Abstract
Grass carp reovirus (GCRV) is a causative agent of haemorrhagic disease in grass carp that drastically affects grass carp aquaculture. Here we report a novel GCRV isolate isolated from sick grass carp that induces obvious cytopathic effect in CIK cells and name it as GCRV096. A large number of GCRV 096 viral particles were found in the infected CIK cells by electron microscope. The shape, size and the arrangement of this virus were similar to those of grass carp reovirus. With the primers designed according to GCRV 873 genome sequences, specific bands were amplified from sick grass carp and the infected CIK cells. The homology rates among vp4, vp6 and vp7 gene in GCRV 096 and those of some GCRV isolates were over 89%. In this study, the sequences of vp4, vp6 and vp7 were used to analyse sequence variation, phylogenetic relationships and genotypes in twenty five GCRV isolates. The results indicated these twenty five GCRV isolates should be attributed to four genotypes. And there were no obvious characteristics in the geographical distribution of GCRV genotype. The study should provide the exact foundation for developing more effective prevention strategies of grass carp haemorrhagic disease.
Highlights
Grass carp reovirus (GCRV), which is known as a member of the Aquareovirus genus and the Reoviridae family, can cause serious haemorrhagic disease in grass carp (Chen and Jiang 1983) and obvious cytopathic effect (CPE) on many cell lines from fish (Zuo et al 1986; Lu et al 1990)
Considering vp4, vp6 and vp7 gene as molecular makers, we investigated sequence variation characteristics, the phylogenetic relationships and genotypes of twenty five GCRV isolates to find the evolutive characteristic of GCRV in the study
Virus infection in sensitive cellls and particle identification Three days after the culture of the CIK cells infected by GCRV 096, CPE phenomenon was observed and the shedding and apoptosis occurred in most of the CIK cells five days after the infection
Summary
Grass carp reovirus (GCRV), which is known as a member of the Aquareovirus genus and the Reoviridae family, can cause serious haemorrhagic disease in grass carp (Chen and Jiang 1983) and obvious cytopathic effect (CPE) on many cell lines from fish (Zuo et al 1986; Lu et al 1990). A number of various GCRV isolates have been isolated from diseased grass carp around the world, including GCRV 873, GCRV 875, GCRV HZ08, GCRV GD108, AGCRV and others (Fang et al 2002; Chi et al 2011; Ye et al 2012; Zeng et al 2013). These isolates are distinct in their levels of virulence and cell culture characteristics, and in their antigenicity (Fang et al 2002; Mohd Jaafar et al 2008; Zhang et al 2010a). GCRV is a double-stranded RNA virus that is assigned to the Aquareovirus C species. One of the virus genotyping methods is based on the analysis of the nucleotide sequence
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