Metabolism of nitric oxide and nitrous oxide during nitrification and denitrification in soil at different incubation conditions

Abstract NO production and consumption rates as well as N₂O accumulation rates were measured in a loamy cambisol which was incubated under different conditions (i.e. soil moisture content, addition of nitrogen fertilizer and/or glucose, aerobic or anaerobic gas phase). Inhibition of nitrification with acetylene allowed us to distinguish between nitrification and denitrification as sources of NO and N₂O. Under aerobic conditions untreated soil showed very low release of NO and N₂O but high consumption of NO. Fertilization with NH₄⁺ or urea stimulated both NO and N₂O production by nitrification. Addition of glucose at high soil moisture contents led to increased N₂ and N₂O production by denitrification, but not to increased NO production rates. Anaerobic conditions, however, stimulated both NO and N₂O production by denitrification. The production of NO and N₂O was further stimulated at low moisture contents and after addition of glucose or NO₃⁻. Anaerobic consumption of NO by denitrification followed Michaelis-Menten kinetics and was stimulated by addition of glucose and NO₃⁻. Aerobic consumption of NO followed first-order kinetics up to mixing ratios of at least 14 ppmv NO, was inhibited by autoclaving but not by acetylene, and decreased with increasing soil moisture content. The high NO-consumption activity and the effects of soil moisture on the apparent rates of anaerobic and aerobic production and consumption of NO suggest that diffusional constraints have an important influence on the release of NO, and may be a reason for the different behaviour of NO release vs N₂O release.     

Metabolism of nitric oxide and nitrous oxide during nitrification and denitrification in soil at different incubation conditions

Abstract NO production and consumption rates as well as N₂O accumulation rates were measured in a loamy cambisol which was incubated under different conditions (i.e. soil moisture content, addition of nitrogen fertilizer and/or glucose, aerobic or anaerobic gas phase). Inhibition of nitrification with acetylene allowed us to distinguish between nitrification and denitrification as sources of NO and N₂O. Under aerobic conditions untreated soil showed very low release of NO and N₂O but high consumption of NO. Fertilization with NH₄⁺ or urea stimulated both NO and N₂O production by nitrification. Addition of glucose at high soil moisture contents led to increased N₂ and N₂O production by denitrification, but not to increased NO production rates. Anaerobic conditions, however, stimulated both NO and N₂O production by denitrification. The production of NO and N₂O was further stimulated at low moisture contents and after addition of glucose or NO₃⁻. Anaerobic consumption of NO by denitrification followed Michaelis-Menten kinetics and was stimulated by addition of glucose and NO₃⁻. Aerobic consumption of NO followed first-order kinetics up to mixing ratios of at least 14 ppmv NO, was inhibited by autoclaving but not by acetylene, and decreased with increasing soil moisture content. The high NO-consumption activity and the effects of soil moisture on the apparent rates of anaerobic and aerobic production and consumption of NO suggest that diffusional constraints have an important influence on the release of NO, and may be a reason for the different behaviour of NO release vs N₂O release.     

畜禽粪便好氧堆肥过程氧化亚氮排放机制

好氧堆肥是实现畜禽粪便处理及资源化的有效途径,但畜禽粪便好氧堆肥过程是全球温室气体N2O的潜在来源,与全球温室效应和大气臭氧空洞等问题密切相关.随着畜禽养殖规模的扩大和畜禽粪便堆肥产量的急剧提升,畜禽粪便好氧堆肥过程N2O排放问题日趋严重,堆肥过程N2O排放机制机理研究引发学者们的关注.本文综述了畜禽粪便好氧堆肥过程中N2O的产生途径、排放规律、排放影响因素及其相关微生物学机理研究动态,总结了该过程中减排N2O的措施,并对该领域的研究趋势进行了展望.     

冻融循环期间土壤氧化亚氮排放影响因素

全球气候变化可能会提高冻融循环时间、强度以及频率, 从而可能显著影响土壤氧化亚氮(N₂O)排放。N₂O是一种重要的温室气体, 但目前对冻融循环期间土壤N₂O排放规律以及影响因素的了解还有限。为此, 该研究采用整合分析方法, 从已发表文献中收集了30篇关于冻融循环对土壤N₂O通量和累积排放量影响的文献, 探究冻融循环在不同生态系统对N₂O排放的影响, 从试验设置、土壤基本理化性质以及冻融循环格局等角度全面综合地探究其排放影响因素。该研究得出, 冻融循环能显著增加N₂O通量、N₂O累积排放量和硝化作用速率, 全球平均增幅分别为72.34%、143.25%和124.63%; 冻融循环也可增加反硝化作用速率, 全球平均增幅为162.56%; 与之相反, 冻融循环显著减少微生物生物量氮含量, 全球平均减幅为6.39%。不同生态系统土壤水热条件和基本理化性质差异可显著影响冻融循环对N₂O排放的影响。当年平均气温超过5 ℃时, 冻融循环作用可显著提高N₂O通量104.13%, 显著高于年平均气温为0-5 ℃ (25.56%)和小于0 ℃ (55.29%)时; 土壤湿度大于70%时, N₂O通量增加109.17%, 显著高于土壤湿度为50%-70% (65.67%)和小于50% (20.37%)时的通量。土壤黏粒和养分含量越高的土壤区域, 冻融循环对N₂O排放的提高幅度越大。在有植物存在时, 冻融循环可显著提高土壤N₂O通量达91.21%, 高于无植物存在时的54.43%。土壤过筛和在冻融循环期间采集土壤都会增加冻融循环对N₂O排放的影响。另外, 融化时间长, 冻结强度大和冻融循环频率高均可显著提高土壤N₂O累积排放量对冻融循环的响应。当冻结温度低于-10 ℃时, 冻融循环对土壤N₂O排放通量的增幅可达100.73%, 显著高于在冻结温度为-10- -5 ℃ (47.74%)和高于-5 ℃ (70.25%)时。主要原因是冻结强度高可促进土壤微生物和土壤结构释放更多的养分, 从而提高N₂O的产生和排放。该研究结果有助于更好地理解土壤N₂O对冻融循环的响应及其影响因素, 为更准确地预测未来全球气候变化对N₂O排放影响提供科学数据支撑。     

Sulfur-driven autotrophic denitrification of nitric oxide for efficient nitrous oxide recovery

Nitrous oxide (N₂O) was previously deemed as a potent greenhouse gas but is actually an untapped energy source, which can accumulate during the microbial denitrification of nitric oxide (NO). Compared with the organic electron donor required in heterotrophic denitrification, elemental sulfur (S⁰) is a promising electron donor alternative due to its cheap cost and low biomass yield in sulfur-driven autotrophic denitrification. However, no effort has been made to test N₂O recovery from sulfur-driven denitrification of NO so far. Therefore, in this study, batch and continuous experiments were carried out to investigate the NO removal performance and N₂O recovery potential via sulfur-driven NO-based denitrification under various Fe(II)EDTA-NO concentrations. Efficient energy recovery was achieved, as up to 35.5%–40.9% of NO was converted to N₂O under various NO concentrations. N₂O recovery from Fe(II)EDTA-NO could be enhanced by the low bioavailability of sulfur and the acid environment caused by sulfur oxidation. The NO reductase (NOR) and N₂O reductase (N₂OR) were inhibited distinctively at relatively low NO levels, leading to efficient N₂O accumulation, but were suppressed irreversibly at NO level beyond 15 mM in continuous experiments. Such results indicated that the regulation of NO at a relatively low level would benefit the system stability and NO removal capacity during long-term system operation. The continuous operation of the sulfur-driven Fe(II)EDTA-NO-based denitrification reduced the overall microbial diversity but enriched several key microbial community. Thauera, Thermomonas, and Arenimonas that are able to carry out sulfur-driven autotrophic denitrification became the dominant organisms with their relative abundance increased from 25.8% to 68.3%, collectively.     

Nitrite-driven nitrous oxide production under aerobic soil conditions: kinetics and biochemical controls

Nitrite (NO₂⁻) can accumulate during nitrification in soil following fertilizer application. While the role of NO₂⁻ as a substrate regulating nitrous oxide (N₂O) production is recognized, kinetic data are not available that allow for estimating N₂O production or soil‐to‐atmosphere fluxes as a function of NO₂⁻ levels under aerobic conditions. The current study investigated these kinetics as influenced by soil physical and biochemical factors in soils from cultivated and uncultivated fields in Minnesota, USA. A linear response of N₂O production rate () to NO₂⁻ was observed at concentrations below 60 μg N g⁻¹ soil in both nonsterile and sterilized soils. Rate coefficients (K_p) relating to NO₂⁻ varied over two orders of magnitude and were correlated with pH, total nitrogen, and soluble and total carbon (C). Total C explained 84% of the variance in K_p across all samples. Abiotic processes accounted for 31–75% of total N₂O production. Biological reduction of NO₂⁻ was enhanced as oxygen (O₂) levels were decreased from above ambient to 5%, consistent with nitrifier denitrification. In contrast, nitrate (NO₃⁻)‐reduction, and the reduction of N₂O itself, were only stimulated at O₂ levels below 5%. Greater temperature sensitivity was observed for biological compared with chemical N₂O production. Steady‐state model simulations predict that NO₂⁻ levels often found after fertilizer applications have the potential to generate substantial N₂O fluxes even at ambient O₂. This potential derives in part from the production of N₂O under conditions not favorable for N₂O reduction, in contrast to N₂O generated from NO₃⁻ reduction. These results have implications with regard to improved management to minimize agricultural N₂O emissions and improved emissions assessments.     

Gross nitrous oxide production drives net nitrous oxide fluxes across a salt marsh landscape

Sea level rise will change inundation regimes in salt marshes, altering redox dynamics that control nitrification – a potential source of the potent greenhouse gas, nitrous oxide (N₂O) – and denitrification, a major nitrogen (N) loss pathway in coastal ecosystems and both a source and sink of N₂O. Measurements of net N₂O fluxes alone yield little insight into the different effects of redox conditions on N₂O production and consumption. We used in situ measurements of gross N₂O fluxes across a salt marsh elevation gradient to determine how soil N₂O emissions in coastal ecosystems may respond to future sea level rise. Soil redox declined as marsh elevation decreased, with lower soil nitrate and higher ferrous iron in the low marsh compared to the mid and high marshes (P < 0.001 for both). In addition, soil oxygen concentrations were lower in the low and mid‐marshes relative to the high marsh (P < 0.001). Net N₂O fluxes differed significantly among marsh zones (P = 0.009), averaging 9.8 ± 5.4 μg N m^?2 h^?1, ?2.2 ± 0.9 μg N m^?2 h^?1, and 0.67 ± 0.57 μg N m^?2 h^?1 in the low, mid, and high marshes, respectively. Both net N₂O release and uptake were observed in the low and high marshes, but the mid‐marsh was consistently a net N₂O sink. Gross N₂O production was highest in the low marsh and lowest in the mid‐marsh (P = 0.02), whereas gross N₂O consumption did not differ among marsh zones. Thus, variability in gross N₂O production rates drove the differences in net N₂O flux among marsh zones. Our results suggest that future studies should focus on elucidating controls on the processes producing, rather than consuming, N₂O in salt marshes to improve our predictions of changes in net N₂O fluxes caused by future sea level rise.     

Factors promoting emissions of nitrous oxide and nitric oxide from denitrifying sequencing batch reactors operated with methanol and ethanol as electron donors

The emissions of nitrous oxide (N₂O) and nitric oxide (NO) from biological nitrogen removal (BNR) operations via nitrification and denitrification is gaining increased prominence. While many factors relevant to the operation of denitrifying reactors can influence N₂O and NO emissions from them, the role of different organic carbon sources on these emissions has not been systematically addressed or interpreted. The overall goal of this study was to evaluate the impact of three factors, organic carbon limitation, nitrite concentrations, and dissolved oxygen concentrations on gaseous N₂O and NO emissions from two sequencing batch reactors (SBRs), operated, respectively, with methanol and ethanol as electron donors. During undisturbed ultimate‐state operation, emissions of both N₂O and NO from either reactor were minimal and in the range of <0.2% of influent nitrate‐N load. Subsequently, the two reactors were challenged with transient organic carbon limitation and nitrite pulses, both of which had little impact on N₂O or NO emissions for either electron donor. In contrast, transient exposure to oxygen led to increased production of N₂O (up to 7.1% of influent nitrate‐N load) from ethanol grown cultures, owing to their higher kinetics and potentially lower susceptibility to oxygen inhibition. A similar increase in N₂O production was not observed from methanol grown cultures. These results suggest that for dissolved oxygen, but not for carbon limitation or nitrite exposure, N₂O emission from heterotrophic denitrification reactors can vary as a function of the electron donor used. Biotechnol. Bioeng. 2010; 106: 390–398. © 2010 Wiley Periodicals, Inc.     

Effect of oxic and anoxic conditions on nitrous oxide emissions from nitrification and denitrification processes

A lab-scale sequencing batch reactor fed with real municipal wastewater was used to study nitrous oxide (N(2)O) emissions from simulated wastewater treatment processes. The experiments were performed under four different controlled conditions as follows: (1) fully aerobic, (2) anoxic-aerobic with high dissolved oxygen (DO) concentration, (3) anoxic-aerobic with low DO concentration, and 4) intermittent aeration. The results indicated that N(2)O production can occur from both incomplete nitrification and incomplete denitrification. N(2)O production from denitrification was observed in both aerobic and anoxic phases. However, N(2)O production from aerobic conditions occurred only when both low DO concentrations and high nitrite concentration existed simultaneously. The magnitude of N(2) O produced via anoxic denitrification was lower than via oxic denitrification and required the presence of nitrite. Changes in DO, ammonium, and nitrite concentrations influenced the magnitude of N(2)O production through denitrification. The results also suggested that N(2)O can be produced from incomplete denitrification and then released to the atmosphere during aeration phase due to air stripping. Therefore, biological nitrogen removal systems should be optimized to promote complete nitrification and denitrification to minimize N(2)O emissions.     

Deciphering the oxygen isotope composition of nitrous oxide produced by nitrification

The ability to use δ¹⁸O values of nitrous oxide (N₂O) to apportion environmental emissions is currently hindered by a poor understanding of the controls on δ¹⁸O–N₂O from nitrification (hydroxylamine oxidation to N₂O and nitrite reduction to N₂O). In this study fertilized agricultural soils and unfertilized temperate forest soils were aerobically incubated with different ¹⁸O/¹⁶O waters, and conceptual and mathematical models were developed to systematically explain the δ¹⁸O–N₂O formed by nitrification. Modeling exercises used a set of defined input parameters to emulate the measured soil δ¹⁸O–N₂O data (Monte Carlo approach). The Monte Carlo simulations implied that abiotic oxygen (O) exchange between nitrite (NO₂^?) and H₂O is important in all soils, but that biological, enzyme‐controlled O‐exchange does not occur during the reduction of NO₂^? to N₂O (nitrifier‐denitrification). Similarly, the results of the model simulations indicated that N₂O consumption is not characteristic of aerobic N₂O formation. The results of this study and a synthesis of the published literature data indicate that δ¹⁸O–N₂O formed in aerobic environments is constrained between +13‰ and +35‰ relative to Vienna Standard Mean Ocean Water (VSMOW). N₂O formed via hydroxylamine oxidation and nitrifier‐denitrification cannot be separated using δ¹⁸O unless ¹⁸O tracers are employed. The natural range of nitrifier δ¹⁸O–N₂O is discussed and explained in terms of our conceptual model, and the major and minor controls that define aerobically produced δ¹⁸O–N₂O are identified. Despite the highly complex nature of δ¹⁸O–N₂O produced by nitrification this δ¹⁸O range is narrow. As a result, in many situations δ¹⁸O values may be used in conjunction with δ¹⁵N–N₂O data to apportion nitrifier‐ and denitrifier‐derived N₂O. However, when biological O‐exchange during denitrification is high and N₂O consumption is low, there may be too much overlap in δ¹⁸O values to distinguish N₂O formed by these pathways.     

Diurnal patterns of denitrification, oxygen consumption and nitrous oxide production in rivers measured at the whole-reach scale

1. Denitrification, net oxygen consumption and net nitrous oxide flux to the atmosphere were measured in three small rivers (discharge approximately 2–27 m³ s^?1) at the whole reach scale during Spring and Summer, 2002. Two of these rivers (Iroquois River and Sugar Creek in north‐west Indiana – north‐east Illinois, U.S.A.) drained agricultural catchments and the other (Millstone River in central New Jersey, U.S.A.) drained a mixed suburban–agricultural catchment. 2. Denitrification, oxygen consumption and N₂O flux were measured based on net changes in dissolved gas concentrations (N₂, O₂, and N₂O) during riverine transport, correcting for atmospheric exchange. On each date, measurements were made during both light and dark periods. 3. Denitrification rates in these rivers ranged from 0.31 to 15.91 mmol N m^?2 h^?1, and rates within each river reach were consistently higher during the day than during the night. This diurnal pattern could be related to cyclic patterns of nitrification driven by diurnal variations in water column pH and temperature. 4. Oxygen consumption ranged from 2.56 to 241 mmol O₂ m^?2 h^?1. In contrast to denitrification, net oxygen consumption was generally higher during the night than during the day. 5. River water was consistently supersaturated with N₂O, ranging from 102 to 209% saturated. Net flux of N₂O to the atmosphere ranged from 0.4 to 60 μmol N m^?2 h^?1. Net flux of N₂O was generally higher at night than during the day. The high flux of N₂O from these rivers strengthens the argument that rivers are an important contributor to anthropogenic emissions of this greenhouse gas.     

Nitrification and nitrous oxide production potentials in aerobic soil samples from the soil profile of a Finnish coniferous site receiving high ammonium deposition

Abstract Using aerobic soil slurry technique nitrification and nitrous oxide production were studied in samples from a pine site in Western Finland. The site received atmospheric ammonium deposition of 7–33 kg N ha⁻¹ a⁻¹ from a mink farm. The experiments with soil slurries showed that the nitrification potential in the litter layer was higher at pH 6 than at pH 4. However, the nitrification potentials in the samples from the organic and mineral horizons at pH 6 and 4 were almost equal. Also N₂O was produced at a higher rate at pH 6 than at pH 4 in slurries of the litter layer samples. The reverse was true for samples from the organic and mineral horizons. The highest N₂O production and nitrification rates were measured in the suspensions of litter layer samples. Nitrification activity in field-moist soil samples was lower than the activity in the slurries indicating that the availability of ammonium limited nitrification in these soils. Acetylene (2.5 kPa) retarded nitrification activity (70-–100%) and N₂O production (40 – 90%) in soil slurries. Acetylene inhibited the N₂O production by 40–60% during the first 3 days after its addition to field-moist samples incubated in aerobic atmosphere. After 3 days the inhibition became much lower (4–5%). The results indicate that, in soil profiles of boreal coniferous forests receiving ammonium deposition, chemolithotrophic nitrification may have importance in the N₂O production, and that changes in soil pH affect differently nitrification as well as N₂O production in litter and deeper soil layers.     

The production and emission of nitrous oxide from headwater streams in the Midwestern United States

The emission of nitrous oxide (N₂O) from streams draining agricultural landscapes is estimated by the Intergovernmental Panel on Climate Change (IPCC) to constitute a globally significant source of this gas to the atmosphere, although there is considerable uncertainty in the magnitude of this source. We measured N₂O emission rates and potential controlling variables in 12 headwater streams draining a predominantly agricultural basin on glacial terrain in southwestern Michigan. The study sites were nearly always supersaturated with N₂O and emission rates ranged from ?8.9 to 266.8 μg N₂O‐N m^?2 h^?1 with an overall mean of 35.2 μg N₂O‐N m^?2 h^?1. Stream water NO₃^? concentrations best‐predicted N₂O emission rates. Although streams and agricultural soils in the basin had similar areal emission rates, emissions from streams were equivalent to 6% of the anthropogenic emissions from soils because of the vastly greater surface area of soils. We found that the default value of the N₂O emission factor for streams and groundwater as defined by the IPCC (EF5‐g) was similar to the value observed in this study lending support to the recent downward revision to EF5‐g. However, the EF5‐g spanned four orders of magnitude across our study sites suggesting that the IPCC's methodology of applying one emission factor to all streams may be inappropriate.     

Production of nitrous oxide in artificially flooded and drained soils

Production of nitrous oxide (N₂O) was studied in one peaty and one sandy soil undergoing wetting and drying cycles. The background concentration of N₂O in the soil was compared with the N₂O produced during 4 hours of incubation with and without addition of acetylene. The concentration of N₂O in the soil under flooded conditions was relatively stable, and net consumption of N₂O was observed as often as net production. The reference area and drained soils showed somewhat different patterns compared to the flooded soils, which was probably an effect of intermediate soil water conditions. During flooding, the nitrous oxide made up less than 1% of total denitrification on 50% and 54% of the sampling occasions for the peaty and the sandy soil, respectively, and N₂O/(N₂O+N₂)-ratios exceeded 0.2 on only 6% and 3% of the sampling occasions. Under drained conditions and in the reference areas, the ratios showed a more even frequency distribution. Grouping the nitrous oxide production data for different seasons and field conditions, we found few seasonal trends. At the sandy site, mean production of N₂O was larger during the winter months. There were weak correlations between N₂O production and floodwater nitrate concentration, and between N₂O production and soil temperature. N₂O production in the reference area varied between consumption and 4.6 kg N ha^–1 month^–1 and in flooded and drained soil between consumption and 2.6 kg N ha^–1 month^–1.     

Using kinetics and modeling to predict denitrification fluxes in elemental-sulfur-based biofilms

Elemental sulfur (S⁰) can serve as an electron donor for water and wastewater denitrification, but few researchers have addressed the kinetics of S ⁰–based reduction of nitrate (NO ₃ ⁻), nitrite (NO ₂ ⁻), and nitrous oxide (N ₂O). In addition, S ⁰-based denitrifying biofilms are counter-diffusional. This is because the electron donor (S ⁰) is supplied from the biofilm attachment surface while the acceptor, for example, NO ₃ ⁻, is supplied from the bulk liquid. No existing mathematical model for S ⁰-based denitrification considers this behavior. In this study, batch tests were used to determine the kinetic parameters for the reduction of NO ₃ ⁻, NO ₂ ⁻, and N ₂O. Additionally, a biofilm model was developed to explore the effects of counter-diffusion on overall fluxes, that is, the mass of NO ₃ ⁻ or NO ₂ ⁻ removed per unit biofilm support area per unit time. The maximum specific substrate utilization rates () for NO ₃ ⁻, NO ₂ ⁻, and N ₂O were 3.54, 1.98, and 6.28 g N g COD ⁻¹·d ⁻¹, respectively. The maximum specific growth rates () were 0.71, 1.21, and 1.67 d ⁻¹ for NO ₃ ⁻ to NO ₂ ⁻, NO ₂ ⁻ to N ₂O, and N ₂O to N ₂, respectively. Results suggest that the observed NO ₂ ⁻ accumulation during S ⁰-based denitrification results from a low for NO ₂ ⁻ relative to that for NO ₃ ⁻. The high for N ₂O, relative to that for NO ₃ ⁻ and NO ₂ ⁻, suggest that little N ₂O accumulation occurs during denitrification. A counter-diffusional biofilm model was used to predict trends for NO ₃ ⁻ fluxes, and confirmed NO ₂ ⁻ accumulation in S ⁰-based denitrification biofilms. It also explains the observed detrimental effects of biofilm thickness on denitrification fluxes. This study allows a more accurate prediction of NO ₃ ⁻, NO ₂ ⁻, and N ₂O transformations in S ⁰-based denitrification.     

Nitrous oxide production, its source and distribution in urine patches on grassland on peat soil

Urine patches are considered to be important sites for nitrous oxide (N₂O) production through nitrification and denitrification due to their high concentration of nitrogen (N). The aim of the present study was to determine the microbial source and size of production of N₂O in different zones of a urine patch on grassland on peat soil. Artificial urine was applied in elongated patches of 4.5 m. Four lateral zones were distinguished and sampled for four weeks using an intact soil core incubation method. Incubation of soil cores took place without any additions to the headspace to determine total N₂O production, with acetylene addition to determine total denitrification (N₂O+N₂), and with methyl fluoride to determine the N₂O produced through denitrification.Nitrous oxide production was largest in the centre and decreased towards the edge of the patch. Maximum N₂O production was about 50 mg N m^–2 d^–1 and maximum denitrification activity was 70 mg N m^–2 d^–1. Nitrification was the main N₂O producing process. Nitrous oxide production through denitrification was only of significance when denitrification activity was high. Total N loss through nitrification and denitrification over 31 days was 4.1 g N per patch which was 2.2% of the total applied urine-N.     

Loss of nitrogen in compacted grassland soil by simultaneous nitrification and denitrification

The soils of mid-Wales in grazed permanent pasture usually exhibit stagnogley features in the top 4–10 cm even though on sloping sites, they are freely drained. Nitrogen is often poorly recovered under these conditions. Our previous studies suggest that continuing loss of available N through concurrent nitrification and denitrification might provide an explanation for poor response to fertilizer N. The work described was designated to further test this proposition. When NH ₄ ⁺ –N was applied to the surface of intact cores, equilibrated at –5kPa matric potential, about 70% of NH ₄ ⁺ –N initially present was lost within 56 days of incubation. Study of different sections of the cores showed a rise in NO ₃ ^- level in the surface 0–2.5 cm soil layer but no significant changes below this depth. The imbalance between NO ₃ ^- accumulation and NH ₄ ⁺ disappearance during the study indicated a simultaneous nitrification and denitrification in the system. Furthermore, the denitrification potential of the soil was 3–4 times greater than nitrification potential so no major build-up of NO ₃ ^- would be expected when two processes occur simultaneously in micro-scale. When nitrification was inhibited by nitrapyrin, a substantial amount of NH ₄ ⁺ –N remained in the soil and persisted till the end of the incubation. The apparent recovery of applied N increased and of the total amount of N applied, 50% more was recovered relative to without nitrapyrin. It appears that addition of nitrapyrin inhibited nitrification, and consequently denitrification, by limiting the supply of NO ₃ ^- for denitrifying organisms. Emission of N₂O from the NH ₄ ⁺ amended soil cores further confirmed that loss of applied N was the result of both nitrification and denitrification, which occurred simultaneously in adjacent sites at shallow depths. This N loss could account for the poor response to fertilizer N often observed in pastoral agriculture in western areas of the UK.