Abstract: We assessed parentage within and among maternity colonies of the northern long-eared bat (Myotis septentrionalis Troessart 1897) in north-central Kentucky from 2011–2013 to better understand colony social structure, formation, and membership dynamics. We intensively sampled colonies in close and remote (> 10 km) spatial proximity both before and after targeted day-roost removal. Colonies were not necessarily comprised of closely related individuals, but natal philopatry was common. Adjacent colonies often contained maternally related individuals, indicating that some pups did disperse, albeit not far from their natal home range. Lack of apparent overlap among maternity colonies, along with no observed individual movements between colonies, suggests that colonies may be relatively closed once established in the maternity season. Whereas some young on site had been sired by males collected on site that by chance had dispersed to the same summering grounds, most had not, as would be expected since the species mates in the fall swarms near hibernacula. The number of parentages that we inferred among colonies, however, suggests that outside the maternity season, social groups may be relatively flexible and open, with individuals moving among groups close to their natal area. Analysis of microsatellite DNA data showed a low FST (= 0.011) and best fit to a model of one multilocus genotypic cluster across the study area. We observed high turnover in colony membership between years in all colonies, regardless of roost removal treatment. Our results suggest that female northern long-eared bats exhibit fidelity to a general geographic area rather than individual colonies between years, and indicate presence of a complex and dynamic social-genetic structure. Greater understanding of colony dynamics, including formation, dissolution, and dispersal patterns, may contribute to conservation and management of this threatened species.