Simultaneous Nitrification-endogenous Denitrification Research Articles

Simultaneous Nitrification–Endogenous Denitrification with Phosphorus Removal Sustained by Aeration-Free Filamentous Microalgal–Bacterial Granular Sludge for Low C/N Wastewater Treatment

Simultaneous Nitrification–Endogenous Denitrification with Phosphorus Removal Sustained by Aeration-Free Filamentous Microalgal–Bacterial Granular Sludge for Low C/N Wastewater Treatment

Combined Phototrophic Simultaneous Nitrification-Endogenous Denitrification with Phosphorus Removal (P-SNDPR) System Treating Low Carbon to Nitrogen Ratio Wastewater for Potential Carbon Neutrality.

Conventional biological nutrient removal processes rely on external aeration and produce significant carbon dioxide (CO2) emissions. This study constructed a phototrophic simultaneous nitrification-denitrification phosphorus removal (P-SNDPR) system to treat low carbon to nitrogen (C/N) ratios wastewater and investigated the impact of sludge retention time (SRT) on nutrient removal performance, nitrogen conversion pathway, and microbial structure. Results showed that the P-SNDPR system at SRT of 15 days had the highest nutrient removal capacity, achieving over 85% and 98% removal of nitrogen and phosphorus, respectively, meanwhile maintaining minimal CO2 emissions. Nitrogen removal was mainly through assimilation at SRTs of 5 and 10 days, and nitrification-denitrification at SRTs of 15 and 20 days. Stable partial nitrification was facilitated by photoinhibition and low DO levels. Flow cytometry sorting technique results revealed SRT drove community structural changes in translational activity (BONCAT+) microbes, where BONCAT+ microbes were mainly simultaneous nitrogen and phosphorus removal bacteria (Candidatus Accumulibacter), denitrifying bacteria (Candidatus Competibacter and Plasticicumulans), ammonia-oxidizing bacteria (Nitrosomonas), and microalgae (Chlorella and Dictyosphaerium). The P-SNDPR system represents a novel, carbon-neutral process for efficient nutrient removal from low C/N ratio wastewater without aeration and external carbon source additions.

Exploring organic matter conversion pathway and its effect on nitrogen removal in tidal flow constructed wetlands

Achieving effective nitrogen removal remains a significant challenge faced by constructed wetlands. Although organic matter is a crucial factor influencing nitrogen removal, little attention has been paid to the impact of organic matter conversion pathways on nitrogen removal in constructed wetlands. Here, we showed that endogenous microorganisms performing carbon internalization could be easily enriched in tidal flow constructed wetlands (TFCWs) under its special rhythmic cycle of anaerobic/aerobic operational mode. Endogenous microorganisms could translate influent carbon sources into intracellular carbons during the anaerobic stage and supply the carbon source for endogenous denitrification after the aerobic stage (rest period). Based on these findings, an innovative combined TFCW and Nitrifying-CW system was developed, and robust total nitrogen (TN) removal (82% on average) was achieved even under carbon source limiting conditions. This performance was a substantial improvement compared to the conventional single bed TFCW with multiple “tides” (corresponding to the multiple contact/rest periods) with TN removal of only 54% on average. Simultaneous nitrification-endogenous denitrification (SNED) was found to be the major nitrogen removal pathway in the proposed system. Compared with classical nitrification-denitrification, simultaneous nitrification-endogenous denitrification brings high nitrogen conversion rates and significantly reduces the demand for oxygen and organic carbon. Furthermore, microbial community analysis indicated that endogenous microorganisms such as Candidatus_Competibacter and Defluviicoccus were successfully enriched, accounting for 50.73% and 3.46% in CW1, and 25.25% and 1.76% in CW2, respectively. Together, these mechanisms allow the proposed system to achieve efficient TN removal.

The successful application of light to the system of simultaneous nitrification/endogenous denitrification and phosphorus removal: Promotion of partial nitrification and glycogen accumulation metabolism

Partial nitrification (PN) and high glycogen accumulating metabolism (GAM) activity are the basis for efficient nitrogen (N) and phosphorus (P) removal in simultaneous nitrification endogenous denitrification and phosphorus removal (SNDPR) systems. However, achieving these processes in practical operations is challenging. This study proposes that light irradiation is a novel strategy to enhance the nutrient removal performance of the SNDPR system with low carbon to nitrogen ratios (C/N of 3.3–4.1) domestic wastewater. Light energy densities (Es) of 55–135 J/g VSS were found to promote the activity of ammonia-oxidizing bacteria (AOB) and GAM, while inhibiting the activity of nitrite-oxidizing bacteria (NOB) and polyphosphate accumulating metabolism (PAM). Long-term exposure to different light patterns at Es of 55–135 J/g VSS revealed that continuous light rapidly achieved PN by inhibiting NOB activity and promoted the growth of glycogen accumulating organisms (GAOs), allowing the removal of above 82 % N and below 80 % P. Intermittent light maintained stable PN by inhibiting the activity and growth of NOB and promoted the growth of polyphosphate accumulating organisms (PAOs) with high GAM activity (Accmulibacer IIC-ii and IIC-iii), allowing the removal of above 82 % N and 95 % P. Flow cytometry and enzyme activity assays showed that light promoted GAM-related enzyme activity and the metabolic activity of partial Accmulibacer II over other endogenous denitrifying bacteria, while inhibiting NOB translation activity. These findings provide a new approach for enhancing nutrient removal, especially for achieving PN and promoting GAM activity, in SNDPR systems treating low C/N ratio domestic wastewater using light irradiation.

Start-up mechanism of simultaneous nitrification-endogenous denitrification process for treatment of low C/N wastewater: Insights into reactor performance and microbial community dynamics

The aim of this study is to propose a start-up strategy by reducing DO concentration, optimizing the duration of the operation mode and gradually decreasing influent C/N for simultaneous nitrification-endogenous denitrification (SNED) process to economically and effectively treat low carbon/nitrogen (C/N) wastewater, and to explore the start-up mechanism from the microbiome. After start-up, the efficiency of simultaneous nitrification-denitrification, nitrogen loading rate, and nitrogen removing rate increased from 8.3%, 60 mg N/L⋅d, and 55.2 mg N/L⋅d to 60.9%, 160 mg N/L⋅d, and 143.6 mg N/L⋅d, respectively. Meanwhile, the total nitrogen removal efficiency and the conversion of intracellular carbon were maintained at 90.2% and 89.1%. Correspondingly, microbial community diversity was significantly inhibited, however, the key functional microbes remarkably increased such as Defluviicoccus (denitrifying glycogen accumulating organisms), Paracoccus (heterotrophic nitrification and aerobic denitrification bacteria), etc. The reduction of heterotrophs led to the streamlining of network scale and complexity. But, the emergence of more positive interaction in functional subnetworks, stable module numbers, and the redundancy of keystone taxa contributes to the targeted regulation of functional microorganisms, as further confirmed by increased community function. Deterministic process dominated the community assembly during start-up period, where homogenizing selection is the main driving force for key functional microbes, e.g., Defluviicoccus and Paracoccus. This study provided key information for practical application of SNED process and laid the foundation for cost-effective treatment of low C/N wastewater.

Strengthening anoxic glycogen consumption in SNEDPR-CW as a strategy to control PAO–GAO competition under carbon limited condition

Cooperation between Phosphate and Glycogen Accumulating Organisms (PAOs and GAOs) plays a pivotal role in nutrients removal in simultaneous nitrification endogenous denitrification and phosphorous removal (SNEDPR) systems. Recent findings have expanded the application of SNEDPR from activated sludge system to constructed wetland (CW). However, how to regulate competition between PAOs and GAOs in SNEDPR-based CW is still unclear. Here we showed that, GAOs could easily gain dominance over PAOs in SNEDPR-CW under alternating anaerobic/aerobic (A/O) operational mode. Shortening aerobic hydraulic retention time (HRT) at low oxygen concentration was benefit for simultaneous nitrification endogenous denitrification (SNED) and denitrifying dephosphatation but would reduce the overall phosphorus uptake rate and lead to high phosphorus effluent concentrations. Extended aerobic HRT promoted the proliferation of aerobic GAOs over PAOs, decreasing both enhanced biological phosphorus removal (EBPR) and SNED performance. Surprisingly, by switching the operation of system to alternating anaerobic/aerobic/anoxic (A/O/A) mode, an extraordinary nutrients removal performance with mean nitrogen and phosphorus removal efficiency of 84.57% and 89.37% was achieved under carbon sources limited condition. Stoichiometric analysis demonstrated that adding anoxic stage strengthened the intracellular glycogen oxidization of GAOs for denitrification which compromised its subsequent anaerobic carbon sources uptake and PHA storage and provided sufficient carbon sources for PAOs. Microbial community analysis showed that numerical ratio of GAOs to PAOs decreased from 6.67 under A/O to 4.89 under A/O/A mode, which further indicated strengthening glycogen denitrification of GAOs should be an effective way to regulate microbial competition in order to obtain a desired nutrients removal performance in SNEDPR-CW.

Metagenomic insights into the explanation of biofilter performance distinction induced by dissolved oxygen increment

In this study, a set of novel double-layer-packed sequencing batch biofilm reactors (SBBRs) were constructed to treat secondary effluent with a low carbon-to-nitrogen ratio (C/N = 3.0). All the reactors were operated under identical conditions except dissolved oxygen (DO) in the aerobic section, which was controlled at seven different levels (0.5, 1.0, 1.5, 2.0, 2.5, 3.5 and 4.5 mg/L). It was found that the aerobic section contributed most to performance at an optimal DO of 2.0 mg/L. The change in DO accounted for the varied bacterial communities, which resulted in different performances. Both bacterial community structure and potential functions displayed close associations with performance implying nutrients were mainly removed by simultaneous nitrification-endogenous denitrification and phosphorus removal (SNEDPR) process. Moreover, the proliferation of nonfunctional nitrifiers/denitrifiers led to the inhibition of denitrification resulting in a lower removal of total dissolved nitrogen and phosphorus.

Synergistic simultaneous nitrification-endogenous denitrification and EBPR for advanced nitrogen and phosphorus removal in constructed wetlands

• Achieving EBPR with high phosphorus removal performance in CWs. • SNED was dominant nitrogen removal pathway in EBPR-CW without specific control. • Coordination of nitrifiers, APAOs, DPAOs and DGAOs facilitated N and P removal. • Average removal efficiency of PO 4 3 - -P and TN were 92.68% and 96.21%, respectively. This study assessed the feasibility of achieving phosphorus removal through enhanced biological phosphorus removal (EBPR) in an intermittent aeration constructed wetland (CW). The results showed that within just 3 weeks, a polyphosphate-accumulating organisms (PAOs)-enriched biofilm was successfully established in the CW by integrating a pre-anaerobic stage, intermittent aeration and periodic PO 4 3- -P release strategies. This strategy enabled a high PO 4 3- -P removal efficiency of 92.68% using normal gravel as a substrate. Moreover, simultaneous nitrification-endogenous denitrification (SNED) was found to be the main nitrogen removal pathway in the EBPR-CW without specific control. SNED significantly reduces the demand for oxygen and organic carbon compared with classical nitrification-denitrification, and allowed the system to reach 96.21% total nitrogen (TN) removal even under carbon limiting conditions. With control experiment, EBPR combined with SNED in CWs improved the phosphorus and nitrogen removal by about 73% and 48%, respectively, compared with normal nitrification-denitrification and media storage-based nitrogen and phosphorus removal processes. Based on stoichiometric analysis, 71.25% of PO 4 3- -P was removed by aerobic PAOs (APAOs), with 27.50% removed by DPAO Ni (denitrifying PAOs using nitrite as electron acceptor) and 1.25% removed by DPAO Na (denitrifying PAOs using nitrate as electron acceptor). DGAOs were the main organisms providing nitrite to DPAOs. Overall, our results demonstrate a novel approach to combine SNED with EBPR for advanced nitrogen and phosphorus removal in CWs.

Evaluation of functional microbes in explaining biofilter performance during start-up1

As an effective alternative for dissolved nitrogen removal, biofilter normally associates its treatment performance to various microbial communities that are shaped by different structural/operational conditions. In this study, a set of four different biofilters including MAVF (mature aerated vertical flow), NAVF (new aerated vertical flow), NVF (new non-aerated vertical flow) and BHF (baffled non-aerated horizontal flow) were employed to treat simulated domestic wastewater and the potential relationships between purification performance and functional microbes during start-up were explored. It was found that the four filters could be well clustered or separated according to their performance with the MAVF having the highest nitrogen removal, i.e. 52% for total nitrogen (TN) and 85% for total inorganic nitrogen (TIN). Moreover, TIN should be mainly removed by simultaneous nitrification-endogenous denitrification in aerobic/facultative systems. Microbial community analysis revealed that the entire and the first 20 dominant taxa, nitrifiers, and methanotrophs all displayed a similar clustering pattern, which was consistent with that of treatment performance indicating the defined purification was not only determined by functional microbes but also related to the entire microbial community. Nevertheless, Planctomycetes, Candidatus Saccharibacteria, and Candidatus Accumulibacter all displayed a weak association with performance implying these functional microbes contributed little to performance. This was probably attributed to the low inflow load as well as the lack of alternate anaerobic/aerobic conditions inside the filter bed. These findings might be helpful for biofilter optimization or management.

Effect of Step Aeration on a Municipal Sewage Aerobic Granular Sludge System

In order to achieve simultaneous nitrogen and phosphorus removal in low-C/N urban sewage, a sequencing batch reactor (SBR) was operated in anaerobic/aerobic (A/O) mode. Keeping the total aeration volume at 900 L, the aeration strategy was adjusted. The uniform aeration of 2.81 L·(h·L)-1 was changed to "high/low aeration" with high strength 4.22 L·(h·L)-1before low strength 1.88 L·(h·L)-1, and "low/high aeration" with low strength 1.88 L·(h·L)-1 before high strength 4.22 L·(h·L)-1. The experiment investigated the nitrogen and phosphorus removal performance and sludge characteristics of the system under different aeration strategies. The results showed that the nitrogen and phosphorus removal performances of the system under high/low aeration were the best. The effluent concentrations of NH4+-N, NO2--N, NO3--N, and TP were 0, 0.15, 8.12, and 0.04 mg·L-1, respectively. The removal rates of total nitrogen (TN) and total phosphorus (TP) were as high as 78.33% and 99.19%, respectively. Simultaneous nitrification endogenous denitrification (SNED) was clear, with the SNED ratio at 77.08%. Compared with uniform aeration, the system nitrification rate and denitrification rate increased, and the denitrification rate reached 14.33 mg·(g·h)-1, which was the maximum value during the whole operation; the solidity, sedimentation performance, and stability of granular sludge were improved, and the sludge volume index (SVI) was 23.49 mL·g-1. After adjusting the aeration strategy to low/high aeration, the nitrogen and phosphorus removal performance of the system deteriorated, and the removal rates of TN and TP were reduced to 51.26% and 58.32%, respectively. However, the system had the best nitrification performance with ammonia oxidation rate and nitrate production rate at 14.92 mg·(g·h)-1and 7.50 mg·(g·h)-1, respectively, which were their maximum values during the whole operation. Simultaneously, the filamentous bacteria in the granular sludge multiplied, the granular structure became loose, the sedimentation and stability all worsened, and the SVI rose to 40.76 mL·g-1.

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 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.

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.

Combining simultaneous nitrification-endogenous denitrification and phosphorus removal with post-denitrification for low carbon/nitrogen wastewater treatment

Due to the limited nutrient removal from low carbon/nitrogen (⩽4) wastewater, a process combined simultaneous nitrification-endogenous denitrification and phosphorus removal (SNDPR) with post-denitrification (PD) in a SBR was proposed for deep-level nutrient removal without external carbon addition. SNDPR driven by PAOs and GAOs reduced PO43−-P (98.3%) and partial TN (59.0%) at low DO conditions (0.5±0.1mg/L), and post-dentrification achieved further NOX− (produced by SNDPR) removal (24.0%) anoxically by utilizing the residual intracellular polymers in GAOs. Combined control of anaerobic/aerobic/anoxic durations and low DO inhibition to aerobic GAOs and NOB conducted partial nitrification-endogenous denitrification (PNED) (66%), which saved 44.3% intracellular polymers to further reduce 64% TN in effluent. After 115-day operation, the average effluent PO43−-P and TN concentrations were 0.4 and 3.9mg/L, respectively, with 92.1% of TN removal. Highly enriched PAOs (36%±2%), GAOs (22%±2%) and AOB (15%±3%) over NOB (3%±1%) facilitated P uptake, PNED and post-denitrification in the SNDPR-PD system.

A novel stoichiometries methodology to quantify functional microorganisms in simultaneous (partial) nitrification-endogenous denitrification and phosphorus removal (SNEDPR)

Although efficient removal of carbon (C), nitrogen (N) and phosphorous (P) from wastewater with low C/N ratio was achieved in anaerobic/aerobic simultaneous nitrification-endogenous denitrification and phosphorus removal (SNEDPR) systems, the removal pathways and metabolic transformations in this complex system are unclear. This work targeted at developing the stoichiometric models for denitrifying glycogen organisms (DGAOs) via nitrite and nitrate (DGAONi and DGAONa), and demonstrating a novel methodology to quantify diverse functional microorganisms (e.g. ammonia and nitrite oxidizing bacteria, aerobic phosphorus accumulating organisms (APAOs), denitrifying PAOs (DPAOs) and aerobic GAOs (AGAOs)) for the removal of C, N and P. The results showed that the anaerobic intracellular carbon storage (CODintra) was mainly accomplished by GAOs, and PAOs were only responsible for about 40% of CODintra through a stable P release. At the aerobic stage, 84.9% of P was removed by APAOs with 15.1% left by DPAOs, while 64.6% of N was removed by DGAOs (45.8% by DGAONi and 18.8% by DGAONa) with 18.1% by DPAOs and 17.3% by bacterial growth. High proportion of N removal via nitrite (partial nitrification-endogenous denitrification) (71%) saved 7.3% aeration and 38% intracellular carbon demand. However, AGAOs still activated well at the aerobic intercellular carbon consumption, which limited the further improvement of N removal efficiency. By elucidating the nutrient removal pathways among diverse functional microorganisms, the methodology developed in this study could accelerate the nutrient removal in the SNEDPR process.

Treating low carbon/nitrogen (C/N) wastewater in simultaneous nitrification-endogenous denitrification and phosphorous removal (SNDPR) systems by strengthening anaerobic intracellular carbon storage

A novel simultaneous nitrification denitrification and phosphorous removal-sequencing batch reactor (SNDPR-SBR) enriched with PAOs (phosphorus accumulating organisms), DPAOs (denitrifying PAOs), and GAOs (glycogen accumulating organisms) at the ratio of 2:1:1 was developed to achieve the simultaneous nutrient and carbon removal treating domestic wastewater with low carbon/nitrogen ratio (≤3.5). The SNDPR system was operated for 120 days at extended anaerobic stage (3 h) and short aerobic stage at low oxygen concentration (2.5 h) with short sludge retention time (SRT) of 10.9 d and hydraulic retention time (HRT) of 14.6 h. The results showed that at the stable operating stage, the average effluent chemical oxygen demand (COD) and PO43−–P concentrations were 47.2 and 0.2 mg L−1, respectively, the total nitrogen (TN) removal efficiency was 77.7%, and the SND efficiency reached 49.3%. Extended anaerobic stage strengthened the intracellular carbon (mainly poly-β-hydroxybutyrate, PHB) storage, efficiently utilized the organic substances in wastewater, and provided sufficient carbon sources for denitrification and phosphorus uptake without external carbon addition. Short aerobic stage at low oxygen concentration (dissolved oxygen (DO): 1 ± 0.3 mg L−1) achieved a concurrence of nitrification, endogenous denitrification, denitrifying and aerobic phosphorus uptake, and saved about 65% energy consumption for aeration. Microbial community analysis demonstrated that P removal was mainly performed by aerobic PAOs while N removal was mainly carried out by denitrifying GAOs (DGAOs), even though DPAOs were also participated in both N and P removal.