斜带髭鲷内脏白点病病原的分离鉴定及耐药性分析  

Isolation,identification and antibiotic sensitivity analysis of bacterial pathogen from Hapalogenys nitens with white spot disease in internal organs

作　　者：李慧耀 杨求华[1,2] 葛辉 何丽斌[1,2] 吴丽云 吴建绍[1,2] LI Huiyao;YANG Qiuhua;HE Libin;WU Liyun;WU Jianshao(Fisheries Research Institute of Fujian,Xiamen 361013,China;Key Laboratory of Cultivation and High-Value Utilization of Marine Organisms in Fujian Province,Xiamen 361013,China)

机构地区：[1]福建省水产研究所,福建厦门361013 [2]福建省海洋生物增养殖与高值化利用重点实验室,福建厦门361013

出　　处：《应用海洋学学报》2023年第4期643-655,共13页Journal of Applied Oceanography

基　　金：福建省属公益类科研院所基本科研专项(2020R1013005)。

摘　　要：本研究采用平板分离法对具有典型症状的斜带髭鲷(Hapalogenys nitens)进行病原微生物的分离培养,结合形态学、生理生化特征和分子生物学技术对病原进行种属鉴定,并对分离得到的病原菌进行了药物敏感性试验。病理切片结果显示,病鱼肝脏、脾脏、鳃、头肾、心脏均可观察到明显的白色炎性肉芽肿病灶,从患病鱼脾脏、肝脏和头肾的白色结节处均分离到单一外观的细菌菌落hn-1。通过生理生化特征鉴定、16S rRNA基因测序和回归感染实验证实斜带髭鲷内脏白点病的致病菌为哈维氏弧菌(Vibrio harveyi)。透射电镜观察显示分离菌具有极生鞭毛及侧鞭毛;毒力基因检测显示分离菌携带toxR、luxR、vvh、vhh和trh基因;药敏试验结果显示,分离菌株主要对喹诺酮类、氨基糖苷类、四环素类等药物具有较强的敏感性。研究结果可为斜带髭鲷的内脏白点病的病害防控提供科学依据。The purpose of this study was to identify the pathogen of white spot disease of Hapalogenys nitens and analyze its antibiotic sensitivity,which could provide scientific basis for diagnosis and prevention of the disease.The pathogenic microorganisms of H.nitens with typical symptoms were isolated by plate separation method.Pathogenic bacteria are identified with morphological,physiological and biochemical characteristics and molecular sequencing techniques.Furthermore,the drug sensitivity test was carried out on the isolated pathogenic bacteria.The pathological section showed that white inflammatory granuloma lesions were observed in the liver,spleen,gills,head kidney and heart of the fish infected.Single colony named hn-1 can be isolated from the spleen,liver and head kidney of the infected and identification of physiological and biochemical characteristics,gene sequencing and regression infection experiments confirmed that Vibrio harveyi was the pathogenic bacterium of visceral white spot disease of the fish infected.Under the electron microscope,the isolated bacterium was observed to have polar flagella and lateral flagella.The virulence gene test showed that the isolates carried a variety of virulence genes,including toxR,luxR,vvh,vhh and trh.Results of drug sensitivity test showed that the isolated strains were mainly sensitive to quinolones,aminoglycosides,tetracyclines and other drugs.The results can provide scientific basis for the prevention and control of visceral white spot disease of H.nitens.

关 键 词：海洋生物学 水产养殖 斜带髭鲷 内脏白点病 哈维氏弧菌 耐药性分析 

分 类 号：P735[天文地球—海洋生物学]