| Summary: | 碩士 === 國立臺灣海洋大學 === 水產養殖學系 === 106 === Grouper is one of the most high economic value fish species in Taiwan, its history in Taiwanese aquaculture stands more than 30 years. However, the expansion of farms and rising stocking density has made various epidemic diseases become a big issue. This issue limits the development of the grouper fish breeding industry. Among many diseases, viral diseases are extremely threatening since that they are hard to be controlled with drugs. In all kinds of fish viral disease, the disease caused by Nervous Necrosis virus(NNV) and Grouper Iridovirus(GIV) are the most critical diseases in the grouper farm. In this study, siRNA and DNA vaccine were orally delivered into grouper fish and tested their protection against NNV and GIV virus. At the different time points, the serum, brain, intestine, spleen and head kidney of the grouper were collected to detect the production of antibodies and the gene expression by using ELISA and qPCR. Our results showed that the expression level of siRNA in intestine was highest at 20 days post of siRNA feeding. The gene expression of NNV showed that the siRNA could inhibit the replication of NNV. In addition, the results in ELISA analysis showed that DNA vaccine was able to induce antibody level against GIV in serum. By using qPCR, the results also showed that DNA vaccine could enhance the immune gene response in grouper and inhibit the replication of GIV. For example, by detecting the expression of MHC class II and CD4 gene in qPCR, the results in those with DNA vaccine had been increased. This proved that DNA vaccine is useful in defensing the attack from GIV. In general, in this study we find that by using both siRNA and DNA vaccine, it is able to improve the immune response against both NNV and GIV. We expect this study to be somewhat useful to bringing the oral delivery system into the grouper farm industry in the future.
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