一株似鲇高原鳅源蛙病毒的分离与鉴定
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四川农业大学 动物医学院, 四川 雅安 625014

作者简介:

范玉蕾(1990–), 女, 研究生, 研究方向水生动物疾病学. E-mail: 449587713@qq.com

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S941

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四川省科技支撑计划项目(2014NZ0027).


Isolation and identification of a ranavirus from Triplophysa siluroides
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College of Veterinary Medicine, Sichuan Agricultural University, Ya’an 625014, China

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    摘要:

    从四川乐山某养殖场自然发病的似鲇高原鳅(Triplophysa siluroides)体内分离到一株病毒 FYL140220。病毒 感染鲤上皮瘤细胞系(epithelima popuasum cuprini, EPC)后, 细胞呈现圆缩、坏死、脱落、明显空斑的病变特征。将 自然发病鱼组织过滤除菌液和细胞培养病毒液分别接种健康似鲇高原鳅, 试验鱼出现与自然发病鱼相同的症状, 死亡率分别为 30%和 40%, 而对照组无异常。对经 FYL140220 感染出现典型细胞病变(CPE)的 EPC 细胞制备超薄 切片进行电镜观察, 发现病毒呈正六边形, 对称 20 面体, 对角线直径约(103±7) nm; 提取细胞培养病毒液、自然 发病鱼和人工感染发病鱼的内脏器官总 DNA 进行蛙病毒主要衣壳蛋白(maior capsid protein, MCP)基因保守区域 的 PCR 扩增, 均能扩增出预期大小约 500 bp 的特异性条带。MCP 基因同源性与遗传进化分析表明, 分离株 FYL140220 与蛙病毒属成员聚为一支, 尤其与大鲵蛙病毒与沼泽绿蛙病毒同源性最高, 分别达 99.8%和 99.6%。结 合 PCR 检测、电镜观察和系统发育分析, 确认分离病毒 FYL140220 为蛙病毒, 这是首次报道蛙病毒可自然感染 似鲇高原鳅并致死。

    Abstract:

    A virus named FYL140220 was isolated from naturally infected Triplophysa siluroides in Leshan, Sichuan Province using epithelima popuasum cuprini (EPC) cells. The infected EPC cells showed circular shrinkage, necrosis, and desquamation, which formed significant plaque-lesion characteristics. Diseased tissue suspension filtered from bac- teria and EPC-grown virus were used to inoculate healthy Triplophysa siluroides. As a result, the infected T. siluroides developed similar clinical symptom to those described above and suffered 30% and 40% mortality, whereas the uninfected control EPC cells remained normal. Electron microscopy revealed icosahedral viral particles in the cells. The FYL140220 virus had an average diameter of (103 ± 7) nm, and the hexagonal shape was highly similar to that of other Iridoviridae viruses. DNA was extracted from the EPC-grown virus and naturally and artificially infected internal T. siluroides tissues for polymerase chain reaction (PCR) amplification of the conserved region of the ranavirus major capsid protein (MCP) gene, revealing a 500-bp fragment. The MCP homology and genetic evolution analysis showed that FYL140220 and other ranavirus strains formed a tight cluster and shared 99.8% identity with CGSV-G and 99.6% with RGV. Taken together, these results confirm that FYL140220 is a ranavirus. This is the first report on a natural ranavirus infection and mortality caused by this pathogen in cultured Triplophysa.

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范玉蕾,耿毅,周燕,邓梦玲,余泽辉,汪开毓,黄小丽,陈德芳,张雨薇.一株似鲇高原鳅源蛙病毒的分离与鉴定[J].中国水产科学,2015,22(3):556-562
FAN Yulei, GENG Yi, ZHOU Yan, DENG Mengling, YU Zehui, WANG Kaiyu, HUANG Xiaoli, CHEN Defang, ZHANG Yuwei. Isolation and identification of a ranavirus from Triplophysa siluroides[J]. Journal of Fishery Sciences of China,2015,22(3):556-562

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  • 在线发布日期: 2015-05-13
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