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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(100~110nm)和囊膜构成,直径150~180nm,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)。
In August 2009, a large number of dead cichlids died in a cage farm in Liaoning Province. Tissue slices were observed for diseased fish and many abnormally enlarged basophils were found in spleen and kidney. Disease fish tissue homogenate infected GF cells showed a significant cytopathic effect (CPE). The results of transmission electron microscopy showed that a large number of hexagonal virus particles existed in the cytoplasm of spleen, kidney, liver and intestine. The virus consisted of nucleocapsid (100-110 nm) and envelope, with a diameter of 150-180 nm. . Using the 959bp PstI fragment-specific primer of iris virus (RSIV), the tissues of diseased fish were amplified by PCR and the target fragment of 570bp was amplified in spleen, kidney, gill, intestine, heart and brain. At the same time, a gene fragment of 1 400 bp was obtained after PCR amplification of the main capsid protein sequence of iridescent virus. Phylogenetic analysis of the main capsid protein sequence of the virus showed that the virus was located in the same evolutionary branch with several viruses such as the true bream iridescent virus (RSIV-U1). Based on the above experimental results, the authors believe that the cause of the large number of deaths of the bar snub-nosed sea bream iridovirus (Red sea bream iridovirus, RSIV).