利用分子生物技術分析光電業廢水生物處理單元菌相結構之研究

Molecular biological analysis of microbial community in biological treatment of photoelectrical industry wastewater

林亦識(I-Shih Lin)

指導教授 ： 曾迪華

國立中央大學/工學院/環境工程研究所/碩士(2006年)

摘要

本研究使用不同的16S rRNA族群引子，搭配PCR-DGGE技術，分析實廠處理光電業廢水之Barnard程序中的菌相結構，以建立相關菌種資料。實驗結果顯示，不同族群引子，包括Bacteria、Kingdom Acidobacterium、α-Proteobacteria、β-Proteobacteria Ammonia Oxidizers、Actinomycetes及Methylotrophs等族群，搭配PCR-DGGE實驗，不僅可在選殖文庫進行前，快速獲得微生物族群分類的資訊，且有助於少數族群菌相結構的觀察。此外，PCR-DGGE分析的結果，可做為選殖文庫設計的參考，提高選殖效率。本研究樣本來自於實廠處理光電業廢水的Barnard程序，缺氧-好氧-缺氧-好氧生物處理槽的懸浮污泥。16S rRNA選殖文庫由Bacteria和β-Proteobacteria Ammonia Oxidizers的PCR產物所建立，分別篩選出11株與8株選殖株。菌相結構分析結果顯示，β-Proteobacteria為本研究之優勢族群，有18株屬於此族群，而有6株選殖屬於草酸桿菌科(Oxalobacteraceae)。Janthinobacterium sp.為最主要的優勢菌種，各槽皆有超過50%的相對數量；Nitrosospira sp.為β-Proteobacteria Ammonia Oxidizers族群中的優勢菌種，此族群於四個槽體中的相對數量皆超過30%。選殖編號B3、N2、N5之菌株經基因資料庫比對後發現，與資料庫已知序列之相似度皆小於97%，推測此三株菌可能為本研究所發現的新菌種。

關鍵字

族群引子 ； 菌相結構 ； PCR-DGGE ； 光電業廢水 ； 銨氧化菌

並列摘要

The microbial community structure analysis of a full-scale Barnard process for treating photoelectrical industry wastewater was investigated in this study. The group-specific 16S rRNA primers and PCR-DGGE method were applied to figure out the profile of the microbial communities and the connection to the processes. Experimental results indicate that with different group-specific 16S rRNA primers including bacteria, kingdom acidobacterium, α-proteobacteria, β-proteobacteria ammonia oxidizers, actinomycetes, and methylotrophs, coordinate to PCR-DGGE method, can facilitate the classification of dominant species before clone library establish and promote the sensitivity of the PCR-DGGE method. In addition, the results of PCR-DGGE analyzing can be a good reference for the clone library design which enhances the efficiency of the cloning for the followed experiments. Suspending sludge samples for this study were collected from a full-scale Barnard process by treating the photoelectrical industry wastewater. The process is constituted by an anoxic tank-aerobic tank-anoxic tank-aerobic tank treatment schedule. The 16S rRNA clone library was established by PCR product from bacteria and β-proteobacteria ammonia oxidizers, which including 11 bacteria and 8 β-proteobacteria ammonia oxidizers clones. Microbial community analysis result shown that the β-proteobacteria is the dominant species in all the Barnard process biological processes. Furthermore, in the clone library, 18 clones belonged to β-proteobacteria and 6 of them demonstrate similarity to Oxalobacteraceae. Janthinobacterium sp. is the mainly dominant species and distributed more than 50% of bar weight to the total population. Nitrosospira sp. is also a dominant species in the β-proteobacteria ammonia oxidizers with distribution of more than 30% of the total population in the system. The bacterial 16S rRNA gene sequences of clone B3, N2, and N5 shown less than 97% similarity to the known strain in the database of NCBI. Accordingly, the clone B3, N2, and N5 obtain the probability for belong to the novel discovery strains which found by this study.

並列關鍵字

Microbial community ； photoelectrical industry wastewater ； ammonium oxidizers ； PCR-DGGE ； group-specific primers

參考文獻

陳雯怡，「活性污泥銨氧化基因多樣性之研究」，碩士論文，中原大學土木工程學系，中壢(2005)。

財團法人光電科技工業協進會(2006)，「全球光電市場與台灣光電產業回顧與展望」，http://www.pida.org.tw/welcome.asp

袁熙隆、蔡孟謙、施健堃、黃良銘，「都巿與畜牧廢水生物處理系統硝化菌生態與硝化效能相關性之研究」，第十六屆環工年會研討會論文集，臺南(2004)。

楊雅斐，「三段生物程序中好氧硝化槽功能評估與分生檢測生態研究」，碩士論文，國立成功大學環境工程學系，臺南(2003)。

申忠哲，「處理煉焦廢水之活性污泥槽中硫氰酸分解菌的族群結構之研究」，碩士論文，國立成功大學生物學研究所，臺南(2004)。