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      Using vertebrate environmental DNA from seawater in biomonitoring of marine habitats

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          Abstract

          Conservation and management of marine biodiversity depends on biomonitoring of marine habitats, but current approaches are resource‐intensive and require different approaches for different organisms. Environmental DNA (eDNA) extracted from water samples is an efficient and versatile approach to detecting aquatic animals. In the ocean, eDNA composition reflects local fauna at fine spatial scales, but little is known about the effectiveness of eDNA‐based monitoring of marine communities at larger scales. We investigated the potential of eDNA to characterize and distinguish marine communities at large spatial scales by comparing vertebrate species composition among marine habitats in Qatar, the Arabian Gulf (also known as the Persian Gulf), based on eDNA metabarcoding of seawater samples. We conducted species accumulation analyses to estimate how much of the vertebrate diversity we detected. We obtained eDNA sequences from a diverse assemblage of marine vertebrates, spanning 191 taxa in 73 families. These included rare and endangered species and covered 36% of the bony fish genera previously recorded in the Gulf. Sites of similar habitat type were also similar in eDNA composition. The species accumulation analyses showed that the number of sample replicates was insufficient for some sampling sites but suggested that a few hundred eDNA samples could potentially capture >90% of the marine vertebrate diversity in the study area. Our results confirm that seawater samples contain habitat‐characteristic molecular signatures and that eDNA monitoring can efficiently cover vertebrate diversity at scales relevant to national and regional conservation and management.

          Abstract

          Article impact statement: Environmental DNA provides habitat‐characteristic molecular signatures and can be used efficiently to map marine biodiversity at large spatial scales.

          Translated abstract

          ADN Ambiental de Vertebrados Tomado del Agua Marina para Realizar Biomonitoreos de los Hábitats Marinos

          Resumen

          La conservación y el manejo de la biodiversidad marina depende del biomonitoreo de los hábitats marinos, pero las estrategias actuales requieren de muchos recursos y de diferentes estrategias para diferentes organismos. El ADN ambiental (ADNa) extraído de muestras de agua es una estrategia eficiente y versátil para detectar animales acuáticos. En el océano, la composición del ADNa refleja la fauna local a escalas espaciales finas, pero se sabe poco sobre la efectividad del monitoreo basado en el ADNa de las comunidades marinas a grandes escalas. Investigamos el potencial del ADNa para caracterizar y distinguir las comunidades marinas a escalas espaciales grandes mediante una comparación de la composición de especies de vertebrados entre los hábitats marinos de Qatar, en el Golfo Arábigo (también conocido como el Golfo Persa), con base en el meta‐código de barras del ADNa extraído de muestras de agua de mar. Realizamos análisis de acumulación de especies para estimar cuánta de la diversidad de vertebrados logramos detectar. Obtuvimos secuencias de ADNa de diversos ensamblajes de vertebrados marinos, los cuales abarcaron 191 taxones de 73 familias. Estos taxones incluyeron a especies raras y en peligro de extinción y cubrieron el 36% de los géneros de peces óseos previamente registrados en el golfo. Los sitios con tipos similares de hábitat también fueron similares en cuanto a la composición del ADNa. Los análisis de acumulación de especies mostraron que el número de réplicas de muestras fue insuficiente para algunos sitios de muestreo, pero sugieren que unos cientos de muestras de ADNa podrían capturar potencialmente >90% de la diversidad de vertebrados marinos en el área de estudio. Nuestros resultados confirman que las muestras de agua marina contienen firmas moleculares características del hábitat y que el monitoreo de ADNa puede cubrir eficientemente la diversidad de vertebrados a escalas relevantes para la conservación y el manejo nacional y regional.

          摘要

          海洋生物多样性的保护和管理依赖于对海洋生境的生物监测, 而目前的监测方法都属于资源密集型, 且对不同生物需要采取不同的方法。从水样中提取环境 DNA (eDNA) 是一种调查水生动物高效通用的方法。在海洋中, eDNA 的组成反映了区域精细空间尺度上动物群的情况, 但对于 eDNA 在更大尺度上调查海洋生物群落的有效性尚有待探究。本研究基于 eDNA 条形码技术在海水样品中的应用, 比较了卡塔尔和阿拉伯湾 (又称波斯湾) 海洋生境的脊椎动物组成, 以分析 eDNA 在大空间尺度上描述和分辨海洋生物群落的能力。我们通过物种累积分析估计了我们检测到的脊椎动物多样性。结果显示, 我们获得的 eDNA 序列来自包含 73 个科 191 个类群的海洋脊椎动物群集。这些动物中包括稀有物种和濒危物种, 覆盖了该海湾现有记录中 36% 的硬骨鱼属。生境类型相近的位点检测到的 eDNA 组成也相似。物种累积分析还表明一些采样位点的样品重复个数不足, 但几百个 eDNA 样本可能足够捕获该研究地区 90% 以上的海洋脊椎动物多样性。我们的研究结果证实了海水样本中含有具有生境特点的分子信号, 且 eDNA 检测可以在与国家和区域保护管理所涉及的尺度上有效覆盖脊椎动物多样性。 【翻译: 胡怡思; 审校: 聂永刚】

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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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            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.
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              Scrutinizing key steps for reliable metabarcoding of environmental samples

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                Author and article information

                Contributors
                eva.sigsgaard@bios.au.dk
                pfthomsen@bios.au.dk
                Journal
                Conserv Biol
                Conserv. Biol
                10.1111/(ISSN)1523-1739
                COBI
                Conservation Biology
                John Wiley and Sons Inc. (Hoboken )
                0888-8892
                1523-1739
                24 December 2019
                June 2020
                : 34
                : 3 ( doiID: 10.1111/cobi.v34.3 )
                : 697-710
                Affiliations
                [ 1 ] Natural History Museum of Denmark University of Copenhagen Universitetsparken 15 DK‐2100 Copenhagen Ø Denmark
                [ 2 ]Section for GeoGenetics, Globe Institute, University of Copenhagen, Øster Voldgade 5‐7, DK‐1350 Copenhagen K, Denmark (previously: Centre for GeoGenetics, Natural History Museum of Denmark)
                [ 3 ] School of Ocean Sciences, Bangor University Menai Bridge Anglesey LL59 5AB U.K.
                [ 4 ] Environmental Science Center Qatar University P.O. Box 2713 Doha Qatar
                [ 5 ] Department of Biological and Environmental Sciences Qatar University P.O. Box 2713 Doha Qatar
                [ 6 ] Maersk Oil Research and Technology Centre Al Jazi Tower, Building 20, Zone 60, Street 850, West Bay Doha Qatar
                Author notes
                [*] [* ]Address correspondence to email eva.sigsgaard@ 123456bios.au.dk , pfthomsen@ 123456bios.au.dk
                [†]

                Current address: Department of Bioscience, Aarhus University, Ny Munkegade 116, Building 1540, 8000 Aarhus C, Denmark

                [‡]

                Current address: Rambøll, Hannemanns Allé 53, DK‐2300 Copenhagen S, Denmark

                Author information
                https://orcid.org/0000-0002-1746-7980
                https://orcid.org/0000-0002-3530-013X
                https://orcid.org/0000-0002-9269-2416
                https://orcid.org/0000-0003-2686-5822
                https://orcid.org/0000-0002-0177-0977
                https://orcid.org/0000-0002-9867-4366
                Article
                COBI13437
                10.1111/cobi.13437
                7318234
                31729081
                0e9f68c4-2705-4480-b6b9-a996dddb644c
                © 2019 The Authors. Conservation Biology published by Wiley Periodicals, Inc. on behalf of Society for Conservation Biology.

                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
                : 12 December 2018
                : 12 August 2019
                : 23 September 2019
                Page count
                Figures: 6, Tables: 0, Pages: 14, Words: 7339
                Funding
                Funded by: Maersk Oil , open-funder-registry 10.13039/501100006268;
                Funded by: Qatar National Research Fund , open-funder-registry 10.13039/100008982;
                Award ID: NPRP 7 ‐ 1129 ‐ 1 – 201
                Funded by: Naturvidenskab og Teknologi, Aarhus Universitet , open-funder-registry 10.13039/501100006420;
                Categories
                Contributed Paper
                Contributed Papers
                Custom metadata
                2.0
                June 2020
                Converter:WILEY_ML3GV2_TO_JATSPMC version:5.8.4 mode:remove_FC converted:26.06.2020

                Ecology
                arabian gulf,biomonitoring,fish,metabarcoding,edna,biomonitoreo,golfo arábigo,meta‐código de barras,peces,adna,生物监测,dna 条形码技术,阿拉伯湾,鱼类

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