In 2019, dye tracing investigations were conducted near Manitou Cave in Dekalb County, northeast Alabama. The purpose of the dye tracing was to delineate a recharge area for the stream in Manitou Cave, a 1.7-kilometer-long stream cave and the only known habitat for the Manitou Cavesnail (Antroribus breweri). In 2010, the U.S. Fish and Wildlife Service was petitioned by the Center for Biological Diversity to federally list the Manitou Cavesnail. However, before any listing or vulnerability designation can occur, more knowledge was required, specifically regarding potential threats to the snail. With regards to the Manitou Cavesnail, this required delineating a recharge for the stream in Manitou Cave in order to determine what potential threats might exist in the area draining to the cave. From June 2019 to December 2019 two rounds of dye injections, with four locations per round, were conducted to aid in the effort to delineate the recharge area for Manitou Cave. Following this work, a recharge area of 1.22 square kilometers (0.45 square miles) was delineated for the Manitou Cave stream from two positive traces, as well as utilizing information gathered from positive traces to other springs adjacent to Manitou Cave. References Crawford Hydrology Laboratory, 2019, Karst Groundwater Investigation Procedures, Western Kentucky University, Bowling Green, Kentucky, accessed 1/30/2020, https://dyetracing.com/documents/ . Goldscheider, N., Meiman, J., Pronk, M. and Smart, C., 2008. Tracer tests in karst hydrogeology and speleology. International Journal of speleology, 37(1), pp.27-40. Poulain, A., Rochez, G., Van Roy, J.P., Dewaide, L., Hallet, V. and De Sadelaer, G., 2017. A compact field fluorometer and its application to dye tracing in karst environments. Hydrogeology journal, 25(5), pp.1517-1524.