eDNA detection of autochthonous and invasive freshwater crayfish in Trentino

A major threat to biodiversity conservation is posed by invasive alien species (IAS). The global human-driven spread of these organisms reduces autochthonous species populations through resource competition and introduction of novel pathogens. Fast and efficient detection of invasive species is of fundamental importance for controlling their diffusion, and environmental DNA (eDNA) analysis is being developed to provide a cheaper and less time-consuming tool for early-detection. Our present aim was to test and apply a new eDNA protocol for monitoring white-clawed crayfish (Austropotamobius pallipes) populations in the Province of Trento, as well as the two invasive species known to be present in the Province: the red-swamp crayfish (Procambarus clarkii) and the spiny-cheek crayfish (Faxonius limosus). After filtering water samples from 14 sites across the study area, we extracted whole DNA using the DNeasy PowerWater Kit, and amplified a 360bp COI fragment with a single multi-species primer pair that allows discrimination of the three crayfish species, as well as A. pallipes mtDNA haplotypes. PCR products were sequenced with Illumina paired-end technology. Here we discuss the success of our method and present our preliminary results on freshwater crayfish detection using eDNA.