本研究採用系統環境分子生物復育技術以促進兩階段生物復育法的效能，利用添加廚餘廢棄物與木屑混拌腐熟堆肥至受石油碳氫化合物汙染的土壤中是項新穎的生物處理方法，廚餘腐熟堆肥的添加除了作為土質改良劑外、當中的真菌族群亦作為生物添加以促使石油碳氫化合物的生物降解。實驗用的土壤採樣自煉油廠汙染場址，依受污染土壤中總碳氫化合物的碳數和分子量可分成低碳數的砂質壤土(LC)、高碳數的粉質黏土(HC)、以及兩者混和後的綜合性碳數土壤(MC)。實驗室前人研究中探討在不同總有機碳下添加廚餘堆肥對總石油碳氫化合物的降解情形，證實有助於降解低碳數部份。在第一階段，本研究分別添加1.5%和3%廚餘堆肥(w/w)到不同土壤中，判定省時又能降低成本的最佳操作條件。

第一階段的結果顯示添加廚餘堆肥不但提高了降解效率、微生物族群的多樣性也有所提升(包含總異營菌-THB、碳氫化合物降解菌-HDB、以及真菌群)。添加3%廚餘堆肥的低碳數土壤反應槽(LC-2)有最佳的降解表現，降解率高達77%；然而高碳數以及混和的綜合碳數反應槽(HC, MC)降解速率則呈現緩慢遲滯。結果顯示和粉質黏土壤相比，微生物生長較適應於砂質壤土的環境下、總石油碳氫化合物也比較容易被微生物所降解。

為了促進先前土壤反應槽中高分子量碳氫化合物的降解效率，在第二階段中，另外加入了五種已知可降解石油碳氫化合物的真菌族群。而結果亦顯示添加真菌可以促進生物復育的效率，尤其是砂質壤土的環境下。真菌群的添加也造就THB、HDB、以及總異營真菌群THF數量上的成長，然而真菌族群往往在加入反應槽後的20-40天內衰減，推測為細菌和真菌群競爭碳源(石油碳氫化合物)的消長結果。添加真菌群的目的是要降解石油碳氫化合物中的芳香族、resin和極性物質，但對於高碳數及混和綜合碳數反應槽依然無法有效降解之。在第二階段後，混和的綜合碳數土壤反應槽(MC-2)的總石油碳氫化合物降解率僅65%、高碳數反應槽(HC-2)降解率為72%、反觀低碳數反應槽(LC-1,LC-2)則有97%的降解率。針對上述結果另外做一實驗，目標為最佳化粉質黏土下受新鮮柴油汙染的降解率表現，最佳化結果為添加5%含真菌群的廚餘堆肥(KWF)有最高的降解效率，在63天操作下就有91%的降解效率。

利用分子生物偵測技術的結果，可以確認原生的油降解菌和真菌。在LC, MC, HC反應槽中的主要優勢細菌為Sphingomonas sp. ；在第三階段的CT, KW, F, KWF反應槽則是以Acinetobacter junii 為優勢種；三個階段中皆以Candida 和 Fusarium為主要優勢真菌群。當廚餘堆肥或真菌族群加入反應槽中時，土壤菌相會改變，但菌相多樣性則會隨碳源的消耗而減少。

Two phases of bioremediation technologies, Kitchen waste (KW) compost amendments and bioaugmentation by adding fungi consortium were a part of an innovative bioprocess method, Systematic Environmental Molecular Bioremediation Technology (SEMBT) which developed as an integrated bioremediation technology in order to enhance the Total Petroleum Hydrocarbon (TPH) degradation from aged TPH contaminated soil. The contaminated soil were taken from an oil production plant exposed with contaminant for a long time and divided into Low Carbon (LC) with loamy sand texture contaminated with diesel oil (C10~C28), High Carbon (HC) with silty clay texture contaminated with fuel oil (C10~C40) and combination between LC and HC called Mixed Carbon (MC) based on their molecular weight carbon number. Previous study in our lab investigated the TPH degradation using KW compost with different levels of Total Organic Carbon (TOC). In present study 1.5% and 3% of KW compost (w/w) were applied in different aged TPH contaminated soil at batch 1 (day 0-77) to determine the optimal strategy with time saving and cost effective.

The result from batch 1 (Day 0-77) study showed that adding KW compost enhanced not only the degradation efficiency but also the microbial population intensively (the Total Heterotrophic bacteria (THB), Hydrocarbon Degrading Bacteria (HDB) and Fungi). The best degradation performance achieved was 77% in LC-2 bioreactors with 3% of KW compost amendment. Slow degradation was detected on High Carbon and Mixed Carbon bioreactors respectively. Loamy sand texture provided more favorable conditions for the growth of microorganisms capable to degrade TPH than the silty clay texture. The loamy sand texture also made contaminant (TPH) more accessible and available to be degraded by microorganisms rather than silty clay texture.

In order to enhance the degradation of high molecular weight hydrocarbon in previous soil bioreactors, five strains of fungi consortium were applied at batch 1 (Day 77-192). The result showed that adding fungi enhanced bioremediation efficiency in the aged TPH contaminated soil especially in loamy sand soil texture. Fungi consortium amendment also enhanced the THB, HDB and also THF population growth. But Fungi growth easily depleted within approximately 20-40 days after they were introduced into the bioreactors because there was a competition between bacteria and fungi to utilize the contaminant as their carbon sources. The purpose of adding fungi consortium to degrade the aromatic, resin and polar compounds was not very successful applying in Mixed Carbon and High Carbon bioreactors the best TPH degradation was 65% in MC-2 bioreactor and 72% in HC-2 bioreactor whilst 97% TPH degradation detected in LC-1 and LC-2 bioreactors.

Another study being conducted to find the optimal strategy of TPH degradation in fresh diesel oil contaminated soil with silty clay texture, the best bioremediation performance was 91% during 63 days study could be achieved if we combined 5% of KW compost with fungi consortium (KWF).

The result of molecular biomonitoring confirmed the existence of indigenous oil degrading bacteria and fungi. Sphingomonas sp. was detected as the dominant bacteria existed in LC, MC and HC series bioreactors whilst in batch 2 study, Acinetobacter junii was dominant in CT,KW,F and KWF bioreactors. Candida and Fusarium were two dominant genus existed in all bioreactors during batch 1 and 2 study. In particular when KW compost and fungi consortium were introduced in bioreactors, the diversity of bacteria and fungi become more diverse but their diversity will become less when their carbon sources were consumed.

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