Pseudogymnoascus destructans, the causal agent of white-nose syndrome (WNS), is commonly found on bats captured both inside and outside caves during hibernation, a time when bats are most vulnerable to infection. It has not been documented in the southeast US on bats captured outside caves or on the landscape in summer. We collected 136 skin swabs from 10 species of bats captured at 20 sites on the Tennessee side of Great Smoky Mountains National Park, 12 May–16 August 2015. Three swabs were found positive for P. destructans, one from a male tricolored bat (Perimyotis subflavus) and two from male big brown bats (Eptesicus fuscus). This detection of P. destructans on free-flying male bats in the southeast US during summer has potential repercussions for the spread of the fungus to novel bat species and environments. Our finding emphasizes the need to maintain rigorous year-round decontamination of field clothing and equipment until more is understood about the viability of P. destructans found on bats captured outside hibernacula during summer, about the potential for males to act as reservoirs of the fungus, and the risk of fungal transmission and spread.