条石鲷检出的虹彩病毒特性研究  

Characterization of an Iridovirus Detected in Rock Bream(Oplegnathus fasciatus;Temminck and Schlegel)

作　　者：李华[1] 孙志鹏[1] 李强[1] 江育林[2] 

机构地区：[1]大连海洋大学农业部海洋水产增养殖学重点实验室,大连116023 [2]中国检验检疫科学研究院动植物检疫研究所水生动物疾病研究中心,北京100029

出　　处：《病毒学报》2011年第2期158-164,共7页Chinese Journal of Virology

基　　金：海洋公益性行业科研专项(200905020);辽宁省海洋与渔业厅项目(201005)资助

摘　　要：2009年8月份辽宁地区某网箱养殖场出现条石鲷大量死亡病情,对病鱼组织切片观察,在脾脏和肾脏等器官内发现许多形态异常肿大的嗜碱性粒细胞。病鱼组织匀浆液感染GF细胞出现了明显的细胞病变效应（CPE）。经透射电镜观察发现,脾脏、肾脏、肝脏及肠组织细胞质内存在大量呈六边形的病毒粒子,该病毒由核衣壳（100110nm）和囊膜构成,直径150180nm,似虹彩病毒。使用真鲷虹彩病毒（RSIV）959bp PstI片段特异性引物对病鱼各脏器组织进行PCR扩增,在脾脏、肾脏、鳃、肠、心脏和脑扩增出570bp大小的目的片段。同时,针对虹彩病毒主衣壳蛋白序列进行PCR扩增后得到1 400 bp大小的基因片段。病毒主衣壳蛋白序列系统进化分析显示,该病毒与真鲷虹彩病毒（RSIV-U1）等几株病毒位于同一进化分枝内。根据上述实验结果,作者认为引发条石鲷大批死亡的病原为真鲷虹彩病毒（Red sea bream iridovirus,RSIV）。During the summer of 2009,mass mortality was observed in cage-cultured Rock Bream（Oplegnathus fasciatus;Temminck and Schlegel） in the Liaoning Province.Histopathogic studies of the affected fish showed enlarged basophilic cells in the kidney and spleen.These necrotic cells were stained purple using haematoxylin and eosin（HE）.GF cell cultures showed advanced cytopathic effects after infection with virus supernatants from diseased fish homogenate.Transmission electron microscopy revealed hexagonal outlines virions in the cytoplasm of the spleen,kidney,liver,intestine cells.The viral particles consisted of a central nucleocapsid（100-110 nm） and envelope,and were 150-180 nm in diameter.These results suggested that the virus belonged to the Iridoviridae.Using polymerase chain reaction（PCR）,approximately 570-bp fragments were amplified from the viral DNA in spleen,kidney,gill,intestine,heart and brain of diseased fish with the primers derived from red sea bream Iridovirus（RSIV）.In addition,a specific fragment of 1 400 bp of the major capsid protein（MCP） gene of the Iridovirus was amplified by PCR.A phy1ogenetic tree was constructed to compare the corresponding genetic sequences in Megalocytivirus.The tree demonstrated that RSIV-LN09 virus existed in the same branch as the RSIV-U1 et al.Our present results indicated that RSIV was the causative agent.

关 键 词：条石鲷 虹彩病毒 肿大细胞 

分 类 号：S941.419[农业科学—水产养殖]