title: Establishment of the dominant lethal test in the freshwater mollusk Biomphalaria glabrata (Say, 1818).
abstract: Mutagens in the environment may represent a long-term risk for ecosystems. The reproductive potential of populations can be affected by alterations in the fecundity and offspring viability caused by germ cell mutations. Despite the ecological relevance of these effects, there are few studies on germ cell mutagenicity in natural populations. Biomphalaria glabrata was chosen for this study because of the scarcity of data on freshwater invertebrates and the ecological importance of this group. The aim of this study was to establish a germ cell mutagenicity test in B. glabrata by using a similar approach to that used in the dominant lethal test in rodents. Mitomycin C was used as a direct mutagen and cyclophosphamide as a mutagen that requires metabolic activation. Wild-type snails were exposed for 10 days to three concentrations of each agent and crossed with non-exposed albino snails at the end of the treatment. The total frequencies of malformations were analyzed in the offspring of wild-type snails; among the offspring of albino snails, only the heterozygous wild-type embryos were analyzed for malformations. Both agents induced germ cell mutations. The analysis of the offspring of the wild-type snails showed an effect of the exposure up to approximately 5 days after the end of the treatment with cyclophosphamide; the effect of mitomycin C was observed until 45 days after the end of the exposure. There was an increase in the frequencies of malformations in the wild-type offspring of the non-exposed albino snails crossed with the wild-type snails exposed to both agents. The dominant lethal test in B. glabrata proposed in this work is easy to perform, efficient, specific and sensitive in the evaluation of germ cell mutations induced by reference mutagens. The possibility of expanding its use to environmental biomonitoring studies seems very promising and worth trying.
authors.affiliation: Laboratório de Parasitologia/Malacologia, Instituto Butantan, Avenida Vital Brasil, 1500, CEP 05503-900, São Paulo, Brazil. eliananakano@butantan.gov.br
authors.forename: Eliana
authors.initials: E
authors.lastname: Nakano
authors.forename: Liz Cristina
authors.initials: LC
authors.lastname: Watanabe
authors.forename: Fernanda Pires
authors.initials: FP
authors.lastname: Ohlweiler
authors.forename: Carlos Alberto de Bragança
authors.initials: CA
authors.lastname: Pereira
authors.forename: Toshie
authors.initials: T
authors.lastname: Kawano
mesh_terms: D000818:Animals; D001700:Biomphalaria / Q000187:drug effects / Q000196:embryology / Q000235:genetics*; D003433:Crosses, Genetic; D003520:Cyclophosphamide / Q000633:toxicity; D004305:Dose-Response Relationship, Drug; D004625:Embryo, Nonmammalian / Q000002:abnormalities / Q000187:drug effects / Q000473:pathology; D004784:Environmental Monitoring / Q000379:methods*; D005260:Female; D005618:Fresh Water; D005799:Genes, Dominant; D005854:Germ Cells / Q000187:drug effects; D008297:Male; D016685:Mitomycin / Q000633:toxicity; D009152:Mutagenicity Tests / Q000379:methods; D009153:Mutagens / Q000633:toxicity*; D009154:Mutation
chemical_list: D009153:Mutagens; D016685:Mitomycin; D003520:Cyclophosphamide