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      Whole-genome sequencing of Leptospira interrogans from southern Brazil: genetic features of a highly virulent strain

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          Abstract

          BACKGROUND

          Leptospirosis is the most widespread zoonotic disease. It is caused by infection with pathogenic Leptospira species, of which over 300 serovars have been described. The accurate identification of the causative Leptospira spp. is required to ascertain the pathogenic status of the local isolates.

          OBJECTIVES

          This study aimed to obtain the complete genome sequence of a virulent Leptospira interrogans strain isolated from southern Brazil and to describe its genetic features.

          METHODS

          The whole genome was sequenced by next-generation sequencing (Ion Torrent). The genome was assembled, scaffolded, annotated, and manually reviewed. Mutations were identified based on a variant calling analysis using the genome of L. interrogans strain Fiocruz L1-130 as a reference.

          FINDINGS

          The entire genome had an average GC content of 35%. The variant calling analysis identified 119 single nucleotide polymorphisms (SNPs), from which 30 led to a missense mutation. The structural analyses identified potential evidence of genomic inversions, translocations, and deletions in both the chromosomes.

          MAIN CONCLUSIONS

          The genome properties provide comprehensive information about the local isolates of Leptospira spp., and thereby, could facilitate the identification of new targets for the development of diagnostic kits and vaccines.

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          Most cited references28

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          Artemis: sequence visualization and annotation

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            Leptospira and leptospirosis.

            Leptospirosis is the most wide spread zoonosis worldwide; it is present in all continents except Antarctica and evidence for the carriage of Leptospira has been found in virtually all mammalian species examined. Humans most commonly become infected through occupational, recreational, or domestic contact with the urine of carrier animals, either directly or via contaminated water or soil. Leptospires are thin, helical bacteria classified into at least 12 pathogenic and 4 saprophytic species, with more than 250 pathogenic serovars. Immunity following infection is generally, but not exclusively, mediated by antibody against leptospiral LPS and restricted to antigenically related serovars. Vaccines currently available consist of killed whole cell bacterins which are used widely in animals, but less so in humans. Current work with recombinant protein antigens shows promise for the development of vaccines based on defined protective antigens. The cellular and molecular basis for virulence remains poorly understood, but comparative genomics of pathogenic and saprophytic species suggests that Leptospira expresses unique virulence determinants. However, the recent development of defined mutagenesis systems for Leptospira heralds the potential for gaining a much improved understanding of pathogenesis in leptospirosis. Copyright 2009 Elsevier B.V. All rights reserved.
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              Comparative genomics of two Leptospira interrogans serovars reveals novel insights into physiology and pathogenesis.

              Leptospira species colonize a significant proportion of rodent populations worldwide and produce life-threatening infections in accidental hosts, including humans. Complete genome sequencing of Leptospira interrogans serovar Copenhageni and comparative analysis with the available Leptospira interrogans serovar Lai genome reveal that despite overall genetic similarity there are significant structural differences, including a large chromosomal inversion and extensive variation in the number and distribution of insertion sequence elements. Genome sequence analysis elucidates many of the novel aspects of leptospiral physiology relating to energy metabolism, oxygen tolerance, two-component signal transduction systems, and mechanisms of pathogenesis. A broad array of transcriptional regulation proteins and two new families of afimbrial adhesins which contribute to host tissue colonization in the early steps of infection were identified. Differences in genes involved in the biosynthesis of lipopolysaccharide O side chains between the Copenhageni and Lai serovars were identified, offering an important starting point for the elucidation of the organism's complex polysaccharide surface antigens. Differences in adhesins and in lipopolysaccharide might be associated with the adaptation of serovars Copenhageni and Lai to different animal hosts. Hundreds of genes encoding surface-exposed lipoproteins and transmembrane outer membrane proteins were identified as candidates for development of vaccines for the prevention of leptospirosis.
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                Author and article information

                Journal
                Mem Inst Oswaldo Cruz
                Mem. Inst. Oswaldo Cruz
                mioc
                Memórias do Instituto Oswaldo Cruz
                Instituto Oswaldo Cruz, Ministério da Saúde
                0074-0276
                1678-8060
                February 2018
                February 2018
                : 113
                : 2
                : 80-86
                Affiliations
                [1]Universidade Federal de Pelotas, Centro de Desenvolvimento Tecnológico, Núcleo de Biotecnologia, Pelotas, RS, Brasil
                Author notes
                [+ ] Corresponding author: sergiojorgevet@ 123456hotmail.com

                AUTHORS’ CONTRIBUTION

                SJ, FSK, NRO, VFC, LSP and OAD designed the study and wrote the manuscript; SJ and NRO performed the sequencing experiment; FSK, GOSVN, AMG, CDS and RDSW performed the bioinformatics analysis and created the figures and tables; KFR isolated the Piscina strain. All authors contributed to and revised the manuscript.

                The authors certify that they have no potential conflicts of interest.

                Article
                0074-02760170130
                10.1590/0074-02760170130
                5722262
                29236923
                02f6ecf6-192f-4369-9211-508e20dde476

                This is an Open Access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

                History
                : 02 April 2017
                : 20 June 2017
                Page count
                Figures: 2, Tables: 2, Equations: 0, References: 30, Pages: 7
                Categories
                Article

                leptospirosis,genomics,bioinformatics,next-generation sequencing,leptospira interrogans,zoonosis,phylogenetic analysis,whole-genome sequencing

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