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Nho, Seong-Won,Shin, Gee-Wook,Park, Seong-Bin,Jang, Ho-Bin,Cha, In-Seok,Ha, Mi-Ae,Kim, Young-Rim,Park, Yon-Kyoung,Dalvi, Rishikesh S.,Kang, Bong-Jo,Joh, Seong-Joon,Jung, Tae-Sung Oxford University Press 2009 FEMS microbiology letters Vol.293 No.1
<P>The etiological agents of streptococcosis were isolated from diseased olive flounder collected on the Jeju island of Korea. A total of 151 bacterial isolates were collected between 2003 and 2006. The isolates were examined using various phenotypic and proteomic analyses, including sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), immunoblotting, and glycoprotein assays. In addition, isolates were grown on blood agar to assess hemolytic activity, and biochemical assays were performed using the API20 Strep kit. Our results revealed that all isolates were nonmotile, Gram-positive cocci that displayed negative catalase and oxidase activities. Multiplex PCR assays revealed that 43% and 57% of the isolates were Streptococcus iniae and Streptococcus parauberis, respectively. These results were consistent with those of the SDS-PAGE and immunoblot analyses using whole-cell lysates of bacterial isolates. Significant differences were observed with respect to the Voges-Proskauer, pyrrodonyl arylamidase, alkaline phosphatase, and hemolytic activities of the S. iniae and S. parauberis isolates. Isolates of S. iniae displayed uniform profiles in the immunoblot and glycoprotein assays; however, immunoblot assays of S. parauberis isolates (using a chicken IgY antibody raised against a homologous isolate) revealed three distinct antigenic profiles. Our findings suggest that S. parauberis and S. iniae are endemic pathogens responsible for the development of streptococcosis in olive flounder.</P>
Shin, Gee-Wook,Nho, Seong-Won,Park, Seong-Bin,Jang, Ho-Bin,Cha, In-Seok,Ha, Mi-Ae,Kim, Young-Rim,Dalvi, Rishikesh S.,Joh, Seong-Joon,Jung, Tae-Sung Elsevier 2009 Veterinary microbiology Vol.139 No.1
<P><B>Abstract</B></P><P><I>Lactococcus garvieae</I> is an important etiological agent of lactococcosis in various fish species including olive flounder (<I>Paralichthys olivaceus</I>). In this study, proteomic and immunoproteomic analyses were employed to compare the antigenic profiles of strains KG9408, MS93003, and NSS9310 strains of <I>L. garvieae</I>. Proteomic analysis using two-dimensional gel electrophoresis (2-DE) revealed differences in five protein spots among the different <I>L. garvieae</I> strains. In immunoproteomic analysis, there was a significant difference in the 2-DE immunoblot profiles of the <I>L. garvieae</I> strains using sera collected from fish surviving infection with either <I>L. garvieae</I> strains KG9408 or NSS9310. These sera reacted with 8 and 7 unique antigenic protein spots, respectively. Heat shock protein (HSP) 70 and DNA-directed RNA polymerase were among the specific antigens recognized by the anti-NSS9310 serum. In addition, the anti-NSS9310 and anti-KG9408 olive flounder sera reacted with 25 common antigenic protein spots of all the <I>L. garvieae</I> strains, which included elongation factor (EF)-Tu, arginine deiminase (AD), inosine-5′-monophosphate dehydrogenase (IMPD), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), phosphomannomutase (PMM), <SMALL>L</SMALL>-lactate dehydrogenase (<SMALL>L</SMALL>-LDH), 6-phosphofructokinase and UDP-galactose 4-epimerase (UDP-galactose). Based on the present results, the 8 antigens recognized by the anti-KG9408 serum and the 25 common antigens recognized by both sera may serve as potential markers for developing an effective vaccine against this bacterium.</P>