龍膽石斑魚在經濟和營養價值上有著很高的價值，但也因其極容易受神經壞死病毒 （Nervous Necrosis Virus, NNV）感染，引起高達90%-100%的死亡率。雖然目前已經具有相對的疫苗，但，疫苗與溫度之間的關係還並不瞭解。因此爲了探討疫苗及不同溫度在對抗病毒感染的效果，本研究使用了年幼龍膽石斑魚 （ Epinephelus lanceolatus ）為實驗對象，再以本實驗室開發出對抗神經壞死病毒的重組單位疫苗, NNVCP-S5于三種不同的溫度，20°C、28°C和36°C，以腹腔注射的方式在龍膽石斑魚上使用疫苗，來測試免疫效益,另外以不同濃度的口服疫苗喂食龍膽石斑魚。再也以qPCR 來分析重組疫苗所會誘發的免疫相關的基因表現。實驗結果發現注射與口服疫苗會誘導Th 2免疫路徑基因，GATA-3與c-Maf表現的上升；Th 1免疫路徑基因，T-bet的表現下降，進而偏向Th2免疫路徑以產生免疫反應。在疫苗效力測試部分，RPS （relative percentage of survival）于NNV病毒106 TCID/mL攻毒下，高溫36°C下為57.13%，而在低溫，20°C為11.43%。實驗結果顯示于高溫下，疫苗效力為較好, 亦可以減低死亡率。因此，本實驗發現在高溫下，NNVCP-S5E疫苗能夠提供為龍膽石斑魚更有效的保護，也提供魚業界一個更好的使用疫苗，提供保護效果的方法。

rouper fish is having a very high value economically and also nutritionally, these make them a very important fish species in Asia. However, the grouper fish often suffers from a very mortality rate, up to 90%-100% when they are infected with Nervous Necrosis Virus (NNV). However, the effects of environmental temperatures on the vaccine efficacy is still not understood yet. With the aim to investigate the efficacy of vaccine against viral infection under different temperatures, juvenile giant grouper (Epinephelus lanceolatus) is used as a test subject in the study, NNVCP-S5E, a NNV vaccine that was developed by our lab previously was tested at three different temperatures, 20°C, 28°C and 36°C. Fishes were vaccinated through the method of injection, and oral vaccination was done with different dosages of vaccine, qPCR was done to investigate whether Th 1 or Th 2 pathway was induced by the NNVCP-S5E. The results from the study shows that both injection and oral vaccination with NNVCP-S5E will tend to direct the immune pathway towards Th2 pathway, as the expressions of gene markers GATA-3 and c-Maf was upregeulated; the expression of Th 1 pathway gene marker, T-bet was downregulated. For the vaccine efficacy tests under different temperatures, virus challenge with 106 TCID/mL gave a RPS, (relative percentage of survival) of 57.13% at 36°C; while 11.43% at 20°C. The results in this study shows that the vaccine efficacy is better at higher temperature, 36°C when compared to the lower temperature, 20°C, giving a better protection to the juvenile giant grouper fishes against viral infection. Thus, we can provide an ideal condition to the fisheries for a better vaccine efficacy and protection.

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