot dietary study in New Zealand White rabbits included 7 females/group offered diets containing 0, 500, 1000, or 2000 ppm flulilazole (94.8%) on Days 7-19 of gestation. Full results are not given but pregnancy rate was low at 500 and 1000 ppm /3/7 at each level and incidence of total resorption was high (4/7) at 2000 ppm. Dietary levels of 0, 300, 600, and 1200 ppm flusilazole (94.8%) were given to groups of 20 New Zealand White female rabbits days 7-19 of gestation following artificial insemination (day of insemination designated Day 0 of gestation). Body weights, food consumption and clinical signs of toxicity were monitored throughout the study. The rabbits were sacrificed Day 29 of gestation and examined for gross pathology, corpora lutea, numbers of live and dead fetuses or resorption sites. The fetuses were examined for external, visceral and skeletal abnormalities. Body weights and food consumption were reduced during dosing. There were no treatment-related clinical signs, pathological lesions or changes in liver weights. Pregnancy rate was reduced in all treated groups with the lowest rate in the high-dose group. The number of dams with total resorptions was increased in the 600 and 1200 ppm groups. There were no treatment-related effects on mean number of live fetuses, resorptions (in dams with fetuses) or fetal weights. The number of litters available at 600 and 1200 ppm was small (3 at each dose level), so the validity of the incidence of fetal malformations and variations is uncertain at these levels. There were no apparent effects at 300 ppm. Since the pregnancy rate was reduced in all treated groups another dietary study was performed with levels of 0, 30, 100, and 300 ppm flusilazole in diet offered Days 7-19 of gestation to groups of 18 or 25 (300 ppm only) female New Zealand White rabbits artificially inseminated with sperm from proven male rabbits. In this study pregnancy rate was low in all groups but particularly in the controls. In the 0 and 300 ppm groups 25% and 29%, respectively, of the pregnant does totally resorbed their litters but there were no total resorptions in the other two groups. Because of the low number of dams with litters available in the control group, the data in this study could not be adequately assessed and a NOAEL cannot be determined (Alvarez et al. 1985b). It has been suggested by the company that the reproductive effects observed in these studies (reduced pregnancy and increased total resorptions) were the result of inadequate technique during artificial insemination. This might explain the effects observed in the control group in the second dietary study. Special studies on skin irritation and sensitization Guinea Pig In young adult Hartley guinea pigs, a range-finding study indicated mild erythema, 24 hours after topical application to the skin of the test material (90% pure flusilazole). The primary irritation study conducted with 5 and 50% solutions in demethyl phthalate did not demonstrate irritation in any of the 10 animals exposed. Intradermal injections of 1% solutions of flusilazole (90% pure) in dimethyl phthalate weekly for 4 weeks in the same animals caused erythema and edema with necrotic centres at the injection sites at 24 hours. Challenge with topical applications of 5 and 50% solutions did not demonstrate sensitization (Wylie et al. 1984d). In another study using 97.7% pure flusilazole, young adult Duncan Hartley albino guinea pits (10/sex) were given 3 weekly dermal applications of the test material (slightly moistened with dimethyl phthalate) which was covered with plastic wrap for 6 hours. No signs of irritation were observed in any of the animals after any of these induction applications. Two weeks after the last induction treatment, the guinea pigs were challenged with a dermal application of test material on an untreated site which was then covered for 6 hours. No reaction was observed in any of the animals. A positive control groups treated with 1-chloro,-2,4-dinitrobenzene (DNCB) showed several erythema with necrosis after the second and third induction applications and severe erythema after the challenge dose two weeks later (Brock et al. 1988). Special study of eye irritation Rabbit Two male New Zealand White rabbits were given doses of 0.01 ml of undiluted flusilazole (90% pure) in the right conjunctival sac. Both rabbits shoed mild conjunctival redness and chemosis at 1-4 hours. The washed eye also showed some discharge at one hour but no other effects. The unwashed eye showed slight corneal opacity at 1-4 hours and slight cloudiness on biomicroscope examination at 24-48 hours. The eye was normal at 72 hours. The test material produced only transient mild irritation (Hall et al. (1984). COMMENTS Flusilazole administered orally to rats was rapidly excreted in urine and feces. The label (14C) from phenyl-labelled material was excreted predominantly in feces, with a sex difference in that males excreted higher amounts in feces than did females. However, substantial variability existed. When triazole-labelled material was given, the 14C was recovered predominantly in urine. Although one study indicated a sex difference, with males excreting more in urine than females, a sex difference was not observed in a second study with more animals. Tissue levels were low. The highest levels were found in the carcass (about 2% of the dose after 96-120 hours). Flusilazole was extensively metabolized in the rat. Recovered parent compound accounted to 2-11% of the dose and was recovered almost exclusively in feces. Flusilazole has a low acute toxicity in the species examined. In short-term (90-day) studies, urothelial hyperplasia in the urinary bladder was observed in rats, mice and dogs at dietary levels of 375, 225, and 125 ppm, respectively. Liver lesions, including hepatocellular hypertrophy and hepatocellular vacuolar cytoplasmic changes, were also observed in the 90-day studies in rats and mice (at 750 and 225 ppm, respectively) and in a one-year study in dogs at 20 ppm. In a one-year feeding study in beagle dogs, centrilobular hepatocyte enlargement was observed at 20 and 75 ppm. The NOAEL was 5 ppm, equal to 0.14 mg/kg bw/day. In long-term feeding studies in rats and mice no bladder lesions were observed. There was urothelial hyperplasia in the renal pelvis in 1 of 10 male rats at 50 ppm and 1 of 10 male rats at 250 ppm for one year, but in none of the rats (7 of 10) given 250 ppm at one year (hepatocellular hypertrophy) and two years (acidophilic foci of cellular alteration: 3/66, 6/63, 4/68, and 13/65 at 0, 10, 50, and 250 ppm, respectively). Male and female mice given 200 ppm for 18 months developed hepatocellular fatty changes (40/80 and 24/80, respectively, versus 4/80 and 2/70 in respective controls). This effect was also reported in 8/80 male mice at 25 ppm, which was not statistically significantly different from controls. There was no evidence of oncogenicity in either species. The NOAEL in the rat study was 10 ppm (equal to 0.45 mg/kg bw/day). The NOAEL in the mouse study was 5 ppm (equal to 0.79 mg/kg bw/day). In a 2-generation, 2 litters per generation reproduction study in rats, flusilazole caused perinatal mortality in both F1 and F2 litters of dams given dietary levels of 250 ppm. There was some indication of increased gestation length at this dose level, an effect which was also observed in a post-natal toxicity study in rats at doses of 10 mg/kg bw/day (gavage) and higher. Perinatal mortality was observed at 100 mg/kg bw/day in this study. The biological significance of prolonged gestation in this study is uncertain. The NOAEL was 10 ppm (equal to 0.895 mg/kg bw/day) but not at lower doses. The NOAEL in rats was 2 mg/kg bw/day. No teratogenic effects were seen in rabbits. The NOAEL was 100 ppm (equal to 2.8 mg/kg bw/day) based on maternal toxicity. After reviewing all available in vitro and in vivo short-term tests, the Meeting concluded that there was no evidence of genotoxicity. TOXICOLOGICAL EVALUATION Level causing no toxicological effect Mouse: 25 ppm in the diet, equal to 3.4 mg/kg bw/day Rat: 10 ppm in the diet, equal to 0.45 mg/kg bw/day Dog: 5 ppm in the diet, equal to 0.14 mg/kg bw/day. Estimate of acceptable daily intake for humans 0-0.001 mg/kg bw. Studies which will provide information valuable in the continued evaluation of the compound 1. Observations in humans. 2. Toxicity of the biotransformation products. REFERENCES Alvarez, L., Krauss, W.C. & Staples, R.E. (1984) Developmental toxicity study in rats given INH-6573-66 in the diet on Days 7-16 of gestation. Unpublished Report No. 432-84 from Haskell Laboratory, Newark, DE, USA. Submitted to WHO by E.I. du Pont de Nemours and Co., Inc., Wilmington, DE, USA. Alvarez, L., Staples, R.E., & Kaplan, A.M. (1985a) INH-6573. Prenatal and postnatal toxicity study in rats dosed by gavage on Days 7-16 of gestation. Unpublished Report No. 654-85 from Haskell Laboratory, Newark, DE, USA. Submitted to WHO by E.I. du Pont de Nemours and Co., Inc., Wilmington, DE, USA. Alvarez, L., Staples, R.E., Driscoll, C.D. & Kaplan, A.M. (1985b) INH-6573. Developmental toxicity study in rabbits treated by diet on Days 7-19 of gestation. Unpublished Report No. 337-85 from Haskell Laboratory, Newark, DE, USA. Submitted to WHO by E.I. du Pont de Nemours and Co., Inc., Wilmington, DE, USA. Anderson, J.J., Stadalius, M.A. & Schlueter, D.D. (1986) Metabolism of 14C-DPX-H6573 in rats. Unpublished Report No. AMR-196-84 from E.I. du Pont de Nemours and Co., Inc., Wilmington, DE, USA. Submitted to WHO by E.I. du Pont de Nemours and Co., Inc., Wilmington, DE, USA. Arce, G.T., Matarese, C.C. & Sarrif, A.M. (1988) Mutagenicity testing of INH-6573-21 in Salmonella typhimurium plate incorporation assay. Unpublished Report No. 59-88 from Haskell Laboratory, Newark, DE, USA. Submitted to WHO by E.I. du Pont de Nemours and Co., Inc., Wilmington, DE, USA. Brock, W.J., Rickard, R.W., Kaplan, A.M. & Gibson, J.R. (1985) Long-term feeding study in mice with INH-6573. Unpublished Report No. 278-85 from Haskell Laboratory, Newark, DE, USA. Submitted to WHO by E.I. du Pont de Nemours and Co., Inc., DE, Wilmington, USA. Brock, W.J., Vick, D.A. & Chromey, N.C. (1988) Closed-patch repeated insult dermal sensitization study (Buehler method) with INH-6573-21 in guinea pigs. Unpublished Report No. 34-88 from Haskell Laboratory, Newark, DE, USA. Submitted to WHO by E.I. du Pont de Nemours and Co., Inc., Wilmington, DE, USA. Cheng, T. (1986) Rat metabolism study of [Triazole-3-14C] DPX-H6573. Unpublished final report 6129-128 from Hazleton Laboratories America, Inc. Submitted to WHO by E.I. du Pont de Nemours and Co., Inc., Wilmington, DE, USA. Chromey, N.C., Horst, A.L., McCooey, K.T. & Sarrif, A.M. (1983) Unscheduled DNA synthesis/rat hepatocytes in vitro. Unpublished Report No. 209-83 from Haskell Laboratory, Newark, DE, USA. Submitted to WHO by E.I. du Pont de Nemours and Co., Inc., Wilmington, DE, USA. Donovan, S.M. & Irr, J.D. (1982) Mutagenicity evaluation in Salmonella typhimurium. Unpublished Report No. 611-82 from Haskell Laboratory, Newark, DE, USA. Submitted to WHO by E.I. du Pont de Nemours and Co., Inc., Wilmington, DE, USA. Farrow, M.G., Cortina, T. & Padilla-Nash, H. (1983) In vivo bone marrow chromosome study in rats - H # 14,728 - Final Report. Unpublished Report No. 201-636 (Haskell No. 288-83) by Hazleton Laboratories America, Inc. Submitted to WHO by E.I. du Pont de Nemours and Co., Inc., Wilmington, DE, USA. Hall, J.A., Dashiell, O.L. & Kennedy, G.L. (1984) Eye irritation test in rabbits. Unpublished Report No. 582-82 from Haskell Laboratory, Newark, DE, USA. Submitted to WHO by E.I. du Pont de Nemours and Co., Inc., Wilmington, DE, USA. Lamontia, C.L., Staples, R.E. & Alvarez, L. (1984a) Embryo-fetal toxicity and teratogenicity study of INH-6573-39 by gavage in the rat. Unpublished Report No. 444-83 from Haskell Laboratory, Newark, DE, USA. Submitted to WHO by E.I. du Pont de Nemours and Co., Inc., Wilmington, DE, USA. Lamontia, C.L., Staples, R.E. & Alvarez, L. (1984b) Embryo-fetal toxicity and teratogenicity study of INH-6573-39 by gavage in the rat. Unpublished Report No. 142-84 from Haskell Laboratory, Newark, DE, USA. Submitted to WHO by E.I. du Pont de Nemours and Co., Inc., Wilmington, DE, USA. McCooey, K.T., Chromey, N.C., Sarrif, A.M. & Hemingway, R.E. (1983) CHO/HGPRT assay for gene mutation. Unpublished Report No. 449-83 from Haskell Laboratory, Newark, DE, USA. Submitted to WHO by E.I. du Pont de Nemours and Co., Inc., Wilmington, DE, USA. O'Neal, F.O., Rickard, R.W., Kaplan, A.M. & Gibson, J.R. (1985) One-year feeding study in dogs with INH-6573. Unpublished Report No. 461-85 from Haskell Laboratory, Newark, DE, USA. Submitted to WHO by E.I. du Pont de Nemours and Co., Inc., Wilmington, DE, USA. Pastoor, T.P., Wood, C.K., Krahn, D.F. & Gibson, J.R. (1983) Ninety-day feeding and one-generation reproduction study in rats with Silane [Bis(4-fluorophenyl)](methyl) (1H-1,2,4-triazol-1- ylmethyl) (INH-6573). Unpublished Report No. 483-83 from Haskell Laboratory, Newark, DE, USA. Submitted to WHO by E.I. du Pont de Nemours and Co., Inc., Wilmington, DE, USA. Pastoor, T.P., Wood, C.K., Drahn, D.F. & Aftosmis, J.G. (1984) Four-week range finding and ninety-day feeding study in mice with Silane [Bis(4-fluorophenyl)] (methyl) (1H-1,2,4-triazol-1-ylmethyl) (INH-6573). Revised Unpublished Report No. 341-83 1-83 from Haskell Laboratory, Newark, DE, USA. Submitted to WHO by E.I. du Pont de Nemours and Co., Inc., Wilmington, DE, USA. Pastoor, T.P., Rickard, R.W., Sykes, G.P., Kaplan, A.M. & Gibson, J.R. (1986) Long-term feeding (combined chronic toxicity/oncogenicity study) and two-generation, four litter reproduction study in rats with INH-6573. Unpublished Report No. 32-86 from Haskell Laboratory, Newark, DE, USA. Submitted to WHO by E.I. du Pont de Nemours and Co., Inc., Wilmington, DE, USA. Poindexter, G.L., Henry, J.E., Kenney, L.A., Burgess, B.A. & Kennedy, G.L. (1984) Inhalation median lethal concentration (LC50) of INH-6573-41 by EPA Protocol. Unpublished Report No. 553-83 by Haskell Laboratory, Newark, DE, USA. Submitted to WHO by E.I. du Pont de Nemours and Co., Inc., Wilmington, DE, USA. Redgate, D., Sarver, J.W. & Chromey, N.C. (1985) Approximate lethal dose (ALD) of INH-6573-66 in rabbits. Unpublished Report No. 54-85 from Haskell Laboratory, Newark, DE, USA. Submitted to WHO by E.I. du Pont de Nemours and Co., Inc., DE, Wilmington, USA. Richard, R.W., Wood, C.K., Krahn, D.F. & Aftosmis, J.G. (1983) Three-month feeding study in dogs with Silane [Bis(4-fluorophenyl)](methyl) (1H-1,2,4-triazol-1-ylmethyl) (INH-6573). Unpublished Report No. 461-83 from Haskell Laboratory, Newark, DE, USA. Submitted to WHO by E.I. du Pont de Nemours and Co., Inc., Wilmington, DE, USA. Sarver, W., Vick, D.A., Valentine, R., Chromey, N.C. & Kaplan, A.M. (1986) Twenty-one dose dermal toxicity study with INH-6573-82 in rabbits. Unpublished Report No. 744-86 from Haskell Laboratory, Newark, DE, USA. Submitted to WHO by E.I. du Pont de Nemours and Co., Inc., Wilmington, DE, USA. Solomon, H.M., Alvarez, L., Staples, R.E. & Hamill, J.C. (1984) Developmental toxicity study in rabbits given INH-6573 by gavage on Days 7-19 of gestation. Unpublished Report No. 333-84 from Haskell Laboratory, Newark, DE, USA. Submitted to WHO by E.I. du Pont de Nemours and Co., Inc., Wilmington, DE, USA. Sorg, R.M., Naismith, R.W. & Mathews, R.J. (1984) Micronucleus test (MNT) OECD H # 15,314. Unpublished Report No. PH 309A-DU-001-84 from Pharmakon Research International (Haskell Report HLO-437-84). Submitted to WHO by E.I. du Pont de Nemours and Co., Inc., Wilmington, DE, USA. Turner, R.J., Kinney, L.A. & Chromey, N.C. (1985) Inhalation median lethal concentration (LC50) of INH-6573 by EPA Guidelines. Unpublished Report No. 1-85 by Haskell Laboratory, Newark, DE, USA. Submitted to WHO by E.I. du Pont de Nemours and Co., Inc., Wilmington, DE, USA. Vlachos, D.A., Covell, D.L. & Sarrif, A.M. (1986) Evaluation of INH-6573-82 in the in vitro assay for chromosome aberrations in human lymphocytes. Unpublished Report No. 745-86 from Haskell Laboratory, Newark, DE, USA. Submitted to WHO by E.I. du Pont de Nemours and Co., Inc., Wilmington, DE, USA. Wylie, C.N., Burgess, B.A. & Kennedy, G.L. (1983) Acute oral test in rats. Unpublished Report No. 78-83 from Haskell Laboratory, Newark, DE, USA. Submitted to WHO by E.I. du Pont de Nemours and Co., Inc., Wilmington, DE, USA. Wylie, C.N., Henry, J.E., Ferenz, R.L., Burgess, B.A. & Kennedy, G.L. (1984a) Median lethal dose (LD50) in rats - EPA proposed guidelines, Newark, DE, USA. Unpublished Report No. 433-83 by Haskell Laboratory, Newark, DE, USA. Submitted to WHO by E.I. du Pont de Nemours and Co., Inc., Wilmington, DE, USA. Wylie, C.N., Henry, J.E., Ferenz, R.L., Burgess, B.A. & Kennedy, G.L. (1984b) Median lethal dose (LD50) in rats - EPA proposed guidelines. Unpublished Report No. 412-83 by Haskell Laboratory, Newark, DE, USA. Submitted to WHO by E.I. du Pont de Nemours and Co., Inc., Wilmington, DE, USA. Wylie, C.N., Henry, J.E., Burgess, B.A. & Kennedy, G.L. (1984c) Ten-dose oral subacute test in rats. Unpublished Report No. 78-83 from Haskell Laboratory, Newark, DE, USA. Submitted to WHO by E.I. du Pont de Nemours and Co., Inc., Wilmington, DE, USA. Wylie, C.N., Henry, J.E., Dashiell, O.L. & Kennedy, G.L. (1984d) Primary skin irritation and sensitization test on guinea pigs. Unpublished Report No. 626-82 from Haskell Laboratory, Newark, DE, USA. Submitted to WHO by E.I. du Pont de Nemours and Co., Inc., Wilmington, DE, USA. Wylie, C.N., Redgate, D., Warheit, D.B. & Chromey, N.C. (1985) Approximate lethal dose (ALD) of INH-6573-66 in male and female mice. Unpublished Report No. 531-84 from Haskell Laboratory, Newark, DE, USA. Submitted to WHO by E.I. du Pont de Nemours and Co., Inc., Wilmington, DE, USA. Zellers, J.E., Staples, R.E., Alvarez, L. & Kaplan, A.M. (1985) Developmental toxicity study (supplemental) in rabbits dosed by gavage on Days 7-19 of gestation. Unpublished Report No. 699-85 from Haskell Laboratory, Newark, DE, USA. Submitted to WHO by E.I. du Pont de Nemours and Co., Inc., Wilmington, DE, USA.
See Also: Toxicological Abbreviations Flusilazole (Pesticide residues in food: 1995 evaluations Part II Toxicological & Environmental)