Bats have few natural predators, with birds and snakes being most commonly reported. However, there are also reports of bat predation by large arthropods, including spiders, although recorded cases outside of the Neotropics, Asia, and Australia/Papua New Guinea are rare. Herein, we report an observation of a potential predation event on a Perimyotis subflavus (Tri-colored Bat) by a Dolomedes tenebrosus (Dark Fishing Spider) at a culvert hibernaculum in East Texas. We believe this to be only the second reported case of predation on a bat by a hunting spider in North America. As Tri-colored Bat numbers are decreasing due to the spread of white-nose syndrome, we discuss the need to consider impacts of spider predation at culvert hibernacula and how climatic events may alter the predator–prey dynamics between these 2 species.