Alpine freshwater fish biodiversity assessment: an intercalibration test for metabarcoding method set up

Environmental DNA (eDNA) based methods (Fig. 1) are proving to be a promising tool for freshwater fish biodiversity assessment in Europe within the Water Framework Directive (WFD, 2000/60/EC) especially large rivers and lakes where current monitoring techniques have known shortcomings. Freshwater actively involved aquatic ecosystems functioning diversity, contributing health, well-being economy every geographic realm. Unfortunately, many experiencing critical population decline with risk of local or global extinction because intense anthropogenic pressure. Within EU project Eco-AlpsWater, advanced high throughput sequencing (HTS) used improve traditional WFD approaches by using environmental collected Alpine waterbodies. To evaluate performance metabarcoding approach specifically designed measure rivers, an intercalibration test was performed. This exercise forecasted use mock samples containing either tissue-extracted different target species water from aquaculture tanks mimic real sampling processing. Moreover, three Lake Bourget (France) were compare efficiency taxonomic assignments natural community samples. Our results highlighted good molecular laboratory protocols HTS amplification success selected primers, providing essential information concerning resolution 12S mitochondrial marker. As further confirmation, concentration well represented relative read abundance. preliminary confirmed applicability eDNA analyses biomonitoring inhabiting perialpine rivers.