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      Isolation and characterization of Streptococcus sp. from diseased flounder ( Paralichthys olivaceus) in Jeju Island

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          Abstract

          Streptococcus sp. is gram-positive coccus that causes streptococcal infections in fish due to intensification of aquaculture and caused significant economic losses in fish farm industry. A streptococcal infection occurred from cultured diseased olive flounder ( Paralichthys olivaceus) in May, 2005 at a fish farm in Jeju Island, Korea. The diseased flounder exhibited bilateral exophthalmic eyes and rotten gills; water temperature was 16~18℃ when samples were collected. Of the 22 fish samples collected, 3 samples were identified as Lactococcus garvieae and 18 samples were identified as Streptococcus parauberis by culture-based, biochemical test. Serological methods such as slide agglutination, hemolysis and antimicrobial susceptibility test were also used as well as multiplex PCR-based method to simultaneously detect and confirm the pathogens involved in the infection. S. parauberis and L. garvieae have a target region of 700 and 1100 bp., respectively. One fish sample was not identified because of the difference in the different biochemical and serological tests and was negative in PCR assay. In the present study, it showed that S. parauberis was the dominant species that caused streptococcosis in the cultured diseased flounder.

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          Development of a rapid and sensitive test for identification of major pathogens in bovine mastitis by PCR.

          Bovine mastitis is the most important source of loss for the dairy industry. A rapid and specific test for the detection of the main pathogens of bovine mastitis is not actually available. Molecular probes reacting in PCR with bacterial DNA from bovine milk, providing direct and rapid detection of Escherichia coli, Staphylococcus aureus, Streptococcus agalactiae, Streptococcus dysgalactiae, Streptococcus parauberis, and Streptococcus uberis, have been developed. Two sets of specific primers were designed for each of these microorganisms and appeared to discriminate close phylogenic bacterial species (e.g., S. agalactiae and S. dysgalactiae). In addition, two sets of universal primers were designed to react as positive controls with all major pathogens of bovine mastitis. The sensitivities of the test using S. aureus DNA extracted from milk with and without a pre-PCR enzymatic lysis step of bacterial cells were compared. The detection limit of the assay was 3.125 x 10(2) CFU/ml of milk when S. aureus DNA was extracted with the pre-PCR enzymatic step compared to 5 x 10(3) CFU/ml of milk in the absence of the pre-PCR enzymatic step. This latter threshold of sensitivity is still compatible with its use as an efficient tool of diagnosis in bovine mastitis, allowing the elimination of expensive reagents. The two PCR tests avoid cumbersome and lengthy cultivation steps, can be performed within hours, and are sensitive, specific, and reliable for the direct detection in milk of the six most prevalent bacteria causing bovine mastitis.
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            Molecular taxonomic studies on Streptococcus uberis types I and II. Description of Streptococcus parauberis sp. nov.

            The nucleotide sequences of 16S ribosomal-RNA of Streptococcus uberis types I and II were determined by reverse transcriptase. Comparative analysis of the sequence data showed that the two types are phylogenetically distinct and worthy of separate species status. A new species Streptococcus parauberis is proposed for the type II strains. The type strain of Strep. parauberis is NCDO 2020.
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              Differentiation of Lactococcus lactis and Lactococcus garvieae from humans by comparison of whole-cell protein patterns.

              We tested 12 reference and 24 clinical strains of lactococci for physiologic characteristics using a conventional test system, the Gen-Probe Enterococcus 2 chemiluminescence assay (Gen-Probe Inc., San Diego, Calif.), the Rapid Strep identification system (Analytab Products, Plainview, N.Y.), and whole-cell protein analysis. The Gen-Probe Enterococcus 2 chemiluminescence assay for Enterococcus identification was negative with all strains. Neither the conventional test nor the Rapid Strep identification system could differentiate between the two Lactococcus spp. most commonly isolated from humans. A simple procedure, based on sodium dodecyl sulfate-polyacrylamide gel electrophoresis, was developed for comparing the whole-cell protein patterns of Lactococcus spp. L. lactis and L. garvieae were differentiated by unique protein patterns.
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                Author and article information

                Journal
                J Vet Sci
                JVS
                Journal of Veterinary Science
                The Korean Society of Veterinary Science
                1229-845X
                1976-555X
                March 2006
                31 March 2006
                : 7
                : 1
                : 53-58
                Affiliations
                [1 ]Department of Oceanography, Pukyong National University, Busan 608-737, Korea.
                [2 ]College of Veterinary Medicine, Seoul National University, Seoul 151-742, Korea.
                [3 ]College of Fisheries and Aquatic Sciences, Iloilo State College of Fisheries, Iloilo, Philippines.
                [4 ]KRF Zoonotic Disease Priority Research Institute, Seoul National University, Seoul 151-742, Korea.
                Author notes
                Corresponding author: Tel: +82-2-880-1282; Fax: +82-2-880-1213, parksec@ 123456snu.ac.kr
                Article
                10.4142/jvs.2006.7.1.53
                3242086
                16434850
                459d6aa9-aefb-473c-a4a5-7a4e8012e879
                Copyright © 2006 The Korean Society of Veterinary Science
                History
                Categories
                Original Article

                Veterinary medicine
                lactococcus garvieae,streptococcus parauberis,multiplex pcr assay,streptococcus sp.

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