引用本文:张晓君,陈翠珍,阎斌伦,房 海,秦国民,徐 静.凡纳滨对虾(Litopenaeus vannamei)病原副溶血弧菌(Vibrio parahaemolyticus)的表型及分子特征.海洋与湖沼,2009,40(5):654-662.
【打印本页】   【HTML】   【下载PDF全文】   查看/发表评论  下载PDF阅读器  关闭
←前一篇|后一篇→ 过刊浏览    高级检索
本文已被:浏览 4581次   下载 4067 本文二维码信息
码上扫一扫!
分享到: 微信 更多
凡纳滨对虾(Litopenaeus vannamei)病原副溶血弧菌(Vibrio parahaemolyticus)的表型及分子特征
张晓君1, 陈翠珍2, 阎斌伦1, 房 海2, 秦国民1, 徐 静1
1.淮海工学院海洋学院 江苏省海洋生物技术重点建设实验室;2.河北科技师范学院动物科学系
摘要:
采用常规的形态及生理生化特性检验及分子生物学等方法, 对引起凡纳滨对虾幼虾发生毁灭性死亡的病原菌进行了致病性、表型生物学及分子生物学的系统研究。结果表明, 分离菌为弧菌属(Vibrio Pacini 1954)的副溶血弧菌(Vibrio parahaemolyticus), 对凡纳滨对虾、中国对虾及日本对虾仔虾均有强致病性。分离鉴定的4 株副溶血弧菌具有淀粉酶、明胶酶、卵磷脂酶活性, 但不具有蛋白酶、脂肪酶活性, KP 溶血均呈阴性, 且均具有K 抗原; 代表菌株(JGB080
关键词:  凡纳滨对虾, 副溶血弧菌, 16S rRNA 基因, gyrB 基因
DOI:10.11693/hyhz200905021021
分类号:
基金项目:淮海工学院江苏省海洋生物技术重点建设实验室研究基金资助, 2008HS016 号; 连云港市科技攻关项目资助, CN0826 号; 淮海工学院引进人才科研启动基金资助项目, KQ07106 号
PHENOTYPIC AND MOLECULAR CHARACTERIZATION OF PATHOGENIC VIBRIO PARAHAEMOLYTICUS ISOLATED FROM PENAEUS VANNAMEI
ZHANG Xiao-Jun1, CHEN Cui-Zhen2, YAN Bin-Lun1, FANG Hai2, QIN Guo-Min1, XU Jing1
1.Key Laboratory of Oceanic Biotechnology of Jiangsu, Huaihai Institute of Technology;2.Department of Animal Science, Hebei Normal University of Science and Technology
Abstract:
A mass mortality of cultured Litopenaeus vannamei broke out during summer in a local farm in northern Jiangsu Province. Syndromes include swimming near water surface and the pond walls, with black gill and opaque muscle. Tissue samples were taken in the liver, and strong pathogenicity to L. vannamei, P. japonicus and P. chinensis were determined by immersion. The phenotypic characteristics of isolates (4 strains) were examined in morphology, physiology and biochemistry; the 16S rRNA and gyrB were amplified by PCR and compared with sequences in databases, from which molecular phylogenetic trees were constructed. The sequenced 16S rRNA gene of strain JGB080708-1 (GenBank accession No.GQ205448) is 1456bp in length, the sequenced gyrB gene of strain JGB080708-1 (GenBank accession No.GQ205453) is 1195bp in length. Results of phylogenetic analysis on 16S rRNA and gyrB genes revealed a high homogeneity between the isolates and V. parahaemolyticus from GenBank database. Comparison in method between 16S rRNA and gyrB genes confirms that gyrB gene analysis is more useful. The isolates were identified as V. parahaemolyticus [(Fujino et al 1951) Sakazaki Nakamura and Takizawa 1963] of Vibrio (Pacini 1854) in phenotypic and molecular characteristics. Test on the activity of extracellulase and hemolysin shows that the isolates could produce diastase, gelatinase, and lecithinase, but no proteinase and lipase, and KP haemolysis was negative. Examination of K-antigen and agglutination titers show that the isolates have K-antigen, and the titer of OK-agglutinogen with OK-antiserum and O-agglutinogen with O-antiserum was the same (7log2); the titer of OK-antiserum with O-agglutinogen and O-antiserum with OK-agglutinogen was 2log2 and 3log2, respectively. The drug resistance of isolates to 49 antimicrobial agents was detected, the results show that the isolates are resistant to penicillin G, ampicillin, oxacillin, lincomycin, bacitracin and amoxicillin.
Key words:  Litopenaeus vannamei, Vibrio parahaemolyticus, 16S rRNA gene, gyrB gene
Copyright ©  海洋与湖沼 Oceanologia et Limnlolgia Sinica Copyright©2008 All Rights Reserved
Supervised by: 中国科协技术协会 Sponsored by: 中国海洋湖沼学会
Address: 青岛市福山路32号  Postcode: 266071  Tel: 0532-82898753  E-mail: ols@qdio.ac.cn
Technical support: Beijing E-Tiller Co.,Ltd.