title: Formation of micronuclei in erythrocytes of the fathead minnow (Pimephales promelas) after acute treatment with mitomycin C or cyclophosphamide.
abstract: Despite the widespread use of the fathead minnow in ecotoxicology, there have been relatively few studies on genotoxicity biomarkers in this small, warm-water fish species. Consequently, we investigated the effect of two known genotoxins, mitomycin C and cyclophosphamide, on micronucleus induction in spleen and peripheral blood erythrocytes of this species. Initially, 96-h experiments after intra-peritoneal (i.p.) injections of mitomycin C and cyclophosphamide were undertaken to determine the maximum tolerated dose (MTD). From these studies, MTDs of 10 and 400 mg/kg, respectively, were obtained: doses that were higher than those reported for other fish species. Next, an assessment of micronucleus induction at 1, 2, 4, 8 and 14 days after injection was undertaken for each compound at the MTD. Mitomycin C at 10 mg/kg significantly induced micronuclei in erythrocytes from the spleen, but not from the peripheral blood, at 8 and 14 days. In addition, the overall levels of micronuclei observed were lower than most previously published data from other fish species. In contrast to mitomycin C, treatment with 400 mg/kg cyclophosphamide failed to significantly induce micronuclei in erythrocytes from any of the tissues employed, in contrast to previous reports of significant induction in other species. The reasons for the apparent relative insensitivity of the fathead minnow to these clastogens, with respect to both MTDs and micronucleus induction, are not clear. The fathead minnow, however, has previously been described as relatively insensitive compared to other fish species with respect to selected carcinogens and cytochrome P450 inducers; the latter suggesting that the lack of a significant induction following cyclophosphamide exposure may be due to low metabolic activation in vivo. Consequently, further clarifying work is required to delineate the response shown, considering the extensive use of this species in ecotoxicology research and regulatory testing.
authors.affiliation: AstraZeneca Global Safety, Health and Environment, United Kingdom. matthew.winter@astrazeneca.com
authors.forename: Matthew J
authors.initials: MJ
authors.lastname: Winter
authors.forename: Lucy C J
authors.initials: LC
authors.lastname: Ellis
authors.forename: Thomas H
authors.initials: TH
authors.lastname: Hutchinson
mesh_terms: D000818:Animals; D003520:Cyclophosphamide / Q000008:administration & dosage / Q000633:toxicity*; D003530:Cyprinidae / Q000097:blood / Q000235:genetics*; D004305:Dose-Response Relationship, Drug; D004784:Environmental Monitoring / Q000379:methods; D004912:Erythrocytes / Q000187:drug effects; D008297:Male; D015162:Micronucleus Tests; D016685:Mitomycin / Q000008:administration & dosage / Q000633:toxicity*; D009153:Mutagens / Q000008:administration & dosage / Q000633:toxicity*; D013154:Spleen / Q000187:drug effects
chemical_list: D009153:Mutagens; D016685:Mitomycin; D003520:Cyclophosphamide