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研究生:劉瑋晨
研究生(外文):Wei-chen Liu
論文名稱:以基因分析法評估三氯乙烯污染地下水之微生物整治成效
論文名稱(外文):Use of gene analysis to evaluate the groundwater microbial bioremediation processes of a TCE-contaminated site
指導教授:高志明高志明引用關係劉仲康劉仲康引用關係
指導教授(外文):Jimmy C. M. KaoJong-Kang Liu
學位類別:碩士
校院名稱:國立中山大學
系所名稱:生物科學系研究所
學門:生命科學學門
學類:生物學類
論文種類:學術論文
論文出版年:2009
畢業學年度:97
語文別:中文
論文頁數:95
中文關鍵詞:還原脫氯作用共代謝作用三氯乙烯
外文關鍵詞:reductive dechlorinationtrichloroethylenecometabolism
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三氯乙烯是具揮發性的含氯有機溶劑,在工業上常作為清潔劑清除油脂,但處理不當便造成土壤及地下水污染,可用物理、化學及生物的方法整治受污染的地區。物理方法只是三氯乙烯液相、汽相的轉移,而化學方法則要考慮氧化劑活性及移動性還有遺留下來的產物的問題。生物降解的方法可避免產生有毒產物,但要營造適合微生物降解三氯乙烯的最適條件,本研究以高雄某污染場址進行生物整治。微生物在好氧環境以共代謝作用降解三氯乙烯,在添加糖蜜及營養鹽後,誘導現地降解微生物的生長並共代謝污染物。整治後降解三氯乙烯相關菌群(type Ⅱ methanotrophs)及降解酵素(toluene monooxygenase、toluene dioxygenase、particulate methane monooxygenase)之基因出現頻率皆有增加的趨勢。三氯乙烯濃度在第180天降至管制標準(0.05 mg/L)以下。在厭氧環境,微生物則以還原脫氯作用降解三氯乙烯,添加糖蜜為電子供給者,促進還原脫氯作用進行,整治後Dehalococcoides菌群及其相關降解基因(vcrA)出現頻率皆有增加,而三氯乙烯濃度亦有下降。結果顯示利用微生物降解作用整治三氯乙烯污染場址,無論在好氧或厭氧環境皆可有效降解三氯乙烯。
The industrial solvent trichloroethylene (TCE) is among the most ubiquitous chlorinated compounds found in groundwater pollution. TCE in environment can be removed by physical, chemical and biological procedures. The objective of this pilot-scale study was to apply an enhanced in situ bioremediation technology to remediate TCE-contaminated groundwater. Both aerobic and anaerobic remedial systems were evaluated at a TCE-spill site located in southern Taiwan. In the aerobic test zone, the effectiveness of air, nutrient, and sugarcane molasses injection to enhance the aerobic cometabolism on TCE degradation was evaluated. In the anaerobic test zone, the effectiveness of nutrient and sugarcane molasses injection to enhance the anaerobic reductive dechlorination on TCE degradation was also evaluated. Polymerase chain reaction was applied to analyze the gene variation in TCE-microbial degraders during the treatment process. Results from this study indicate that the aerobic TCE-degraders (type Ⅱ methanotrophs) and the gene of degradation enzymes (toluene monooxygenase, toluene dioxygenase, particulate methane monooxygenase) were detected after the treatment process in the aerobic test zone. Moreover, TCE concentration dropped from approximately 0.1 mg/L to below 0.05 mg/L in the aerobic test zone after six months of treatment. In the anaerobic treatment zone, Dehalococcoides (anaerobic TCE-degrader) and the gene of degradation enzyme (vcrA) were detected and a significant drop of TCE concentration was also observed. Results reveal that both the aerobic cometabolism and anaerobic dechlorination are feasible and applicable technologies to clean up TCE contaminated aquifers.
摘要……………………………………………………………………..Ⅰ
壹 前言…………………………………………………………………1
1.1 研究緣起………………………………………………………1
1.2 三氯乙烯特性…………………………………………………3
1.3 三氯乙烯整治方法……………………………………….…...6
1.4 微生物降解三氯乙烯………………………………..…….….8
1.4.1 好氧環境中三氯乙烯的降解…………………..….…...8
1.4.2 厭氧環境中三氯乙烯的降解…………………….……15
1.5 環境監測…………………………………………………...…16
1.6 以分子生物技術探測三氯乙烯降解活性…………………...17
1.7 研究目的……………………………………………………...17
貳 材料與方法…………………………………………………………18
2.1 監測井位置及採樣時間……………………………………...18
2.2 地下水質監測………………………………………………...21
2.3 地下水環境微生物DNA萃取……………………………….22
2.4 PCR……………………………………………………………23
2.5 PCR產物瓊脂膠體電泳檢視…………………………………23
参 結果與討論…………………………………………………………25
3.1 地下水質監測分析………………………………………….25
3.1.1 好氧環境……………………………………………...25
3.1.2 厭氧環境……………………………………………...28
3.2 地下水中三氯乙烯降解基因及菌群分佈……………….....30
3.2.1 好氧環境………………………………………...……30
3.2.2 厭氧環境…………………………………………...…35
肆 結論………………………………………………………………..39
伍 建議…………………………………………………………..……41
參考文獻………………………………………………………...….…42
圖表…………………..………………………………………………..49
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