Abstract

Freshwater habitats are of high conservation value and provide a wide range of ecosystem services. Effective management requires regular monitoring. However, conventional methods based on direct observation or specimen collection are so invasive, expensive and labour-intensive that frequent monitoring is uncommon. Here, we test whether the evaluation of environmental DNA (eDNA) from water based on a simple protocol can be used for assessing biodiversity. We use universal metazoan primers for characterizing water eDNA across horizontal and vertical spatial dimensions in two reservoirs with known species diversity for two key taxa. eDNA obtained directly from 42 samples × 15 ml water (total = 630 ml) per reservoir yielded DNA signatures for more than 500 metazoan species, of which 105 could be identified to species/genus based on DNA barcodes. We show that eDNA can be used to assign each water sample to its reservoir of origin, and that eDNA outperforms conventional survey methods in single-sample richness comparisons, while revealing evidence for hundreds of unknown species that are undetected by conventional bioassessment methods. eDNA also confirms the presence of a recently discovered invasive snail species and provides evidence for the continued survival of a rare native species of goby not sighted in that habitat since 2007. eDNA thus promises to be a useful addition to the bioassessment toolbox for freshwater systems.

Highlights

  • A relatively small proportion of the Earth’s surface is fresh water, but freshwater habitats are home to a disproportionately large number of species [1] while being extensively used by humans for a wide range of ecosystem services

  • molecular operational taxonomic units (MOTUs) accumulation curves suggest that further sampling would reveal additional MOTUs; they show that MOTU accumulation from surface and benthic samples alone do not differ significantly from the combined accumulation

  • With just 630 ml of water collected from each reservoir in a virtually non-invasive manner within 6 h, environmental DNA (eDNA) metabarcoding detected over 500 metazoan MOTUs using a single primer pair

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Summary

Introduction

A relatively small proportion of the Earth’s surface is fresh water, but freshwater habitats are home to a disproportionately large number of species [1] while being extensively used by humans for a wide range of ecosystem services. Advances in high-throughput DNA sequencing technologies allow for the evaluation of environmental DNA (eDNA) either via metagenomics [2,3] or metabarcoding [4,5,6,7,8] Both techniques rely on detecting species based on trace DNA in the environment. This can be useful for biomonitoring, because obtaining eDNA from fresh water is cost-effective and minimally invasive. Existing studies in the tropics have been largely taxon-specific and/or used filtration prior to DNA extraction [13,18] It remains to be seen whether eDNA samples from different freshwater bodies, sampling sites and sampling depths contain signatures which allow them to be distinguished from one another

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