The genotoxic effects of three 1,3-butadiene metabolites, 1,2-epoxy-3-butene (monoepoxide, EB), 1,2,3,4-diepoxybutane (diepoxide, DEB) and 1,2-dihydroxy-3,4-epoxybutane (diolepoxide, DiolEB), on male rat germ cells were studied by the meiotic micronucleus method in vitro. Seminiferous tubular segments from stages XII to XIII containing late pachytene-diakinetic spermatocytes were cultivated in the presence of the test chemical for 4 days. During the culture, spermatocytes passed through meiotic divisions and developed into early spermatids in which micronuclei could be scored. DEB was found to be a very potent micronucleus inducer in rat meiosis. All concentrations tested (5-20 microM) were able to cause a statistically significantly higher frequency of micronuclei (P < 0.05) compared with controls and a linear dose-dependent trend for micronucleus induction was seen (P < 0.01). However, EB and DiolEB caused no increase in micronucleus frequencies in spermatids at the concentrations tested (100-1000 microM for EB and 10-100 microM for DiolEB) and at higher concentrations cytotoxic effects were seen upon dividing cells causing a significant reduction in the number of spermatids. According to these results DEB is the most genotoxic butadiene metabolite in rat germ cells during meiosis.