半滑舌鳎弧菌病病原的分离鉴定  

Isolation and identification of pathogen responsible for the vibriosis in cultured Cynoglossus semilaevisis

作　　者：徐晓丽[1] 叶红梅[1] 侯纯强[2] 蔡琰[1] 钟文慧[1] 李贺密 XU Xiao-li YE Hong-mei HOU Chun-qiang CAI Yan ZHONG Wen-hui(Tianjin Fishery Technical Extension Department, Tianjin 300221, China Tianjin Bohai Sea Fisheries Research Institute, Tianjin 300457, China)

机构地区：[1]天津市水产技术推广站,天津河西300221 [2]天津渤海水产研究所,天津经济技术开发区300457

出　　处：《中国兽医杂志》2017年第8期86-90,共5页Chinese Journal of Veterinary Medicine

基　　金：天津市科委重大科技工程计划项目(14ZCDGNC00100);天津市水产局科研推广项目(J2016-03)

摘　　要：从患病半滑舌鳎(Cynoglossus semilaevis)内脏中分离到2株优势菌株,人工感染试验证实菌株150803对半滑舌鳎具有致病性;采用形态学观察、生理生化特性分析、16S r RNA等方法对所分离菌株进行鉴定,结果显示,菌株150803为革兰阴性杆菌,生化特性与溶藻弧菌(Vibrio alginolyticus)较为接近,以hsp60基因为遗传标记构建的系统发育树将菌株150803与溶藻弧菌聚为一支,置信度为99%,采用特异性引物扩增溶藻弧菌dnaJ基因得到预期144 bp的基因片段。综合以上结果判定引起此次半滑舌鳎发病的病原为溶藻弧菌。对22种抗菌药物的敏感性试验表明,菌株150803对恩诺沙星、环丙沙星、庆大霉素、菌必治敏感,对氟苯尼考、呋喃唑酮、苯唑青霉素等具有抗性。Dominant bacteria designed as 150802 and 150803 were isolated from the internal of diseasedCynoglossus semilaevisis and the stain 150803 was proved to be highly pathogenic toC. semilaevisisby artificial challenge. The biological characteristics inclu-ding morphological, physiological and bio-chemical characteristics were examined, and the hsp60 gene was sequenced and com-pared with the sequences deposited in Gen Bank databases. The results showed that the strain 150803 was Gram-negative bacilli.Based on the physiological and biochemical characteristic analysis, strain 150803 was identified asVibrio alginolyticus. Phylogenetictrees of Vibrio based on hsp60 gene indicated that strain 150803 showed the highest level of similarity toV. alginolyticusand thebootstrap was 99%. The expected 144 bp-nucleic acid fragment ofV. alginolyticus dna Jgene was amplified with the stain 150803 as a template. These results demonstrated that the vibriosis pathogen causing mortality inC. semilaevisiswasV. alginolyticus. Anti-microbial susceptibility assay showed that strain 150803 was susceptible to enrofloxacin, ciprofloxacin, gentamicinum and rocephin,but resistant to florfenicol, furazolidone, oxacillin, etc.

关 键 词：半滑舌鳎 溶藻弧菌 HSP60 DNAJ 

分 类 号：S941.417[农业科学—水产养殖]