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      Genetic and pathogenic difference between Streptococcus agalactiae serotype Ia fish and human isolates

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          Abstract

          Background

          Streptococcus agalactiae (GBS) is a common pathogen to infect newborn, woman, the elderly, and immuno-compromised human and fish. 37 fish isolates and 554 human isolates of the GBS in 2007–2012 were investigated in serotypes, antibiotic susceptibility, genetic difference and pathogenicity to tilapia.

          Results

          PCR serotyping determined serotype Ia for all fish GBS isolates and only in 3.2 % (3–4.2 %) human isolates. For fish isolates, all consisted a plasmid less than 6 kb and belonged to ST7 type, which includes mainly pulsotypes I and Ia, with a difference in a deletion at the largest DNA fragment. These fish isolates were susceptible to all antimicrobials tested in 2007 and increased in non-susceptibility to penicillin, and resistance to clindamycin and ceftriaxone in 2011. Differing in pulsotype and lacking plasmid from fish isolates, human serotype Ia isolates were separated into eight pulsotypes II–IX. Main clone ST23 included pulsotypes II and IIa (50 %) and ST483 consisted of pulsotype III. Human serotype Ia isolates were all susceptible to ceftriaxone and penicillin and few were resistant to erythromycin, azithromycin, clindamycin, levofloxacin and moxifloxacine with the resistant rate of 20 % or less. Using tilapia to analyze the pathogenesis, fish isolates could cause more severe symptoms, including hemorrhage of the pectoral fin, hemorrhage of the gill, and viscous black and common scites, and mortality (>95 % for pulsotype I) than the human isolates (<30 %); however, the fish pulostype Ia isolate 912 with deletion caused less symptoms and the lowest mortality (<50 %) than pulsotype I isolates.

          Conclusion

          Genetic, pathogenic, and antimicrobial differences demonstrate diverse origin of human and fish serotype Ia isolates. The pulsotype Ia of fish serotype Ia isolates may be used as vaccine strains to prevent the GBS infection in fish.

          Electronic supplementary material

          The online version of this article (doi:10.1186/s12866-016-0794-4) contains supplementary material, which is available to authorized users.

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          Most cited references36

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          Rapid procedure for detection and isolation of large and small plasmids.

          Procedures are described for the detection and isolation of plasmids of various sizes (2.6 to 350 megadaltons) that are harbored in species of Agrobacterium, Rhizobium, Escherichia, Salmonella, Erwinia, Pseudomonas, and Xanthomonas. The method utilized the molecular characteristics of covalently closed circular deoxyribonucleic acid (DNA) that is released from cells under conditions that denature chromosomal DNA by using alkaline sodium dodecyl sulfate (pH 12.6) at elevated temperatures. Proteins and cell debris were removed by extraction with phenol-chloroform. Under these conditions chromosomal DNA concentrations were reduced or eliminated. The clarified extract was used directly for electrophoretic analysis. These procedures also permitted the selective isolation of plasmid DNA that can be used directly in nick translation, restriction endonuclease analysis, transformation, and DNA cloning experiments.
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            Multiplex PCR assay for rapid and accurate capsular typing of group B streptococci.

            We developed a simple, specific, and sensitive two-multiplex-PCR assay that enabled the detection of all known group B streptococcal (GBS) capsular polysaccharides. This test is well adapted for GBS capsular polysaccharide typing in large-scale epidemiological studies.
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              Aspects of the natural history and virulence of S. agalactiae infection in Nile tilapia.

              Streptococcus agalactiae is an emerging pathogen in Nile tilapia (Oreochromis niloticus) worldwide. To investigate aspects of the epidemiology, transmission and virulence of S. agalactiae infections, nine outbreaks of meningoencephalitis and septicemia in Nile tilapia farms in Brazil were analyzed. Records from the outbreaks revealed large variation in the weight of fish affected, high mortality, and disease occurrence at water temperatures above 26 degrees C. S. agalactiae was isolated from diseased fish from all farms, and 29 strains were identified by phenotypic tests and 16S rRNA gene sequencing. Five strains from different geographic origins were selected to determine the 50% lethal dose (LD(50)). All strains were highly virulent; for example, strain SA 20-06 had an LD(50) of 90 bacteria. To investigate S. agalactiae transmission, we conducted cohabitation assays with diseased and healthy fish and fish challenges using an immersion bath or gill inoculation. Strain SA 20-06 was used in all assays. The disease was reproduced with characteristic clinical signs and S. agalactiae was reisolated in all trials. The infection route studies were identified as by direct contact or through the water. In conclusion, S. agalactiae, a major pathogen of Nile tilapia in Brazil, exhibited high virulence, regardless of the geographic origin of the isolated strains.
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                Author and article information

                Contributors
                cschu@mail.ncyu.edu.tw
                s0933118@mail.ncyu.edu.tw
                chmhosp@cgmh.org.tw
                pedwang@adm.cgmh.org.tw
                dcba541@hotmail.com
                stu005301@hotmail.com
                chencc@mail.ncyu.edu.tw
                Journal
                BMC Microbiol
                BMC Microbiol
                BMC Microbiology
                BioMed Central (London )
                1471-2180
                2 August 2016
                2 August 2016
                2016
                : 16
                : 175
                Affiliations
                [1 ]Department of Microbiology, Immunology, and Biopharmaceutics, National Chiayi University, Chiayi, 60004 Taiwan, ROC
                [2 ]Department of Aquatic Biosciences, National Chiayi University, Chiayi, 60004 Taiwan, ROC
                [3 ]Department of Pediatrics, Chang Gung Memorial Hospital, Chiayi, Taiwan, ROC
                Article
                794
                10.1186/s12866-016-0794-4
                4971743
                27484120
                d9aded31-d654-476a-b639-6f4c7c823c35
                © The Author(s). 2016

                Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License ( http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver ( http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

                History
                : 16 April 2015
                : 29 July 2016
                Categories
                Research Article
                Custom metadata
                © The Author(s) 2016

                Microbiology & Virology
                streptococcosis,streptococcus agalactiae (gbs),pulsotype,mlst,serotype,tilapia
                Microbiology & Virology
                streptococcosis, streptococcus agalactiae (gbs), pulsotype, mlst, serotype, tilapia

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