养殖三疣梭子蟹体内哈维氏弧菌的分离与致病性鉴定  

Isolation and identification of pathogenic Vibrio harveyi from swimming crab,Portunus trituberculatus

作　　者：周永强[1] 李登峰[1] 彭姣[1] Ester Johanna Shikongo 陈炯[1] 刘联国[1] 顾晓英[1] 

机构地区：[1]宁波大学海洋学院,浙江宁波315211

出　　处：《应用海洋学学报》2013年第2期215-221,共7页Journal of Applied Oceanography

基　　金：宁波市农业攻关课题资助项目(2011C10004);宁波大学人才工程资助项目(XR0713017);宁波市海洋蟹类产业科技创新团队资助项目(2011B81003);宁波大学王宽诚幸福基金资助项目

摘　　要：养殖的三疣梭子蟹(Portunus trituberculatus)秋、冬季常发生非典型症状的死亡.本研究取濒死的三疣梭子蟹内脏及体腔液进行细菌分离,经纯化传代获得1株优势菌HY.HY菌的革兰氏染色呈阴性,菌体呈杆状、弧形或逗点状,无荚膜,无芽孢,极生单鞭毛,运动性强.生长特性和生化试验表明该菌生长需盐,最适生长的NaCl含量为30 mg/cm3;28℃下生长旺盛,能利用葡萄糖产酸但不产气,可以利用麦芽糖、蔗糖等糖类.测定了HY菌约930bp的16S rDNA序列,分类进化树显示其与哈维氏弧菌(Vibrio harveyi)XC08001聚为一簇.综合形态、生化和16S rDNA序列分析结果确定HY菌为哈维氏弧菌.以2×106cfu/只(蟹)的HY菌悬液注射健康梭子蟹成蟹(平均体重为400±25 g/只),致死率为35%;菌液浸泡感染梭子蟹仔蟹的半数致死剂量(LC50)为8.25×105cfu/cm3.上述结果表明分离的哈维氏弧菌对三疣梭子蟹具有致病性.Mass mortality of farmed swimming crab, Portunus trituberculatus, with no typical symptoms are observed frequently in autumn and winter these years. A predominant bacterium HY was isolated from the viscera and coelomic fluid of infected crabs. HY proliferated well in beef agar supplied with 30 mg/cm3 NaC1 at 28℃. Yet it could not be cultured in salt free medium. HY could use glucose, maltose, sucrose and arabinose. 16S rDNA of HY was amplified, sequenced and analyzed. Plylogenetic tree based on 16S rDNA showed that HY is nearest to Vibrio harveyi XC08001. Morphological, cultural and physiological biochemical characters also demonstrated HY as Vibrio harveyi. Death rate of adult P. strituberculatu experimentally infected by injection with V. harveyi HY （ 2 10^6 cfu per crab） was up to 35%. The crab larvae were susceptible to V. harveyi infection by bath inoculation resulted in LCso of 8.25 x 10^5 cfu/cm3. Thus. it confirmed that V. harvei HY is pathogenic to P. strituberculatu.

关 键 词：海洋微生物学 三疣梭子蟹 病原 哈维氏弧菌 分离 鉴定 

分 类 号：S941.4[农业科学—水产养殖]