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      Assessing eukaryotic biodiversity in the Florida Keys National Marine Sanctuary through environmental DNA metabarcoding

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          Abstract

          Environmental DNA (eDNA) is the DNA suspended in the environment (e.g., water column), which includes cells, gametes, and other material derived from but not limited to shedding of tissue, scales, mucus, and fecal matter. Amplifying and sequencing marker genes (i.e., metabarcoding) from eDNA can reveal the wide range of taxa present in an ecosystem through analysis of a single water sample. Metabarcoding of eDNA provides higher resolution data than visual surveys, aiding in assessments of ecosystem health. This study conducted eDNA metabarcoding of two molecular markers (cytochrome c oxidase I (COI) and 18S ribosomal RNA (rRNA) genes) to survey eukaryotic diversity across multiple trophic levels in surface water samples collected at three sites along the coral reef tract within the Florida Keys National Marine Sanctuary (FKNMS) during four research cruises in 2015. The 18S rRNA gene sequences recovered 785 genera while the COI gene sequences recovered 115 genera, with only 33 genera shared between the two datasets, emphasizing the complementarity of these marker genes. Community composition for both genetic markers clustered by month of sample collection, suggesting that temporal variation has a larger effect on biodiversity than spatial variability in the FKNMS surface waters. Sequences from both marker genes were dominated by copepods, but each marker recovered distinct phytoplankton groups, with 18S rRNA gene sequences dominated by dinoflagellates and COI sequences dominated by coccolithophores. Although eDNA samples were collected from surface waters, many benthic species such as sponges, crustaceans, and corals were identified. These results show the utility of eDNA metabarcoding for cataloging biodiversity to establish an ecosystem baseline against which future samples can be compared in order to monitor community changes.

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          Most cited references31

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          A molecular view of microbial diversity and the biosphere.

          N Pace (1997)
          Over three decades of molecular-phylogenetic studies, researchers have compiled an increasingly robust map of evolutionary diversification showing that the main diversity of life is microbial, distributed among three primary relatedness groups or domains: Archaea, Bacteria, and Eucarya. The general properties of representatives of the three domains indicate that the earliest life was based on inorganic nutrition and that photosynthesis and use of organic compounds for carbon and energy metabolism came comparatively later. The application of molecular-phylogenetic methods to study natural microbial ecosystems without the traditional requirement for cultivation has resulted in the discovery of many unexpected evolutionary lineages; members of some of these lineages are only distantly related to known organisms but are sufficiently abundant that they are likely to have impact on the chemistry of the biosphere.
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            Detection of a Diverse Marine Fish Fauna Using Environmental DNA from Seawater Samples

            Marine ecosystems worldwide are under threat with many fish species and populations suffering from human over-exploitation. This is greatly impacting global biodiversity, economy and human health. Intriguingly, marine fish are largely surveyed using selective and invasive methods, which are mostly limited to commercial species, and restricted to particular areas with favourable conditions. Furthermore, misidentification of species represents a major problem. Here, we investigate the potential of using metabarcoding of environmental DNA (eDNA) obtained directly from seawater samples to account for marine fish biodiversity. This eDNA approach has recently been used successfully in freshwater environments, but never in marine settings. We isolate eDNA from ½-litre seawater samples collected in a temperate marine ecosystem in Denmark. Using next-generation DNA sequencing of PCR amplicons, we obtain eDNA from 15 different fish species, including both important consumption species, as well as species rarely or never recorded by conventional monitoring. We also detect eDNA from a rare vagrant species in the area; European pilchard (Sardina pilchardus). Additionally, we detect four bird species. Records in national databases confirmed the occurrence of all detected species. To investigate the efficiency of the eDNA approach, we compared its performance with 9 methods conventionally used in marine fish surveys. Promisingly, eDNA covered the fish diversity better than or equal to any of the applied conventional methods. Our study demonstrates that even small samples of seawater contain eDNA from a wide range of local fish species. Finally, in order to examine the potential dispersal of eDNA in oceans, we performed an experiment addressing eDNA degradation in seawater, which shows that even small (100-bp) eDNA fragments degrades beyond detectability within days. Although further studies are needed to validate the eDNA approach in varying environmental conditions, our findings provide a strong proof-of-concept with great perspectives for future monitoring of marine biodiversity and resources.
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              Swarm: robust and fast clustering method for amplicon-based studies

              Popular de novo amplicon clustering methods suffer from two fundamental flaws: arbitrary global clustering thresholds, and input-order dependency induced by centroid selection. Swarm was developed to address these issues by first clustering nearly identical amplicons iteratively using a local threshold, and then by using clusters’ internal structure and amplicon abundances to refine its results. This fast, scalable, and input-order independent approach reduces the influence of clustering parameters and produces robust operational taxonomic units.
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                Author and article information

                Contributors
                anni.djurhuus@gmail.com
                Journal
                Ecol Evol
                Ecol Evol
                10.1002/(ISSN)2045-7758
                ECE3
                Ecology and Evolution
                John Wiley and Sons Inc. (Hoboken )
                2045-7758
                15 January 2019
                February 2019
                : 9
                : 3 ( doiID: 10.1002/ece3.2019.9.issue-3 )
                : 1029-1040
                Affiliations
                [ 1 ] College of Marine Science University of South Florida Saint Petersburg Florida
                [ 2 ] Stanford Center for Ocean Solutions Stanford University Stanford California
                [ 3 ] Department of Civil and Environmental Engineering Stanford University Stanford California
                [ 4 ] Florida Fish and Wildlife Conservation Commission‐Fish and Wildlife Research Institute Saint Petersburg Florida
                [ 5 ] Rosenstiel School of Marine and Atmospheric Science University of Miami Miami Florida
                [ 6 ] NOAA Atlantic Oceanographic and Meteorological Laboratory Miami Florida
                Author notes
                [*] [* ] Correspondence

                Anni Djurhuus, University of South Florida, College of Marine Science, Saint Petersburg, FL.

                Email: anni.djurhuus@ 123456gmail.com

                Author information
                http://orcid.org/0000-0001-6841-2659
                http://orcid.org/0000-0002-3517-7522
                http://orcid.org/0000-0003-3210-2899
                Article
                ECE34742
                10.1002/ece3.4742
                6374654
                30805138
                fa6e09e3-68b6-4b5f-8f98-c0e3d9ac0899
                © 2019 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
                : 18 April 2018
                : 02 October 2018
                : 10 October 2018
                Page count
                Figures: 5, Tables: 2, Pages: 12, Words: 8290
                Funding
                Funded by: Bureau of Ocean Energy Management
                Funded by: National Ocean Partnership Program
                Award ID: 2014‐2003803
                Funded by: Integrated Ocean Observing System
                Funded by: National Aeronautics and Space Administration
                Award ID: NNX14AP62A
                Funded by: National Oceanic and Atmospheric Administration
                Categories
                Original Research
                Original Research
                Custom metadata
                2.0
                ece34742
                February 2019
                Converter:WILEY_ML3GV2_TO_NLMPMC version:5.5.8 mode:remove_FC converted:14.02.2019

                Evolutionary Biology
                edna,18s rrna gene,cytochrome c oxidase i,monitoring
                Evolutionary Biology
                edna, 18s rrna gene, cytochrome c oxidase i, monitoring

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