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ABSTRACT The tricolored bat (Perimyotis subflavus), once common in the eastern United States, has experienced significant mortality due to white‐nose syndrome (WNS), a fungal disease that primarily affects bats hibernating in caves and mines. In coastal regions of the southeastern United States, where caves and mines are scarce, tricolored bats often use roadway culverts as hibernacula. However, WNS infection dynamics in culverts are poorly understood. Previous research indicated that bats with higher body mass at the onset of hibernation have a higher probability of surviving repeated arousal events from WNS. Therefore, we compared tricolored bat winter body mass between cave and culvert hibernacula and identified culvert characteristics influencing body mass during hibernation in Georgia, USA. From 2018 to 2022, we measured body mass of 754 individuals in early and late hibernation across 32 culverts (n = 497) and four caves (n = 257). Our study revealed a southward spread of the fungus over multiple years, with the first confirmed case of WNS in a Georgia culvert in 2022. Overall, tricolored bats in caves weighed more in early hibernation than those in culverts, but bats in culverts weighed more in late hibernation. Across all sites, female tricolored bats entering and leaving hibernation had greater mass than males but lost more mass during hibernation, possibly due to differences in torpor‐arousal patterns and WNS infection rates. Additionally, all bats lost more mass in longer culverts. Understanding culvert characteristics affecting bat body mass will inform management strategies to mitigate WNS effects. Identifying risk factors for specific tricolored bat hibernacula can guide managers on where to focus winter WNS monitoring efforts and potential treatments.

Abstract The potential harm inflicted by forearm bands on bats has been debated for decades. To aid in decision‐making regarding bat marking, we conducted a comprehensive assessment of banding injuries using recapture data from a long‐term overwintering study in Georgia, USA, involving 776 banded tricolored bats (Perimyotis subflavus) with 284 recaptures. Most recaptured bats showed no visible injuries (77.8%); however, 22.2% of bats presented varying degrees of band‐related injuries. Although

Pseudogymnoascus destructans is a fungal pathogen responsible for a deadly disease among North American bats known as white-nose syndrome (WNS). Since detection of WNS in the United States in 2006, its rapid spread and high mortality has challenged development of treatment and prevention methods, a significant objective for wildlife management agencies. In an effort to mitigate precipitous declines in bat populations due to WNS, we have developed and implemented a multi-year mitigation strategy at Black Diamond Tunnel (BDT), Georgia, singly known as one of the most substantial winter colony sites for tricolored bats (Perimyotis subflavus), with pre-WNS abundance exceeding 5000 individuals. Our mitigation approach involved in situ treatment of bats at the colony level through aerosol distribution of antifungal volatile organic compounds (VOCs) that demonstrated an in vitro ability to inhibit P. destructans conidia germination and mycelial growth through contact-independent exposure. The VOCs evaluated have been identified from microbes inhabiting naturally-occurring fungistatic soils and endophytic fungi. These VOCs are of low toxicity to mammals and have been observed to elicit antagonism of P. destructans at low gaseous concentrations. Cumulatively, our observations resolved no detrimental impact on bat behavior or health, yet indicated a potential for attenuation of WNS related declines at BDT and demonstrated the feasibility of this novel disease management approach.