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Kgalagadi Transfrontier Park is located in northwestern South Africa and extends northeastward into Botswana. The park lies largely within the southern Kalahari Desert ecosystem where the Auob and Nassob rivers reach their confluence. Although these rivers run only about once every 100 years, or shortly after large thunderstorms, underground flows and seeps provide consistent surface water for the parks sparse vegetation and diverse wildlife. No formal studies on bats have previously occurred at Kgalagadi. We used SM2 + BAT ultrasonic detectors to survey 10 sites along the Auob and Nassob rivers from 5-16 April 2016. The units recorded 3960 call sequences that were analyzed using Kaleidoscope software for South African bats as well as visual determinations based on call structure attributes (low frequency, characteristic frequency, call duration, and bandwidth). We identified 12 species from four families: Rhinolophidae: Rhinolophus fumigatus. Molossidae: Chaerephon pumilus, and Sauromys petrophilus, Tadarida aegyptiaca; Miniopteridae: Miniopteris schreibersi (natalensis), Vespertilionidae: Laephotis botswanae, Myotis tricolor, Neoromicia capensis, N. nana, Pipistrellus hesperidus, Scotophilus dinganii, and S. viridus. The most abundant species during the survey period was N. capensis. We also used paired-site design to test for greater bat activity at water sources compared to dry sites, with dry sites being significantly more active. We conclude that species richness is much higher than previously known from this region and that more species may be present during the warmer months of the year. In addition, activity of bats during the dry season in Kgalagadi would likely be more concentrated around drinking opportunities, thus allowing for better detection of species richness in the area.