Data are presented on a bat assemblage captured among 10 water holes in Colorado over 5 years. The assemblage consists of Myotis ciliolabrum, M. evotis, M. lucifugus, M. thysanodes, M. volans, Eptesicus fuscus, Lasiurus cinereus, Lasionycteris noctivagans and Corynorhinus townsendii. Results show that reproductive females and juveniles are captured in higher frequencies at water holes containing higher water hardness and that water hardness correlates highly significantly with dissolved calcium content. Also presented are laboratory test data on the stomach volume of Eptesicus fuscus that provide a model for understanding the effect of dissolved calcium content in water as a significant resource. These data indicate that water holes provide supplemental sources of calcium for bats not provided by diet.