本研究主要目的為從重油污染土壤中，篩選出對重油具降解能力的細菌，並對此細菌進行菌種鑑定及特性探討。研究中共採集八個來自中油儲油槽及煉油廠鄰近受到重油污染的土壤樣本，採集之土壤經由馴養後，測試土壤中細菌對重油的降解力。研究結果顯示土壤中混合菌(#6)可於十天後將2,000 ppm重油降解至572 ppm。從具有重油降解能力的混合菌種培養液中純化分離，挑選出三株在DGGE膠片上不同位置的菌種，進行16S rDNA定序及微生物生化測試鑑定菌種。鑑定結果顯示此三株菌分別為：Pseudomonas aeruginosa (NSYSU-1-1), Acinetobacter sp.(NSYSU-4-1), 及 Pseudomonas sp.(NSYSU-7-1)。以此三株菌種、三株菌混合(mix)與混合菌種#6進行最適降解條件測試，在大部份的處理條件下，以污染土樣放大培養的混合菌#6對重油的降解效果較單一菌株佳。溫度對微生物降解重油的影響較不明顯。氧氣含量培養在高轉速(160 rpm) 較低轉速(20 rpm)條件下有較佳的降解效率，可提早10天降解重油至1,000 ppm之土壤管制標準之下。菌種NSYSU-1-1及NSYSU-7-1雖然都屬於Pseudomonas菌屬，但在不同的酸鹼度及氧氣含量的環境下降解效果皆不相同。本實驗菌種在氮源增加的條件下可以明顯的加強重油降解能力，磷源增加對重油降解作用影響較不顯著。在最適的處理條件下，篩選出的菌種皆可降解重油至1000 ppm之土壤管制標準之下。本研究所測試的最佳降解菌種及處理條件等技術，未來可作為污染整治實場處理上之參考。

The purpose of this study is to isolate and identify the crude oil-degrading bacteria from oil polluted soil. Their physiological characteristics and oil-degrading capability were also studied. Eight polluted soil samples were taken from the Kaohsiung Refingery Factory of the Chinese Petroleum Corporation (CPC). The microbiota of the Kaohsiung refinery soil sample P37-2 (#6) could degrade crude oil from 2000 ppm to 572 ppm in 10 days. Bacteria in polluted soil samples were selected and isolated by minimal medium with 2000 ppm crude oil as the sole carbon source. Biochemical test, PCR-DGGE, and 16S DNA sequencing were used to identify and characterize the bacteria isolates. Three strains were identified as Pseudomonas aeruginosa (NSYSU-1-1), Acinetobacter sp. (NSYSU-4-1), and Pseudomonas sp. (NSYSU-7-1). These three strains and microbiota #6 were tested for their capability of degrading the total petroleum hydrocarbons (TPH). We found that microbiota #6 performed better than the other three bacterial strains in degrading the crude oil. In this study, we also found temperature was not the major factor of influcing the biodegradation; however, high oxygen concentration and providing nitrogen soure couled improve the biodegradation rate. Although both NSYSU-1-1 and NSYSU-7-1 are Pseudomonas strains, they performed different on degrading the oil. All strains tested could degrade the crude oil to a concentration below 1000 ppm to meet the government emission standard. The bacterial strains and techniques developed in this study provide a choice for future bioremediation of crude oil pollution.

中文摘要.................................................Ⅱ
Abstract.................................................Ⅲ
目錄.....................................................Ⅳ
圖目錄...................................................Ⅳ
表目錄...................................................Ⅸ
前言......................................................1
材料與方法...............................................17
實驗結果與討論...........................................31
總結.....................................................59
參考文獻.................................................61
圖表.....................................................67
附錄....................................................101

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