1株乌鳢水泡病毒的分离与鉴定  

Isolation and Identification of a Strain of Snakehead Fish Vesiculovirus

作　　者：李园园[1] 袁倩 冯晓然 邓思 李娜 姚伦广[1,2,3,4] 秦英惠 LI Yuanyuan;YUAN Qian;FENG Xiaoran;DENG Si;LI Na;YAO Lunguang;QIN Yinghui(College of Life Science,Nanyang Normal University,Nanyang 473061,China;Henan Field Observation and Research Station of Headwork Wetland Ecosystem of the Central Route of South-to-North Water Diversion Project,Nanyang 473061,China;Henan Provincial Engineering and Technology Center of Health Products for Livestock and Poultry,Nanyang 473061,China;Key Laboratory of Ecological Security and Collaborative Innovation Centre of Water Security for Water Source Region of Mid-line of South-to-North Diversion Project,Nanyang 473061,China)

机构地区：[1]南阳师范学院生命科学学院,南阳473061 [2]河南省南水北调中线渠首湿地生态系统野外科学观测研究站,南阳473061 [3]河南省畜禽保健品工程技术研究中心,南阳473061 [4]南水北调中线水源区生态安全重点实验室与水安全协同创新中心,南阳473061

出　　处：《中国畜牧兽医》2025年第2期811-822,共12页China Animal Husbandry & Veterinary Medicine

基　　金：河南省科技攻关项目(222102110079);中原科技创新领军人才项目(244200510040);南阳师范学院博士专项项目(2022ZX030);南阳师范学院国家自然科学基金培育项目(2023PY018)。

摘　　要：[目的]明确2021年9月河南省南阳市某乌鳢养殖场乌鳢大量死亡的病原。[方法]本研究通过体表检测、显微观察、细菌分离、PCR检测、序列相似性和系统进化树分析、细胞培养、透射电镜观察、人工感染及病理切片观察等方法进行病原体鉴定。[结果]患病鱼体表无明显机械损伤和出血点,未在患病鱼体表和鳃表面观察到寄生虫的存在,未从患病鱼的组织中分离出致病菌。PCR检测结果显示,患病鱼呈乌鳢水泡病毒(SHVV)阳性。SHVV G基因序列相似性分析显示,分离株与佛山分离株HSHRV-MS2021相似性最高,达99.85%。系统进化树显示,分离毒株与SHVV-2019、SHVV-YL01和HSHRV-MS2021株等聚为同一分支,属于Siniperhavirus属,与HSHRV-MS2021株亲缘关系最近。患病鱼组织匀浆液接种鲤鱼上皮瘤细胞(epithelioma papillosum cyprini cells, EPC)和石斑鱼脾脏细胞(grouper spleen cells, GS)均可引起明显的细胞病变。透射电镜观察发现,病变的EPC细胞内存在大量的子弹状病毒粒子。人工感染试验表明,感染鱼出现与自然患病鱼相似症状,死亡率高达100%。组织切片观察显示,回感患病鱼肠道微绒毛消失,肝脏和脾脏血淋巴增多。[结论]本研究从广东以外地区养殖的乌鳢中分离出1株SHVV,命名为SHVV-NY01,该养殖场乌鳢的大量死亡是由SHVV引起的,本研究结果为该病毒的流行病学调查和防控提供了参考。[Objective]This study was aimed to clarify the pathogen,which caused mass mortality of snakehead fish in Nanyang city,Henan province,in September 2021.[Method]This study identified pathogens through methods such as surface detection,microscopic observation,bacterial isolation,PCR detection,phylogenetic tree analysis,cell culture,transmission electron microscopy observation,artificial infection,and pathological section observation.[Result]No noticeable lesions and blooding points was observed on the surface of diseased fish.No parasite in the gills or on the body surfaces was found,and no pathogenic bacterium was isolated.PCR detection result showed that the diseased fish was positive for Snakehead fish vesiculovirus(SHVV).The similarity analysis of SHVV G gene sequence showed that the isolate had the highest similarity with the Foshan isolate HSHRV-MS2021,reaching 99.85%.The phylogenetic tree showed that the isolated strain was clustered into the same branch as SHVV-2019,SHVV-YL01 and HSHRV-MS2021 strains,belonging to the Siniperhavirus genus,and was most closely related to the HSHRV-MS2021 strain.The tissue homogenate of diseased fish could cause obvious cytopathic effect in EPC and GS cells.Transmission electron microscopy observations showed that abundant bullet shaped virus particles scattered on the surface of diseased EPC cells.Artificial infection experiments showed that infected fish exhibited symptoms similar to those of naturally diseased fish,with a mortality rate as high as 100%.Observation of tissue slices showed that the microvilli in the intestinal tract of the fish with recurrent infection disappeared,and the hemolymph in the liver and spleen increased.[Conclusion]In this study,a strain of SHVV was isolated from snakehead fish cultured outside Guangdong province and named SHVV-NY01.A large number of deaths of snakehead fish in this farm were caused by SHVV.The results of this study provided a reference for the epidemiological investigation and control of this virus.

关 键 词：乌鳢 弹状病毒 乌鳢水泡病毒 分离 鉴定 

分 类 号：S852.65[农业科学—基础兽医学]