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.