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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 吳育騏 | zh_TW |
| dc.contributor.advisor | Yu-Chi Wu | en |
| dc.contributor.author | 尹建翔 | zh_TW |
| dc.contributor.author | Chien-Hsiang Yin | en |
| dc.date.accessioned | 2024-12-24T16:31:45Z | - |
| dc.date.available | 2024-12-25 | - |
| dc.date.copyright | 2024-12-24 | - |
| dc.date.issued | 2024 | - |
| dc.date.submitted | 2024-11-28 | - |
| dc.identifier.citation | Abós, B., Bailey, C., Tafalla, C., 2022. Adaptive immunity, Principles of Fish Immunology: From Cells and Molecules to Host Protection. Springer, pp. 105-140.
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/96360 | - |
| dc.description.abstract | 石斑魚為亞洲地區水產養殖產業之重要經濟魚種。神經壞死症病毒 (Nervous necrosis virus, NNV)及石斑魚虹彩病毒(Grouper iridovirus, GIV)為石斑魚主要的兩種病毒性病原，感染後會導致魚隻大量死亡。NNV在分類上屬於野田病毒科(Nodaviridae)的 β-野田病毒屬(而GIV在分類上屬於虹彩病毒科(Iridoviridae)的蛙病毒屬(Ranavirus)。接種疫苗是預防病毒性疾病的有效策略之一，且雙價疫苗能降低接種疫苗時所耗費的人力與時間成本及免疫期間魚隻受緊迫的次數。本篇研究之目的為開發NNV-GIV雙價疫苗來保護石斑魚抵抗NNV與GIV，並測試NNV-GIV雙價疫苗能否交叉保護魚隻抵抗嘉魶魚虹彩病毒(Red sea bream iridovirus, RSIV)。安全性測試結果顯示，接種NNV-GIV雙價疫苗的魚隻皆活著且食慾正常且免疫魚隻的臟器均未出現組織病變之情況，說明此疫苗對魚隻具有安全性。攻毒試驗結果顯示以NNV及GIV進行攻毒後的免疫魚隻之相對存活率分別為85.1%及86.8%。ELISA結果顯示接種NNV-GIV雙價疫苗能顯著提升魚隻血清中抗NNV、GIV與RSIV之專一性抗體力價。中和試驗結果顯示，接種NNV-GIV雙價疫苗能顯著提升魚隻血清中抗NNV與GIV之中和抗體力價。免疫魚隻的脾臟與頭腎中免疫相關基因的表現量皆有顯著提升，這些免疫相關基因包括TNF-α、IL1-β、MHC-I、MHC-II、CD4、CD8、IgM、IgT、Mx，此結果顯示NNV-GIV雙價疫苗能成功誘發魚隻體內的細胞免疫與體液免疫反應。總結上述研究結果，NNV-GIV雙價疫苗可以有效提升石斑魚的免疫力，保護魚隻對抗病毒的感染。 | zh_TW |
| dc.description.abstract | Grouper is an economically important fish species in aquaculture industry in Asia.Nervous necrosis virus (NNV) and grouper iridovirus (GIV) are two major viral pathogens in groupers and have caused massive mortality. NNV belongs to Betanodavirus genus of Nodaviridae family, and GIV classified into Ranavirus genus of Iridoviridae family. Vaccination is an effective strategy to prevent viral disease, and bivalent vaccines can reduce the cost of both labor and time and decrease the times of stress during vaccination. The purpose of this study was to develop a bivalent vaccine to protect groupers against NNV and GIV, and to investigate whether the NNV-GIV bivalent vaccine could have cross-protection against red sea bream iridovirus (RSIV). The safety test revealed that fish immunized with NNV-GIV bivalent vaccine were all alive and exhibited normal appetite. Moreover, the organs of immunized fish showed no histopathologic effect, indicating that the vaccine is safe for fish. In the challenge test, the relative percent survival (RPS) values of immunized fish were 85.1% and 86.8% in NNV and GIV challenge, respectively. The results of ELISA revealed that immunization with the NNV-GIV bivalent vaccine can significantly increase the titers of specific antibody against NNV, GIV and RSIV in fish sera. The neutralization test revealed that immunization with the NNV-GIV bivalent vaccine can significantly increase the titers of neutralizing antibody in fish sera against NNV and GIV. The expression of immune-related genes in the spleen and head kidney of immunized fish all significantly increased, and these genes includes TNF-α, IL1-β, MHC-I, MHC-II, CD4, CD8, IgM, IgT, and Mx. This result indicated that the NNV-GIV bivalent vaccine could effectively induce cellular and humoral immune responses in fish. In conclusion, the NNV-GIV bivalent vaccine can effectively improve the immunity of grouper and protect the fish against viral infection. | en |
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| dc.description.provenance | Made available in DSpace on 2024-12-24T16:31:45Z (GMT). No. of bitstreams: 0 | en |
| dc.description.tableofcontents | 目次
摘要 i Abstract ii 目次 iv 圖表次 vii 1 前言 1 1.1 石斑魚養殖 1 1.2 養殖石斑魚面臨的重大病毒性疾病 1 1.2.1 病毒性神經壞死症(viral nervous necrosis, VNN) 1 1.2.2 魚類虹彩病毒症 3 1.3 魚類的免疫系統 4 1.3.1 魚類的免疫器官 4 1.3.2 魚類的先天性免疫 5 1.3.2.1 細胞免疫 5 1.3.2.2 體液免疫 5 1.3.3 魚類的適應性免疫 6 1.3.3.1 細胞免疫 6 1.3.3.2 體液免疫 6 1.4 魚類疫苗 7 1.4.1 疫苗的製備方式 7 1.4.1.1 不活化疫苗 7 1.4.1.2 減毒疫苗 8 1.4.1.3 DNA疫苗 8 1.4.1.4 重組蛋白疫苗 8 1.4.1.5 類病毒顆粒疫苗 9 1.4.2 疫苗的免疫方式 9 1.4.2.1 注射免疫 9 1.4.2.2 浸泡免疫 9 1.4.2.3 口服免疫 10 1.5 研究目的 10 2 材料與方法 12 2.1 實驗魚 12 2.2 細胞株 12 2.3 病毒株 12 2.4 病毒生產條件測試 13 2.5 不活化病毒之製備 13 2.6 魚隻免疫 15 2.7 魚隻採樣 15 2.8 攻毒試驗 16 2.9 間接酵素連結免疫吸附試驗(indirect ELISA) 16 2.10 中和試驗 17 2.11 RNA萃取與反轉錄(reverse transcription) 18 2.12 即時聚合酶連鎖反應(real-time PCR) 18 2.13 疫苗中的病毒核酸檢測 19 3 結果 20 3.1 病毒生產條件測試 20 3.2 不活化病毒疫苗安全性測試 20 3.3 疫苗在魚體內的殘留情況 20 3.4 疫苗保護效果 21 3.5 免疫魚血清中之專一性抗體力價 22 3.6 免疫魚黏膜中之專一性抗體力價 23 3.7 免疫魚血清中之中和抗體力價 23 3.8 疫苗對免疫魚隻的免疫相關基因調控 24 3.9 不活化病毒之核酸檢測 25 4 討論 26 4.1 疫苗安全性 26 4.2 攻毒試驗 26 4.3 免疫魚血清中的專一性抗體 28 4.4免疫魚血清中的中和抗體 29 4.5 疫苗誘發免疫基因表現 30 4.6 病毒核酸誘發干擾素反應 31 4.7 結論與未來展望 32 參考文獻 33 圖表次 圖一、 NNV-GIV雙價疫苗免疫後之組織切片分析 42 圖二、 NNV-GIV雙價疫苗免疫後魚隻腹腔的疫苗殘留情況 44 圖三、 以 NNV-GIV雙價疫苗 免疫 之龍膽石斑經 NNV攻毒後之累積死亡率 45 圖四、 以 NNV-GIV雙價疫苗 免疫 之龍膽石斑經 GIV攻毒後之累積死亡率 46 圖五、 以 NNV-GIV雙價疫苗 免疫 之龍膽石斑 經 RSIV攻毒後之累積死亡率 47 圖六、以 ELISA檢測免疫組龍膽石斑在 免疫後 第 0、 1、 2、 3、 6週的血清中對NNV、 GIV與 RSIV之專一性抗體力價 48 圖七、以 ELISA檢測 對照 組龍膽石斑 經 GIV攻毒後存活 的 魚隻 血清中對 GIV與 RSIV之專一性抗體力價 49 圖八、以 ELISA檢測 免疫組與 對照組龍 虎 斑在 免疫後 第 6週的 黏膜與 血清中對NNV之專一性抗體力價 50 圖九、以 ELISA檢測 免疫組與 對照組龍 虎 斑在 免疫後 第 6週的 黏膜與 血清中對GIV之專一性抗體力價 51 圖十、以 ELISA檢測 免疫組與 對照組龍 虎 斑在 免疫後 第 6週的 黏膜與 血清中對RSIV之專一性抗體力價 52 圖十一、以中和試驗檢測免疫組龍膽石斑在 免疫後 第 0週與第 6週的血清中對NNV、 GIV與 RSIV之中和抗體力價 53 圖十二 、 NNV-GIV雙價疫苗對龍膽石斑 TNF-α基因之調控 54 圖十三 、 NNV-GIV雙價疫苗對龍膽石斑 IL-1β基因之調控 55 圖十四、 NNV-GIV雙價疫苗對龍膽石斑 MHC-I基因之調控 56 圖十五、 NNV-GIV雙價疫苗對龍膽石斑 MHC-II基因之調控 57 圖十六、 NNV-GIV雙價疫苗對龍膽石斑 CD4基因之調控 58 圖十七、 NNV-GIV雙價疫苗對龍膽石斑 CD8基因之調控 59 圖十八、 NNV-GIV雙價疫苗對龍膽石斑 IgM基因之調控 60 圖十九、NNV-GIV雙價疫苗對龍膽石斑雙價疫苗對龍膽石斑IgT基因之調控基因之調控 61 圖二十、NNV-GIV雙價疫苗對龍膽石斑雙價疫苗對龍膽石斑Mx基因之調控基因之調控 62 圖二一、不活化不活化NNV及不活化及不活化GIV之核酸檢測之核酸檢測 63 表一、GIV複製最佳條件測試複製最佳條件測試 64 表二、qPCR所使用的引子所使用的引子(primer)序列序列 65 | - |
| dc.language.iso | zh_TW | - |
| dc.subject | 石斑魚虹彩病毒 | zh_TW |
| dc.subject | 抗體 | zh_TW |
| dc.subject | 體液免疫 | zh_TW |
| dc.subject | 細胞免疫 | zh_TW |
| dc.subject | 雙價不活化疫苗 | zh_TW |
| dc.subject | 神經壞死症病毒 | zh_TW |
| dc.subject | antibody | en |
| dc.subject | grouper iridovirus (GIV) | en |
| dc.subject | Nervous necrosis virus (NNV) | en |
| dc.subject | bivalent inactivated vaccine | en |
| dc.subject | cellular immunity | en |
| dc.subject | humoral immunity | en |
| dc.title | 神經壞死症病毒與石斑魚虹彩病毒雙價疫苗對石斑魚保護效力之研究 | zh_TW |
| dc.title | Study on protective efficacy of nervous necrosis virus and grouper iridovirus bivalent vaccine on groupers | en |
| dc.type | Thesis | - |
| dc.date.schoolyear | 113-1 | - |
| dc.description.degree | 碩士 | - |
| dc.contributor.oralexamcommittee | 韓玉山;陳立涵;胡鄴方;蓋玉軒 | zh_TW |
| dc.contributor.oralexamcommittee | Yu-San Han;Li-Han Chen;Yeh-Fang Hu;Yu-Hsuan Kai | en |
| dc.subject.keyword | 神經壞死症病毒,石斑魚虹彩病毒,雙價不活化疫苗,細胞免疫,體液免疫,抗體, | zh_TW |
| dc.subject.keyword | Nervous necrosis virus (NNV),grouper iridovirus (GIV),bivalent inactivated vaccine,cellular immunity,humoral immunity,antibody, | en |
| dc.relation.page | 65 | - |
| dc.identifier.doi | 10.6342/NTU202404631 | - |
| dc.rights.note | 同意授權(限校園內公開) | - |
| dc.date.accepted | 2024-11-28 | - |
| dc.contributor.author-college | 生命科學院 | - |
| dc.contributor.author-dept | 漁業科學研究所 | - |
| dc.date.embargo-lift | 2029-11-29 | - |
| 顯示於系所單位： | 漁業科學研究所 | |
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