生質柴油車尾氣中的多環芳香族對於大腸桿菌的基因毒性分析及代謝路徑分析

Bio diesel exhaust polycyclic aromatic for genotoxicity analysis and metabolic pathway analysis of Escherichia coli

10.6840/cycu201600947

郭仁輝

生質柴油 ； 柴油 ； 多環芳香族 ； 大腸桿菌 ； Biodiesel ； Escherichia coli ； pathway

中原大學生物環境工程研究所學位論文

2016年

碩士

黃郁慈

繁體中文

本實驗使用了大腸桿菌(E.Coli)進行與Benzo[a]pyrene標準品、Benzo[ghi]Pyrene標準品，以及中油柴油D100、生質柴油B100實車尾氣萃取的共培養試驗。 實驗中，團隊找到了這兩個樣品Benzo[a]pyrene標準品、Benzo[ghi]Pyrene標準品的半抑制率IC50分別為0.6 mg/ml、4.6 mg/ml的材料量。另外，本實驗中也取得實際添加中油柴油D100及生質柴油B100之引擎運轉後之排放尾氣PAHs萃取樣品，並對其測試微生物抑制試驗，我們發現在將吸光值為0.6A之大腸桿菌菌液1ml稀釋為原始菌液之10-5倍後，分別加入D100 1.3ml及B100 4.5ml共培養6小時後會有半抑制率IC50的產生。 後續再對這些共培養後之大腸桿菌基因層次進行進一步的生物晶片分析後，發現到Benzo(a)pyrene對於微生物之細胞膜、細胞壁、鞭毛、能量代謝等基因，有程度不一的干擾；並且在DNA損傷的基因表現上也有6倍左右的提升， Benzo(a)pyrene於多環芳香族中屬於在極少量之接觸後，會產生顯著微生物抑制、基因顯著改變的物質，於未來的實驗應該要嘗試進行更長時間的共培養條件。 另外，也應該將此次的結果與Benzo[ghi]Pyrene的基因改變量分析結果進行交互比對，尋找更深入的組合效應機制。

In this study, E. coli was used to perform co-culture experiments with benzo [a] pyrene standards, Benzo [ghi] Pyrene standards, and diesel oil D100, bio-diesel B100. In the experiment, the team found the Benzo [a] pyrene standard Benzo [ghi] pyrene standard sample semi-inhibition rate IC50 of 0.6 mg / ml, 4.6 mg / ml material volume. In addition, in this experiment, the extraction of PAHs from diesel engine D100 and bio-diesel B100 was carried out and the microbiological inhibition test was carried out. In the experiment, the concentration of 0.6% Escherichia coli solution 1ml diluted to 10-5 times the original bacteria solution, respectively, after adding D100 1.3ml and B100 4.5ml co-culture for 6 hours after the semi-inhibition rate of IC50 production. After further bioconcentration analysis of these co-cultured E. coli genomes, it was found that Benzo (a) pyrene had different degrees of interference on the cell membrane, cell wall, flagellum, energy metabolism and other genes of microorganisms; Benzo (a) pyrene is a polybrominated aromatic substance in a very small amount of contact, will produce significant microbial inhibition, gene significant changes in the material, in future experiments should be Attempts were made to co-culture conditions for a longer period of time. In addition, the results of this study should be compared with Benzo [ghi] Pyrene's gene mutation analysis results to find a more in-depth combination effect mechanism.

1. [2] MARTIN A. ELLIOTT, GERGE J. NEBEL AND FRED G. ROUNDS, The Composition of Exhaust Gases from Diesel, Gasoline and Propane Powered Motor Coaches, 2012
   連結：
2. [6] J.F. McCarthy, And B.D. Timenez, Environ. Sci. Technnol., 19 (1985) 1072.
   連結：
3. [7] Y. Cohen, Environ. Sci. Technol., 20 (1986) 38.
   連結：
4. [8] A. Bjorseth, and T. Ramdahl, Handbook of Polycyclic Aromatic Hydrocarbons, Marcel Dekker, Inc., 1985.
   連結：
5. [11] J. Josephon, “Polycyclic Aromatic Hydrocarbon”, Environ. Sci.Technol.,1984, 18, p.93A-95A
   連結：
6. [14] J. Tuominen , S. Salomss , H. Pyysalo , E. Skytta “Polynuclear Aromatic Hydrocarbons and Genotoxicity in Particulate and Vapor Phases of Ambient Air：Effect of Traffic Season,and Meteroological Conditions”, Environmental Science and Technology 22(1988) p.1228.
   連結：
7. [17] Baily, J. C., Gunary, K., Schmidl, B., Williams, M. L., 1990. Speciated hydrocarbon emissions from a sample of UK vehicles on the road over a range of Speeds, Science Total Environment 93, pp: 199-206
   連結：
8. [18] Janet Yanowitz ,Robert L. McCormick , and Michael S. Graboski, In-Use Emissions from Heavy-Duty Diesel Vehicles, Environ. Sci. Technol., 2000, 34 (5), pp 729–740
   連結：
9. [19] Hidy, G.H., Atmospheric Aerosols: Physical Characterization. Aerosols: An Industrial and Environmental Science, Academic Press, New York (1984)
   連結：
10. [20] McClellan, R.O., D.E. Bicem, R.G. Cuddiht, N.A. Gillett, R.F. Henderson, R.K. Jones, J.L. Manderly, J.A. Pickrell, S.G. Shami, R.K. Wollf, “Health Effects of Diesel Exhaust, Aerosols: Research Risk Assessment and Strategies,” Marcel Dekker, New York (1986).
    連結：
11. [22] Richter, H. and J.B. Howard, “Formation of Polycyclic Aromatic Hydrocarbons and their Growth to Soot－a Review of Chemical Reaction Pathways,” Progress in Energy and Combustion Science, Vol. 26, pp. 565-608 (2000).
    連結：
12. [23] Hays, M., N.D. Smith, J. Kinsey, Y. Dong, P. Kariher, “Polycyclic Aromatic Hydrocarbon Size Distributions in Aerosols from Appliances of Residential Wood Combustion as Determined by Direct Thermal Desorption-GC/MS,” Journal of Aerosol Science, Vol. 34, pp. 1061-1084 (2003).
    連結：
13. [26] United States Department of Agriculture (USDA) (2006), The Economic Feasibility of Ethanol Production from Sugar in the United States.
    連結：
14. [27] Kondili, E.M. and Kaldellis, J.K. (2007), “Biofuel Implementation in East Europe: Current Status and Future Prospects”, Renewable and Sustainable Energy Reviews, Vol. 11, Iss. 9, pp.2137-2151.
    連結：
15. [28] International Energy Agency (IEA) (2004), Biofuels for Transport: A International Perspective.
    連結：
16. [31] Flavin, C. (2006), “Biofuels for Transportation”, Global Potential and Implications for Sustainable Agriculture in the 21st Century, Berlin, Germany.
    連結：
17. [32] Bozbas, K. “Biodiesel as an Alternative Motor Fuel: Production and Policies in the European Union”, Renewable and Sustainable Energy Reviews [in press] DOI:10.1016/j.rser.2005.06.001.
    連結：
18. [34] Wald, M. L. (2007), “Is Ethanol for the Long Haul?”, Scientific American Magazine, January 2007 issue.
    連結：
19. [1] 盧昭暉，汽油引擎使用富氫氣體輔助燃燒之可行性探討，2007
20. [3] 劉育良，柴油引擎添加生質燃料及加裝濾煙器排放廢氣微粒之離子鹽類組成特性研究，2011
21. [4] 鄭曼婷，柴油車細懸浮微粒排放推估與減量技術之研究，98年度「環保署/國科會空污防制科研合作計畫」 成果報告，2009
22. [5] 潘一誠、林坤海，汽機車排放之臭氧前驅物及毒性空氣污染物與油品相關性之研究－期末報告，行政院環境保護署九十年度專案計畫，2001
23. [9] National Research Council, Particulate polycyclic organic matter, National Academic Science, Washington, D. C, 1972.
24. [10] W.C. Hueper, Occupational and Environmental Cancers of the Respiratory System, Springer-Verlag, New York, 1986.
25. [12] H.S. Rathore , Handbook of Chromatography- Liquid Chromatography of Polycyclic Aromatic Hydrocarbons, CRC Press, Section 1(1991) p.1-19.
26. [13] J. Longwell , P., symp. (Int.) Combust. [Proc.] 9 (1982) p.1339.
27. [15]　徐永源，汽柴油引擎車輛排放硝基多環芳香烴之調查研究，2007
28. [16] 李殷瑞、楊承憲，受多苯環化合物(PAHs)污染土壤之菌種分離及毒性代謝之研究
29. [21] Kerminen, V-.M., Mäkelä, T.E., Ojanen, C.H., Hillamo, R.E., Vilhunen, J.K.,Rantanen, L., Havers, N., Bohlen, A.V., Klockow, D., Characterization of the Particulate Phase in the Exhaust from a Diesel Car. Environmental Science and Technology, 31, 1883-1889 (1997).
30. [24] Dunlap, J., “New truck inspection programs in California aim to reduce emissions,”Diesel Progress North American Edition 64, pp. 94–96 (1998).
31. [25] 林欣儀,多環芳香烴和微粒共同暴露對細胞毒性之研究,國立陽明大學 環境衛生研究所 碩士論文,2005
32. [29] German Union for the Promotion of Oil and Protein Plants (UFOP) (2004), CO2 Mitigation through Biofuels in the Transport Sector.
33. [30] Francis, M.K. (2006), “EU-25 Sugar: The Economics of Bioethanol Production in the EU 2006”, GAIN Report Number: E36081, USDA Foreign Agricultural Service.
34. [33] Agra CEAS and Licht, F.O. (2006), “How Canada Ranks: A Comparative Study of National Biofuels Policies World-wide”, Report for the Canadian Renewable Fuels Association.