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Crayfish of North American origin are amongst the most prominent high-impact invasive invertebrates in European freshwaters. They contribute to the decline of European native crayfish species by spreading the pathogen causing crayfish plague, the oomycete    Aphanomyces   astaci   . In this study we validated the specificity of four quantitative PCR ( qPCR ) assays, either published or newly developed, usable for environmental DNA ( eDNA ) screening for widely distributed native and non-native crayfish present in Central Europe:    Astacus   astacus   ,    Pacifastacus   leniusculus   ,    Faxonius   limosus    and    Procambarus   virginalis   . We then conducted an  eDNA  monitoring survey of these crayfish as well as the crayfish plague pathogen in a wide variety of habitat types representative for Central and Western Europe. The specificity of  qPCR  assays was validated against an extensive collection of crayfish DNA isolates, containing most crayfish species documented from European waters. The three assays developed in this study were sufficiently species-specific, but the published assay for    F.   limosus    displayed a weak cross-reaction with multiple other crayfish species of the family   Cambaridae  . In the field study, we infrequently detected  eDNA  of    A.   astaci    together with the three non-native crayfish species under examination. We never detected  eDNA  from    A.   astaci    together with native crayfish, but in a few locations  eDNA  from both native and non-native crayfish was captured, due either to passive transport of  eDNA  from upstream populations or co-existence in the absence of infected crayfish carriers of    A.   astaci   . In the study, we evaluated a robust, easy-to-use and low-cost version of the  eDNA  sampling equipment, based mostly on items readily available in garden stores and hobby markets, for filtering relatively large (~5 l) water samples. It performed just as well as the far more expensive equipment industrially designed for  eDNA  water sampling, thus opening the possibility of collecting suitable  eDNA  samples to a wide range of stakeholders. Overall, our study confirms that  eDNA -based screening for crayfish and their associated pathogen is a feasible alternative to traditional monitoring.

neobiota

Simultaneous detection of native and invasive crayfish and    Aphanomyces   astaci    from environmental DNA samples in a wide range of habitats in Central Europe