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請用此 Handle URI 來引用此文件: http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/23174
完整後設資料紀錄
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dc.contributor.advisor謝文陽
dc.contributor.authorHuey-Jiun Chiangen
dc.contributor.author江蕙君zh_TW
dc.date.accessioned2021-06-08T04:45:45Z-
dc.date.copyright2009-08-03
dc.date.issued2009
dc.date.submitted2009-07-31
dc.identifier.citation參考文獻
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dc.identifier.urihttp://tdr.lib.ntu.edu.tw/jspui/handle/123456789/23174-
dc.description.abstract本研究為了瞭解臺灣沿岸海域中抗四環黴素或抗氯黴素之細菌的豐度及其特性,於2007至2008年間,在臺灣的花蓮沿岸、基隆港口、新竹香山濕地、臺南四草外海設置採樣點進行採樣。 將樣本經過序列稀釋後,吸取定量稀釋水樣加入含有四環黴素或氯黴素的PY培養液進行增殖培養並且利用最大可能數法估算異營性細菌及抗藥性細菌在環境中的數量。實驗結果顯示,本研究各採樣地點所測得四環黴素抗性細菌的豐度明顯高於氯黴素抗性細菌的豐度。 將增殖過後的培養液塗盤待菌落生成後,挑選單一菌落並利用連續畫線法將分離株進行分離、純化和保存。各採樣地點共分離出86株對四環黴素或氯黴素具有抗藥性之細菌。再萃取所有分離株的DNA以PCR將各分離株的16S rDNA增幅擴大,以3種限制酶 (DdeI、RsaI和HhaI) 進行限制片段長度多樣性分析,共可分為15種基因型,代表株經16S rDNA定序,並與標準菌株序列比對,可歸類為Acinetobacter, Aeromonas, Alcanivorax, Alteromonas, Corynebacterium, Escherichia, Pseudoalteromonas, Pseudomonas, Serratia, Shewanella與Vibrio等十一屬之菌種,其中Corynebacterium屬於Actinobacteria,其餘十屬皆屬於γ-Proteobacteria。觀察分離株的生長特性、生理特性及菌落的形態特性和親緣之分析結果顯示A25 (Shewanella sp ), D6 (Pseudomonas sp.) 與X4 (Pseudomonas sp.) 三株分離株均可能為新種。
本研究最先發現水生環境中Altermonas, Alcanivorax 與Corynebacterium可能包含具四環黴素抗性基因之細菌;Acinetobacter, Alcanivorax, Altermonas與Serratia可能包含具氯黴素抗性基因之細菌。
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dc.description.tableofcontents目錄
中文摘要 ........................................................................................................................... I
英文摘要 .......................................................................................................................... II
第一章 前言 .......................................................................................................... 1
第二章 材料與方法 .............................................................................................. 5
第一節 培養基 ........................................................................................................ 5
第二節 樣本採集 .................................................................................................... 6
第三節 培養、計數與分離 .................................................................................... 6
第四節 分離株之形態及生理、生化特性初步檢測 ............................................ 6
第五節 分離株之16S rDNA分析 ........................................................................ 11
第三章 結果 ........................................................................................................ 13
第一節 計數 .......................................................................................................... 13
第二節 抗藥性細菌的分離與篩選 ...................................................................... 14
第三節 各基因型代表株 16S rDNA序列分析 .................................................. 20
第四節 各基因代表株之細部生理生化特性分析 .............................................. 22
第四章 討論 ........................................................................................................ 23
第一節 計數 .......................................................................................................... 23
第二節 臺灣沿岸海水中抗藥性細菌之多樣性 .................................................. 24
第三節 各基因型抗藥性細菌之代表株與相近菌種比較 .................................. 26
第四節 臺灣沿岸海域中抗藥性分離株D6與相近菌種的比較 ....................... 31
第五節 臺灣沿岸海域中抗藥性分離株X4與相近菌種的比較 ....................... 33
第六節 臺灣沿岸海域中抗藥性分離株A25與相近菌種的比較 ..................... 33
第七節 臺灣沿岸海域抗藥性細菌之探討 .......................................................... 34
第五章 結論 ........................................................................................................ 36
表目錄 ............................................................................................................................ 43
表一、分離株16S rDNA RFLP分析結果 ........................................................... 43
表二、各採樣地點所分離株之數目、菌屬及編號 ............................................ 44
表三、各群分離株之生理生化特性 .................................................................... 45
表四、各基因型代表株與已知菌種的16S rDNA相似度 ................................. 46
表五、各群基因型代表株之細部生理生化測試 ................................................ 47
表六、各群代表株抗生素感受性測試 ................................................................ 50
表七、D6與相似菌種的比較表 .......................................................................... 51
表八、X4與相似菌種的比較表 .......................................................................... 52
表九、A25與相似菌種的比較表 ........................................................................ 53
圖目錄 ............................................................................................................................ 54
IV
圖一、採樣地點 .................................................................................................... 54
圖二、利用MPN法估算各採樣點可培養異營性細菌數值 ............................. 55
圖三、各採樣樣點抗四環黴素細菌之生菌數及其佔異營細菌生菌數之千分比 ............................................................................................................................... 56
圖四、各採樣樣點氯黴素細菌之生菌數及其佔異營細菌生菌數之千分比 .... 57
圖五、RFLP 分析之DdeI酶切片段電泳圖以及模擬圖 ................................... 58
圖六、RFLP 分析之HhaI酶切片段電泳圖以及模擬圖 ................................... 59
圖七、RFLP 分析之RsaI酶切片段電泳圖以及模擬圖 ................................... 60
圖八、分離菌株D6之穿透式電子顯微鏡圖 ..................................................... 61
圖九、分離菌株X4之穿透式電子顯微鏡圖 ..................................................... 62
圖十、分離菌株A25之穿透式電子顯微鏡圖 ................................................... 63
圖十一、分離株16S rDNA親緣演化樹分析 ..................................................... 64
附錄目 ............................................................................................................................ 65
附錄一、PY 培養基、PY培養液、PY保存培養基成份 ................................. 65
附錄二、 PYG 培養基、PYN培養液、CM培養液成份 ................................ 66
附錄三、MPN表 .................................................................................................. 67
附錄四、抗生素抑制圈大小判定 ........................................................................ 69
附錄五、分離株D6部分16S rDNA序列 .......................................................... 70
dc.language.isozh-TW
dc.subject抗藥性基zh_TW
dc.subject四環黴素zh_TW
dc.subject氯黴素zh_TW
dc.subject海洋環境zh_TW
dc.subject抗生素zh_TW
dc.subjectmarine environmenten
dc.subjectantibioticsen
dc.subjecttetracyclineen
dc.subjectchloramphenicolen
dc.subjectantibiotic-resistant bacteriaen
dc.title臺灣沿岸海域中抗藥性細菌之分離與特性探討zh_TW
dc.titleIsolation and characterization of antibiotic-resistant bacteria from coastal waters of Taiwanen
dc.typeThesis
dc.date.schoolyear97-2
dc.description.degree碩士
dc.contributor.oralexamcommittee蔡珊珊,陳俊堯,林光慧,李宗徽
dc.subject.keyword四環黴素,氯黴素,海洋環境,抗生素,抗藥性基,zh_TW
dc.subject.keywordtetracycline,chloramphenicol,antibiotic-resistant bacteria,marine environment,antibiotics,en
dc.relation.page70
dc.rights.note未授權
dc.date.accepted2009-07-31
dc.contributor.author-college理學院zh_TW
dc.contributor.author-dept海洋研究所zh_TW
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