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年度	103	專案性質	實驗性質
專案類別	研究專案	研究主題	整治
申請機構	嘉南藥理大學	申請系所	環境工程與科學系
專案主持人	張家源	職等/職稱	教授
專案中文名稱	利用耐鹽嗜油菌與微生物富集技術強化石油碳氫化合物污染土壤之生物復育
中文關鍵字	生物復育；污染土壤；石油碳氫化合物；耐鹽嗜油菌；微生物富集整治技術
專案英文名稱	Application of salt-tolerant oil-degrading bacteria and microorganism enhancement technology on bioremediation of petroleum-hydrocarbon contaminated soil
英文關鍵字	bioremediation；soil pollution；oil hydrocarbon；salt-tolerant oil-degrading bacteria；microorganism enhancement remediation technology
執行金額
執行期間	2014/4/25 至 2014/11/24
計畫中文摘要	利用耐鹽嗜油菌與微生物富集技術強化石油碳氫化合物污染土壤之生物復 育 本研究為篩選分離柴油與甲苯降解菌，並以聚合酶連鎖反應放大菌種之 16S rDNA，進行菌種之鑑定，進而探討其生理與降解特性、乳化特性、重金屬 之吸附競爭或抑制代謝、不同營養源添加之共同代謝等，瞭解此等雙效微生 物在土壤中降解碳氫化合物與耐受重金屬銅、鋅的行為、環境影響因子等， 並應用利用實驗室集的微生物富集技術強化污染土壤之界面活性劑洗提與 生物復育運作，以提供後續實際污染環境整治的參考。 研究獲致具體成果如下： 1. 篩選分離本土甲苯降解菌 6 株，柴油降解菌 4 株，共有 10 株，甲苯降 解菌分別為 T5、T6、T7、T10、T11 與 T12；柴油降解菌分別為 D2、 D5、D6 及 D7。 2. 研究結果顯示本研究所篩選出的部分菌株具有良好的甲苯降解效能，菌 株 T6、T7 與 T11 具有較佳之甲苯降解能力。 3. 研究結果顯示本研究所篩選出的菌株具有良好的柴油降解效能，菌株 D2、D5 與 D6 具有較佳之柴油降解能力，菌株 D5 具有最佳之乳化效果。 4. 本研究篩選之甲苯分解菌均能忍受 250 ppm Zn，柴油分解菌 D2, D5 與 D7 能忍受 250 ppm Zn，高 Zn 濃度則對多數菌種產生抑制作用，其中 T7 與 D5 可忍受高濃度(500 ppm)；菌種之生長較易受銅影響，篩選出之 菌種中僅以 T7 與 D5 能耐受高濃度 Cu。本研究所分離的菌株對重金屬 之吸收有良好效能。 5. 菌種之 16S rDNA 鑑定結果發現，主要的菌種是 Achromobacter insolitus (D2)， Candida spp. (D5)，Xanthobacterpolyaromaticivorans (D6 and T12)，Ochrobactrum intermedium (D7)，Streptomyces caelestis (T1 and T5)，Gordonia amicalis (T6)，Brevibacillus brevis (T10)，Bacillus fumarioli (T11)，另外，T7 經鑑定為真菌類。 6. 本研究利用不同性質土壤，模擬甲苯或柴油污染，利用界面活性劑進行 洗提，評估界面活性劑濃度、用量、洗提時間、界面活性劑重複使用對 洗提效果之影響，結果顯示本研究選用之界面活性劑對不同性質土壤皆 有洗提效果，惟其洗提效果高低與土壤性質有關。 本研究建構實驗室微生物富集反應器系統，並於反應槽中投入本研究分 離純化的 3 種本土柴油降解菌株 (D2、D5、D6)，用以處理模擬柴油污 染土壤經界面活性劑洗提後之混合液。研究結果顯示，實驗室微生物富 集反應器系統對 COD 的處理效果達 95-97%之間，穩定且有效地處理柴 油-界面活性劑洗提液。
計畫英文摘要	Application of salt-tolerant oil-degrading bacteria and microorganism enhancement technology on bioremediation of petroleum-hydrocarbon contaminated soil This project is to screen and isolate the hydrocarbonoclastic bacteria that might be suitable for bioremediation purposes to remove toluene or diesel oil from soil in presence of zinc and copper. To achieve this goal, batch cultivation of bacteria and 16S rDNA is employed to identify the target bacteria. Several sub-study such as the characteristics of isolated bacteria, degradation performance of petroleum-hydrocarbon, biosorption of heavy metals and its inhibition of bacterial metabolism, the effects of different nutrients on co-metabolism and also a lab scale of microorganism enrichment reactor system is conducted to evaluate and enhance the treatment of surfactant washing liquid containing petroleum-hydrocarbon. 1. This project has already screened and purified 6 different kinds of toluene-degrading bacteria (T5, T6, T7, T10, T11 and T12) and 4 different kinds of diesel oil -degrading bacteria (D2, D5, D6 and D7) from different sources of oil slurry. 2. Toluene-degrading bacteria T6, T7 and T11 have the high performance on toluene degradation. 3. Diesel oil -degrading bacteria D2, D5 and D6 have the high performance on diesel degradation. 4. All toluene-degrading bacteria and diesel oil -degrading bacteria D2, D5 and D7 obtained from this study have Zn tolerance of 250 ppm, the further studies showed that T7 and D5 can tolerate high metal levels of Zn and Cu. The main mechanism of metal removal in this study is absorption. 5. Results of 16S rDNA approach showed that the dominate bacteria are Achromobacter insolitus (D2), Candida spp. (D5), Xanthobacterpolyaromaticivorans (D6 and T12), Ochrobactrum intermedium (D7), Streptomyces caelestis (T1 and T5), Gordonia amicalis (T6), Brevibacillus brevis (T10), Bacillus fumarioli (T11), strain T7 was identified as fungus by stain method. 6. The performance of surfactant on the flushing of diesel oil or toluene polluted soil was evaluated based on the parameters of surfactant concentration、volume、washing time and the reuse of surfactant. Ethylene Glycol mono-Butyl Ether is available for the flushing of CC, CF and SI soils with diesel or toluene and the performance depended on the property of soil. 7. A lab scale microorganism enrichment reactor system was conducted for the treatment of washing liquid, the results showed that the system was operated stably and a high COD removal of 97% was achieved.