Evidence suggests that behavioural defences, such as habitat selection and grooming behaviour, have evolved in animals in response to the costs associated with ectoparasites. Bat fly and mite densities were compared among wild-caught bats in Belize with different roosting preferences (cavity, foliage, or both), and grooming behaviour was analysed for bat species with high and low ectoparasite density. Ectoparasites of bats were removed using forceps, and bat grooming behaviour was recorded with a camcorder. Because bat flies pupate on the surface of host roosts, bats that use cavity roosts (a sheltered environment for the pupae) were predicted to have higher densities of bat flies than those that use foliage (exposed environment). Cavity-roosting species generally had higher densities of bat flies and mites, although the relationship was more evident for bat flies. The grooming behaviour of bats was predicted to differ among species with high or low ectoparasite densities. Although there was no difference in the frequency of grooming behaviours for individuals with and without bat flies, there were differences in grooming behaviour at the species level. Bat species with high ectoparasite densities scratched more than those with low ectoparasite densities. These results suggest that ectoparasite densities and grooming behaviour are related to roosting preferences in bats.