PCR-DGGE法研究Sludge bio-membrane(SB)系统中反硝化聚磷菌的变化

摘要

Sludge bio-membrane nitrogen and phosphorus removal system was used to enrich denitrifying phosphorus bacteria(DPB).Plate separation and PCR-DGGE technology were applied to monitor the diversity of bacteria.The results show that before the enrichment, isolated micro-organisms are mainly seven kinds of bacteria including Corynebacterium sp., Acinetobacter sp., Pseudomonas sp., Enterobacteriaceae sp., Moraxella sp., Staphylococcus sp.and Paracoccus sp.,seven kinds of bacteria are isolated by the plate separation.After enrichment four kinds of main bacteria i.e.,Acinetobacter sp., Pseudomonas sp., Enterobacteriaceae sp., and Paracoccus sp., are found.The species of bacteria are reduced after enrichment and DPB in SB system are different in the SBR.Using PCR-DGGE to investigate bacteria,Flavobacterium sp., Alcaligenes sp., Paracoccus sp., Janthinobacterium sp.and Erythrobacter sp., are mainly five kinds of bacteria before enrichment, but only three kinds of bacteria i.e., Alcaligenes sp., Paracoccus sp., and Janthinobacterium sp.are remained after enrichment.Paracoccus sp.is the only DPB which is confirmed by two methods.Proteobacteria sp.dominates the denitrifying phosphorus bacteria when PCR-DGGE and 16S rDNA clone method are used to analyze the diversity of bacteria.%采用SB同步脱氮除磷系统富集反硝化聚磷菌,利用平板分离法和PCR-DGGE技术,进行微生物种群的跟踪.研究结果表明:通过平板法分离到的微生物主要为棒状杆菌属、不动杆菌属、假单胞菌属、肠杆菌科、莫拉氏菌属、葡萄球菌属、副球菌属共7种,富集后细菌主要为不动杆菌属、假单胞菌属、肠杆菌科、副球菌属4种,富集后系统内细菌种类减少,与采用Sequencing Batch Reactor(SBR)池富集的反硝化聚磷菌不同.采用PCR-DGGE法发现富集前以黄杆菌属、产碱杆菌属、副球菌属、紫色杆菌属、赤细菌属为主,富集后以产碱杆菌属、副球菌属、紫色杆菌属为主,副球菌属是唯一通过2种办法获得确认的反硝化聚磷菌株.采用PCR-DGGE和16S rDNA克隆文库方法研究的同步反硝化聚磷菌都以变形门占优势.