反硝化聚磷菌快速富集、培养及其荧光原位杂交技术鉴别

Rapid Enrichment and Cultivation of Denitrifying Phosphate-Removal Bacteria and Its Identification by Fluorescence in situ Hybridization Technology

以污水处理厂A2/O厌氧段污泥为种泥,采用膜生物反应器(membrane bio-reactor,MBR)对反硝化聚磷菌(denitrifyingphosphate-removal bacteria,DPB)进行快速富集及培养,并提供一种鉴别方法.试验中以乙酸钠为碳源,并在缺氧段投加一定浓度的硝酸盐,结果表明,在膜组件的高效截留作用下,经过厌氧-好氧和厌氧-缺氧2个阶段的富集培养,35 d内反硝化聚磷菌占全部聚磷菌(phosphate-accumulating organisms)的比例从24%上升到93%.此时系统的脱氮、除磷效率均可保持在90%以上.通过荧光原位杂交技术(fluorescence in situ hybridization,FISH)并结合常规测定手段对活性污泥进行鉴别,确定Pseudomonas sp.和Rhodocyclus sp.为主要的优势菌.

The present work focused on a rapid enrichment and cultivation of denitrifying phosphate-removal bacteria（DPB）in a membrane bio-reactor（MBR） by using A2/O anaerobic sludge from a wastewater treatment plant as seed, as well as providing an identification method. In the experiments, sodium acetate was used as the carbon source and a certain amount of nitrate was added to the MBR in the anoxic stage. Results showed that, with the efficient trap of the hollow-fiber membrane module, the proportion of DPB in all the phosphate-accumulating organisms （PAOs） increased from 24% to 93% within 35 days after two-stage＇s cultivation including anaerobic/aerobic and anaerobic/anoxic, during which the removal efficiency of nitrogen and phosphorus reached more than 90%. The activated sludge was identified by combining a regular method and the fluorescence in situ hybridization （FISH） technique, which demonstrated that Pseudomonas sp. and Rhodocyclus sp. were the dominant bacteria in the used bioreactor.