Emporia State University (ESU) manages eight natural areas that offer a range of habitats, both aquatic and terrestrial, and are home to a variety of flora and fauna. Throughout the Flint Hills, some fish, turtle, and crayfish species have experienced declines in range and population. To track possible changes in assemblage composition with respect to these animal groups, it is vital to have a baseline understanding of species presence on these areas. The objectives of our study were to compile and organize historical data pertaining to fish, semi-aquatic turtle, and crayfish species' presence on ESU's Natural Areas and to complete a survey of the areas to create an accurate list of the species present on each site. Historical data consisted of field reports from ESU faculty members sent to the Director of the Natural Areas from 1997–present, past master's students' theses, current studies being performed, and communications with faculty members. Surveys were conducted from spring 2018–spring 2019. We found 42 fish, four semi-aquatic turtle, and two crayfish species on the ESU Natural Areas, including new species records on six of the eight areas. Our study created baseline species presence data for the ESU Natural Areas and may serve as a reference for future sampling on these sites.