Influence of temperature on Mx gene expression profiles and the protection of sevenband grouper, Epinephelus septemfasciatus, against red-spotted grouper nervous necrosis virus (RGNNV) infection after poly (I:C) injection

Influence of temperature on the susceptibility of fish against virus infection has been studied for a decade. Recent reports have been shown the effects of rearing temperatures on the fish immune system against virus infection. However, the roles of temperature in regulation of type I interferon (IFN) system has not yet been investigated. Thus, the effects of temperature on type I IFN response were investigated in this study using poly (I:C) injection in sevenband grouper and Mx gene was used as a marker for type I IFN expression. Quantitative real-time PCR (qPCR) result showed that Mx expression profiles were moderately different between temperatures. The highly up-regulated Mx transcripts at 3 h post injection (hpi) were observed in high temperatures (25 °C and 30 °C) but not in low temperatures (15 °C and 20 °C). Meanwhile, low temperatures (15 °C and 20 °C) could detect the highly up-regulated Mx transcripts at 24 hpi. Expression of Mx transcripts was also observed at 72 hpi at 15 °C. Poly (I:C)-injected fish were challenged with RGNNV after 72 and 168 hpi. At 72 hpi, 100% of fish survived at all temperatures, whereas 95% survival rate was observed at 168 hpi at 25 °C during 14 days of observation. To further verify the duration period of an antiviral state at different temperatures, qPCR and endpoint dilution assay were used to quantify the number of virus in fish challenged with RGNNV. The reduction of viral copy numbers and viral titers could be observed at 72 and 168 hpi. However, high viral copy numbers and viral titers could be detected at 168 hpi at 30 °C. These results demonstrate that temperatures influenced on the Mx expression profiles and the duration period of an antiviral state efficiently interfered with virus replication at different temperatures.