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
Laboratory of Aquatic Animal Diseases, Research Institute of Life Science, College of Veterinary medicine, Gyeongsang National University, Jinju, Gyeongnam, Korea.
FEMS Microbiol Lett. 2009 Apr;293(1):20-7. doi: 10.1111/j.1574-6968.2009.01491.x.
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.
从韩国济州岛采集的患病牙鲆中分离出链球菌病的病原体。2003年至2006年间共收集了151株细菌分离株。使用各种表型和蛋白质组学分析方法对这些分离株进行检测,包括十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)、免疫印迹和糖蛋白测定。此外,将分离株接种在血琼脂上以评估溶血活性,并使用API20 Strep试剂盒进行生化检测。我们的结果显示,所有分离株均为无动力、革兰氏阳性球菌,过氧化氢酶和氧化酶活性呈阴性。多重PCR检测显示,分别有43%和57%的分离株为海豚链球菌和副乳房链球菌。这些结果与使用细菌分离株全细胞裂解物进行的SDS-PAGE和免疫印迹分析结果一致。在海豚链球菌和副乳房链球菌分离株的Voges-Proskauer、吡咯烷酮芳基酰胺酶、碱性磷酸酶和溶血活性方面观察到显著差异。海豚链球菌分离株在免疫印迹和糖蛋白测定中显示出一致的图谱;然而,副乳房链球菌分离株的免疫印迹分析(使用针对同源分离株产生的鸡IgY抗体)显示出三种不同的抗原图谱。我们的研究结果表明,副乳房链球菌和海豚链球菌是导致牙鲆链球菌病发生的地方性病原体。