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      DNA metabarcoding reveals metacommunity dynamics in a threatened boreal wetland wilderness

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          Significance

          Too often, ecological monitoring studies are designed without understanding whether they have sufficient statistical power to detect changes beyond natural variability. The Peace–Athabasca Delta is North America’s largest inland delta, within a World Heritage area, and is currently threatened by human development. Using multispecies occupancy models we show that the wetland macroinvertebrate community is highly diverse, and spatial and temporal turnover are so high that composition is nearly random, emphasizing stochastic processes of assembly. Using DNA metabarcoding, our study detected more taxa, both overall and per sample, than traditional morphology-based sample processing, increasing our power to detect ecosystem change. Improving data quality and quantifying error are key to delivering effective monitoring and understanding the dynamic structure of the metacommunity.

          Abstract

          The complexity and natural variability of ecosystems present a challenge for reliable detection of change due to anthropogenic influences. This issue is exacerbated by necessary trade-offs that reduce the quality and resolution of survey data for assessments at large scales. The Peace–Athabasca Delta (PAD) is a large inland wetland complex in northern Alberta, Canada. Despite its geographic isolation, the PAD is threatened by encroachment of oil sands mining in the Athabasca watershed and hydroelectric dams in the Peace watershed. Methods capable of reliably detecting changes in ecosystem health are needed to evaluate and manage risks. Between 2011 and 2016, aquatic macroinvertebrates were sampled across a gradient of wetland flood frequency, applying both microscope-based morphological identification and DNA metabarcoding. By using multispecies occupancy models, we demonstrate that DNA metabarcoding detected a much broader range of taxa and more taxa per sample compared to traditional morphological identification and was essential to identifying significant responses to flood and thermal regimes. We show that family-level occupancy masks high variation among genera and quantify the bias of barcoding primers on the probability of detection in a natural community. Interestingly, patterns of community assembly were nearly random, suggesting a strong role of stochasticity in the dynamics of the metacommunity. This variability seriously compromises effective monitoring at local scales but also reflects resilience to hydrological and thermal variability. Nevertheless, simulations showed the greater efficiency of metabarcoding, particularly at a finer taxonomic resolution, provided the statistical power needed to detect change at the landscape scale.

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          Inference from Iterative Simulation Using Multiple Sequences

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            Replication levels, false presences and the estimation of the presence/absence from eDNA metabarcoding data.

            Environmental DNA (eDNA) metabarcoding is increasingly used to study the present and past biodiversity. eDNA analyses often rely on amplification of very small quantities or degraded DNA. To avoid missing detection of taxa that are actually present (false negatives), multiple extractions and amplifications of the same samples are often performed. However, the level of replication needed for reliable estimates of the presence/absence patterns remains an unaddressed topic. Furthermore, degraded DNA and PCR/sequencing errors might produce false positives. We used simulations and empirical data to evaluate the level of replication required for accurate detection of targeted taxa in different contexts and to assess the performance of methods used to reduce the risk of false detections. Furthermore, we evaluated whether statistical approaches developed to estimate occupancy in the presence of observational errors can successfully estimate true prevalence, detection probability and false-positive rates. Replications reduced the rate of false negatives; the optimal level of replication was strongly dependent on the detection probability of taxa. Occupancy models successfully estimated true prevalence, detection probability and false-positive rates, but their performance increased with the number of replicates. At least eight PCR replicates should be performed if detection probability is not high, such as in ancient DNA studies. Multiple DNA extractions from the same sample yielded consistent results; in some cases, collecting multiple samples from the same locality allowed detecting more species. The optimal level of replication for accurate species detection strongly varies among studies and could be explicitly estimated to improve the reliability of results.
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              Biodiversity soup: metabarcoding of arthropods for rapid biodiversity assessment and biomonitoring

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                Author and article information

                Journal
                Proc Natl Acad Sci U S A
                Proc. Natl. Acad. Sci. U.S.A
                pnas
                pnas
                PNAS
                Proceedings of the National Academy of Sciences of the United States of America
                National Academy of Sciences
                0027-8424
                1091-6490
                14 April 2020
                26 March 2020
                26 March 2020
                : 117
                : 15
                : 8539-8545
                Affiliations
                [1] aEnvironment and Climate Change Canada, Canadian Rivers Institute, Department of Biology, University of New Brunswick , Fredericton, NB E3B 5A3, Canada;
                [2] bLancaster Environment Centre, University of Lancaster , Lancaster LA1 4YW, United Kingdom;
                [3] cEnvironment and Climate Change Canada, Canadian Rivers Institute, Faculty of Forestry and Environmental Management, University of New Brunswick , Fredericton, NB E3B 5A3, Canada;
                [4] dCentre for Environmental Genomics Applications , St. John’s, NL A1A 0R6, Canada;
                [5] eWatershed Hydrology and Ecology Research Division, Environment and Climate Change Canada, University of Victoria , Victoria, BC V8P 5C2, Canada;
                [6] fGreat Lakes Forestry Centre, Natural Resources Canada , Sault Ste. Marie, ON P6A 2E5, Canada;
                [7] gCentre for Biodiversity Genomics, University of Guelph , Guelph, ON N1G 2W1, Canada;
                [8] hDepartment of Integrative Biology, University of Guelph , Guelph, ON N1G 2W1, Canada
                Author notes
                1To whom correspondence may be addressed. Email: alexalbush@ 123456gmail.com .

                Edited by Simon A. Levin, Princeton University, Princeton, NJ, and approved March 2, 2020 (received for review October 29, 2019)

                Author contributions: A.B., M.H., and D.J.B. designed research; A.B. performed research; A.B., W.A.M., Z.G.C., D.L.P., T.M.P., S.S., and M.T.G.W. analyzed data; and A.B., W.A.M., Z.G.C., D.L.P., T.M.P., M.H., and D.J.B. wrote the paper.

                2M.H. and D.J.B. contributed equally to this work.

                Author information
                http://orcid.org/0000-0002-0679-6666
                http://orcid.org/0000-0001-9031-5433
                http://orcid.org/0000-0003-4653-7906
                Article
                201918741
                10.1073/pnas.1918741117
                7165428
                32217735
                7076f457-64cd-4762-bbfe-d4de30379805
                Copyright © 2020 the Author(s). Published by PNAS.

                This open access article is distributed under Creative Commons Attribution License 4.0 (CC BY).

                History
                Page count
                Pages: 7
                Funding
                Funded by: Gouvernement du Canada | Natural Sciences and Engineering Research Council of Canada (NSERC) 501100000038
                Award ID: RGPIN-2019-04891
                Award Recipient : Donald J Baird
                Funded by: Genome Canada (Génome Canada) 100008762
                Award ID: 2419
                Award Recipient : Mehrdad Hajibabaei Award Recipient : Donald J Baird
                Categories
                Biological Sciences
                Ecology

                occupancy,detectability,taxonomic resolution,stochasticity,power analysis

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