Temperature-resilient superior performances by coupling partial nitritation/anammox and iron-based denitrification with granular formation.

Partial nitritation-anammox (PN/A), an energy-neutral process, is widely employed in the treatment of nitrogen-rich wastewater. However, the intrinsic nitrate accumulation limits the total nitrogen (TN) removal, and the practical application of PN/A continues to face a significant challenge at low temperatures (<15 °C). Here, an integrated partial nitritation-anammox and iron-based denitrification (PNAID) system was developed to address the concern. Two up-flow bioreactors were set up and operated for 400 days, with one as the control group and the other as the experiment group with the addition of Fe0. In comparison to the control group, the experiment group with the Fe0 supplement showed better nitrogen removal during the entire course of the experiment at different temperature levels. Specifically, the TN removal efficiency of the control group decreased from 82.9 % to 53.9 % when the temperature decreased from 30 to 12 °C, while in stark contrast, the experiment group consistently achieved 80 % of TN removal in the same condition. Apart from the enhanced nitrogen removal, the experiment group also exhibited better phosphorus removal (10.6 % versus 74.1 %) and organics removal (49.5 % versus 65.1 %). The enhanced and resilient nutrient removal performance of the proposed integrated process under low temperatures appeared to be attributed to the compact structure of granules and the increased microbial metabolism with Fe0 supplement, elucidated by a comprehensive analysis including microbial-specific activity, apparent activation energy, characteristics of granular sludge, and metagenomic sequencing. These results clearly confirmed that Fe0 supplement not only improved nitrogen removal of PN/A process, but also conferred a certain degree of robustness to the system in the face of temperature fluctuations.

Iron-electrolysis assisted anammox/denitrification system for intensified nitrate removal and phosphorus recovery in low-strength wastewater treatment.

Two iron-electrolysis assisted anammox/denitrification (EAD) systems, including the suspended sludge reactor (ESR) and biofilm reactor (EMR) were constructed for mainstream wastewater treatment, achieving 84.51±4.38 % and 87.23±3.31 % of TN removal efficiencies, respectively. Sludge extracellular polymeric substances (EPS) analysis, cell apoptosis detection and microbial analysis demonstrated that the strengthened cell lysate/apoptosis and EPS production acted as supplemental carbon sources to provide new ecological niches for heterotrophic bacteria. Therefore, NO3--N accumulated intrinsically during anammox reaction was reduced. The rising cell lysis and apoptosis in the ESR induced the decline of anammox and enzyme activities. In contrast, this inhibition was scavenged in EMR because of the more favorable environment and the significant increase in EPS. Moreover, ESR and EMR achieved efficient phosphorus removal (96.98±5.24 % and 96.98±4.35 %) due to the continued release of Fe2+ by the in-situ corrosion of iron anodes. The X-ray diffraction (XRD) indicated that vivianite was the dominant P recovery product in EAD systems. The anaerobic microenvironment and the abundant EPS in the biofilm system showed essential benefits in the mineralization of vivianite.

Partial denitrifying phosphorus removal coupling with anammox (PDPRA) enables synergistic removal of C, N, and P nutrients from municipal wastewater: A year-round pilot-scale evaluation.

Applying anaerobic ammonium oxidation (anammox) in municipal wastewater treatment plants (MWWTPs) can unlock significant energy and resource savings. However, its practical implementation encounters significant challenges, particularly due to its limited compatibility with carbon and phosphorus removal processes. This study established a pilot-scale plant featuring a modified anaerobic-anoxic-oxic (A2O) process and operated continuously for 385 days, treating municipal wastewater of 50 m3/d. For the first time, we propose a novel concept of partial denitrifying phosphorus removal coupling with anammox (PDPRA), leveraging denitrifying phosphorus-accumulating organisms (DPAOs) as NO2- suppliers for anammox. 15N stable isotope tracing revealed that the PDPRA enabled an anammox reaction rate of 6.14 ± 0.18 µmol-N/(L·h), contributing 57.4 % to total inorganic nitrogen (TIN) removal. Metagenomic sequencing and 16S rRNA amplicon sequencing unveiled the co-existence and co-prosperity of anammox bacteria and DPAOs, with Candidatus Brocadia being highly enriched in the anoxic biofilms at a relative abundance of 2.46 ± 0.52 %. Finally, the PDPRA facilitated the synergistic conversion and removal of carbon, nitrogen, and phosphorus nutrients, achieving remarkable removal efficiencies of chemical oxygen demand (COD, 83.5 ± 5.3 %), NH4+ (99.8 ± 0.7 %), TIN (77.1 ± 3.6 %), and PO43- (99.3 ± 1.6 %), even under challenging operational conditions such as low temperature of 11.7 °C. The PDPRA offers a promising solution for reconciling the mainstream anammox and the carbon and phosphorus removal, shedding fresh light on the paradigm shift of MWWTPs in the near future.

Simultaneous nitrogen and phosphorus removal through an integrated partial-denitrification/anammox process in a single UAFB system.

With the aim of obtaining enhanced nitrogen removal and phosphate recovery in mainstream sewage, we examined an integrated partial-denitrification/anaerobic ammonia oxidation (PD/A) process over a period of 189 days to accomplish this goal. An up-flow anaerobic fixed-bed reactor (UAFB) used in the integrated PD/A process was started up with anammox sludge inoculated and the influent composition controlled. Results showed that the system achieved a phosphorus removal efficiency of 82% when the influent concentration reached 12.0 mg/L. Batch tests demonstrated that stable and efficient removal of chemical oxygen demand (COD), nitrogen, and phosphorus was achieved at a COD/NO3--N ratio of 3.5. Scanning electron microscope (SEM) and X-ray diffraction (XRD) analysis indicated that hydroxyapatite was the main crystal in the biofilm. Furthermore, substrate variation along the axial length of UAFB indicated that partial denitrification and anammox primarily took place near the reactor's bottom. According to a microbiological examination, 0.4% of the PD/A process's microorganisms were anaerobic ammonia oxidizing bacteria (AnAOB). Ca. Brocadia, Ca. Kuenenia, and Ca. Jettenia served as the principal AnAOB generals in the system. Thauera, Candidatus Accumulibacter, Pseudomonas, and Acinetobacter, which together accounted for 27% of the denitrifying and phosphorus-accumulating bacteria, were helpful in advanced nutrient removal. Therefore, the combined PD/A process can be a different option in the future for sewage treatment to achieve contemporaneous nutrient removal.

Metagenomics insight into the long-term effect of ferrous ions on the mainstream anammox system.

Anaerobic ammonium oxidation (anammox) bacteria have a high requirement for iron for their growth and metabolism. However, it remains unclear whether iron supplementation can sustain the stability of mainstream anammox systems at varying temperatures. Here, we investigated the long-term effects of Fe2+ on the mainstream anammox systems. Our findings revealed that the nitrogen removal efficiency (NRE) of the anammox system supplemented with 5 mg/L Fe2+ decreased from 76.5 ± 0.76% at 35 °C to 39.0 ± 9.9% at 25 °C. Notably, higher dosages of Fe2+ (15 mg/L and 30 mg/L) inhibited the anammox system, resulting in NREs of 15.9 ± 8.1% and 2.5 ± 1.1% at 25 °C, respectively. The results of microbial communities and function profiles suggested that the high Fe2+ dosage seriously affected the iron assimilation and utilization in the mainstream anammox system. This was evident from the decreased abundance of genes associated with Fe(II) transport and uptake, which in turn hindered the biosynthesis of intracellular iron-cofactors, resulting in decrease in the absolute abundance of Candidatus Brocadia, a key anammox bacterium, as well as a decline in NRE. Furthermore, our results showed that the anammox process was more susceptible to iron supplementation at 25 °C compared to 35 °C, which may be due to the oxidative stress reactions induced by combined lowered temperature and a high Fe2+ dosage. Overall, these findings offer a deeper understanding of the effect of iron in mainstream anammox systems, which can contribute to improved stability maintenance and effectiveness of anammox processes.

Sidestream bio-P and mainstream anammox in a BNR process with upstream carbon capture.

The integration of biological phosphorus removal (bio-P) and shortcut nitrogen removal (SNR) processes is challenging because of the conflicting demands on influent carbon: SNR allows for upstream carbon diversion, but this reduction of influent carbon (especially volatile fatty acids [VFAs]) prevents or limits bio-P. The objective of this study was to achieve SNR, either via partial nitritation/anammox (PNA) or partial denitrification/anammox (PdNA), simultaneously with biological phosphorus removal in a process with upstream carbon capture. This study took place in a pilot scale A/B process with a sidestream bio-P reactor and tertiary anammox polishing. Despite low influent rbCOD concentrations from the A-stage effluent, bio-P occurred in the B-stage thanks to the addition of A-stage WAS fermentate to the sidestream reactor. Nitrite accumulation occurred in the B-stage via partial denitrification and partial nitritation (NOB out-selection), depending on operational conditions, and was removed along with ammonia by the tertiary anammox MBBR, with the ability to achieve effluent TIN less than 2 mg/L. PRACTITIONER POINTS: A sidestream reactor with sufficient fermentate addition enables biological phosphorus removal in a B-stage system with little-to-no influent VFA. Enhanced biological phosphorus removal is not inhibited by intermittent aeration and is stable at a wide range of process SRTs. Partial nitritation and partial denitrification are viable routes to produce nitrite within an A/B process with sidestream bio-P, for downstream anammox in a polishing MBBR.

Anammox-Mediated Hydroxyapatite Granules: Physicochemical Properties, 3D Hierarchy, and Biofilm Thickness.

Biomineralization inspired the development of simultaneous biological transformations and chemical precipitation for simultaneous nitrogen removal and phosphorus recovery from wastewater, which could compensate for the incapacity of phosphorus management in the new biological route of anaerobic ammonium oxidation (anammox). In this study, we strengthened anammox-mediated biomineralization by long-term feeding of concentrated N, P, and Ca substrates, and a self-assembled matrix of anammox bacteria and hydroxyapatite (HAP) was fabricated in a granular shape, defined as HAP-anammox granules. HAP was identified as the dominant mineral using elemental analysis, X-ray diffraction, and Raman spectroscopy. The intensive precipitation of HAP resulted in a higher inorganic fraction and substantially improved settleability of anammox biomass, which facilitated HAP precipitation by acting as nucleation and metabolically elevated pH. By using X-ray microcomputed tomography, we visually represented the hybrid texture of interwoven HAP pellets and biomass, the core-shell layered architecture of different-sized HAP-anammox granules, and their homogeneously regulated thickness of the outer biofilm (from 118 to 635 µm). This unique architecture endows HAP-anammox granules with outstanding settleability, active biofilm, and tightly bonded biofilm with the carrier, which may explain the excellent performance of these HAP-anammox granules under various challenging operational conditions in previous studies.

Achieving advanced nitrogen removal from low-carbon municipal wastewater using partial-nitrification/anammox and endogenous partial-denitrification/anammox.

To achieve advanced nitrogen removal from low-carbon wastewater, a partial-nitrification/anammox and endogenous partial-denitrification/ anammox (PN/A-EPD/A) process was developed in a sequential batch biofilm reactor (SBBR). Advanced nitrogen was achieved with the effluent total nitrogen (TN) of 3.29 mg/L when the influent COD/TN and the TN were 2.86 and 59.59 mg/L, respectively. This was attributed to a stable PN/A-EPD/A, which was achieved through the integration of four strategies, including treating the inoculated sludge with free nitrous acid, inoculating anammox biofilm, discharging excess activated sludge and residual ammonium at the end of oxic stage. The 16S rRNA high-throughput sequencing results demonstrated that anammox bacteria coexisted with ammonia oxidizing bacteria, nitrite oxidizing bacteria, denitrifying glycogen accumulating organisms (DGAOs) and denitrifying phosphorus accumulating organisms (DPAOs) in biofilms. The abundance of anammox bacteria in the inner layer of the biofilm is higher, while that of DGAOs and DPAOs is higher in the outer layer.

Improved Properties and Enhancement Strategies of Hydroxyapatite-Based Functional Granular Sludge for a High-Rate Partial Nitritation/Anammox System.

Retaining sufficient anammox bacteria (AnAOB) while keeping the anammox-based process stable is the focus of the study of anammox technology, especially in a one-stage partial nitritation/anammox (PNA) process. The use of hydroxyapatite (HAP) granules in an anammox-based process is innovative for its potential to improve the nitrogen removal rate and achieve simultaneous removal of phosphorus. In this study, the HAP-based granular sludge was employed using enhancement strategies for an excellent nitrogen removal performance in a one-stage PNA process. Compared to those of other granular sludge PNA systems, a remarkable sludge volume index of 7.8 mL/g and an extremely high mixed liquor volatile suspended solids of 15 g/L were achieved under a low hydraulic retention time of 2 h. Consequently, an unprecedented nitrogen removal rate as high as 4.8 kg N/m3/d at 25 °C was obtained under a nitrogen loading rate of 6 kg N/m3/d. After a long-term operation of 870 days, the enhancement strategies underlying the superior performance of the granular sludge were identified. These findings clearly demonstrate that the enhancement strategies are crucial for the superior operating performance of the PNA process, and they can promote the application of the anammox-based process.

Towards advanced simultaneous nitrogen removal and phosphorus recovery from digestion effluent based on anammox-hydroxyapatite (HAP) process: Focusing on a solution perspective.

In this paper, the state-of-the-art information on the anammox-HAP process is summarized. The mechanism of this process is systematically expounded, the enhancement of anammox retention by HAP precipitation and the upgrade of phosphorus recovery by anammox process are clarified. However, this process still faces several challenges, especially how to deal with the âˆ¼ 11% nitrogen residues and to purify the recovered HAP. For the first time, an anaerobic fermentation (AF) combined with partial denitrification (PD) and anammox-HAP (AF-PD-Anammox-HAP) process is proposed to overcome the challenges. By AF of the organic impurities of the anammox-HAP granular sludge, organic acid is produced to be used as carbon source for PD to remove the nitrogen residues. Simultaneously, pH of the solution drops, which promotes the dissolution of some inorganic purities such as CaCO3. In this way, not only the inorganic impurities are removed, but the inorganic carbon is supplied for anammox bacteria.

Partial Nitritation/Anammox and biological phosphorus removal integration in a single bioreactor under mainstream conditions.

Anammox-based nitrogen removal and enhanced biological phosphorus removal (EBPR) are increasingly applied for nutrient removal from wastewater, but are typically operated in separate reactors. Here, a novel process for integrated partial nitritation/anammox (PN/A) and EBPR in a single reactor employing integrated fixed film activated sludge was tested. The reactor was fed with mainstream municipal wastewater (5.4 ± 1.3 g COD/g N) at 20 °C for 243 days. Robust ammonium, total inorganic nitrogen, and orthophosphate removal efficiencies of 94 ± 4 %, 87 ± 7 % and 92 ± 7 % were achieved. Nitrite-oxidizing organisms suppression and ammonia-oxidizing organisms retention were achieved via solids retention time control, intermittent aeration, and suspended versus attached biomass population segregation. The contribution of anammox to nitrogen removal increased from 24 % to 74 %. In parallel, a substantial enrichment of Tetrasphaera polyphosphate accumulating organisms was observed. This work demonstrates a novel intensified bioprocess coupling PN/A and EBPR in the same reactor for efficient nutrient removal from wastewater.

Ultra-high nitrogen removal from real municipal wastewater using selective enhancement of glycogen accumulating organisms (GAOs) in a partial nitrification-anammox (PNA) system.

Integrating endogenous denitrification (ED) into partial nitrification-anammox (PNA) systems by adequately utilizing organics in municipal wastewater is a promising approach to improve nitrogen removal efficiency (NRE). In this study, a novel strategy to inhibit phosphorus-accumulating organisms (PAOs) by inducing phosphorus release and exclusion was adopted intermittently, optimizing organics allocation between PAOs and glycogen-accumulating organisms (GAOs). Enhanced ED-synergized anammox was established to treat real municipal wastewater, achieving an NRE of 97.5±2.2% and effluent total inorganic nitrogen (TIN) of less than 2.0 mg/L. With low poly-phosphorus (poly-P) levels (poly-P/VSS below 0.01 (w/w)), glycogen accumulating metabolism (GAM) acquired organics exceeded that of phosphorus accumulating metabolism (PAM) and dominated endogenous metabolism. Ca. Competibacter (GAO) dominated the community following phosphorus-rich supernatant exclusion, with abundance increasing from 3.4% to 5.7%, accompanied by enhanced ED capacity (0.2 to 1.4 mg N/g VSS /h). The enriched subgroups (GB4, GB5) of Ca. Competibcater established a consistent nitrate cycle with anammox bacteria (AnAOB) through endogenous partial denitrification (EPD) at a ∆NO2--N/∆NH4+-N of 0.91±0.11, guaranteeing the maintenance of AnAOB abundance and performance. These results provide new insights into the flexibility of PNA for the energy-efficient treatment of low-strength ammonium wastewater.

Anammox upflow hybrid reactor: Nitrogen removal performance and potential for phosphorus recovery.

Echoing to the call of recovering high-value-added chemicals from wastewater and achieving carbon-neutral operation in wastewater treatment, an anammox upflow hybrid reactor was successfully applied for nitrogen removal, and the potential for phosphorus recovery was put forward. Moreover, the spatial pattern of removal capacities, and distribution of biomass and HAP precipitates were recognized and demonstrated as height-oriented. The intensity of HAP precipitates was highly consistent with the amount of anammox biomass and the relative abundance of the Candidatus Kuenenia, indicating that HAP formation was encouraged by the anammox reaction itself and heterogeneous nucleation induced by organic matters (proteins and polysaccharides). The fixed bed also played an important role in immobilizing the anammox biomass, secreted organic matrix, and HAP precipitates. This finding also provoked the thought that in the anammox process, HAP precipitation was more achievable, effective and practicable using the fixed-carrier system.

The phosphorus harvest from low-temperature mainstream wastewater through iron phosphate crystallization in a pilot-scale partial nitritation/anammox reactor.

The phosphorus harvest along nitrogen removal in the partial nitritation/anammox (PNA) reactor is promising for saving space and simplifying the management of mainstream wastewater treatment facilities. In this study, the phosphorus recovery from the low-temperature mainstream wastewater was explored through iron phosphate crystallization in a pilot-scale PNA reactor. With the COD-alleviated municipal wastewater as the influent, the ammonium concentration of about 50 mg/L and the phosphorus concentration ranged from 5.4 to 7.1 mg/L, under the temperature of 15 °C and the addition of external ferrous iron of 14 mg/L, the achieved nitrogen removal efficiency and the phosphorus removal efficiency were 37.6 % and 62.7 %, respectively. The good settleability of sludge indicated that the formed iron phosphate was well combined with the biomass. The quantitative analysis confirmed that the main iron phosphate in dry sludge was graftonite, and qualitative analysis confirmed that the equivalent of P2O5 content in the sludge was 5.8 %, which was suitable as fertilizer on agricultural land to realize the direct recycle of discharged phosphorus. In all, this study proposed a pioneering scheme to realize the nitrogen removal and phosphorus cycle in human society and given a meaningful reference for further research and application.

The responses of marine anammox bacteria-based microbiome to multi-antibiotic stress in mariculture wastewater treatment.

Saline mariculture wastewater containing multi-antibiotics poses a challenge to anaerobic ammonia oxidation (anammox) process. Herein, the halophilic marine anammox bacteria (MAB)-based microbiome was used for treating mariculture wastewater (35‰ salinity) under multi-antibiotics (enrofloxacin + oxytetracycline + sulfamethoxazole, EOS) stress. And the main focus of this study lies in the response of MAB-based microbiome against multi-antibiotics stress. It is found that MAB-based microbiome shows stable community structure and contributes high nitrogen removal efficiency (>90%) even under high stress of EOS (up to 4 mg·L-1). The relative abundance of main functional genus Candidatus Scalindua, responsible for anammox, had little change while controlling the influent EOS concentration within 4 mg·L-1, whereas, significantly decreased to 2.23% at EOS concentration of as high as 24 mg·L-1. As an alternative, antibiotic resistance bacteria (ARB) species Rheinheimera dominated the microbial community of MAB-based biological reactor under extremely high EOS stress (e.g. 24 mg·L-1 in influent). The response mechanism of MAB-based microbiome consists of extracellular and intracellular defenses with dependence of EOS concentration. For example, while EOS within 4 mg·L-1 in this study, most of the antibiotics were retained by extracellular polymeric substances (EPS) via adsorption; If increasing the EOS concentration to 8 and even 24 mg·L-1, part of antibiotics could intrude into the cells and cause the intracellular accumulation of antibiotic resistance genes (ARGs) (total abundance up to 2.44 × 10-1 copies/16S rRNA) for EOS response. These new understandings will facilitate the practical implementation of MAB-based bioprocess for saline nitrogen- and antibiotics-laden wastewater treatment.

Simultaneous nitrogen and phosphorus removal from municipal wastewater by Fe(III)/Fe(II) cycling mediated partial-denitrification/anammox.

The efficient removal of nitrogen and phosphorus remains challenging for traditional wastewater treatment. In this study, the feasibility for enhancing the partial-denitrification and anammox process by Fe (III) reduction coupled to anammox and nitrate-dependent Fe (II) oxidation was explored using municipal wastewater. The nitrogen removal efficiency increased from 75.5 % to 83.0 % by adding Fe (III). Batch tests showed that NH4+-N was first oxidized to N2 or NO2--N by Fe (III), then NO3--N was reduced to NO2--N and N2 by Fe (II), and finally, NO2--N was utilized by anammox. Furthermore, the performance of phosphorus removal improved by Fe addition and the removal efficiency increased to 78.7 %. High-throughput sequencing showed that the Fe-reducing bacteria Pseudomonas and Thiobacillus were successfully enriched. The abundance of anammox bacterial increased from 0.03 % to 0.22 % by multiple nitrite supply pathways. Fe addition presents a promising pathway for application in the anammox process.

Inorganic quantum dots - anammox consortia hybrid for stable nitrogen elimination under high-intensity solar-simulated irradiation.

External stimulus such as light irradiation is able to deteriorate intracellular redox homeostasis and induce photooxidative damage to non-photogenic bacteria. Exploiting effective strategies to help bacteria resisting infaust stress is meaningful for achieving a stable operation of biological treatment system. In this work, selenium-doped carbon quantum dots (Se-CQDs) were blended into anaerobic ammonia oxidation (anammox) bacteria and an inorganic nanoparticle-microbe hybrid was successfully fabricated to evaluate its nitrogen removal performance under solar-simulated irradiation. It was found that the specific anammox activity decreased by 29.7 ± 5.2% and reactive oxygen species (ROS) content increased by 134.8 ± 4.1% under 50,000 lux light. Sludge activity could be completely recovered under the optimum dosage of 0.42 mL·(g volatile suspended solid) -1 Se-CQDs. Hydroxyl radical (·OH) and superoxide anion radical (·O2-) were identified as the leading ROS inducing lipid peroxidation and antioxidase function detriment. Also, the structure of ladderane lipids located on anammoxosome was destroyed by ROS and functional genes abundances declined accordingly. Although cell surface coated Se-CQDs could absorb ultraviolet light and partially mitigated the photoinhibition, the direct scavenging of ROS by intracellular Se-CQDs primarily contributed to the cellular redox homeostasis, antioxidase activity recovery and sludge activity improvement. The findings of this work provide in-depth understanding the metabolic response mechanism of anammox consortia to light irradiation and might be valuable for a more stable and sustainable nitrogen removal technology, i.e., algal-bacterial symbiotic system, development.

Microbial driving mechanism for simultaneous removal of nitrogen and phosphorus in a pure anammox reactor under ferrous ion exposure.

The mechanisms of Fe2+ on nitrogen and phosphorus removal and functional bacterial competition in anammox systems was investigated. Under 0.12 mM Fe2+, the performance of nitrogen and phosphorus removal increased by 10.08 % and 151.91 %, respectively, compared with the control stage. Phosphorus removal was achieved through extracellular polymeric substance (EPS) induced biomineralization to form Fe-P minerals, and functional group COC in EPS played a critical role. T-EPSs was the major nucleation site due to it maintaining the supersaturated state (saturation index > 0) of Fe-P minerals for a long time. Population succession showed that Fe2+ weakened the competition between heterotrophic denitrifier (Denitrasoma) and anammox microbe (Candidatus Brocadia) for space and substrates, which was favorable for the enrichment of anammox biomass. Moreover, the variation in gene abundance (such as Hao, Cyt c, and Nir) indicated that Fe2+ improved electron behaviors (generation, transport, and consumption) during the nitrogen metabolism of anammox systems.

A novel process for anammox pretreatment of municipal wastewater: semi-partial nitrification, biological phosphorus removal and recovery.

Achieving simultaneous semi-partial nitrification and deep phosphorus removal is a preferred process technology for Anammox pretreatment. In this study, semi-partial nitrification combined with in-situ phosphorus recovery (PNPR) was used to treat municipal wastewater. The SRT conflict between the nitrification and phosphorus removal was resolved by in-situ phosphorus recovery every 20 cycles of Anaerobic/Oxid, and a supernatant with more than 10 times the influent phosphorus concentration was obtained, thus achieving bio-enhanced phosphorus removal and recovery with satisfactory semi-partial-nitrification effluent. Interestingly, the results showed that phosphorus removal and recovery process could improve the activity of AOB. The PNPR system's nitrite accumulation rate (NAR) and phosphorus removal rate (PRR) were more than 90% each, whereas the relative abundance of AOB and PAOs increased from 0.04% to 0.74% and from 0.25% to 0.70%, respectively (P < 0.01). Furthermore, on average, the NO2--Neff/NH4+-Neff value was 1.96, which laid the foundation for the subsequent anammox treatment.

Synergistic simultaneous endogenous partial denitrification/anammox (EPDA) and denitrifying dephosphatation for advanced nitrogen and phosphorus removal in a complete biofilm system.

To achieve simultaneous biological nitrogen and phosphorus removal from municipal wastewater, the endogenous partial denitrification/anammox (EPDA) was combined with denitrifying dephosphatation in a complete biofilm reactor. Advanced nitrogen and phosphorus removal were achieved with effluent total nitrogen (TN) and PO43--P concentrations of 7.77 ± 0.33 mg/L and 0.35 ± 0.10 mg/L, respectively. Anammox took a major role in the system, accounting for 76 ± 7% of nitrogen removal. 16S rRNA high-throughput sequencing results showed that the anammox bacteria co-existed with the denitrifying glycogen accumulating organisms (DGAOs) and the denitrifying phosphorus accumulating organisms (DPAOs). Anammox bacteria were mainly distributed in the inner layer, while DGAOs and DPAOs existed in the outer layer of EPDA biofilms. Furthermore, based on the EPDA biofilm system, a promising advanced nitrogen and phosphorus removal process was suggested to achieve lower requirements for energy and reagent consumption.