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      Airborne environmental DNA for terrestrial vertebrate community monitoring

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          Summary

          Biodiversity monitoring at the community scale is a critical element of assessing and studying species distributions, ecology, diversity, and movements, and it is key to understanding and tracking environmental and anthropogenic effects on natural ecosystems. 1, 2, 3, 4 Vertebrates in terrestrial ecosystems are experiencing extinctions and declines in both population numbers and sizes due to increasing threats from human activities and environmental change. 5, 6, 7, 8 Terrestrial vertebrate monitoring using existing methods is generally costly and laborious, and although environmental DNA (eDNA) is becoming the tool of choice to assess biodiversity, few sample types effectively capture terrestrial vertebrate diversity. We hypothesized that eDNA captured from air could allow straightforward collection and characterization of terrestrial vertebrate communities. We filtered air at three localities in the Copenhagen Zoo: a stable, outside between the outdoor enclosures, and in the Rainforest House. Through metabarcoding of airborne eDNA, we detected 49 vertebrate species spanning 26 orders and 37 families: 30 mammal, 13 bird, 4 fish, 1 amphibian, and 1 reptile species. These spanned animals kept at the zoo, species occurring in the zoo surroundings, and species used as feed in the zoo. The detected species comprise a range of taxonomic orders and families, sizes, behaviors, and abundances. We found shorter distance to the air sampling device and higher animal biomass to increase the probability of detection. We hereby show that airborne eDNA can offer a fundamentally new way of studying and monitoring terrestrial communities.

          Highlights

          • 49 vertebrate species detected through metabarcoding of airborne eDNA from the zoo

          • Detections included 30 mammal, 13 bird, 4 fish, 1 amphibian, and 1 reptile species

          • 6 to 21 vertebrate species were detected per air filtering sample

          • Shorter geographical distance and higher biomass increased probability of detection

          Abstract

          Lynggaard et al. demonstrate that airborne environmental DNA coupled with metabarcoding and high-throughput sequencing can be used to detect terrestrial vertebrates. The 49 detected species are known to occur in or around the zoo study site. Animals in closer proximity to the sampler and present in larger biomass have higher detection probability.

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          Most cited references51

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          AdapterRemoval v2: rapid adapter trimming, identification, and read merging

          Background As high-throughput sequencing platforms produce longer and longer reads, sequences generated from short inserts, such as those obtained from fossil and degraded material, are increasingly expected to contain adapter sequences. Efficient adapter trimming algorithms are also needed to process the growing amount of data generated per sequencing run. Findings We introduce AdapterRemoval v2, a major revision of AdapterRemoval v1, which introduces (i) striking improvements in throughput, through the use of single instruction, multiple data (SIMD; SSE1 and SSE2) instructions and multi-threading support, (ii) the ability to handle datasets containing reads or read-pairs with different adapters or adapter pairs, (iii) simultaneous demultiplexing and adapter trimming, (iv) the ability to reconstruct adapter sequences from paired-end reads for poorly documented data sets, and (v) native gzip and bzip2 support. Conclusions We show that AdapterRemoval v2 compares favorably with existing tools, while offering superior throughput to most alternatives examined here, both for single and multi-threaded operations. Electronic supplementary material The online version of this article (doi:10.1186/s13104-016-1900-2) contains supplementary material, which is available to authorized users.
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            Double indexing overcomes inaccuracies in multiplex sequencing on the Illumina platform

            Due to the increasing throughput of current DNA sequencing instruments, sample multiplexing is necessary for making economical use of available sequencing capacities. A widely used multiplexing strategy for the Illumina Genome Analyzer utilizes sample-specific indexes, which are embedded in one of the library adapters. However, this and similar multiplex approaches come with a risk of sample misidentification. By introducing indexes into both library adapters (double indexing), we have developed a method that reveals the rate of sample misidentification within current multiplex sequencing experiments. With ~0.3% these rates are orders of magnitude higher than expected and may severely confound applications in cancer genomics and other fields requiring accurate detection of rare variants. We identified the occurrence of mixed clusters on the flow as the predominant source of error. The accuracy of sample identification is further impaired if indexed oligonucleotides are cross-contaminated or if indexed libraries are amplified in bulk. Double-indexing eliminates these problems and increases both the scope and accuracy of multiplex sequencing on the Illumina platform.
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              MEGAN Community Edition - Interactive Exploration and Analysis of Large-Scale Microbiome Sequencing Data

              There is increasing interest in employing shotgun sequencing, rather than amplicon sequencing, to analyze microbiome samples. Typical projects may involve hundreds of samples and billions of sequencing reads. The comparison of such samples against a protein reference database generates billions of alignments and the analysis of such data is computationally challenging. To address this, we have substantially rewritten and extended our widely-used microbiome analysis tool MEGAN so as to facilitate the interactive analysis of the taxonomic and functional content of very large microbiome datasets. Other new features include a functional classifier called InterPro2GO, gene-centric read assembly, principal coordinate analysis of taxonomy and function, and support for metadata. The new program is called MEGAN Community Edition (CE) and is open source. By integrating MEGAN CE with our high-throughput DNA-to-protein alignment tool DIAMOND and by providing a new program MeganServer that allows access to metagenome analysis files hosted on a server, we provide a straightforward, yet powerful and complete pipeline for the analysis of metagenome shotgun sequences. We illustrate how to perform a full-scale computational analysis of a metagenomic sequencing project, involving 12 samples and 800 million reads, in less than three days on a single server. All source code is available here: https://github.com/danielhuson/megan-ce

                Author and article information

                Contributors
                Journal
                Curr Biol
                Curr Biol
                Current Biology
                Cell Press
                0960-9822
                1879-0445
                07 February 2022
                07 February 2022
                : 32
                : 3
                : 701-707.e5
                Affiliations
                [1 ]Section for Evolutionary Genomics, Globe Institute, Faculty of Health and Medical Sciences, University of Copenhagen, 1353 Copenhagen, Denmark
                [2 ]Center for Wild Animal Health, Copenhagen Zoo, 2000 Frederiksberg, Denmark
                [3 ]Department of Chemistry, University of Copenhagen, 2100 Copenhagen, Denmark
                [4 ]Airlabs Denmark, 2200 Copenhagen, Denmark
                [5 ]Section for GeoGenetics, Globe Institute, Faculty of Health and Medical Sciences, University of Copenhagen, 1353 Copenhagen, Denmark
                Author notes
                []Corresponding author christina.lynggaard@ 123456sund.ku.dk
                [∗∗ ]Corresponding author kbohmann@ 123456sund.ku.dk
                [6]

                Twitter: @lynggaardc

                [7]

                Twitter: @kristinebohmann

                [8]

                Lead contact

                Article
                S0960-9822(21)01690-0
                10.1016/j.cub.2021.12.014
                8837273
                34995490
                dceb7a7d-b02a-4e9a-9961-3cb2c9abfdd4
                © 2021 The Author(s)

                This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

                History
                : 7 August 2021
                : 11 October 2021
                : 7 December 2021
                Categories
                Report

                Life sciences
                air samplers,air filtration,amplicon sequencing,bioaerosol,biodiversity,biomonitoring,conservation,edna,high-throughput sequencing,metabarcoding

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