Background Understanding the effects of disturbance events, land cover, and weather on wildlife activity is fundamental to wildlife management. Currently, in North America, bats are of high conservation concern due to white-nose syndrome and wind-energy development impact, but the role of fire as a potential additional stressor has received less focus. Although limited, the vast majority of research on bats and fire in the southeastern United States has been conducted during the growing season, thereby creating data gaps for bats in the region relative to overwintering conditions, particularly for non-hibernating species. The longleaf pine (Pinus palustris Mill.) ecosystem is an archetypal fire-mediated ecosystem that has been the focus of landscape-level restoration in the Southeast. Although historically fires predominately occurred during the growing season in these systems, dormant-season fire is more widely utilized for easier application and control as a means of habitat management in the region. To assess the impacts of fire and environmental factors on bat activity on Camp Blanding Joint Training Center (CB) in northern Florida, USA, we deployed 34 acoustic detectors across CB and recorded data from 26 February to 3 April 2019, and from 10 December 2019 to 14 January 2020. Results We identified eight bat species native to the region as present at CB. Bat activity was related to the proximity of mesic habitats as well as the presence of pine or deciduous forest types, depending on species morphology (i.e., body size, wing-loading, and echolocation call frequency). Activity for all bat species was influenced positively by either time since fire or mean fire return interval. Conclusion Overall, our results suggested that fire use provides a diverse landscape pattern at CB that maintains mesic, deciduous habitat within the larger pine forest matrix, thereby supporting the diverse bat community at CB during the dormant season and early spring.Published version