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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 童心欣(Hsin-Hsin Tung) | |
dc.contributor.author | Tzu-Hao Chen | en |
dc.contributor.author | 陳子豪 | zh_TW |
dc.date.accessioned | 2021-06-16T17:26:07Z | - |
dc.date.available | 2015-09-24 | |
dc.date.copyright | 2012-08-28 | |
dc.date.issued | 2012 | |
dc.date.submitted | 2012-08-16 | |
dc.identifier.citation | Adela Yanez, M., Nocker, A., Soria-Soria, E., Murtula, R., Martinez, L., and Catalan, V. (2011). Quantification of viable Legionella pneumophila cells using propidium monoazide combined with quantitative PCR. J Microbiol Methods 85, 124-130.
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dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/64003 | - |
dc.description.abstract | 北勢溪為台北集水區重要水源之一,此區域自水源發源地經坪林地區與坪林處理廠,最後流入翡翠水庫。我們針對人口密集區與污水廠前後採集水樣,藉由了解微生物群落的消長,顯示出此區域的汙染情況。
焦磷酸定序(pyrosequencing)為次世代高通量定序方法之一,近年來常用於研究環境之細菌群落變化。為了偵測水體真實存活的細胞,使用碘化丙啶(propidium monoazide, PMA),其為一種DNA螯合染劑,在照光下能與死亡細胞之DNA結合,並抑制聚合酶鏈鎖反應(polymerase chain reaction, PCR)結果。 本研究以焦磷酸定序法分析台北水源集水區之北勢溪五個測站的細菌群落,並分為兩個部分。(1)針對所有細菌(16S rRNA gene之V1-V2超高變異區間)水體樣品並結合PMA方法偵測活/死細胞進行探討。(2)針對與擬桿菌門(Bacteroidetes)比較水體樣品與糞便樣品之異同。結果發現,水樣細菌群落中,Proteobacteria、Fusobacteria與Bacteroidetes各占整體水樣16.43-43.36%、0.84-19.77% 與 2.30-60.21%。PMA前處理與無PMA前處理細菌群落有小幅度差異,PMA前處理之樣品之物種豐富度大於無PMA前處理之樣品,推測原因為PMA的前處理會抑制一些優勢物種的增幅,導致被遮蔽的物種顯露出來。PMA前處理所偵測到之Enterobacteriaceae、Bacteroides、Arcobacter、Rickettsia等菌群可能與人口密集相關。 針對Bacteroidetes的部分,利用水樣與糞便樣品進行OTU交互分析,OTU交互分析是以水樣與糞便樣品以文氏圖做交集計算,藉由交集多寡表示河川污染情況。結果顯示,集水區河川經人口密集區後,水體中所測得之動物糞便交集比例從1.5%上升至18.59%;經污水廠後的坪林污水下游測站則下降至8.89%,其中以人類糞便之比例變化最顯著。經過資料庫比對後,發現這可能為Prevotella copri、Bacteroides plebeius等人類腸道存在之菌種。根據結果,藉由OTU交互分析可以了解河川之汙染程度。 | zh_TW |
dc.description.abstract | Bei-Shih river is one of the raw water sources for Taipei metropolitan area. It flows through Ping-lin area, Ping-lin wastewater treatment plant, and enter the Fei-Tsui reservoir. Samples were collected at the upstream and downstream of the highly populated area and downstream of the Ping-lin wastewater treatment plant to investigate the bacterial community variation by pyrosequencing.
The purpose of this study was to target the hypervariable regions V1-V2 of the 16S rRNA gene and investigate the bacterial communities from Bei-Shih river by combination of PMA treatment and pyrosequencing. Also we compared the Bacteroidales group with fecal sources and freshwater. In our results, the phyla Proteobacteria, Fusobacteria and Bacteroidetes in freshwater were 16.43-43.36%, 0.84-19.77% and 2.30-60.21%, respectively. The bacterial community profiles of PMA-treated sample and non-PMA-treated sample were slightly different. The PMA-treated samples had more OTUs than non-PMA-treated samples. The gourps of Enterobacteriaceae, Bacteroides, Arcobacter, Rickettsia, were found in PMA-treamted water samples, and the ratio of each group to total bacteria was positively correlated to total coliforms. The correlation of shared OTUs between Bacteroidales communities of water samples and fecal samples were calculated. The results showed that the percentage correlation in Wan-Tan was 1.5% and increased to 18.59% in highly populated area. The shared OTUs of Prevotella copri and Bacteroides plebeius species, which related to human intestine bacteria, can also be found in freshwater. As a result, the degree of contamination in a region can be analyzed by the percentage of total and shared OTUs. | en |
dc.description.provenance | Made available in DSpace on 2021-06-16T17:26:07Z (GMT). No. of bitstreams: 1 ntu-101-R99541106-1.pdf: 3025893 bytes, checksum: 60dd3e04f01cb91b6711e20014b5df3d (MD5) Previous issue date: 2012 | en |
dc.description.tableofcontents | 誌謝 i
摘要 iii Abstract v 總目錄 vii 圖目錄 x 表目錄 xi 第一章 前言 1 第二章 文獻回顧 3 2.1 環境與細菌群落 3 2.2 相關定序技術之發展 4 2.2.1 Sanger定序技術 4 2.2.2 高通量定序技術──焦磷酸定序 5 2.3 完整細胞偵測的方法介紹 7 2.4 指標微生物(Indicator bacteria)與微生物追蹤法 9 2.4.1 指標微生物介紹 9 2.4.2 指標微生物的限制 9 2.4.3 微生物追蹤法的介紹 10 2.4.4 宿主專一性聚合酶連鎖反應原理 10 2.4.5 宿主專一性聚合酶連鎖反應的限制 11 第三章 實驗方法與步驟 13 3.1 實驗架構 13 3.2 採集地點介紹 15 3.3 水樣採集與濃縮 18 3.4 PMA處理方法 19 3.5 DNA萃取方法 20 3.5.1 DNA萃取藥品配製與準備 20 3.6 糞便樣品的處理 22 3.7 聚合酶連鎖反應(PCR) 22 3.8 瓊脂膠體電泳分析 24 3.9 瓊脂膠體萃取 25 3.10 DT-PCR 25 3.11 DNA定量 29 3.12 焦磷酸定序 29 3.13 高通量定序資料處理 31 3.13.1 定序資料之前處理—修剪、分類、對齊 31 3.12.2 OTU計算與稀釋曲線 33 3.13.3 α-多樣性指數之計算 33 3.13.4 β-多樣性之計算-UniFrac與UPGMA、PCoA 35 3.13.5 群落分析 38 3.13.6 水樣與糞便樣品之OTU交互分析 38 第四章 結果與討論 39 4.1 α-多樣性相關分析與討論 39 4.2 β-多樣性之結果 44 4.3 細菌群落之菌群討論 49 4.4 水樣與糞便樣品之OTU交互分析結果 59 4.4.1 河川上下游與未PMA前處理處理之討論 63 4.4.2 河川上下游與PMA處理之討論 64 4.4.3可能因人口密集區而細菌比例上升之族群 65 第五章 結論 68 第六章 未來方向與建議 70 參考文獻 71 附錄 78 | |
dc.language.iso | zh-TW | |
dc.title | 以焦磷酸定序技術探討北勢溪細菌菌相組成 | zh_TW |
dc.title | Analysis of Bacteria in Bei-Shih River by Pyrosequencing | en |
dc.type | Thesis | |
dc.date.schoolyear | 100-2 | |
dc.description.degree | 碩士 | |
dc.contributor.coadvisor | 湯森林(San-Lin Tang) | |
dc.contributor.oralexamcommittee | 陳俊堯,郭志鴻 | |
dc.subject.keyword | 高通量定序,焦磷酸定序,擬桿菌門,細菌群落,碘化丙啶,台北水源集水區, | zh_TW |
dc.subject.keyword | high throughput sequencing,pyrosequencing,Bacteroidetes,bacterial community,propidium monoazide(PMA),Taipei water source catchment, | en |
dc.relation.page | 82 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2012-08-16 | |
dc.contributor.author-college | 工學院 | zh_TW |
dc.contributor.author-dept | 環境工程學研究所 | zh_TW |
顯示於系所單位: | 環境工程學研究所 |
文件中的檔案:
檔案 | 大小 | 格式 | |
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ntu-101-1.pdf 目前未授權公開取用 | 2.95 MB | Adobe PDF |
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