Kinetics of nitrification and denitrification reactions

Nitrification and denitrification are important microbiological reactions of nitrogen. In this work, the kinetics of these reactions have been investigated based on a Monod-type expression involving two growth limiting substrates: ammonium nitrogen and dissolved oxygen for nitrification and nitrate nitrogen and dissolved organic carbon for denitrification. The kinetic constants and yield coefficients were evaluated for both these reactions. Past experimental work was used to determine the constants for the nitrification reaction. For the denitrification reaction, experiments were performed in a stirred tank reactor under conditions such that only one substrate was growth limiting. Steady-state values of the substrate concentrations in the reactor were determined at various dilution rates. These data were analyzed to obtain the kinetic and stoichiometric constants. From these constants it was concluded that in the range of nitrate nitrogen concentrations encountered in waste water, the denitrification reaction can be considered a first-order reaction. It was also found that three times as much organic carbon is required as nitrate nitrogen for complete nitrogen removal.     

限氧自养硝化-反硝化生物脱氮新技术

限氧自养硝化—反硝化是部分硝化与厌氧氨氧化相耦联的生物脱氮反应过程,通过严格控制溶解氧在0．1～0．3mg·L^-1,实现硝化反应控制在亚硝酸阶段,然后以硝化阶段剩余的NH4^+作为电子供体,在厌氧条件下实现反硝化,该反应过程是完全的自养硝化—反硝化过程,具有能耗低、脱氮效率高、反应系统占地面积小等优点,适用于处理COD／NH4^+—N低的废水,是一种非常有应用前景的生物脱氮技术,文中详细介绍了限氧自养硝化—反硝化生物脱氮反应过程的研究进展,讨论了其微生物学机理及应用前景。     

New pathways for ammonia conversion in soil and aquatic systems

Ammonia conversion processes are essential for most soil and aquatic systems. Under natural conditions, the many possible reactions are difficult to analyze. For example, nitrification and denitrification have long been regarded as separate phenomena performed by different groups of bacteria in segregated areas of soils, sediments or aquatic systems sequentially in time. It has now been established that strict segregation in place and time of the two processes is not necessary and that both denitrifiers and nitrifiers have versatile metabolisms. However, the rates described for aerobic denitrifiers are very low compared to the rates observed under anoxic conditions. Also the rates of nitrifier denitrification are quite low, indicating that these conversions may not play an important role under natural conditions. In addition, these processes often result in the emission of quite large amounts of undesirable products, NO and N₂O. Heterotrophic nitrification might be of relevance for systems, that contain a high carbon to nitrogen ratio. Recently, a novel process (Anammox) has been discovered in which ammonium serves as the electron donor for denitrification of nitrite into dinitrogen gas. ¹⁵N labeling studies showed that hydrazine and hydroxylamine were important intermediates in this process. Enrichment cultures on ammonium, nitrite and bicarbonate resulted in the dominance of one morphotypical microorganism. The growth rate of the cultures is extremely low (doubling time 11 days), but the affinity for ammonium and nitrite and the conversion rates (9.2 10^–4 mol kg^–1 s^–1) are quite high. Some of the reported high nitrogen losses in soil and aquatic systems might be attributed to anaerobic ammonium oxidation. In addition, this conversion offers new opportunities for nitrogen removal, when it is combined with recently developed processes for partial nitrification.     

湖泊氮素氧化及脱氮过程研究进展

自然界中氮的生物地球化学循环主要由微生物驱动,由固氮作用、硝化作用、反硝化作用和氨化作用来完成。过去数十年间,随着异养硝化、厌氧氨氧化和古菌氨氧化作用的发现,人们对环境中氮素循环认识逐步深入,提出了多种脱氮途径新假说。对湖泊生态系统中氮素的输入、输出及其在水体、沉积物和水土界面的迁移转化过程进行了概括,对湖泊生态系统中反硝化和厌氧氨氧化脱氮机理及脱氮效率的最新研究进展进行了探讨,并对以后的氮素循环研究进行了展望。     

Nitrogen dynamics in the Westerschelde estuary (SW Netherlands) estimated by means of the ecosystem model MOSES

A tentative nitrogen budget for the Westerschelde (SW Netherlands) is constructed by means of a simulation model with thirteen spatial compartments. Biochemical and chemical processes in the water column are dynamically modeled; fluxes of dissolved constituents across the water-bottom interface are expressed by means of diagenetic equations.The model is calibrated on a large amount of observed variables in the estuary (1980–1986) with relatively fine temporal and spatial detail. Additional constraints are imposed by the stoichiometric coupling of carbon, nitrogen and oxygen flows and the required conservation of mass. The model is able to reproduce rather well the observed distributions of nitrate, ammonium, oxygen and Kjeldahl nitrogen both in time and space. Also, model output of biochemical oxygen demand and total organic carbon falls within observed ranges.By far the most pervasive process in the nitrogen cycle of the estuary is nitrification which mainly takes place in the water column of the upper estuarine part. On average about three times as much nitrate is leaving the estuary at the sea side compared to what enters from the river and from waste discharges. Ammonium on the other hand is consumed much faster (nitrification) than it is regenerated and only about one third of the total import leaves the estuary at the sea side. The budget for detrital nitrogen reveals import from the river, from wastes and from the sea. Phytoplankton uptake of inorganic nitrogen is negligible in the model.About 21% of total nitrogen, 33% of inorganic nitrogen, is removed from the estuary (mainly to the atmosphere through denitrification) and the load of nitrogen net exported to the sea amounts to about 51 000 tonnes per year. Total denitrification in our model is lower than what was estimated in the literature from the late seventies, where a nitrogen removal up to 40–50% of the total inorganic load was reported. Part of the differences could be methodological, but inspection of the nutrient profiles that led to these conclusions show them to be different to the ones used in our study. The oxygen deficient zone has moved upstream since the late seventies, entrailing the zone of denitrification into the riverine part of the Schelde. The nitrification process now starts immediately upon entering the estuary.     

以N-甲基吡咯烷酮(NMP)为电子供体进行反硝化的可行性及实验研究

【目的】利用N-甲基吡咯烷酮(N-methylpyrrolidone, NMP)作为电子供体进行反硝化实验,以实现废水资源化。【方法】分别将NMP废水和葡萄糖作为电子供体加入到模拟的城市污水处理尾水中进行反硝化,比较2种电子供体去除硝酸盐的规律。同时考查NMP在反硝化过程中的氮素释放规律,并对所释放的氮素进行后续处理。最后再对它们作为电子供体时的反硝化污泥采用高通量测序,从微生物群落的角度分析NMP作为电子供体时其作用机理是否相同。【结果】当以NMP为电子供体时,硝酸盐氮的去除速率比葡萄糖为电子供体时要快67%。在8 h的反硝化结束后,剩余的硝酸盐氮、累积的亚硝酸盐氮和NMP本身所释放氨氮之和的总氮,与葡萄糖为电子供体时相近。【结论】NMP废水可以作为电子供体用于城镇污水处理厂的深度脱氮。对2种碳源所驯化的反硝化污泥样品进行高通量分析表明,NMP与葡萄糖作为电子供体用于反硝化反应时,相关的作用机理是不同的。该项研究结果对利用含氮杂环化合物作为电子供体进行反硝化具有重要的理论指导意义。     

Modeling nitrogen removal in water hyacinth ponds receiving effluent from waste stabilization ponds

We developed a dynamic model to predict nitrogen removal in water hyacinth ponds (WHPs) receiving effluent from waste stabilization ponds (WSPs). The model is based on the biofilm reaction on the root surface of plant and pond walls. The model consists of mass balances of six main substrates including: particulate organic nitrogen (PON), dissolved organic nitrogen (DON), ammonium (NH₄⁺), nitrite and nitrate (NOx), soluble chemical oxygen demand (SCOD), and particulate chemical oxygen demand (PCOD). The model, incorporating major nitrogen transformation mechanisms such as hydrolysis, mineralization, and nitrification–denitrification, accounts also for carbon consumption and plant uptake. The model's application to a pilot plant showed good agreement between measured and predicted values. According to the modeling results, in the WHPs, nitrification and denitrification were the predominant nitrogen removal processes occurring simultaneously. Temperature and hydraulic retention time (HRT) had a profound effect on the performance of nitrogen removal while an algae biomass (PCOD) accumulated in the WHPs, was a useful carbon source for denitrification.     

Effect of sulfide concentration on autotrophic denitrification from nitrate and nitrite in vertical fixed-bed reactors

Autotrophic denitrification coupled with sulfide oxidation represents an interesting alternative for the simultaneous removal of nitrate/nitrite and sulfide from wastewaters. The applicability of such bioprocess is especially advantageous for the post treatment of effluents from anaerobic reactors, since they usually produce sulfides, which can be used as endogenous electron donor for autotrophic denitrification. This study evaluated the effect of sulfide concentration on this bioprocess using nitrate and nitrite as electron acceptors in vertical fixed-bed reactors. The results showed that intermediary sulfur compounds were mainly produced when excess of electron donor was applied, which was more evident when nitrate was used. Visual evidences suggested that elemental sulfur was the intermediary compound produced. There was also evidence that the elemental sulfur previously formed was being used when sulfide was applied in stoichiometric concentration relative to nitrate/nitrite. Nitrite was more readily consumed than nitrate. For both electron acceptors and sulfide concentrations tested, autotrophic denitrification was not affected by residual heterotrophic denitrification via endogenic activity, occurring as a minor additional nitrogen removal process.     

Extreme 15N-enrichments in 2.72-Gyr-old sediments: evidence for a turning point in the nitrogen cycle

Although nitrogen is a key element in organic molecules such as nucleic acids and proteins, the timing of the emergence of its modern biogeochemical cycle is poorly known. Recent studies on the antiquity of the nitrogen cycle and its interaction with free oxygen suggests the establishment of a complete aerobic N biogeochemical cycle with nitrification, denitrification, and nitrogen fixation at about 2.68 Gyr. Here, we report new bulk nitrogen isotope data for the 2.72 billion-year-old sedimentary succession of the Tumbiana Formation (Pilbara Craton, Western Australia). The nitrogen isotopic compositions vary widely from +8.6‰ up to +50.4‰ and are inversely correlated with the very low δ(13)C values of associated organic matter defining the Fortescue excursion (down to about -56‰). We propose that this (15)N-enrichment records the onset of nitrification coupled to the continuous removal of its derivatives (nitrite and nitrate) by denitrification. This finding implies an increase in the availability of electron acceptors and probably oxygen in the Tumbiana depositional environment, 300 million years before the oxygenation of the Earth's atmosphere.     

稻田生态系统生物硝化-反硝化作用与氮素损失

从土壤微生物生理学和土壤生物化学角度综述了稻田生态系统土壤生物硝化反硝化作用与氮素损失的研究进展,并探讨了土壤生物硝化反硝化作用在稻田生态系统氮素气态损失中的地位和重要性以及土壤生物硝化反硝化作用的测度方法的比较.     

川西亚高山针叶林不同恢复阶段土壤的硝化和反硝化作用

 采用气压过程分离(Barometric process separation, BaPS)技术对川西亚高山针叶林不同恢复 阶段土壤的总硝化和反硝化作用速率进行了测定,结果表明：川西亚高山针叶林不同恢复阶段土壤的总硝化和反硝化速率差异不显著(p<0.05),不同恢复阶段土壤总硝化作用的 Q10值 差异不显著(p<0.05);总硝化作用速率与土壤含水量呈显著正相关(p<0.05),与土 壤pH值、 土壤有机质、全氮及C/N相关不显著;不同恢复阶段土壤反硝化速率均维持在一个较低的水 平,反硝化速率与土壤中的C/N显著正相关(p<0.05),与土壤含水量、pH值、有机质及全氮相关不显著。与反硝化作用相比,硝化作用对亚高山针叶林土壤氮损失的影响可能更大     

Design and test of a generic cohort model of soil organic matter decomposition: the SOMKO model

• 1 SOMKO is a new simulation model of soil organic matter (SOM) dynamics aimed at predicting long‐term and short‐term SOM dynamics based on a mechanistic approach focusing on microbes as the key agents of decomposition.
• 2 SOM is partitioned into cohorts and chemical quality pools (classified by age and chemical composition), the microbial community processes are explicitly represented, and the C : N stoichiometric constraints are accounted for through a new mechanism of offer and demand.
• 3 The analysis of model equations shows that: (1) SOM C : N cannot decrease below microbial C : N; and (2) the nitrogen limitation of decomposition depends on SOM C : N, microbial biomass and soil mineral nitrogen. First tests of the model show good qualitative behaviour for simulating the dynamics of short‐term litter‐bag type decomposition, long‐term SOM increase, pulsed mineral nitrogen production, the priming effect due to easily decomposable carbon addition, and the effects of vegetation clearance and climate change on SOM. Simulations are in good agreement with long‐term experimental data.
• 4 SOMKO is an integrated component of the coupled soil–vegetation models within the ETEMA (European Terrestrial Ecosystem Modelling Activity) framework. Future extensions of this work include: (1) estimating microbial parameters from specific experiments; (2) spatial distribution of SOMKO in multistrata models; and (3) implementing nitrification/denitrification processes, phosphorus limitation and microfaunal activity.

     

Simultaneous nitrification,denitrification, and phosphorus removal in a lab-scale sequencing batch reactor

Simultaneous nitrification and denitrification (SND) via the nitrite pathway and anaerobic-anoxic-enhanced biological phosphorus removal (EBPR) are two processes that can significantly reduce the energy and COD demand for nitrogen and phosphorus removal. The combination of these two processes has the potential of achieving simultaneous nitrogen and phosphorus removal with a minimal requirement for COD. A lab-scale sequencing batch reactor (SBR) was operated in alternating anaerobic-aerobic mode with a low dissolved oxygen (DO) concentration (0.5 mg/L) during the aerobic period, and was demonstrated to accomplish nitrification, denitrification, and phosphorus removal. Under anaerobic conditions, COD was taken up and converted to polyhydroxyalkanoates (PHAs), accompanied by phosphorus release. In the subsequent aerobic stage, PHA was oxidized and phosphorus was taken up to <0.5 mg/L by the end of the cycle. Ammonia was also oxidized during the aerobic period, but without accumulation of nitrite or nitrate in the system, indicating the occurrence of simultaneous nitrification and denitrification. However, off-gas analysis showed that the final denitrification product was mainly nitrous oxide (N(2)O), not N(2). Further experimental results demonstrated that nitrogen removal was via nitrite, not nitrate. These experiments also showed that denitrifying glycogen-accumulating organisms (DGAOs), rather than denitrifying polyphosphate-accumulating organisms (DPAOs), were responsible for the denitrification activity.     

Nitrification contributes to winter oxygen depletion in seasonally frozen forested lakes

In lakes that experience seasonal ice cover, understanding of nitrogen–oxygen coupling and nitrification has been dominated by observations during open water, ice-free conditions. To address knowledge gaps about nitrogen–oxygen linkages under ice, we examined long-term winter data (30 + years, 2–3 sample events per winter) in 7 temperate lakes of forested northern Wisconsin, USA. Across lakes and depths, there were strong negative relationships between dissolved oxygen (DO) and the number of days since ice-on, reflecting consistent DO consumption rates under ice. In two bog lakes that routinely experience prolonged winter DO concentrations below 1.0 mg L^?1, nitrate accumulated near the ice surface mainly in late winter, suggesting nitrification may depend on biogenic oxygen from photosynthesis. In contrast, within five oligotrophic-mesotrophic lakes, nitrate accumulated more consistently over winter and often throughout the water column, especially at intermediate depths. Exogenous inputs of nitrate to these lakes were minimal compared to rates of nitrate accumulation. To produce the nitrate via in-lake nitrification, substantial oxygen consumption by ammonium oxidizing microbes would be required. Among lakes and depths that had significant DO depletion over winter, the stoichiometric nitrifier oxygen demand ranged from 1 to 25% of the DO depletion rate. These estimates of nitrifier-driven DO decline are likely conservative because we did not account for nitrate consumed by algal uptake or denitrification. Our results provide an example of nitrification at temperatures < 5 degrees C having a substantial influence on ecosystem-level nitrogen and oxygen availability in seasonally-frozen, northern forested lakes. Consequently, models of under-ice dissolved oxygen dynamics may be advanced through consideration of nitrification, and more broadly, coupled nitrogen and oxygen cycling.     

Partial nitrification—operational parameters and microorganisms involved

Nitrite is a common intermediate in at least three different oxidative or reductive biochemical pathways that occur in nature (nitrification, denitrification and dissimilatory or assimilatory nitrate reduction). Nitrite accumulation or partial nitrification has been reported in literature for decades. In engineered systems, partial nitrification is of interest as it offers cost savings in aeration as well as in the form of lesser need for addition of organic carbon as compared to the conventional denitrification. A broad range of operating parameters and factors has been reviewed in this paper which are essential for achieving partial nitrification. Of these, pH, dissolved oxygen (DO), temperature, free ammonia (FA) and nitrous acid concentrations, inhibitory compounds are important factors in achieving partial nitrification. Two groups of bacteria, namely ammonia-oxidizing bacteria (AOB) and nitrite-oxidizing bacteria (NOB) are involved in nitrification. Chemolitho-autotrophic AOB are responsible for the rate-limiting step of nitrification in a wide variety of environments, making them important in the global cycling of nitrogen. Characterization and identification of the bacterial populations in an engineered system which have been considered to be a “black box”, has been made possible by using non-cultivation based techniques such as fluorescent in situ hybridization technique (FISH), polymerase chain reaction (PCR), denaturing gradient gel electrophoresis (DGGE), Sequencing and other techniques involving quantitative chemical analyses of specific biomarkers including quinones. Accordingly, this paper also attempts to give examples of how various molecular techniques can be used for characterizing various microorganisms involved in biological nitrogen removal.     

A novel double-membrane system for simultaneous nitrification and denitrification in a single tank

A novel biological treatment system, which contains two types of membrane modules in a single tank, was developed for simultaneous nitrification and denitrification. Both of the modules were fed with the substrates on the tube side of the silicone tubes by diffusing them to the biofilms which form on the surface of the tubes. One module was fed with methanol for denitrification and the other one was fed with pure oxygen for nitrification. As a result, the interference of organic carbon on nitrification, and that of oxygen on denitrification, were both hindered by the diffusion barriers (biofilms), thereby allowing two different niches for nitrifiers and denitrifiers to coexist in a single tank. Besides saving space and the amount of alkalinity required for nitrification, this system also produced low residual chemical oxygen demand (COD) and high nitrogen removal rates (2.9-3.4 gN m-2 d-1 of surface area of membrane).     

Improved simultaneous nitrification and denitrification in a single reactor by using two different immobilization carriers with specific oxygen transfer characteristics

To maximize nitrogen utilization rates during nitrification and denitrification in a simultaneous reaction for direct nitrogen removal from ammonia–nitrogen in a single reactor, two different carriers were applied that immobilized nitrifiers and denitrifiers separately. With the optimized DO concentration and mixing ratio of immobilization carriers, ammonium–nitrogen was successfully removed as designed until the middle phase of treatment where nitrogen removal rate was higher than 83% of the theoretical value, although an imbalance between nitrification and denitrification occurred at a later phase of treatment where residual nitrate–nitrogen concentration was less than 2 mg/l. The new approach using two different carriers to immobilize nitrifiers and denitrifiers separately was proved useful for controlling both nitrification and denitrification rates, enabling the utilization of maximum treatment ability of both nitrifiers and denitrifiers in a single reactor for direct nitrogen removal from ammonium–nitrogen.     

Nitrogen-removal bioreactor capable of simultaneous nitrification and denitrification for application to industrial wastewater treatment

A bioreactor system with 30 packed gel envelopes was installed in a thermal power plant for the removal of nitrogen from ammonia-containing desulfurization wastewater. Each envelope consisted of double-sided plate gels containing Nitrosomonas europaea and Paracoccus denitrificans cells with an internal space in between for injecting an electron donor. The envelope can remove ammonia from wastewater in a single step. When the wastewater was continuously treated with the bioreactor system, it removed 95.0% of the total nitrogen in the inlet, and the total nitrogen concentration in the outlet was below 9.0 mg L⁻¹. The maximum nitrogen removal rate was 6.0 g day⁻¹ per square meter of the gel area. The maximum utilization efficiency of the injected ethanol for denitrification was 98.4%, and the total organic carbon concentration in the outflow was maintained at a low level. Since the bioreactor system could use the electron donor effectively, it was not necessary to use an additional aerobic tank to remove the electron donor and a settling tank to segregate the surplus sludge containing bacteria from wastewater. Our concept of using packed gel envelopes would be highly effective for constructing a simple and efficient nitrogen removal system capable of simultaneous nitrification and denitrification.     

Simultaneous nitrification and denitrification using stored substrate (PHB) as the electron donor in an SBR

The potential for PHB (poly-beta-hydroxybutyrate) to serve as the electron donor for effective simultaneous nitrification and denitrification (SND) was investigated in a 2-L sequencing batch reactor (SBR) using a mixed culture and acetate as the organic substrate. During the feast period (i.e., acetate present), heterotrophic respiration activity was high and nitrification was prevented due to the inability of nitrifying bacteria to compete with heterotrophs for oxygen. Once acetate was depleted the oxidation rate of PHB was up to 6 times slower than that of soluble acetate and nitrification could proceed due to the decreased competition for oxygen. The slow nature of PHB degradation meant that it was an effective substrate for SND, as it was oxidised at a similar rate to ammonium and was therefore available for SND throughout the entire aerobic period. The percentage of nitrogen removed via SND increased at lower DO concentrations during the famine period, with up to 78% SND achieved at a DO concentration of 0.5 mg L(-1). However, the increased percentage of SND at a low DO concentration was compromised by a 2-times slower rate of nitrogen removal. A moderate DO concentration of 1 mg L(-1) was optimal for both SND efficiency (61%) and rate (4.4 mmol N x Cmol x(-1) x h(-1)). Electron flux analysis showed that the period of highest SND activity occurred during the first hour of the aerobic famine period, when the specific oxygen uptake rate (SOUR) was highest. It is postulated that a high SOUR due to NH(4) (+) and PHB oxidation decreases oxygen penetration into the floc, creating larger zones for anoxic denitrification. The accumulation of nitrate towards the end of the SND period showed that SND was finally limited by the rate of denitrification. As PHB degradation was found to follow first-order kinetics (df(PHB)/dt = -0.19 x f(PHB)), higher PHB concentrations would be expected to drive SND faster by increasing the availability rate of reducing power and reducing penetration of oxygen into the floc, due to the corresponding increased SOUR. Process control techniques to accumulate higher internal PHB concentrations to improve PHB-driven SND are discussed.     

Simultaneous Nitrification and Denitrification in Aerobic Chemostat Cultures of Thiosphaera pantotropha

Thiosphaera pantotropha is capable of simultaneous heterotrophic nitrification and aerobic denitrification. Consequently, its nitrification potential could not be judged from nitrite accumulation, but was estimated from complete nitrogen balances. The maximum rate of nitrification obtained during these experiments was 93.9 nmol min⁻¹ mg of protein⁻¹. The nitrification rate could be reduced by the provision of nitrate, nitrite, or thiosulfate to the culture medium. Both nitrification and denitrification increased as the dissolved oxygen concentration fell, until a critical level was reached at approximately 25% of air saturation. At this point, the rate of (aerobic) denitrification was equivalent to the anaerobic rate. At this dissolved oxygen concentration, the combined nitrification and denitrification was such that cultures receiving ammonium as their sole source of nitrogen appeared to become oxygen limited and the nitrification rate fell. It appeared that, under carbon-and energy-limited conditions, a high nitrification rate was correlated with a reduced biomass yield. To facilitate experimental design, a working hypothesis for the mechanism behind nitrification and denitrification by T. pantotropha was formulated. This involved the basic assumption that this species has a “bottleneck” in its cytochrome chain to oxygen and that denitrification and nitrification are used to overcome this. The nitrification potential of other heterotrophic nitrifiers has been reconsidered. Several species considered to be “poor” nitrifiers also simultaneously nitrify and denitrify, thus giving a falsely low nitrification potential.