Infection and propagation of lymphocystis virus isolated from the cultured flounder Paralichthys olivaceus in grass carp cell lines
Infection and propagation of lymphocystis virus isolated from the cultured flounder Paralichthys olivaceus in grass carp cell lines
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DOI：
10.3354/dao057027
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发表时间：
2003-12-03
影响因子：
1.4
通讯作者：
Gui, HF
中科院分区：
文献类型：
--
作者：
Zhang, QY;Ruan, HM;Gui, HF
淋巴囊肿病是中国养殖牙鲆常见的淋巴囊肿病病原。在这项研究中，13个鱼类细胞系对LV的敏感性进行了测试。其中，来自淡水草鱼的2个细胞系对LV敏感，1个细胞系GCO(草鱼卵巢)因此被用来复制和繁殖病毒。细胞病变效应(CPE)在接种后1d首先在细胞单层中观察到，3d时已扩展到约75%的细胞单层。Giemsa染色和Hoechst 33258染色荧光显微镜观察细胞病变特征。用电子显微镜观察了病毒颗粒的增殖情况。超微结构分析显示了几个明显的细胞变化，如染色质紧凑和边集，小泡形成，细胞表面卷曲，核碎裂，出现特征性的‘水泡’和细胞融合。这项研究提供了LV在淡水鱼细胞系中感染和繁殖的详细报告，并提供了病毒在受感染细胞中繁殖的直接电子显微镜证据。提出了一种可能的CPE控制过程。
The causative agent of lymphocystis disease that frequently occurs in cultured flounder Paralichthys olivaceus in China is lymphocystis virus (LV). In this study, 13 fish cell lines were tested for their susceptibility to LV. Of these, 2 cell lines derived from the freshwater grass carp Ctenopharyngodon idellus proved susceptible to the LV, and 1 cell line, GCO (grass carp ovary), was therefore used to replicate and propagate the virus. An obvious cytopathic effect (CPE) was first observed in cell monolayers at 1 d post-inoculation, and at 3 d this had extended to about 75% of the cell monolayer. However, no further CPE extension was observed after 4 d. Cytopathic characteristics induced by the LV were detected by Giemsa staining and fluorescence microscopic observation with Hoechst 33258 staining. The propagated virus particles were also observed by electron microscopy. Ultrastructure analysis revealed several distinct cellular changes, such as chromatin compaction and margination, vesicle formation, cell-surface convolution, nuclear fragmentation and the occurrence of characteristic 'blebs' and cell fusion. This study provides a detailed report of LV infection and propagation in a freshwater fish cell line, and presents direct electron microscopy evidence for propagation of the virus in infected cells. A possible process by which the CPEs are controlled is suggested.