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首页> 中文期刊> 《水产学报》 >杂交鳢(斑鳢♀×乌鳢♂)内脏类结节病病原菌的分离、鉴定与特性分析

杂交鳢(斑鳢♀×乌鳢♂)内脏类结节病病原菌的分离、鉴定与特性分析

         
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Hybrid snakehead (Channa maculata 9 x C. argus S) is one of the most popular species cultured in the areas of eastern and southern China. With high density culture and rapid expanding production, problems of its diseases become more and more serious. A disease with white spots in internal organs of hybrid snakehead broke out in Fushan and Guangzhou in Guangdong Province in 2011. Bacteria pathogens were isolated from diseased hybrid snakehead cultured in pond. Two bacteria strains with strong virulence were chosen for identifications. The morphological, biochemical and physiological characteristics of the two isolates were studied by using conventional method, ATB Expression System(ID32 STREP)and other assays, and the characteristics of the isolated strains were in accordance with those of Aeromonas schuber-tii. Molecular analyses of 16S rRNA and gyrB genes were used for further identification. BLAST showed that 16S rRNA and gyrB genes of two strains possessed high similarities with their counterparts registered in GenBank and with each other. The virulent tests were carried out by using the pure culture of the isolated strains artificially infected hybrid snakehead, and the result indicated that the high concentration of bacteria (≥1 ×107 CFU/mL) caused acute death, and infected diseased fish displayed symptom of sarcoi-dosis after three days, while the control group showed no signs in 7d post-challenge. The morphological and biochemical characteristics of the re-isolated bacteria from the artificially infected diseased fish were the same as those of the original infected bacteria. Antibiotic sensitivity assays showed that among 16 antibiotics tested 14 were sensitive,and 2 were resistant.%从广东佛山、广州两地养殖场患内脏类结节病杂交鳢(Channa maculata♀ × C.argus ♂)内脏器官分离到2株细菌,纯化培养后获得2个分离株,编号为WL-1和WL-2,对分离菌株进行了细菌鉴定、致病性分析及药敏实验.应用常规生理生化鉴定和ATB系统细菌自动鉴定仪对分离菌株进行细胞形态学、理化特性分析,初步判定所分离菌为舒伯特气单胞菌.采用16S rRNA基因、DNA促旋酶的B亚单位蛋白(gyrB)基因对分离菌株进行DNA分子鉴定,结果显示,两个菌株间的16S rRNA基因序列相同,gyrB基因序列同源性为99.7％;分离菌株与GenBank上登录的舒伯特气单胞菌16S rRNA基因序列和gyrB基因序列同源性均最高,达99％以上;分离菌株在系统进化树上与舒伯特气单胞菌聚为一族,进一步确认分离株为舒伯特气单胞菌.人工感染健康鱼后出现与自然发病相似的内脏类结节病症状,从发病鱼内脏组织再分离的细菌特性与原感染菌相同.综合理化特性分析、基因鉴定和人工感染实验确认舒伯特气单胞菌是杂交鳢内脏类结节病的致病菌.药敏实验发现分离菌株对头孢唑啉、庆大霉素等14种药物敏感;对青霉素G、苯唑西林2种药物耐受.

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