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標題: 柴油分解細菌對柴油乳化及分解作用之研究
Studies on Emulsification and Decomposition of Diesel by Diesel-Degrading Bacteria
作者: 吳俊德
Wu, Chun-Teh
關鍵字: diesel
出版社: 土壤環境科學系
摘要: 摘 要 石油污染的案例不勝枚舉,其發生原因不外乎經由管線、貯油槽、油輪等過程造成,其整治方法不外乎物理、化學、生物等之衍生方法,其中柴油揮發性低,使用量大,化學界面活性劑應用於油品污染的整治被認為有二次污染之虞。本研究之目的為篩選柴油分解菌及瞭解其乳化現象,以供應用於污染之整治之參考。此外本研究探討營養源添加對柴油的分解效應,並模擬柴油污染在土壤中的生物復育之可行性。 本研究所分離及篩選之柴油分解菌如Bacillus megaterium、Bacillus cereus,結果顯示之對柴油之乳化指數達52%及56 % ,而對照除菌培養液僅約6%, 顯示具有乳化之生物界面物質存在。 本研究之細菌對於不同碳源之添加對於柴油的分解效應中,以甘露糖、葡萄糖及蔗糖作為外來碳源,添加於含柴油 1 ml L-1之培養基以B. megaterium、B. cereus、Bacillus subtilis、Pseudomonas aeruginosa為分解菌。研究顯示不添加外來碳源之對照組,第12天後柴油分解率即達85 % ,添加甘露糖之處理柴油分解率約89 % , 添加葡萄糖之處理柴油分解率約為88 % , 添加蔗糖之處理柴油分解率約達88 % , 顯示添加外來碳源於柴油的分解均僅略有助益,顯示菌種對柴油之利用效果甚佳。 混合菌的柴油分解效應研究,以B. megaterium、B. cereus、 B. subtilis、Ps. aeruginosa作為單一菌株柴油分解菌,以B. megaterium、B. cereus及B. subtilis、Ps. aeruginosa作為二混合菌種,以B. megaterium、B. cereus、B. subtilis、Ps. aeruginosa作為四混合菌種進行研究,在柴油 1 ml L-1作為培養基之碳源中。結果顯示不接菌之對照組於第12天後,其分解率約 9 % , 而四株單一菌分解菌其柴油的分解率約85 % , 二混合柴油分解菌B. megaterium、B. cereus與 B. subtilis、Ps. aeruginosa其分解率約為89 % , 四混合菌其分解率達93 %。四混合菌比單一菌分解率高出約8 % , 顯示混合菌株比單一菌株有助於柴油的分解。 細菌對於不同氮源之添加對於柴油的分解效應中,以硝酸鈉與硫酸銨1 g L-1作為二種不同氮源,以1 ml L-1柴油作為碳源,經接菌 B. megaterium、B. cereus、B. subtilis、Ps. aeruginosa,結果顯示以提供硝酸鈉作為四菌株分解柴油培養基之氮源於第12天其柴油殘餘量為14 ~14.5 % ,而提供硫酸銨之處理其柴油殘餘量為13.3 ~ 14 %。故硫酸銨與硝酸鈉兩種不同氮素供給之間對於細菌的柴油分解差異並不顯著 。 柴油在土壤中之生物復育研究,本研究以B. megaterium、 B. cereus、Ps. aeruginosa、B. subtilis四混合菌為供試菌種,以二林、陳厝寮及十八股寮等三土系,進行模擬柴油污染之生物復育法試驗。由以上三種土壤研究顯示不通氣不接菌之條件下,不通氣及未接菌之對照組對柴油移出率只達3 %。陳厝寮系土壤在原有土壤微生物與四混合菌種共同作用下經土壤通氣7天後, 5 ml kg-1之柴油之移出率可達78 %。 本研究之發現在於所篩選之柴油分解菌B. megaterium、B. cereus發現對柴油具有乳化現象,並以B. megaterium、 B. cereus、Ps. aeruginosa、B. subtilis為四混合菌進行生物復育發現可有效達到整治之效果。
Abstract Petroleum pollution accidents were countless in the world. The main accidents were happened in pipeline, storage tanks and oil tankers. The relief and treatment applied were mostly physical, chemical and biological remedies. Since diesel oil has high demand for consumption and yet its volatility is pretty low. Chemical interfaces of using activator to relief and recover polluted area may afraid to give the second pollution. The objectives of this study were to select those diesel decomposing bacteria that can be used in relieving or treating hazardous petroleum pollution , as well as to find the capability of emulsify petroleum in the decomposing bacteria. Furthermore, this study has explored the impact of nutrients (C and N) on the bacterial decomposition of diesel function, and conducted simulations of bioremediation of diesel pollution in the soil . This study has screen out diesel-degrading bacteria, Bacillus megaterum, and Bacillus cereus , and found that emulsification index of diesel-degrading bacteria reached to 52% and 56%, respectively, and the bacteria removed culture solution reaches to 6 % . This indicates that exudates of besides bacteria contained is biological products that can emulsify petroleum. The assessment was made on influences of different carbon additives to the ability of decomposition of diesel. Mannitole, glucose, and sucrose were added to culture solution containing 1 ml L-1 diesel media and inoculated Bacillus megaterum, and Bacillus cereus, Pseudomonas aeruginosa. After 12th days, the control treatment without any C source showed that the decomposition of diesel was 85%, and with mannitol it was 89%, with glucose it was 88%, and with sucrose it was also about 88%. The results showed that those bacteria were effective to use the diesel in the culture. As to the effects of bacteria-mixed experiment, a single bacterium of B. megaterum, B. cereus, and Ps. aeruginosa, two bacteria mixed of B. megaterum, and B. cereus, as well as B. Subtilis and Ps. aeruginosa , and four bacteria mixed of B. megaterum, B. cereus, B. subtilis, and Ps. aeruginosa together in one were tested on diesel medium as carbon sources. In this experimentation after 12 days, control without inoculation of bacteria have exhausted 9% of diesel , while four single bacterium have exhausted 85% of diesel sources. Two bacteria mixed of B. megaterum, and B. cereus, also of , B. Subtilis and Ps. aeruginosa, both showed about 89% diesel exhaution, and finally four bacteria mix have shown carbon consumption of 93% of diesel . This result is supporting that mixed bacteria treatment decompose more diesel than single bacterium treatment. A study was made to assess influences of different nitrogen additives to the diesel degradation . 1 g L-1 sodium nitrate and ammonium sulfate were used as nitrogen sources, and , 1 ml L-1 diesel as carbon source. B. megaterum, B. cereus, and Ps. aeruginosa were used for the test. After the 12th day the residual amount of diesel was 14 — 14.5 %, while provided ammonium sulfate as nitrogen source was 13.3 — 14% .The result showed that it was not significantly different between the two different nitrogen sources. In the soil bioremediation study,the four bacteria mixed of B. megaterum, and B. cereus, B. Subtilis and Ps. aeruginosa was used in Erhlin, Chentsoliao, and Shihpakuliao series soils with 5 ml kg-1diesel as pollutant. The test of control without aeration and inoculation of bacteria showed 3 % exhaustion of diesel. Whereas soil sample from Chengkungling with four mixed bacteria have exhausted about 78 % of 5 ml kg-1 diesel after 7 days. In conclusion of the study, diesel-degrading bacteria , B. megaterum, and B. cereus , were confirmed to emulsify diesel oil and to decompose diesel. A mix of four bacteria , B. megaterum, B. cereus, B. Subtilis and Ps. aeruginosa has potential to become an effective degrading agent in bioremediation of diesel polluted soil.
Appears in Collections:土壤環境科學系



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