The distribution and diversity of macroinvertebrates in relation to bison crossings was studied for four seasons in a headwater reach of a tallgrass prairie stream. Species richness was significantly higher above crossings only during the fall. Summer samples revealed significantly higher EPT (richness within only Ephemeroptera, Plecoptera and Trichoptera) index values above crossings than at crossings. On average, over 10% more fine sediment (<0.2 mm) occurred at the crossings than above or below crossings. Densities of two taxa (Hydropsyche spp. and Elmidae) were significantly higher above or below crossings, whereas three taxa (Bezzia/Palpomyia spp., Culicoides sp., and Naididae/Tubificidae) were significantly higher at crossings for at least one season. Overall, this study suggests that impact of bison on benthic communities of headwater streams was spatially limited. Although bison may have less impact upon stream communities than previous studies of cattle impacts have shown, differences in climate and stocking rate could be important factors.