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      Surveillance of Enterococcus spp. reveals distinct species and antimicrobial resistance diversity across a One-Health continuum

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          Abstract

          For a One-Health investigation of antimicrobial resistance (AMR) in Enterococcus spp., isolates from humans and beef cattle along with abattoirs, manured fields, natural streams, and wastewater from both urban and cattle feedlot sources were collected over two years. Species identification of Enterococcus revealed distinct associations across the continuum. Of the 8430 isolates collected, Enterococcus faecium and Enterococcus faecalis were the main species in urban wastewater (90%) and clinical human isolates (99%); Enterococcus hirae predominated in cattle (92%) and feedlot catch-basins (60%), whereas natural streams harbored environmental Enterococcus spp. Whole-genome sequencing of E. faecalis (n = 366 isolates) and E. faecium (n = 342 isolates), revealed source clustering of isolates, indicative of distinct adaptation to their respective environments. Phenotypic resistance to tetracyclines and macrolides encoded by tet(M) and erm(B) respectively, was prevalent among Enterococcus spp. regardless of source. For E. faecium from cattle, resistance to β-lactams and quinolones was observed among 3% and 8% of isolates respectively, compared to 76% and 70% of human clinical isolates. Clinical vancomycin-resistant E. faecium exhibited high rates of multi-drug resistance, with resistance to all β-lactam, macrolides, and quinolones tested. Differences in the AMR profiles among isolates reflected antimicrobial use practices in each sector of the One-Health continuum.

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          SPAdes: a new genome assembly algorithm and its applications to single-cell sequencing.

          The lion's share of bacteria in various environments cannot be cloned in the laboratory and thus cannot be sequenced using existing technologies. A major goal of single-cell genomics is to complement gene-centric metagenomic data with whole-genome assemblies of uncultivated organisms. Assembly of single-cell data is challenging because of highly non-uniform read coverage as well as elevated levels of sequencing errors and chimeric reads. We describe SPAdes, a new assembler for both single-cell and standard (multicell) assembly, and demonstrate that it improves on the recently released E+V-SC assembler (specialized for single-cell data) and on popular assemblers Velvet and SoapDeNovo (for multicell data). SPAdes generates single-cell assemblies, providing information about genomes of uncultivatable bacteria that vastly exceeds what may be obtained via traditional metagenomics studies. SPAdes is available online ( http://bioinf.spbau.ru/spades ). It is distributed as open source software.
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            Prokka: rapid prokaryotic genome annotation.

            T Seemann (2014)
            The multiplex capability and high yield of current day DNA-sequencing instruments has made bacterial whole genome sequencing a routine affair. The subsequent de novo assembly of reads into contigs has been well addressed. The final step of annotating all relevant genomic features on those contigs can be achieved slowly using existing web- and email-based systems, but these are not applicable for sensitive data or integrating into computational pipelines. Here we introduce Prokka, a command line software tool to fully annotate a draft bacterial genome in about 10 min on a typical desktop computer. It produces standards-compliant output files for further analysis or viewing in genome browsers. Prokka is implemented in Perl and is freely available under an open source GPLv2 license from http://vicbioinformatics.com/. © The Author 2014. Published by Oxford University Press. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com.
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              In silico detection and typing of plasmids using PlasmidFinder and plasmid multilocus sequence typing.

              In the work presented here, we designed and developed two easy-to-use Web tools for in silico detection and characterization of whole-genome sequence (WGS) and whole-plasmid sequence data from members of the family Enterobacteriaceae. These tools will facilitate bacterial typing based on draft genomes of multidrug-resistant Enterobacteriaceae species by the rapid detection of known plasmid types. Replicon sequences from 559 fully sequenced plasmids associated with the family Enterobacteriaceae in the NCBI nucleotide database were collected to build a consensus database for integration into a Web tool called PlasmidFinder that can be used for replicon sequence analysis of raw, contig group, or completely assembled and closed plasmid sequencing data. The PlasmidFinder database currently consists of 116 replicon sequences that match with at least at 80% nucleotide identity all replicon sequences identified in the 559 fully sequenced plasmids. For plasmid multilocus sequence typing (pMLST) analysis, a database that is updated weekly was generated from www.pubmlst.org and integrated into a Web tool called pMLST. Both databases were evaluated using draft genomes from a collection of Salmonella enterica serovar Typhimurium isolates. PlasmidFinder identified a total of 103 replicons and between zero and five different plasmid replicons within each of 49 S. Typhimurium draft genomes tested. The pMLST Web tool was able to subtype genomic sequencing data of plasmids, revealing both known plasmid sequence types (STs) and new alleles and ST variants. In conclusion, testing of the two Web tools using both fully assembled plasmid sequences and WGS-generated draft genomes showed them to be able to detect a broad variety of plasmids that are often associated with antimicrobial resistance in clinically relevant bacterial pathogens. Copyright © 2014, American Society for Microbiology. All Rights Reserved.
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                Author and article information

                Contributors
                tim.mcallister@canada.ca
                Journal
                Sci Rep
                Sci Rep
                Scientific Reports
                Nature Publishing Group UK (London )
                2045-2322
                3 March 2020
                3 March 2020
                2020
                : 10
                : 3937
                Affiliations
                [1 ]ISNI 0000 0001 1302 4958, GRID grid.55614.33, Lethbridge Research and Development Centre, , Agriculture and Agri-Food Canada, ; 5403 1st Avenue South, Lethbridge, AB T1J 4P4 Canada
                [2 ]Alberta Agriculture and Forestry, 100, 5401 1st Avenue South, Lethbridge, AB T1J 4V6 Canada
                [3 ]Canadian Food Inspection Agency, National Center for Animal Disease, Lethbridge Laboratory, Township Rd 9-1, Lethbridge, AB T1J3Z4 Canada
                [4 ]ISNI 0000 0001 0805 4386, GRID grid.415368.d, National Microbiology Laboratory, , Public Health Agency of Canada, ; 1015 Arlington Street, Winnipeg, MB R3E 3R2 Canada
                [5 ]Feedlot Health Management Services, Okotoks, AB Canada
                [6 ]ISNI 0000 0001 1302 4958, GRID grid.55614.33, Lacombe Research and Development Centre, , Agriculture and Agri-Food Canada, ; 6000C and E Trail, Lacombe, AB T4L 1W1 Canada
                [7 ]ISNI 0000 0004 1936 7697, GRID grid.22072.35, Cumming School of Medicine, , University of Calgary, ; 3280 Hospital Drive NW, Calgary, Alberta Canada
                [8 ]ISNI 0000 0001 0693 8815, GRID grid.413574.0, Calgary Laboratory Services (CLS), , Alberta Health Services, ; 3535 Research Rd NW, Calgary, AB T2L 2K8 Canada
                Article
                61002
                10.1038/s41598-020-61002-5
                7054549
                32127598
                48dcafc6-332d-4f02-b321-2ce01704a82b
                © The Author(s) 2020

                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
                : 19 October 2019
                : 13 February 2020
                Funding
                Funded by: FundRef http://dx.doi.org/10.13039/501100005019, Beef Cattle Research Council;
                Award ID: Project FOS 10.13
                Award ID: Project FOS 10.13
                Award ID: Project FOS 10.13
                Award ID: Project FOS 10.13
                Award ID: Project FOS 10.13
                Award ID: Project FOS 10.13
                Award ID: Project FOS 10.13
                Award ID: Project FOS 10.13
                Award ID: Project FOS 10.13
                Award ID: Project FOS 10.13
                Award ID: Project FOS 10.13
                Award Recipient :
                Funded by: Genomics Research and Development Initiative of the Government of Canada
                Award ID: 001
                Award ID: 001
                Award ID: 001
                Award ID: 001
                Award ID: 001
                Award ID: 001
                Award ID: 001
                Award Recipient :
                Categories
                Article
                Custom metadata
                © The Author(s) 2020

                Uncategorized
                microbiology,antimicrobials,antimicrobial resistance
                Uncategorized
                microbiology, antimicrobials, antimicrobial resistance

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