| 摘要: |
| 采用表观分类学及分子生物学方法,对从大菱鲆细菌性败血感染症的病(死)鱼中分离到的相应病原菌,进行了形态特征、理化特性等较系统的鉴定及代表菌株DNA中G+Cmol%的测定;同时,择代表菌株进行了16SrRNA基因的分子鉴定,测定了16SrRNA基因序列、分析了相关细菌相应序列的同源性、构建了系统发生树。结果表明,所检30株菌均为利斯顿氏菌属(Listonella MacDonell and Colwell 1986)的鳗利斯顿氏菌[L. anguillarum (Bergeman 1909) MacDonell and Colwell 1986],择S010610-1株、S010610-3株及S010623-1株作为代表菌株进行的16SrRNA基因序列测定,不包括引物结合区,所扩增的16SrRNA基因序列长度,S010610-1株为1466bp(GenBank登录号:AY963630)、S010610-3株为1416bp(GenBank登录号:AY963631)、S010623-1株为1424bp(GenBank登录号:AY963632),与GenBank数据库中弧菌属细菌的同源性在97%—99%。 |
| 关键词: 大菱鲆 鳗利斯顿氏菌 鉴定 系统发育学分析 |
| DOI： |
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| 基金项目:河北省科技厅科技攻关项目,02220502D号;河北省自然科学基金项目,C2005000498号 |
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| IDENTIFICATION OF PATHOGEN LISTONELLA ANGUILLARUM ISOLATED FROM TURBOT SCOPHTHALMUS MAXIMUS |
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ZHANG Xiao-Jun1, CHEN Cui-Zhen1, FANG Hai1, ZHAN Wen-Bin2, JIN Xiao-Min1, WANG Xiu-Yun1
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1.Department of Animal Science, Hebei Normal University of Science and Technology, Qinhuangdao, 066600;2.Laboratory of Pathology and Immunology of Aquatic Animals, the Key Laboratory of Mariculture, Ministry of Education, Ocean University of China,Qingdao, 266003
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| Abstract: |
| Turbot (Scopthalamus maximus L.) is a fish species in high economic value aquacultured widely in northern China. However, cases of epizootic diseases of turbot were reported in growing number due to pathogenic bacteria infections often causing frequent mass mortality resulted from high density stocking. For alleviating the problem in search for the solution, some infected moribund turbots were collected from a farm in Laizhou, Shandong Province, July 2001, from which pathogenic bacteria were isolated. In this paper, the isolation and characterization of the bacteria are described and identified in morphology, physiology, biochemistry and bioinformics. Representative strains (S010610-1, S010610-3 and S010623-1) were selected for molecular identification by partial 16S rRNA gene sequencing and a molecular phylogenetic tree was constructed.
30 strains were examined and found to be quite consistent in morphology, being Gram-negative and rod-shaped, single or paired with round ends, no endospores, and sized (0.3–0.7) μm × (1.0–2.2) μm. A single polar flagellum was revealed on the electronic micrographs of negatively stained strains, and the mol% G+C of the DNA was 50.3. The molecular identification of the 16S rRNA gene identified the strains as genus Listonella . The sequenced 16S rRNA gene of strain S010610-1 (GenBank accession No. AY963630 ) is 1466bp in length, that of strain S010610-3 (No. AY963631) is 1416bp, and that of strain S010623-1 (No. AY963632) is 1424bp. They shared high similarity at 97%–99% to that of Listonella .
Artificial infected flounder and turbot showed apparently identical syndrome to that of natural ones. Smears of the liver from artificially infected dead fish were Gram-stained, showing many Gram-negative rods.
The isolates are L. anguillarum of Listonella, indicated in morphology, physiology and bioinformics, and partial 16S rRNA gene sequencing. |
| Key words: Turbot Scophthalmus maximus, Listonella anguillarum, Identification, Phylogenetic analysis |
