臺灣博碩士論文加值系統

本研究主要利用實驗室級之釋氧反應牆系統，模擬受BTEX污染地下水之生物復育，並利用PCR-SSCP之分子生物技術，監測系統中微生物之菌群結構變化，及以生物刺激(biostimulation)(添加氧氣釋放物質與氮源)及生物強化(bioaugmentation)(添加BTEX之分解菌株)等方法，探討微生物分解污染物之能力、菌群分佈及總生菌數三者之消長關係。
釋氧物質(ORC)之管柱試驗結果顯示，於固定進流速度230 cm/day下，釋氧率隨CaO2添加量之增加而提升(5 %~30 %)，但當CaO2達30 %～60 %時之釋氧率皆約為0.22 mg O2/day/g-ORC，顯示CaO2添加至一定比例時，與釋氧率已不再成線性之正比關係。且ORC填充量(100 g 與300 g)與管柱進流流速(3.45、20、40 mL/min)對ORC之釋氧率並無顯著影響，而本研究自製之ORC至少可連續長期穩定釋放氧氣約達35天之久。由釋氧反應牆分解BTEX試驗結果顯示：(1)在BTEX進流濃度30 mg/L下，有添加氮源與未添加者，系統對BTEX之處理能力依序為ethylbenzene＞p-xylene＞toluene＞benzene；(2)在含氮源之生物刺激下，釋氧反應牆對BTEX之去除效率高於未含氮源者，且生物強化作用對於系統之去除效率有提升作用；(3)含氮源與未含氮源組在生物強化作用後之穩定期，對BTEX之去除效率分別為52.4 %與38.9 % (benzene)、72.3 %與51.6 % (toluene)、80.2 %與71.4 % (ethylbenzene)及72 %與71 % (p-xylene)；(4)溶氧供給量與釋氧牆之距離成反比關係，因此距離釋氧牆下游15 cm處所監測到之總生菌數約高於30 cm處之100倍，由此研判距離釋氧牆下游5~30 cm之間係為系統主要進行好氧分解BTEX之處；(5)含氮源與未含氮源組之整體生物相變化趨勢類似，但BTEX之去除效率及總生菌數之增加，可推測添加之氮源對微生物之活性具提升作用；(6)綜合BTEX之去除率、COD、溶氧、總生菌數及菌群結構之結果，有助於評估釋氧反應牆進行受BTEX污染之地下水系統生物復育可行性。

The purposes of this study are to evaluate the bioremediation capabilities using two laboratory-scale permeable reactive barriers (PRB) in a BTEX-contaminated groundwater, and to explore the changes in microbial community in the reactor by using PCR-SSCP. The bioremediation capabilities were evaluated by introducing biostimulation (addition of oxygen-releasing materials and nitrogen) and bioaugmentation (addition of BTEX-degrading cultures). A relationship among biodegradation capability, changes in microbial community and total plate count of microorganisms was determined.
Results of oxygen-releasing materials in laboratory column tests indicates that the oxygen release rates increase with the addition of CaO2 and achieve a constant value of 0.22 mg O2-day-1 g-1-ORC while increasing CaO2 to 30 %–60 %, thereby revealing that the linear relationship between oxygen release rates and addition of CaO2 is not consistently observed while keeping increasing CaO2. Moreover, it is found that the total packing amounts of ORC and inlet flowrate had no significant effects on oxygen release rate and the ORC releasing system could consistently release oxygen for at least thirty five days. Results of the permeable reactive barriers show that (1)the removal capability for BTEX decreases in the order of ethylbeneze, p-xylene, toluene, benzene for both nitrogen addition and no nitrogen addition under BTEX concentrations at 30 mg l-1; (2)the removal efficiency of PRB is higher in the nitrogen addition condition for biostimulation comparing with no nitrogen addition condition, and an increased pattern for removal was observed during the bioaugmentation process; (3) the BTEX removals for nitrogen addition and no nitrogen addition are 52.4 % and 38.9 % for benzene, 72.3 % and 51.6 % for toluene, 80.2 % and 71.4 % for ethylbenzene, and 72 % and 71 % for p-xylene; (4)the amount of dissolved oxygen is found to be inversely proportional to the distance from PRB, as evidenced by the average bacteria densities are two orders higher in a location at 15 cm than at 30 cm from the PRB), thereby revealing that the primary aerobic biodegradation zone is in the ranges from 5 to 30 cm downstream of the PRB; (5)the microbial community structure is similar in both the nitrogen addition and in no nitrogen addition conditions, though the removal efficiency of BTEX and the bacteria densities increase, indicating the nitrogen addition stimulates the activity of microorganisms; and (6)determination the relationship among the BTEX removal efficiency, COD, DO, bacteria densities and the microbial community structure provides assistance in evaluating the feasibility of bioremediation using PRB in a BTEX-contaminated groundwater.

目錄
封面內頁
簽名頁
碩博士論文暨電子檔案上網授權書 iii
中文摘要 iv
英文摘要 vi
致謝 viii
目錄 x
圖目錄 xiii
表目錄 xvi
第一章 緒論
1.1 前言 1
1.2 研究目的及內容 2
第二章 文獻回顧
2.1 BTEX特性簡介 4
2.2 芳香族碳氫化合物之生物降解 8
2.1.1 分解芳香族碳氫化合物之微生物 8
2.2.2 BTEX之生化代謝途徑 9
2.3 土壤與地下水污染整治相關技術介紹 13
2.3.1 地下水整治技術之限制 13
2.3.2 國內外地下水污染整治現況 14
2.4 透水性反應牆整治污染之地下水研究 19
2.4.1 透水性反應牆處理技術 19
2.4.2 透水性反應牆應用現況 23
2.4.3 釋氧物質之特性 24
2.4.4 釋氧反應牆之相關研究 25
2.5 生物刺激法與生物強化法於復育整治之應用 34
2.5.1 生物刺激法 34
2.5.2 生物強化法 35
2.6 生物復育過程之菌群結構分析 39
2.6.1 PCR-SSCP分析技術之相關原理與方法 40
2.6.2 聚合酶鏈鎖反應原理 40
2.6.3 單股DNA構形多型性分析 41
第三章 材料與方法
3.1 研究材料與儀器設備 48
3.1.1 菌種來源 48
3.1.2 藥品種類 49
3.1.3 分子生物之儀器設備 53
3.2 研究方法與步驟 54
3.2.1 BTEX批次降解能力評估 54
3.2.2 釋氧物質之管柱試驗 55
3.2.3 釋氧反應牆之組裝 57
3.2.4 釋氧反應牆背景實驗 60
3.2.5 釋氧反應牆分解BTEX實驗 60
3.2.6 釋氧反應牆之去除效率評估方法 64
3.2.7 分子生物技術建立 69
第四章 結果與討論
4.1 批次實驗測試 74
4.2 釋氧物質之管柱試驗 77
4.2.1 ORC配比試驗 77
4.2.2 ORC填充量試驗 80
4.2.3 管柱進流量試驗 81
4.2.4 ORC釋放氧氣時間延遲試驗 82
4.3 釋氧反應牆背景實驗 83
4.3.1 BTEX貫穿試驗 83
4.3.2 ORC之溶氧與氮源監測 84
4.4 釋氧反應牆分解BTEX之實驗結果 86
4.4.1 BTEX去除效率分析 87
4.4.2 水質分析 92
4.4.3 釋氧反應牆系統之菌群分佈 102
第五章 結論與建議
5.1 結論 113
5.2 建議 115
參考文獻 116
附錄一 本研究菌種定序結果 129

圖目錄
圖2.2-1 芳香族碳氫化合物之生化代謝途徑 12
圖2.3-1 現地地下水主要之生物整治技術 18
圖2.3-2 現地地下水生物整治之主要污染物 18
圖2.3-3 受BTEX污染之主要地下水整治方法 18
圖2.4-1 各類型透水性反應示意圖 21
圖3-1 論文整體研究架構圖 47
圖3.2-1 釋氧物質釋氧率之管柱試驗裝置 57
圖3.2-2 釋氧反應牆示意圖 59
圖3.2-3 釋氧反應牆透視圖 59
圖3.2-4 釋氧反應牆分解BTEX之實驗流程圖 63
圖3.2-5 釋氧反應牆之BTEX濃度檢量線 65
圖3.2-6 分子生物技術分析流程圖 69
圖4.1-1 現地混合菌含50 mg/L之NO3--N對BTEX分解情形 75
圖4.1-2 現地混合菌不含NO3--N對BTEX分解情形 75
圖4.1-3 BTEX分解菌含50 mg/L之NO3--N對BTEX分解情形 76
圖4.1-4 BTEX分解菌不含NO3--N對BTEX分解情形 76
圖4.2-1 不同CaO2添加比例之溶氧變化率 78
圖4.2-2 不同CaO2添加比例之單位ORC釋氧率 79
圖4.2-3 不同ORC填充量試驗 80
圖4.2-4 不同管柱流量試驗 81
圖4.2-5 ORC釋放氧氣時間試驗 82
圖4.3-1 BTEX貫穿實驗結果 84
圖4.3-2 釋氧牆之溶氧監測結果 85
圖4.3-3 釋氧牆之NO3--N監測結果 86
圖4.4-1 A組對BTEX之去除效率(含氮源) 89
圖4.4-2 B組對BTEX之去除效率(不含氮源) 90
圖4.4-3 A組對BTEX之平均去除效率(含氮源) 91
圖4.4-4 B組對BTEX之平均去除效率(不含氮源) 91
圖4.4-5 A組ORP變化量(含氮源) 93
圖4.4-6 B組ORP變化量(不含氮源) 93
圖4.4-7 A組pH變化量(含氮源) 94
圖4.4-8 B組pH變化量(不含氮源) 94
圖4.4-9 A組COD變化量(含氮源) 96
圖4.4-10 B組COD變化量(不含氮源) 96
圖4.4-11 A組DO變化量(含氮源) 100
圖4.4-12 B組DO變化量(不含氮源) 100
圖4.4-13 A組總生菌數變化量(含氮源) 101
圖4.4-14 B組總生菌數變化量(不含氮源) 101
圖4.4-15 BTEX分解菌之指紋圖譜 103
圖4.4-16 A組# 1監測井菌群結構間之相對相似度及群叢關係與SSCP指紋圖譜。(a)SSCP指紋圖譜；(b)相對相似度及群叢關係圖 107
圖4.4-17 B組# 1監測井菌群結構間之相對相似度及群叢關係與SSCP指紋圖譜。(a)SSCP指紋圖譜；(b)相對相似度及群叢關係圖 108
圖4.4-18 A組# 3監測井菌群結構間之相對相似度及群叢關係與SSCP指紋圖譜。(a)SSCP指紋圖譜；(b)相對相似度及群叢關係圖 109
圖4.4-19 B組# 3監測井菌群結構間之相對相似度及群叢關係與SSCP指紋圖譜。(a)SSCP指紋圖譜；(b)相對相似度及群叢關係圖 110
圖4.4-20 A組# 4監測井菌群結構間之相對相似度及群叢關係與SSCP指紋圖譜。(a)SSCP指紋圖譜；(b)相對相似度及群叢關係圖 111
圖4.4-21 B組# 4監測井菌群結構間之相對相似度及群叢關係與SSCP指紋圖譜。(a)SSCP指紋圖譜；(b)相對相似度及群叢關係圖 112

表目錄
表2.1-1 BTEX之物化特性 6
表2.1-2 國內目前公告之地下水污染管制標準 7
表2.2-1 微生物代謝基本形式 10
表2.2-2 已知可降解BTEXl之微生物 11
表2.3-1(a) 地下水污染整治技術彙整(1/2) 16
表2.3-2(b) 地下水污染整治技術彙整(2/2) 17
表2.4-1 透水性反應牆對污染物之處理型式 22
表2.4-2(a) 現地以透水性反應牆整治地下水污染現況彙整(1/4) 28
表2.4-3(b) 現地以透水性反應牆整治地下水污染現況彙整(2/4) 29
表2.4-4(c) 現地以透水性反應牆整治地下水污染現況彙整(3/4) 30
表2.4-5(d) 現地以透水性反應牆整治地下水污染現況彙整(4/4) 31
表2.4-6(a) 應用釋氧反應牆整治地下水污染之研究(1/2) 32
表2.4-7(b) 應用釋氧反應牆整治地下水污染之研究(2/2) 33
表2.5-1(a) 國內外生物刺激及強化法之研究(1/2) 37
表2.5-2(b) 國內外生物刺激及強化法之研究(2/2) 38
表2.6-1(a) 國內外分子生物應用技術之研究(1/4) 42
表2.6-2(b) 國內外分子生物應用技術之研究(2/4) 43
表2.6-3(c) 國內外分子生物應用技術之研究(3/4) 44
表2.6-4(d) 國內外分子生物應用技術之研究(4/4) 45
表3.1-1 各菌種之比對結果與特性 49
表3.1-2 碳源藥品清單 49
表3.1-3 模擬地下水之營養鹽配比 50
表3.1-4 釋氧物質之相關藥品及材料 50
表3.1-5 寡核苷酸引子種類 51
表3.1-6 PCR與SSCP相關藥品清單 51
表3.1-7 分子生物技術之相關藥品種類 52
表3.1-8 分子生物技術之儀器設備清單 53
表3.2-1 管柱試驗之實驗項目與操作參數 56
表3.2-2 釋氧反應牆系統規格 58
表3.2-3 石英砂之化學成分及粒徑分佈 59
表3.2-4 釋氧反應牆之操作條件 63
表3.2-5 氣相層析儀之分析條件 66
表3.2-6 水質分析方法或儀器 68
表3.3-7 PCR升溫程式 71
表3.2-8 PCR藥品與比例 72
表4.4-1 A與B組平均去除效率之比較表 92

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