The effects of urbanization on biodiversity are generally considered to be negative, but the potential for landscape context to modulate these effects has not been adequately examined because most urban ecology research has been conducted in one biome: the temperate forest. This bias also applies to studies of the urban ecology of bats, whose diversity is correlated with habitat heterogeneity. We investigated the hypothesis that in the fairly flat, homogeneous Prairies, urbanization, by creating structurally complex islands, benefits bats by increasing access to the vertical landscape elements (buildings and trees) in which they roost. From 2006 to 2008, we surveyed bat assemblages in and around Calgary, Alberta, using mist nets to capture them and bat detectors to record their echolocation activity. Our data supported the prediction that urbanization increases the abundance of Prairie bats, but not the prediction that it increases their diversity. Instead, the urban bat assemblage was less diverse, and exhibited decreased species evenness compared to non-urban assemblages. Although Myotis lucifugus dominated bat assemblages throughout our study area, this was most evident in the city, and this species drove the increased urban abundance of bats. Ultimately, we reject our hypothesis and conclude that urbanization in the Prairies may create attractive habitat for one synanthropic bat, but is detrimental to others.