Novel MBR_based main stream biological nutrient removal process: high performance and microbial community

For municipal wastewater treatment, main stream biological nutrient removal (BNR) process is becoming more and more important. This lab-scale study, novel MBR_based BNR processes (named A₂N-MBR and A₂NO-MBR) were built. Comparison of the COD removal, results obtained demonstrated that COD removal efficiencies were almost the same in three processes, with effluent concentration all bellowed 30 mg L^?1. However, the two-sludge systems (A₂N-MBR and A₂NO-MBR) had an obvious advantage over the A²/O for denitrification and phosphorus removal, with the average TP removal rates of 91.20, 98.05% and TN removal rates of 73.00, 79.49%, respectively, higher than that of 86.45 and 61.60% in A²/O process. Illumina Miseq sequencing revealed that Candidatus_Accumulibacter, which is capable of using nitrate as an electron acceptor for phosphorus and nitrogen removal simultaneously, was the dominant phylum in both A₂N-MBR and A₂NO-MBR process, accounting for 28.74 and 23.98%, respectively. Distinguishingly, major organism groups related to nitrogen and phosphorus removal in A²/O system were Anaerolineaceae_uncultured, Saprospiraceae_uncultured and Thauera, with proportions of 11.31, 8.56 and 5.00%, respectively. Hence, the diversity of dominant PAOs group was likely responsible for the difference in nitrogen and phosphorus removal in the three processes.     

Simultaneous nitrogen and carbon removal in a packed A/O reactor: effect of C/N ratio on microbial community structure

In this research, a novel packed anoxic/oxic moving bed biofilm reactor (MBBR) was established to achieve high-organic matter removal rates, despite the carbon/nitrogen (C/N) ratio of 2.7–5.1 in the influent. Simultaneous nitrification–denitrification (SND) was investigated under a long sludge retention time of 104 days. The system exhibited excellent performance in pollutant removal, with chemical oxygen demand and total nitrogen (TN) enhanced to 93.6–97.4% and 34.4–60%, respectively. Under low C/N conditions, the nitrogen removal process of A/O MBBR system was mainly achieved by anaerobic denitrification. The increase of C/N ratio enhanced SND rate of the aerobic section, where dissolved oxygen was maintained at the range of 4–6 mg/L, and resulted in higher TN removal efficiency. The microbial composition and structures were analyzed utilizing the MiSeq Illumina sequencing technique. High-throughput pyrosequencing results indicated that the dominant microorganisms were Proteobacteria and Bacteroidetes at the phylum level, which contributes to the removal of organics matters. In the aerobic section, abundances of Nitrospirae (1.12–29.33%), Burkholderiales (2.15–21.38%), and Sphingobacteriales (2.92–11.67%) rose with increasing C/N ratio in the influent, this proved that SND did occur in the aerobic zone. As the C/N ratio of influent increased, the SND phenomenon in the aerobic zone of the system is the main mechanism for greatly improving the removal rate of TN in the aerobic section. The C/N ratio in the aerobic zone is not required to be high to exhibit good TN removal performance. When C/NH₄⁺ and C/TN in the aerobic zone were higher than 2.29 and 1.77, respectively, TN removal efficiency was higher than 60%, which means that carbon sources added to the reactor could be saved. This study would be vital for a better understanding of microbial structures within a packed A/O MBBR and the development of cost-efficient strategies for the treatment of low C/N wastewater.

     

Impact of COD/N ratio on nitrous oxide emission from microcosm wetlands and their performance in removing nitrogen from wastewater

Constructed wetlands (CWs) are considered to be important sources of nitrous oxide (N₂O). In order to investigate the effect of influent COD/N ratio on N₂O emission and control excess emission from nitrogen removal, free water surface microcosm wetlands were used and fed with different influent. In addition, the transformation of nitrogen was examined for better understanding of the mechanism of N₂O production under different operating COD/N ratios. It was found that N₂O emission and the performance of microcosm wetlands were significantly affected by COD/N ratio of wastewater influent. Strong relationships exist between N₂O production rate and nitrite (r = 0.421, p < 0.01). During denitrification process, DO concentration crucially influences N₂O production rate. An optimal influent COD/N ratio was obtained by adjusting external carbon sources for most effective N₂O emission control and best performance of the CWs in nitrogen removal from wastewater. It is concluded that under the operating condition of COD/N ratio = 5, total N₂O emission is minimum and the microcosm wetland is most effective in wastewater nitrogen removal.     

Comparison of the Bacterial Communities in Anaerobic, Anoxic, and Oxic Chambers of a Pilot A2O Process Using Pyrosequencing Analysis

A₂O process is a sequential wastewater treatment process that uses anaerobic, anoxic, and oxic chambers for nitrogen and phosphorus removal. In this study, the bacterial communities among these chambers were compared, and the diversity of the bacteria involved in nitrogen and phosphorus removal was surveyed. A pilot-scale A₂O process (50 m³ day^?1) was operated for more than 6 months, and bacterial 16S rRNA gene diversity was analyzed using pyrosequencing. A total of 7,447 bacterial sequence reads were obtained from anaerobic (1,546), anoxic (2,158), and oxic (3,743) chambers. Even though there were differences in the atmospheric condition and functionality, no prominent differences could be found in the bacterial community of the three chambers of the pilot A₂O process. All sequence reads, which were taxonomically analyzed using the Eztaxon-e database, were assigned into 638 approved or tentative genera. Among them, about 72.2 % of the taxa were contained in the phyla Proteobacteria and Bacteroidetes. Phosphate-accumulating bacteria, Candidatus Accumulibacter phosphatis, and two other Accumulibacter were found to constitute 3.1 % of the identified genera. Ammonia-oxidizing bacteria, Nitrosomonas oligotropha, and four other phylotypes in the same family, Nitrosomonadaceae, constituted 0.2 and 0.9 %, respectively. Nitrite-oxidizing bacteria, Nitrospira defluvii, and other three phylotypes in the same family, Nitrospiraceae, constituted 2.5 and 0.1 %, respectively. In addition, Dokdonella and a phylotype of the phylum Chloroflexi, function in nitrogen and/or phosphate removal of which have not been reported in the A₂O process, constituted the first and third composition among genera at 4.3 and 3.8 %, respectively.     

Nonlinear nitrous oxide (N2O) response to nitrogen fertilizer in on‐farm corn crops of the US Midwest

Row‐crop agriculture is a major source of nitrous oxide (N₂O) globally, and results from recent field experiments suggest that significant decreases in N₂O emissions may be possible by decreasing nitrogen (N) fertilizer inputs without affecting economic return from grain yield. We tested this hypothesis on five commercially farmed fields in Michigan, USA planted with corn in 2007 and 2008. Six rates of N fertilizer (0–225 kg N ha^?1) were broadcast and incorporated before planting, as per local practice. Across all sites and years, increases in N₂O flux were best described by a nonlinear, exponentially increasing response to increasing N rate. N₂O emission factors per unit of N applied ranged from 0.6% to 1.5% and increased with increasing N application across all sites and years, especially at N rates above those required for maximum crop yield. At the two N fertilizer rates above those recommended for maximum economic return (135 kg N ha^?1), average N₂O fluxes were 43% (18 g N₂O–N ha^?1 day^?1) and 115% (26 g N₂O–N ha^?1 day^?1) higher than were fluxes at the recommended rate, respectively. The maximum return to nitrogen rate of 154 kg N ha^?1 yielded an average 8.3 Mg grain ha^?1. Our study shows the potential to lower agricultural N₂O fluxes within a range of N fertilization that does not affect economic return from grain yield.     

Simultaneous nitrification and denitrification with different mixed nitrogen loads by a hypothermia aerobic bacterium

Microorganism with simultaneous nitrification and denitrification ability plays a significant role in nitrogen removal process, especially in the eutrophic waters with excessive nitrogen loads. The nitrogen removal capacity of microorganism may suffer from low temperature or nitrite nitrogen source. In this study, a hypothermia aerobic nitrite-denitrifying bacterium, Pseudomonas tolaasii strain Y-11, was selected to determine the simultaneous nitrification and denitrification ability with mixed nitrogen source at 15 °C. The sole nitrogen removal efficiencies of strain Y-11 in simulated wastewater were obtained. After 24 h of incubation at 15 °C, the ammonium nitrogen fell below the detection limit from an initial value of 10.99 mg/L. Approximately 88.0 ± 0.33% of nitrate nitrogen was removed with the initial concentration of 11.78 mg/L and the nitrite nitrogen was not detected with the initial concentration of 10.75 mg/L after 48 h of incubation at 15 °C. Additionally, the simultaneous nitrification and denitrification nitrogen removal ability of P. tolaasii strain Y-11 was evaluated using low concentration of mixed NH₄⁺-N and NO₃^?–N/NO₂^?–N (about 5 mg/L-N each) and high concentration of mixed NH₄⁺–N and NO₃^?–N/NO₂^?–N (about 100 mg/L-N each). There was no nitrite nitrogen accumulation at the time of evaluation. The results demonstrated that P. tolaasii strain Y-11 had higher simultaneous nitrification and denitrification capacity with low concentration of mixed inorganic nitrogen sources and may be applied in low temperature wastewater treatment.     

黄河上游灌区稻田N2O排放特征

黄河上游灌区稻田高产区过量施肥现象十分突出,氮肥过量施用引起土壤氮素盈余,导致N₂O排放量增大,由此引起的温室效应引起广泛关注。采用静态箱-气相色谱法研究黄河上游灌区稻田不同施肥处理下N₂O排放特征。试验设置5个施肥处理,包括常规氮肥300 kg/hm²下单施尿素和有机肥配施2个处理,分别用N300和N300-OM代表;优化氮肥240 kg/hm²下单施尿素和有机肥配施2个处理,分别用N240和N240-OM代表;对照不施氮肥用N0代表。试验结果得出,灌区水稻生长季稻田土壤N₂O排放主要集中在水稻分蘖前及水稻生长的中后期,稻田氮肥施用、灌水及土壤温度的变化对N₂O排放通量影响较大,不同处理水稻各生育阶段N₂O累积排放量与稻田土壤耕层NO^-₃-N含量动态变化显著相关。稻田N₂O排放不是黄河上游灌区稻田氮素损失的主要途径,但灌区稻田N₂O排放的增温潜势较大;稻田氮肥过量施用会显著增加N₂O排放量,在相同氮素水平下,有机肥配施会显著增加稻田土壤N₂O的排放量(P＜0.01)。优化施氮能有效减少灌区稻田水稻生长季N₂O排放量。稻田不同处理的水稻整个生长季土壤N₂O排放总量为2.69-3.87 kg/hm²,肥料氮通过N₂O排放损失的百分率仅为0.43%-0.64%。在灌区习惯灌水和高氮肥300 kg/hm²时,N300-OM处理的稻田N₂O排放量达3.87 kg/hm²,在100 a时间尺度上的全球增温潜势(GWPs)为20.76×10⁷ kg CO₂/hm²;优化施氮240 kg/hm²水平下,N240和N240-OM处理的N₂O累计排放量较N300-OM处理,分别降低了1.18 kg/hm²和0.57 kg/hm²,在100 a尺度上每年由稻田N₂O排放引起的GWPs分别降低了6.33×10⁷ kg CO₂/hm²和3.06×10⁷ kg CO₂/hm²。     

Evaluation of pilot-scale modified A2O processes for the removal of nitrogen compounds from sewage

In this study, pilot-scale experiments for the removal of nitrogen from sewage obtained from a county Y sewer system were performed using modified A₂O processes. Using this approach, the total amount of nitrogen discharged during denitrification of the influent was average 38.6 mg/L and a level of average 10.8 mg/L was maintained throughout the denitrification process, which resulted in an average removal efficiency that was greater than 72%. The nitrogen components in the effluent water consisted of 22% ammonia nitrogen, 6% nitrite nitrogen and 72% nitrate nitrogen, reaching a nitrification efficiency of 94%. In conclusion, since these advanced treatment methods, which involve modified A₂O processes, were successfully employed to remove nitrogen from sewage discharge, they hold promise for wide spread use by treatment plants.     

One-stage partial nitritation/anammox at 15 °C on pretreated sewage: feasibility demonstration at lab-scale

Energy-positive sewage treatment can be achieved by implementation of oxygen-limited autotrophic nitrification/denitrification (OLAND) in the main water line, as the latter does not require organic carbon and therefore allows maximum energy recovery through anaerobic digestion of organics. To test the feasibility of mainstream OLAND, the effect of a gradual temperature decrease from 29 to 15 °C and a chemical oxygen demand (COD)/N increase from 0 to 2 was tested in an OLAND rotating biological contactor operating at 55–60 mg NH₄ ⁺–N?L^?1 and a hydraulic retention time of 1 h. Moreover, the effect of the operational conditions and feeding strategies on the reactor cycle balances, including NO and N₂O emissions were studied in detail. This study showed for the first time that total nitrogen removal rates of 0.5 g N?L^?1?day^?1 can be maintained when decreasing the temperature from 29 to 15 °C and when low nitrogen concentration and moderate COD levels are treated. Nitrite accumulation together with elevated NO and N₂O emissions (5 % of N load) were needed to favor anammox compared with nitratation at low free ammonia (<0.25 mg N?L^?1), low free nitrous acid (<0.9 μg N?L^?1), and higher DO levels (3–4 mg O₂?L^?1). Although the total nitrogen removal rates showed potential, the accumulation of nitrite and nitrate resulted in lower nitrogen removal efficiencies (around 40 %), which should be improved in the future. Moreover, a balance should be found in the future between the increased NO and N₂O emissions and a decreased energy consumption to justify OLAND mainstream treatment.     

施氮对水稻土N2O释放及反硝化功能基因(narG/nosZ)丰度的影响

以紫潮泥和红黄泥两种不同质地的水稻土壤作为研究对象,通过室内培养试验,分析施用硝态氮肥对N2O释放和反硝化基因(narG/nosZ)丰度的影响,并探讨反硝化基因丰度与N2O释放之间的关系。结果表明,施用硝态氮显著增加两种水稻土的N2O释放量。在72h培养过程中,施氮改变了紫潮泥反硝化基因(narG/nosZ)的丰度,但并未明显影响红黄泥反硝化基因(narG/nosZ)丰度。通过双变量相关分析发现,除了紫潮泥narG基因外,其它的反硝化基因丰度和N2O释放之间并没有显著相关性。     

Synthesis, crystal structure, spectroscopic and magnetic properties of copper(II) complexes with 2-(2-pyridyl)imino-N-(2-thiazolin-2-yl)thiazolidine (PyTT)

The copper(II) complexes [Cu(PyTT)₂(H₂O)](NO₃)₂ (A) and [CuCl₂(μ-PyTT)₂CuCl(H₂O)]Cl · 3H₂O (B) were synthesized and characterized by single crystal X-ray diffraction, IR spectroscopy, UV-Vis-NIR diffuse reflectance and magnetic susceptibility measurements. In the mononuclear compound A the copper ion is in a distorted square pyramidal geometry, with the equatorial plane formed by two thiazoline nitrogen atoms, one imino nitrogen atom and one water molecule, whereas the axial site is occupied by one imino nitrogen atom. The compound B is dinuclear and both Cu(II) centres present environments that can be described as slightly distorted square pyramidal geometries. The observed molar magnetic susceptibility for A (μ=2.13 BM) allows to exclude metal-metal interactions, supporting a monomeric structural formulation for this compound. In compound B, magnetic susceptibility measurements in the temperature range 6.2-288 K show an intradimer antiferromagnetic interaction (J=−11.8 cm⁻¹).     

Nitrogen removal in a wastewater treatment plant through biofilters: nitrous oxide emissions during nitrification and denitrification

In order to estimate N₂O emissions from immersed biofilters during nitrogen removal in tertiary treatments at urban wastewater treatment plants (WWTPs), a fixed culture from the WWTP of “Seine Centre” (Paris conurbation) was subjected to lab-scale batch experiments under various conditions of oxygenation and a gradient of methanol addition. The results show that during nitrification, N₂O emissions are positively related to oxygenation (R ² = 0.99). However, compared to the rates of ammonium oxidation, the percentage of emitted N₂O is greater when oxygenation is low (0.5–1 mgO₂ L⁻¹), representing up to 1% of the oxidized ammonium (0.4% on average). During denitrification, the N₂O emission reaches a significant peak when the quantity of methanol allows denitrification of between 66% and 88%. When methanol concentrations lead to a denitrification of close to 100%, the flows of N₂O are much lower and represent on average 0.2% of the reduced nitrate. By considering these results, we can estimate, the emissions of N₂O during nitrogen removal, at the “Seine Centre” WWTP, to approximately 38 kgN-N₂O day⁻¹.     

Performance and optimization of biological nitrogen removal process enhanced by anoxic/oxic step feeding

The aim of this paper is to investigate the performance and optimization of an biological nitrogen removal process enhanced by the anoxic/oxic four stages step-feeding process under sufficient/insufficient influent carbon source. The concept of influent flow distribution ratio (λ) was introduced according to the performance of the step-feed process for inflow splitting and volume distribution. Under the condition of sufficient carbon, the uniform influent flow distribution ratio was proposed from the standpoint of which is easy for process control, operation optimization and management. Under the condition of different influent C/N ratios, the optimal influent flow distribution ratios (λ_opt) were determined, by trial-and-error method. The results showed that more than 95.81% of the total nitrogen removal efficiency could be achieved by step-feeding process with a fixed influent C/N ratio. A model was introduced to derive the relationships of total nitrogen removal efficiency with λ and influent C/N ratio. When the influent carbon source was insufficient, however, the λ_opt between two adjacent stages should not be uniform. The results also showed that the four-stage step-feeding process with λ_opt of 2.0:2.1:2.5:3.4 could significantly improve the total nitrogen removal efficiency. The results obtained in this study will help to optimize the design and the operation of BNR systems.     

Effect of alum on nitrification during simultaneous phosphorous removal in anoxic/oxic reactor

Phosphorus and nitrogen are the important eutrophication nutrients. They are removed in the anoxic/oxic reactor through simultaneous precipitation and biological nitrogen removal. The effect of alum a commonly used simultaneous precipitant on biological nitrification and denitrification are investigated in the present study. Simultaneous removal of phosphorus was carried out using the coagulant alum Al₂(SO₄)₃·14H₂O at 2.2 mol ratio. Before the start of simultaneous precipitation the nitrification rate of the A/O reactor was found to be 0.05 g N-NH₄ ⁺/g VSS/d. It starts to decrease with increase in coagulant dosage. The nitrification rate for alum dosage 97.13 mg/L was 0.38 g N- NH₄ ⁺/g VSS/d. There was no accumulation of nitrate in anoxic tank. The nitrogen removal efficiency of the reactor was affected and it fell from 88 to 78%. There was a slight decrease in effluent COD from 16∼20 mg/L to 8∼12 mg/L after the introduction of simultaneous precipitation into the reactor. The usage of alum as a simultaneous precipitant in the anoxic/oxic reactor was limited due to its inhibition on nitrification. Alum did not have any affect over denitrification process.     

不同施肥对雷竹林径流及渗漏水中氮形态流失的影响

雷竹经营过程中化肥的大量施用,是产区水体污染的主要原因之一,养分管理技术可有效控制面源污染。为了探明减量施肥和有机肥施用对雷竹不同氮形态流失的影响,2012年在浙江省临安市雷竹产区设置了4种施肥处理:对照(CK);常规施肥(CF);减量无机(DI);减量有机无机(DOI),试验于5月18日、9月7日、11月9日分别施用肥料总量的40%,30%和30%,施肥后均进行浅翻,深度5 cm左右。通过建立径流场和土壤渗漏水收集装置,同时在试验田附近布置量雨筒,观察2012年不同氮形态浓度及流失负荷随降雨量的动态变化。研究结果表明:不同施肥处理径流水硝态氮、水溶性有机氮(WSON)以及颗粒态氮的浓度分别在3.82-6.82 mg/L、0.89-1.85 mg/L和0.89-1.83 mg/L,其占总氮的百分比分别为60.9%-68.2%、16.0%-18.1%和15.1%-21.6%。不同施肥处理渗漏水中硝态氮、铵态氮及WSON的浓度分别在26.2-92.5 mg/L、0.50-6.42 mg/L和6.57-12.6 mg/L,其占总氮的百分比分别为75.8%-82.9%、1.50%-6.36%和11.2%-20.6%。不同施肥处理径流水的氮总流失负荷,减量无机和减量有机无机相对于常规施肥来说减少了46.9%和23.1%;不同施肥处理的渗漏水的氮总流失负荷,减量无机和减量有机无机相对于常规施肥来说减少了19.1%和52.1%,可见减量施肥和减量有机无机减少氮流失的效果显著。     

Control of carbon and ammonium ratio for simultaneous nitrification and denitrification in a sequencing batch bioreactor

This study shows how the carbon and nitrogen (C/N) ratio controls the simultaneous occurrence of nitrification and denitrification in a sequencing batch reactor (SBR). Data demonstrated that a low C/N ratio resulted in a rapid carbon deficit, causing an unbalanced simultaneous nitrification–denitrification (SND) process in SBR. When the initial COD/NH₄⁺-N ratio was adjusted to 11.1, the SND-based SBR achieved complete removal of NH₄-N and COD without leaving any NO₂⁻-N in the effluent. The nitrogen removal efficiency decreases gradually with increasing ammonium-loading rate to the SND–SBR system. Altogether, data showed that appropriate controls of carbon and nitrogen input are required to achieve an efficient SND–SBR. An established SND technology can save operation time and energy, and might replace the traditional two-stage biological nitrification and denitrification process.     

Nonlinear response of N2O flux to incremental fertilizer addition in a continuous maize (Zea mays L.) cropping system

The relationship between nitrous oxide (N₂O) flux and N availability in agricultural ecosystems is usually assumed to be linear, with the same proportion of nitrogen lost as N₂O regardless of input level. We conducted a 3‐year, high‐resolution N fertilizer response study in southwest Michigan USA to test the hypothesis that N₂O fluxes increase mainly in response to N additions that exceed crop N needs. We added urea ammonium nitrate or granular urea at nine levels (0–292 kg N ha^?1) to four replicate plots of continuous maize. We measured N₂O fluxes and available soil N biweekly following fertilization and grain yields at the end of the growing season. From 2001 to 2003 N₂O fluxes were moderately low (ca. 20 g N₂O‐N ha^?1 day^?1) at levels of N addition to 101 kg N ha^?1, where grain yields were maximized, after which fluxes more than doubled (to >50 g N₂O‐N ha^?1 day^?1). This threshold N₂O response to N fertilization suggests that agricultural N₂O fluxes could be reduced with no or little yield penalty by reducing N fertilizer inputs to levels that just satisfy crop needs.     

Nitrogen removal and microbial communities in a three-stage system simulating a riparian environment

The riparian zone is an active interface for nitrogen removal, in which nitrogen transformations by microorganisms have not been valued. In this study, a three-stage system was constructed to simulate the riparian zone environments, and nitrogen removal as well as the microbial community was investigated in this ‘engineered riparian system’. The results demonstrated that stage 1 of this system accounted for 41–51 % of total nitrogen removal. Initial ammonium loading and redox potential significantly impacted the nitrogen removal performances. Stages 1 and 2 were both composed of an anoxic/oxic (A/O) zone and an anaerobic column. The A/O zone removed most of the ammonium load (6.8 g/m²/day), while the anaerobic column showed a significant nitrate removal rate (11.1 g/m²/day). Molecular biological analysis demonstrated that bacterial diversity was high in the A/O zones, where ammonium-oxidizing bacteria and nitrite-oxidizing bacteria accounted for 8.42 and 3.32 % of the bacterial population, respectively. The denitrifying bacteria Acidovorax sp. and the nitrifying bacteria Nitrosospira/Nitrosomonas were the predominant microorganisms in this engineered riparian system. This three-stage system was established to achieve favorable nitrogen removal and the microbial community in the system was also retained. This investigation should deepen our understanding of biological nitrogen removal in engineered riparian zones.     

Prediction and inhibition of the N<Subscript>2</Subscript>O accumulation in the BioDeNO<Subscript><Emphasis Type="Italic">x</Emphasis></Subscript> process for NO<Subscript><Emphasis Type="Italic">x</Emphasis></Subscript> removal from flue gas

BioDeNO _x process, which combines the advantages of the chemical absorption and biological reduction processes, is regarded as a promising candidate for NO removal from the flue gas. In the BioDeNO _x , N₂O was accumulated in the process of the biological reduction of Fe^II(EDTA)-NO. In this work, the pathway of the Fe^II(EDTA)-NO reduction was investigated and a mathematic model was developed to evaluate and predict the accumulation of N₂O. Furthermore, parametric tests such as the effects of the C/N ratio (molar ratio of carbon/nitrogen), electron donor, and sulfite concentrations on N₂O accumulation were investigated. Experimental results revealed that N₂O accumulation was inhibited with a high C/N ratio (2.4), sufficient electron donor, and a low sulfite concentration. In addition, compared with the inorganic electron donor (Fe^II(EDTA)), the organic electron donor (glucose) was beneficial for microorganism metabolism and N₂O accumulation inhibition. This work will provide significant insight into the inhibition of N₂O accumulation during the operation of BioDeNO _x and advance this novel process for the industrial application.