Regional (RSR) and local species richness (LSR) was recorded for stream invertebrate communities at reference sites in 25 drainage basins in Victoria. Regional species richness was defined as the total number of species recorded at all reference sites within a basin, and LSR as the total numbers of species recorded at a single reference site. Records were obtained from bank and channel habitats and analysed separately. Regressions between LSR and RSR indicated a proportional or linear relationship in both habitats. This applied to the whole data set and to subgroups representing Ephemeroptera, Plecoptera and Trichoptera (EPT as a group), Hemiptera, Diptera, and Coleoptera. All data sets thus represented communities in which no upper limit to LSR was observed. Multiple regressions between LSR and RSR, number of samples per site (N) and seven physical variables showed that RSR and N were nearly always significantly related to LSR. Few of the physical variables were significant except conductivity (for EPT and Coleoptera). Multidimensional scaling ordinations revealed an east-west gradient in compositional similarity of invertebrates, upon which variations in RSR had a major influence. Investigation of factors that regulate RSR will thus be necessary for a broad scale view of what regulates LSR.