Phylogeographic analysis of long-legged bats, <i>Macrophyllum macrophyllum</i>, with notes on roosting behavior and natural history

The long-legged bat (<i>Macrophyllum macrophyllum</i>) is widely distributed in the continental Neotropics, but poorly known because it is not commonly caught in mist nets. Available data suggest that this species is closely associated with water where it forages for insect prey. We compiled the first comprehensive molecular dataset assembled for the species, spanning its entire distributional range to investigate if the phylogeography of this monotypic genus is associated with the hydrographic drainage, ecosystem regions, or genetic clustering in Central and South America. To survey under sampled areas, fieldwork was conducted in the Brazilian Pantanal and Cerrado targeting the search for riverine roost sites of <i>Macrophyllum</i>. A literature review was also done to summarize roosting information for the species. New sequences of the mitochondrial cytochrome <i>b</i> gene were generated for tissue samples from Brazil and in museum collections. Phylogenetic trees were constructed using both maximum likelihood and Bayesian inference methods and a haplotype network was used to analyze population structure. Our phylogenetic results identified five geographic lineages of <i>Macrophyllum</i> from (1) the western Cerrado, (2) eastern Cerrado and Pantanal, (3) Guianas, (4) Amazonia, and (5) Central America. However, the haplotype network in conjunction with the genetic clustering identified four populations with the eastern Cerrado and Pantanal grouping with the Guianas and the eastern part of Amazonia. The fieldwork in the Cerrado and Pantanal along with the literature review identified that about half of the roost sites for the long-legged bats were drainage culverts. There is geographic structuring in the mitochondrial data of <i>Macrophyllum</i> with Central America, western Cerrado, Pantanal, Guianas, and eastern Ecuador reciprocally monophyletic and well differentiated populations. However, the under sampled eastern Amazonia is poorly resolved in relation to the other areas. The long-legged bats seem to be relatively adaptable to certain levels of human disturbance and landscape development with man-made drainage culverts commonly used as roosting sites. Increased biodiversity surveys of bats in central Brazil are needed to fill in distributional gaps, such as the lower Amazon River basin, to resolve phylogeographic patterns of <i>Macrophyllum</i> in South America and better understand the potential of cryptic species in this monotypic genus.