Establishing the optimal fetal bovine serum concentration to support replication of cyprinid herpesvirus 3 in CCB and KF-1 cell lines
2020
Kim, Hyoung Jun | Kwon, Se Ryun | Yuasa, Kei
Koi herpesvirus (KHV) disease is a serious disease in cultured carp (Cyprinus carpio). CCB and KF-1 cell lines are commonly used for virus isolation and observation of cytopathic effects (CPE) in carp and koi samples. The purpose of this study was to determine the optimal concentration of fetal bovine-serum (FBS) to use for supporting the replication of cyprinid herpesvirus 3 CyHV-3 in CCB and KF-1 cell lines. The following concentrations were tested: 0%, 2%, 5%, and 10% FBS. At 7 days post-viral inoculation (dpi), CPE with clear vacuolation was observed in both cell lines when supplemented with 0 and 2% FBS, but not in those supplemented with 5% or 10% FBS. At 14 dpi, CPE was observed in both cell lines supplemented with FBS at any of the tested concentrations when a high virus titer was inoculated. However, CPE was indistinguishable between cell lines supplemented with 10% FBS when a low virus titer was inoculated. Results of qPCR indicated that the number copies of the viral genome tended to be larger in both cell lines supplemented with 10% FBS than the corresponding number in cell lines supplemented with 0%, 2%, or 5% FBS, at 7 dpi. In conclusion, we recommend using 2% FBS as supplement for isolation and diagnosis of CyHV-3 viral infection in carp samples.
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