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      Targeted and passive environmental DNA approaches outperform established methods for detection of quagga mussels, Dreissena rostriformis bugensis in flowing water

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          Abstract

          1. The early detection of invasive non‐native species (INNS) is important for informing management actions. Established monitoring methods require the collection or observation of specimens, which is unlikely at the beginning of an invasion when densities are likely to be low. Environmental DNA (eDNA) analysis is a highly promising technique for the detection of INNS—particularly during the early stages of an invasion.

          2. Here, we compared the use of traditional kick‐net sampling with two eDNA approaches (targeted detection using both conventional and quantitative PCR and passive detection via metabarcoding with conserved primers) for detection of quagga mussel, Dreissena rostriformis bugensis, a high priority INNS, along a density gradient on the River Wraysbury, UK.

          3. All three molecular tools outperformed traditional sampling in terms of detection. Conventional PCR and qPCR both had 100% detection rate in all samples and outperformed metabarcoding when the target species was at low densities. Additionally, quagga mussel DNA copy number (qPCR) and relative read count (metabarcoding) were significantly influenced by both mussel density and distance from source population, with distance being the most significant predictor.

          4. Synthesis and application. All three molecular approaches were more sensitive than traditional kick‐net sampling for the detection of the quagga mussel in flowing water, and both qPCR and metabarcoding enabled estimates of relative abundance. Targeted approaches were more sensitive than metabarcoding, but metabarcoding has the advantage of providing information on the wider community and consequently the impacts of INNS.

          Abstract

          This study demonstrates the increased sensitivity and ability of detecting INNS in rivers using three different eDNA methods (conventional PCR, qPCR, and a metabarcoding) compared to traditional kick‐net sampling. Furthermore, we validate our approaches with mesocosm and field experiments.

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          Environmental DNA metabarcoding: Transforming how we survey animal and plant communities

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            A new versatile primer set targeting a short fragment of the mitochondrial COI region for metabarcoding metazoan diversity: application for characterizing coral reef fish gut contents

            Introduction The PCR-based analysis of homologous genes has become one of the most powerful approaches for species detection and identification, particularly with the recent availability of Next Generation Sequencing platforms (NGS) making it possible to identify species composition from a broad range of environmental samples. Identifying species from these samples relies on the ability to match sequences with reference barcodes for taxonomic identification. Unfortunately, most studies of environmental samples have targeted ribosomal markers, despite the fact that the mitochondrial Cytochrome c Oxidase subunit I gene (COI) is by far the most widely available sequence region in public reference libraries. This is largely because the available versatile (“universal”) COI primers target the 658 barcoding region, whose size is considered too large for many NGS applications. Moreover, traditional barcoding primers are known to be poorly conserved across some taxonomic groups. Results We first design a new PCR primer within the highly variable mitochondrial COI region, the “mlCOIintF” primer. We then show that this newly designed forward primer combined with the “jgHCO2198” reverse primer to target a 313 bp fragment performs well across metazoan diversity, with higher success rates than versatile primer sets traditionally used for DNA barcoding (i.e. LCO1490/HCO2198). Finally, we demonstrate how the shorter COI fragment coupled with an efficient bioinformatics pipeline can be used to characterize species diversity from environmental samples by pyrosequencing. We examine the gut contents of three species of planktivorous and benthivorous coral reef fish (family: Apogonidae and Holocentridae). After the removal of dubious COI sequences, we obtained a total of 334 prey Operational Taxonomic Units (OTUs) belonging to 14 phyla from 16 fish guts. Of these, 52.5% matched a reference barcode (>98% sequence similarity) and an additional 32% could be assigned to a higher taxonomic level using Bayesian assignment. Conclusions The molecular analysis of gut contents targeting the 313 COI fragment using the newly designed mlCOIintF primer in combination with the jgHCO2198 primer offers enormous promise for metazoan metabarcoding studies. We believe that this primer set will be a valuable asset for a range of applications from large-scale biodiversity assessments to food web studies.
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              Update on the environmental and economic costs associated with alien-invasive species in the United States

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                Author and article information

                Contributors
                Rosetta.Blackman@eawag.com
                Journal
                Ecol Evol
                Ecol Evol
                10.1002/(ISSN)2045-7758
                ECE3
                Ecology and Evolution
                John Wiley and Sons Inc. (Hoboken )
                2045-7758
                29 October 2020
                December 2020
                : 10
                : 23 ( doiID: 10.1002/ece3.v10.23 )
                : 13248-13259
                Affiliations
                [ 1 ] Department of Aquatic Ecology Eawag Swiss Federal Institute of Aquatic Science and Technology Dübendorf Switzerland
                [ 2 ] Department of Evolutionary Biology and Environmental Studies University of Zurich Zürich Switzerland
                [ 3 ] Evolutionary and Environmental Genomics Group (EvoHull) Department of Biological and Marine Sciences University of Hull Hull UK
                [ 4 ] School of Biological and Environmental Sciences Liverpool John Moores University Liverpool UK
                Author notes
                [*] [* ] Correspondence

                Rosetta Blackman, Department of Aquatic Ecology, Eawag, Swiss Federal Institute of Aquatic Science and Technology, Überlandstrasse 133, CH‐8600 Dübendorf, Switzerland.

                Email: Rosetta.Blackman@ 123456eawag.com

                Author information
                https://orcid.org/0000-0002-6182-8691
                https://orcid.org/0000-0003-0923-1801
                https://orcid.org/0000-0001-8154-9828
                https://orcid.org/0000-0001-7630-9360
                https://orcid.org/0000-0002-8153-5511
                Article
                ECE36921
                10.1002/ece3.6921
                7713958
                33304534
                be255209-92e3-44a2-841d-46b91a4e65ff
                © 2020 The Authors. Ecology and Evolution published by John Wiley & Sons Ltd.

                This is an open access article under the terms of the http://creativecommons.org/licenses/by/4.0/ License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.

                History
                : 17 January 2020
                : 14 September 2020
                Page count
                Figures: 4, Tables: 1, Pages: 12, Words: 10244
                Funding
                Funded by: Gilson Le Cren Award
                Categories
                Original Research
                Original Research
                Custom metadata
                2.0
                December 2020
                Converter:WILEY_ML3GV2_TO_JATSPMC version:5.9.5 mode:remove_FC converted:03.12.2020

                Evolutionary Biology
                early detection,edna,invasive non‐native species,metabarcoding,pcr,qpcr,targeted assay

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