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      Metagenomic tracking of antibiotic resistance genes through a pre‐harvest vegetable production system: an integrated lab‐, microcosm‐ and greenhouse‐scale analysis

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          Summary

          Prior research demonstrated the potential for agricultural production systems to contribute to the environmental spread of antibiotic resistance genes (ARGs). However, there is a need for integrated assessment of critical management points for minimizing this potential. Shotgun metagenomic sequencing data were analysed to comprehensively compare total ARG profiles characteristic of amendments (manure or compost) derived from either beef or dairy cattle (with and without dosing antibiotics according to conventional practice), soil (loamy sand or silty clay loam) and vegetable (lettuce or radish) samples collected across studies carried out at laboratory‐, microcosm‐ and greenhouse‐scale. Vegetables carried the greatest diversity of ARGs ( n = 838) as well as the most ARG‐mobile genetic element co‐occurrences ( n = 945). Radishes grown in manure‐ or compost‐amended soils harboured a higher relative abundance of total (0.91 and 0.91 ARGs/16S rRNA gene) and clinically relevant ARGs than vegetables from other experimental conditions (average: 0.36 ARGs/16S rRNA gene). Lettuce carried the highest relative abundance of pathogen gene markers among the metagenomes examined. Total ARG relative abundances were highest on vegetables grown in loamy sand receiving antibiotic‐treated beef amendments. The findings emphasize that additional barriers, such as post‐harvest processes, merit further study to minimize potential exposure to consumers.

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          Trimmomatic: a flexible trimmer for Illumina sequence data

          Motivation: Although many next-generation sequencing (NGS) read preprocessing tools already existed, we could not find any tool or combination of tools that met our requirements in terms of flexibility, correct handling of paired-end data and high performance. We have developed Trimmomatic as a more flexible and efficient preprocessing tool, which could correctly handle paired-end data. Results: The value of NGS read preprocessing is demonstrated for both reference-based and reference-free tasks. Trimmomatic is shown to produce output that is at least competitive with, and in many cases superior to, that produced by other tools, in all scenarios tested. Availability and implementation: Trimmomatic is licensed under GPL V3. It is cross-platform (Java 1.5+ required) and available at http://www.usadellab.org/cms/index.php?page=trimmomatic Contact: usadel@bio1.rwth-aachen.de Supplementary information: Supplementary data are available at Bioinformatics online.
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            Fast gapped-read alignment with Bowtie 2.

            As the rate of sequencing increases, greater throughput is demanded from read aligners. The full-text minute index is often used to make alignment very fast and memory-efficient, but the approach is ill-suited to finding longer, gapped alignments. Bowtie 2 combines the strengths of the full-text minute index with the flexibility and speed of hardware-accelerated dynamic programming algorithms to achieve a combination of high speed, sensitivity and accuracy.
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              Fast and sensitive protein alignment using DIAMOND.

              The alignment of sequencing reads against a protein reference database is a major computational bottleneck in metagenomics and data-intensive evolutionary projects. Although recent tools offer improved performance over the gold standard BLASTX, they exhibit only a modest speedup or low sensitivity. We introduce DIAMOND, an open-source algorithm based on double indexing that is 20,000 times faster than BLASTX on short reads and has a similar degree of sensitivity.
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                Author and article information

                Contributors
                apruden@vt.edu
                Journal
                Environ Microbiol
                Environ Microbiol
                10.1111/(ISSN)1462-2920
                EMI
                Environmental Microbiology
                John Wiley & Sons, Inc. (Hoboken, USA )
                1462-2912
                1462-2920
                18 May 2022
                August 2022
                : 24
                : 8 , Special Issue on Microbial Ecology of Plants ( doiID: 10.1111/emi.v24.8 )
                : 3705-3721
                Affiliations
                [ 1 ] Department of Civil and Environmental Engineering Virginia Tech Blacksburg VA USA
                [ 2 ] Department of Biological Systems Engineering Virginia Tech Blacksburg VA USA
                [ 3 ] Department of Animal Sciences, School of Agriculture, Policy and Development University of Reading Reading RG6 6AR UK
                [ 4 ] Department of Food Science and Technology Virginia Tech Blacksburg VA USA
                [ 5 ] Department of Crop and Soil Environmental Sciences Virginia Tech Blacksburg VA USA
                [ 6 ] Department of Dairy Science Virginia Tech Blacksburg VA USA
                Author notes
                [*] [* ] For correspondence. E‐mail apruden@ 123456vt.edu ; Tel. (540)231‐3980; Fax (540)231‐3980.

                Author information
                https://orcid.org/0000-0002-5441-4634
                https://orcid.org/0000-0002-3191-6244
                Article
                EMI16022
                10.1111/1462-2920.16022
                9541739
                35466491
                f0738936-4fcb-42ee-a358-b43d19361c89
                © 2022 The Authors. Environmental Microbiology published by Society for Applied Microbiology and 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
                : 14 April 2022
                : 21 July 2021
                : 18 April 2022
                Page count
                Figures: 5, Tables: 0, Pages: 17, Words: 12078
                Funding
                Funded by: National Institute of Food and Agriculture , doi 10.13039/100005825;
                Award ID: 2015‐68003‐23050
                Award ID: 2017‐68003‐26498
                Funded by: Spring Point Partners, LLC Water Innovators Program
                Funded by: Virginia Tech ICTAS Centre
                Categories
                Research Article
                Research Articles
                Custom metadata
                2.0
                August 2022
                Converter:WILEY_ML3GV2_TO_JATSPMC version:6.2.0 mode:remove_FC converted:07.10.2022

                Microbiology & Virology
                Microbiology & Virology

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