从患病黑鲪分离病原菌HV0811的鉴定及其系统发育分析  

Identification and phylogenetic analysis of pathogen HV0811 isolated from the diseased Sebastodes fuscescens

作　　者：孟鹏[1] 战文斌[1] 绳秀珍[1] 

机构地区：[1]中国海洋大学教育部海水养殖重点实验室,山东青岛266003

出　　处：《海洋科学》2010年第5期48-54,共7页Marine Sciences

基　　金：国家高技术研究发展计划(863)项目(2006AA100306);国家自然科学基金项目(30771648)

摘　　要：从症状为渍疡瞎眼的养殖黑鲪（Sebastodes fuscescens）病灶处分离到3株优势菌HV0811、PT0811和JH0811，以肌肉注射人工感染实验证实菌株HV0811为致病菌，其对黑鳕的半致死浓度为7．15×10^5CFu／尾。通过对致病菌的形态学观察、生理生化分析表明：菌株HV0811为革兰氏阴性菌，极生单鞭毛，短杆状，直径1～3mm，需Na＋生长，低于4℃、高于42℃不生长。基于16SrRNA和HSP60基因序列构建的系统发育树表明：HV0811分别与哈维氏弧菌（AY750578和AY332571）相类聚，其中16SrRNA基因序列与哈维氏弧菌（AY750578）聚合置信度较低，仅有10％，不能有效地确定该种；HSP60基因序列与哈维氏弧菌（AY332571）聚合为一个分支的置信度达100％。综合该菌的形态、生理生化及HSP60基因序列比对结果，表明分离到的病原菌HV0811为哈维氏弧菌。药敏试验结果显示病原哈维氏弧菌（HV0811）对庆大霉素、新霉素、氟哌酸等较为敏感。Bacterial strains HV0811, PT0811 and JH0811 were isolated from the Sebastodesfuscescens with skin ulceration and blind. Strain HV0811 was proved to be a pathogen by muscle injection, with the LD50 being 7.15 ×10^5 CFU per fish. By morphological observation and physiological-biochemical analysis, HV0811 was identified as gram-negative with single polar flagellum, in a shape of short rod and a diameter of 1-3mm; it could grow between 4℃ and 42℃, but it could not grow without Na＋. Phylogenetic tree of Vibrio based on 16S rRNA and HSP60 gene indicated that strain HV0811 clustered together with V.harveyi （AY750578） and （AY332571） respectively. The boot strap of strain HV0811 clustered together with V.harveyi （AY750578） based on 16S rRNA gene was only 10%, which was too low to give identification of the strain effectively; But strain HV0811 clustered together with V .harveyi （AY332571） based on HSP60 gene had a high boot strap of 100%. So, all above results confirmed that strain HV0811 was V.harveyi, Additionally, drug sensitivity screen using 30 antibacterial agents showed that strain HV0811 was sensitive to several antibacterial agents including Gentamicin, neomycin and Norfloxacin.

关 键 词：黑鲪(Sebastodes fuscescens) 溃疡 瞎眼 哈维氏弧菌(Vibrio harveyi) 16S RRNA基因 HSP60基因 

分 类 号：S943[农业科学—水产养殖]