Anaerobic Ammonium Oxidation Process Research Articles

Novel insights into intrinsic mechanisms of magnetic field on long-term performance of anaerobic ammonium oxidation process

The performance of an anaerobic ammonium oxidation (anammox) reactor with the magnetic field of 40 mT was systematically investigated. The total nitrogen removal rate was enhanced by 16% compared with that of the control group. The enhancing mechanism was elucidated from the improved mass transfer efficiency, the complicated symbiotic interspecific relationship and the improved levels of functional genes. The magnetic field promoted formation of the loose anammox granular sludge and the homogeneous and well-connected porous structure to enhance the mass transfer. Consequently, Candidatus Brocadia predominated in the sludge with an increase in abundance of 13%. Network analysis showed that the positive interactions between Candidatus Brocadia and heterotrophic bacteria were strengthened, which established a more complicated stable microbial community. Moreover, the magnetic field increased the levels of hdh by 26% and hzs by 35% to promote the nitrogen metabolic process. These results provided novel insights into the magnetic field-enhanced anammox process.

Harnessing the Potential of Extracellular Polymeric Substances in Enhancing ANAMMOX Processes: Mechanisms, Strategies, and Perspectives

Anaerobic ammonium oxidation (ANAMMOX) has emerged as a promising sustainable nitrogen removal technology that offers significant advantages over conventional nitrification–denitrification processes, such as reduced energy consumption, a 60% reduction in oxygen demand, and a 90% reduction in sludge production. However, the practical application of ANAMMOX is hindered by several challenges, including the slow growth of ANAMMOX bacteria, long start-up periods, and high sensitivity to environmental disturbances. Recent studies have highlighted the crucial role of extracellular polymeric substances (EPSs) in the formation, activity, and stability of ANAMMOX biofilms and granules. An EPS is a complex mixture of high-molecular-weight polymers secreted by microorganisms, mainly composed of polysaccharides, proteins, nucleic acids, and lipids. The diverse physicochemical properties and functional groups of EPSs enable them to serve as a structural scaffold, protective barrier, sorption site, electron shuttle, and nutrient source for ANAMMOX bacteria. This review aims to provide an overview of the latest research progress on harnessing the potential of EPSs to enhance the ANAMMOX process. The characteristics, compositions, and extraction methods of ANAMMOX-derived EPSs are summarized. The mechanisms of how EPSs facilitate the enrichment, immobilization, aggregation, and adaptation of ANAMMOX bacteria are elucidated. The strategies and effects of EPS supplementation on improving the performance and robustness of ANAMMOX reactors under various stresses are critically reviewed. The challenges and future perspectives of the EPS-mediated optimization of the ANAMMOX process are also discussed. This review sheds new light on exploiting EPSs as a renewable bioresource to develop more efficient and stable ANAMMOX applications for sustainable wastewater treatment.

Regulation of nitrogen removal pathways in sulfuric autotrophic denitrification combined with ANAMMOX

The process of sulfuric autotrophic denitrification combined with anaerobic ammonia oxidation (ANAMMOX) has attracted attention because of its efficient N removal capacity, low sludge yield and low operating cost. However, the NO2−-N generated by the sulfuric autotrophic partial denitrification (SAPDN) easily continues conversion to N2 in the coupled process. Thus, ANAMMOX cannot occur in the absence of NO2−-N. In this study, the regulation of N removal pathways was studied in the combined system. The N removal performance was evaluated for treating the industrial wastewater in NO3−-N and NH4+-N with the sulfuric autotrophic denitrification combined with the ANAMMOX process. The effects of influent nitrogen load, concentration ratio of NO3−-N/NH4+-N as well as S/N mass ratio on the transformation of nitrogen removal pathway were studied. SAPDN combined with ANAMMOX became the primary nitrogen removal pathway due to regulating the high nitrogen load of 0.83–0.96 kg/(m3·d), average ratio of NO3−-N/NH4+-N ranging from 1.38 to 1.70 in the influent and a low S/N ratio of 1.5. Moreover, the microbial analysis results indicated that Thiobacillus and Candidatus Brocadia became the main functional microbes when the primary nitrogen removal pathway was SAPDN combined with ANAMMOX.

Carbon emission calculation of three biological nitrogen removal processes: A case of rare earth element tailings wastewater in Southern Jiangxi, China

Global warming has been caused by greenhouse gases (GHGs) released by human activity. Traditional sewage treatment uses much energy to achieve discharge limits in the water treatment industry, producing many GHG emissions. In this study, the emission factor approach and mass balance method are used to calculate the carbon emission equivalent of nitrification and denitrification, anaerobic ammonia oxidation, and microalgae assimilation using a rare earth tailings (REEs) wastewater example. The results show that, among the three denitrification processes, the traditional nitrification and denitrification method has the greatest carbon emission equivalent (16.23 kgCO2/kgN). Direct causes include N2O emissions, which account for 48 % of the total carbon emission equivalent. The anaerobic ammonia oxidation process produces the carbon dioxide equivalent of 0.02 kgCO2/kgN. Carbon emission comparable to microalgal assimilation is −3.23 kgCO2/kgN. The anaerobic ammonia oxidation process can thus greatly reduce carbon emissions when compared to the conventional nitrification and denitrification processes, whilst microalgae assimilation can result in negative carbon emissions.

Discrepant effects of undissolved and dissolved fractions of biochar on the nitrogen removal performance of anaerobic ammonium oxidation process

Biochar has attracted increasingly attention in the biological wastewater treatment owing to its unique environmental behavior. However, the differential effects of undissolved and dissolved fractions of biochar (hereafter named as UDB and DB, respectively) on the anaerobic ammonium oxidation (anammox) process remain unclear. The study evaluated the nitrogen removal performance of anammox system in response to UDB and DB prepared at different pyrolysis temperatures (300 °C, 550 °C and 800 °C). The DB800-amended group displayed superior nitrogen removal performance with the total nitrogen removal efficiency (TNRE) of 92.8 ± 1.4 %. Compared with CK, TNRE was changed by +5.0 % (UDB300), +0.4 % (UDB550) and −22.3 % (UDB800) for UDB-amended groups, and by −16.0 % (DB300), −9.4 % (DB550), and +9.2 % (DB800) for DB-amended groups, respectively. The differences were closely related to the physicochemical properties of UDB and DB. Particularly, dissolved organic matter released by DB were tracked at molecular level, which revealed that CHO occupied the highest proportion of 70.3 % and lignin-like and carboxylic rich alicyclic molecules were dominant in DB800. This study would facilitate the understanding of the differences between the nitrogen removal performance of UDB and DB-amended anammox, thus providing deeper insights for the comprehensive assessment of the impacts of biochar on the anammox process.

Research progress of anaerobic ammonium oxidation (Anammox) process based on integrated fixed-film activated sludge (IFAS).

The integrated fixed-film activated sludge (IFAS) process is considered one of the cutting-edge solutions to the traditional wastewater treatment challenges, allowing suspended sludge and attached biofilm to grow in the same system. In addition, the coupling of IFAS with anaerobic ammonium oxidation (Anammox) can further improve the efficiency of biological denitrification. This paper summarises the research progress of IFAS coupled with the anammox process, including partial nitrification anammox, simultaneous partial nitrification anammox and denitrification, and partial denitrification anammox technologies, and describes the factors that limit the development of related processes. The effects of dissolved oxygen, influent carbon source, sludge retention time, temperature, microbial community, and nitrite-oxidising bacteria inhibition methods on the anammox of IFAS are presented. At the same time, this paper gives an outlook on future research focus and engineering practice direction of the process.

Deciphering behaviors of 6:2 chlorinated polyfluorinated ether sulfonate (alternative-PFOS) on anammox processes: Nitrogen removal efficiency and microbial adaptability

This study investigates the behaviors and effects of F-53B, an alternative to perfluorooctane sulfonate on anaerobic ammonium oxidation (anammox) processes. Results showed that the nitrogen removal efficiency (NRE) reached 83.8 % at a F-53B concentration of 0.5 mg·L−1, while NRE decreased to 66.9 % with 5 mg·L−1 of F-53B. The defluorination rates of 17.8 % (0.5 mg·L−1) and 9.3 % (5 mg·L−1) were observed, respectively, suggesting the occurrence of F-53B degradation. The relative abundance of Ca. Kuenenia decreased from 26.1 % to 16.2 % with the F-53B concentration increasing from 0.5 mg·L−1 to 5 mg·L−1. Meanwhile, Denitratisoma was selectively enriched with a relative abundance of 40.7 % at an F-53B concentration of 0.5 mg·L−1. Ca. Kuenenia could reduce reactive oxygen species induced by F-53B to maintain the balance of oxidative stress. This study gains insight into the behaviors and metabolic mechanisms of F-53B in anammox consortia, suggesting the feasibility of anammox processes for industrial wastewater.

Insight into microbial adaptability in continuous flow anaerobic ammonium oxidation process for low-strength sewage treatment

Organic matter concentration is a critical factor influencing the adaptability of anaerobic ammonium oxidation (anammox) bacteria to low-strength sewage treatment. To address this challenge and achieve stable anammox activity, a micro-aeration partial nitrification-anammox process was developed for continuous-flow municipal sewage treatment. Under limited ammonium conditions, the effective utilization of organics in denitrification promoted the stable accumulation of nitrite and enhanced anammox activity. This, in turn, led to enhanced nitrogen removal efficiency, reaching approximately 87.7%. During the start-up phase, the protein content of extracellular polymeric substances (EPS) increased. This enhanced EPS intensified the inhibitory effect of denitrifying bacteria (DNB) on nitrite-oxidizing bacteria through competition for nitrite, thereby facilitating the proliferation of anammox bacteria (AnAOB). Additionally, several types of DNB capable of utilizing slowly biodegradable organics contributed to the adaptability of AnAOB. These findings provide valuable insights for ensuring efficient anammox performance and robust nitrogen removal in the treatment of low-strength sewage.

Analyzing the nitrogen removal performance and cold adaptation mechanism of immobilized cold-acclimation ANAMMOX granules at low temperatures.

To improve the treatment performance of anaerobic ammonium oxidation (ANAMMOX) processes at low temperatures, the immobilized cold-acclimation ANAMMOX granules (R3) were prepared and their low-temperature nitrogen removal ability as well as the cold adaptation mechanism were analyzed. The results indicated that the total inorganic nitrogen (TIN) removal efficiency of R3 was significantly higher than that of R2 (cold-acclimation granules without immobilization) and R1 (common granules), especially at 11 ± 2 and 7±2°C (68% and 54%). These were attributed to the remarkable biomass retention capacity of R3, high up to 4.3-4.9mg/gVSS even at 5-18°C. Besides, higher protein (PN) content of tightly bound extracellular polymeric substances (TB-EPS) also facilitated microbial aggregation in R3. Meanwhile, R3 granules retained higher ANAMMOX activity and heme c content at 5-25°C. The original dominant ANAMMOX genus (Candidatus Kuenenia) in R3 kept higher abundance (49%-57%) at 23 ±2 and 16 ±2°C, whereas Candidatus Brocadia became the dominant ANAMMOX genus (25%-32%) in R3 at 11 ±2 and 7±2°C. Notably, different ANAMMOX genera in R3 may adapt to cold environment by regulating the expression of cold-stress proteins (CspA, CspB, PpiD, and UspA). PRACTITIONER POINTS: Immobilized cold-acclimation ANAMMOX granules showed higher nitrogen removal efficiency at 23°C → 5°C. Immobilization method effectively retained biomass (Candidatus Kuenenia and Candidatus Brocadia). Immobilization facilitated TB-EPS release and biological aggregation in cold-acclimation granules. Expression of cold-stress proteins in immobilized cold-acclimation granules was more active.

Insights into nitrogen removal from leather wastewater by anaerobic ammonium oxidation process: Performance and microbial structure

This study aimed to assess the suitability of anaerobic ammonium oxidation (anammox) process for leather wastewater treatment using two up-flow anaerobic sludge blanket bioreactors by pumping synthetic and real wastewater (named SR and RR), respectively. Both reactors were operated steadily with an average nitrogen removal efficiency of 94–96 % at chemical oxygen demand (COD) level of 100–200 mg·L−1, which were rapidly deteriorated as the COD level increased to 300 mg·L−1 even if at the same COD to total nitrogen (TN) (COD/TN) ratio. The reactor performances could restore after pumping with synthetic wastewater, and then continued to effectively treat leather wastewater. Furthermore, the relative abundance of Candidatus Kuenenia in reactor RR was increased by 34.8 % compared with the reactor SR on day 120. The results revealed that the anammox process can be used for the leather wastewater treatment, thus broadening the engineering application of anammox-based process in industrial wastewater.

Diversity of Anammox Bacteria from Landfill Treatment Plant Sludge in Tropical Area

The anaerobic ammonium oxidation (anammox) process is known as the warm process. Tropical areas have an advantage due to their consistent temperature throughout the year. This study analyzed the diversity of anammox bacteria in the tropical area using leachate sludge from a landfill as an inoculum in a filter bioreactor (FtBR) and observed nitrogen removal performance. Ammonium and nitrite concentrations of 70, 150, and 200 mg-N/L were delivered into the reactor continuously with hydraulic retention time (HRT) of 24 h and 12 h and run for 131 days at ambient tropical temperature (25–28 °C). High performance achieved with nitrogen removal rate (NRR), nitrogen removal efficiency (NRE), and ammonium conversion efficiency (ACE) were 0.866 kg-N/m3.d, 99.19%, and 98.90%, respectively. The cultivated leachate sludge could perform an anammox process with four anammox species, Candidatus Brocadia fulgida, Candidatus Brocadia sapporoensis, Candidatus Brocadia sp uncultured, Candidatus Jettenia sp with abundance 6.52%, 13.82%, 0.77%, and 0.69%, respectively. These findings contribute to the advancement of biotechnology in wastewater treatment, particularly in tropical countries, and highlight the potential for highly cost-effective technology.

Impacts of crude glycerol on anaerobic ammonium oxidation (Anammox) process in wastewater treatment

This work investigated the impact of a waste-derived carbon source, crude glycerol (CG), on Anammox. Batch bioassays were conducted to identify inhibitory component(s) in CG, and the relationship between Anammox activity and the concentration of CG, pure glycerol, and methanol were assessed. The results showed that the half-maximal inhibitory concentration of CG and methanol are 434.5 ± 51.8 and 143.0 ± 19.6 mg chemical oxygen demand (COD) L-1, respectively, while pure glycerol at 0–2283 mg COD L-1 had no significant adverse effect on Anammox. The results suggested methanol is the major inhibitor in CG via a non-competitive inhibition mechanism. COD/total inorganic nitrogen ratio of > 1.3 was observed to cause a significant Anammox inhibition (>20 %), especially at low substrate level. These results are valuable for evaluating the feasibility of using CG for nitrogen removal in water resource recovery facilities, promoting sustainable development.

Efficient nitrite accumulation in partial sulfide autotrophic denitrification (PSAD) system: insights of S/N ratio, pH and temperature

ABSTRACT To provide the necessary nitrite for the Anaerobic Ammonium Oxidation (ANAMMOX) process, the effect of nitrite accumulation in the partial sulfide autotrophic denitrification (PSAD) process was investigated using an SBR reactor. The results revealed that the effectiveness of nitrate removal was unsatisfactory when the S/N ratio (mol/mol) fell below 0.6. The optimal conditions for nitrate removal and nitrite accumulation were achieved within the S/N ratio range of 0.7-0.8, resulting in an average Nitrate Removal Efficiency (NRE) of 95.84%±4.89% and a Nitrite Accumulation Rate (NAR) of 75.31%±6.61%, respectively. It was observed that the nitrate reduction rate was three times faster than that of nitrite reduction during a typical cycle test. Furthermore, batch tests were conducted to assess the influence of pH and temperature conditions. In the pH tests, it became evident that the PSAD process performed more effectively in alkaline environment. The highest levels of nitrate removal and nitrite accumulation were achieved at an initial pH of 8.5, resulting in a NRE of 98.30%±1.93% and a NAR of 85.83%±0.47%, respectively. In the temperature tests, the most favourable outcomes for nitrate removal and nitrite accumulation were observed at 22±1 ℃, with a NRE of 100.00% and a NAR of 81.03%±1.64%, respectively. Moreover, a comparative analysis of 16S rRNA sequencing results between the raw sludge and the sulfide-enriched culture sludge sample showed that Proteobacteria (49.51%) remained the dominant phylum, with Thiobacillus (24.72%), Prosthecobacter (2.55%), Brevundimonas (2.31%) and Ignavibacterium (2.04%) emerging as the dominant genera, assuming the good nitrogen performance of the system.

Development of partial denitrification process in upflow-anaerobic sludge blanket and effect of electric field on partial denitrification performance

Partial denitrification (PD) is an alternative to providing NO2– for the anaerobic ammonium oxidation (anammox) process. In this study, three upflow anaerobic sludge blankets (UASB) were used to investigate the effect of an external electric field on PD performance. The results indicated that the maximum nitrite transformation ratio (NTR) reached 76.3 %, with an average NTR of 54.1 %, in the presence of external electric field, whereas the average NTR of the control was only 49.8 %. The fitted maximum specific nitrate reduction rates of PD1, PD2, and PD3 were 83.7, 90.5, and 92.3 mg N g−1VSS h−1, respectively, according to the Haldane model analysis. Microbial community analysis demonstrated that the abundance of Thauera, Comamonas, and Accumulibacter increased with electric assistance. In summary, UASB reactor with electrodes set in the upper region was most feasible for the stable PD process, providing an alternative for developing a coupled PD-anammox process.

Rapid start-up strategy and microbial population evolution of anaerobic ammonia oxidation biofilm process for low-strength wastewater treatment

The implementation of the anaerobic ammonium oxidation (anammox) process in treating low-strength wastewater is limited by the difficulty in enriching anammox bacteria (AnAOB). Here, the first enrichment of AnAOB at a high nitrogen (N) loading rate (NLR) as a strategy was proposed to achieve the rapid start-up of the anammox biofilm process treating low-strength wastewater. The long-term stability of the anammox biofilm process after start-up operating at a low NLR of 0.2–0.4 kg N/(m3⋅d) was evaluated. Results showed that the N removal efficiency was up to 75 % under a low NLR of 0.2 kg N/(m3⋅d) condition. Low-strength organic matter promoted the metabolic coupling between partial denitrifying bacteria (PDB) and AnAOB. The genus Candidatus Brocadia as AnAOB (18 %-27 %) can coexist with Limnobacter (PDB, 9 %-12 %) for efficient N removal. This study offers a rapid start-up strategy of anammox biofilm process in treating low-strength wastewater.

Waste iron scraps promote anammox bacteria to resist inorganic carbon limitation

The anaerobic ammonium oxidation (anammox) process is adversely affected by the limitation of inorganic carbon (IC). In this research, a new technique was introduced to assist anammox biomass in counteracting the adverse effects of IC limitation by incorporating waste iron scraps (WIS), a cheap and easily accessible byproduct of lathe cutting. Results demonstrated that reducing the influent IC/TN ratio from 0.08–0.09 to 0.04 resulted in a 20 % decrease in the nitrogen removal rate (NRR) for the control reactor, with an average specific anammox activity (SAA) of 0.65 g N/g VSS/day. Nevertheless, the performance of the WIS-assisted anammox reactor remained robust despite the reduction in IC supply. In fact, the NRR and SAA of the WIS-assisted reactor exhibited substantial improvements, reaching approximately 1.86 kg/(m3·day) and 0.98 g N/g VSS/day, respectively. These values surpassed those achieved by the control reactor by approximately 39 % and 51 %, respectively. The microbial analysis confirmed that the WIS addition significantly stimulated the proliferation of anammox bacteria (dominated by Candidatus Kuenenia) under IC limitation. The anammox gene abundances in the WIS-assisted anammox reactor were 3–4 times higher than those in the control reactor. Functional genes prediction based on the KEGG database revealed that the addition of WIS significantly enhanced the relative abundances of genes associated with nitrogen metabolism, IC fixation, and central carbon metabolism. Together, the results suggested that WIS promoted carbon dioxide fixation of anammox species to resist IC limitation. This study provided a promising approach for effectively treating high ammonium-strength wastewater using anammox under IC limitation.

Wastewater Treatment System Optimization for Sustainable Operation of the SHARON–Anammox Process under Varying Carbon/Nitrogen Loadings

Partial nitritation (PN) coupled with the anaerobic ammonium oxidation (Anammox) process has improved ammonium removal in wastewater treatment plants (WWTPs). The operation conditions of this process, i.e., the dissolved oxygen (DO) and the influent ammonium and nitrite concentrations, drive the process to an equilibrium to suppress nitrite-oxidizing bacteria and achieve a proper nitrite over ammonium (NO2/NH4) ratio. This study aimed to implement a set of control strategies in a WWTP model BSM2-SHAMX, combining PN in a single reactor system for high-activity ammonia removal over nitrite (SHARON) to an Anammox reactor, using proportional–integrative–derivative (PID) control and model predictive control (MPC) in a cascade. For correct coupling, the PN should maintain an output NO2/NH4 ratio between 1 and 1.3, suitable for the Anammox process. In the cascade controller feedback loop, the primary control loop controls the NO2/NH4 ratio through the DO concentration from the secondary control loop, which guarantees better effluent nitrogen removal. The performance of the plant was assessed by evaluating the control strategies with different influent carbon/nitrogen (C/N) loadings. The study results showed that the MPC controllers provided better results, with an improvement of 36% in the operational cost compared to the base case with a cost around 26,000 EUR/d, and better nitrogen removal surpassing 90% removal, 10% more than the base case.

Quantitative effect of adding percentages of anammox granules on the start-up process and microbial community analysis

The anaerobic ammonium oxidation (anammox) process is challenging due to its long start-up duration and high demand for mature anammox seed sludge. However, adding a small amount of anammox sludge to the inoculum can be a reasonable solution. This study investigated the effect of adding percentage of anammox granules (0, 1, 2, 4, and 8%) in the seed sludge on the anammox start-up process. The anammox process was achieved in all five reactors after 55, 6, 5, 3 and 0 days. Increasing the adding percentage effectively shortened the duration of lag phase and cell lysis, but had little effect on the final nitrogen removal performance, except for 4% adding percentage. Families of Brocadiaceae, Burkholderiaceae, Ignavibacteriaceae, SJA-28, and Rhodocyclaceae were dominant, with a core microbiota of eight operational taxonomic unites (OTUs), and Candidatus Brocadia fulgida became the dominant anammox species. Seven synergistic members with anammox bacteria were identified by correlation network analysis. Major potential functional groups involved in C and N cycle were also observed by FAPROTAX. Together with the qPCR and sequencing results, it was suggested that more than 2% of adding percentages would result in a short lag phase, rapid growth rate in elevation stage, high final performances, and anammox bacteria abundance comparable to that in the anammox seed sludge. This crucial finding indicated the feasibility of economical and rapid start-up of the anammox process with a minimum amount of anammox seed sludge.

Effect of Fe2+ on ANAMMOX Granular Sludge Cultured in a Biased Acidic Influent and Dynamic Environment

A continuous stirred tank reactor (CSTR) was utilized in this study to enrich and cultivate anaerobic ammonia oxidation process (ANAMMOX) granular sludge by gradually decreasing its pH, and to investigate the effects of different concentrations of ferrous ions (Fe2+) on the activity of ANAMMOX granular sludge cultivated under biased acidic conditions. The final nitrogen removal of ANAMMOX deteriorated at pH 6.30–6.50 after 220 days of continuous operation, but the nitrogen removal of ANAMMOX was favorable at pH 6.50–7.00. This indicates that a slightly acidic environment (pH = 6.50–7.00) promotes the activity of ANAMMOX, but the pH should not be too low (pH = 6.30–6.50). In the reactor, Candidatus Kuenenia was consistently the dominant ANAMMOX genus and its abundance declined from 11.70% on day 1 to 10.44% on day 220. As Fe2+ concentrations were increased (10, 20, 30 mg/L) in ANAMMOX granular sludge cultured in an acidic environment, the nitrogen removal effects gradually increased. In addition, with the increase in Fe2+ concentrations, the total nitrogen removal load (NRL) in the reactor was increased from 1.16 kg/(m3/d) to 1.42 kg/(m3/d). Increases in Fe2+ concentration did not result in inhibition of ANAMMOX, which may be attributed to the morphology of sludge and the shape of the reactor. As a result of the present study, new insights were gained into the physiological characteristics of ANAMMOX in an acidic environment over the long term, and how Fe2+ affects its ability to remove nitrogen from the environment.

Use of soluble iron salt as an electron acceptor to provide Fe(III) for Feammox process: Ammonium removal performance and mechanism

Fe(III) reduction coupled with anaerobic ammonium oxidation (Feammox) is a newly discovered biological ammonium removal technology. Compared to anaerobic ammonium oxidation (anammox) process, Feammox does not require NO2−-N as a reaction substrate and is more tolerant to heavy metals. In this study, soluble iron salts FeCl3 was added to anaerobic sequencing batch reactor (ASBR) to start Feammox process. The effects of Fe/N, pH and temperature on the Feammox performance were investigated. Optimal working conditions were determined by combining the variation law of nitrogen and iron elements. The results showed that the best NH4+-N removal efficiency when Fe/N, pH and temperature were 0.15, 6.5 and 30 °C, respectively. The modified Boltzmann model fitted better in Feammox kinetic analysis (R2 = 0.9789), which could clearly explain that the degradation of substrate and the generation of product. Microbial analysis showed that in addition to the typical iron-reducing bacteria, the dominant genus Candidatus_Brocadia of anammox was also enriched in the system with relative abundance of 3.16 %, implying that both Feammox and anammox contributed to the transformation of NH4+-N. Overall, optimizing the operating conditions of Feammox is important significance to maintain the stability of NH4+-N removal performance and provide theoretical basis for constructing a novel wastewater treatment process.