Characterization of Denitrifying Phosphorus Removal Microorganisms in a Novel Two-Sludge Process by Combining Chemical with Microbial Analysis

The present work focuses on the investigation of denitrifying phosphorus removal organisms (DPB) in a novel two-sludge denitrifying phosphorus removal process by combining chemical with microbial analysis. When the two-sludge process operated stably over one year, good phosphorus (P) release and P uptake performance of activated sludge samples collected from this process were present in anaerobic and anoxic conditions, respectively, via batch test, showing that the ratio of P release specific rate to P uptake specific rate was 1.31. The analysis of energy dispersive spectrometry (EDS) showed that P content of activated sludge samples collected at the end of anoxic phase was 12.3% of dry weight, further demonstrating the existence of microorganisms responsible for phosphorus removal in this two-sludge process. From polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) analysis, the presence of microorganisms mostly belonging to the phyla Firmicutes and Proteobacteria was observed, previously evidenced in the phosphorus removal wastewater treatment process. Fluorescence in situ hybridization (FISH) quantitative analysis showed that Accumulibacter responsible for phosphorus removal was dominant in this two-sludge process, accounting for 69.7% of all bacteria in activated sludge. These results obtained from chemical and microbial analysis in this study suggested that denitrifying phosphorus removal microorganisms were completely enriched in the two-sludge process proposed here.