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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 齊肖琪 | |
dc.contributor.author | Kun-Chan Tsai | en |
dc.contributor.author | 蔡昆展 | zh_TW |
dc.date.accessioned | 2021-06-15T01:40:27Z | - |
dc.date.available | 2011-07-16 | |
dc.date.copyright | 2009-07-16 | |
dc.date.issued | 2009 | |
dc.date.submitted | 2009-07-14 | |
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dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/43167 | - |
dc.description.abstract | 魚類結病毒(Betanodavirus)是造成全球海水養殖魚苗罹患神經壞死症之病原體,根據鞘蛋白質基因序列可分成Red-spotted Grouper Nervous Necrosis Virus(RGNNV)、Striped Jack(SJ)NNV、Barfin Flounder(BF)NNV,以及Tiger Puffer(TP)NNV四種基因型,其中RGNNV基因型寄主範圍及分佈地區最廣。本研究擬針對台灣周邊海域野生魚,以RT-PCR進行帶原NNV的篩檢,並利用分子演化學相關軟體分析該病毒分子流行病學的特徵。比對野生魚NNV分離株的鞘蛋白基因序列,發現台灣周邊海域野生魚帶原SJNNV及RGNNV,分佈地區以台灣北部海域為主,其中有4目12科的魚類首次發現為NNV寄主。根據基因交流的分析結果顯示,台灣周邊海域的RGNNV族群會經由水平傳播的方式影響養殖魚,再加上兩病毒族群均受到強烈正向天擇力量的影響,顯示未來可能有新變種病毒出現。根據病毒序列變異分佈的計算結果,台灣與中國目前是太平洋西岸最主要的NNV傳播中心。過去十年,台灣養殖魚的NNV分離株皆只有RGNNV基因型,不曾在爆發疫情的養殖魚中發現SJNNV,但在本研究中,台灣沿海野生魚帶原SJNNV的機率卻大於RGNNV數量。將台灣沿海帶原魚的SJNNV鞘蛋白質基因序列,和日本及歐洲養殖發病魚分離出的SJNNV序列進行比對,結果台灣的SJNNV自成一枝序群,日本和歐洲合成另一枝序群,顯示造成這兩群的不是地理隔絕的因素,反而可能是帶原魚與發病魚的差異;此外,台灣野生帶原魚的SJNNV枝序群不受天擇力量的影響,但日本和歐洲養殖發病魚的SJNNV枝序群卻受負向天擇影響,顯示決定SJNNV病原性有無的遺傳機制是非常保守的。 | zh_TW |
dc.description.abstract | Betanodaviruses, the causative agents of viral nervous necrosis disease, are classified into 4 genotypes, including red-spotted (RG), striped jack (SJ), barfin flounder (BF) and tiger puffer (TP) nervous necrosis virus (NNV). The aim of this study is to screen NNV in the wild fish collected from different aquatic area around Taiwan, and investigated the molecular phylogeny and epidemiology of those NNV isolates. Nested RT- PCR was applied for the screening of T4 segment of coat protein gene of NNV. Both RGNNV and SJNNV were detected in the wild fish in Taiwan, and the fish of 12 species of 4 orders were first reported to be NNV host. Strong positive selection was observed in RGNNV populations from both wild-field and cultured fish, indicating that the environment preferred to keep the mutations of RGNNV. Unequal gene flow, from trash fish to the seashore fish and then to cultured fish, was monitored in RGNNV populations. It is therefore suggested that the carrier fish or sea water played a role of RGNNV reservoir and might transmit the virus from sea area to land pond. The phylogenetic tree of SJNNV revealed that all isolates from carrier fish of Taiwan were clustered into one lineage, and all isolates from diseased fish in Japan and Europe were clustered into another lineage, indicating that the distinction was not due to geographic barrier. Negative evolution was revealed in SJNNV population of cultured fish in Japan and Europe, and neutral evolution was found in SJNNV from wild-field carrier fish around Taiwan, indicating that mutation of SJNNV would reduce viral pathogenecity on cultured fish, but would not affect the fitness in wild carrier fish. It is therefore suggested the SJNNV population would be less harm than RGNNV on aquatic industry. | en |
dc.description.provenance | Made available in DSpace on 2021-06-15T01:40:27Z (GMT). No. of bitstreams: 1 ntu-98-R96b41024-1.pdf: 738710 bytes, checksum: 80630fb0f0dcb7ae505774d0d4ddb517 (MD5) Previous issue date: 2009 | en |
dc.description.tableofcontents | 目錄
目錄………………………………………………………………………………………………………………………Ⅰ 圖目錄……………………………………………………………………………………………………………………Ⅴ 表目錄……………………………………………………………………………………………………………………Ⅵ 摘要………………………………………………………………………………………………………………………..Ⅷ Abstract…………………………………………………………………………………………………………………..Ⅸ 第一章 文獻回顧 1. 神經壞死症病毒介紹……………………………………………………………………………………..1 2. 神經壞死症病毒基因型分類………………………………………………………………………….1 3. 神經壞死症病毒寄主感染模式與傳播途徑…………………………………………………..2 4. 神經壞死症病毒分子流行病學及演化模式…………………………………………………..4 第二章 材料與方法 1. 野生魚採樣…………………………………………………………………………………………………….6 2. 核酸粹取……………………………………………………………………………………………………..…6 3. 反轉錄 - 聚合酶連鎖反應(RT - PCE) 3.1 反轉錄(RT)……………………………………………………………………………………….7 3.2 聚合酶連鎖反應(Polymerase chain reaction, PCR)………………………….7 3.3 巢氏聚合酶連鎖反應(nested - PCR)………………………………………………..7 4. 基因選殖(gene cloning)…………………………………………………………………………………8 5. 病毒核酸序列分析(Sequence analysis)……………………………………………………….8 6. 分子演化生物學軟體計算 6.1 序列特性分析………………………….………………………..………………………………9 6.2 遺傳多樣性及遺傳結構………..…………………………………………………………10 6.3 族群變動分析…………..……………………………………………………………………..10 6.4 天擇力量測驗…………..……………………………………………………………………..11 6.5 族群間基因流方向…..……………………………………………………………………..12 6.6 演化速率估算…………..……………………………………………………………………..12 第三章 結果 1. 台灣野生魚帶原SJNNV及RGNNV………………………………………………………….13 1.1 下雜魚類SJNNV檢出率高於RGNNV檢出率………………………………13 1.2 沿岸魚類SJNNV檢出率近於RGNNV檢出率………………………………13 2. 台灣不同水域中RGNNV及SJNNV分離株的親緣關係分析………………….…13 2.1 同一基因NNV分離株間之基因交流不受寄主分佈水域的地緣限制………………………………………………………………….…………………………….…..13 2.1.1 RGNNV………………………………..……………………………………………….13 2.1.2 SJNNV…………………………………………………………………………...…….14 2.2 台灣SJNNV分離株遺傳變異源自一核心……………………………………..15 2.3 NNV族群歷史動態分析(Historical Demographic analysis)……………16 2.3.1 養殖魚的RGNNV族群曾經歷族群擴張,下雜魚的RGNNV族群則相對穩定………………………………………………………………….16 2.3.2 下雜魚的SJNNV族群曾歷經族群擴張………………….……….….17 2.4族群天擇力量測驗(Analysis of Natural Selection Forces)………………18 2.4.1 RGNNV及SJNNV族群皆受到正向天擇力量影響……………..18 2.4.2 發病魚RGNNV族群T4序列不同胺基酸位置受不同天擇力量影響..................................................................................................18 2.4.3 台灣SJNNV族群受到單一的天擇力量影響………….……………….19 2.5 族群分化及基因交流評估(Population Divergence and Gene Flow)………………………………………………………………………………………………19 2.5.1 野生魚的RGNNV可能經由不對等基因交流影響養殖魚………………………………..……………………………………………………….19 2.5.2 日本野生魚SJNNV族群向南移動可能影響台灣野生魚族群帶原SJNNV………………..…….…………………….………………………….19 2.6 NNV T4區域核酸具有高演化速率……………………………………..…………20 3.全球地區RGNNV及SJNNV核酸序列的親緣關係……………………………………..20 3.1 遺傳變異分析(Genetic Diversity)及親緣關係重建(Phylogeny Reconstruction)……..……………………………………….………………………………………20 3.1.1全球RGNNV族群與各自地理區無相對應的關係……….…….…20 3.1.2全世界SJNNV族群的分群與致病性並無關連性……………..…..21 3.2 台灣、日本與歐洲的SJNNV族群皆歷經族群擴張………..………...……21 3.3 RGNNV經由台灣與大陸沿海養殖石斑苗的出口而外移………………22 3.4 自病魚分離的SJNNV受到負向天擇影響….……….……………….………..22 第四章 討論 1. 野生魚感染帶原狀況……………………………………………………………………………………24 2. 台灣RGNNV族群的分子流行病學模式……………………………………………………….24 3. 台灣SJNNV族群的分子流行病學模式…………………………………………………………26 4. 全球RGNNV族群的分子流行病學模式……………………………………………………….27 5. 全球SJNNV族群的分子流行病學模式…………………………………………………………28 第五章 參考文獻…………………………………………………………………………………………………30 附錄一、下雜魚採集名單及採集數量……………………………………………….…………………..71 附錄二、沿岸魚採集名單及採集數量………………………………………………………….………..76 附錄三、本研究中所使用資料庫RGNNV的序列資料………………………………………….80 附錄四、本研究中所使用資料庫SJNNV的序列資料…………………………………………..84 附錄五、全球NNV感染魚類宿主之名錄…………………………………………………………………85 | |
dc.language.iso | zh-TW | |
dc.title | 魚類神經壞死病毒之分子演化與分子流行病學探討 | zh_TW |
dc.title | Molecular phylogeny and epidemiology of Nervous Necrosis Virus | en |
dc.type | Thesis | |
dc.date.schoolyear | 97-2 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 張繼堯,林弘都,王弘毅,林思民 | |
dc.subject.keyword | NNV,RGNNV,SJNNV,分子演化,野生帶原魚,正向天擇,自然儲所, | zh_TW |
dc.subject.keyword | NNV,RGNNV,SJNNV,molecular evolution,wild carrier fish,positive selection,natural reservoir, | en |
dc.relation.page | 94 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2009-07-14 | |
dc.contributor.author-college | 生命科學院 | zh_TW |
dc.contributor.author-dept | 動物學研究所 | zh_TW |
顯示於系所單位: | 動物學研究所 |
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