Nitrous oxide production in autotrophic nitrogen removal granular sludge: A modeling study

The sustainability of autotrophic granular system performing partial nitritation and anaerobic ammonium oxidation (anammox) for complete nitrogen removal is impaired by the production of nitrous oxide (N₂O). A systematic analysis of the pathways and affecting parameters is, therefore, required for developing N ₂O mitigation strategies. To this end, a mathematical model capable of describing different N ₂O production pathways was defined in this study by synthesizing relevant mechanisms of ammonium-oxidizing bacteria (AOB), nitrite-oxidizing bacteria, heterotrophic bacteria (HB), and anammox bacteria. With the model validity reliably tested and verified using two independent sets of experimental data from two different autotrophic nitrogen removal biofilm/granular systems, the defined model was applied to reveal the underlying mechanisms of N ₂O production in the granular structure as well as the impacts of operating conditions on N ₂O production. The results show that: (a) in the aerobic zone close to the granule surface where AOB contribute to N ₂O production through both the AOB denitrification pathway and the NH ₂OH pathway, the co-occurring HB consume N ₂O produced by AOB but indirectly enhance the N ₂O production by providing NO from NO ₂ ⁻ reduction for the NH ₂OH pathway, (b) the inner anoxic zone of granules with the dominance of anammox bacteria acts as a sink for NO ₂ ⁻ diffusing from the outer aerobic zone and, therefore, reduces N ₂O production from the AOB denitrification pathway, (c) operating parameters including bulk DO, influent NH ₄ ⁺, and granule size affect the N ₂O production in the granules mainly by regulating the NH ₂OH pathway of AOB, accounting for 34–58% of N ₂O turnover, and (d) the competition between the NH ₂OH pathway and heterotrophic denitrification for nitric oxide leads to the positive role of HB in reducing N ₂O production in the autotrophic nitrogen removal granules, which could be further enhanced in the presence of a proper level of influent organics.     

Nitrification and Denitrification in High-Strength Ammonium by <Emphasis Type="Italic">Alcaligenes faecalis</Emphasis>

Alcaligenes faecalis sp. No. 4, that has the ability of heterotrophic nitrification and aerobic denitrification in high-strength ammonium at about 1200 mg-N/l, converted about one-half of removed NH ₄⁺-N to intracellular nitrogen and nitrified only 3% of the removed NH₄⁺. From the nitrogen balance, 40–50% of removed NH₄⁺-N was estimated to be denitrified. Production of N₂ was confirmed by GC-MS and 90% of denitrified products was N₂. The maximum ammonium removal rate, 29 mg-N/l h and its denitrification rate in aerated batch experiments, were 5–40 times higher than those of other bacteria with the same ability.     

In situ simultaneous organics and nitrogen removal from recycled landfill leachate using an anaerobic-aerobic process

An anaerobic-aerobic process including a fresh refuse landfill reactor as denitrifying reactor, a well-decomposed refuse reactor as methanogenesis reactor and an aerobic activated sludge reactor as nitrifying reactor was operated by leachate recirculation to remove organic and nitrogen simultaneously. The results indicated that denitrification and methanogenesis were carried out successfully in the fresh refuse and well-decomposed landfill reactors, respectively, while the nitrification of NH(4)(+)-N was performed in the aerobic reactor. The maximum organic removal rate was 1.78 kg COD/m(3)d in the well-decomposed refuse landfill reactor while the NH(4)(+)-N removal rate was 0.18 kg NH(4)(+)-N/m(3)d in the aerobic reactor. The biogas from fresh refuse reactors and well-decomposed refuse landfill reactors were consisted of mainly carbon dioxide and methane, respectively. The volume fraction of N(2) increased with the increase of NO(3)(-)-N concentration and decreased with the drop of NO(3)(-)-N concentration. The denitrifying bacteria mustered mainly in middle layer and the denitrifying bacteria population had a good correlation with NO(3)(-)-N concentration.     

Removal of nitrogen by heterotrophic nitrification–aerobic denitrification of a novel halotolerant bacterium Pseudomonas mendocina TJPU04

Excess inorganic nitrogen in water poses a severe threat to enviroment. Removal of inorganic nitrogen by heterotrophic nitrifying–aerobic denitrifying microorganism is supposed to be a promising and applicable technology only if the removal rate can be maintained sufficiently high in real wastewater under various conditions, such as high concentration of salt and wide range of different nitrogen concentrations. Here, a new heterotrophic nitrifying–aerobic denitrifying bacterium was isolated and named as Pseudomonas mendocina TJPU04, which removes NH₄⁺-N, NO₃⁻-N and NO₂⁻-N with average rate of 4.69, 5.60, 4.99 mg/L/h, respectively. It also maintains high nitrogen removal efficiency over a wide range of nitrogen concentrations. When concentration of NH₄⁺-N, NO₃⁻-N and NO₂⁻-N was up to 150, 150 and 50 mg/L, 98%, 93%, and 100% removal efficiency could be obtained, respectively, after 30-h incubation under sterile condition. When it was applied under non-sterile condition, the ammonia removal efficiency was slightly lower than that under sterile condition. However, the nitrate and nitrite removal efficiencies under non-sterile condition were significantly higher than those under sterile condition. Strain TJPU04 also showed efficient nitrogen removal performance in the presence of high concentration of salt and nitrogen. In addition, the removal efficiencies of NH₄⁺-N, NO₃⁻-N and TN in real wastewater were 91%, 52%, and 75%, respectively. These results suggest that strain TJPU04 is a promising candidate for efficient removal of inorganic nitrogen in wastewater treatment.

     

Wastewater nitrification kinetics using reciprocating jet bioreactor

Kinetic investigations on growth parameters of nitrifying and COD oxidizing bacteria were carried out with recourse to a three stage reciprocating jet bioreactor system using real life wastewater. The system employed in this investigation essentially consisted of separate aerobic oxidation stage along with nitrification stage and anaerobic denitrification stage with facility for biomass recirculation whenever necessary. Steady-state COD oxidation reactor performance was assessed for various values of residence time. Yield coefficient and decay coefficient of COD oxidizing biomass were obtained as 0.3329 kg BM/kg COD and 0.0032 (1/h) respectively.It was observed that COD oxidizing bacteria co-existed with nitrifying bacteria during nitrification process due to the nature of wastewater used. Steady-state nitrification reactor performance was also assessed for various residence time values. Exact concentration of nitrifying and COD oxidizing biomass in the nitrification reactor was then estimated with the help of kinetic growth parameters of COD oxidizing biomass and extent of COD oxidation achieved in nitrification reactor. This further enabled evaluation of corrected kinetic growth parameters estimated as 0.4272 kg BM/kg NH ₄ ⁺ -N and 0.00626 (1/h) for nitrifier biomass yield coefficient and decay coefficient respectively.     

Nitrogen mineralization in acid sulfate soils

Summary Mineralization of soil nitrogen studies with two acid sulfate soils under anaerobic and aerobic incubation at 30°C for 2 weeks showed that the mineral N was released and accumulated entirely as NH ₄ ⁺ in both soils. Nitrification did not occur in either of the soils under conditions that stimulate nitrification. The acid sulfate soils studied release good amounts of mineralizable N, and, because of lack of nitrifying activity, denitrification may not be a serious problem in these soils.     

Gaseous nitrogen losses and ammonia volatilization measurement following land application of cattle slurry in the mid-Atlantic region of the USA

To provide locally-determined field data for extension and environmental management purposes, gaseous N losses were measured following cattle slurry application to an arable silty-loam soil in the mid-Atlantic region of the USA. The field had been cropped to no-till maize. NH₃ volatilization was measured with the micro-meteorological, integrated horizontal flux (IHF) method, and denitrification with a core incubation method using acetylene inhibition. An early-winter surface application (5 December 1996; 88 m³ ha⁻¹ supplying 91 kg NH ₄ ⁺ -N ha⁻¹) was either unincorporated or immediately incorporated. NH₃ volatilization was measured from the unincorporated application, and denitrification from both slurry treatments and appropriate control soils. Total NH₃ loss from the unincorporated slurry application was 19% of applied NH ₄ ⁺ -N; temperatures were cool (4–6 °C), and 25 mm of rain fell within 24 h of application. For 3 months, enhanced denitrification occurred from the two slurry treatments, with generally higher rates from the incorporated slurry. Total net denitrification loss from the surface-applied and incorporated slurry treatments was, respectively, 11 and 17% of applied NH ₄ ⁺ -N. Denitrification loss over the winter/early-spring period was appreciable but not substantial, even where NH₃ volatilization was restricted by immediate incorporation. From the spring application (30 April 1997, 39 m³ ha⁻¹ supplying 51 kg NH ₄ ⁺ -N ha⁻¹), total NH₃ loss was 71% of applied NH ₄ ⁺ -N. These NH₃ volatilization loss data and the similarity of climate suggest that NH₃ loss factors from recent NW European work are likely to be generally applicable in the mid-Atlantic region. NH₃ volatilization from the spring application was also measured using the Z-instrument (Z_INST) approach, and with a system of small wind tunnels. A comparative assessment of the three methods is reported.     

Microbial community of aerobic granules for ammonium and sulphide removal in a sequencing batch reactor

Aerobic granules for sulphide and ammonium removal were cultivated in a sequencing batch reactor, and the microbial community of the aerobic granules was investigated by denaturing gradient gel electrophoresis. The loading rate increased from 0.15 to 0.9 kg S2? m?3 d?1, and the removal efficiencies of sulphide, chemical oxygen demand, and NH4 +-N were higher than 99, 80, and 98%, respectively. However, sludge settleability became poorer when the loading rate exceeded 0.3 kg S2? m?3 d?1. The denitrifying bacteria in the aerobic granules were Thauera sp., Pseudomonas alcaligenes, and uncultured planctomycetes, indicating that multiple N-removing processes occurred simultaneously in the aerobic granules. These processes could include nitrification and denitrification, aerobic denitrification, and anaerobic ammonia oxidation. Sludge settleability became poorer because of the overgrowth of uncultured Thiothrix sp.     

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.     

Screening and characterization of an aerobic nitrifying-denitrifying bacterium from activated sludge

Taking advantage of the good biocompatibility and high efficiency of nitrogen removal with microbes, nitrifying and denitrifying bacteria, are becoming increasingly more widely used for wastewater treatment and prevention of eutrophication. In this research, an aerobic nitrifying-denitrifying bacterium was successfully screened from activated sludge and identified as Pseudomonas sp. (CCTCC No M2010209) by the 16S rDNA sequence. The activity verification confirmed its nitrifying-denitrifying capability of removing ammonium, nitrate and nitrite nitrogen. The types of carbon sources and carbon-nitrogen ratio greatly influenced the removal efficiency of NH₄ ⁺-N and NO₃ ⁻-N. When the initial concentrations of NH₄ ⁺-N and NO₃ ⁻-N in synthetic wastewater were less than 70 and 50 mg/L, the nitrogen removal rates reached 94 and 90% in 9 h, respectively. Preliminary comparisons of nitrogen removal capacity between this isolate and other commercial preparations in the treatment of synthetic wastewater revealed its promising potential to be used in the actual wastewater treatment.     

Dynamic variations of microbial community structure in Myriophyllum aquaticum constructed wetlands in response to different NH4+-N concentrations

Constructed wetlands (CWs) have received increasing attentions for their N removal performances, especially regarding NH₄⁺-N. Different influent NH₄⁺-N concentration may influence N removal efficiency in practice, while the effects of different NH₄⁺-N concentrations on microorganisms removing N in CWs are poorly understood. In this study, surface flow CWs planted with Myriophyllum (M). aquaticum were established to investigate the influences of different NH₄⁺-N concentrations on the composition, structure, and interactions of microbial community. Our findings suggested 105 mg/L NH₄⁺-N CWs achieved highest N removal rate, removing 89.30 % NH₄⁺-N and 92.34 % TN from the influent. The results of real-time quantitative polymerase chain reactions (qPCR) indicated abundances of nitrifying genes (nxrA) and denitrifying genes (narG, nirS, nirK, and nosZ) were increased by increasing NH₄⁺-N concentrations, and the strongest effects were observed in narG (8-fold) and nosZ genes (11-fold). Different NH₄⁺-N concentrations was proved to alter composition and structure of microbial communities via high-throughput sequencing, e.g. denitrifiers including Brevendomonas.sp, Dokdonella.sp and Rhodococcus.sp were enriched obviously with increasing NH₄⁺-N concentrations. In addition, network showed interactions among microbial populations and positive interactions were dramatically shifted and enhanced by increasing NH₄⁺-N concentrations.     

气提式内循环硝化反应器运行性能的研究

气提式内循环反应器具有很好的生物硝化性能,能承受高进水氨浓度（78.49mmol/L）,具有高容积转化效率（163.18 mmol/L·d）,运行性能稳定（氨去除率保持在94.42%以上）。在气提式内循环反应器的运行过程中,可产生硝化颗粒污泥。颗粒污泥开始出现的时间约为45d,颗粒污泥的粒径平均值0.83 mm,沉降速度55.53m/h,氨氧化活性0.95mmol (NH⁺₄-N)/g(VS)·d。硝化颗粒污泥也具有厌氧氨氧化活性,氨氧化速率0.23mmol (NH⁺₄-N)/g(VS)·d,亚硝酸还原速率0.24mmol (NO^-₂-N)/g(VS)·d。     

Biological nitrification–denitrification with alternating oxic and anoxic operations using aerobic granules

Efficient nitrification and denitrification of wastewater containing 1,700 mgl⁻¹ of ammonium-nitrogen was achieved using aerobic granular sludge cultivated at medium-to-high organic loading rates. The cultivated granules were tested in a sequencing batch reactor (SBR) fed with 6.4 or 10.2 kg NH₄⁺-N m⁻³ day⁻¹, a loading significantly higher than that reported in literature. With alternating 2 h oxic and 2 h anoxic operation (OA) modes, removal rate was 45.5 mg NH₄⁺-N g⁻¹ volatile suspended solids⁻¹ h⁻¹ at 6.4 kg NH₄⁺-N m⁻³ day⁻¹ loading and 41.3 ± 2.0 at 10.2 kg NH₄⁺-N m⁻³ day⁻¹ loading. Following the 60 days SBR test, granules were intact. The fluorescence in situ hybridization and confocal laser scanning microscopy results indicate that the SBR-OA granules have a distribution with nitrifers outside and heterotrophs outside that can effectively expose functional strains to surrounding substrates at high concentrations with minimal mass transfer limit. This microbial alignment combined with the smooth granule surface achieved nitrification–denitrification of wastewaters containing high-strength ammonium using aerobic granules. Conversely, the SBR continuous aeration mode yielded a distribution with nitrifers outside and heterotrophs inside with an unsatisfactory denitrification rate and floating granules as gas likely accumulated deep in the granules.     

Heterotrophs grown on the soluble microbial products (SMP) released by autotrophs are responsible for the nitrogen loss in nitrifying granular sludge

In this work, nitrogen loss in the nitrite oxidation step of the nitrification process in an aerobic‐granule‐based reactor was characterized with both experimental and modeling approaches. Experimental results showed that soluble microbial products (SMP) were released from the nitrite‐oxidizing granules and were utilized as a carbon source by the heterotrophs for denitrification. This was verified by the fluorescence in situ hybridization (FISH) analysis. Microelectrode tests showed that oxygen diffusion limitation did result in an anoxic micro‐zone in the granules and allowed sequential utilization of nitrate as an electron acceptor for heterotrophic denitrification with SMP as a carbon source. To further elucidate the nitrogen loss mechanisms, a mathematic model was formulated to describe the growth of nitrite oxidizers, the formation and consumption of SMP, the anoxic heterotrophic growth on SMP and nitrate, as well as the oxygen transfer and the substrate diffusion in the granules. The results clearly indicate that the heterotrophs grown on the SMP released by the autotrophs are responsible for the nitrogen loss in the nitrifying granules, and give us a better understanding of the aerobic granules for nitrogen removal. Biotechnol. Bioeng. 2011;108: 2844–2852. © 2011 Wiley Periodicals, Inc.     

Formation and long-term stability of nitrifying granules in a sequencing batch reactor

The formation and long-term stability of nitrifying granules in a sequencing batch reactor was investigated in this study. The results showed that nitrifying granules with a size of 240 microm and SVI of 40 ml g(-1) were formed on day 21 at a settling time of 10 min. Maintaining settling time at 15 min from day 57 to 183 did not affect the physical characteristics of sludge and the fraction of suspended floc in the sludge. In addition, nitrifying granules could tolerate the fluctuations of nitrogen loading rate from 0.72 to 1.8 g l(-1)d(-1) during 2 months without the change of physical characteristics. However, it was observed that complete nitrification to nitrate and partial nitrification to nitrite by sludge converted each other corresponding to the change of the influent NH4+-N concentration. Thus, an appropriate method is needed to maintain a stable complete nitrification or partial nitrification under the conditions with changing influent NH4+-N concentrations and nitrogen loading rates.     

Enrichment and granulation of Anammox biomass started up with methanogenic granular sludge

A sequencing batch reactor (SBR) seeded with methanogenic granular sludge was started up to enrich Anammox (Anaerobic Ammonium Oxidation) bacteria and to investigate the feasibility of granulation of Anammox biomass. Research results showed that hydraulic retention time (HRT) was an important factor to enrich Anammox bacteria. When the HRT was controlled at 30 days during the initial cultivation, the SBR reactor presented Anammox activity at t = 58 days. Simultaneously, the methanogenic granular sludge changed gradually from dust black to brown colour and its diameter became smaller. At t = 90 days, the Anammox activity was further improved. NH₄⁺-N and NO₂⁻N were removed simultaneously with higher speed and the maximum removal rates reached 14.6 g NH₄⁺-N /(m³ _reactor·day) and 6.67 g NO₂⁻-N /(m³ _reactor·day), respectively. Between t = 110 days and t = 161 days, the nitrogen load was increased to a HRT of 5 days (70 mg/l NH₄⁺ and 70 mg/l NO₂), the removal rates of ammonium and nitrite were 60.6% and 62.5% respectively. The sludge changed to red and formed Anammox granulation with high nitrogen removal activity.     

Simultaneous ammonium-nitrogen and copper removal, and copper recovery using nitrifying biofilm from the ultra-compact biofilm reactor

Simultaneous ammonium-nitrogen (NH(4)(+)-N) and copper removal, and copper recovery in synthetic wastewater using nitrifying biofilm from an ultra-compact biofilm reactor (UCBR) was demonstrated in batch studies, which consisted of three phases: Phase 1 for NH(4)(+)-N and copper removals, Phase 2 for copper recovery, and Phase 3 for NH(4)(+)-N removal. The results showed that more than 96.3% of copper was removed within 60min, while 60.1% of the adsorbed copper was recovered through rinsing the biofilms with 0.1mM of ethylenediaminetetraacetic acid (EDTA). The nitrifying biofilm was able to adsorb 0.245mg of copper/g of biofilms. After recovery treatment, 29.4% of copper remained bound within the nitrifying biofilms. No significant inhibitory effects towards NH(4)(+)-N removal in the presence of 0.92mg copper/L was noted in Phase 1 compared with the control test. However, lower initial pH condition in the recovery process and the accumulation of copper on the biofilm led to 50% inhibition on NH(4)(+)-N removal efficiency in the subsequent phase.     

Enhanced denitrifying phosphorous removal in a novel anaerobic/aerobic/anoxic (AOA) process with the diversion of internal carbon source

A novel anaerobic/aerobic/anoxic (AOA) process is proposed to realize denitrifying phosphorous removal in this study, and the characteristic of the AOA process is transferring part of the anaerobic mixed liquor to the post-anoxic zone for providing the carbon source needed for denitrification. The AOA process was operated for 3 months, and the average removal efficiencies of NH4+-N, TN and PO4(3-)-P were 93.0±3.1%, 70.3±2.9% and 87.3±11.8%, respectively. A mass balance analysis indicated that 0.49±0.02 g VSS(-1) d(-1) of PO4(3-)-P and 0.23±0.04 g VSS(-1) d(-1) of NO3--N were simultaneously removed in the anoxic zone, and it is speculated that denitrifying phosphorous removal occurred in the AOA process. Furthermore, 0.24±0.06 g VSS(-1) d(-1) of TN was removed in the aerobic zone via simultaneous nitrification and denitrification (SND). The results demonstrate that the multi-zone structure of the AOA process favors the enhancement of denitrifying phosphorous removal and SND for municipal wastewater treatment.     

Nitrification and denitrification in a system of reciprocating jet bioreactor

Concept of separation of stages coupled with novel design of reciprocating jet bioreactor have been incorporated in this research for the development of high efficiency treatment system for contaminated wastewaters.Evaluation of pilot plant data reveals that a three stage reciprocating jet bioreactor system could be effectively employed for nitrification and denitrification of highly polluted wastewater obtained from Berlin wastewater treatment plant. Such a system with COD destruction stage (residence time 1–3 hours) followed by nitrification stage (residence time 3–4.5 hours) and denitrification stage (residence time 0.3 hours) gives COD destruction rate upto 58 kg COD/(m³ day), nitrification rate upto 3.2 NH ₄ ⁺ -N/(m³ day) and denitrification rate upto 28 kg NO ₃ ^– -N/(m³ day) while providing COD, NH ₄ ⁺ -N and NO ₃ ^– -N conversion of more than 90%.Nitrification and denitrification of wastewater at such a short residence time is possible mainly due to the employment of reciprocating jet bioreactor system.Paper presented at the Third Joint Schlesinger Seminar on Transport phenomena and processes in biological systems, Technion — Israel Institute of Technology, Haifa, Israel, May 8–9, 1990     

Organics and nitrogen removal and sludge stability in aerobic granular sludge membrane bioreactor

Novel aerobic granular sludge membrane bioreactor (GMBR) was established by combining aerobic granular sludge technology with membrane bioreactor (MBR). GMBR showed good organics removal and simultaneous nitrification and denitrification (SND) performances for synthesized wastewater. When influent total organic carbon (TOC) was 56.8-132.6 mg/L, the TOC removal of GMBR was 84.7-91.9%. When influent ammonia nitrogen was 28.1-38.4 mg/L, the ammonia nitrogen removal was 85.4-99.7%, and the total nitrogen removal was 41.7-78.4%. Moreover, batch experiments of sludge with different particle size demonstrated that: (1) flocculent sludge under aerobic condition almost have no denitrification capacity, (2) SND capacity was caused by the granular sludge, and (3) the denitrification rate and total nitrogen removal efficiency were enhanced with the increased particle size. In addition, study on the sludge morphology stability in GMBR showed that, although some granular sludge larger than 0.9 mm disaggregated at the beginning of operation, the granular sludge was able to maintain the stability of its granular morphology, and at the end of operation, the amount of granular sludge (larger than 0.18 mm) stabilized in GMBR was more than 56-62% of the total sludge concentration. The partial disaggregation of large granules is closely associated with the change of operating mode from sequencing batch reactor (SBR) system to MBR system.