Bat species have differences in diet composition, use of vertical space, and nocturnal activity period. We analyzed diet and partitioning of spatial and temporal resources among fruit bats, and the influence of fruit availability on the number of bat captures in the southern boundary of the Atlantic Forest. Artibeus lituratus and A. fimbriatus fed mainly on Ficus, whereas Sturnira lilium fed on Solanaceae, Piperaceae, and Ficus. However, we did not find correlation between number of fruit bat captures with fruit availability. S. lilium was more active at dusk, whereas A. fimbriatus and A. lituratus were more active at dawn. S. lilium flew more in the understory than canopy and A. lituratus and A. fimbriatus did not show differences in the use of vertical space. There was no difference in the use of vertical space among these fruit bats. We showed that differences in one dimension of the niche, such as food, can help to explain differences in another dimension, such as space use. Niche overlap is greater for related species and studies about niche overlap addressing more than one spatial dimension can elucidate the patterns in use of these resources and also the understanding about bat community composition.