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Interactions of bats with their roosts have played an important role in bat evolution. These interactions are well-known in frugivorous bats, where it has been found that several morphological characters may be associated to specific roosts, as in tent-roosting species from subtribes Vampyressina and Artibeina or foliage-roosting bats Stenodermatina. However, data on interactions between bats and roosts are scarce for several other groups. This is the case for Lophostoma species and termite nests, which are the preferred shelters for this genus. Herein we provide new data on roosting ecology of the bat Lophostoma brasiliense, along with information on diet and activity patterns. We additionally compiled data on use of termite nests by these bats. L. brasiliense consumed 11 food items, mainly Coleoptera and Heterocera and were more active between 00: 00 and 02: 00 h. According to our results L. brasiliense and L. silvicolum are strongly associated with termite nests and each of these species appears to have particular roost requirements. The use of termite nest is currently documented for 15 bat species with a high frequency in Lophostoma species, but evolutionary mechanisms that generated the use of this type shelter remain unknown.