The southeastern United States is a global hotspot for freshwater aquatic animals. The Conasauga River is one of the most diverse rivers in the Southeast, with 132 native species of fishes, crayfishes, and mussels, ranking 21st out of 290 Southeastern watersheds for conservation priority. While the majority of the Conasauga watershed is in Georgia, the upper Conasauga River proper and several of its tributaries are in Polk and Bradley counties, Tennessee. There are four are federally listed fishes (of 83 total native species) in the entire Conasauga watershed, including two threatened species, the Blue Shiner (Cyprinella caerulea) and the Trispot Darter (Etheostoma trisella). We surveyed for Blue Shiners throughout their range in the upper Coosa River system in Alabama. Georgia, and Tennessee from 2017-2019. Blue Shiners were present at 21 of 27 sites sampled. Most of the sites where they were absent were tributaries with no records of Blue Shiners, such as Sugar and Mill creeks in Tennessee. Blue Shiners were found at all sites in the Conasauga River proper and Minnewauga Creek in Tennessee, and were collected in good numbers with several size classes. Although not targeted, we also collected Trispot Darters at several sites in the Holly Creek watershed in Georgia. With Georgia Department of Natural Resources, we are currently ground-truthing positive hits from environmental DNA in the Conasauga River watershed for Trispot Darters where no previous records exist. We are also searching for new breeding sites for this species, which are small, ephemeral tributaries with groundwater input .