Barnes Andrew C, Young Fiona M, Horne Michael T, Ellis Anthony E
Fisheries Research Services Marine Laboratory, Victoria Road, Aberdeen AB11 9DB, Scotland, UK.
Dis Aquat Organ. 2003 Feb 27;53(3):241-7. doi: 10.3354/dao053241.
The biochemical profiles, presence of capsule, outer membrane protein profiles and serological interactions of isolates of Streptococcus iniae obtained from different geographical and fish host origins were examined. The isolates had very similar biochemical profiles using API 20 Strep but varied as to whether they were arginine dihydrolase-negative, -positive or -intermediate (AD-ve, AD+ve, AD+/-ve, respectively). Representatives of each AD type were compared in subsequent experiments. All types possessed a polysaccharide capsule. Sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) analysis of outer membrane proteins or whole cells revealed no difference in banding patterns between isolates. All isolates were resistant to trout normal and specific immune serum and grew well in the presence of added fresh normal serum. Serological analyses of the isolates revealed antigenic differences. Trout antiserum against the AD+ve isolate did not agglutinate the AD-ve or AD+/-ve isolates, while antisera against the latter 2 types showed low agglutinating activity with all 3 isolates. When whole live cells of AD-ve and AD+ve isolates were dot-blotted, antiserum to the AD+ve isolate did not stain the AD-ve isolate, but antiserum to the AD-ve isolate stained both AD types. However, if the cells were pre-treated with Proteinase K (to remove surface-exposed protein antigens), the AD+ve isolate was stained only by its homologous antiserum. These results suggest that while certain protein antigens of the different AD type strains are immunologically cross-reactive, the capsular antigens appear to be AD type-specific. Furthermore, the results suggest that the cross-reactive antigens on the AD-ve isolate are effectively hidden by the strain-specific capsule, while they are partially exposed on the AD+ve isolate.
对从不同地理区域和鱼类宿主来源获得的海豚链球菌分离株的生化特征、荚膜的存在、外膜蛋白图谱和血清学相互作用进行了研究。使用API 20 Strep检测时,这些分离株具有非常相似的生化特征,但在精氨酸双水解酶阴性、阳性或中间型(分别为AD-ve、AD+ve、AD+/-ve)方面存在差异。在后续实验中对每种AD类型的代表菌株进行了比较。所有类型都具有多糖荚膜。对分离株的外膜蛋白或全细胞进行十二烷基硫酸钠聚丙烯酰胺凝胶电泳(SDS-PAGE)分析,结果显示各分离株之间的条带模式没有差异。所有分离株均对虹鳟正常血清和特异性免疫血清具有抗性,并且在添加新鲜正常血清的情况下生长良好。对分离株的血清学分析显示存在抗原差异。针对AD+ve分离株的虹鳟抗血清不能凝集AD-ve或AD+/-ve分离株,而针对后两种类型的抗血清对所有三种分离株的凝集活性较低。当对AD-ve和AD+ve分离株的全活细胞进行斑点印迹时,针对AD+ve分离株的抗血清不能使AD-ve分离株染色,但针对AD-ve分离株的抗血清可使两种AD类型的菌株染色。然而,如果细胞先用蛋白酶K处理(以去除表面暴露的蛋白质抗原),则AD+ve分离株仅被其同源抗血清染色。这些结果表明,虽然不同AD类型菌株的某些蛋白质抗原具有免疫交叉反应性,但荚膜抗原似乎是AD类型特异性的。此外,结果表明AD-ve分离株上的交叉反应性抗原被菌株特异性荚膜有效隐藏,而在AD+ve分离株上则部分暴露。