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研究生:林惠元
研究生(外文):Huei-Yuan Lin
論文名稱:安姆氏法評估環境荷爾蒙類農藥之致變異性
論文名稱(外文):The mutagenic effect of environmental hormone pesticides evaluated by Ames test assay
指導教授:王順成
指導教授(外文):Shun-Cheng Wang
學位類別:碩士
校院名稱:朝陽科技大學
系所名稱:環境工程與管理系碩士班
學門:工程學門
學類:環境工程學類
論文種類:學術論文
論文出版年:2011
畢業學年度:99
語文別:中文
論文頁數:120
中文關鍵詞:致變異性環境荷爾蒙除草劑殺菌劑Ames test殺蟲劑
外文關鍵詞:Ames testinsecticidemutationenvironmental hormonesherbicidefungicide
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近年來,許多含有疑似環境荷爾蒙成份之農藥相繼被檢測出,而此些農藥可能兼具致變異性,當人體長期暴露於低劑量下,易於體內累積並造成慢性疾病、生殖系統疾病或致癌等影響,因此逐漸引起國際間的關注。本研究目的是利用Ames test之非代謝型遺傳致變異方法輔以化學分析,快速檢測本試驗選定之台灣常用十三種被列為疑似環境荷爾蒙農藥的原體及商品,包括:歐殺松(Acephate)、達馬松(Methamidophos)、益達胺(Imidacloprid)、阿巴汀(Abamectin)、納乃得(Methomyl)、加保利(Carbaryl)、賽滅寧(Cypermethrin)、依普同(Iprodione)、三泰芬(Triadimefon)、四氯異苯腈(Chlorothalonil)、嘉磷塞(Glyphosate)、莫多草(Metolachlor)及草滅淨(Simazine)是否兼具致變異性,並另外採取農藥工廠廢水,包括:原水、生物舖氣水、曝氣水及放流水,分析農藥之複合致變異性,此等資料,可提供進一步動物試驗之參考或根據。結果顯示,部分疑似環境荷爾蒙農藥且兼具致變異性之藥劑,原體部分為:加保利、三泰芬;商品部分為:加保利、賽滅寧、益達胺、依普同、三泰芬、四氯異苯腈及嘉磷塞,農藥工廠採樣廢水,包含:原水、曝氣水及放流水,檢測結果具致變異性,應注意廢水排放標準,以免造成環境生態衝擊。
In recent years, many pesticides have been detected containing suspected environmental hormones. These pesticides may also contain variability. This kind of pesticides could easily accumulate in human body and then further chronic diseases, reproductive system diseases, and cancer under low dosage and long exposure period. As above mentioned, it attracts much attention about this issue worldwide. The aim of this study was to use non-metabolic mutagenic Ames test and chemical analysis to rapidly detect thirteen suspected environmental hormone pesticides conventionally used in Taiwan including Acephate, Methamidophos, Imidacloprid, Abamectin, Methomyl, Carbaryl, Cypermethrin, Iprodione, Triadimefon, Chlorothalonil, Glyphosate, Metolachlor and Simazine. In addition, we also collected waste water discharged from pesticide factories including raw water, bio-aerated water, aerated water and drained water to analyze the composite mutagenesis of pesticides. According to the results above mentioned, it could provide references or bases for further animal experiments. The results revealed that partial suspected environmental pesticides with mutagenesis could be separated into the technical grade material parts, i.e., Carbaryl and Triadimefon, commercial parts, i.e., Carbaryl, Cypermethrin, Imidacloprid, Iprodione, Triadimefon, Chlorothalonil and Glyphosate, and waste water discharged from pesticide factories including raw water, aerated water and drained water. Owing to environmental hormone pesticides above mentioned with mutagenesis, we should notice rules about waste water discharges to prevent environmental and ecological damage.
目錄
摘要.....................................................................................................................Ⅰ
英文摘要.............................................................................................................Ⅱ
誌謝.....................................................................................................................Ⅳ
目錄…………………………………………………………………………….Ⅴ
表目錄……………………………………………………………………….…Ⅶ
圖目錄……………………………………………………………………….. XIV
第一章 前言…………………………………………………………………….1
第二章 文獻回顧……………………………………………………………….4
一、農藥之應用與安全性...........................................................................4
二、農藥於環境中之移動與降解情形…………………………………...7
三、環境荷爾蒙對人體之作用機制…………………………………….11
四、疑似環境荷爾蒙農藥之毒理性質………………………………….15
五、農藥之化學添加物及不純物探討………………………………….29
六、安姆氏試驗法之應用.........................................................................31
第三章 研究目的...............................................................................................34
第四章 材料與方法…………………………………………………………...36
一、材料………………………………………………………………….36
(一)、供試菌種…………………………………………………….36
(二)、供試藥劑…………………………………………………….38
(三)、試驗儀器…………………………………………………….40
(四)、供試材料…………………………………………………….41
二、方法………………………………………………………………….41
(一)、Ames test原理........................................................................41
(二)、培養基及溶液配製………………………………………….42
(三)、鼠傷寒沙門氏菌株培養方法……………………………….44
(四)、細菌數計算法……………………………………………….44
(五)、供試藥劑之處理…………………………………………….45
(六)、試驗步驟…………………………………………………….46
(七)、結果判定與品保…………………………………………….47
(八)、台灣A地區農藥工廠廢水採樣……………………………50
第五章 結果與討論………………………………………………………….. 51
一、疑似環境荷爾蒙殺蟲劑之致變異性……………………………….51
二、疑似環境荷爾蒙殺菌劑之致變異性……………………………….77
三、疑似環境荷爾蒙除草劑之致變異性……………………………….89
四、台灣A地區農藥工廠採樣廢水之複合致變異性及化學分析……97
第六章 結論………………………………………………………………….104
第七章 參考文獻…………………………………………………………….105
表目錄
表1. 毒性化學物質分類管理架構....................................................................6
表2. 美國環保署2009年公佈第一階段疑似環境荷爾蒙最終篩選清單之部
份藥劑(為本試驗之選用藥劑)..................................................................7
表3. 農藥於環境中之殘效性.............................................................................9
表4. 有機化合物之吸附係數及移動性等級....................................................10
表5. 地下水污染指數........................................................................................11
表6. 政府限制登記使用之殺蟲劑....................................................................28
表7. 政府限制登記使用之殺菌劑...................................................................29
表8. 政府限制登記使用之除草劑...................................................................29
表9. 幾種短期致變異性試驗方法對已知致變劑及致癌劑之預測率……...33
表10. 常用之沙門氏菌菌株基因型.................................................................36
表11. 沙門氏菌株自然回復突變數控制值.....................................................49
表12. 疑似環境荷爾蒙之加保利(Carbaryl)殺蟲劑(原體)對沙門氏菌TA98之回復突變菌落數................................................................................54
表13. 疑似環境荷爾蒙之加保利(Carbaryl)殺蟲劑(原體)對沙門氏菌TA100之回復突變菌落數................................................................................54
表14. 疑似環境荷爾蒙之加保利(Carbaryl)殺蟲劑(商品)對沙門氏菌TA98之回復突變菌落數................................................................................55
表15. 疑似環境荷爾蒙之加保利(Carbaryl)殺蟲劑(商品)對沙門氏菌TA100之回復突變菌落數................................................................................55
表16. 疑似環境荷爾蒙之益達胺(Imidacloprid)殺蟲劑(原體)對沙門氏菌TA98之回復突變菌落數......................................................................58
表17. 疑似環境荷爾蒙之益達胺(Imidacloprid)殺蟲劑(原體)對沙門氏菌TA100之回復突變菌落數………………………………………........58
表18. 疑似環境荷爾蒙之益達胺(Imidacloprid)殺蟲劑(商品)對沙門氏菌TA98之回復突變菌落數……………………………………………..59
表19. 疑似環境荷爾蒙益達胺(Imidacloprid)殺蟲劑(商品)對沙門氏菌TA100之回復突變菌落數…..………………………………………..59
表20. 疑似環境荷爾蒙之納乃得(Methomyl)殺蟲劑(原體)對沙門氏菌TA98之回復突變菌落數……………………………………………………63
表21. 疑似環境荷爾蒙之納乃得(Methomyl)殺蟲劑(原體)對沙門氏菌TA100之回復突變菌落數……………………………………………………63
表22. 疑似環境荷爾蒙之納乃得(Methomyl)殺蟲劑(商品)對沙門氏菌TA98之回復突變菌落數…………………………………………………....64
表23. 疑似環境荷爾蒙之納乃得(Methomyl)殺蟲劑(商品)對沙門氏菌TA100之回復突變菌落數…………………………………………....64

表24. 疑似環境荷爾蒙之歐殺松(Acephate)殺蟲劑(原體)對沙門氏菌TA98之回復突變菌落數…………………………………………………....65
表25. 疑似環境荷爾蒙之歐殺松(Acephate)殺蟲劑(原體)對沙門氏菌TA100之回復突變菌落數…...……………………………………………….65
表26. 疑似環境荷爾蒙之歐殺松(Acephate)殺蟲劑(商品)對沙門氏菌TA98之回復突變菌落數………………...………………………………….66
表27. 疑似環境荷爾蒙之歐殺松(Acephate)殺蟲劑(商品)對沙門氏菌TA100之回復突變菌落數…...……………………………………………….66
表28. 疑似環境荷爾蒙之達馬松(Methamidophos)殺蟲劑(原體)對沙門氏菌TA98之回復突變菌落數………….………………………………….67
表29. 疑似環境荷爾蒙之達馬松(Methamidophos)殺蟲劑(原體)對沙門氏菌TA100之回復突變菌落數………...………………………………….67
表30. 疑似環境荷爾蒙之達馬松(Methamidophos)殺蟲劑(商品)對沙門氏菌TA98之回復突變菌落數……….…………………………………….68
表31. 疑似環境荷爾蒙之達馬松(Methamidophos)殺蟲劑(商品)對沙門氏菌TA100之回復突變菌落數………...………………………………….68
表32. 疑似環境荷爾蒙之賽滅寧(Cypermethrin)殺蟲劑(原體)對沙門氏菌TA98之回復突變菌落數…….……………………………………….71

表33. 疑似環境荷爾蒙之賽滅寧(Cypermethrin)殺蟲劑(原體)對沙門氏菌TA100之回復突變菌落數……...…………………………………….71
表34. 疑似環境荷爾蒙之賽滅寧(Cypermethrin)殺蟲劑(商品)對沙門氏菌TA98之回復突變菌落數…….……………………………………….72
表35. 疑似環境荷爾蒙之賽滅寧(Cypermethrin)殺蟲劑(商品)對沙門氏菌TA100之回復突變菌落數…...……………………………………….72
表36. 疑似環境荷爾蒙之阿巴汀(Abamectin)殺蟲劑(原體)對沙門氏菌TA98之回復突變菌落數…………………...……………………………….75
表37. 疑似環境荷爾蒙之阿巴汀(Abamectin)殺蟲劑(原體)對沙門氏菌TA100之回復突變菌落數…...……………………………………….75
表38. 疑似環境荷爾蒙之阿巴汀(Abamectin)殺蟲劑(商品)對沙門氏菌TA98之回復突變菌落數……………………………………………………76
表39. 疑似環境荷爾蒙之阿巴汀(Abamectin)殺蟲劑(商品)對沙門氏菌TA100之回復突變菌落數…………………………………………….76
表40. 疑似環境荷爾蒙之依普同(Iprodione)殺菌劑(原體)對沙門氏菌TA98之回復突變菌落數……………………………………………………79
表41. 疑似環境荷爾蒙之依普同(Iprodione)殺菌劑(原體)對沙門氏菌TA100之回復突變菌落數……………………………………………………79

表42. 疑似環境荷爾蒙之依普同(Iprodione)殺菌劑(商品)對沙門氏菌TA98之回復突變菌落數……...…………………………………………….80
表43. 疑似環境荷爾蒙之依普同(Iprodione)殺菌劑(商品)對沙門氏菌TA100之回復突變菌落數…………...……………………………………….80
表44. 疑似環境荷爾蒙之四氯異苯腈(Chlorothalonil)殺菌劑(原體)對沙門氏菌TA98之回復突變菌落數………...………………………………..83
表45. 疑似環境荷爾蒙之四氯異苯腈(Chlorothalonil)殺菌劑(原體)對沙門氏菌TA100之回復突變菌落數……...…………………………………83
表46. 疑似環境荷爾蒙之四氯異苯腈(Chlorothalonil)殺菌劑(商品)對沙門氏菌TA98之回復突變菌落數……………..…………………………...84
表47. 疑似環境荷爾蒙之四氯異苯腈(Chlorothalonil)殺菌劑(商品)對沙門氏菌TA100之回復突變菌落數………………...………………………84
表48. 疑似環境荷爾蒙之三泰芬(Triadimefon)殺菌劑(原體)對沙門氏菌TA98之回復突變菌落數…..…………………………………………87
表49. 疑似環境荷爾蒙之三泰芬(Triadimefon)殺菌劑(原體)對沙門氏菌TA100之回復突變菌落數……………………………………………87
表50. 疑似環境荷爾蒙之三泰芬(Triadimefon)殺菌劑(商品)對沙門氏菌TA98之回復突變菌落數……………………………………………..88

表51. 疑似環境荷爾蒙之三泰芬(Triadimefon)殺菌劑(商品)對沙門氏菌TA100之回復突變菌落數……………………………………………88
表52. 疑似環境荷爾蒙之嘉磷塞(Glyphosate)除草劑(原體)對沙門氏菌TA98之回復突變菌落數……………………………………………………91
表53. 疑似環境荷爾蒙之嘉磷塞(Glyphosate)除草劑(原體)對沙門氏菌TA100之回復突變菌落數……………………………………………91
表54. 疑似環境荷爾蒙之嘉磷塞(Glyphosate)除草劑(商品)對沙門氏菌TA98之回復突變菌落數……………………………………………………92
表55. 疑似環境荷爾蒙之嘉磷塞(Glyphosate)除草劑(商品)對沙門氏菌TA100之回復突變菌落數……………………………………………92
表56. 疑似環境荷爾蒙之草滅淨(Simazine)除草劑(原體)對沙門氏菌TA98之回復突變菌落數……………………………………………………95
表57. 疑似環境荷爾蒙之草滅淨(Simazine)除草劑(原體)對沙門氏菌TA100之回復突變菌落數……………………………………………………95
表58. 疑似環境荷爾蒙之莫多草(Metolachlor)除草劑(原體)對沙門氏菌TA98之回復突變菌落數..……………………………………………96
表59. 疑似環境荷爾蒙之莫多草(Metolachlor)除草劑(原體)對沙門氏菌TA100之回復突變菌落數……………………………………………96

表60. 台灣A地區農藥工廠採樣廢水對沙門氏菌TA98之回復突變菌落數
………………………………………………………………………..100
表61. 台灣A地區農藥工廠採樣廢水對沙門氏菌TA100之回復突變菌落數
…………………………………………………………………….….100
表62. 台灣A地區農藥工廠採樣廢水之化學分析…………………….…100
表63. 疑似環境荷爾蒙農藥之特性與致變異情形…………………….….101

圖目錄
圖1. 農藥在環境中之移動情形........................................................................8
圖2. 典型正常生物激素(荷爾蒙)之結構與作用機制....................................14
圖3. 環境荷爾蒙干擾雌性激素之作用機制...................................................14
圖4. 環境荷爾蒙干擾雄性激素之作用機制...................................................15
圖5. 化學致突變物之毒性作用........................................................................16
圖6. 化學因子致癌之多步驟............................................................................16
圖7. 研究架構...................................................................................................35
圖8. 鼠傷寒沙門氏菌鹽基置換之作用機制...................................................37
圖9. 鼠傷寒沙門氏菌框構位移之作用機制...................................................37
圖10. 藥劑平板混合試驗流程..........................................................................47
圖11. 圖14. TA98及TA100空白及正反應對照組織菌落回復突變數
(未加S9)..................................................................................................50
圖12. 台灣A地區農藥工廠廢水處理流程......................................................51
圖13. 本研究疑似環境荷爾蒙農藥殺蟲劑之化學結構式...................... .....102
圖14. 本研究疑似環境荷爾蒙農藥殺菌劑之化學結構式............................103
圖15. 本研究疑似環境荷爾蒙農藥除草劑之化學結構式............................103
方麗萍,「全球農藥及台灣除草劑市場之概況多樣性」,農藥一路發電子報,第20345期,第1頁(2010)。
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