土壤及地下水被石油碳氫化合物所污染已是愈趨普遍且嚴重的問題，此碳氫化合物包括汽油添加劑-甲基第三丁基醚 (methyl tertiary-butyl ether, MTBE)、苯、甲苯、乙苯及二甲苯 [BTEX (benzene, toluene, ethylbenzene和xylenes)] 及三甲基苯 [TMB (1,2,4-trimethylbnezene及1,3,5-trimethylbenzene)] 均會對人體造成危害；而MTBE具有水溶性高、在地下水環境擴散容易及生物分解性較低之特性，苯在厭氧環境中較甲苯、二甲苯及乙苯不易被分解，且已被公告為致癌性物質。由於主要之石油碳氫化合物均能有效由現地微生物分解去除，本研究利用監測式自然衰減 (monitored natural attenuation, MNA) 整治工法，自然衰減效率及機制常為影響環境生態與節能思維之永續整治工法之關鍵因素，而進行本研究場址之整治成效以瞭解自然衰減應用在於土壤及地下水污染場址之可行性及污染物之自然衰減效率評估。本研究以實場操作及實驗室分析數據進行相關探討，監測期間共進行兩年，並以模式模擬實場之碳氫化合物污染團濃度變化及分布評估。本研究選擇一處油污染場址進行自然衰減效率及機制之評估，並評估以監測式自然衰減工法做為研究場址污染團擴散控制及改善地下水中石油碳氫化合物以BTEX為目標污染物並了解整治工法之可行性。研究中以地質化學參數評估、微生物菌相分析及健康危害評估和統計傳輸模式應用 (包括生物降解率、一階衰減率、生物降解容量、風險評估、Mann-Kendall Test、Sustainable Remediation Tool和BIOSCREEN模式之計算及應用) 作為評估效率及機制之方法，模擬結果可有效評估各種整治工法對環境造成的衝擊及影響，以作為決策者進行整治工法篩選及整治效益評估之依據。分析結果顯示，研究場址之石油碳氫化合物(苯) 污染濃度呈下降趨勢。而由場址之溶氧消耗、硝酸鹽減少、亞鐵離子產生、硫酸鹽消耗及二氧化碳與甲烷生成等結果，可證實生物降解作用存在於研究場址，且在污染物之降解過程中扮演關鍵及重要之角色。結果顯示研究場址78%之BTEX由污染源至下游處經自然衰減而去除 (污染源濃度由 6.2 mg/L 下降至 1.2 mg/L 離污染源120 m處)。此外，研究場址之生物降解容量為23 mg/L，此容量高於場址之地下水污染量 (介於6.2 mg/L)，故自然生物降解機制應可達到去除污染物之目的。評估結果亦顯示，ㄧ階衰減速率介於3.1×10-4至5.9×10-4 day-1之間。而由Mann-Kendall Test分析結果顯示，於監測井中污染源之衰減值介於-1.48至-1.74間，顯示污染區之污染物呈現衰減趨勢。由BIOSCREEN模式模擬結果可知，污染團因自然衰減機制之進行，會被控制在離污染源82 m的距離內。由BIOSCREEN之一階衰減模擬結果顯示，被一階生物降解作用所移除的BTEX污染物比例為91%。因此無論是一階衰減反應或瞬間反應，均可有效降解污染物。由健康風險評估模式模擬污染物質苯於地下水與空氣中致癌風險值為4.1×10-103及2.0×10-8並未超過國內的健康風險值1.0×10-6，而經由自然衰減污染濃度的下降對附近居民的健康風險危害程度指數呈現下降的狀態。由變性梯度膠體電泳 (denaturing gradient gel electrophoresis, DGGE) 所進行之菌相分析結果顯示，研究場址污染區與背景區監測井之菌相有顯著的變化，由此顯示，當場址受到石油碳氫化合物污染，將會造成場址中之微生物產生變化。持續兩年之定序結果顯示，研究場址中具有降解石油碳氫化合物之微生物包含Alcaligenes sp., Aquincola sp., Beggiatoa sp., Bosea sp., Brachymonas petroleovorans CHX, Comamonas sp., Dechloromonas sp., Gallionellaceae sp., Hydrogenophaga sp., Methylibium sp., Mycobacterium sp., Pseudomonas sp., Rhodoferax sp., Rhodocyclaceae sp., Thalassolituus sp., Thiobacillus sp., Variovorax sp.等。研究顯示自然生物降解發生於本污染場址相關證據由下列幾點顯示: (1) 電子接受者的消耗如(硝酸鹽的減少、水中溶氧及硝酸鹽的消耗)；(2) 副產物的生成如 (亞鐵離子的產生、及二氧化碳與甲烷的生成)；(3) 石油碳氫化合物濃度由污染源沿地下水流向下游地區逐漸減少；(4) 研究場區監控井BTEX的濃度有減少的趨勢；(5) 鹼度的上升；(6) 微生物數量的上升；(7) 在洩漏源高污染之監測井pH值降低；(8) 由菌相分析結果得知污染團的介入確實會造成環境中之菌相產生變化。由上述結果可知，本研究場址具有許多降解石油碳氫化合物之微生物，故利用現地微生物降解本場址污染物應可達到一定的成效。監測結果顯示自然衰減機制於研究場址進行中，並也是造成污染物濃度降低的原因之ㄧ，而污染物之衰減速率亦在合理範圍內。因此監測式自然衰減應可搭配其他技術作為本場址之整治工法選項。

Soil and groundwater at many industrial areas and disposal sites is contaminated by hydrocarbons that were released into the environment. Anthropogenic hydrocarbon contamination of soil is a global issue throughout the industrialised world. The objective of this study was to evaluate the effectiveness and mechanisms of natural attenuation (NA) on the remediation of petroleum-hydrocarbon contaminated groundwater caused by gasoline and diesel fuel leakage. The industrial petroleum hydrocarbons (e.g., methyl tertiary-butyl ether, MTBE and benzene, toluene, ethylbenzene, xylenes, BTEX) are among the most ubiquitous organic compounds found in subsurface contaminated environment. Among the cost-effective approach for the remediation of the petroleum technologies, NA has become an effective alternative to the remediation methods for the in situ treatment of contaminated subsurface environments. Natural attenuation occurs at most polluted sites. Monitored natural attenuation (MNA) works best where the source of pollution has been removed. For instance, buried waste must be dug up and disposed of properly. Or it can be removed using other available cleanup methods. After the source is removed, the natural processes get rid of the small amount of pollution that remains in the soil and groundwater. The main objectives of this research was to examine the possibility of adopting monitored natural attenuation as a remediation technique for the contaminated groundwater aquifer. During the effectiveness investigation, site groundwater samples were collected and analyzed for the variations in geochemical indicators, microbial diversity, dominant bacteria, biodegradation rates, biodegradation capacity, and trend of contaminant degradation. Results show that more than 70% of the total BTEX (benzene, toluene, ethylbenzene, and xylenes) removal (dropped from 6.2 mg/L at the source zone to 1.2 mg/L at a 120-m downgradient line) was observed through the NA processes. The calculated biodegradation capacity (23 mg/L) is much higher than the detected concentrations of benzene (6.2 mg/L) within the most contaminated area. Results from Mann-Kendall test reveal that BTEX concentrations were decreasing in most of the monitor wells range from -1.48 to -1.74 in source area, and the calculated first-order NA rates for BTEX were in the range from 3.1×10-4 to 5.9×10-3 1/d. Results from BIOSCREEN simulation suggest that biodegradation was responsible for 91% of BTEX mass reduction at this site. Results show the calculated cumulative benzene cancer risk for groundwater and air in off site is 4.1×10-103 and 2.0×10-8 which is not exceeds the acceptable risk-based carcinogenic risk 1.0×10-6, because the occurrence of NA at the site, the harmful levels of health risks to the residents could be reduced and thus, the risk index for cancer risk was decreased. Results from polymerase chain reaction (PCR), denaturing gradient gel electrophoresis (DGGE), and nucleotide sequence analysis reveal that the petroleum-hydrocarbon plume caused the bacterial variation in microbial communities, and more than 36 different petroleum-hydrocarbon degrading bacteria under aerobic and anaerobic conditions were observed (including Alcaligenes sp., Aquincola sp., Beggiatoa sp., Bosea sp., Brachymonas petroleovorans CHX, Comamonas sp., Dechloromonas sp., Gallionellaceae sp., Hydrogenophaga sp., Methylibium sp., Mycobacterium sp., Pseudomonas sp., Rhodoferax sp., Rhodocyclaceae sp., Thalassolituus sp., Thiobacillus sp., Variovorax sp.) Result indicating that BTEX could be biodegraded by the intrinsic bacterial consortium. Results also reveal that the occurrence of intrinsic biodegradation was the major cause of the limited spreading of the BETX plume and decreased contaminant removal through the plume cross section along the transport path. The observed natural attenuation processes there is strong circumstantial evidence within the plume included the following: (1) depletion of electron acceptors (e.g., dissolved oxygen, sulfate), (2) production of the degradation products (e.g., ferrous iron, sulfide, methane, CO2), (3) decreased BTEX concentrations along the transport path, (4) decreased trend of BTEX concentrations in monitor wells, (5) increased alkalinity, (6) increased microbial populations, (7) decreased pH in the spill source area, and (8) preferential removal of certain BTEX components along the transport path. Results indicate that NA can effectively contain the plume, and intrinsic biodegradation mechanisms were the major causes for contaminant removal. Results will be useful in applying NA as an alternative option for the management of petroleum-hydrocarbon contaminated sites.

Acknowledgement I
Abstract (Chinese) V
Abstract VII
Contents IX
List of Tables XIII
List of Figures XV
CHAPTER 1 Introduction 1
1.1 Motivations 2
1.2 Objectives of research 3
CHAPTER 2 Literature Review 5
2.1 Groundwater world use and contamination 6
2.2 Nonaqueous phase liquids (NAPLs) 11
2.3 Soils pollution of petroleum hydrocarbons 15
2.4 Characteristic of petroleum hydrocarbons 16
2.5 Remediation/Cleanup technologies for contaminated groundwater 24
2.6 Physical treatment technologies 26
2.6.1 Pump-and-treat technology 26
2.6.2 Soil vapor extraction and air sparging 27
2.7 Chemical oxidation remediation technologies 29
2.7.1 Ozone application in groundwater remediation 34
2.7.2 Potassium permanganate in groundwater remediation 35
2.7.3 Persulfate application in groundwater remediation 35
2.8 Permeable reactive barrier systems 38
2.9 Bioremediation 46
2.9.1 Introduction of groundwater bioremediation 46
2.9.2 Subsurface aerobic bioremediation 51
2.9.3 Subsurface anoxic/anaerobic biodegradation 56
2.10 Monitored natural attenuation 59
2.10.1 Monitored natural attenuation of contaminated groundwater 59
2.10.2 Contaminated sites management 60
2.11 Current research and future trends in bioremediation 67
CHAPTER 3 Materials and Methods 69
3.1 Study site and monitoring network installation 70
3.2 Sampling procedures 73
3.3 Sample analyses 75
3.4 Mann-Kendall test 79
3.5 Calculation of natural attenuation rates 79
3.6 Biodegradation capacity calculation 80
3.7 BIOSCREEN 81
3.8 Sustainable remediation tool (SRT) 83
3.8.1 Tier 1 system 86
3.8.2 Tier 2 system 87
3.9 Risk-based corrective action (RBCA) assessment 90
3.10 DGGE and PCR 92
3.10.1 Deoxyribonucleic acid (DNA) extraction 92
3.10.2 Ribonucleic acid (RNA) extraction 93
3.10.3 Denaturing gradient gel electrophoresis (DGGE) 94
CHAPTER 4 Result and Discussion 97
4.1 Field investigation 98
4.1.1 Groundwater analyses 98
4.2 Natural attenuation rate calculation along the BTEX plume 115
4.2.1 Calculation of NA rate 115
4.2.2 Biodegradation capacity 116
4.2.3 MK statistic value (S) calculation 117
4.2.4 Variations in BTEX and TMB concentrations in cross-section 117
4.3 Evaluation and prediction of MNA modeling scheme 119
4.3.1 Analysis of BIOSCREEN 119
4.3.2 Analysis of sustainable remediation tool 121
4.3.3 Human health risk assessment 127
4.4 Evaluation of microbial diversity and dominant microorganisms 131
4.4.1 Change of microbial community structures 131
4.4.2 Identification of the BTEX-degrading bacteria 132

CHAPTER 5 Conclusions and Recommendations 145
5.1 Conclusions 146
5.2 Recommendations 148
CHAPTER 6 References 149

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