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標題: 柴油分解細菌之分離鑑定與特性分析
Isolation, Identification and Characterization of Diesel-Degrading Bacteria
作者: 許伊含
Hsu, Yi-Han
關鍵字: duesel contaminated
出版社: 土壤環境科學系
摘要: 石化工業的產物被認為是最常污染土壤及地下水的有機污物, 而柴油是石油分餾產物之一,因而對柴油污染的整治是刻不容緩的事情。過去十幾年,生物復育技術蓬勃發展,常用來清除被污染土壤中的危險性化合物。因此,本研究的目的為由受柴油污染土壤中,分離出具柴油降解能力的細菌,並觀察細菌在水相環境中對柴油的乳化能力及降解能力。實驗最初由被柴油污染的土壤中,以柴油為唯一碳源分離出數十株菌株。將經由初步篩選後所得的二十株菌株,以16S rDNA序列鑑定菌種,並觀察菌對柴油的乳化能力及降解能力。初步篩選而得的二十株菌株,皆能在三天以內,於以柴油為唯一碳源的培養基上形成菌落,且能在含柴油環境中存活。細菌基因鑑定結果顯示,菌株分別為Pseudomonas aeruginosa,Pseudomonas stutzeri,Acidovorax temperans,Acinetobacter radioresistens,Sphingomonas paucimobilis及Acinetobacter lwoffii六個菌種。在菌的乳化能力試驗中,未加菌之空白處理的乳化指數為1%:除了菌株c、e及G以外的十六株菌株,對柴油皆有乳化能力,其中以菌株1-2(Pseudomonas aeruginosa)在第一天的乳化指數最高,達到30%;菌株c、e及G三天的乳化能力皆與空白處理皆無差異,因此這三株菌株對柴油沒有乳化能力。在菌對柴油的降解實驗中,最初的柴油量皆為4912 mg L-1。未接菌的空白處理之柴油逸失量約為4%,每株菌與空白處理比較,皆對柴油有降解能力,其中以菌株5 (Pseudomonas stutzeri)的降解能力最佳。菌株5在經過十五天的培養後,降解了40%的柴油;菌株3-1(Acidovorax temperans)在經過十五天的培養後,對柴油的降解率為15%;菌株V(Acinetobacter lwoffii)在培養十四天後,降解了19%的柴油;菌株W(Acinetobacter radioresistens)在經過十八天的培養後,降解了12%的柴油;而菌株G(Sphingomonas paucimobilis)在培養十九天後,約利用了12%的柴油。因此,本研究中所分離出的菌株3-1、V、W、5及G對柴油皆有降解能力,其中菌株5的降解能力是這五株菌株中最高的。
The products of petroleum industry are considered as the most frequent organic pollutants of soil and ground water. Because diesel fuel is one of the fractional distillation products, it's of great urgency to straighten diesel-contaminated site. The past decade, bioremediation techniques have been developed and improved to clean up soils polluted with hazardous chemicals. The aim of this work is to isolate diesel- degrading bacteria from diesel-contaminated soil, and to investigate the diesel-emulsification and diesel-degradation ability of these strains in the aquatic environment. Originally, isolated several bacteria using the selective medium which have diesel as the unique C source. Followly, identified these initial-screening strains by 16S rDNA sequencing, and investigated their diesel-emulsification and diesel-degrading abilities. The initial-screening strains formed colony on the medium in 3 days, and could survive under the high-concertration-diesel condition. The gene identification results showed that these strains belong to Pseudomonas aeruginosa, Pseudomonas stutzeri, Acidovorax temperans, Acinetobacter radioresistens, Sphingomonas paucimobilis and Acinetobacter lwoffii. In the diesel-emulsification experiment, the emulsion index of uncultured control was 1%. Among 19 strains, strain c, e, G had no diesel emulsification ability because of no variation with control in three days. The others could emulsify diesel and strain 1-2 (Pseudomonas aeruginosa) had highest emulsion index in the first day up to 30%. Compared with uncultured control, every strain had ability to utilize diesel in the diesel-degrading experiment, and strain 5 (Pseudomonas stutzeri) had best diesel- degrading ability. Strain 5 degraded 40% diesel after 15 days of incubated. Strain 3-1 (Acidovorax temperans) degraded 15% diesel after 15 days of incubated. Strain V (Acinetobacter lwoffii) degraded 14% diesel after 19 days of incubated. Strain W (Acinetobacter radioresistens) degraded 18% diesel after 12 days of incubated. And strain G (Sphingomonas paucimobilis) degraded 12% diesel after 19 days of incubated. Hence, the isolated strains 3-1, V, W, 5 and G of this work can utilize diesel, and strain 5 had best ability to degrade diesel.
Appears in Collections:土壤環境科學系



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