The Ecology Section of the Georgia Department of Transportation (GDOT) is charged with the assessment of ecological impacts of proposed transportation projects, including potential impacts to protected species. In Georgia, the Apalachicola-Chattahoochee-Flint River (ACF) Basin is home to four freshwater mussel species listed as endangered and two species listed as threatened under the Endangered Species Act. Traditional surveys for rare mussel species can be time consuming and sometimes ineffective due to the cryptic nature of these animals. The application of environmental DNA (eDNA; DNA extracted from environmental samples) metabarcoding for conducting mussel surveys has several advantages over traditional methods including relatively simple and non-invasive field methods and potential for improved detection sensitivity. eDNA detection of rare freshwater mussels may be limited by the volume of water that can be filtered. Dead-end ultrafiltration (DEUF) is a water filtration method that enables the filtration of large volumes of water. DEUF is commonly used to sample environmental waters for pathogens; however, DEUF has not been tested in eDNA studies for metazoan taxa. The United States Geologic Survey (USGS) has partnered with GDOT to conduct a pilot study in the summer of 2021 to test recently developed mussel metabarcoding markers on eDNA samples collected with DEUF in the ACF Basin at sites with known high diversity of freshwater mussel species. Anticipated outcomes of this work include a qualitative assessment of eDNA as an option for conducting mussel community surveys and a qualitative comparison of mussel detection probabilities between eDNA samples collected with standard filters and DEUF. If evidence suggests threatened and endangered mussels in the ACF Basin can be consistently detected with eDNA metabarcoding, further research would be warranted to statistically compare results between traditional and eDNA mussel surveys to determine conditions under which eDNA may be used as a supplement to or replacement of traditional survey methods. Additionally, results from testing the efficacy of eDNA detections with DEUF versus standard filters can inform future eDNA surveys for freshwater mussel species. This presentation will focus on the results of the study, discuss potential benefits to species and GDOT project delivery, and outline a framework for others who may want to pursue or adapt this new approach.