| 石斑鱼及暂养环境中副溶血弧菌分离鉴定与毒力评价 |
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| 引用本文： | 高 磊,邓益琴,刘助红,龙云映,陈 偿.石斑鱼及暂养环境中副溶血弧菌分离鉴定与毒力评价[J].海洋与湖沼,2014,45(6):1280-1287. |
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| 作者姓名： | 高 磊 邓益琴 刘助红 龙云映 陈 偿 |
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| 作者单位： | 中国科学院南海海洋研究所 中国科学院海洋生物资源可持续利用重点实验室 广东省应用海洋生物学重点实验室 广州 510301;中国科学院南海海洋研究所 中国科学院海洋生物资源可持续利用重点实验室 广东省应用海洋生物学重点实验室 广州 510301;中国科学院南海海洋研究所 中国科学院海洋生物资源可持续利用重点实验室 广东省应用海洋生物学重点实验室 广州 510301;中国科学院南海海洋研究所 中国科学院海洋生物资源可持续利用重点实验室 广东省应用海洋生物学重点实验室 广州 510301;中国科学院南海海洋研究所 中国科学院海洋生物资源可持续利用重点实验室 广东省应用海洋生物学重点实验室 广州 510301 |
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| 基金项目： | 中国科学院知识创新工程重点项目, KZCX2-EW-Q215 号; 国家自然科学基金面上项目, 31272697 号 |
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| 摘 要： | 副溶血弧菌(Vibrio parahaemolyticus,VP)存在于海洋环境和海产品中,可引起人的食物中毒。本研究的目的是对某地石斑鱼暂养环境的副溶血弧菌的致病风险进行评价。结合弧菌选择性培养基TCBS、gyr B基因和16Sr RNA特定片段的序列分析方法对分离菌株进行鉴定;通过PCR检测毒力基因,血琼脂平板溶血实验、细胞感染实验、KM鼠灌胃实验分别检测各菌株的致病性、溶血性、细胞毒性与肠毒性。共鉴定出4株具有毒力的副溶血弧菌,分别为HS51-5、HS54-4、MJ01-1和MJ03-1。所有菌株都检测到溶血素基因(tdh和tlh)、三型分泌系统(T3SS)基因簇1(T3SS-1)的4个效应蛋白基因(vop Q、vop R、vop S和vpa0450)、和基因簇2(T3SS-2)的1个效应蛋白基因(Vop A)的分布,同时在HS51-5和HS54-4还检测到T3SS-2中的其他3个效应蛋白基因(vop C、vop T和vop L)。所有菌株都具有溶血现象、细胞毒性以及对小鼠的致死能力,而HS54-4、MJ01-1、MJ03-1对小鼠的毒力明显强于HS51-5。因此,从毒力基因检测、神奈川溶血、细胞毒性和对小鼠的毒力等4个方面的结果都表明副溶血弧菌具有致病性,说明石斑鱼暂养环境的副溶血弧菌具有致病风险,应加强对这些环境的监测。
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| 关 键 词： | 副溶血弧菌 海产品 污染 毒力评价 |
| 收稿时间： | 2014/1/10 0:00:00 |
| 修稿时间： | 4/6/2014 12:00:00 AM |
VIRULENCE OF VIBRIO PARAHAEMOLYTICUS IN SEAFOOD |
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| GAO Lei,DENG Yi-Qin,LIU Zhu-Hong,LONG Yun-Ying and CHEN Chang.VIRULENCE OF VIBRIO PARAHAEMOLYTICUS IN SEAFOOD[J].Oceanologia Et Limnologia Sinica,2014,45(6):1280-1287. |
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| Authors: | GAO Lei DENG Yi-Qin LIU Zhu-Hong LONG Yun-Ying and CHEN Chang |
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| Institution: | South China Sea Institute of Oceanology, Key Laboratory of Marine Bio-resources Sustainable Utilization, CAS, Key Laboratory of Applied Marine Biology of Guangdong, Guangzhou 510301, China;South China Sea Institute of Oceanology, Key Laboratory of Marine Bio-resources Sustainable Utilization, CAS, Key Laboratory of Applied Marine Biology of Guangdong, Guangzhou 510301, China;South China Sea Institute of Oceanology, Key Laboratory of Marine Bio-resources Sustainable Utilization, CAS, Key Laboratory of Applied Marine Biology of Guangdong, Guangzhou 510301, China;South China Sea Institute of Oceanology, Key Laboratory of Marine Bio-resources Sustainable Utilization, CAS, Key Laboratory of Applied Marine Biology of Guangdong, Guangzhou 510301, China;South China Sea Institute of Oceanology, Key Laboratory of Marine Bio-resources Sustainable Utilization, CAS, Key Laboratory of Applied Marine Biology of Guangdong, Guangzhou 510301, China |
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| Abstract: | V. parahaemolyticus is an important human pathogen causing human gastroenteritis presumed by consuming uncooked seafood. However, the causation remains unknown on the incidence to seafood production and distribution. We isolated and identified pathogenic traits of V. parahaemolyticus from local seafood market and restaurants to estimate the risk and pathway of contamination. V. parahaemolyticus was isolated using TCBS medium and identified by sequencing gyrB and 16S rRNA. Distribution of virulence genes was detected by PCR. The Wagstsuma Blood Agar media, cell infection, and artificial infection of Kunming mice were used to evaluate the Kanagawa phenomenon, cytotoxicity, and pathogenicity. Four strains of V. parahaemolyticus, namely HS51-5, HS54-4, MJ01-1, and MJ03-1 were isolated, and their virulence was identified. All the strains had tdh, tlh, and T3SS-1 effectors, and vopA encoding one of the T3SS-2 effectors. None of them had trh. HS51-5 and HS54-4 had vopC, vopT, and vopL, being the genes of effectors T3SS-2. Although they were all Kanagawa positive and showed strong cytotoxicity, HS54-4, MJ01-1, and MJ03-1 were more virulent than HS51-5. The presence of virulence genes, Kanagawa phenomenon, cytotoxicity, and the induction to Kunming mice death indicated the pathogenicity of V. parahaemolyticus. Therefore, V. parahaemolyticus contamination is highly risky to fresh seafood in a temporary environment for storing and delivering, during which close monitoring to the pathogenicity is strongly recommended. |
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| Keywords: | Vibrio parahaemolyticus seafood contamination toxicity evaluation |
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