title: 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-GFRSIV) by subculturing GF cells that survived RSIV inoculation. PI-GFRSIVcells were morphologically indistinguishable from naive GF cells. They could stably produce RSIV at approximately 104.9 ± 0.5genomes per microliter after 24 passages over 18 months. The optimum temperature to produce RSIV in PI-GFRSIVcells was 25°C. These cells also produced RSIV at 15, 20, and 30°C with multiple subcultures. The amount of RSIV yielded from PI-GFRSIVcells decreased gradually by multiple subculturing at 15°C or 30°C. Red seabream iridovirus was no longer detected from PI-GFRSIVcells after subcultures at these temperatures. These PI-GFRSIVcells 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-GFRSIVcells were no longer infected with RSIV after multiple subculturing at 15°C or 30°C. Received October 15, 2015; accepted June 27, 2016.
authors.affiliation: a Department of Aqualife Medicine , Chonnam National University , Daehak-ro 50, Yeosu 59626 , South Korea.
authors.forename: So-Young
authors.initials: SY
authors.lastname: Oh
authors.affiliation: a Department of Aqualife Medicine , Chonnam National University , Daehak-ro 50, Yeosu 59626 , South Korea.
authors.forename: Toyohiko
authors.initials: T
authors.lastname: Nishizawa
mesh_terms: D058500:Animal Fins / Q000821:virology*; D000818:Animals; D018929:Cell Culture Techniques; D002460:Cell Line; D004266:DNA Virus Infections / Q000662:veterinary* / Q000821:virology; D005393:Fish Diseases / Q000821:virology*; D007497:Iridoviridae / Q000302:isolation & purification*; D010473:Perciformes; D013696:Temperature