吸附汞之三價鐵礦於生物還原溶解過程中元素汞的生成與移動潛勢

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研究生：	王詩芸 Shih-Yun Wang
論文名稱：	吸附汞之三價鐵礦於生物還原溶解過程中元素汞的生成與移動潛勢 Reductive dissolution of mercury-bearing iron(III) (oxyhydr)oxides by dissimilatory iron-reducing bacteria and the potential to mobilize mercury in its elemental form
指導教授：	林居慶
口試委員:
學位類別：	碩士 Master
系所名稱：	工學院 - 環境工程研究所 Graduate Institute of Environmental Engineering
論文出版年：	2016
畢業學年度：	104
語文別：	中文
論文頁數：	105
中文關鍵詞：	異化性鐵還原、地下含水層、元素汞生成、汞移動潛勢
外文關鍵詞：	dissimilatory iron reducing bacteria, aquifers, formation of elemental mercury, mercury mobility
相關次數：	點閱：12 下載：0
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汞是毒性最強的元素之一，當地下水遭受汞污染時將導致此水資源無法被適
當利用。在許多案例中，污染場址並未發現到有明顯外來點源的存在，且水質監
測的結果常顯示(1)受污染的地下水含有較高濃度的有機碳，以及(2)水中的總汞、
甚至元素汞濃度的增加與溶解鐵濃度的增加具有正相關等特性。雖然幾種可能的
生地化機制已被提出用以解釋及推估污染事件背後的成因，但這些機制目前皆未
被證實，包括「當含水層處於以鐵還原為主要的最終電子接受的狀態時，汞化性
與移動性的轉變極有可能與系統中的異化性鐵還原菌之生長及代謝有關」這樣的
論點，其推論主要是基於過往的研究已證實鐵還原菌本身以及因這類微生物生長
代謝而衍生的二次礦物可將汞直接還原。然而，文獻上所得的這些汞還原作用其
實驗設計皆是針對水溶液中的汞進行探討，無法貼近於地下水污染案例中汞物種
真實的分布情況。有鑑於此，本研究藉由模擬的方式以可進行胞外呼吸作用的鐵
還原菌 Shewanella oneidensis MR-1 為模式生物，環境中最常見的三價鐵礦hematite (赤鐵礦)與 goethite (黃鐵礦)為模式礦物進行厭氧縮模試驗，待 Hg(II)與hematite 或 goethite 於試驗培養液中達吸附平衡後加入 MR-1，並隨時間分析系統中的溶解汞、元素汞與總亞鐵的含量變化，以觀測地下含水層中當異化性鐵還原菌受刺激生長後二次鐵礦的形成及汞的氧化還原轉換與移動潛勢。試驗與模擬的結果顯示，Hg(II)與 Fe(III)鐵礦可能因靜電吸引作用的關係而迅速地達到吸附平衡，且此平衡在添加 MR-1 菌株後並未受到劇烈破壞。此外，汞的還原程度雖然隨著 MR-1 對不同鐵礦的使用而有所差異，但不論是在含有 hematite 或 goethite的系統，元素汞的生成皆可對應細胞的生長代謝，且過程中所檢測到的溶解汞濃度的變化皆不甚明顯。這些結果支持原本的假說，即「地下飽和含水層中原吸附/鑲嵌於鐵礦的 Hg(II)在受到鐵還原菌生長代謝的影響下，將因生源性 Fe(II)的生成而易被還原成 Hg(0)，使得整體 Hg 於地下水環境中的移動性提高，進而促成汞的污染擴散」。

The groundwater contamination with mercury (Hg) is an increasing problem worldwide, and since Hg is highly toxic, contamination may render this water resource unsuitable for intented use. In many cases, the external pollution sources are not well defined, and samples taken from monitoring wells are oftentimes characterized with (i) elevated levels of organic carbon and (ii) a positive correlation between concentrations of total Hg or elemental Hg and dissolved iron (Fe). On the basis of these
observations, possible mechanisms of the biogeochemical processes that underlie the contamination scenarios have been proposed, including the most acceptable one that
postulates “growth and associated metabolisms of indigenous iron-reducers may have been the primary cause for the alternation of Hg speciation and mobility in the aquifer”.
However, such hypothesis is completely derived from results of the studies that only investigated redox transformations of Hg in the aqueous phase of a heterogeneous
system, which may not be representative of the real situation encountered in the aquifer, as Hg is considered originating from saturated Hg-bearing sediment (i.e., the
solid phase). To validate this hypothesis, laboratory microcosm experiments were conducted to simulate processes of microbially-induced reductive dissolution of Fe(III)
minerals by equilibrating Hg(II) adsorption onto synthetic hematite and goethite (two of the most commonly found iron oxyhydroxides in the environment used as model minerals) prior to the addition and growth of the model bacterium, Shewanella oneidensis MR-1 (an iron reducer). Hg(0), dissolved Hg(II) and total Fe(II) were measured periodically over the course of the experiment to monitor the trends of phase distribution, redox transformation and thus mobility of Hg resulted from microbial
growth coupled with transit of secondary iron mineral formation. It was observed that equilibrium of Hg(II) adsorption onto Fe(III) minerals was reached rapidly presumably due to strong electrostatic interactions, and was not significantly disturbed after the
spike of bacterial cells. In addition, formation of Hg(0) corresponding to the growth of cells, as well as little detection of dissolved Hg(II) were observed. These results support the original hypothesis and indicate that Hg(II) deposited in the sedimentary zone would be reduced and released as Hg(0) on account of biogenic Fe(II) produced
by iron-reducing bacteria and therefore the entire process potentially promotes the movement of Hg in the aquifer environment.

目錄 
 
中文摘要 ......................................................................................................................... I 
Abstract ....................................................................................................................... III 
目錄 .............................................................................................................................. ..V 
圖目錄 ...................................................................................................................... .VIII 
表目錄 ........................................................................................................................ . XI     
第一章  前言 ................................................................................................................... 1 
1.1  研究背景 ........................................................................................................... 1 
1.2  研究目的 ........................................................................................................... 5 
第二章  文獻回顧 ........................................................................................................... 6 
2.1  汞的型態與特性 ............................................................................................... 6 
2.2  環境中汞的排放源 ........................................................................................... 7 
2.3  汞的毒性與相關法規 ....................................................................................... 9 
2.4  汞於環境中的流佈情形 ................................................................................. 11 
2.5  地下水汞污染案例 ......................................................................................... 13 
2.6  汞在土壤及地下水的生地化反應 ................................................................. 16 
2.7  汞在地下環境的沉澱錯合與吸附反應 ......................................................... 16 
2.8  汞在地下環境的非生物性還原反應 ............................................................. 18 
2.9  異化性鐵還原菌 ............................................................................................. 21 
第三章  實驗材料、方法與設備 ................................................................................. 23 
3.1  實驗架構 ......................................................................................................... 23 
3.2  實驗藥品與試劑 ............................................................................................. 24 
3.2.1  實驗試劑 ........................................................................................................ 24 
3.2.2  試驗培養液成分與製備 ................................................................................ 26 
3.3  實驗設備與儀器 ............................................................................................. 32 
3.3.1  實驗設備 ........................................................................................................ 32 
3.3.2  分析用儀器 .................................................................................................... 34 
3.4  化學物種組成模擬軟體 ................................................................................. 34 
3.5  氧化鐵礦製備與鑑定 ..................................................................................... 35 
3.5.1  氧化鐵礦合成 ................................................................................................ 35 
3.5.2  氧化鐵礦定性分析 ........................................................................................ 36 
3.6  實驗用之模式菌種 ......................................................................................... 37 
3.7  氧化鐵礦吸附汞離子實驗 ............................................................................. 37 
3.7.1  比較以去離子水及分析培養液進行汞吸附實驗 ........................................ 38 
3.7.2  添加死菌吸附平衡測試 ................................................................................ 39 
3.8 Shewanella oneidensis MR-1 還原鐵礦測試 .................................................. 40 
3.9  異化性鐵還原微生物縮模實驗 ..................................................................... 41 
3.10  分析方法 ....................................................................................................... 42 
3.10.1  汞分析 .......................................................................................................... 42 
3.10.2  鐵分析 .......................................................................................................... 44 
第四章  結果與討論 ..................................................................................................... 46 
4.1  鐵礦合成及基本特性分析 ............................................................................. 46 
4.1.1  鐵礦形貌 ........................................................................................................ 46 
4.1.2  晶型結構鑑定 ................................................................................................ 48 
4.1.3  鐵礦表面電位分析 ........................................................................................ 49 
4.1.4  比表面積及粒徑分析 .............................................................................. 51 
4.2  汞回收率試驗 ................................................................................................. 52 
4.3  合成氧化鐵礦吸附實驗 ................................................................................. 54 
4.3.1  比較不同水溶液條件下汞的吸附實驗 ........................................................ 60 
4.3.2  汞脫附實驗 .................................................................................................... 62 
4.4 Shewanella oneidensis MR-1 還原鐵礦測試 .................................................. 63 
4.4.1 Shewanella oneidensis MR-1 生長曲線 ......................................................... 63 
4.4.2 Shewanella oneidensis MR-1 還原鐵礦測試 ................................................. 65 
4.5  異化性鐵還原微生物縮模實驗 ..................................................................... 67 
4.5.1  實驗小結 ........................................................................................................ 74 
4.5.2  環境意義 ........................................................................................................ 74 
第五章  結論與建議 ..................................................................................................... 77 
5.1  結論 ................................................................................................................. 77 
5.2  建議 ................................................................................................................. 79 
參考文獻 ....................................................................................................................... 80 
附錄一 ........................................................................................................................... 90 
圖目錄 
 
圖 2-1  自然環境中汞的來源與生地化循環 ................................................................. 7 
圖 2-2  每年全球主要人為排放源之汞排放量 ............................................................. 8 
圖 2-3  不同的 Fe(II)/Fe(III)組合下的氧化還原潛勢(E h ) .......................................... 20 
圖 3-1  吹氣捕捉(purge & trap)系統 ............................................................................ 33 
圖 3-2  吹氣捕捉(Purge and trap )系統設置圖 ............................................................ 33 
圖 3-3  以 Ferrozine 方法定量反應瓶產生之亞鐵 ..................................................... 45 
圖 3-4  分光光度計分析 Fe(II)之檢量線 .................................................................... 45 
圖 4-1  實驗自行製備的鐵礦保存於無氧水中，左為黃鐵礦(goethite)、右為赤鐵礦
(hematite)。 ................................................................................................... 47 
圖 4-2  文獻中氧化鐵礦物所呈現的顏色 ................................................................... 47 
圖 4-3    Hematite 與 Goethite  之 XRD 圖譜 .............................................................. 48 
圖 4-4  以 1 mM MOPS 作為緩衝溶液，hematite 在不同 pH 下的界達電位值。
hematite 零電位點約 7.7。 .......................................................................... 50 
圖 4-5  以 1 mM MOPS 作為緩衝溶液，goethite 在不同 pH 下的界達電位值。goethite
零電位點約 7.81。 ....................................................................................... 50 
圖 4-6  總汞回收率試驗。添加至反應瓶中的 Hg(II)質量為 4 ng。白色條柱代表捕
捉瓶捕捉到的 Hg(0)質量，深灰色條柱代表於吹氣捕捉後反應瓶剩餘的
Hg(II)質量。SnCl 2  spike  為添加氯化亞錫作為還原劑。 ........................ 53 
圖 4-7  兩種鐵礦與  Hg(II)在不同吸附時間下，系統中溶解汞的濃度變化 .......... 56 
圖 4-8  模擬汞吸附至鐵及鋁氧化物表面(假設在沒有汞還原的條件下) ................ 57 
圖 4-9 Hematite 在培養液中不同 pH 下的界達電位值。hematite 零電位點小於 5。
 58 
圖 4-10 Goethite 在培養液中不同 pH 下的界達電位值。goethite 零電位點小於 5。
 58
圖 4-11  比較分別以去離子水及分析培養液進行汞的吸附實驗於反應時間 20 小時
後，Goethite  與 Hematite 反應瓶中汞的濃度 ........................................... 61 
圖 4-12  比較不添加與添加死菌至汞與鐵礦達平衡吸附系統並靜置隔夜, ............ 63 
圖 4-13 S. oneidensis MR-1 培養於 30℃、pH 7 的液態 defind medium 時，測量細
胞於試管中之吸光值隨著時間變化之生長曲線 ....................................... 64 
圖 4-14 S. oneidensis MR-1 在不同鐵礦培養下，於特定時間點採樣分析反應瓶所
生成的總亞鐵濃度。 ................................................................................... 66 
圖 4-15 S. oneidensis MR-1 培養於 goethite 與汞達平衡的系統，於特定時間點採樣
分析反應瓶內生成的總亞鐵濃度，其中  control  為不添加菌株之控制組、
live cells 為實驗組、以及  heat- killed cells  為死菌對照組。 ................. 69 
圖 4-16 S. oneidensis MR-1 培養於 hematite 與汞達平衡的系統，於特定時間點採樣
分析反應瓶內生成的總亞鐵濃度，其中  control  為不添加菌株之控制組、
live cells 為實驗組、以及  heat- killed cells  為死菌對照組。 ................. 69 
圖 4-17 S. oneidensis MR-1 培養於 goethite 與汞達平衡的系統，於特定時間點採樣
分析反應瓶內溶解汞的質量。 ................................................................... 70 
圖 4-18 S. oneidensis MR-1 培養於 hematite 與汞達平衡的系統，於特定時間點採樣
分析反應瓶內溶解汞的質量。 ................................................................... 70 
圖 4-19 S. oneidensis MR-1 培養於 goethite 與汞達平衡的系統，於特定時間點採樣
分析反應瓶內  Hg(0)的質量。.................................................................... 72 
圖 4-20 S. oneidensis MR-1 培養於 hematite 與汞達平衡的系統，於特定時間點採樣
分析反應瓶內  Hg(0)的質量。.................................................................... 72 
表目錄 
 
表 2-1  國內汞公告管制標準 ...................................................................................... 10 
表 3-1  維持 S. oneidensis MR-1 生長，複合培養基(complex medium)組成 .......... 27 
表 3-2  維持 S. oneidensis MR-1 生長，確定培養液(defind medium)組成 ............. 28 
表 3-3  進行吸附實驗及生物縮模試驗所用的(Assay medium)組成(pH = 7) .......... 29 
表 3-4 Trace elements solution 之組成分 .................................................................... 30 
表 3-5 Vitamin solution (Concentrated 10X)之組成份 ............................................... 31 
表 3-6  氧化鐵礦吸附汞之實驗參數 .......................................................................... 38 
表 3-7  以去離子水及分析培養液進行汞吸附實驗參數 .......................................... 39 
表 3-8  吸附實驗參數-添加死菌試驗 ......................................................................... 40 
表 4-1  與其他文獻之合成氧化鐵 pH pzc 值比較 ........................................................ 49 
表 4-2  合成三價氧化鐵礦 Goethite 與 Hematite 比表面積及粒徑分布分析 ......... 51 
表 4-3  以 Visual MINTEQ 模式模擬汞於 assay medium 中的物種濃度分布， .... 59 
表 4-4  以 Visual MINTEQ 模式模擬汞於 distilled water 中的物種濃度分布， .... 61 
                                

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