Multiple Passages of Grunt Fin Cells Persistently Infected with Red Seabream Iridovirus (RSIV) at 15ºC or 30ºC to Yield Uninfected Cells

Abstract

Red seabream iridovirus (RSIV), a member within genus Megalocytivirus (Iridoviridae), causes serious economic losses to marine fish aquaculture industry in East Asia. In this study, we established a Blue Striped Grunt Haemulon sciurus fin (grunt fin; GF) cell line persistently infected with RSIV (PI-GF^RSIV) by subculturing GF cells that survived RSIV inoculation. PI-GF^RSIV cells were morphologically indistinguishable from naive GF cells. They could stably produce RSIV at approximately 10^{4.9 ± 0.5} genomes per microliter after 24 passages over 18 months. The optimum temperature to produce RSIV in PI-GF^RSIV cells was 25°C. These cells also produced RSIV at 15, 20, and 30°C with multiple subcultures. The amount of RSIV yielded from PI-GF^RSIV cells decreased gradually by multiple subculturing at 15°C or 30°C. Red seabream iridovirus was no longer detected from PI-GF^RSIV cells after subcultures at these temperatures. These PI-GF^RSIV cells freed from RSIV infection exhibited a level of RSIV productivity similar to those of naive GF cells after inoculation with RSIV. Therefore, we consider that these PI-GF^RSIV cells were no longer infected with RSIV after multiple subculturing at 15°C or 30°C.

Received October 15, 2015; accepted June 27, 2016Published online October 13, 2016

Acknowledgments

This research was supported by a grant (NRF-2015R1D1A3A01016558) of the Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education, South Korea.