Proper environmental DNA metabarcoding data transformation reveals temporal stability of fish communities in a dendritic river system

Protecting freshwater biodiversity is considered an ultimate challenge but depends on reliable surveys of species distribution and abundance which eDNA metabarcoding (environmental DNA metabarcoding) may offer. To do so, a better understanding of the sources of temporal variation among species eDNA abundance and of data transformation in eDNA metabarcoding studies is needed. Here, we show that transformation based on relative abundance is critical to suitable analyses of eDNA metabarcoding data and that Hellinger transformation performed slightly better than other methods. Furthermore, we show that site localities significantly explain eDNA metabarcoding variation, while no variation is explained by time of sampling. This indicates that species communities vary more spatially than temporally within a dendritic system composed of small rivers. We then further documented the community structure in the St. Charles River (Québec City, Canada) and six of its tributaries. This revealed the existence of eight species communities explaining 82.1% of eDNA read variation within this river network. Moreover, variation in environmental variables among sites explained 53.0% of eDNA reads, while sampling events and temporal environmental variation explained no eDNA metabarcoding variation. Altogether, this supports the claim that eDNA metabarcoding is a powerful tool to document and monitor fish communities in watersheds composed of small river dendritic systems. Protecting freshwater biodiversity is considered an ultimate challenge but depends on reliable surveys of species distribution. Our study supports the claim that eDNA metabarcoding is a powerful tool to document and monitor fish communities in watersheds composed of small river dendritic systems.