Contribution of Pseudomonas spp. to phosphorus uptake in the anoxic zone of an anaerobic-anoxic-aerobic continuous activated sludge system

Abstract

A continuously operated laboratory-scale (32 L) nitrification denitrification biological excess phosphorus removal (NDBEPR) activated sludge system (modeled on the 3-stage Phoredox configuration) was maintained for 140 d. The transition from a non-biological excess phosphorus removal (BEPR) sludge to one exhibiting a strong BEPR mechanism was monitored. Mixed liquor seed inoculum was obtained from a full-scale single aerobic activated sludge installation and subjected to conditions conducive to BEPR, i.e. increasing influent acetate (HAc) concentrations. At a sludge age of 10 d with 100% HAc feed, the system was capable of removing a maximum of ca. 40 mgPO4–P/L from the bulk liquid; P/VSS of ca. 0.27 (mgP/mgVSS); and VSS/TSS of 0.53 (mgVSS/mgTSS) in the aerobic zone was attained. Although typical BEPR phosphorus transformation patterns were routinely observed, i.e. anaerobic phosphate release and aerobic phosphate uptake, phosphate uptake in the anoxic zone was also recorded indicating the presence of denitrifying phosphorus accumulating organisms (DPAOs) in the sludge community. The microbial community was screened (using both isolation and direct methods of analysis) for the presence of Pseudomonas spp. as this genus is known to perform both polyphosphate accumulation and denitrification processes. Isolation of anoxic mixed liquor bacteria on solid media and identification using the API 20NE system resulted in the total dominance of the Pseudomonads (>50%). However, direct fluorescent in situ hybridizations (FISH) revealed that Pseudomonas spp. only constituted ca. 3% of the total bacterial community indicating that other bacterial genera are contributing to simultaneous polyphosphate accumulation and denitrification processes in the anoxic zones of NDBEPR systems.