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DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 童心欣(Hsin-hsin Tung) | |
dc.contributor.author | Yi-An Lin | en |
dc.contributor.author | 林益安 | zh_TW |
dc.date.accessioned | 2021-06-07T23:45:49Z | - |
dc.date.copyright | 2020-08-21 | |
dc.date.issued | 2020 | |
dc.date.submitted | 2020-08-10 | |
dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/16777 | - |
dc.description.abstract | 近年來工業發展迅速,重金屬污染案件層出不窮,已引起社會大眾密切關注。河口底泥為河川長期累積沖刷而成,其做為污染物之儲存庫,對重金屬具有沉澱、吸附與錯合作用使其移動性降低,進而成為潛在重金屬累積之熱點。重金屬污染除了造成環境菌群結構改變外,同時也極有可能是導致環境中的抗生素抗性基因(Antibiotic Resistance Genes, ARGs)傳播之關鍵因素。然而目前針對重金屬對菌群結構的改變及抗生素抗性基因於河口底泥的分佈尚未得到廣泛之研究,有鑑於此,本研究選擇新竹香山濕地及西南沿海一帶做為研究場址,透過採集表層河口底泥,以次世代定序技術分析菌群結構,以及使用即時聚合酶連鎖反應定量分析抗生素抗性基因,最後透過相關性分析找出重金屬與兩者之關聯。結果顯示河口底泥之抗生素抗性基因與Class 1 Integrons及重金屬具有顯著之關聯性,代表水平基因轉移及共選擇機制可能為河口底泥抗生素抗性基因傳播的主要原因。菌群結構部分,PCoA及CCA分析顯示即便重金屬濃度無超過底泥品質指標之分類管理及用途限制辦法之上限值,長期的污染仍然可能導致菌群結構產生變化,其中AB-539-J10、Desulfarculaceae、Hydrogenophilaceae、Marinilabiliaceae、Methanomicrobiaceae、Methanoregulaceae、Methanosaetaceae、SG8-4、Spirochaetaceae、Sulfurovaceae、Synergistaceae等11科菌科與多種重金屬顯著正相關,為河口底泥重金屬污染下之優勢菌科。最後透過網絡分析發現Chloroflexi、Euryarchaeota、Spirochaetes、Tenericutes為抗生素抗性基因與菌門網絡中之熱點,顯示其可能為抗生素抗性基因於河口底泥之潛在宿主。 | zh_TW |
dc.description.abstract | Industry has developed rapidly in recent years and heavy metal pollution cases have emerged one after another, which has aroused attention from the public. As a reservoir of pollutants, estuary sediments can reduce heavy metals’ mobility due to precipitation, adsorption and complexation. Therefore, they become hotspots of heavy metal pollution. In addition to changing the microbial community structure, heavy metals are also likely to be a critical factor that promote the propagation of antibiotic resistance genes (ARGs). However, impact of heavy metals on the changes in microbial community structure and the distribution of antibiotic resistance genes in estuary sediments have not been studied extensively. In view of this, this study collected surface estuary sediments from Hsinchu Xiangshan Wetland and the southwest coastal area of Taiwan. Microbial community structure was analyzed by next-generation sequencing technology, and abundance of antibiotic resistance genes was analyzed by real-time polymerase chain reaction. Results showed that antibiotic resistance genes had significant correlations with Class 1 Integrons and heavy metals, indicating that horizontal gene transfer and co-selection mechanisms may be the main reasons for the spread of antibiotic resistance genes in estuary sediments. For the microbial community structure, PCoA and CCA analysis showed that even if the heavy metal concentration did not exceed the upper limit of sediment quality regulations, long-term pollution could still change microbial community structure. Among all the families, AB-539-J10, Desulfarculaceae, Hydrogenophilaceae, Marinilabiliaceae, Methanomicrobiaceae, Methanoregulaceae, Methanosaetaceae, SG8-4. Spirochaetaceae, Sulfurovaceae and Synergistaceae had strong and positive correlations with multiple heavy metals, indicating that they are the dominant microorganisms in the estuary sediments under heavy metal pollution. At last, we performed network analysis and found that Chloroflexi, Euryarchaeota, Spirochaetes, and Tenericutes are hotspots in the network of antibiotic resistance genes and phylums, showing that they may be potential hosts for antibiotic resistance genes in estuary sediments. | en |
dc.description.provenance | Made available in DSpace on 2021-06-07T23:45:49Z (GMT). No. of bitstreams: 1 U0001-1008202011320800.pdf: 11311045 bytes, checksum: 4a7a51d515364e7faa718bf74d2d37a4 (MD5) Previous issue date: 2020 | en |
dc.description.tableofcontents | 目錄 誌謝 I 摘要 III ABSTRACT V 目錄 VII 圖目錄 XI 表目錄 XIII 第一章 前言 1 1.1研究動機 1 1.2研究目的 3 第二章 文獻回顧 5 2.1抗生素抗性基因 5 2.1.1抗生素抗性基因來源與機制 5 2.1.2環境中的抗生素抗性基因 7 2.2 重金屬 9 2.2.1 底泥重金屬 9 2.2.2 重金屬污染影響底泥中微生物生態 12 2.2.3 重金屬影響抗生素抗性基因之分佈與傳播 13 2.2.3.1 共選擇機制 13 2.2.3.2 可動遺傳因子-Ⅰ類整合子(Class 1 Integrons) 15 2.2.3.3 環境中的共選擇機制 16 2.3 臺灣河口重金屬污染 17 2.4 次世代定序應用於微生物族群結構分析 18 第三章 材料與方法 19 3.1 實驗架構 19 3.2 現地採樣 20 3.2.1 新竹香山濕地 20 3.2.2 西南沿海 20 3.2.3 採樣點位資訊 20 3.2.4 採樣方法 23 3.3 底泥物化分析 23 3.3.1 粒徑分佈 23 3.3.2 總有機質 24 3.3.3 重金屬 24 3.4 分子生物實驗 25 3.4.1 底泥核酸萃取 25 3.4.2 抗生素抗性基因標準品製備 26 3.4.2.1 動物排泄物之核酸萃取 26 3.4.2.2 抗生素抗性基因之擴增 26 3.4.2.3 TA cloning 30 3.4.2.4 質體萃取 31 3.4.3 即時聚合酶連鎖反應定量分析抗生素抗性基因 31 3.5 次世代定序 34 3.5.1 聚合酶連鎖反應(PCR)及定序文庫製備(Library preparation) 34 3.5.2 Illumina Miseq定序 35 3.5.3 序列整理(Read processing) 35 3.5.4 操作分類單元(OTU)分析及菌種比對 35 3.6 統計分析 37 第四章 結果與討論 39 4.1 底泥物化性質 39 4.2 抗生素抗性基因及Ⅰ類整合子(CLASS 1 INTEGRONS)之分佈 42 4.3 Ⅰ類整合子與抗生素抗性基因之相關性 48 4.4 抗生素抗性基因、Ⅰ類整合子與環境變量之相關性 49 4.5 共選擇之風險 53 4.6 次世代定序數據 54 4.6.1 菌科組成百分比 54 4.6.2 樣點間微生物族群差異 57 4.6.3 微生物族群(菌科與菌屬)與環境變量之相關性 60 4.6.4 重金屬顯著正相關之菌群 70 4.6.5 抗生素抗性基因的潛在宿主 73 第五章 結論與建議 77 5.1 結論 77 5.2 建議 78 參考文獻 79 附錄 93 附錄1 QPCR標準曲線 93 附錄2 QPCR MELTING CURVE 97 附錄3 各重金屬之SPEARMAN相關性分析 101 附錄4 重金屬與底泥基本性質之SPEARMAN相關性分析 101 | |
dc.language.iso | zh-TW | |
dc.title | 重金屬與菌群結構及抗生素抗性基因於河口底泥的分佈之關聯 | zh_TW |
dc.title | Impact of heavy metal to microbial community antibiotic resistance genes in estuary sediment | en |
dc.type | Thesis | |
dc.date.schoolyear | 108-2 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 江殷儒(Yin-Ru Chiang),于昌平(Chang-Ping Yu),林居慶(Chu-Ching Lin) | |
dc.subject.keyword | 河口底泥,重金屬,菌群結構,抗生素抗性基因,共選擇,水平基因轉移, | zh_TW |
dc.subject.keyword | estuary sediments,heavy metals,microbial community structure,antibiotic resistance genes,co-selection,horizontal gene transfer, | en |
dc.relation.page | 101 | |
dc.identifier.doi | 10.6342/NTU202002774 | |
dc.rights.note | 未授權 | |
dc.date.accepted | 2020-08-11 | |
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 |
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