We captured and sampled 68 bats of six species from a shared roosting site in Puerto Rico in April 2012. Bats were screened for Bartonella spp. by culture and confirmed by PCR and sequencing for the gltA gene. Bartonella cultures were obtained from blood specimens of 9/51 (18%) individuals from three species (Artibeus jamaicensis, Brachyphylla cavernarum, and Monophyllus redmani). Phylogenetic analysis of the gltA sequences showed that M. redmani was infected with multiple, diverse Bartonella strains, and A. jamaicensis was infected with a strain related to a strain from a congeneric host. Ectoparasite load could possibly explain observed differences in Bartonella diversity and prevalence between bat species in this community, and we suggest future research to substantiate these preliminary findings.