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      MALDI-TOF MS and genomic analysis can make the difference in the clarification of canine brucellosis outbreaks

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          Abstract

          Brucellosis is one of the most common bacterial zoonoses worldwide affecting not only livestock and wildlife but also pets. Canine brucellosis is characterized by reproductive failure in dogs. Human Brucella canis infections are rarely reported but probably underestimated due to insufficient diagnostic surveillance. To improve diagnostics, we investigated dogs in a breeding kennel that showed clinical manifestations of brucellosis and revealed positive blood cultures. As an alternative to the time-consuming and hazardous classical identification procedures, a newly developed species-specific intact-cell matrix-assisted laser desorption/ionization–time of flight mass spectrometry analysis was applied, which allowed for rapid identification of B. canis and differentiation from closely related B. suis biovar 1. High-throughput sequencing and comparative genomics using single nucleotide polymorphism analysis clustered our isolates together with canine and human strains from various Central and South American countries in a distinct sub-lineage. Hence, molecular epidemiology clearly defined the outbreak cluster and demonstrated the endemic situation in South America. Our study illustrates that MALDI-TOF MS analysis using a validated in-house reference database facilitates rapid B. canis identification at species level. Additional whole genome sequencing provides more detailed outbreak information and leads to a deeper understanding of the epidemiology of canine brucellosis.

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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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            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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                Author and article information

                Contributors
                Dirk.Hofreuter@bfr.bund.de
                Journal
                Sci Rep
                Sci Rep
                Scientific Reports
                Nature Publishing Group UK (London )
                2045-2322
                6 November 2020
                6 November 2020
                2020
                : 10
                : 19246
                Affiliations
                [1 ]GRID grid.417830.9, ISNI 0000 0000 8852 3623, Department of Biological Safety, , German Federal Institute for Risk Assessment, ; Berlin, Germany
                [2 ]GRID grid.11899.38, ISNI 0000 0004 1937 0722, Department of Preventive Veterinary Medicine and Animal Health, Faculty of Veterinary Medicine and Animal Science, , University of São Paulo, ; São Paulo, Brazil
                [3 ]GRID grid.11899.38, ISNI 0000 0004 1937 0722, Department of Veterinary Medicine, Faculty of Animal Science and Food Engineering, , University of São Paulo, ; Pirassununga, Brazil
                [4 ]GRID grid.412301.5, ISNI 0000 0000 8653 1507, Department of Internal Medicine, , RWTH Aachen University Hospital, ; Aachen, Germany
                Article
                75960
                10.1038/s41598-020-75960-3
                7648634
                33159111
                4dad27f0-fbb2-4c5a-a92f-10ee3c99528b
                © 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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/.

                History
                : 6 June 2020
                : 13 October 2020
                Funding
                Funded by: Brazilian National Council for Scientific and Technological Development (CNPq)
                Funded by: German Federal Ministry of Education and Research (BMBF)
                Award ID: FKZ 13N13982
                Funded by: São Paulo Research Foundation (FAPESP)
                Award ID: 2016/01276-8, 2016/13804-9
                Funded by: Projekt DEAL
                Categories
                Article
                Custom metadata
                © The Author(s) 2020

                Uncategorized
                genomic analysis,mass spectrometry,sequencing,microbiology techniques,bacteria,bacteriology,infectious-disease diagnostics,pathogens,diagnostic markers,microbiology,bacterial infection,diseases,infectious diseases,pathogenesis,infection

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