南宁市售文蛤中弧菌的分离鉴定

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【目的】对从南宁市售文蛤体内分离获得的待测菌株进行鉴定,为掌握了解广西文蛤产品中弧菌科细菌的带菌情况提供参考依据。【方法】用硫代硫酸盐柠檬酸盐胆盐蔗糖琼脂培养基(TCBS)从南宁海鲜市场的文蛤中分离获得4株细菌,观察其菌落形态,对分离菌株进行生理生化特性鉴定及16SrRNA基因序列进行克隆、测序,运用DNASTAR中的MegAlign软件进行序列分析,并构建系统进化树。【结果】TCBS培养基30℃培养24h后,菌株No.8和No.9生长良好,形成直径2mm左右、圆形、隆起的菌落;菌株No.10、No.11生长不良,部分形成直径1mm左右、圆形、隆起的黄色菌落。菌株生化鉴定结果表明,菌株No.8、No.9在无盐蛋白胨水中不生长,且不能分解葡萄糖;而菌株No.10、No.11可在无盐蛋白胨水中生长,且能分解葡萄糖并产气。16SrRNA序列系统进化树显示,菌株No.8与副溶血弧菌聚集为一个分支,No.9与河流弧菌聚集为一个分支,No.10、No.11则分别与斑点气单胞菌、嗜水气单胞菌聚集为一个分支。综合表型和分子特征可知,菌株No.8为副溶血弧菌,No.9为河流弧菌,No.10为斑点气单胞菌、No.11为嗜水气单胞菌。细菌药敏结果显示,4株分离菌株对四环素、环丙沙星、先锋噻肟、先锋必均表现为高度敏感。【结论】南宁市售文蛤产品主要是污染了弧菌属及气单胞菌属细菌,生产上可选用四环素、环丙沙星、先锋噻肟、先锋必等药物进行防治。 【Objective】 The objective of this study was to identify the strains isolated from the cultured clams in Nanning City and to provide a reference for understanding the bacteriolytic activity of the bacteria in the clams of Guangxi clams. 【Method】 Four strains of bacteria were isolated from the clam in Nanning seafood market by using the thiosulfate citrate bile salt sucrose agar medium (TCBS). The colony morphology was observed. The physiological and biochemical characteristics and 16S rRNA gene sequence of the isolates were identified Cloned, sequenced and sequenced using MegAlign software in DNASTAR. The phylogenetic tree was constructed. 【Result】 After cultured at 30 ℃ for 24 h in TCBS medium, strain No.8 and No.9 grew well, forming round and bulged colonies with diameter of about 2mm. Strain No.10 and No.11 were poorly grown and partially formed with diameter of 1mm Around, round, raised yellow colonies. The results of biochemical identification showed that strain No.8 and No.9 did not grow in salt-free peptone water, and could not decompose glucose. Strain No.10 and No.11 could grow in salt-free peptone water and decompose glucose gas. 16S rRNA sequence phylogenetic tree showed that strain No. 8 and V. parahaemolyticus gathered as a branch, No. 9 and V. angustida gathered as a branch, No. 10 and No. 11, respectively, with Aeromonas cavaci, Aeromonas aggregates as a branch. According to the comprehensive phenotype and molecular characteristics, Vibrio parahaemolyticus strain No. 8, Vibrio river Vibrio strain No. 9, Aeromonas cavacii strain No. 10, and Aeromonas hydrophila strain No. 11. Bacterial susceptibility results showed that four isolates were highly sensitive to tetracycline, ciprofloxacin, avant-garde, and pioneer. 【Conclusion】 The main clam products in Nanning are contaminated with Vibrio and Aeromonas. The production of Tetracycline, Ciprofloxacin, Pioneer, and Pioneer can be controlled by the drug.
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