Synergistic Integration of Anammox and Endogenous Denitrification Processes for the Simultaneous Carbon, Nitrogen, and Phosphorus Removal.

The feasibility of a synergistic endogenous partial denitrification-phosphorus removal coupled anammox (SEPD-PR/A) system was investigated in a modified anaerobic baffled reactor (mABR) for synchronous carbon, nitrogen, and phosphorus removal. The mABR comprising four identical compartments (i.e., C1-C4) was inoculated with precultured denitrifying glycogen-accumulating organisms (DGAOs), denitrifying polyphosphate-accumulating organisms, and anammox bacteria. After 136 days of operation, the chemical oxygen demand (COD), total nitrogen, and phosphorus removal efficiencies reached 88.6 ± 1.0, 97.2 ± 1.5, and 89.1 ± 4.2%, respectively. Network-based analysis revealed that the biofilmed community demonstrated stable nutrient removal performance under oligotrophic conditions in C4. The metagenome-assembled genomes (MAGs) such as MAG106, MAG127, MAG52, and MAG37 annotated as denitrifying phosphorus-accumulating organisms (DPAOs) and MAG146 as a DGAO were dominated in C1 and C2 and contributed to 89.2% of COD consumption. MAG54 and MAG16 annotated as Candidatus_Brocadia (total relative abundance of 16.5% in C3 and 4.3% in C4) were responsible for 74.4% of the total nitrogen removal through the anammox-mediated pathway. Functional gene analysis based on metagenomic sequencing confirmed that different compartments of the mABR were capable of performing distinct functions with specific advantageous microbial groups, facilitating targeted nutrient removal. Additionally, under oligotrophic conditions, the activity of the anammox bacteria-related genes of hzs was higher compared to that of hdh. Thus, an innovative method for the treatment of low-strength municipal and nitrate-containing wastewaters without aeration was presented, mediated by an anammox process with less land area and excellent quality effluent.

Unravelling the importance of Ca2+ and Mg2+ as essential in anammox culture medium.

The mechanism and nitrogen removal performance of anammox process under different concentrations of Ca2+ and Mg2+ were explored from the perspective of molecular biology analysis based on the metabolic changes of the second messenger cyclic diguanylate (c-di-GMP). After 100-day operation, reactor with 98 mg/L Ca2+ and 30 mg/L Mg2+ achieved a higher anammox performance with an average total nitrogen (TN) removal efficiency of 85.8%. Under the Mg2+concentration of 30 mg/L, a higher Ca2+ could accelerate anammox process by promoting the amplification of Candidatus Brocadia (0.62%) and production of Diguanylate cyclase (DGC-s: 6.54 × 108 copies/µL DNA) which function was to synthesize c-di-GMP. While under the Ca2+concentration of 49 mg/L, Mg2+ concentration at appropriate rang could promote the degradation process of c-di-GMP. Since Ca2+ had positive linear relationship with TN removal (R2 = 0.96), a higher Ca2+ concentration is recommended in the culture medium. This study provided a potential method for optimization of anammox process.

Achieving simultaneous nitritation, anammox and denitrification (SNAD) in an integrated fixed-biofilm activated sludge (IFAS) reactor: Quickly culturing self-generated anammox bacteria.

In this study, by inoculating nitritation suspended sludge, simultaneous nitritation, anammox and denitrification (SNAD) was established quickly in an integrated fixed-biofilm activated sludge (IFAS) reactor to treat high-ammonia municipal wastewater. Results showed that, deep-level total nitrogen and chemical oxygen demand removal efficiencies (92.8% and 78.8%, respectively) were achieved, and their effluent concentrations were 13.2 and 39.3 mg/L, respectively. Excess generation of nitrate was once occurred under continuous aerobic condition, but it could be solved by suppressing nitrite oxidizing bacteria activity stably via switching to intermittent aeration mode (alternate 7 min of aerobic and 21 min of anoxic) and rising influent ammonium concentration temporarily (lasted 31 days). High-throughput sequencing analysis revealed that, Candidatus_Brocadia, as dominant anammox bacteria, was self-generated in flocs (2.93%) but mainly biofilm (7.67%), whereas uncultured_f_Nitrosomonadaceae as ammonia oxidizing bacteria was mainly found in flocs (2.4%). This work not only demonstrated that anammox bacteria could be self-generated and retained in the SNAD-IFAS system, but also suggested a promising application of the SNAD-IFAS in wastewater treatment plants.

Achieving deep-level nutrient removal via combined denitrifying phosphorus removal and simultaneous partial nitrification-endogenous denitrification process in a single-sludge sequencing batch reactor.

The feasibility of coupling denitrifying phosphorus removal (DPR) with simultaneous partial nitrification-endogenous denitrification (SPNED) was investigated in a single-sludge sequencing batch reactor for deep-level nutrient removal from municipal and nitrate wastewaters. After 160-day operation, the DPR process simultaneously reduced most PO43--P and NO3--N anoxically, and the SPNED process achieved further total nitrogen (TN) removal at low dissolved oxygen condition with TN removal efficiency of 90.8%. The effluent NH4+-N, PO43--P and TN concentrations were 1.0, 0.1 and 7.2 mg/L, respectively. Microbial analysis revealed that Dechloromonas (6.7%) dominated DPR process, whereas the gradually enriched Nitrosomonas (4.5%) and Candidatus Competibacter (6.8%) conducted SPNED process accompanied with sharply eliminated Nitrospirae (1.4%). Based on these findings, a novel strategy was proposed to achieve further nutrient removal in conventional treatment through integrating the DPR-SPNED process. As a result, ∼100% of extra carbon and ∼10% of oxygen consumptions would be reduced with satisfactory effluent quality.

[Nitrogen and Phosphorus Removal from Low C/N Municipal Wastewater Treated by a SPNDPR System with Different Aeration and Aerobic Times].

An anaerobic (180 min)/aerobic operated sequencing batch reactor (SBR) fed with urban sewage was optimized by regulating the aeration quantity to investigate the deep-level nitrogen (N) and phosphorus (P) removal. The amount of aeration was regulated by adjusting the volume of gas per unit volume of reactor passed in unit time, when the unit is L·(min·L)-1, from 0.125 L·(min·L)-1 gradually to 0.025 L·(min·L)-1, and aerobic times from 3 h to 6 h. The experimental results show that the effluent NH4+-N, NO2--N, NO3--N, and PO43--P concentrations of the optimized SPNDPR system were 0, 8.62, 0.06, and 0.03 mg·L-1. The effluent TN concentration was about 9.22 mg·L-1, and the TN removal efficiency was up to 87.08%. When the aeration quantity was decreased from 0.125 L·(min·L)-1 to 0.100 L·(min·L)-1; then decreased to 0.075 L·(min·L)-1, the nitrification rate of the system recovered and stabilized at 0.16 mg·(L·min)-1. However, when the aeration quantity continuously decreased to 0.050 L·(min·L)-1 and then to 0.025 L·(min·L)-1, the nitrification rate decreased to 0.09 mg·(L·min)-1 and 0.06 mg·(L·min)-1. With reduction of the aeration quantity[from 0.125 L·(min·L)-1 to 0.100, 0.075, 0.050 and 0.025 L·(min·L)-1] and extension of aerobic time (from 3 h to 6 h), the TN removal efficiency increased gradually from 62.82% to 87.08%, and the SND efficiency increased from 19.57% to 72.11%. It was proven that reducing the aeration quantity can enhance the SPND function and deep denitrification by the system was realized. By enhancing the anaerobic intracellular carbon storage and aerobic phosphorus uptake, denitrifying phosphorus removal, partial nitrification, and endogenous nitrification were achieved. The SPNDPR system, after reducing aeration and prolonging aerobic time, was able to realize deep-level denitrification and dephosphorization using low C/N urban sewage.

[Effect of the Influent C/P Ratio on the Nutrient Removal Characteristics of the SNEDPR System].

This study focuses on the nitrogen (N) and phosphorus (P) removal characteristics in a simultaneous nitrification-endogenous denitrification and phosphorus removal (SNEDPR) system at different influent C/P ratios. An extended anaerobic/low aerobic (dissolved oxygen:0.5-1.0 mg·L-1) sequencing batch reactor (SBR) fed with municipal sewage was studied by adjusting different C/P ratios (10, 15, 20, 30, and 60). The experimental results show that the proper reduction of the influent C/P ratio (C/P ratio reduced from 60 to 30) enhances the competitive advantages of phosphorus-accumulating organisms (PAOs) in the SNEDPR system. The highest phosphorus removal efficiency was achieved at a C/P ratio of 30, with the anaerobic phosphorus release rate (PRR) and aerobic phosphorus uptake rate (PUR, used as P/MLSS) reaching 3.5 mg·(g·h)-1 and 4.2 mg·(g·h)-1 respectively, and an average effluent PO43--P concentration below 0.3 mg·L-1. The percentage of PAOs contributing to the storage of endogenesis carbon (PPAO, An) reached 88.1%. However, a poor phosphorus removal performance was observed with further reduction of the influent C/P ratios to 10; both the PO43--P removal efficiency and PPAO, An decreased from 38.1% and 82.4% to 3.1% and 5.3%, respectively. The PRR and PUR were 0.2 mg·(g·h)-1 and 0.24 mg·(g·h)-1, respectively. The COD removal performance was not affected by the decreasing influent C/P ratios; the average COD removal efficiency stabilized at 85%. In addition, the nitrification performance became worse with decreasing C/P ratios (from 60 to 20) because the effluent NH4+-N and NO2--N concentrations increased from 0 and 6.9 mg·L-1 to 5.1 mg·L-1 and 16.2 mg·L-1, respectively. The nitrificaton performance recovered when the C/P ratios further decreased to 10, but the nitrite accumulation was disturbed as both the effluent NH4+-N and NO2--N concentrations reduced to 0. The effluent NO3--N concentration increased from 0.08 mg·L-1 to 14.1 mg·L-1. The SNED efficiency first decreased from 62.1% to 36.4% and then increased to 56.4%. The advantageous competition of glycogen accumulating organisms (GAOs) improved when the influent C/P ratio was lower than 15. The enhancement of the endogenous denitrification ability of GAOs might explain the recovery denitrification performance of the system when the influent C/P ratios decreased from 20 to 10.

[Effect of Different Sludge Retention Time (SRT) Operations on the Nutrient Removal Characteristics of a SNEDPR System].

This study focuses on the nitrogen (N) and phosphorus (P) removal characteristics in a simultaneous nitrification endogenous denitrification and phosphorus removal (SNEDPR) system operating at different sludge retention time (SRT). Four extended anaerobic/low aerobic (dissolved oxygen:0.5-1.0 mg·L-1)-operated sequencing batch reactors (SBRs) fed with municipal sewage were studied at different SRT of 5, 10, 15, and 25 d. The experimental results show that a shorter SRT at an SRT ≥ 10 d enhances the competitive advantage of PAOs in the system and an efficient phosphorus removal performance of the SNEDPR system was achieved at a SRT of 10 d and 15 d. Especially at an SRT of 10 d; the average PPAOs, An was 68.4%, the PRA and PUA reached 31.9 and 34.3 mg·L-1, respectively. The nitrification performance of the system was not affected by SRT changes. The most efficient nitrogen removal performance was achieved at a SRT of 15 d, with a high average TN removal and SNED efficiencies reaching 89.6% and 71.8%, respectively. At a SRT ≥ 10 d, the COD removal performance of the SNEDPR system was also not affected by SRT changes. The COD removal efficiencies were higher than 78%. However, when the SRT was shortened to 5 d, the C, N, and P performances of the system worsened due to the loss of biomass; the SNED and PO43--P removal efficiencies were as low as 5.7% and 0.5%, respectively. In addition, at an SRT=15 d, the sludge-settling performance of the system was the best. The SV and SVI were 20% and 64 mL·g-1, respectively, and the sludge concentration increased with the extension of the SRT. Under long SRT (25 d) operation, the system showed a good resistance to shock loads, but the sedimentation performance of the sludge deteriorated.

[Effect of Influent C/N Ratio on the Nutrient Removal Characteristics of SNEDPR Systems].

To determine the performance of nitrogen and phosphorus removal within a simultaneous nitrification endogenous denitrification system (SNEDPR), an extended anaerobic/low aerobic (dissolved oxygen:0.5-2.0 mg·L-1)-operated sequencing batch reactor (SBR) was fed with simulation wastewater. The SBR was initiated under a constant influent C/N ratio of 10, with the simultaneous enrichment of polyphosphate-accumulating organisms (PAOs). It was then investigated at different influent C/N ratios of 10, 7.5, 5, and 2.5. The experimental results indicated that, when the influent C/N ratio was 10, SNEDPR could be successfully started up. The effluent PO43--P and total nitrogen (TN) concentrations were 0.1 mg·L-1 and 8.1 mg·L-1. PO43--P efficiency, TN efficiency, and SNED efficiency were 99.79%, 89.38%, and 58.0%, respectively. When the influent C/N ratio increased from 5 to 10, the nitrogen and phosphorus removal performance of the system improved with PRA, and SNED efficiency increased from 16.0 m·L-1 and 48.0% to 24.4 mg·L-1 and 69.2%, respectively. When the C/N ratio was 10, the TN and PO43--P removal efficiencies increased to 94.5% and 100%, respectfully. When the C/N ratio was decreased to 2.5, the nitrogen and phosphorus removal performance of the system decreased. The PRA and SNED efficiencies were only 1.36 mg·L-1 and 10%, respectively. During the stable phase of the system (C/N ratio were 10, 7.5 and 5), SNED efficiency reached to 85.9%, with the average effluent concentration of NH4+-N, x--N, and PO43--P being 0.0, 8.1, and 0.1 mg·L-1, respectively.

[Operating Characteristics of a DPR-SNED System Treating Low C/N Municipal Wastewater and Nitrate-containing Sewage].

In order to realize the simultaneous treatment of low C/N municipal wastewater and high nitrate wastewater, a sequencing batch reactor (SBR), inoculated with activated sludge, was used to initiate the denitrifying phosphorus removal coupled with simultaneous nitrification and endogenous denitrification (DPR-SNED). The anaerobic/anoxic/hypoxic durations and dissolved oxygen (DO) concentration were appropriately controlled, and the nitrogen and phosphorus removal characteristics were examined. The experimental results demonstrated that, in the anaerobic/hypoxia operation mode, with an anaerobic duration of 3 h and DO concentration of 0.5-1.0 mg·L-1, the simultaneous nitrification of phosphorus removal (SNEDPR) system successfully began in 60 d. The effluent PO43--P concentration was below 0.5 mg·L-1, the nutrient and COD removal efficiencies were stably maintained above 90% and 80%, respectively, and the SNED efficiency and CODins efficiency reached 70% and 95%, respectively. When the operation mode was anaerobic/anoxic/hypoxic and nitrate-containing sewage was added at the beginning of the anoxic stage, DPR-SNED was achieved with the effluent PO43--P concentration<0.5 mg·L-1, nutrient and COD removal efficiencies above 88% and 90%, respectively, and SNED efficiency and CODins efficiency maintained at 62% and 90%, respectively. After the successful initiation of DPR-SNED, enhanced intracellular carbons storage was achieved by phosphorus-and glycogen-accumulating organisms using the limited carbons in raw municipal wastewater to provide sufficient carbon sources for subsequent nutrient removal. In addition, the endogenous partial denitrification ensured the efficient nitrogen removal performance of the DPR-SNED system at low C/N conditions (average 4).

[Simultaneous Nitrogen and Phosphorus Removal Characteristics of An Anaerobic/Aerobic Operated SPNDPR System Treating Low C/N Urban Sewage].

This study focused on the nitrogen (N) and phosphorus (P) removal performance optimization of simultaneous partial nitrification-endogenous denitrification and phosphorus removal (SPNDPR) systems. An anaerobic (180 min)/aerobic operated sequencing batch reactor (SBR) fed with domestic wastewater was used for investigating the startup and optimization of SPNDPR by regulating the aeration rate and aerobic duration time. The experimental results showed that at an aerobic aeration rate of 0.8 L·min-1 and aerobic duration time of 150 min, the effluent PO43--P concentration was about 1.5 mg·L-1, with the effluent NH4+-N and NO3--N concentrations gradually decreasing from 10.28 and 8.14 mg·L-1 to 0 and 2.27 mg·L-1, respectively, and effluent NO2--N concentration increasing to 1.81 mg·L-1. When the aeration rate was increased to 1.0 L·min-1 and the aerobic duration time was shortened to 120 min, the phosphorus removal and partial nitrification-endogenous performance of the system gradually increased, but the total nitrogen (TN) removal performance initially decreased and then gradually increased. The final effluent PO43--P and NH4+-N were stably below 0.5 and 1.0 mg·L-1, respectively, aerobic nitrite accumulation and simultaneous nitrification-endogenous denitrification (SND) efficiencies were 98.65 and 44.20%, respectively, and TN removal efficiency was 79.78%. The concurrence of aerobic phosphorus absorption, denitrifying phosphorus removal, partial nitrification, and nitrification-endogenous in the aerobic stage of the SPNDPR system ensured the simultaneous removal of N and P from low C/N wastewater.