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      Next-generation monitoring of aquatic biodiversity using environmental DNA metabarcoding

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          Abstract

          Global biodiversity in freshwater and the oceans is declining at high rates. Reliable tools for assessing and monitoring aquatic biodiversity, especially for rare and secretive species, are important for efficient and timely management. Recent advances in DNA sequencing have provided a new tool for species detection from DNA present in the environment. In this study, we tested whether an environmental DNA (eDNA) metabarcoding approach, using water samples, can be used for addressing significant questions in ecology and conservation. Two key aquatic vertebrate groups were targeted: amphibians and bony fish. The reliability of this method was cautiously validated in silico, in vitro and in situ. When compared with traditional surveys or historical data, eDNA metabarcoding showed a much better detection probability overall. For amphibians, the detection probability with eDNA metabarcoding was 0.97 (CI = 0.90-0.99) vs. 0.58 (CI = 0.50-0.63) for traditional surveys. For fish, in 89% of the studied sites, the number of taxa detected using the eDNA metabarcoding approach was higher or identical to the number detected using traditional methods. We argue that the proposed DNA-based approach has the potential to become the next-generation tool for ecological studies and standardized biodiversity monitoring in a wide range of aquatic ecosystems.

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              Is Open Access

              MiFish, a set of universal PCR primers for metabarcoding environmental DNA from fishes: detection of more than 230 subtropical marine species

              We developed a set of universal PCR primers (MiFish-U/E) for metabarcoding environmental DNA (eDNA) from fishes. Primers were designed using aligned whole mitochondrial genome (mitogenome) sequences from 880 species, supplemented by partial mitogenome sequences from 160 elasmobranchs (sharks and rays). The primers target a hypervariable region of the 12S rRNA gene (163–185 bp), which contains sufficient information to identify fishes to taxonomic family, genus and species except for some closely related congeners. To test versatility of the primers across a diverse range of fishes, we sampled eDNA from four tanks in the Okinawa Churaumi Aquarium with known species compositions, prepared dual-indexed libraries and performed paired-end sequencing of the region using high-throughput next-generation sequencing technologies. Out of the 180 marine fish species contained in the four tanks with reference sequences in a custom database, we detected 168 species (93.3%) distributed across 59 families and 123 genera. These fishes are not only taxonomically diverse, ranging from sharks and rays to higher teleosts, but are also greatly varied in their ecology, including both pelagic and benthic species living in shallow coastal to deep waters. We also sampled natural seawaters around coral reefs near the aquarium and detected 93 fish species using this approach. Of the 93 species, 64 were not detected in the four aquarium tanks, rendering the total number of species detected to 232 (from 70 families and 152 genera). The metabarcoding approach presented here is non-invasive, more efficient, more cost-effective and more sensitive than the traditional survey methods. It has the potential to serve as an alternative (or complementary) tool for biodiversity monitoring that revolutionizes natural resource management and ecological studies of fish communities on larger spatial and temporal scales.

                Author and article information

                Journal
                Molecular Ecology
                Mol Ecol
                Wiley
                09621083
                February 2016
                February 2016
                January 18 2016
                : 25
                : 4
                : 929-942
                Affiliations
                [1 ]SPYGEN; Savoie Technolac-Bât. Koala 17, Rue du Lac Saint-André-BP 274 Le Bourget-du-Lac Cedex 73375 France
                [2 ]Laboratoire d'Ecologie Alpine (LECA); CNRS; Grenoble 38000 France
                [3 ]Laboratoire d'Ecologie Alpine (LECA); Univ. Grenoble Alpes; Grenoble 38000 France
                [4 ]Laboratoire Biogéographie et Ecologie des Vertébrés; CEFE UMR 5175; Montpellier 34293 France
                [5 ]Hydrosystems and Bioprocesses Research Unit; IRSTEA; Antony Cedex 92761 France
                [6 ]RAVON; Postbus 1413 Nijmegen 6501 BK The Netherlands
                [7 ]Centre for GeoGenetics; Natural History Museum of Denmark; University of Copenhagen; Øster Voldgade Copenhagen Denmark
                [8 ]Direction de l'Action Scientifique et Technique; ONEMA; Vincennes 94300 France
                [9 ]Rhône-Alpes Regional Direction; ONEMA; Bron 69500 France
                [10 ]Centre for Environment, Fisheries and Aquaculture Science; Pakefield Road Lowestoft Suffolk NR33 0HT UK
                [11 ]Environmental and Life Sciences Graduate Program; Trent University; Peterborough ON K9J 7B8 Canada
                [12 ]Pole ONEMA/IRSTEA Hydroécologie des plans d'eau; Centre d'Aix-en-Provence; IRSTEA UR HYAX; Aix-en-Provence 13182 France
                [13 ]Le Sambuc; Tour du Valat; Arles 13200 France
                [14 ]Agence Centre-Ouest; Ecosphère; Orléans 45000 France
                [15 ]LNHE Department; EDF R&D; Chatou Cedex 78401 France
                [16 ]Natural History Museum of Denmark; University of Copenhagen; Universitetsparken 15 Copenhagen 2100 Denmark
                Article
                10.1111/mec.13428
                26479867
                c5db5fa4-1844-464c-8663-8136cb2f38c2
                © 2016

                http://doi.wiley.com/10.1002/tdm_license_1.1

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