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      Hybrid Vibrio cholerae El Tor Lacking SXT Identified as the Cause of a Cholera Outbreak in the Philippines

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          ABSTRACT

          Cholera continues to be a global threat, with high rates of morbidity and mortality. In 2011, a cholera outbreak occurred in Palawan, Philippines, affecting more than 500 people, and 20 individuals died. Vibrio cholerae O1 was confirmed as the etiological agent. Source attribution is critical in cholera outbreaks for proper management of the disease, as well as to control spread. In this study, three V. cholerae O1 isolates from a Philippines cholera outbreak were sequenced and their genomes analyzed to determine phylogenetic relatedness to V. cholerae O1 isolates from recent outbreaks of cholera elsewhere. The Philippines V. cholerae O1 isolates were determined to be V. cholerae O1 hybrid El Tor belonging to the seventh-pandemic clade. They clustered tightly, forming a monophyletic clade closely related to V. cholerae O1 hybrid El Tor from Asia and Africa. The isolates possess a unique multilocus variable-number tandem repeat analysis (MLVA) genotype (12-7-9-18-25 and 12-7-10-14-21) and lack SXT. In addition, they possess a novel 15-kb genomic island (GI-119) containing a predicted type I restriction-modification system. The CTXΦ-RS1 array of the Philippines isolates was similar to that of V. cholerae O1 MG116926, a hybrid El Tor strain isolated in Bangladesh in 1991. Overall, the data indicate that the Philippines V. cholerae O1 isolates are unique, differing from recent V. cholerae O1 isolates from Asia, Africa, and Haiti. Furthermore, the results of this study support the hypothesis that the Philippines isolates of V. cholerae O1 are indigenous and exist locally in the aquatic ecosystem of the Philippines.

          IMPORTANCE

          Genetic characterization and phylogenomics analysis of outbreak strains have proven to be critical for probing clonal relatedness to strains isolated in different geographical regions and over time. Recently, extensive genetic analyses of V. cholerae O1 strains isolated in different countries have been done. However, genome sequences of V. cholerae O1 isolates from the Philippines have not been available for epidemiological investigation. In this study, molecular typing and phylogenetic analysis of Vibrio cholerae isolated from both clinical and environmental samples in 2011 confirmed unique genetic features of the Philippines isolates, which are helpful to understand the global epidemiology of cholera.

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          Most cited references33

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          Performance comparison of benchtop high-throughput sequencing platforms.

          Three benchtop high-throughput sequencing instruments are now available. The 454 GS Junior (Roche), MiSeq (Illumina) and Ion Torrent PGM (Life Technologies) are laser-printer sized and offer modest set-up and running costs. Each instrument can generate data required for a draft bacterial genome sequence in days, making them attractive for identifying and characterizing pathogens in the clinical setting. We compared the performance of these instruments by sequencing an isolate of Escherichia coli O104:H4, which caused an outbreak of food poisoning in Germany in 2011. The MiSeq had the highest throughput per run (1.6 Gb/run, 60 Mb/h) and lowest error rates. The 454 GS Junior generated the longest reads (up to 600 bases) and most contiguous assemblies but had the lowest throughput (70 Mb/run, 9 Mb/h). Run in 100-bp mode, the Ion Torrent PGM had the highest throughput (80–100 Mb/h). Unlike the MiSeq, the Ion Torrent PGM and 454 GS Junior both produced homopolymer-associated indel errors (1.5 and 0.38 errors per 100 bases, respectively).
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            Cholera.

            Despite more than a century of study, cholera still presents challenges and surprises to us. Throughout most of the 20th century, cholera was caused by Vibrio cholerae of the O1 serogroup and the disease was largely confined to Asia and Africa. However, the last decade of the 20th century has witnessed two major developments in the history of this disease. In 1991, a massive outbreak of cholera started in South America, the one continent previously untouched by cholera in this century. In 1992, an apparently new pandemic caused by a previously unknown serogroup of V. cholerae (O139) began in India and Bangladesh. The O139 epidemic has been occurring in populations assumed to be largely immune to V. cholerae O1 and has rapidly spread to many countries including the United States. In this review, we discuss all aspects of cholera, including the clinical microbiology, epidemiology, pathogenesis, and clinical features of the disease. Special attention will be paid to the extraordinary advances that have been made in recent years in unravelling the molecular pathogenesis of this infection and in the development of new generations of vaccines to prevent it.
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              Cholera

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                Author and article information

                Journal
                mBio
                MBio
                mbio
                mbio
                mBio
                mBio
                American Society of Microbiology (1752 N St., N.W., Washington, DC )
                2150-7511
                21 April 2015
                Mar-Apr 2015
                : 6
                : 2
                : e00047-15
                Affiliations
                [ a ]St. Luke’s Medical Center, Research and Biotechnology Division, Quezon City, Philippines
                [ b ]Maryland Pathogen Research Institute, University of Maryland, College Park, Maryland, USA
                [ c ]CosmosID Inc., Rockville, Maryland, USA
                [ d ]Center for Bioinformatics and Computational Biology, University of Maryland, College Park, Maryland, USA
                [ e ]National Epidemiology Center, Department of Health, Manila, Philippines
                [ f ]Research Institute for Tropical Medicine, Alabang, Philippines
                [ g ]TMG Biosciences, LLC, Austin, Texas, USA
                [ h ]U.S. Army Edgewood Chemical Biological Center, Aberdeen, Maryland, USA
                [ i ]Department of Biology, Johns Hopkins University, Baltimore, Maryland, USA
                [ j ]Johns Hopkins Bloomberg School of Public Health, Baltimore, Maryland, USA
                Author notes
                Address correspondence to Rita R. Colwell, rcolwell@ 123456umiacs.umd.edu .
                [*]

                Present address: AITbiotech Pte Ltd., Science Park I, Singapore.

                [†]

                Deceased October 2014.

                Editor Anne K. Vidaver, University of Nebraska

                Article
                mBio00047-15
                10.1128/mBio.00047-15
                4453562
                25900650
                49c6c0f3-d3ea-4682-8f5b-212f96047041
                Copyright © 2015 Klinzing et al.

                This is an open-access article distributed under the terms of the Creative Commons Attribution-Noncommercial-ShareAlike 3.0 Unported license, which permits unrestricted noncommercial use, distribution, and reproduction in any medium, provided the original author and source are credited.

                History
                : 15 January 2015
                : 19 March 2015
                Page count
                supplementary-material: 0, Figures: 3, Tables: 1, Equations: 0, References: 33, Pages: 7, Words: 6379
                Categories
                Research Article
                Custom metadata
                March/April 2015

                Life sciences
                Life sciences

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