臺灣博碩士論文加值系統

台灣污染場址土壤多以重金屬污染為主。重金屬在土壤中移動慢，無法被生物有效分解，也不易揮發，因此容易透過生物濃縮或生物放大作用，累積於人體中。目前雖已有多種土壤重金屬復育技術，但經濟可行、效率高，又能保持原土壤特性者，乃有待開發。本研究以犀角金龜幼蟲為生物反應器，探討其食入三種土壤常見重金屬銅鋅鎳後的流布，其間並記錄蟲體生長指標，以評估蟲體在不同濃度重金屬暴露下的生理變化，藉以評估以甲蟲復育重金屬污染土壤的可行性與可能移除效率。為此，本研究主要目的如下：(1) 找出目標重金屬對犀角金龜幼蟲之劑量效應、(2) 評估犀角金龜幼蟲對去除污染物之效率、(3) 研究犀角金龜幼蟲期、化蛹期和成蟲期之重金屬質量流布。研究中先在甲蟲飼育材中添加銅、鋅、鎳等重金屬後，觀察甲蟲幼蟲生長狀況，並進一步測試其對添加重金屬去除效率。接著將已暴露於重金屬十週蟲體、最後一次蛻皮及蛹殼及成蟲不同部位，進行金屬含量檢測。最後，根據檢測資料，就重金屬在不同時期流布數據進行分析。
研究結果如下：蟲體在十週飼養期間死亡率方面，只有鋅濃度組幼蟲死亡率在25-50%間，高於空白組(死亡率25%)；在蟲體吃食量方面，前三、四週並無異常，第四週後暴露組吃食量明顯低於空白組，而其中鎳7,200 ppm組蟲體吃食量大幅下降；在體重成長率及基質轉換率方面，前五週蟲體成長與空白組相似，僅鎳7,200 ppm組蟲體明顯偏低；在糞便累積量方面，銅和鋅添加組跟空白組結果雷同，而鎳7,200 ppm組則因為受到進食量減少影響，導致累積糞便量也跟著下降；另，幼蟲累積於其體內的重金屬去除效率介於40-80%，而蟲體內累積重金屬量與暴露濃度呈正比關係，且平均單隻蟲體內累積最高銅、鋅及鎳，分別為125.4、91.4及595.3 mg。由金屬流布數據中得知: 蟲體內重金屬主要累積在腸道內，銅比鋅跟鎳更容易從腸道中擴散至幼蟲體內，而體內銅與鎳含量並不會影響幼蟲化蛹，但鋅含量上升，幼蟲化蛹成功率則隨之下降。化蛹期的蛻皮(shed)及蛹殼(puparium)總重金屬占比為，銅濃度組30-49%，鋅濃度組25-30%，鎳濃度組100%(含蛹體)，且在相同金屬濃度(即1,200 ppm)條件下，脫皮的銅、鋅和鎳含量分別為30.8、6.9和13.6 mg，蛹殼含量則分別為12.7、15.0和48.9 mg (其中鎳組因未羽化，故以整個蛹體量表示)。成蟲期數據顯示，銅金屬分布於成蟲體內較為平均，而鋅金屬分布則主要累積於頭部(head)、腿部(legs)和甲蟲身體(body remains); 而濃度方面，銅組在嘴器(mouthpart)及內翅(membrane wings)有高濃度累積，而鋅組在嘴器、內翅及鞘翅(elytra)中並無金屬累積。此外，本研究最後以土壤/飼育材等於1的條件下進行土壤重金屬移除測試，其間甲蟲生長狀況及所添加銅鋅鎳去除率與無土壤結果相當。故以甲蟲來移除土壤重金屬，應屬可行。

Taiwan’s soil is mainly polluted with heavy metals. In soil heavy metal moves slowly, is neither degradable nor volatile, which allows them to accumulate inside the human body. Thus, certain novel environmentally friendly technology to remove heavy metals in the soil is eagerly needed. In this study, larvae of Oryctes rhinoceros are used as bioreactors to investigate their metabolic behaviors and heavy metal removal efficiency after the heavy metals (i.e., copper, zinc, and nickel) being ingested. Meanwhile, the feasibility of using beetle larvae as bioreactors for soil heavy metal removal was to be evaluated. Thus, the main objectives of this study include: (1) to investigate the dosage effects of each targeted metal to rhinoceros beetle larvae, (2) to assess the soil metal removal efficiency using beetle larvae, and (3) to study the fate of the heavy metals at larva, and pupa/adult stages. Heavy metals including copper, zinc and nickel, were spiked into the beetle’s feed to monitor the larval growth conditions and the fate of heavy metal in larvae. Furthermore,–the larvae at week ten were to be sacrificed along with the shed and puparium during pupation analyzed for metal contents.
The obtained results are as follows: for the larval mortality rate during the first ten weeks, only the zinc groups had higher mortality at 25-50% that was higher than the Blank of 25%, the rest groups had much lower larval mortality rates. For the feed consumption, the heavy metal spiked groups had similar feed consumption as the Blank for the first 3-4 weeks; however, since the fourth week the heavy metal groups had less feed consumption and the Ni-7200-ppm group had even less feed consumed. For the cumulative feces production, Cu- and Zn-group had similar results as the Blank, yet the Ni-7200-ppm group had significantly less amount. Moreover, the larval heavy metal accumulative rates were around 40-80%, and the cumulative amount was proportional to its feed heavy metal content. The highest averaged cumulative copper, zinc and nickel were 125.4, 91.4 and 595.3 mg per larva. The fate results of the heavy metals in larvae showed that all the ingested heavy metals accumulated in guts and copper diffused to the larval body more than zinc and nickel. The accumulated amount of copper or nickel did not affect the larval pupation, yet the more zinc in larva the high failure of larval pupation. During the pupation period, the Cu, Zn and Ni content in the shed were 30-49, 25-30 and 100% (i.e., the whole pupa), respectively. At the same feed heavy metal concentration of 1200 ppm, the averaged copper, zinc and nickel concentrations in the larval shed were 30.8, 6.9 and 13.6 mg, respectively, and in the larval puparium 12.7, 15.0 and 48.9 mg (i.e., the whole pupa), respectively. As for the fate of the heavy metals in adult larvae, the copper was more evenly distributed in the whole adult body, yet the zinc and nickel were mostly accumulated in head, legs and body remains. Concentration-wise, the beetle mouthpart, and membrane wings had high copper contents, yet there had no zinc detected in the beetle mouthpart, and membrane and elytra wings. Finally, when breeding beetle larvae at an equal amount of heavy metal polluted soil and feed mixture, the larval growth data and the heavy metal removal efficiencies were nearly identical, which implied the probability of using the beetle larvae as bioreactors to remove soil heavy metals.

摘要
Abstract
致謝
表目錄
圖目錄
第一章 研究緣起與目的
第二章 文獻回顧
2-1 目標污染物重金屬
2-1-1 重金屬之來源
2-1-2 常見重金屬之危害性
2-2 土壤污染之整治修復技術
2-2-1 土壤污染整治技術分類及比較
2-2-2 土壤重金屬去除技術
2-2-3 微生物復育技術
2-2-4 大型生物復育技術
2-3 兜蟲介紹
2-4 犀角金龜
2-4-1 犀角金龜用於土壤復育案例
2-4-2 重金屬對犀角金龜幼蟲的毒性及其可能流布
2-4-3 重金屬在甲蟲體內流布與追蹤
第三章 研究材料與方法
3-1 研究架構與流程
3-2 最適犀角金龜幼蟲生長環境條件
3-3 實驗藥品與材料
3-4 實驗設備
3-4-1 感應耦合電漿原子發射光譜儀(ICP-AES)
3-4-2 飼育槽設計與尺寸
3-4-3 飼育材
3-4-4 混拌之原始土壤
3-4-5 飼育材及土壤內重金屬
3-4-6 未污染幼蟲及成蟲中重金屬含量
3-5 實驗方法
3-5-1 操作管理工作流程
3-5-2 反應槽體設置
3-5-3 土壤混拌重金屬飼育材之反應槽體設置
3-5-4 重金屬採樣分析流程
3-5-5 幼蟲飼育指標說明
3-5-6 幼蟲重金屬流布採樣分析規劃
3-5-7 蟲蛹及成蟲重金屬流布採樣分析
3-6 實驗分析方法之品質管理
3-6-1 檢量線建立
3-6-2 重複實驗品質管控
3-6-3 儲備土之目標重金屬濃度測試
第四章 結果與討論
4-1 重金屬對犀角金龜幼蟲生長指標影響性
4-1-1 幼蟲對目標重金屬之降解
4-2 幼蟲每週重金屬吸收量
4-2-1 幼蟲重金屬累積吸收量
4-3 重金屬在幼蟲體內各部位之含量分布
4-4 重金屬在幼蟲體內量之質量平衡
4-5 重金屬在化蛹期及成蟲期內流布
4-6 飼育過程中重金屬整體流布探討
4-7 幼蟲進入蛹期時間及質量平衡損失率之間關係
4-7-1 重金屬流布與幼蟲成功羽化之關係
4-8 土壤混拌重金屬及飼育材對犀角金龜幼蟲生長指標影響性
第五章 結論與建議
5-1 幼蟲飼育期間
5-2 幼蟲體內重金屬流布
5-3 化蛹期及成蟲期重金屬流布
5-4 建議
第六章 工程應用性
參考文獻
附錄A
附錄B –重金屬添加對甲蟲蟲體內其它金屬的影響性
B1- 不同銅、鋅、鎳濃度飼育下，甲蟲體內其它金屬的變化
B2- 不同銅、鋅、鎳濃度飼育甲蟲脫皮及脫殼內其它金屬的變化
B3- 不同銅、鋅、鎳濃度飼育甲蟲不同部位其它金屬的變化
附錄C – 口委提問及建議

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