Interrogating the performance and microbial ecology of an enhanced biological phosphorus removal/post-anoxic denitrification process at bench and pilot scales.

Research focused on interrogating post-anoxic enhanced biological phosphorus removal (EBPR) at bench and pilot scales. Average bench-scale effluent ranged from 0.33 to 1.4 mgP/L, 0.35 to 3.7 mgNH3 -N/L, and 1.1 to 3.9 mgNOx -N/L. Comparatively, the pilot achieved effluent (50th percentile/average) of 0.13/0.2 mgP/L, 9.7/8.2 mgNH3 -N/L, and 0.38/3.3 mgNOx -N/L under dynamic influent and environmental conditions. For EBPR process monitoring, P:C ratio data indicated that 0.2-0.4 molP/molC will result in stable EBPR; relatedly, a target design influent volatile fatty acid (VFA):P ratio would exceed 15 mgCOD/mgP. Post-anoxic EBPR was enriched for Nitrobacter spp. at 1.70%-20.27%, with Parcubacteria also dominating; the former is putatively associated with nitritation and the latter is a putative fermenting heterotrophic organism. Post-anoxic specific denitrification rates (SDNRs) (20°C) ranged from 0.70 to 3.10 mgN/gVSS/h; there was a strong correlation (R2 = 0.94) between the SDNR and %Parcubacteria for systems operated at a 20-day solids residence time (SRT). These results suggest that carbon substrate potentially generated by this putative fermenter may enhance post-anoxic EBPR. PRACTITIONER POINTS: Post-anoxic EBPR can achieve effluent of <0.2 mgP/L and <12 mgN/L. The P:C and VFA:P ratios can be predictive for EBPR process monitoring. Post-anoxic EBPR was enriched for Nitrobacter spp. over Nitrospira spp. and also for Parcubacteria, which is a putative fermenting heterotrophic organism. Post-anoxic specific denitrification rates (20°C) ranged from 0.70 to 3.10 mgN/gVSS/h. BLASTn analysis of 16S rDNA PAO primer set was shown to be improved to 93.8% for Ca. Accumulibacter phosphatis and 73.2%-94.0% for all potential PAOs.

Interrogating nitritation at a molecular level: Understanding the potential influence of Nitrobacter spp.

Water resource recovery facilities (WRRFs) increasingly must maximize nitrogen and phosphorus removal, but concurrently face challenges to reduce their energy usage and environmental footprint. In particular, biological nutrient removal (BNR), which targets removal of phosphorus and nitrogen, exhibits a large energy demand. However, a BNR process achieving partial oxidation of NH3 to NO2 (nitritation) could reduce energy demands, with secondary environmental emission benefits. Research was conducted on bench-scale systems performing nitritation and nitrification to better understand how mixed microbial consortia, cultured on real wastewater, can sustain nitritation. BNR configurations achieved nitrite accumulation ratios of 64-82%, with excellent overall effluent quality. Applying phylogenetic, transcriptomic, and metabolomic methods, coupled with process monitoring, results indicate that partial nitritation may be induced through a combination of: (1) Employing ammonia-based aeration control, with an ammonia setpoint of 2, 3 mgN/L; (2) Maintaining an aerobic period DO of 1.0-2.0 mg/L; and (3) Operating BNR post-anoxically, integrated within enhanced biological phosphorus removal (EBPR). Significant nitritation was achieved despite the presence Nitrobacter spp., but nitrite oxidoreductase must be functionally impaired or structurally incomplete. Overall, this research demonstrated the value of interrogating a mixed microbial consortia at a macro and molecular level to explore unique metabolic responses such as nitritation.