The southern brown bandicoot (Isoodon obesulus) has undergone significant range contractions since European settlement, and it is now considered ''Endangered'' throughout south-eastern mainland Australia. This species currently has a highly fragmented distribution inhabiting a mosaic of habitats. This project uses mitochondrial DNA (mtDNA) and microsatellite data to determine levels of genetic diversity, population structure and evolutionary history, which can aid wildlife managers in setting priorities and determining management strategies. Analyses of genetic diversity revealed low levels of mtDNA variability (mean h ¼ 50.42%, p ¼ 0.76%) and divergence (mean d A ¼ 0.29%) across all regions investigated, and was among the lowest recorded for marsupials. These data indicate a relatively small female effective population size, which is most likely a consequence of a large-scale population contraction and subsequent expansion occurring in pre-history (mismatch distribution analysis, SSD P-value ¼ 0.12). Individuals from the Sydney region experienced significant reductions in microsatellite diversity (A ¼ 3.8, H E ¼ 0.565), with the Garigal National Park (NP) population exhibiting ''genetic reduction signatures'' indicating a recent population bottleneck. Population differentiation analysis revealed significant genetic division amongst I. obesulus individuals from Sydney, East Gippsland and Mt Gambier regions (h ¼ 0.176-0.271), but could not separate the two Sydney populations (Ku-ring-gai NP and Garigal NP). Based on these data and habitat type, translocations could readily be made between the two Sydney populations, but not between the others. Phylogenetic comparisons between I. obesulus and I. auratus show little support for current Isoodon taxonomy, consistent with the findings of Pope et al. 2001. We therefore recommend the recognition of only three I. obesulus sub-species and suggest that these comprise a single morphologically diverse species that once was widespread across Australia.