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      Microbial community structure in hadal sediments: high similarity along trench axes and strong changes along redox gradients

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          Abstract

          Hadal trench sediments are hotspots of biogeochemical activity in the deep sea, but the biogeochemical and ecological factors that shape benthic hadal microbial communities remain unknown. Here, we sampled ten hadal sites from two trench regions with a vertical resolution of down to 1 cm. We sequenced 16S rRNA gene amplicons using universal and archaea-specific primer sets and compared the results to biogeochemical parameters. Despite bathymetric and depositional heterogeneity we found a high similarity of microbial communities within each of the two trench axes, while composition at the phylum level varied strongly with sediment depth in conjunction with the redox stratification into oxic, nitrogenous, and ferruginous zones. As a result, communities of a given sediment horizon were more similar to each other across a distance of hundreds of kilometers within each trench, than to those of adjacent horizons from the same sites separated only by centimeters. Total organic carbon content statistically only explained a small part of the variation within and between trenches, and did not explain the community differences observed between the hadal and adjacent shallower sites. Anaerobic taxa increased in abundance at the top of the ferruginous zone, seeded by organisms deposited at the sediment surface and surviving burial through the upper redox zones. While an influence of other potential factors such as geographic isolation, hydrostatic pressure, and non-steady state depositional regimes could not be discerned, redox stratification and diagenesis appear to be the main selective forces that structure community composition in hadal sediments.

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          DADA2: High resolution sample inference from Illumina amplicon data

          We present DADA2, a software package that models and corrects Illumina-sequenced amplicon errors. DADA2 infers sample sequences exactly, without coarse-graining into OTUs, and resolves differences of as little as one nucleotide. In several mock communities DADA2 identified more real variants and output fewer spurious sequences than other methods. We applied DADA2 to vaginal samples from a cohort of pregnant women, revealing a diversity of previously undetected Lactobacillus crispatus variants.
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            Every base matters: assessing small subunit rRNA primers for marine microbiomes with mock communities, time series and global field samples.

            Microbial community analysis via high-throughput sequencing of amplified 16S rRNA genes is an essential microbiology tool. We found the popular primer pair 515F (515F-C) and 806R greatly underestimated (e.g. SAR11) or overestimated (e.g. Gammaproteobacteria) common marine taxa. We evaluated marine samples and mock communities (containing 11 or 27 marine 16S clones), showing alternative primers 515F-Y (5'-GTGYCAGCMGCCGCGGTAA) and 926R (5'-CCGYCAATTYMTTTRAGTTT) yield more accurate estimates of mock community abundances, produce longer amplicons that can differentiate taxa unresolvable with 515F-C/806R, and amplify eukaryotic 18S rRNA. Mock communities amplified with 515F-Y/926R yielded closer observed community composition versus expected (r(2)  = 0.95) compared with 515F-Y/806R (r(2)  ∼ 0.5). Unexpectedly, biases with 515F-Y/806R against SAR11 in field samples (∼4-10-fold) were stronger than in mock communities (∼2-fold). Correcting a mismatch to Thaumarchaea in the 515F-C increased their apparent abundance in field samples, but not as much as using 926R rather than 806R. With plankton samples rich in eukaryotic DNA (> 1 μm size fraction), 18S sequences averaged ∼17% of all sequences. A single mismatch can strongly bias amplification, but even perfectly matched primers can exhibit preferential amplification. We show that beyond in silico predictions, testing with mock communities and field samples is important in primer selection.
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              Simple statistical identification and removal of contaminant sequences in marker-gene and metagenomics data

              Background The accuracy of microbial community surveys based on marker-gene and metagenomic sequencing (MGS) suffers from the presence of contaminants—DNA sequences not truly present in the sample. Contaminants come from various sources, including reagents. Appropriate laboratory practices can reduce contamination, but do not eliminate it. Here we introduce decontam (https://github.com/benjjneb/decontam), an open-source R package that implements a statistical classification procedure that identifies contaminants in MGS data based on two widely reproduced patterns: contaminants appear at higher frequencies in low-concentration samples and are often found in negative controls. Results Decontam classified amplicon sequence variants (ASVs) in a human oral dataset consistently with prior microscopic observations of the microbial taxa inhabiting that environment and previous reports of contaminant taxa. In metagenomics and marker-gene measurements of a dilution series, decontam substantially reduced technical variation arising from different sequencing protocols. The application of decontam to two recently published datasets corroborated and extended their conclusions that little evidence existed for an indigenous placenta microbiome and that some low-frequency taxa seemingly associated with preterm birth were contaminants. Conclusions Decontam improves the quality of metagenomic and marker-gene sequencing by identifying and removing contaminant DNA sequences. Decontam integrates easily with existing MGS workflows and allows researchers to generate more accurate profiles of microbial communities at little to no additional cost. Electronic supplementary material The online version of this article (10.1186/s40168-018-0605-2) contains supplementary material, which is available to authorized users.
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                Author and article information

                Contributors
                schauberger@biology.sdu.dk
                Journal
                ISME J
                ISME J
                The ISME Journal
                Nature Publishing Group UK (London )
                1751-7362
                1751-7370
                8 June 2021
                8 June 2021
                December 2021
                : 15
                : 12
                : 3455-3467
                Affiliations
                [1 ]GRID grid.10825.3e, ISNI 0000 0001 0728 0170, Hadal & Nordcee, Department of Biology, , University of Southern Denmark, ; Odense, Denmark
                [2 ]GRID grid.412785.d, ISNI 0000 0001 0695 6482, Department of Ocean and Environmental Sciences, , Tokyo University of Marine Science and Technology, ; Tokyo, Japan
                [3 ]GRID grid.10825.3e, ISNI 0000 0001 0728 0170, Danish Institute of Advanced Study (DIAS), , University of Southern Denmark, ; Odense, Denmark
                [4 ]GRID grid.10420.37, ISNI 0000 0001 2286 1424, Joint Microbiome Facility of the Medical University of Vienna and the University of Vienna, ; Vienna, Austria
                [5 ]GRID grid.22937.3d, ISNI 0000 0000 9259 8492, Department of Laboratory Medicine, , Medical University of Vienna, ; Vienna, Austria
                [6 ]Univ Brest, CNRS, IFREMER, Microbiology of Extreme Environments Laboratory, Plouzané, France
                [7 ]GRID grid.8390.2, ISNI 0000 0001 2180 5818, Génomique Métabolique, Genoscope, Institut François Jacob, CEA, CNRS, Univ Evry, Université Paris-Saclay, ; Evry, France
                [8 ]GRID grid.503122.7, ISNI 0000 0004 0382 8145, MARBEC, Institut Français de Recherche pour L’Exploitation de la Mer, Univ Montpellier, CNRS, IRD, ; Sète, France
                [9 ]GRID grid.10894.34, ISNI 0000 0001 1033 7684, Alfred Wegener Institute, Helmholtz Center for Polar and Marine Research, ; Bremerhaven, Germany
                [10 ]GRID grid.419529.2, ISNI 0000 0004 0491 3210, Max Planck Institute for Marine Microbiology and Ecology, ; Bremen, Germany
                Author information
                http://orcid.org/0000-0002-2097-4959
                http://orcid.org/0000-0002-7069-893X
                http://orcid.org/0000-0002-0846-1202
                http://orcid.org/0000-0003-4307-4782
                http://orcid.org/0000-0002-5571-5228
                http://orcid.org/0000-0002-8744-3116
                http://orcid.org/0000-0001-7562-3454
                http://orcid.org/0000-0001-5814-8452
                http://orcid.org/0000-0002-4621-0586
                http://orcid.org/0000-0002-1221-7077
                Article
                1021
                10.1038/s41396-021-01021-w
                8629969
                34103697
                e5398d05-ca41-4481-8972-867cba1d4a6f
                © The Author(s) 2021

                Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

                History
                : 17 December 2020
                : 11 May 2021
                : 20 May 2021
                Funding
                Funded by: FundRef https://doi.org/10.13039/100010663, EC | EU Framework Programme for Research and Innovation H2020 | H2020 Priority Excellent Science | H2020 European Research Council (H2020 Excellent Science - European Research Council);
                Award ID: #669947
                Award ID: #669947
                Award Recipient :
                Funded by: FundRef https://doi.org/10.13039/501100001732, Danmarks Grundforskningsfond (Danish National Research Foundation);
                Award ID: DNRF145
                Award ID: DNRF145
                Award ID: DNRF145
                Award Recipient :
                Funded by: Missing funding sources can be found in the acknowledgements: "Furthermore, our gratitude goes to the captains, crews and scientific personnel of the RV Sonne (SO261; ship time provided by BMBF, Germany, awarded to Frank Wenzhoefer, Mathias Zabel and Ronnie N. Glud) and RV Tangaroa (TAN1711; shiptime partly funded by Coasts & Oceans Centre of New Zealand’s National Institute of Water & Atmospheric Research (NIWA), awarded to Ashley A. Rowden and Ronnie N. Glud). Financial support for the study was provided by HADES-ERC Advanced grant “Benthic diagenesis and microbiology of hadal trenches” #669947 and the Danish National Research Foundation grant DNRF145 to establish the Danish Center for Hadal Research, both awarded to Ronnie N. Glud. This work is part of the “Pourquoi Pas les Abysses?” project funded by Ifremer, and the project eDNAbyss (AP2016 -228) funded by France Génomique (ANR-10-INBS-09) and Genoscope-CEA."
                Funded by: FundRef https://doi.org/10.13039/100011199, EC | EC Seventh Framework Programm | FP7 Ideas: European Research Council (FP7-IDEAS-ERC - Specific Programme: "Ideas" Implementing the Seventh Framework Programme of the European Community for Research, Technological Development and Demonstration Activities (2007 to 2013));
                Award ID: #669947
                Award Recipient :
                Funded by: Please see acknowlegements: "Furthermore, our gratitude goes to the captains, crews and scientific personnel of the RV Sonne (SO261; ship time provided by BMBF, Germany, awarded to Frank Wenzhoefer, Mathias Zabel and Ronnie N. Glud) and RV Tangaroa (TAN1711; shiptime partly funded by Coasts & Oceans Centre of New Zealand’s National Institute of Water & Atmospheric Research (NIWA), awarded to Ashley A. Rowden and Ronnie N. Glud). Financial support for the study was provided by HADES-ERC Advanced grant “Benthic diagenesis and microbiology of hadal trenches” #669947 and the Danish National Research Foundation grant DNRF145 to establish the Danish Center for Hadal Research, both awarded to Ronnie N. Glud. This work is part of the “Pourquoi Pas les Abysses?” project funded by Ifremer, and the project eDNAbyss (AP2016 -228) funded by France Génomique (ANR-10-INBS-09) and Genoscope-CEA."
                Funded by: As described in the acknowledgement section: This work is part of the “Pourquoi Pas les Abysses?” project funded by Ifremer, and the project eDNAbyss (AP2016 -228) funded by France Génomique (ANR-10-INBS-09) and Genoscope-CEA.
                Funded by: Please again see in the acknowledgements: "Furthermore, our gratitude goes to the captains, crews and scientific personnel of the RV Sonne (SO261; ship time provided by BMBF, Germany, awarded to Frank Wenzhoefer, Mathias Zabel and Ronnie N. Glud) and RV Tangaroa (TAN1711; shiptime partly funded by Coasts & Oceans Centre of New Zealand’s National Institute of Water & Atmospheric Research (NIWA), awarded to Ashley A. Rowden and Ronnie N. Glud)."
                Funded by: Please again see in acknowledgements: Furthermore, our gratitude goes to the captains, crews and scientific personnel of the RV Sonne (SO261; ship time provided by BMBF, Germany, awarded to Frank Wenzhoefer, Mathias Zabel and Ronnie N. Glud) and RV Tangaroa (TAN1711; shiptime partly funded by Coasts & Oceans Centre of New Zealand’s National Institute of Water & Atmospheric Research (NIWA), awarded to Ashley A. Rowden and Ronnie N. Glud). Financial support for the study was provided by HADES-ERC Advanced grant “Benthic diagenesis and microbiology of hadal trenches” #669947 and the Danish National Research Foundation grant DNRF145 to establish the Danish Center for Hadal Research, both awarded to Ronnie N. Glud. This work is part of the “Pourquoi Pas les Abysses?” project funded by Ifremer, and the project eDNAbyss (AP2016 -228) funded by France Génomique (ANR-10-INBS-09) and Genoscope-CEA.
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                © The Author(s), under exclusive licence to International Society for Microbial Ecology 2021

                Microbiology & Virology
                water microbiology,microbial ecology,microbiome,biogeochemistry
                Microbiology & Virology
                water microbiology, microbial ecology, microbiome, biogeochemistry

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