臺灣博碩士論文加值系統

現代人每天在日常生活中接觸到的一些化學物質中，少數已被證實是具有類似於生物體內荷爾蒙之作用，此物質我們俗稱為環境荷爾蒙物質(environmental hormones, EHs)。其中包括一些殺蟲劑、多氯聯苯、戴奧辛以及作為食品包裝、油漆、化妝品等之原料化合物如烷基酚類、雙酚A。由於這些物質大多無法被傳統的污水處理設備所去除，所以釵h工業及都市污水處理廠放流水中皆可發現含有這些物質的存在。因此，污水處理廠放流水也是其排放至水環境中的重要污染源之一。本研究主要目的為探討污水處理廠放流水對本土吳郭魚(Tilapia spp.)的雌激素活性。我們將魚暴露於取自污水處理廠之放流水，並以連續流(continuous flow system , CFS)之方式暴露連續達21天，其暴露濃度為稀釋50%或全放流水。雌激素活性反應評估則以魚體血清中卵黃蛋白前質(vitellogenin, VTG)的濃度來判定。另外，我們並以生殖腺體重比(gonadsomatic index, GSI)及肝臟體重比(hepatosomatic index, HSI)指數觀察魚類個體是否同時出現生理之變化。本研究結果發現：雌魚暴露在稀釋50%以及100%放流水經7天後其VTG含量可被提升；而雄魚在冬季時，暴露於稀釋50%放流水的第7天後以及暴露於春季時，100%放流水的第14天後，開始有VTG的反應。無論是雌魚或雄魚，VTG誘導的濃度皆在21天後始達到最高量，且與控制組比較後有顯著的差異。在GSI及HSI方面，暴露時期為冬、春兩季之雌魚，其GSI皆有些微增加之現象，而與控制組比較後的雄魚GSI則有顯著減小之現象；另外，暴露於各濃度放流水之雌魚或雄魚的HSI都高於對照組。由上述研究可推論污水處理廠放流水中含有具雌激素活性的化學物質，而其濃度足以誘導魚體內VTG的提升以及生理組織之變化。

A large number of substances in daily use are known to mimic natural hormones in animals. These chemicals known as environmental hormones (EHs) or endocrine disrupting chemicals (EDCs) include some pesticides, PCBs, components of food packaging materials and certain alkylphenolic substances which arise from alkylphenol polyethoxylates (APEOs) commonly used in detergents, paints and cosmetics. Sewage treatment plants (STPs) received natural and synthetic EHs from urban and industrial discharge. These compounds can not be completely removed by treatment processes of STPs and can ultimately discharged be released into the aquatic environment. Therefore, effluent from STP is an important source of EHs. The purpose of this study was to examine the effects of effluent on an indigenous freshwater fish, tilapia (Tilapia spp.). Effluent samples were collected during winter and spring and test fish were raised in aquariums and exposed to either tap water (control), half strength (50% dilution) or whole effluent for 21 days. A flow rate of 5 mL/min was applied to a continuous flow system (CFS). Estrogenic responses were evaluated based on the expression of serum vitellogenin (VTG) using western blots and quantified by an imaging analysis system. In addition, we also measured gonadosomatic index (GSI) and hepatosomatic index (HSI) to observed physiological and reproductive effects of the fish. The results showed that a short-term exposure (7 days) of the effluent at both concentrations could increase VTG levels in female during winter and spring. Male tilapia exposed to effluent at concentration of 50% dilution after 7th day has a VTG induction during winter, but during spring, male exposed to the whole effluent only after the 14th day did VTG levels elevate. The VTG levels in both male and female fish were highest at the 21th day. At the 21th day, fish were sacrificed and GSI and HSI were measured. The GSI of female fish were slightly increased, while GSI of male fish were statistically significantly smaller than control. The HSI of both female and male fish were higher than control for both experiments, but not statistically different. This study indicates that estrogenic compounds are present in STP effluents which resulting VTG induction. Concentrations of these chemicals in STP effluent were high enough to be responsible for the VTG induction and physiological alteration observed in tilapia.

目 錄
中文摘要...I
英文摘要...III
誌謝...IV
目錄...V
圖目錄...VII
表目錄...VIII
第一章 緒論...1
1-1 研究動機與目的...1
1-2 研究步驟...3
第二章 文獻回顧...6
2-1 環境荷爾蒙...6
2-1-1 概述...6
2-1-2 環境仿雌激素...14
2-1-3 污水處理廠放流水雌激素活性...16
2-1-4 烷基酚類化合物...23
2-2 生物性調查...34
2-2-1 環境荷爾蒙的檢測方法...34
2-2-2 仿雌激素與魚類之生物指標...35
第三章 研究方法...42
3-1 研究區域概述...42
3-2 實驗設計與方法...43
3-2-1 實驗設計...43
3-2-2 實驗材料與馴養條件...44
3-2-3 暴露系統...45
3-2-4 指標量測...47
3-3 西方點墨法...48
3-3-1 膠體電泳分析技術...48
3-3-2 西方轉漬...54
3-3-3 影像量化分析...59
3-4 資料分析方法...61
第四章 結果與討論...62
4-1 實驗魚存活率及健康狀況...62
4-1-1 存活率...62
4-1-2 健康狀況...63
4-2 卵黃蛋白前質誘導反應...64
4-2-1 蛋白質濃度測定...64
4-2-2 西方點墨法分析結果...76
4-2-3 影像量化分析結果...81
4-3 生殖系統指標...88
4-3-1 生殖腺體重比指數...88
4-3-2 肝臟體重比指數...91
4-4 討論...93
第五章 結論與建議...98
5-1 討論...98
5-2 結論...98
參考文獻...100


圖目錄

圖2-1 天然荷爾蒙與環境荷爾蒙作用機制...12
圖2-2 污水處理廠中壬基苯酚聚乙氧基醇類化合物(NPnEO)流佈...22
圖2-3 壬基苯酚聚乙氧基醇類遭水環境中微生物分解之降解途徑...27
圖2-4 魚類生殖內分泌系統...37
圖3-1 污水處理廠區域圖...43
圖3-2 連續流暴露系統...46
圖3-3 膠體電泳法來及西方轉漬流程圖...58
圖4-1 各暴露時間及暴露濃度實驗魚存活率...62
圖4-2 冬季組各暴露時間蛋白質濃度檢量線...66
圖4-3 春季組各暴露時間蛋白質濃度檢量線...71
圖4-4 冬季組吳郭魚血清VTG影像圖(暴露前)...77
圖4-5 冬季組吳郭魚血清VTG影像圖(暴露7天)...77
圖4-6 冬季組吳郭魚血清VTG影像圖(暴露14天)...78
圖4-7 冬季組吳郭魚血清VTG影像圖(暴露21天)...78
圖4-8 春季組吳郭魚血清VTG影像圖(暴露前)...79
圖4-9 春季組吳郭魚血清VTG影像圖(暴露7天)...79
圖4-10 春季組吳郭魚血清VTG影像圖(暴露14天)...80
圖4-11 春季組吳郭魚血清VTG影像圖(暴露21天)...80
圖4-12 冬季吳郭魚血清卵黃蛋白前質誘導之劑量與反應關係...86
圖4-13 春季吳郭魚血清卵黃蛋白前質誘導之劑量與反應關係...87
圖4-14 劑量反應關係實驗之生殖腺體重比指數...90
圖4-15 劑量反應關係實驗之肝臟體重比指數.....92


表目錄

表2-1 我國相關法規管理疑似環境荷爾蒙化學物質情形一覽表...8
表2-2 天然及人造雌激素化合物代謝比較表...16
表2-3 各國污水處理廠放流水中天然及合成雌激素的濃度範圍及平均值...18
表2-4 各國雌激素暴露的野外及半野外試驗中魚類VTG的含量...19
表2-5 污水處理廠放流水及承受水體中NP(μg/L)含量...20
表2-6 烷基酚類化合物一覽表...24
表2-7 烷基酚類化合物在各國的使用量...25
表2-8 各國河川中壬基苯酚及其衍生物之濃度比較...30
表2-9 台灣河川壬基苯酚化合物檢出濃度值...32
表2-10 壬基苯酚化合物與天然雌二醇(17β-Estradiol)之相對雌激素效應...34
表3-1 劑量與反應關係實驗設計一覽表...44
表3-2 配置0.75mm電泳膠片所需之配方...51
表4-1 冬季不同放流水濃度及暴露時間對實驗魚健康狀況之影響...63
表4-2 春季不同放流水濃度及暴露時間對實驗魚健康狀況之影響...64
表4-3 冬季組吳郭魚血清蛋白質濃度表(實驗前)...67
表4-4 冬季組吳郭魚血清蛋白質濃度表(第7天)...68
表4-5 冬季組吳郭魚血清蛋白質濃度表(第14天)...69
表4-6 冬季組吳郭魚血清蛋白質濃度表(第21天)...70
表4-7 夏季組吳郭魚血清蛋白質濃度表(實驗前)...72
表4-8 夏季組吳郭魚血清蛋白質濃度表(第7天)...73
表4-9 夏季組吳郭魚血清蛋白質濃度表(第14天)...74
表4-10 夏季組吳郭魚血清蛋白質濃度表(第21天)...75
表4-11 冬季吳郭魚血清卵黃蛋白前質光學量化強度...85
表4-12 春季吳郭魚血清卵黃蛋白前質光學量化強度...85
表4-13 實驗終點生殖腺體重比與肝臟體重比指數...89
表4-14 露條件與VTG誘導的關係...96

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