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Active environmental DNA ( eDNA ) surveillance through species-specific amplification has shown increased sensitivity in the detection of non-indigenous species ( NIS ) compared to traditional approaches. When many  NIS  are of interest, however, active surveillance decreases in cost- and time-efficiency. Passive surveillance through  eDNA  metabarcoding takes advantage of the complex DNA signal in environmental samples and facilitates the simultaneous detection of multiple species. While passive  eDNA  surveillance has previously detected  NIS , comparative studies are essential to determine the ability of  eDNA  metabarcoding to accurately describe the range of invasion for multiple  NIS  versus alternative approaches. Here, we surveyed twelve sites, covering nine rivers across Belarus for  NIS  with three different techniques, i.e. an ichthyological, hydrobiological and  eDNA  survey, whereby DNA was extracted from 500 ml surface water samples and amplified with two 16S rDNA primer assays targeting the fish and macroinvertebrate biodiversity. Nine non-indigenous fish and ten non-indigenous benthic macroinvertebrates were detected by traditional surveys, while seven  NIS  eDNA  signals were picked up, including four fish, one aquatic and two benthic macroinvertebrates. Passive  eDNA  surveillance extended the range of invasion further north for two invasive fish and identified a new  NIS  for Belarus, the freshwater jellyfish    Craspedacusta   sowerbii   . False-negative detections for the  eDNA  survey might be attributed to: (i) preferential amplification of aquatic over benthic macroinvertebrates from surface water samples and (ii) an incomplete reference database. The evidence provided in this study recommends the implementation of both molecular-based and traditional approaches to maximise the probability of early detection of non-native organisms.

﻿Environmental DNA (eDNA) metabarcoding surveys show evidence of non-indigenous freshwater species invasion to new parts of Eastern Europe

Environmental DNA (eDNA) metabarcoding surveys show evidence of non-indigenous freshwater species invasion to new parts of Eastern Europe