【作者】 沈锦玉； 余旭平； 曹铮； 沈琴芳； 尹文林； 潘晓艺； 吴颖蕾；

【Author】 SHEN Jin-yu~1 YU Xu-ping~2 Cao Zheng~1 SHEN Qin-fang~2 YIN Wen-lin~1 PAN Xiao-yi~1 WU Ying-lei~1 (1.Zhejiang Institute of Freshwater Fisheries,Huzhou 313001,China;2.Zhejiang University,Hangzhou 310029,China)

【机构】 浙江省淡水水产研究所； 浙江大学；

【摘要】 浙江某地深水网箱养殖大黄鱼发生大量死亡,从病鱼体内分离出致病力较强的菌株GYC1108-1,人工感染试验证实为大黄鱼的病原菌。对细菌进行了形态、生理生化特性的测定,并进行了16S rRNA分子鉴定。PCR扩增获得了GYC1108-1菌株大小约1.5Kb的16S rDNA部分基因片段,产物经回收纯化,克隆到pMD18-T载体,转化大肠杆菌TG1,对阳性克隆子进行酶切及PCR鉴定,并测序,将测定的序列递交NCBI进行BLAST同源序列比对,与溶藻弧菌的标准菌株的16S rDNA具有高的同源性。结合菌株的生理生化特性,GYC1108-1菌株属于溶藻弧菌。并测定了其对14种药物的敏感性。同批病鱼肝、脾、肾组织滤液注射健康鱼后,也导致鱼的死亡:电镜观察到组织中存在50～70nm大小的病毒样颗粒。更多还原

【Abstract】 A serious epdemic of Great Yellow Croaker cultured in netcage in zhejiang was broken out in Sep.to Oct.in 2003 and 2004.The strain GYC1108-1 isolated from the diseased fish was confirmed to be the pathogen of the epidemic by artificial infection test.The colony morphological,physiological and biochemical characters of GYC1108-1 isolate was determined.About 1.5Kb fragments of the isolate was amplified by PCR using general primers of nuclear ribosomal DNA.The PCR product was extracted and cloned into pMD 18-T vector.After the transformation into E.coli TG1,the recombination plasmid was identified by restriction enzymes digestion and by PCR.One selected clone was then sequenced.The BLAST results of the sequence showed that it has high homology with respect sequence of standard strain of Vibrio alginotyticus.The results indicated that GYC1108-1 isolate belong to Vibrio alginolyticus. Meanwhile the sensitivity of strain to 14 kinds of antibiotic was detected.The healthy Great Yellow Croaker after injected with filtered fluid from ground tissues of diseased Great Yellow Croaker appeared the same symptom as that of the natural diseased one.Examination by electron microscopy showed that there were a numbers of viral particles in the tissues of spleen,kidney and liver.更多还原