豹纹鳃棘鲈致病性哈维氏弧菌的分离鉴定与系统发育分析  

Identification and phylogenetic analyses of Vibrio harveyi islated fromPlectropomusleopardus

作　　者：徐晓丽[1] 邵蓬[1] 李灏[1] 任涵玮 张勤[1] 

机构地区：[1]天津市水产技术推广站,天津300221

出　　处：《华中农业大学学报》2014年第4期112-118,共7页Journal of Huazhong Agricultural University

基　　金：天津市科技支撑计划项目(12ZCZDNC00900)

摘　　要：为探明养殖豹纹鳃棘鲈体表溃疡症的病因,从患病豹纹鳃棘鲈血液及内脏、体表溃疡处分离到2株优势菌,其中编号为1031的菌株经人工感染证实能够引起被感染鱼的死亡,为引起该豹纹鳃棘鲈溃疡病的病原菌。采用形态学、生理生化特性鉴定,结果表明菌株1031与弧菌属的基本特征相符,分别基于16SrDNA序列及热激蛋白60基因(hsp60)序列构建的系统发育树将菌株1031与哈维氏弧菌聚为一支,置信度均为97%,以1031为模板特异性扩增哈维氏弧菌toxR基因,也得到了预期382bp的核酸片段。综合以上结果,最终确定此次豹纹鳃棘鲈体表溃疡症的病原为哈维氏弧菌(Vibrio harveyi)。药敏结果显示,该菌株仅对氟苯尼考、庆大霉素(120μg)、菌必治敏感,对诺氟沙星、恩诺沙星、阿奇霉素、强力霉素等均不敏感。The aim of this study was to isolate the pathogens of ulceration on the body surface in Plectropomus leopardus. Two strains were isolated from the skin,blood,kidney and spleen of mortal P. leopardus. One strain coded 1031 could cause the mortality of healthy P. leopardus independently in ar- tificially infected experiment and were proved to be pathogen to ulceration of P. leopardus. Based on the results of morphological observation, physiological and biochemical characteristic analysis, strain 1031 was identified as Vibrio. 16S rDNA gene sequence further proved that strain 1031 belonged to V. har veyi. Phylogenetic analysis based on 16S rDNA gene and hsp60 gene indicated that strain 1031 showed the highest level of similarity to V. harveyi with bootstraps of 97%. The expected 382 bp-nucleic acid fragment of V. harveyi toxR gene was amplified using stain 1031 as a template. These results indicated that the pathogen of ulceration on the body surface in P. leopardus was V. harveyi. The antibiotic sensitivity analysis showed that strain 1031 was susceptible to florfenicol, gentamicinum （120 μg） and rocephin, but high resistant to norfloxacin, enrofloxacin, azithromycin, doxycycline, etc.

关 键 词：哈维氏弧菌 豹纹鳃棘鲈 16S RDNA HSP60 toxR 

分 类 号：S941.4[农业科学—水产养殖]