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年度	112	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	大仁科技大學	申請系所	環境與職業安全衛生系
專案主持人	賴文亮	職等/職稱	教授
專案中文名稱	新穎蟲生整治技術處理受重質石油碳氫化合物污染土壤
中文關鍵字	石油碳氫化合物, 黑水虻, 綠色土壤整治技術
專案英文名稱	Novel Insect-Mediated Remediation Technique to remediate Heavy Petroleum Hydrocarbon contaminated soils
英文關鍵字	Total petroleum hydrocarbons, Black solider fly, Green Soil remediation
執行金額	1,079,000元
執行期間	2024/1/1 至 2024/12/31
計畫中文摘要	本本研究團隊選用三齡黑水虻幼蟲(Black soldier fly larvae, BSFL) 進行人工配製受柴油與燃料油污染土壤，總石油碳氫化合物(Total petroleum hydrocarbons,TPH)劑量介於2,000-10,000 mg/kg，土樣1公斤加10%米糠，投入適量三齡BSFL幼蟲數，進行BSFL在兩種油品耐受力、生長速率、BSFL對低碳數(C6-C9)及高碳數(C10-C50)TPH之總去除率、土壤肥力評估及土壤與BSFL菌相豐富度變化進行探討，作為未來綠色工法整治之參考。 研究成果區分四部分，說明如下:(1)在蟲體耐受性及安全性:BSFL於燃料油之死亡率及預蛹率，均低於同劑量柴油之值。BSFL幼蟲存活比例於受油品污染土壤，超過90%，代表其在油品之耐受程度相當高。 BSFL經處理柴油及燃料油後，蟲體內並無檢測到油品標準中高低碳數TPH成份之累積。(2) 減量評估: BSFL對柴油與燃料油中TPHlow-carbon去除應可有效控制(6,000-10,000 mg/kg)，但在燃料油低劑量(2,000mg/kg) ，需注意BSFL代謝釋出TPHlow-carbon之干擾。BSFL對燃料油中TPHhigh-carbon去除率表現於柴油更佳。BSFL對TPH之生物質轉換速率，在低碳數部分，燃料油高於柴油；在高碳數部分，則是柴油高於燃料油。油品劑量落於4,000-8,000 mg/kg，BSFL在燃料油之生長速率明顯高於在柴油之表現。(3)減污土壤肥力測試:不同油品劑量土壤，經BSFL處理後之pH值、導電度、有機質、有效氮含量，均高於對照組，且Amino(I+II+VI) acid/Humic(III+V)螢光強度比值，隨處理時間之增加而增加，兩油品呈現一致趨勢。油品萃出液對小麥發芽率之影響，燃料油之表現優於柴油，兩種油品對小麥發芽率均優於對照組，但對玉米及水稻發芽率，兩種油品則低於對照組。(4)菌種鑑定: 土壤中菌相豐富度分析中，對照組與實驗組雖有所變化，可觀察Dysgonomonas對兩種油品高低劑量燃料之耐受力高；Brevibacterium 菌屬，在兩種油品劑量增加時，其菌相豐富度隨之增加，BSFL腸道中菌相Rhodococcus (紅球菌屬)實驗組(添加柴油或是燃料油)豐富度比對照組多出10倍以上。利用QIIME中的Sorensen dice index analysis統計方法, 對照組與含柴油或是燃料油之土相或BSFL腸道菌相具有差異性。本研究初步驗證BSFL應用於受油品污染場址之可行性，但受限各項資源，仍有需完善補強不足之處，進而作為綠色整治開發技術及現場應用之參考。
計畫英文摘要	In this study, we selected third-instar Black Soldier Fly Larvae (BSFL) to treat artificially contaminated soil with diesel and fuel oil, with total petroleum hydrocarbons (TPH) concentrations ranging from 2,000 to 10,000 mg/kg. One kilogram of soil was mixed with 10% rice bran and inoculated with an appropriate number of third-instar BSFL to evaluate their tolerance to both oils, growth rates, total removal rates of low-carbon (C6-C9) and high-carbon (C10-C50) TPH, changes in soil fertility, and microbial diversity in both soil and BSFL. These findings are intended as a reference for future green remediation technologies. The study is divided into four parts:(1) Larval Tolerance and Safety: BSFL mortality and pre-pupation rates in fuel oil were lower than those in diesel at the same TPH levels. The survival rate of BSFL in oil-contaminated soil exceeded 90%, indicating a high tolerance to both oil types. No accumulation of TPH components (low or high carbon number) was detected in the larvae after exposure to diesel or fuel oil. (2) TPH Reduction Assessment: BSFL should effectively control TPHlow-carbon removal (6,000-10,000 mg/kg) from diesel and fuel oil, but at low doses in fuel oil (2,000 mg/kg), attention should be paid to potential interference from BSFL metabolic release of TPHlow-carbon. BSFL demonstrates better TPH high-carbon removal efficiency in fuel oil than diesel. Regarding the biomass conversion rate of TPH, in the low-carbon fraction, fuel oil surpasses diesel; in the high-carbon fraction, diesel exceeds fuel oil. For TPH levels between 4,000 and 8,000 mg/kg, BSFL exhibited significantly higher growth rates in fuel oil, likely due to the more straightforward conversion of low-carbon TPH in fuel oil. (3) Soil Fertility Tests: Soil treated with BSFL showed higher pH, electrical conductivity, organic matter, and available nitrogen content than the control group. The fluorescence intensity ratio of Amino acids (I+II+VI) to Humic acids(III+V) increased over time, with both oil types showing a consistent trend. Wheat germination rates in the oil-contaminated extracts were higher for fuel oil than diesel. Both oils improved wheat germination compared to the control, though germination rates for corn and rice were lower than the control. (4) Microbial Identification: Microbial diversity analysis of soil revealed changes between the control and experimental groups. Dysgonomonas showed high tolerance to both oils at various concentrations, while the abundance of Brevibacterium increased with higher oil concentrations. In the BSFL gut microbiome, the abundance of Rhodococcus was over ten times higher in the oil-treated groups compared to the control. Statistical analysis using the Sorensen Dice Index in QIIME showed significant differences in microbial diversity between the control and oil-treated groups in both soil and BSFL guts. This study preliminarily confirms the feasibility of using BSFL to remediate oil-contaminated sites. However, resource limitations require further improvement before these findings can be applied as green remediation technologies for field applications.