泥蚶致病性哈维氏弧菌的分离与鉴定  

Isolation and Identification of Pathogenetic Vibrio harveyi from Ark Shell Tegillarca granosa

作　　者：滕爽爽[1] 张炯明[1] 张利兵 谢尚庶 王瑶华[1] 肖国强[1,4] 杨千元 方军 TENG Shuangshuang;ZHANG Jiongming;ZHANG Libing;XIE Shangshu;WANG Yaohua;XIAO Guoqiang;YANG Qianyuan;FANG Jun(Wenzhou Key Laboratory of Marine Genetics and Breeding,Zhejiang Key Laboratory of Exploitation and Preservation of Coastal Bio-Resource,Zhejiang Mariculture Research Institute,Wenzhou 325005,China;Yueqing Agricultural and Rural Bureau,Wenzhou 325699,China;Ruian Agricultural and Rural Bureau,Wenzhou 325200,China;College of Fisheries and Life Science,Shanghai Ocean University,Shanghai 316022,China)

机构地区：[1]浙江省海洋水产养殖研究所,浙江省近岸水域生物资源开发与保护重点实验室,温州市海洋生物遗传育种重点实验室,浙江温州325005 [2]乐清市农业农村局,浙江温州325699 [3]瑞安市农业农村局,浙江温州325200 [4]上海海洋大学水产与生命学院,上海316022

出　　处：《水产科学》2022年第4期547-555,共9页Fisheries Science

基　　金：国家重点研发计划项目(2020YFD0900802,2018YFD0901405-2);浙江省农业(水产)新品种选育重大科技专项(2021C02069-7-02);浙江省重点研发计划项目(2019C02045);财政部和农业农村部现代农业产业技术体系项目(CARS-49);温州市农业新品种选育协作组项目(2019ZX001-1).

摘　　要：自浙江发病泥蚶中分离到1株优势菌J14,采用人工感染试验、形态及生化特征分析、16S rRNA基因序列测定和药敏试验等方法,对该菌株进行鉴定并分析其相关特性。结果表明,菌株J14感染泥蚶的96h半致死密度为7.74×10^(6)个/mL,对泥蚶具有较强的致病性。菌株J14为革兰氏阴性细菌,经硫代硫酸盐柠檬酸盐胆盐蔗糖琼脂培养基分离为黄色不发光菌落,结合生理生化特征和16S rRNA基因序列分析,鉴定该菌株为哈维氏弧菌。毒力基因的携带情况显示,哈维氏弧菌J14可检测到群体效应调节基因luxR,毒力调控基因toxR,溶血素基因vhhA、vhhB,毒力相关基因toxS,几丁质酶基因chiA,丝氨酸蛋白酶基因7种毒力基因。药敏结果显示,哈维氏弧菌J14对青霉素、氨苄西林、阿莫西林、杆菌肽4种药物不敏感,对红霉素、多黏菌素B中度敏感,对庆大霉素、卡那霉素、复方新诺明等26种常用抗生素药物敏感。研究结果可为泥蚶弧菌病的防治提供参考资料。In order to study the species and related characteristics of the pathogenic strain J14 which was isolated from the sick ark shell Tegillarca granosa in Zhejiang province,the infection test,morphological observation,physiological and biochemical test,and 16S rRNA gene sequence analysis were performed.The results showed that the J14 strain was strong pathogenicty as the LC_(50) of J14 to ark shell was 7.74×10^(6)cells/mL.The strain of J14 was gram negative,and exhibited round,bulged and yellow in thiosulfate-citrate-bile-salts-sucrose agar.Biochemical characteristics showed that the strain belonged to the Vibrio.The sequence analysis of 16S rRNA gene revealed that the strain shared high homology with V.harveyi using BLAST from GenBank database and the strain clustered with V.harveyi into one branch in the phylogenetic tree analysis.According to the morphological and physiological-biochemical characteristics,as well as the results of 16S rRNA gene sequence analysis,the strain was identified as V.harveyi.The results of virulence gene test showed that the strain carried seven typical virulence genes,including luxR,toxR,vhhA,vhhB,toxS,chiA and serine protease genes.The susceptibility test showed that the strain was not sensitive to penicillin,ampicillin,amoxicillin and bacitracin,moderately sensitive to erythromycin,polymyxin B,and sensitive to 26 different antibiotics such as gentamicin,kanamycin,and cotrimoxazole.The findings provide a reference with the identification of V.harveyi and the prevention and control of bacterial disease in ark shell.

关 键 词：泥蚶 哈维氏弧菌 16SRRNA基因 毒力基因 药敏试验 

分 类 号：S917.1[农业科学—水产科学]