Establishment and susceptibility to cyprinid herpesvirus 2 of Carassius carassius gill cell line

Abstract

Haematopoietic necrosis in Carassius auratus is one of the most serious diseases in recent years caused by CyHV-2 (cyprinid herpesvirus 2) which has caused huge economic losses in aquaculture. Currently, the isolation and propagation of CyHV-2 in vitro is difficult due to the lack of sensitive cell lines, which hinder further studies on the pathogenesis and gene functions of the virus. In this study, Carassius carassius gill cell line (CCG) was constructed and had been passaged up to 90 passages at 25 ℃ in Medium199 (M199) with 15–20% Foetal bovine serum (FBS). Cellular karyotype analysis showed that the chromosome number was 46 at passage 50. CCG could be successfully transfected with GFP reporter gene with an efficiency of more than 60% suggesting that CCG could be used for exogenous gene expression in vitro. CCG infected with CyHV-2 displayed typical cytopathic effects (CPE), and viral replication was confirmed by PCR, indirect immunofluorescence assay (IFA) and transmission electron microscopy (TEM) assay, respectively. In conclusion, the CCG cell line can be used as a useful tool to study the propagation and pathogenesis of CyHV-2.

Keywords

• Carassius carassius
• Cell line
• CyHV-2
• Cytopathic effect