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      Isolation and Characterization of Numerous Novel Phages Targeting Diverse Strains of the Ubiquitous and Opportunistic Pathogen Achromobacter xylosoxidans

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          Abstract

          The clinical relevance of nosocomially acquired infections caused by multi-resistant Achromobacter strains is rapidly increasing. Here, a diverse set of 61 Achromobacter xylosoxidans strains was characterized by MultiLocus Sequence Typing and Phenotype MicroArray technology. The strains were further analyzed in regard to their susceptibility to 35 antibiotics and to 34 different and newly isolated bacteriophages from the environment. A large proportion of strains were resistant against numerous antibiotics such as cephalosporines, aminoglycosides and quinolones, whereas piperacillin-tazobactam, ticarcillin, mezlocillin and imipenem were still inhibitory. We also present the first expanded study on bacteriophages of the genus Achromobacter that has been so far a blank slate with respect to phage research. The phages were isolated mainly from several waste water treatment plants in Germany. Morphological analysis of all of these phages by electron microscopy revealed a broad diversity with different members of the order Caudovirales, including the families Siphoviridae, Myoviridae, and Podoviridae. A broad spectrum of different host ranges could be determined for several phages that lysed up to 24 different and in part highly antibiotic resistant strains. Molecular characterisation by DNA restriction analysis revealed that all phages contain linear double-stranded DNA. Their restriction patterns display distinct differences underlining their broad diversity.

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          eBURST: inferring patterns of evolutionary descent among clusters of related bacterial genotypes from multilocus sequence typing data.

          The introduction of multilocus sequence typing (MLST) for the precise characterization of isolates of bacterial pathogens has had a marked impact on both routine epidemiological surveillance and microbial population biology. In both fields, a key prerequisite for exploiting this resource is the ability to discern the relatedness and patterns of evolutionary descent among isolates with similar genotypes. Traditional clustering techniques, such as dendrograms, provide a very poor representation of recent evolutionary events, as they attempt to reconstruct relationships in the absence of a realistic model of the way in which bacterial clones emerge and diversify to form clonal complexes. An increasingly popular approach, called BURST, has been used as an alternative, but present implementations are unable to cope with very large data sets and offer crude graphical outputs. Here we present a new implementation of this algorithm, eBURST, which divides an MLST data set of any size into groups of related isolates and clonal complexes, predicts the founding (ancestral) genotype of each clonal complex, and computes the bootstrap support for the assignment. The most parsimonious patterns of descent of all isolates in each clonal complex from the predicted founder(s) are then displayed. The advantages of eBURST for exploring patterns of evolutionary descent are demonstrated with a number of examples, including the simple Spain(23F)-1 clonal complex of Streptococcus pneumoniae, "population snapshots" of the entire S. pneumoniae and Staphylococcus aureus MLST databases, and the more complicated clonal complexes observed for Campylobacter jejuni and Neisseria meningitidis.
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            LIAN 3.0: detecting linkage disequilibrium in multilocus data. Linkage Analysis.

            LIAN is a program to test the null hypothesis of linkage equilibrium for multilocus data. LIAN incorporates both a Monte Carlo method as well as a novel algebraic method to carry out the hypothesis test. The program further returns the genetic diversity of the sample and the pairwise distances between its members.
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              Fix the antibiotics pipeline.

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                Author and article information

                Contributors
                Role: Editor
                Journal
                PLoS One
                PLoS ONE
                plos
                plosone
                PLoS ONE
                Public Library of Science (San Francisco, USA )
                1932-6203
                2014
                22 January 2014
                : 9
                : 1
                : e86935
                Affiliations
                [1 ]Leibniz Institute DSMZ – German Collection of Microorganisms and Cell Cultures, Department of Microorganisms, Braunschweig, Germany
                [2 ]Department of Microbiology/Genetechnology, Faculty of Biology, University of Bielefeld, Bielefeld, Germany
                [3 ]Helmholtz Centre for Infection Research, Department of Medical Microbiology, Central Facility for Microscopy, Braunschweig, Germany
                [4 ]Leibniz Institute DSMZ – German Collection of Microorganisms and Cell Cultures, Department of Microbial Ecology and Diversity Research, Braunschweig, Germany
                Charité-University Medicine Berlin, Germany
                Author notes

                Competing Interests: The authors have declared that no competing interests exist.

                Conceived and designed the experiments: JW CR JS. Performed the experiments: JW BD MR. Analyzed the data: JW JS. Wrote the paper: JW JS.

                Article
                PONE-D-13-19861
                10.1371/journal.pone.0086935
                3899368
                24466294
                e2fb6846-7809-4bdc-a984-639c642b5e0f
                Copyright @ 2014

                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 author and source are credited.

                History
                : 14 May 2013
                : 19 December 2013
                Page count
                Pages: 12
                Funding
                This work was supported by Leibniz Institute DSMZ - German Collection of Microorganisms and Cell Cultures funded this research through the employment of Johannes Wittmann, Christine Rohde and Johannes Sikorski, who contributed to all aspects of the study. There are no external funders who could have had a role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
                Categories
                Research Article
                Biology
                Ecology
                Biodiversity
                Freshwater Ecology
                Microbial Ecology
                Microbiology
                Bacterial Pathogens
                Gram Negative
                Virology
                Viral Classification
                DNA viruses
                Viral Transmission and Infection
                Virulence Factors and Mechanisms
                Emerging Infectious Diseases
                Medical Microbiology
                Microbial Control
                Microbial Ecology
                Microbial Physiology

                Uncategorized
                Uncategorized

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