摘要
2020年5月,广东广州某野生动物保护基地爆发疑似病毒引发的疾病。现场采样发现,病鲵体长约2 m,反应迟钝,体表有出血点或溃疡症状。本研究采用胖头鲤肌肉细胞系(fat head minnow epithelial cells,FHM)培养、超薄切片透射电镜观察、病毒主要衣壳蛋白(major capsid protein,MCP)克隆与测序分析等方法,从患病大鲵中分离得到一株病毒,鉴定其属于虹彩病毒科蛙病毒属,命名为大鲵虹彩病毒广州分离株(CGSIV-GZ)。FHM经患病大鲵组织匀浆液接种后出现细胞圆缩、死亡、脱落等典型的细胞病变症状。将感染后的FHM细胞制作超薄切片,通过电镜观察发现,FHM细胞中存在大量直径约100~120 nm具囊膜的正六边形成熟病毒粒子,形态与虹彩病毒相似。根据虹彩病毒MCP基因保守区域序列设计特异性引物对病鲵组织样本进行PCR扩增,获得了431 bp的目的基因片段,测序后经采用BLAST软件分析,发现其与GenBank中的蛙病毒衣壳蛋白基因同源性达96%~99%。确认本次野生动物保护基地大鲵发生大规模死亡是蛙病毒感染所致。
One suspected virus-induced disease broke out in May 2020 in a Wildlife conservation base in Guangzhou,Guangdong Province.The diseased Andrias davidianus showed a slow reaction.The body length of diseased A.davidianus samples was 2m with typical clinical signs such as hemorrhages on the skin and ulcer symptoms.Fat head minnow epithelial cells(FHM)were cultured,the ultrathin section of transmission electron microscope and major capsid protein(MCP)cloning and sequencing were used in the experiment.One strain of virus from diseased A.davidianus was isolated,identified,and named as Guangzhou isolate of A.davidianus iridovirus(named CGSIV-GZ).The tissue homogenate of diseased fish could cause typical cytopathic effect such as cell shrinkage,death,and exfoliation in FHM cells.Electron microscopy observation showed a large number of virus particles about 100~120 nm in diameter in infected FHM cells,which was similar to the iridescent virus.The conserved region of the major capsid protein(MCP)gene of iridovirus was amplified by PCR,and a 431bp specific gene fragment was obtained.Sequence alignment analysis of the DNA fragment showed that CGSIV-GZ shared a high identity(96%~99%)with the published MCP gene sequence of iridovirus.It was confirmed that the large-scale death of A.davidianus was caused by frog virus infection in the wildlife conservation base.This study will provide some reference basis for the prevention and control of disease induced by CGSIV in A.davidianus culture.
作者
喻大鹏
程俊
夏洪丽
夏立群
蔡佳
鲁义善
YU Da-peng;Cheng Jun;XIA Hong-li;XIA Li-qun;CAI Jia;LU Yi-shan(Shenzhen Research Institute of Guangdong Ocean University,Shenzhen 518108,Guangdong,China;Fisheries College of Guangdong Ocean University,Zhanjiang 524088,Guangdong,China;Guangdong Provincial Engineering Research Center for Aquatic Animal Health Assessment, Zhanjiang 524088,Guangdong,China;Guangdong Provincial Engineering Research Center for Aquatic Animal Health Assessment,Shenzhen 518108,Guangdong,China;Wuhan Airport Economic and Technological Development Zone Agricultural Development Investment Group Co.,Ltd.Wuhan 430040,Guangdong,China;Wuhan Shengfeng Agricultural Development Co.,Ltd.Wuhan 430040,Guangdong,China)
出处
《淡水渔业》
CSCD
北大核心
2022年第4期11-17,共7页
Freshwater Fisheries
基金
深圳市科技计划项目(JCYJ20180507183240459)
广西自然科学基金(2020GXNSFAA297243)
南方海洋科学与工程广东省实验室(湛江)(湛江湾实验室)自主立项项目(ZJW-2019-06)
广东海洋大学“冲一流”专项资金项目(231419013)
广西红树林滨海湿地生态保护与可持续利用人才小高地人才资助项目(BGMRC202101)。
