Environmental DNA and Karst Systems: Advancing Groundwater Tracing, Aquifer Connectivity, and Ecosystem Research – Mike Buchanan 2025 Abstract Karst systems are complex dynamic environments that present significant challenges to hydrological and ecological research. Traditional methods, such as dye tracing, provide limited spatial and temporal resolution in groundwater studies. Recent advances in environmental DNA (eDNA) analysis have opened new frontiers, offering powerful, non-invasive tools for exploring species distribution, ecosystem functioning, groundwater flow, and aquifer connectivity. This paper synthesises current knowledge on the use of eDNA in karstological research and groundwater tracing, highlighting its advantages, limitations, and future research needs. By integrating both ecological and hydrological applications, this review demonstrates the transformative potential of eDNA in advancing karst science. Introduction Karst systems represent some of the mos...