Development and validation of eDNA markers for the detection of Crepidula fornicata in environmental samples

The invasive Crepidula fornicata caused major problems along the European Atlantic coast, especially in France and Netherlands where high densities leads on changes in the habitat, disturb native marine wildlife as well as it originates competition for space and food. Despite its dangerous invasive nature, regular monitoring to alert about its presence in risk areas, like the south Bay of Biscay (Spain and south France), is not done yet. Here, we developed a species-specific marker to detect the presence of C. fornicata in environmental samples (eDNA) of seawater. The novel C. fornicata specific primers amplified a region of 239 bp within the COI gen. We employed this tool to check its presence in 6 estuaries of the Cantabrian Sea, an area comprised between the Spanish and French limits of the previously reported presence of this limpet in the south Bay of Biscay. The presence of C. fornicata was confirmed in A Coruña (Galicia, Spain), Eo and Villaviciosa estuaries (Asturias, Spain) while it was not detected in Santander, Bilbao (Spain), and Bayonne (France). This new method to detect C. fornicata could be easily implemented in regular monitoring to prevent and manage future invasions of this species.