Comparative sensitivity of three new cell lines developed from gill, liver and brain tissues of goldfish, Carassius auratus (L.) to cyprinid herpesvirus-2 (CyHV-2)

Abstract

Cyprinid herpesvirus 2 (CyHV-2) is the etiological agent of Goldfish herpesviral haematopoietic necrosis (GHVHN) in goldfish. In this study, three new cell lines including Fantail goldfish gill (FtGG), Fantail goldfish liver (FtGL) and Fantail goldfish brain (FtGB) had been established and characterized from the gill, liver and brain tissue of C. auratus respectively. Cell lines were optimally grown at 28 °C in Leibovitz-15 (L-15) medium supplemented with 10 % fetal bovine serum (FBS). The PDT during exponential growth of FtGG, FtGL and FtGB cells were determined to be 41.47 h, 63.43 h and 79.79 h respectively. Karyotyping analysis of cell lines remained diploid (2n = 100). The revival rate was 82 %, 72 % and 70 % in FtGG, FtGL and FtGB cells respectively after 6 months of cryopreservation. All the three cells showed similar cytopathic effect (CPE) between 3–5 days post-infection (dpi) with CyHV-2 and complete destruction of the monolayer was observed at 8–10 dpi. The viral titers of CyHV-2 in FtGG, FtGL and FtGB reached 107.375±0.35 TCID50 ml−1, 104·55±0.070 TCID50 ml−1 and 106.45±0.070 TCID50 ml−1 respectively. These newly established cell lines will be a useful diagnostic tool for viral diseases in this fish species and also for the isolation and study of goldfish viruses in future.