Thirteen species of bats occur in Nebraska, but limited information is available on those inhabiting southwestern parts of the state. We investigated the distribution, abundance, and reproductive activity of bats in 5 counties in extreme southwestern Nebraska (Chase, Hays, Dundy, Hitchcock, and Red Willow). From April 2007 to April 2008, we deployed mist nets on 15 occasions at 8 localities over the Republican River and its tributaries. We captured 100 individuals representing 5 species, including the evening bat (Nycticeius humeralis), eastern red bat (Lasiurus borealis), hoary bat (Lasiurus cinereus), big brown bat (Eptesicus fuscus), and silver-haired bat (Lasionycteris noctivagans). All species raised young in this region of Nebraska, except L. noctivagans, which was documented only during migration. Lactating females of N. humeralis captured on 15 June and volant young captured on 23 June extend known dates of reproductive activity for this species in the state, and an adult female captured on 30 April represents the earliest seasonal record of this species from Nebraska. Our records of evening bats also extend the known distribution of this species farther west in Nebraska than previously reported. Changes in land use throughout the Great Plains during the last century have altered many habitats, such as gallery forests associated with rivers, and distributions of mammals, including bats, have shifted to reflect those ecological changes.