创伤弧菌FJ03-X2胞外产物对欧洲鳗鲡的致病性和免疫原性分析  

Analysis of the Pathogenicity and Immunigenicity of Extracellular Products(ECPs) of Vibrio vulnificus FJ03-X2 to European Eels(Anguillia anguillia)

作　　者：董传甫[1] 林天龙[1] 俞伏松[1] 林能锋[1] 

机构地区：[1]福建省农业科学院生物技术研究所,福州350003

出　　处：《华中农业大学学报》2006年第1期71-75,共5页Journal of Huazhong Agricultural University

基　　金：福建省科技重大专项(2004NZ03-2);国家"863"项目(2003AA622050)资助

摘　　要：以70％饱和硫酸铵盐析法制备鳗源创伤弧菌生物1型高致病菌株FJ03-X2的胞外产物（ECPs），攻毒试验表明，该ECPs对鳗鲡无致病力；SDSPAGE分析表明，该ECPs由约36ku为主的系列蛋白条带组成。制备ECPs的免疫刺激复合物（ISCOMs），以20pg／尾的剂量腹腔注射免疫规格为15-20g／尾的欧洲鳗鲡，ELISA分析表明，21d和28d时的血清抗体效价约为1：1280，免疫印记显示，ECPs中分子量为36ku以上的蛋白成份可被免疫欧洲鳗鲡血清所识别，是构成ECPs的主要免疫原；免疫保护试验表明，免疫欧洲鳗鲡显示出较高的免疫保护力。试验结果也显示出，单纯以ECPs等剂量腹腔注射免疫欧洲鳗鲡，不能激发欧洲鳗鲡产生较高滴度的血清抗体，免疫欧洲鳗鲡也未能产生有效的保护性免疫应答。Extracellular products（ECPs） of FJ03-X2 strain isolated from the blood of the diseased eels and with the 50% lethal dosage （LD50） to eels （15-20 g per fish） about 7. 1 × 10^3 cfu per fish,were prepared. SDS-PAGE analysis exhibited that the ECPs consist of more than ten protein bands with the major protein band about 36 kilodalton. Given a dosage of 120 ug per fish , no obvious pathological changes were observed. ECPs of FJ03-X2 were not the direct pathogenic factors to eels. To enhance the immunogenicity of ECPs, ECPs-LISCOMs （Immune-stimulating complex, ISCOMs） were prepared by dialysis assay. Transmission electron micrograph showed that ECPs ISCOMs prepared had some basic matrix profiles contrast to the typical ISCOMs. European eels （Anguillia anguillia ） were immunized by intraperitoneal injection （i. p） at a dose of 20 ug per fish with both ECPs-ISCOMs and ECPs, respectively. Serum were collected on twenty-first day and twenty-eighth day. The average serum antibody titers immunized with ECPs-ISCOMs were about 1 ： 1 280 by enzyme linked immunosorbent assay （ELISA）. Immunoblot indicated that only the protein bands with the molecular weight over 36 ku could be recog nized by the anti-serum. Challenge experiment showed that eels immunized with ECPs-ISCOMs gained a high relative percent survival（RPS）. But no protective effect were observed when immunized with only ECPs.

关 键 词：创伤弧菌 欧洲鳗鲡 胞外产物 致病性 免疫原性 

分 类 号：Q936[生物学—微生物学] S941.42[农业科学—水产养殖]