大口黑鲈弹状病毒的分离与鉴定  

Isolation and identification of Micropterus salmoides rhabdovirus

作　　者：章文言 张玉军 李陈 石和荣 刘学芹 ZHANG Wenyan;ZHANG Yujun;LI Chen;SHI Herong;LIU Xueqin(College of Fisheries,Huazhong Agricultural University/Hubei Engineering Technology Research Center for Aquatic Animal Diseases Control and Prevention,Wuhan 430070,China;Dongguan Yinhua Biotechnology Co.,Ltd.,Dongguan 523000,China)

机构地区：[1]华中农业大学水产学院/湖北省水生动物病害防控工程技术研究中心,武汉430070 [2]东莞市银华生物科技有限公司,东莞523000

出　　处：《华中农业大学学报》2022年第6期230-236,共7页Journal of Huazhong Agricultural University

基　　金：国家重点研发计划项目(2018YFD0900505)。

摘　　要：为探明广东省韶关市某大口黑鲈(Micropterus salmoides)养殖场的大口黑鲈幼鱼大量死亡的原因,对患病鱼进行临床观察,并从细菌学、寄生虫学、病毒学三方面进行检测,排除细菌和寄生虫感染的可能性后,通过RT-PCR检测、细胞分离培养、人工感染、组织病理切片苏木精-伊红(H&E)染色、超薄切片透射电镜观察、系统发育树分析,初步确定分离得到1株大口黑鲈弹状病毒(micropterus salmoides rhabdovirus,MSRV),命名为大口黑鲈弹状病毒韶关株(MSRV-SG01)。人工感染试验结果显示,试验鱼2 d内出现死亡,并伴随出血、烂尾、拖便等临床症状,7 d内死亡率达100%,通过组织病理学切片观察到病鱼的肝脏、脾脏均呈现大面积坏死,与自然患病鱼症状相符。根据G蛋白氨基酸序列,将分离到的MSRV-SG01毒株与GenBank中已报道的其他的弹状病毒进行系统发育树分析比对,结果显示,MSRV-SG01毒株与MSRV-FJ985、MSRV-YH01、SCRV聚为一类,且与已报道的MSRV-FJ985毒株、MSRV-YH01毒株同源性高达97%以上。通过以上的试验分析,确定幼鱼大量死亡的原因为MSRV感染。In order to find out the cause of the massive death of largemouth bass(Micropterus salmoides)juveniles in a breeding farm in Shaoguan City,Guangdong Province,clinical observation,and bacteriological,parasitological and virological detection of the affected fish were carried out.And after excluding the possibility of bacterial and parasitic infections,RT-PCR detection,cell isolation and culture,artificial infection,hematoxylin-eosin(H&E)staining of histopathological sections,transmission electron microscope observation of ultrathin sections and phylogenetic tree analysis were performed and a Micropterus salmoides rhabdovirus(MSRV)was isolated and named as the Micropterus salmoides rhabdovirus Shaoguan strain(MSRV-SG01).The results of the artificial infection test showed that the experimental fish died within 2 days,accompanied by clinical symptoms such as hemorrhage,rotten tail,and dragging stools,and the mortality rate reached 100%within 7 days.Histopathological sections showed that the liver and spleen of the diseased fish showed extensive necrosis,which was consistent with the symptoms of naturally diseased fish.According to the amino acid sequence of protein G,the MSRV-SG01 strain isolated in this study was compared with other rhabdoviruses reported in GenBank by phylogenetic tree analysis and the result showed that FJ985,MSRV-YH01,SCRV,and SCRV are grouped together,and the similarity with the reported MSRV-FJ985 and MSRV-YH01 strains were higher than 97%.Through the above experimental analysis,it was determined that the cause of mass death of largemouth bass juveniles was MSRV infection.

关 键 词：大口黑鲈 弹状病毒 病毒分离 病毒鉴定 源头防控 水生动物疾病 

分 类 号：S852.659.5[农业科学—基础兽医学] Q959.483[农业科学—兽医学]