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      Presence of β-Lactamase-producing Enterobacterales and Salmonella Isolates in Marine Mammals

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          Abstract

          Marine mammals have been described as sentinels of the health of marine ecosystems. Therefore, the aim of this study was to investigate (i) the presence of extended-spectrum β-lactamase (ESBL)- and AmpC-producing Enterobacterales, which comprise several bacterial families important to the healthcare sector, as well as (ii) the presence of Salmonella in these coastal animals. The antimicrobial resistance pheno- and genotypes, as well as biocide susceptibility of Enterobacterales isolated from stranded marine mammals, were determined prior to their rehabilitation. All E. coli isolates ( n = 27) were screened for virulence genes via DNA-based microarray, and twelve selected E. coli isolates were analyzed by whole-genome sequencing. Seventy-one percent of the Enterobacterales isolates exhibited a multidrug-resistant (MDR) pheno- and genotype. The gene bla CMY ( n = 51) was the predominant β-lactamase gene. In addition, bla TEM-1 ( n = 38), bla SHV-33 ( n = 8), bla CTX-M-15 ( n = 7), bla OXA-1 ( n = 7), bla SHV-11 ( n = 3), and bla DHA-1 ( n = 2) were detected. The most prevalent non-β-lactamase genes were sul2 ( n = 38), strA ( n = 34), strB ( n = 34), and tet(A) ( n = 34). Escherichia coli isolates belonging to the pandemic sequence types (STs) ST38, ST167, and ST648 were identified. Among Salmonella isolates ( n = 18), S. Havana was the most prevalent serotype. The present study revealed a high prevalence of MDR bacteria and the presence of pandemic high-risk clones, both of which are indicators of anthropogenic antimicrobial pollution, in marine mammals.

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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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            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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              CARD 2020: antibiotic resistome surveillance with the comprehensive antibiotic resistance database

              Abstract The Comprehensive Antibiotic Resistance Database (CARD; https://card.mcmaster.ca) is a curated resource providing reference DNA and protein sequences, detection models and bioinformatics tools on the molecular basis of bacterial antimicrobial resistance (AMR). CARD focuses on providing high-quality reference data and molecular sequences within a controlled vocabulary, the Antibiotic Resistance Ontology (ARO), designed by the CARD biocuration team to integrate with software development efforts for resistome analysis and prediction, such as CARD’s Resistance Gene Identifier (RGI) software. Since 2017, CARD has expanded through extensive curation of reference sequences, revision of the ontological structure, curation of over 500 new AMR detection models, development of a new classification paradigm and expansion of analytical tools. Most notably, a new Resistomes & Variants module provides analysis and statistical summary of in silico predicted resistance variants from 82 pathogens and over 100 000 genomes. By adding these resistance variants to CARD, we are able to summarize predicted resistance using the information included in CARD, identify trends in AMR mobility and determine previously undescribed and novel resistance variants. Here, we describe updates and recent expansions to CARD and its biocuration process, including new resources for community biocuration of AMR molecular reference data.
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                Author and article information

                Contributors
                Role: Academic Editor
                Journal
                Int J Mol Sci
                Int J Mol Sci
                ijms
                International Journal of Molecular Sciences
                MDPI
                1422-0067
                31 May 2021
                June 2021
                : 22
                : 11
                : 5905
                Affiliations
                [1 ]Institute of Microbiology, University of Veterinary Medicine, 1210 Vienna, Austria; 01245128@ 123456students.vetmeduni.ac.at (O.M.G.); michael.szostak@ 123456vetmeduni.ac.at (M.P.S.); tanja.bernreiter-hofer@ 123456vetmeduni.ac.at (T.B.-H.); koratha@ 123456staff.vetmeduni.ac.at (A.D.J.K.); joachim.spergser@ 123456vetmeduni.ac.at (J.S.)
                [2 ]Marine Mammal Care Center, Los Angeles, CA 90731, USA; lpalmer@ 123456marinemammalcare.org
                [3 ]Austrian Agency for Health and Food Safety (AGES), Institute of Medical Microbiology and Hygiene, 1090 Vienna, Austria; adriana.cabal-rosel@ 123456ages.at (A.C.); werner.ruppitsch@ 123456ages.at (W.R.); franz.allerberger@ 123456ages.at (F.A.)
                [4 ]Austrian Agency for Health and Food Safety (AGES), National Reference Centre for Salmonella, 8010 Graz, Austria; christian.kornschober@ 123456ages.at
                [5 ]Department of Functional Food Products Development, Faculty of Biotechnology and Food Science, Wroclaw University of Environmental and Life Sciences, 51-630 Wroclaw, Poland; maciej.korus@ 123456upwr.edu.pl (M.K.); dusan.misic@ 123456upwr.edu.pl (D.M.)
                [6 ]Department for Farm Animals and Veterinary Public Health, University Clinic for Swine, University of Veterinary Medicine, 1210 Vienna, Austria
                [7 ]Centre for Infection Medicine, Department of Veterinary Medicine, Institute of Microbiology and Epizootics, Freie Universität Berlin, 14163 Berlin, Germany; andrea.fessler@ 123456fu-berlin.de (A.T.F.); stefan.schwarz@ 123456fu-berlin.de (S.S.)
                [8 ]Leibniz Institute of Photonic Technology (IPHT), 07745 Jena, Germany; elke.mueller@ 123456leibniz-ipht.de (E.M.); sascha.braun@ 123456leibniz-ipht.de (S.D.B.); stefan.monecke@ 123456leibniz-ipht.de (S.M.); ralf.ehricht@ 123456leibniz-ipht.de (R.E.)
                [9 ]InfectoGnostics Research Campus, 07743 Jena, Germany
                [10 ]Institute for Medical Microbiology and Hygiene, Technical University of Dresden, 01307 Dresden, Germany
                [11 ]Institute of Physical Chemistry, Friedrich Schiller University Jena, 07743 Jena, Germany
                [12 ]Research Institute of Wildlife Ecology, University of Veterinary Medicine, 1160 Vienna, Austria; cwalzer@ 123456wcs.org
                [13 ]Health Program, Wildlife Conservation Society, Bronx, New York City, NY 10460, USA
                [14 ]College of Veterinary Medicine, Western University of Health Sciences, Pomona, CA 91766-1854, USA; hsmodlaka@ 123456westernu.edu
                Author notes
                [* ]Correspondence: igor.loncaric@ 123456vetmeduni.ac.at ; Tel.: +43-125-077-2115
                Author information
                https://orcid.org/0000-0002-3336-6434
                https://orcid.org/0000-0001-5393-8922
                https://orcid.org/0000-0002-0045-8164
                https://orcid.org/0000-0002-6303-8212
                https://orcid.org/0000-0002-6612-0043
                https://orcid.org/0000-0002-0437-5147
                https://orcid.org/0000-0003-1401-2961
                Article
                ijms-22-05905
                10.3390/ijms22115905
                8199236
                34072783
                035f6421-421d-4a85-96ba-a6d1e06dcefe
                © 2021 by the authors.

                Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license ( https://creativecommons.org/licenses/by/4.0/).

                History
                : 30 April 2021
                : 28 May 2021
                Categories
                Article

                Molecular biology
                antimicrobial resistance,esbl/ampc,wildlife,e. coli,k. pneumoniae
                Molecular biology
                antimicrobial resistance, esbl/ampc, wildlife, e. coli, k. pneumoniae

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