Physical mixing in coastal waters controls and decouples nitrification via biomass dilution.

Nitrification is a central process of the aquatic nitrogen cycle that controls the supply of nitrate used in other key processes, such as phytoplankton growth and denitrification. Through time series observation and modeling of a seasonally stratified, eutrophic coastal basin, we demonstrate that physical dilution of nitrifying microorganisms by water column mixing can delay and decouple nitrification. The findings are based on a 4-y, weekly time series in the subsurface water of Bedford Basin, Nova Scotia, Canada, that included measurement of functional (amoA) and phylogenetic (16S rRNA) marker genes. In years with colder winters, more intense winter mixing resulted in strong dilution of resident nitrifiers in subsurface water, delaying nitrification for weeks to months despite availability of ammonium and oxygen. Delayed regrowth of nitrifiers also led to transient accumulation of nitrite (3 to 8 µmol · kgsw-1) due to decoupling of ammonia and nitrite oxidation. Nitrite accumulation was enhanced by ammonia-oxidizing bacteria (Nitrosomonadaceae) with fast enzyme kinetics, which temporarily outcompeted the ammonia-oxidizing archaea (Nitrosopumilus) that dominated under more stable conditions. The study reveals how physical mixing can drive seasonal and interannual variations in nitrification through control of microbial biomass and diversity. Variable, mixing-induced effects on functionally specialized microbial communities are likely relevant to biogeochemical transformation rates in other seasonally stratified water columns. The detailed study reveals a complex mechanism through which weather and climate variability impacts nitrogen speciation, with implications for coastal ecosystem productivity. It also emphasizes the value of high-frequency, multiparameter time series for identifying complex controls of biogeochemical processes in aquatic systems.

Sudden sulfamethoxazole shock leads to nitrite accumulation in microalgae-nitrifying bacteria consortia: Physiological responses and light regulating strategy.

Antibiotic shock may potentially impact the performance of promising microalgae-nitrifying bacteria consortia (MNBC) processes. This study investigated physiological behaviors of MNBC under sulfamethoxazole (SMX) shock (mg/L level) and verified a light regulating strategy for improving process performance. Results showed that SMX shock did not affect ammonium removal but caused nitrite accumulation, resulting from combined effects of excessive reactive oxidative species (ROS) production, inhibited microalgal photosynthetic activity, upregulated expressions of amoA and hao, and downregulated expression of nxrA. Moreover, high ammonium concentration aggravated nitrite accumulation and reduced ammonium removal owing to significantly reduced dissolved oxygen (DO). Increasing light intensity enhanced microalgal photo-oxygenation and promoted expressions of all nitrification-related genes, thus improving ammonium removal and alleviating nitrite accumulation. A central composite design coupled with response surface methodology (CCD-RSM) further demonstrated the negative impacts of SMX shock and high ammonium on MNBC and the effectiveness of the light regulation in maintaining stable process performance. This study provides theoretical basis for physiological responses and regulatory strategy of the MNBC process facing short-term antibiotic shock.

Optimal conditions and nitrogen removal performance of aerobic denitrifier Comamonas sp. pw-6 and its bioaugmented application in synthetic domestic wastewater treatment.

To assess the possibility of using aerobic denitrification (AD) bacteria with high NO2--N accumulation for nitrogen removal in wastewater treatment, conditional optimization, as well as sole and mixed nitrogen source tests involving AD bacterium, Comamonas sp. pw-6 was performed. The results showed that the optimal carbon source, pH, C/N ratio, rotational speed, and salinity for this strain were determined to be succinate, 7, 20, 160 rpm, and 0%, respectively. Further, this strain preferentially utilized NH4+-N, NO3--N, and NO2--N, and when NO3--N was its sole nitrogen source, 92.28% of the NO3--N (150 mg·L-1) was converted to NO2--N. However, when NH4+-N and NO3--N constituted the mixed nitrogen source, NO3--N utilization by this strain was significantly lower (p < 0.05). Therefore, a strategy was proposed to combine pw-6 bacteria with traditional autotrophic nitrification to achieve the application of pw-6 bacteria in NH4+-N-containing wastewater treatment. Bioaugmented application experiments showed significantly higher NH4+-N removal (5.96 ± 0.94 mg·L-1·h-1) and lower NO3--N accumulation (2.52 ± 0.18 mg·L-1·h-1) rates (p < 0.05) than those observed for the control test. Thus, AD bacteria with high NO2--N accumulation can also be used for practical applications, providing a basis for expanding the selection range of AD strains for wastewater treatment.

Deregulation of Ribosome Biogenesis in Nitrite-Oxidizing Bacteria Leads to Nitrite Accumulation.

Nitrite (NO2-) accumulation caused by nitrite-oxidizing bacteria (NOB) inhibition in nitrification is a double-edged sword, i.e., a disaster in aquatic environments but a hope for innovating nitrogen removal technology in wastewater treatment. However, little information is available regarding the molecular mechanism of NOB inhibition at the cellular level. Herein, we investigate the response of NOB inhibition on NO2- accumulation established by a side-stream free ammonia treatment unit in a nitrifying reactor using integrated metagenomics and metaproteomics. Results showed that compared with the baseline, the relative abundance and activity of NOB in the experimental stage decreased by 91.64 and 68.66%, respectively, directly resulting in a NO2- accumulation rate of 88%. Moreover, RNA polymerase, translation factors, and aa-tRNA ligase were significantly downregulated, indicating that protein synthesis in NOB was interfered during NO2- accumulation. Further investigations showed that ribosomal proteins and GTPases, responsible for bindings between either ribosomal proteins and rRNA or ribosome subunits, were remarkably downregulated. This suggests that ribosome biogenesis was severely disrupted, which might be the key reason for the inhibited protein synthesis. Our findings fill a knowledge gap regarding the underlying mechanisms of NO2- accumulation, which would be beneficial for regulating the accumulation of NO2- in aquatic environments and engineered systems.

Nitrite accumulation in activated sludge through cyclic anaerobic exposure with acetate.

Achieving nitrite accumulation still remains challenging for efficient short-cut biological nitrogen removal in municipal wastewater treatment. To tackle the problem of insufficient carbon in incoming wastewater for biological nutrient removal, a return activated sludge (RAS) fermentation method has been proposed and demonstrated to enable producing supplemental volatile fatty acids (VFAs) and enhance biological phosphorus removal via sludge cycling between mainstream and a sidestream anaerobic reactor. However, the impacts of long anaerobic exposure with acetate on nitrifying bacteria, known as the aerobic chemoautotrophic microorganisms, remains unexplored. In this study, the activated sludge underwent a cyclic anaerobic treatment with the addition of acetate (Ac), the effects on nitrification rate, abundance and microdiversity of nitrifying communities were comprehensively assessed. Firstly, batch activity tests proved the direct addition of high acetate (above 1000 mg/L) could cause inhibition on the nitrification rate, moreover, the inhibitory effect was stronger on nitrite-oxidizing bacteria (NOB) activity than that of ammonia-oxidizing bacteria (AOB). Then, a sequencing batch reactor (SBR) was applied to test the nitrogen conversion performance for low-strength ammonium wastewater. Nitrite accumulation could be achieved via the cyclic anaerobic exposure with 1000-5000 mg Ac/L. The maximum effluent concentration of nitrite was 40.8 ± 3.5 mg N/L with nitrite accumulation ratio (NAR) of 67.6 ± 3.5%. The decrease in NOB activity (72.7%) was greater than AOB of 42.4%, promoting nitrite accumulation via nitritation process. Furthermore, the cyclic anaerobic exposure with acetate can largely reshape the nitrifying communities. As the dominant AOB and NOB, the abundance of Nitrosomonas and Nitrospira were both decreased with species-level microdiversity in the nitrifying communities. However, the heterotrophic microorganism, Thauera, were found to be highly enriched (from 0 to 17.3%), which may act as the potential nitrite producer as proved by the increased nitrate reduction gene abundance. This study can provide new insights into achieving mainstream nitrite accumulation by involving sidestream RAS fermentation towards efficient wastewater treatment management.

Mitigating nitrite accumulation during S0-based autotrophic denitrification: Balancing nitrate-nitrite reduction rate with thiosulfate as external electron donor.

This study was carried out to determine the effect of influent nitrate loading on nitrite accumulation during elemental-sulfur based denitrification process, and proposed to enhance the nitrogen removal efficiency by mitigating nitrite accumulation with thiosulfate as external electron donor. Along with increasing the nitrate influent loading (from 0.09 kg N/m3/d to 1.73 kg N/m3/d) by shortening the empty bed contact time (EBCT) (from 5 h to 0.25 h), the nitrate removal loading increased from 0.08 to 0.83 kg N/m3/d. Meanwhile, the raise of the nitrate influent loading obviously aggravated the nitrite accumulation. Herein, nitrite began to accumulate since the nitrate influent loading was over 0.86 kg N/m3/d, and a maximum nitrite accumulation of 2.39 mg/L was observed under the 0.25 h of EBCT and 15 mg/L of nitrate influent concentration condition. Thiosulfate was used as the external electron donor to accelerate the nitrite reduction rate in order to mitigate the nitrite accumulation. As a result, the nitrite accumulation significantly decreased from 2.39 mg/L to 0.17 mg/L with the thiosulfate dosage of 13.36 mg/L. However, the nitrite accumulation bounced with the on-going increase of the thiosulfate dosage, indicating that the nitrate reduction rate and nitrite reduction rate were accelerated alternatively. After dosing thiosulfate, the relative abundances of sulfurimonas and ferritrophicum grew up significantly.

Bioaugmentation for low C/N ratio wastewater treatment by combining endogenous partial denitrification (EPD) and denitrifying phosphorous removal (DPR) in the continuous A2/O - MBBR system.

Endogenous partial denitrification (EPD) and denitrifying phosphorous removal (DPR) were combined in a novel A2/O - MBBR (Anaerobic Anoxic Oxic - Moving Bed Biofilm Reactor) system for low carbon/nitrogen (C/N) ratio wastewater treatment. The DPR performance was compared and the nutrient metabolism was elucidated based on the optimization of hydraulic retention time (HRT, 4-12 h) and nitrate recycling (R, 200%-600%). In the continuous-flow, the nitrate (NO3-) denitrification accompanied by nitrite (NO2-, via EPD) accumulation with the nitrate-to-nitrite transformation ratio (NTR) of 35.87%-43.31% in the anoxic zones. At HRT of 12 h with R of 500%, batch test initially revealed the DPR mechanism using both NO3- and NO2- as electron acceptor, where denitrifying phosphorus accumulation organisms (DPAOs) and denitrifying glycogen accumulation organisms (DGAOs) were the main contributors for EPD with incomplete denitrification (NO3- → NO2-). Furthermore, stoichiometry-based functional bacteria analysis displayed that higher bioactivity of DPAOs (NO2-→N2, 57.30%; NO3-→N2, 35.85%) over DGAOs (NO3-→N2, 6.85%) facilitated the anoxic NO3- reduction. Microbial community analysis suggested that Cluster I of Defluviicoccus-GAO group (∼4%) was responsible for stable NO2- accumulation performance via EPD, while increased Accumulibacter-PAO group (by ∼15%) contributed to the advanced nutrient removal. Based on the achievement of NO2- accumulation, the application feasibility of integrated EPD - DPR - Anammox for deep-level nutrient removal was discussed.

Application of humin-immobilized biocathode in a continuous-flow bioelectrochemical system for nitrate removal at low temperature.

Solid-phase humic substances (humin) can work as an additional electron donor to support the low temperature denitrification but the reducing capacity of its non-reduced form is limited. In this study, a continuous-flow denitrifying BES with a humin-immobilized biocathode (H-BioC) was established. Humin was expected to function as a redox mediator and be persistently reduced on the cathode to provide reducing power to a denitrifying biofilm. Results showed that the H-BioC maintained a stable denitrification capacity with low nitrite accumulation for more than 100 days at 5 °C, and the specific microbial denitrification rate and electron transfer rate were 3.97-fold and 1.75-fold higher than those of the unaltered cathode. The results of repeated cycles of humin reduction and oxidation experiments further suggested that the redox activity of humin was stable. Acidovorax was the most dominant genus in both H-BioC biofilm and unaltered cathodic biofilm, while Rhodocyclaceae (unclassified_f_) was more enriched in H-BioC biofilm. Further Phylogenetic Investigation of Communities by Reconstruction of Unobserved States (PICRUSt) analyses indicated that biofilm formation, electron transfer, and nitrate reduction functions were more abundant in H-BioC, suggesting a possible enhancement mechanism by humin. The results of this study raise the possibility that immobilization of solid-phase humin may be a useful strategy for electrostimulated heterotrophic denitrification in groundwater where the indigenous bacteria have poor electroactivity.

A predictive model for N2O production in anammox-granular sludge reactors: Combined effects of nitrite/ammonium ratio and organic matter concentration.

Once the use of anammox reactors has been increasing on a global scale, it is important to understand the mechanisms of N2O emissions and how to minimise the emissions by optimising the operating conditions. In this study, the influence of chemical oxygen demand (COD) (from 0 mgO2 L-1 to 100 mgO2 L-1) and nitrite/ammonium ratio from 0.79 to 2.21 (maintaining ammonium at 100 mgN L-1 and varying nitrite from 79 mgN L-1 to 221 mgN L-1) in the N2O emissions from anammox-granular sludge reactor was investigated in two steps. Step 1 consisted of batch tests, using central composite design, and Step 2, long-term operation of a 6.5 L continuous up-flow reactor. The results showed that the N2O emissions were minimized by controlling, in the influent, the NO2--N/NH4+-N ratio from 1.1 to 1.3 and maintaining the COD concentration below 100 mgO2 L-1. TN removal efficiencies were higher than 70% in all conditions tested".

Single-Atom Cu Catalysts for Enhanced Electrocatalytic Nitrate Reduction with Significant Alleviation of Nitrite Production.

Metallic Cu is a well-known electrocatalyst for nitrate reduction reaction (NO3 RR), but it suffers from relatively low activity, poor stability, and inducing nitrite accumulation during the long-term operation. Herein, it is found that Cu catalysts minimized at the single-atom level can overcome the limitations of bulk materials in NO3 RR. A metal-nitrogen-carbon (M-N-C) electrocatalyst composed of carbon nanosheets embedding isolated copper atoms coordinated with N, Cu-N-C-800, is synthesized by pyrolysis of a Cu-based metal-organic framework at 800 °C. In comparison with Cu nanoparticles and Cu plate-800, kinetic measurements show that the Cu-N-C-800 electrocatalyst is more active and stable and distinctly suppresses the release of nitrite intermediate into the solution. The combined results of experimental data and density functional theory calculations indicate that Cu bound with N (particularly Cu-N2 ) is the key to favorable adsorption of NO3 - and NO2 - . This strong binding is responsible for the enhanced rate of nitrate conversion to the end products of ammonia and nitrogen. These findings highlight the promise of single-atom Cu electrocatalysts for nitrate reduction with desirable performance.

New direction in biological nitrogen removal from industrial nitrate wastewater via anammox.

Anaerobic ammonium oxidation (anammox) is an important scientific discovery in the field of wastewater treatment. This process is a sustainable option in nitrogen removal due to its energy-efficient and cost-effective advantage. Great effort has been made recently to remove ammonium from industrial and municipal wastewater via the anammox process with a preceding partial nitrification (PN) converting part of NH4+ to NO2-. Anammox process is seldom involved in the nitrate removal. Nitrate (NO3-), one of the main nitrogen compounds produced from various industries, is typically converted to nitrogen gas via denitrification process where a large amount of carbon source is consumed. Within this context, we reviewed the current technologies for high-strength nitrate wastewater treatment. It is found that nitrite accumulation often occurs during nitrate reduction, and its accumulating level would be increased at certain conditions (i.e., low C/N ratio and high pH). Hence, this provides a great opportunity to employ the anammox process to further convert nitrite in a more sustainable way. In this review, we highlight a new approach for industrial nitrate wastewater treatment via partial denitrification coupled with anammox process (PD-A). We also discuss the conditions to achieve successful PD-A process, economic and environmental benefits, and potential challenges as well as the future perspectives in practical application.

Influence of Hydrogen Electron Donor, Alkaline pH, and High Nitrate Concentrations on Microbial Denitrification: A Review.

Bacterial respiration of nitrate is a natural process of nitrate reduction, which has been industrialized to treat anthropic nitrate pollution. This process, also known as "microbial denitrification", is widely documented from the fundamental and engineering points of view for the enhancement of the removal of nitrate in wastewater. For this purpose, experiments are generally conducted with heterotrophic microbial metabolism, neutral pH and moderate nitrate concentrations (<50 mM). The present review focuses on a different approach as it aims to understand the effects of hydrogenotrophy, alkaline pH and high nitrate concentration on microbial denitrification. Hydrogen has a high energy content but its low solubility, 0.74 mM (1 atm, 30 °C), in aqueous medium limits its bioavailability, putting it at a kinetic disadvantage compared to more soluble organic compounds. For most bacteria, the optimal pH varies between 7.5 and 9.5. Outside this range, denitrification is slowed down and nitrite (NO2-) accumulates. Some alkaliphilic bacteria are able to express denitrifying activity at pH levels close to 12 thanks to specific adaptation and resistance mechanisms detailed in this manuscript, and some bacterial populations support nitrate concentrations in the range of several hundred mM to 1 M. A high concentration of nitrate generally leads to an accumulation of nitrite. Nitrite accumulation can inhibit bacterial activity and may be a cause of cell death.

Effects of C/N ratio on nitrate removal and floc morphology of autohydrogenotrophic bacteria in a nitrate-containing wastewater treatment process.

The effects of C/N ratio of a nitrate-containing wastewater on nitrate removal performed by autohydrogenotrophic bacteria as well as on the morphological parameters of floc such as floc morphology, floc number distribution, mean particle size (MPS), aspect ratio and transparency were examined in this study. The results showed that the nitrate reduction rate increased with increasing C/N ratio from 0.5 to 10 and that the nitrogen removal of up to 95% was found at the C/N ratios of higher than 5 (between 0.5-10). Besides, high C/N ratio values reflected a corresponding high nitrite accumulation after 12-hr operation, and a fast decreasing rate of nitrite in the rest of operational time. The final pH values increased with the C/N ratio increasing from 0.5 to 2.5, but decreased with the C/N ratio increasing from 2.5 to 10. There were no significant changes in floc morphology with the MPSs ranging from 35 to 40µm. Small and medium-sized flocs were dominant in the sludge suspension, and the number of flocs increased with the increasing C/N ratios. Furthermore, the highest apparent frequency of 10% was observed at aspect ratios of 0.5 and 0.6, while the transparency of flocs changed from 0.1 to 0.7.

Comparison of oxygen transfer parameters and oxygen demands in bioreactors operated at low and high dissolved oxygen levels.

The proper design of aeration systems for bioreactors is critical since it can represent up to 50% of the operational and capital cost at water reclamation facilities. Transferring the actual amount of oxygen needed to meet the oxygen demand of the wastewater requires α- and ß-factors, which are used for calculating the actual oxygen transfer rate (AOTR) under process conditions based on the standard oxygen transfer rate (SOTR). The SOTR is measured in tap water at 20°C, 1 atmospheric pressure, and 0 mg L-1 of dissolved oxygen (DO). In this investigation, two 11.4-L bench-scale completely mixed activated process (CMAS) reactors were operated at various solid retention times (SRTs) to ascertain the relationship between the α-factor and SRT, and between the ß-factor and SRT. The second goal was to determine if actual oxygen uptake rates (AOURs) are equal to calculated oxygen uptake rates (COURs) based on mass balances. Each reactor was supplied with 0.84 L m-1 of air resulting in SOTRs of 14.3 and 11.5 g O2 d-1 for Reactor 1 (R-1) and Reactor 2 (R-2), respectively. The estimated theoretical oxygen demands of the synthetic feed to R-1 and R-2 were 6.3 and 21.9 g O2 d-1, respectively. R-2 was primarily operated under a dissolved oxygen (DO) limitation and high nitrogen loading to determine if nitrification would be inhibited from a nitrite buildup and if this would impact the α-factor. Nitrite accumulated in R-2 at DO concentrations ranging from 0.50 to 7.35 mg L-1 and at free ammonia (FA) concentrations ranging from 1.34 to 7.19 mg L-1. Nonsteady-state reaeration tests performed on the effluent from each reactor and on tap water indicated that the α-factor increased as SRT increased. A simple statistical analysis (paired t-test) between AOURs and COURs indicated that there was a statistically significant difference at 0.05 level of significance for both reactors. The ex situ BOD bottle method for estimating AOUR appears to be invalid in bioreactors operated at low DO concentrations (<1.0 mg L-1).

Enrichment of denitratating bacteria from a methylotrophic denitrifying culture.

Denitratation (nitrite produced from nitrate), has the potential applications in wastewater treatment by combining with ANAMMOX process. The occurrence of denitratation has been shown to be effected qualitatively by various parameters in the environment. A more quantitative understanding can be obtained using enrichment cultures in lab-scale experiments, yet information on the enrichment of functional microorganisms responsible for denitratation is lacking. In this study, a stable denitratation-dominated culture was obtained from methylotrophic denitrifying culture. The results showed that, besides the substitution of acetate for methanol, the lasting starvation following saturation of electron donor was another pivotal selection pressure that favored the growth of denitratating bacteria, which was supported by the distinctive physiological strategy involving the higher growth rate combining with larger poly-hydroxybutyrate (PHB) accumulation at sufficient electron donor situation and then manage the stress of electron donor starvation by consumpiton of the PHB. High-throughput 16S rRNA gene sequencing analysis indicated that non-methylotrophic Halomonas campisalis (48.1 %) and Halomonas campaniensis (30.4 %) dominated in the denitratating community. Moreover the denitratation was driven by the nitrate inhibiting the nirS transcription in the Halomonas species.

Response of nitrite accumulation and microbial community to free ammonia and dissolved oxygen treatment of high ammonium wastewater.

The effects of free ammonia (FA) and dissolved oxygen (DO) on nitrite accumulation in the treatment of high ammonium wastewater and on the evolution of the microbial community were investigated. Under high DO conditions (3.75 ± 0.49 mg/L), FA as high as 10.61 ± 2.89 mg NH3/L maintained stable nitrite accumulation rate (NAR) of 84 % with NH4 (+)-N load of 2.05 kg N/(m(3) day) at sludge retention time (SRT) of 15-18 days. After 56 days of operation, Proteobacteria and Nitrosomonas were the dominant phylum and genus, respectively; Nitrosomonas increased from 21.14 to 54.57 %. By contrast, under relative low DO and low FA, nitrite-oxidizing bacteria (NOB) were nearly eliminated (NOB/AOB of 0; ammonium-oxidizing bacteria (AOB)), and NAR of 94 % was achieved with lower NH4 (+)-N load of 0.48 kg N/(m(3) day). DO correlated with AOB and NOB abundance, and FA decreased NOB activity and the NOB/AOB ratio. In conclusion, high FA and high DO conditions are optimal for efficient nitrite accumulation.

Mechanisms and microbial structure of partial denitrification with high nitrite accumulation.

Nitrite (NO2 (-)-N) accumulation in denitrification can provide the substrate for anammox, an efficient and cost-saving process for nitrogen removal from wastewater. This batch-mode study aimed at achieving high NO2 (-)-N accumulation over long-term operation with the acetate as sole organic carbon source and elucidating the mechanisms of NO2 (-)-N accumulation. The results showed that the specific nitrate (NO3 (-)-N) reduction rate (59.61 mg N VSS(-1) h(-1) at NO3 (-)-N of 20 mg/L) was much higher than specific NO2 (-)-N reduction rate (7.30 mg N VSS(-1) h(-1) at NO3 (-)-N of 20 mg/L), and the NO2 (-)-N accumulation proceeded well at the NO3 (-)-N to NO2 (-)-N transformation ratio (NTR) as high as 90 %. NO2 (-)-N accumulation was barely affected by the ratio of chemical oxygen demand (COD) to NO3 (-)-N concentration (C/N). With the addition of NO3 (-)-N, NO2 (-)-N accumulation occurred and the specific NO2 (-)-N reduction rate declined to a much lower level compared with the value in the absence of NO3 (-)-N. This indicated that the denitrifying bacteria in the system preferred to use NO3 (-)-N as electron acceptor rather than use NO2 (-)-N. In addition, the Illumina high-throughput sequencing analysis revealed that the genus of Thauera bacteria was dominant in the denitrifying community with high NO2 (-)-N accumulation and account for 67.25 % of total microorganism. This bacterium might be functional for high NO2 (-)-N accumulation in the presence of NO3 (-)-N.

Aerobic N2O emission for activated sludge acclimated under different aeration rates in the multiple anoxic and aerobic process.

Nitrous oxide (N2O) is a potent greenhouse gas that can be emitted during biological nitrogen removal. N2O emission was examined in a multiple anoxic and aerobic process at the aeration rates of 600mL/min sequencing batch reactor (SBRL) and 1200mL/min (SBRH). The nitrogen removal percentage was 89% in SBRL and 71% in SBRH, respectively. N2O emission mainly occurred during the aerobic phase, and the N2O emission factor was 10.1% in SBRL and 2.3% in SBRH, respectively. In all batch experiments, the N2O emission potential was high in SBRL compared with SBRH. In SBRL, with increasing aeration rates, the N2O emission factor decreased during nitrification, while it increased during denitrification and simultaneous nitrification and denitrification (SND). By contrast, in SBRH the N2O emission factor during nitrification, denitrification and SND was relatively low and changed little with increasing aeration rates. The microbial competition affected the N2O emission during biological nitrogen removal.

Nitrite accumulation from simultaneous free-ammonia and free-nitrous-acid inhibition and oxygen limitation in a continuous-flow biofilm reactor.

To achieve nitrite accumulation for shortcut biological nitrogen removal (SBNR) in a biofilm process, we explored the simultaneous effects of oxygen limitation and free ammonia (FA) and free nitrous acid (FNA) inhibition in the nitrifying biofilm. We used the multi-species nitrifying biofilm model (MSNBM) to identify conditions that should or should not lead to nitrite accumulation, and evaluated the effectiveness of those conditions with experiments in continuous flow biofilm reactors (CFBRs). CFBR experiments were organized into four sets with these expected outcomes based on the MSNBM as follows: (i) Control, giving full nitrification; (ii) oxygen limitation, giving modest long-term nitrite build up; (iii) FA inhibition, giving no long-term nitrite accumulation; and (iv) FA inhibition plus oxygen limitation, giving major long-term nitrite accumulation. Consistent with MSNBM predictions, the experimental results showed that nitrite accumulated in sets 2-4 in the short term, but long-term nitrite accumulation was maintained only in sets 2 and 4, which involved oxygen limitation. Furthermore, nitrite accumulation was substantially greater in set 4, which also included FA inhibition. However, FA inhibition (and accompanying FNA inhibition) alone in set 3 did not maintained long-term nitrite accumulation. Nitrite-oxidizing bacteria (NOB) activity batch tests confirmed that little NOB or only a small fraction of NOB were present in the biofilms for sets 4 and 2, respectively. The experimental data supported the previous modeling results that nitrite accumulation could be achieved with a lower ammonium concentration than had been required for a suspended-growth process. Additional findings were that the biofilm exposed to low dissolved oxygen (DO) limitation and FA inhibition was substantially denser and probably had a lower detachment rate.

N2O emission from nitrogen removal via nitrite in oxic-anoxic granular sludge sequencing batch reactor.

Bionitrification is considered to be a potential source of nitrous oxide (N2O) emissions, which are produced as a by-product during the nitrogen removal process. To investigate the production of N2O during the process of nitrogen removal via nitrite, a granular sludge was studied using a lab-scale sequence batch reactor operated with real-time control. The total production of N2O generated during the nitrification and denitrification processes were 1.724 mg/L and 0.125 mg/L, respectively, demonstrating that N2O is produced during both processes, with the nitrification phase generating larger amount. In addition, due to the N2O-N mass/oxidized ammonia mass ratio, it can be concluded that nitrite accumulation has a positive influence on N2O emissions. Results obtained from PCR-DGGE analysis demonstrate that a specific Nitrosomonas microorganism is related to N2O emission.