近年來台灣的土壤及地下水受油品污染的情況日益嚴重，其中儲油槽與管路之老舊，所造成的管線破裂、洩漏等是其主因。在汰舊換新之際的環境復原，當今環保科技公司常以物理或化學性的處理來解決，然二次環境污染之虞卻又不可免。在此窘境之中，自古既有，然效果較為緩慢的生物復育技術才又被重視。此技術乃將油分解菌佈入土壤中來進行，然迥異於以往者，現今技術著重於提高土壤復育的時效性。依此為前提之中，除搭配絕佳的菌劑與恰當的助劑來佈施之外，能夠迅速反應出土壤內油分解菌活性的監測技術實不可缺。本研究之目的，旨在希望研發出能鑑定土壤內的油分解菌之生物介電泳(DEP)晶片的分離技術平台，繼而結合拉曼光譜法對菌株進行鑑定，使本研究團隊針對重油與柴油之分解所選擇的E. aurantiacum CC-LSH4-1和A. junii CC-FH2以及 R. erythropolis CC-BC11和G. alkanivorans CC-JG39，配合生物性界面活性劑進行施工後的效果得以被評估。本研究利用上述四株菌不同介電特性設計出擁有平行式電極、箭頭型電極和城垛式電極等三種設計的三維DEP晶片，於稀釋的PBS溶液(2 mS/cm)下成功地同時分離此四株菌。DEP晶片的交流電條件方面，若設予Vp-p為15 V，而頻率各為4、8、15 MHz時，會顯示出正介電泳的作用力，於前三處電極區，可分別捕捉到E. aurantiacum CC-LSH4-1, A. junii CC-FH2及R. erythropolis CC-BC11，而G. alkanivorans CC-JG39則在最後的電極區以300 kHz，呈負介電泳力作用方式被捕捉到。另在拉曼光譜分析上，發現在633 nm氦氖雷射(17 mW)下，此四株菌均有其特異光譜特性，我們擷取其各波數與強度值，導入於主成份分析法(PCA)和階層聚類分析法(HCA)進行菌種的分類，發現已可明顯將此四株菌的光譜分別歸類，這兩種資料建立將可縮短光譜的分析時間並降低人為判斷的誤差。繼之也以G. alkanivorans CC-JG39為例，先經DEP分離後釋出取樣，此樣本液置於鍍金玻片上再經拉曼光譜分析，從PCA與HCA辨認出為該細菌無誤的結果。最後，我們雖然曾嘗試以懸浮搭配離心之前處理，想從含菌之土壤樣本中取得其細菌樣本，唯所含的極細微粒會有堵住微管道之虞，且前處理後的菌相與菌量，無法確保，導致較具體的結果尚待檢討，但本技術平台未來應有機會被運用在土壤內油分解菌活性的監測上。

Recently, with industry accidents like leaking oil from old pipe, oil container, the problems of oil pollution in soil and groundwater are getting more and more seriously. We use bioaugmentation method of bioremediation technologies to execute soil bioremediation. And then we choose Exiguobacterium aurantiacum CC-LSH4-1, Acinetobacter junii CC-FH2, Rhodococcus erythropolis CC-BC11 and Gordonia alkanivorans CC-JG39 according to their high degradation rate. The purpose of our research is to combine DEP chip and Raman spectroscopy to separate and identify four kinds of oil degradation bacteria from the soil. The 3D DEP chip was exclusively made according to the dielectric properties of four kinds of oil degradation bacteria. Consequently, we design three kinds of electrode which are parallel, arrow and castellated electrode. In the PBS (2 mS/cm) solution, E. aurantiacum CC-LSH4-1, A. junii CC-FH2 and R. erythropolis CC-BC11 were individually trapped at 4 MHz, 8 MHz and 15 MHz by positive DEP . At the same time, G. alkanivorans CC-JG39 was trapped at 300 kHz by negative DEP. After that we can find 30% of similarity and 70% of dissimilarity from qualitative analysis results according to four kinds of bacteria wavenumber. And then we also use principal component analysis and hierarchical cluster analysis methods to execute classification of bacteria. The four kinds of bacteria can be categorized in different area. Currently with this DEP chip we can accomplish the experiment of the trapping for oil degradation bacteria in the soil. by Spectra Analysis,qualitative analysis,statistic analysis we can discriminate the difference of these bacteria in Raman spectra, and therefore this device will apply to the examination of these oil degradation bacteria from soil in the future.

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