Understanding the relationships of biotic and abiotic factors to seasonal activity at hibernacula is important for the conservation of bats impacted by white-nose syndrome (WNS). Research on the relative and probable activity patterns of the federally endangered northern long-eared bat (Myotis septentrionalis) primarily has focused on summer maternity colonies, whereas surveys at hibernacula have traditionally relied on external capture and internal counts. We used passive acoustic monitoring to assess the relative and probable activity of northern long-eared bats at 13 hibernacula in western Virginia, from August 2020 to May 2022. Northern long-eared bats were most active near hibernacula during warmer weeks of the fall swarm and spring emergence, when rainfall was low. Similarly, the probability of northern long-eared bat activity was highest near hibernacula during the spring/summer season. However, unlike relative activity, the likelihood of recording northern long-eared bats was associated with more heterogeneous, interior forests. Our results suggest that northern long-eared bat activity largely follows the described pre-WNS hibernation phenology of the species. Therefore, acoustical surveys to monitor northern long-eared bat populations at hibernacula should focus on entrances during peak activity periods (mid-April and early September), rather than the nearby landscape. Finally, management to promote resource-rich foraging habitat adjacent to hibernacula for use during swarming and emergence may increase survival during hibernation, fitness for spring migration, and ultimately, improve the reproductive success of northern long-eared bats in western Virginia.