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Please use this identifier to cite or link to this item: http://ntour.ntou.edu.tw:8080/ir/handle/987654321/34564

Title: 點帶石斑口服次單位疫苗之效果評估
The Evaluation of the Oral Subunit Vaccine (Grouper Nervous Necrosis Virus Coat Protein) Against Orange Spotted Groupers (Epinephelus coioides) Infected by NNV
Authors: Chen-Yen Choong
Contributors: NTOU:Department of Aquaculture
Keywords: 病毒神經壞死症;重組外鞘蛋白;次單位疫苗
Viral nervous necrosis;recombinant NNV coat proteins;Subunit Vaccine
Date: 2013
Issue Date: 2013-10-07T02:42:37Z
Abstract: 病毒神經壞死症(Viral nervous necrosis)每年在台灣均造成養殖業重大的經濟損失,主要感染石斑魚苗及幼苗,並引起高達80 % - 100 %的死亡率。為了因應神經壞死症對養殖業造成的損失,本研究主要發展NNV次單位疫苗,並以口服的方式給予魚苗免疫,期望達到對抗NNV感染之目的。本實驗利用重組蛋白的方式把NNV之外鞘蛋白基因轉接到表現載體pET 23a中,並由E. coli BL21(DE3) plysS表現系統製造出NNV重組外鞘蛋白,最後經褐藻酸鈉-幾丁質或洋菜膠包覆後,進行石斑魚投餵實驗。投餵一星期疫苗,免疫14天,再以6 × 108 TCID50 / mL病毒劑量進行攻擊實驗;以相對存活率(Relative Percentage Survival, RPS)來評估疫苗的保護效果。褐藻酸鈉-幾丁質包覆組中,以每公斤飼料添加0.1 g次單位疫苗,RPS達50%;而每公斤飼料添加1 g次單位疫苗,RPS達60%。在洋菜膠包覆組中,疫苗有無包覆並無明顯差異,RPS均為50 %。
Nerve necrosis is a major viral disease which infects groupers and other fish in their larvae stage. The mortality rate of infected groupers and larvae reaches approximately 80-100%, resulting in a great economic loss for the aquaculture of Taiwan. In order to prevent a possible outbreak, the development of vaccine is essential to the survival of groupers in aquaculture throughout Taiwan. In this study, an experimental Nervous Necrosis Virus (NNV) vaccine was developed to prevent infection by introducing it orally to grouper larvae. The oral subunit NNV vaccines, NNV coat proteins expressed by E. coli BL21(DE3) plysS, were encapsulated with sodium alginate-chitin and with agar respectively. After applying the vaccine continuously for a week, we halt the immunization of the grouper for 14 days before injecting 6 X 108 TCID50 / mL of NNV into the abdomen of the fish. In order to determine the results, we used calculated the relative percentage survival (RPS) of the subjects after being treated with the vaccine. The Sodium alginate-chitin encapsulated groups showed two different RPS. The one that contained 1g recombinant NNV coat proteins per 1kg formula showed a 60% RPS. While 0.1g recombinant NNV coat proteins per 1kg formula showed a 50% RPS. In the agar encapsulated groups, 1g and 0.1g recombinant NNV coat proteins per 1 kg formula both showed a 50% RPS.
URI: http://ethesys.lib.ntou.edu.tw/cdrfb3/record/#G0019933051
Appears in Collections:[水產養殖學系] 博碩士論文

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