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姓名	柏貫中(Kuan Chung Pao)  查詢紙本館藏	畢業系所	環境工程研究所
論文名稱	蘭陽平原地下水水溶氣(甲烷)厭氧氧化作用對含水層砷釋出之影響 (Effect of in situ anaerobic methane oxidation on the mobilization of arsenic)
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摘要(中)	地下水是全世界重要的水資源，含高濃度砷的地下水對仰賴此水資源的民眾造成莫大的健康威脅，為全球公共衛生的重大議題。台灣蘭陽平原地下水具有高濃度的砷，20 % 的水井砷濃度超過 WHO 所訂定的地下水砷濃度建議值 10 ppb，當地居民也非常仰賴地下水資源，因此含砷地下水為居民的潛在健康危害。宜蘭縣冬山鄉武淵國小一帶居民有使用「水溶氣」的紀錄，該地水溶氣主要源自於古地層所蘊含之有機質因厭氧消化所產生的甲烷。國內學者過去在蘭陽平原所做的調查，雖然呈現出當地地下水的高砷濃度很有可能與水溶氣的存在有所關連，但卻仍無直接、實質的證據。有鑒於近年來鐵還原狀態下的厭氧甲烷氧化已引起注意，因此本研究在宜蘭縣武淵地區的水井採樣，除監測現地的生地化參數外，也將井壁的沉積物帶回實驗室進行甲烷添加的厭氧培養試驗，監測 Fe(II)、總砷、As(III)、甲烷濃度變化，並搭配相關的功能性基因的定性定量分析，以及菌種鑑定，以驗證蘭陽平原的水溶氣厭氧氧化作用是否確實為當地地下水高濃度砷的主要驅動力。調查的結果顯示當地地下水確實有砷污染的狀況，且以毒性較高的As(III) 為主要型態，並具有相當含量的 Fe(II)，代表現地環境已為厭氧，除表明有高機率發生鐵還原作用外，也說明該環境適宜厭氧甲烷利用菌群的生存。而菌種鑑定的結果也的確呼應可進行厭氧氧化甲烷作用、並還原Fe(III) 的菌群確實存在。培養試驗的結果顯示非生物與生物性作用都可能共同成為當地砷釋出的驅動力，像是 HCO3- 與砷競爭鐵(氫)氧化物表面的吸附點位，使得砷釋放到地下水中，並且隨著培養實驗進行，實驗組(添加甲烷)與控制組(無額外添加甲烷)皆有鐵還原與砷釋出現象，表示甲烷與其他現地有機物皆能成為當地鐵還原菌的電子供給者以發生鐵還原作用並使砷釋出。然而，培養結果卻觀察到 As(III) 的生成並非微生物厭氧氧化甲烷造成的，推測主要為地下水中的 Fe(II) 將 As(V) 還原所致。本篇研究為第一個利用蘭陽平原地下水中的微生物進行培養實驗並證明生物性的鐵還原作用是造成當地砷釋出的主因，並且甲烷能以電子供給者的角色驅動砷釋出。
摘要(英)	High arsenic (As) concentration in groundwater is a worldwide public health issue, and millions of peoples’ health are threatened. Groundwater in Lanyang plain of Taiwan is contaminated by high content of As. Residents rely heavily on groundwater, so As-containing groundwater poses potential health hazards to local people. In most case, the liberation of As from sediment are caused by microbial reductive dissolution of iron oxide hydroxide, and organic carbon would be used as electron donor. There is plenty of CH4 emerging from wells in Wuyuan township of Lanyang plain, so the hypothesis of this study is CH4 can serve as electron donor for anaerobic menthanotrophs and triggering As(V) or As(III) mobilizing. Biogeochemical parameters and microbial community of the wells in Wuyuan township were analyzed for gaining background data, and sediments in the wells were collected for the anaerobic CH4 adding incubations. The main findings are three. First, the competitive sorption of As and HCO3- to iron oxide hydroxide can trigger As released by abiotic reaction. Second, CH4 and non-CH4 in situ organic can carbon function as electron donor for iron-reducing bacteria and cause As liberate into groundwater by reductive dissolution of iron oxide hydroxide. Third, As(III) generating is most likely because of As(V) reducing by Fe(II) in abiotic reaction. This is the first study proving that the reason of As contamination in Lanyang plain is caused by biological iron reduction, and CH4 can be the role of electron donor.
關鍵字(中)	★ 砷 ★ 地下水 ★ 蘭陽平原 ★ 水溶氣 ★ 甲烷厭氧氧化	關鍵字(英)	★ arsenic ★ groundwater ★ Lanyang plain ★ methane ★ anaerobic oxidation of methane
論文目次	摘要 I Abstract II 誌謝 III 第一章 前言 1 1.1 研究緣起與背景 1 1.1.1 砷對人體的健康危害、暴露途徑及台灣蘭陽平原地下水使用現況 1 1.1.2 砷在水環境中的型態 2 1.1.3 地下水砷污染機制 3 1.1.4 蘭陽平原地下水地化環境簡介與砷污染概況 4 1.1.5 甲烷循還與地下水砷污染之關聯 6 1.1.6 蘭陽平原地下水水溶氣(甲烷) 7 1.2 研究目的 9 第二章 研究方法 11 2.1 研究架構 11 2.2 採樣地點介紹 12 2.3 地下水地質化學參數分析 13 2.4 化學物種組成軟體模擬 19 2.5 水井生物膜採樣 20 2.6 反轉錄定量聚合酶連鎖反應(Reverse transcription-quantitative polymerase chain reaction, RT-qPCR) 21 2.7 次世代定序 25 2.8 培養實驗設計 25 2.9 現地有機物微生物利用實驗 30 第三章 結果與討論 31 3.1 地下水生地化參數分析結果 31 3.1.1 地下水地質化學參數分析結果 31 3.1.2 PHREEQC 模擬結果 36 3.1.3 地下水功能性基因分析 38 3.1.4 NGS 分析結果 40 3.2 培養實驗結果 46 3.2.1甲烷消耗率 46 3.2.2 總 Fe(II) 生成 49 3.2.3水溶液中 Fe(II) 生成 53 3.2.4 水溶液中總砷釋出 55 3.2.5 As(III) 釋出 59 3.2.6 DOC 消耗 62 3.2.7 RT-qPCR分析結果 64 3.2.8 現地有機物微生物利用實驗 86 3.3 環境意義 89 第四章 結論與建議 91 4.1 結論 91 4.2 建議 93 第五章 參考文獻 95 口試委員問題與回答 106 圖目錄 圖 1.1.1 蘭陽平原武淵岩芯影像(中央地質調查所) 9 圖 2.1.1 研究架構 11 圖 2.2.1 採樣水井分布 12 圖 2.2.2 採樣水井照片 (a) WR (b) WY 13 圖 2.3.1 陰離子交換樹脂分離砷物種示意圖 14 圖 2.3.2 甲烷採樣裝置 16 圖 2.5.1 水井沉積物採樣 (a)實際採樣照片 (b)羊毛絨收集沉積物 (c)厭氧缸 21 圖 2.7.1 NGS 流程示意圖 25 圖 2.8.1 自行製備之 ferrihydrite 外觀 27 圖 2.8.2 自行備製之 ferrihydrite X-射線繞射分析 27 圖 2.8.3 培養實驗架設示意圖 29 圖 3.1.1 二採樣點 NGS 分析稀釋曲線 41 圖 3.1.2 二採樣點 NGS 分析 Beta diversity 箱型圖 42 圖 3.1.3 二採樣點微生物(科)相對豐富度 44 圖 3.1.4 二採樣點微生物(屬)豐度聚類熱圖 45 圖 3.2.1 WR 採樣點培養實驗甲烷消耗率 47 圖 3.2.2 WY 採樣點培養實驗甲烷消耗率 48 圖 3.2.3 WR 採樣點培養實驗總 Fe(II) 生成 51 圖 3.2.4 WY 採樣點培養實驗總 Fe(II) 生成 52 圖 3.2.5 Magnetite 磁力吸引示意圖 52 圖 3.2.6 WR 採樣點培養實驗水溶液 Fe(II) 生成 54 圖 3.2.7 WY 採樣點培養實驗水溶液 Fe(II) 生成 55 圖 3.2.8 WR 採樣點培養實驗水溶液總砷釋出 57 圖 3.2.10 Goethite 與 lepidocrocite 生成圖 58 圖 3.2.11 WR 採樣點培養實驗水溶液 As(III) 釋出 (a)第14天 (b)第40天 (c)第90天 (d)第105天 61 圖 3.2.12 WY 採樣點培養實驗水溶液 As(III) 釋出 (a)第14天 (b)第40天 (c)第90天 (d)第105天 62 圖 3.2.13 培養實驗 lactate control DOC 消耗趨勢 (a)WR 採樣點 (b)WY 採樣點 63 圖 3.2.14 WR 採樣點 mcrA 基因培養實驗第 80 天相對定量結果 70 圖 3.2.15 WY 採樣點 mcrA 基因培養實驗第 80 天相對定量結果 70 圖 3.2.16 WR 採樣點 pmoA 基因培養實驗第 80 天相對定量結果 71 圖 3.2.17 WY 採樣點 pmoA 基因培養實驗第 80 天相對定量結果 71 圖 3.2.18 WR 採樣點 arrA 基因培養實驗第 80 天相對定量結果 72 圖 3.2.19 WY 採樣點 arrA 基因培養實驗第 80 天相對定量結果 72 圖 3.2.20 WR 採樣點 mcrA 基因培養實驗第 120 天相對定量結果 73 圖 3.2.21 WY 採樣點 mcrA 基因培養實驗第 120 天相對定量結果 73 圖 3.2.22 WR 採樣點 pmoA 基因培養實驗第 120 天相對定量結果 74 圖 3.2.23 WY 採樣點 pmoA 基因培養實驗第 120 天相對定量結果 74 圖 3.2.24 WR 採樣點 arrA 基因培養實驗第 120 天相對定量結果 75 圖 3.2.25 WY 採樣點 arrA 基因培養實驗第 120 天相對定量結果 75 圖 3.2.26 WR 採樣點 with CH4 培養實驗照片 76 圖 3.2.27 WY 採樣點 with CH4 培養實驗照片 77 圖 3.2.28 WR 採樣點 no CH4 培養實驗照片 78 圖 3.2.29 WY 採樣點 no CH4 培養實驗照片 79 圖 3.2.30 WR 採樣點 BES control 培養實驗照片 80 圖 3.2.31 WY 採樣點 BES control 培養實驗照片 81 圖 3.2.32 WR 採樣點 NaN3 control 培養實驗照片 82 圖 3.2.33 WY 採樣點 NaN3 control 培養實驗照片 83 圖 3.2.34 WR 採樣點 lactate control 培養實驗照片 84 圖 3.2.35 WY 採樣點 lactate control 培養實驗照片 85 圖 3.2.36 現地有機物微生物利用實驗(with biofilm) (a)WR 採樣點 (b)WY 採樣點 87 圖 3.2.37 現地有機物微生物利用實驗(no biofilm) (a)WR 採樣點 (b)WY 採樣點 88   表目錄 表2.3.1 (離子)檢量線、LOD 與 LOQ 19 表2.3.2 (元素、DOC、甲烷)檢量線、LOD 與 LOQ 18 表2.7.1 RT-qPCR primer sets 24 表 2.8.1 培養實驗添加物質 28 表 3.1.1 地下水地質化學參數分析 35 表 3.1.2 地下水金屬、類金屬與原素濃度分析 35 表 3.1.3 WR 採樣點 PHREEQC 沉積物組成模擬 37 表 3.1.4 WY 採樣點 PHREEQC 沉積物組成模擬 38 表 3.1.5 地下水化學物種濃度分布 38 表 3.1.6 地下水功能性基因環境背景 Ct 值 39 表 3.1.7 定序資料處理分析 40 表 3.2.1 WR 採樣點培養實驗第 80 天功能性基因 Ct 值 66 表 3.2.2 WY 採樣點培養實驗第 80 天功能性基因 Ct 值 67 表 3.2.3 WR 採樣點培養實驗第 120 天功能性基因 Ct 值 68 表 3.2.4 WY 採樣點培養實驗第 120 天功能性基因 Ct 值 69
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指導教授	林居慶(Chu-Ching Lin)	審核日期	2022-11-15
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