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      Updating the Vibrio clades defined by multilocus sequence phylogeny: proposal of eight new clades, and the description of Vibrio tritonius sp. nov.

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          Abstract

          To date 142 species have been described in the Vibrionaceae family of bacteria, classified into seven genera; Aliivibrio, Echinimonas, Enterovibrio, Grimontia, Photobacterium, Salinivibrio and Vibrio. As vibrios are widespread in marine environments and show versatile metabolisms and ecologies, these bacteria are recognized as one of the most diverse and important marine heterotrophic bacterial groups for elucidating the correlation between genome evolution and ecological adaptation. However, on the basis of 16S rRNA gene phylogeny, we could not find any robust monophyletic lineages in any of the known genera. We needed further attempts to reconstruct their evolutionary history based on multilocus sequence analysis (MLSA) and/or genome wide taxonomy of all the recognized species groups. In our previous report in 2007, we conducted the first broad multilocus sequence analysis (MLSA) to infer the evolutionary history of vibrios using nine housekeeping genes (the 16S rRNA gene, gapA, gyrB, ftsZ, mreB, pyrH, recA, rpoA, and topA), and we proposed 14 distinct clades in 58 species of Vibrionaceae. Due to the difficulty of designing universal primers that can amplify the genes for MLSA in every Vibrionaceae species, some clades had yet to be defined. In this study, we present a better picture of an updated molecular phylogeny for 86 described vibrio species and 10 genome sequenced Vibrionaceae strains, using 8 housekeeping gene sequences. This new study places special emphasis on (1) eight newly identified clades ( Damselae, Mediterranei, Pectenicida, Phosphoreum, Profundum, Porteresiae, Rosenbergii, and Rumoiensis); (2) clades amended since the 2007 proposal with recently described new species; (3) orphan clades of genomospecies F6 and F10; (4) phylogenetic positions defined in 3 genome-sequenced strains (N418, EX25, and EJY3); and (5) description of V. tritonius sp. nov., which is a member of the “ Porteresiae” clade.

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          Multilocus sequence typing: a portable approach to the identification of clones within populations of pathogenic microorganisms.

          Traditional and molecular typing schemes for the characterization of pathogenic microorganisms are poorly portable because they index variation that is difficult to compare among laboratories. To overcome these problems, we propose multilocus sequence typing (MLST), which exploits the unambiguous nature and electronic portability of nucleotide sequence data for the characterization of microorganisms. To evaluate MLST, we determined the sequences of approximately 470-bp fragments from 11 housekeeping genes in a reference set of 107 isolates of Neisseria meningitidis from invasive disease and healthy carriers. For each locus, alleles were assigned arbitrary numbers and dendrograms were constructed from the pairwise differences in multilocus allelic profiles by cluster analysis. The strain associations obtained were consistent with clonal groupings previously determined by multilocus enzyme electrophoresis. A subset of six gene fragments was chosen that retained the resolution and congruence achieved by using all 11 loci. Most isolates from hyper-virulent lineages of serogroups A, B, and C meningococci were identical for all loci or differed from the majority type at only a single locus. MLST using six loci therefore reliably identified the major meningococcal lineages associated with invasive disease. MLST can be applied to almost all bacterial species and other haploid organisms, including those that are difficult to cultivate. The overwhelming advantage of MLST over other molecular typing methods is that sequence data are truly portable between laboratories, permitting one expanding global database per species to be placed on a World-Wide Web site, thus enabling exchange of molecular typing data for global epidemiology via the Internet.
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            Digital DNA-DNA hybridization for microbial species delineation by means of genome-to-genome sequence comparison

            The pragmatic species concept for Bacteria and Archaea is ultimately based on DNA-DNA hybridization (DDH). While enabling the taxonomist, in principle, to obtain an estimate of the overall similarity between the genomes of two strains, this technique is tedious and error-prone and cannot be used to incrementally build up a comparative database. Recent technological progress in the area of genome sequencing calls for bioinformatics methods to replace the wet-lab DDH by in-silico genome-to-genome comparison. Here we investigate state-of-the-art methods for inferring whole-genome distances in their ability to mimic DDH. Algorithms to efficiently determine high-scoring segment pairs or maximally unique matches perform well as a basis of inferring intergenomic distances. The examined distance functions, which are able to cope with heavily reduced genomes and repetitive sequence regions, outperform previously described ones regarding the correlation with and error ratios in emulating DDH. Simulation of incompletely sequenced genomes indicates that some distance formulas are very robust against missing fractions of genomic information. Digitally derived genome-to-genome distances show a better correlation with 16S rRNA gene sequence distances than DDH values. The future perspectives of genome-informed taxonomy are discussed, and the investigated methods are made available as a web service for genome-based species delineation.
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              Report of the ad hoc committee for the re-evaluation of the species definition in bacteriology.

              An ad hoc committee for the re-evaluation of the species definition in bacteriology met in Gent, Belgium, in February 2002. The committee made various recommendations regarding the species definition in the light of developments in methodologies available to systematists.
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                Author and article information

                Journal
                Front Microbiol
                Front Microbiol
                Front. Microbiol.
                Frontiers in Microbiology
                Frontiers Media S.A.
                1664-302X
                18 October 2013
                27 December 2013
                2013
                : 4
                : 414
                Affiliations
                [1] 1Laboratory of Microbiology, Faculty of Fisheries Sciences, Hokkaido University Hakodate, Japan
                [2] 2Division of Genomics and Bioenvironmental Science, Frontier Science Research Center, University of Miyazaki Miyazaki, Japan
                [3] 3Department of Food and Nutrition, Hakodate Junior College Hakodate, Japan
                [4] 4National Institute for Interdisciplinary Science and Technology (CSIR) Kerala, India
                [5] 5National Research Institute of Fisheries Science, Fisheries Research Agency Yokohama, Japan
                [6] 6Department of Genetics, Center of Health Sciences, Federal University of Rio de Janeiro (UFRS) Rio de Janeiro, Brazil
                [7] 7A.C. Unidad Mazatlán, CIAD Mazatlán, México
                [8] 8CNRS UMR 7138, Systématique-Adaptation-Evolution Nice, France
                [9] 9Systématique-Adaptation-Evolution, Université de Nice-Sophia Antipolis Nice, France
                [10] 10Graduate School of Medical and Dental Sciences, Tokyo Medical and Dental University Tokyo, Japan
                [11] 11Earth-Life Science Institute, Tokyo Institute of Technology Tokyo, Japan
                Author notes

                Edited by: Daniela Ceccarelli, University of Maryland, USA

                Reviewed by: Xiu-Lan Chen, Shandong University, China; Matteo Spagnoletti, University College London, UK

                *Correspondence: Tomoo Sawabe, Laboratory of Microbiology, Faculty of Fisheries Sciences, Hokkaido University, 3-1-1 Minato-cho, Hakodate 041-8611, Japan e-mail: sawabe@ 123456fish.hokudai.ac.jp

                This article was submitted to Aquatic Microbiology, a section of the journal Frontiers in Microbiology.

                Article
                10.3389/fmicb.2013.00414
                3873509
                24409173
                28ad51d5-c675-469e-aedb-acfc5f9c064e
                Copyright © 2013 Sawabe, Ogura, Matsumura, Feng, Amin, Mino, Nakagawa, Sawabe, Kumar, Fukui, Satomi, Matsushima, Thompson, Gomez-Gil, Christen, Maruyama, Kurokawa and Hayashi.

                This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

                History
                : 27 September 2013
                : 16 December 2013
                Page count
                Figures: 4, Tables: 4, Equations: 0, References: 51, Pages: 14, Words: 9500
                Categories
                Microbiology
                Original Research Article

                Microbiology & Virology
                vibrios,vibrionaceae,housekeeping protein gene,multilocus sequence analysis,evolution,vibrio tritonius

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