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年度	112	專案性質	實驗性質
專案類別	模場試驗	研究主題	整治
申請機構	國立中央大學	申請系所	生命科學系
專案主持人	陳師慶	職等/職稱	特聘教授
專案中文名稱	開發高效本土脫氯菌劑及優化現地生物加強傳輸工法：現地模場試驗
中文關鍵字	脫鹵球菌、生物加強、生物整治、三氯乙烯、完全脫氯
專案英文名稱	Develop high-efficiency dechlorination consortia and optimize the bioaugmentation strategy of bioremediation in chlorinated contamination: pilot study
英文關鍵字	Dehalococcoides, bioaugmentation, bioremediation, trichloroethene,completed dechlorination
執行金額	2,250,000元
執行期間	2023/9/1 至 2024/8/31
計畫中文摘要	含氯有機物如三氯乙烯為難降解之污染物，常污染我國地下水並且整治難度高，污染場址的解列率僅不到兩成。目前現地整治遇到最大的瓶頸是業者多使用化學藥劑進行生物整治，整治最終導致毒性更強並能致癌的氯乙烯(Vinyl chloride)的累積，這個因素常常讓一個場地的整治失敗。本團隊透過過去與產業端合作，收集全台含氯有機物污染之地下水，進一步培養台灣本土脫氯菌群，使用本團隊獨有的脫鹵球菌(Dehalococcoides)培養平台，我們成功地找到數個菌群可以快速且不易產生二次污染如氯乙烯的菌群，用以開發創新本土菌劑生物強化(Bioaugmentation)整治工法。本研究第一年度，透過本實驗室從現地模場建立的厭氧脫氯菌群資料庫中，去開發應用於現地的脫氯菌劑，我們將會在這個計畫中去使用發酵槽開發量產製程，使得菌量達可以達應用於現地整治之標準，同時確立脫氯菌劑的降解效能。於期中報告中，本團隊開發一不易累積氯乙烯之菌群，該菌群不含致病菌，具有豐富產氫菌及眾多發酵者，適合應用於地下水環境，由此菌群所開發之脫氯菌劑製程可以以40天培養週期穩定量產，每一批次可以生產100公升，脫鹵球菌菌數可達1011 cells/L，包含脫氯功能性基因tceA以及vcrA。本計畫以雙環塞灌注工法進行脫氯菌劑現地生物強化，加入菌劑後 3 個月現地地下水之脫鹵球菌菌數、tceA 以及 vcrA 皆有所增長，污染物降解率可達80%。本計畫第二年度持續觀察污染物降解情形，針對脫鹵球菌菌數較低的灌注井進行重力灌注補強。重力灌注後，污染物成功降解並低於監測標準，在脫鹵球菌充足情況下，使得污染物濃度不容易在回升，顯示菌種的活性可維持長達一年。持續觀察地下水菌相變化情形，發現灌注後菌劑菌種會暫時成為現地的優勢菌群，但在污染物降解完畢後，現地地水微生物菌群將逐漸回復至原始生態，以上成果證明使用本土脫鹵球菌菌劑結合 DPI 灌注工法之可行性及環境友善性。最終，使用本工法進行整治，已成功降解場址目標污染物1,1-二氯乙烯，降解率高達96 %，與傳統生物刺激工法進行效益比較，使用生物菌劑進行整治效益將提升整體效益 194%，整治大幅縮短整治時程且能達到完全脫氯，因此，我們為台灣提供一項嶄新脫氯菌劑生物強化工法，以提升我國多氯乙烯污染整治之能力。
計畫英文摘要	Organic halide compounds such as trichloroethyene are refractory pollutants, which often pollute the groundwater in our country and are difficult to remediate. The decommissioning rate of polluted sites is less than 20%. At present, the biggest bottleneck encountered in remediation is that operators often use chemical agents for biological remediation. Remediation eventually leads to the accumulation of vinyl chloride, which is more toxic and carcinogenic. This factor often makes the remediation of a site fail. Through cooperation with the industry in the past, our team has collected groundwater contaminated by chlorinated organic compounds throughout Taiwan, and further cultivated Taiwan's native dechlorination bacteria. Using our unique Dehalococcoides culture platform, we have successfully found several consortia. These consortia can quickly and less produce secondary pollution such as vinyl chloride, to develop innovative novel bioaugmentation strategies. In the first year of this project, through the laboratory of the anaerobic dechlorination bacteria database established by the on-site model field, we will develop the dechlorination consortia used in the field. We will use the fermentation tank in this project to develop. The production process enables the consortia to reach the standard that can be applied to on-site remediation, and at the same time establishes the degradation efficiency of the dechlorination bacteria agent. In the second year of this project, we continued to monitor the degradation of pollutants and reinforced injection wells with low Dehalococcoides counts by performing gravity injections. After these gravity injections, pollutants were successfully degraded to levels below the monitoring standards. Although a temporary rebound of pollutants occurred during the process, they were effectively degraded again when Dehalococcoides counts were sufficient, indicating that the activity of the bacterial strains could be sustained for up to a year. Ongoing monitoring of groundwater microbial communities revealed that the injected bacterial strains altered the local microbiota, but after the pollutants were completely degraded, the local microbial community gradually returned to its original state. This demonstrates the feasibility and environmental friendliness of using native Dehalococcoides combined with the DPI injection method. Ultimately, this method successfully degraded the target pollutant, 1,1-dichloroethylene, with a degradation rate of 96%, leading to the successful delisting of the Wanbao site. This innovative dechlorination bioaugmentation method provides a new approach to enhance the remediation of chlorethenes pollution in Taiwan.