Nationwide Tracing of Two Top Freshwater Fish Invaders in Greece Using Environmental DNA Sampling

Alien fish invasions are causing devastating impacts on native freshwater fauna; thus, rigorous, non-invasive and cost-effective biomonitoring of lotic and lentic freshwaters to design and implement appropriate prevention and control measures is now a priority. In this study, we used a species-specific qPCR eDNA assay to monitor two of the most invasive fish species (Gambusia holbrooki and Pseudorasbora parva) in 15 river basins of Greece and validated these results with conventional fish sampling as well as existing field sampling data. Our monitoring provided new records of invasive species indicating basins for rigorous future monitoring and possible eradication attempts. The eDNA proved more sensitive as a detection tool (56% detection rate) compared to conventional electrofishing (50% detection rate) for G. holbrooki. In contrast, it proved less sensitive for detecting P. parva (38% accuracy) compared to electrofishing (44% accuracy), as evident by the two locations where the eDNA failed to detect the target species. Our study illustrates the potential of the eDNA method for regular, standardised monitoring of riverine habitats for invasive fish species by local managers for early detection. Finally, we discuss the application of eDNA in management interventions for identifying invasive species' hotspots for management prioritisation, for early detection of invaders and for the monitoring of eradication/control actions.