長期受總石油碳氫化合物(TPH)汙染之土壤，在生物復育過程中往往面臨殘餘濃度過高、復育時間過長等問題，原因係降解過程中發生遲滯現象，且無法藉自然的程序進行第二階段降解的反應。本研究目的係針對長期受總石油碳氫化合物汙染之土壤，建立兩階段生物整治策略，縮短生物降解遲滯時間，並快速推進第二階段降解，達到殘餘濃度去除的目的。
為了達到此目的，本研究係以初始濃度14,032 mg/kg dry soil之長期汙染土壤，結合生物菌劑及助劑於生物降解策略做降解最佳化探討，即於此TPH汙染土壤中添加108 CFU/g dry soil的五株優菌株，配合濃度100 mg/kg soil之生物界面活性劑rhamnolipid (A8R100)，可以在35天達到第一階段降解，去除率高達70%。至於各組降解遲滯的問題則與汙染物三成份中極性物質的累積有關，在累積過程中真菌類多樣性的增加，有助於極性物質的去除，最後細菌類與真菌類共生的環境能使汙染物產生第二階段的降解反應，因此汙染物能持續降解。
由實驗室多重土堆生物整治結果，設計一套兩階段整治策略程序，以A8R100策略促進第一階段汙染物的降解，當TPH遲滯現象發生時則輔以真菌類多樣性之策略來增加極性物質的降解，可縮短遲滯期的時間。結果發現，以A8R100應用在兩階段生物復育程序，確實能將初始濃度7,379 mg/kg dry soil之土壤，在21天內達到第一階降解(去除率為65%)；再於第二階段添加優植油解真菌能將去除率提昇至87%，單以酵母菌的優植方式也可以達到89%的去除效果，而以添加廚餘堆肥的策略，更能將汙染物提升到89%以上。以此兩階段的組合於三個月內即能達到汙染物低於法規標準1,000 mg/kg dry soil，藉由優植真菌及廚餘堆肥的策略才能減少遲滯現象的發生，並快速達到汙染物降解的目的。

Soils which are long term contaminated with petroleum hydrocarbons usually encountered with problems of high residual concentrations and usually require a relatively long remediation time. The reason can be the lag phase observed during the remediation course, as well as the difficulty of natural TPH degradation during the second-stage remediation process.
The objective of this study was to establish a two-stage bioremediation strategy to remediate an aged hydrocarbon-contaminated soil. The purpose of the strategy was to reduce the time taken for the 1st-stage degradation, and to efficiently transfer the system to the 2nd–stage, with the purpose of achieving the remediation goal and improving the problem of the residual concentrations.
The soil contaminated by 14,032 mg total petroleum hydrocarbon (TPH) /kg dry soil was employed in the study to find an optimal bioremediation approach. The bioremediation approach of combined bioaugmentation and biosurfactant, with introduction of five strains bacteria of 108 CFU/g dry soil and 100 mg/kg soil of rhamnolipid (A8R100), reached the first stage of degradation in 35 days and removal efficiency of 70%. The problem of slow TPH degradation found to be related to the polar fractions accumulation. Increase of fungi community diversity was found helpful in decomposition of the polar fractions. The symbiotic environment between bacteria and fungi was beneficial to the 2nd –stage TPH biodegradation.
Based on the above findings, we designed a two-stage remediation strategy, by using the A8R100 approach during the 1st stage. Then, when TPH was persistent to be further degraded, we enhanced the fungi diversity in the remediation system in order to shorten the lag phase and to improve the degradability particularly for the polar substances. Our result indicated that, with the A8R100 approach, the soil initially contaminated with 7,379 mg TPH/kg dry soil could complete the 1st – stage degradation by the 1st 21 days, and the removal efficiency was 65%. We then added hydrocarbon degrading fungi and yeast in the second-stage, and the removal efficiency was raised to be 87% and 89%, respectively. When the compost kitchen waste (KW) strategy was employed, the TPH removal efficiency was higher than 89%.
The two-stage bioremediation strategy could help us to attain the Taiwan Soil and Groundwater Remediation Regulation, TPH lower than 1,000 mg/kg dry soil, within three months. The application of fungi and kitchen waste found to be useful in shorten the lag phase during the 2nd-stage degradation and achieve the remediation goal efficiently.

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