Comparison of field and laboratory measurement of denitrification and N2O production in the saturated zone of hydromorphic soils

We evaluated a new method to measure in situ denitrification under field conditions in a number of water-saturated subsoils that had a broad range of biogeochemical properties. A test solution containing ¹⁵NO₃⁻ and/or C₂H₂ was introduced to the subsoil and the subsequent production of dissolved denitrification products was measured to quantify denitrification activity. The method showed a clear production of denitrification products over time. Results were compared to laboratory-based measurements from the same soil incubated as anaerobic slurries with added ¹⁵NO₃⁻. Rates of denitrification with the in situ and the laboratory methods ranged from 1-2800 and 1-1700 μg N kg⁻¹ d⁻¹, respectively. Generally the methods gave good agreement and we consider both to be valid. However, there were some significant deviations, which we attribute to spatial heterogeneity and laboratory effects. Because the laboratory method is so much easier to perform, we suggest it should be the preferred method for large-scale studies of denitrification from the soil types we investigated. However, the two methods showed poor agreement in determining the proportion of N₂O in the total denitrification output. This was because this proportion is subject to delicate and complex control. We conclude that neither method was suitable for quantifying N₂O emission from the denitrification measurements.     

Zinc toxicity on N2O reduction declines with time in laboratory spiked soils and is undetectable in field contaminated soils

Denitrification assays in soils spiked with zinc salt have shown inhibition of the N₂O reduction resulting in increased soil N₂O fluxes with increasing soil Zn concentration. It is unclear if the same is true for environmentally contaminated soils. Net production of N₂O and N₂ was monitored during anaerobic incubations (25 °C, He atmosphere) of soils freshly spiked with ZnCl₂ and of corresponding soils that were gradually enriched with metals (mainly Zn) in the field by previous sludge amendments or by corrosion of galvanized structures. Total denitrification activity (i.e. the sum of N₂O+N₂ production rate) was not inhibited by freshly added Zn salts up to 1600 mg Zn kg⁻¹, whereas N₂O reduction decreased by 50% (EC₅₀) at total Zn concentrations of 231 mg Zn kg⁻¹ (ZEV soil) and 368 mg Zn kg⁻¹ (TM soil). In contrast, N₂O reduction was not reduced by soil Zn in any of the field contaminated soils, even at total soil Zn or soil solution Zn concentrations exceeding more than 5 times corresponding EC₅₀'s of the freshly spiked soil. The absence of adverse effects in the field contaminated soils was unrelated to soil NO₃ or organic matter concentration. Ageing (2-8 weeks) and soil leaching after spiking reduced the toxicity of Zn on N₂O reduction, either expressed as total Zn or soil solution Zn, suggesting adaptation reactions. However, no full recovery after spiking was identified at the largest incubation period in one soil. In addition, the denitrification assay performed with sewage sludge showed elevated N₂O release in Zn contaminated sludges (>6000 mg Zn kg⁻¹ dry matter) whereas this was not observed in low Zn sludge (<1000 mg Zn kg⁻¹ dry matter) suggesting limits to adaptation reactions in the sludge particles. It is concluded that the use of soils spiked with Zn salts overestimates effects on N₂O reduction. Field data on N₂O fluxes in sludge amended soils are required to identify if metals indeed promote N₂O emissions in sludge amended soils.     

Land use effects on gross nitrogen mineralization, nitrification, and N2O emissions in ephemeral wetlands

Stable ¹⁵N isotope dilution and tracer techniques were used in cultivated (C) and uncultivated (U) ephemeral wetlands in central Saskatchewan, Canada to: (1) quantify gross mineralization and nitrification rates and (2) estimate the relative proportion of N₂O emissions from these wetlands that could be attributed to denitrification versus nitrification-related processes. In-field incubation experiments were repeated in early May, mid-June and late July. Mean gross mineralization and nitrification rates (10.3 and 3.1 mg kg⁻¹ d⁻¹, respectively) did not differ between C and U wetlands on any given date. Despite these similarities, the mean NH₄⁺ pool size in the U wetlands (17.2 mg kg⁻¹) was two to three times that of the C wetlands (6.7 mg kg⁻¹) whereas the mean NO₃⁻ pool size in U wetlands (2.2 mg kg⁻¹) was less than half that of C wetlands (5.8 mg kg⁻¹). Mean N₂O emissions from the C wetlands decreased from 112.8 to 17.0 ng N₂O m² s⁻¹ from May to July, whereas mean U-wetland N₂O emissions ranged only from 31.8 to 51.1 ng N₂O m² s⁻¹ over the same period. This trend is correlated to water-filled pore space in C wetlands, demonstrating a soil moisture influence on emissions. Denitrification is generally considered the dominant emitter of N₂O under anaerobic conditions, but in the C wetlands, only 49% of the May emissions could be directly attributed to denitrification, decreasing to 29% in July. In contrast, more than 75% of the N₂O emissions from the U wetlands arose from denitrification of the soil NO₃⁻ pool throughout the season. These land use differences in emission sources and rates should be taken into consideration when planning management strategies for greenhouse gas mitigation.     

Animal treading stimulates denitrification in soil under pasture

The effects of animal treading on denitrification in a mixed ryegrass-clover pasture were studied. A single treading event of moderate or severe intensity was applied in plots during spring by using dairy cows at varying stocking rates (4.5 cows 100 m⁻² for 1.5 or 2.5 h, respectively). Treading caused a significant short-term 21 days) increase in denitrification. Denitrification rates reached a maximum of 52 g N₂O-N ha⁻¹ day⁻¹ at 8 days after severe treading compared to 2.3 g N₂O-N ha⁻¹ day⁻¹ under nil treading. Thereafter, denitrification rates declined, and were similar to non-trodden control plots after 28 days. Soil aeration, was significantly reduced by treading as expressed by water-filled porosity. In addition, soil NH₄⁺-N and NO₃⁻-N concentrations were also increased by treading. We propose that the underlying processes involved in increasing denitrification under treading were two-fold. Firstly, treading caused a temporary (e.g. 3 days after treading) reduction in soil aeration through soil physical damage, and secondly, reduced soil N utilisation prompted by reduced plant growth led to increased soil NH₄⁺-N and NO₃⁻-N availability. This study shows that treading, without the influence of other grazing animal factors (e.g. excretion), can cause a large short-term stimulation of denitrification in grass-clover pastures.     

Nitrous oxide production of heavy metal contaminated soil

Arsenic (As), lead (Pb), copper (Cu) and zinc (Zn) can be found in large concentrations in mine spills of central and northern Mexico. Interest in these heavy metals has increased recently as they contaminate drinking water and aquifers in large parts of the world and severely affect human health, but little is known about how they affect biological functioning of soil. Soils were sampled in seven locations along a gradient of heavy metal contamination with distance from a mine in San Luis Potosí (Mexico), active since about 1800 AD. C mineralization and N₂O production were monitored in an aerobic incubation experiment. Concentrations of As in the top 0-10 cm soil layer ranged from 8 to 22,992 mg kg⁻¹, from 31 to 1845 mg kg⁻¹ for Pb, from 27 to 1620 mg kg⁻¹ for Cu and from 81 to 4218 mg kg⁻¹ for Zn. There was a significant negative correlation between production rates of CO₂ and concentrations of As, Pb, Cu and Zn, and there was a significant positive correlation with pH, water holding capacity (WHC), total N and soil organic C. There was a significant negative correlation (P<0.05) between production rate of nitrous oxide (N₂O) attributed to nitrification by the inhibition method in soil incubated at 50% WHC and total concentrations of Pb and Zn, and there was a significant positive correlation (P<0.05) with pH and total N content. There was a significant negative correlation (P<0.05) between the production rate of N₂O attributed to denitrification by the inhibition method in soil incubated at 100% WHC and total concentrations of Pb, Cu and Zn, and a significant positive correlation (P<0.01) with pH; there was a significant positive correlation (P<0.05) between the production of N₂O attributed to other processes by the inhibition method and WHC, inorganic C and clay content. A negative value for production rate of N₂O attributed to nitrifier denitrification by the inhibition method was obtained at 100% WHC. The large concentrations of heavy metals in soil inhibited microbial activity and the production rate of N₂O attributed to nitrification by the inhibition method when soil was incubated at 50% WHC and denitrification when soil was incubated at 100% WHC. The inhibitor/suppression technique used appeared to be flawed, as negative values for nitrifier denitrification were obtained and as the production rate of N₂O through denitrification increased when soil was incubated with C₂H₂.     

Community composition and activities of denitrifying bacteria from adjacent agricultural soil, riparian soil, and creek sediment in Oregon, USA

We examined denitrifying bacteria from wet soils and creek sediment in an agroecosystem in Oregon, USA that received inputs of nitrogen (N) fertilizer. Our objective was to determine the variation in denitrifying community composition and activities across three adjacent habitats: a fertilized agricultural field planted to perennial ryegrass, a naturally vegetated riparian area, and creek sediment. Using C₂H₂ inhibition, denitrifying enzyme and N₂O-reductase activities were determined in short-term incubations of anaerobic slurries. A key gene in the denitrification pathway, N₂O reductase (nosZ), served as a marker for denitrifiers. Mean denitrifying enzyme activity (DEA) was similar among habitats, ranging from 0.5 to 1.8 μg N g⁻¹ dry soil h⁻¹. However, the ratio of N₂O production, without C₂H₂, to DEA was substantially higher in riparian soil (0.64±0.02; mean±standard error, n=12) than in agricultural soil (0.19±0.02) or creek sediment (0.32±0.03). Mean N₂O-reductase activity ranged from 0.5 to 3.2 μg N g⁻¹ dry soil h⁻¹, with greater activity in agricultural soil than in riparian soil. Denitrifying community composition differed significantly among habitats based on nosZ terminal-restriction fragment length polymorphisms. The creek sediment community was unique. Communities in the agricultural and riparian soil were more closely related but distinct. A number of unique nosZ genotypes were detected in creek sediment. Sequences of nosZ obtained from riparian soil were closely related to nosZ from Bradyrhizobium japonicum. Although nosZ distribution and N₂O-reductase activity differed among habitats, relationships between activity and community composition appeared uncoupled across the agroecosystem.     

Contribution of nitrification and denitrification to N2O emissions from urine patches

Urine deposition by grazing livestock causes an immediate increase in nitrous oxide (N₂O) emissions, but the responsible mechanisms are not well understood. A nitrogen-15 (¹⁵N) labelling study was conducted in an organic grass-clover sward to examine the initial effect of urine on the rates and N₂O loss ratio of nitrification (i.e. moles of N₂O-N produced per moles of nitrate produced) and denitrification (i.e. moles of N₂O produced per moles of N₂O+N₂ produced). The effect of artificial urine (52.9 g N m⁻²) and ammonium solution (52.9 g N m⁻²) was examined in separate experiments at 45% and 35% water-filled pore space (WFPS), respectively, and in each experiment a water control was included. The N₂O loss derived from nitrification or denitrification was determined in the field immediately after application of ¹⁵N-labelled solutions. During the next 24 h, gross nitrification rates were measured in the field, whereas the denitrification rates were measured in soil cores in the laboratory. Compared with the water control, urine application increased the N₂O emission from 3.9 to 42.3 μg N₂O-N m⁻² h⁻¹, whereas application of ammonium increased the emission from 0.9 to 6.1 μg N₂O-N m⁻² h⁻¹. In the urine-affected soil, nitrification and denitrification contributed equally to the N₂O emission, and the increased N₂O loss resulted from a combination of higher rates and higher N₂O loss ratios of the processes. In the present study, an enhanced nitrification rate seemed to be the most important factor explaining the high initial N₂O emission from urine patches deposited on well-aerated soils.     

Possibilities to reduce rice straw-induced global warming potential of a sandy paddy soil by combining hydrological manipulations and urea-N fertilizations

Global warming potential (GWP) of sandy paddy soils may be reduced by trade-offs between N₂O, CH₄ and CO₂ emissions. Laboratory experiments using either rice straw (1% or 0.5%) or together with urea-N (25 or 50 mg N kg⁻¹ soil) at various levels of soil water were carried out for 30 days each, to test this assumption. Waterlogging combined with urea-N increased total N₂O emissions, with greater release upon rewaterlogging (7.4 mg N kg⁻¹ soil) than experienced by removing waterlogging only. Rice straw±urea-N either emitted small amounts of N₂O or resulted in negative values at all water levels, including saturated and aerobic. Total CH₄ fluxes declined with the decreased water levels and amount of rice straw (<193 mg C kg⁻¹ soil), and also for CO₂ with the latter (<1340 mg C kg⁻¹ soil), and rewaterlogging had little influence on both. N₂O under rewaterlogged and waterlogged±urea-N, CH₄ under waterlogged with rice straw, and CO₂ for the remainder were the major contributors to GWP. Results show that waterlogging following aerobic decomposition of rice straw (1%) with urea-N, applied either at the beginning or at the end of the aerobic conditions, could decrease GWP by 56-64% and 32-42% over the sole addition of rice straw (1% and 0.5%) under waterlogged and saturated conditions, respectively.     

N2O emissions and product ratios of nitrification and denitrification as affected by freezing and thawing

Agricultural soils contribute significantly to atmospheric nitrous oxide (N₂O). A considerable part of the annual N₂O emission may occur during the cold season, possibly supported by high product ratios in denitrification (N₂O/(N₂+N₂O)) and nitrification (N₂O-N/(NO₃⁻-N+NO₂⁻-N)) at low temperatures and/or in response to freeze-thaw perturbation. Water-soluble organic materials released from frost-sensitive catch crops and green manure may further increase winter emissions. We conducted short-term laboratory incubations under standardized moisture and oxygen (O₂) conditions, using nitrogen (N) tracers (¹⁵N) to determine process rates and sources of emitted N₂O after freeze-thaw treatment of soil or after addition of freeze-thaw extract from clover. Soil respiration and N₂O production was stimulated by freeze-thaw or addition of plant extract. The N₂O emission response was inversely related to O₂ concentration, indicating denitrification as the quantitatively prevailing process. Denitrification product ratios in the two studied soils (pH 4.5 and 7.0) remained largely unaltered by freeze-thaw or freeze-thaw-released plant material, refuting the hypothesis that high winter emissions are due to frost damage of N₂O reductase activity. Nitrification rates estimated by nitrate (NO₃⁻) pool enrichment were 1.5-1.8 μg NO₃-N g⁻¹ dw soil d⁻¹ in freeze-thaw-treated soil when incubated at O₂ concentrations above 2.3 vol% and one order of magnitude lower at 0.8 vol% O₂. Thus, the experiments captured a situation with severely O₂-limited nitrification. As expected, the O₂ stress at 0.8 vol% resulted in a high nitrification product ratio (0.3 g g⁻¹). Despite this high product ratio, only 4.4% of the measured N₂O accumulation originated from nitrification, reaffirming that denitrification was the main N₂O source at the various tested O₂ concentrations in freeze-thaw-affected soil. N₂O emission response to both freeze-thaw and plant extract addition appeared strongly linked to stimulation of carbon (C) respiration, suggesting that freeze-thaw-induced release of decomposable organic C was the major driving force for N₂O emissions in our soils, both by fuelling denitrifiers and by depleting O₂. The soluble C (applied as plant extract) necessary to induce a CO₂ and N₂O production rate comparable with that of freeze-thaw was 20-30 μg C g⁻¹ soil dw. This is in the range of estimates for over-winter soluble C loss from catch crops and green manure plots reported in the literature. Thus, freeze-thaw-released organic C from plants may play a significant role in freeze-thaw-related N₂O emissions.     

Cyanobacterial inoculation of heated soils: effect on microorganisms of C and N cycles and on chemical composition in soil surface

Physiological groups of soil microorganisms, total C and N and available nutrients were investigated in four heated (350 °C, 1 h) soils (one Ortic Podsol over sandstone and three Humic Cambisol over granite, schist or limestone) inoculated (1.5 μg chlorophyll a g⁻¹ soil or 3.0 μg chlorophyll a g⁻¹ soil) with four cyanobacterial strains of the genus Oscillatoria, Nostoc or Scytonema and a mixture of them.Cyanobacterial inoculation promoted the formation of microbiotic crusts which contained a relatively high number of NH₄⁺-producers (7.4×10⁹ g⁻¹ crust), starch-mineralizing microbes (1.7×10⁸ g⁻¹ crust), cellulose-mineralizing microbes (1.4×10⁶ g⁻¹ crust) and NO₂⁻ and NO₃⁻ producers (6.9×10⁴ and 7.3×10³ g⁻¹ crust, respectively). These crusts showed a wide range of C and N contents with an average of 293 g C kg⁻¹ crust and 50 g N kg⁻¹ crust, respectively. In general, Ca was the most abundant available nutrient (804 mg kg⁻¹ crust), followed by Mg (269 mg kg⁻¹ crust), K (173 mg kg⁻¹ crust), Na (164 mg kg⁻¹ crust) and P (129 mg kg⁻¹ crust). There were close positive correlations among all the biotic and abiotic components of the crusts.Biofertilization with cyanobacteria induced great microbial proliferation as well as high increases in organic matter and nutrients in the surface of the heated soils. In general, cellulolytics were increased by four logarithmic units, amylolytics and ammonifiers by three logarithmic units and nitrifiers by more than two logarithmic units. C and N contents rose an average of 275 g C kg⁻¹ soil and 50 g N kg⁻¹ soil while the C:N ratio decreased up to 7 units. Among the available nutrients the highest increase was for Ca (315 mg kg⁻¹ soil) followed by Mg (189 mg kg⁻¹ soil), K (111 mg kg⁻¹ soil), Na (109 mg kg⁻¹ soil) and P (89 mg kg⁻¹ soil). Fluctuations of the microbial groups as well as those of organic matter and nutrients were positively correlated.The efficacy of inoculation depended on both the type of soil and the class of inoculum. The best treatment was the mixture of the four strains and, whatever the inoculum used, the soil over lime showed the most developed crust followed by the soils over schist, granite and sandstone. In the medium term there were not significant differences between the two inocula amounts tested.These results showed that inoculation of burned soils with alien N₂-fixing cyanobacteria may be a biotechnological means of promoting microbiotic crust formation, enhancing C and N cycling microorganisms and increasing organic matter and nutrient contents in heated soils.     

Denitrification in grass swards is increased under elevated atmospheric CO2

Emissions of N₂O and N₂ were measured from Lolium perenne L. swards under ambient (36 Pa) and elevated (60 Pa) atmospheric CO₂ at the Swiss free air carbon dioxide enrichment experiment following application of 11.2 g N m⁻² as ¹⁵NH₄¹⁵NO₃ or ¹⁴NH₄¹⁵NO₃ (1 at.% excess ¹⁵N). Total denitrification (N₂O+N₂) was increased under elevated pCO₂ with emissions of 6.2 and 19.5 mg ¹⁵N m⁻² measured over 22 d from ambient and elevated pCO₂ swards, respectively, supporting the hypothesis that increased belowground C allocation under elevated pCO₂ provides the energy for denitrification. Nitrification was the predominant N₂O producing process under ambient pCO₂ whereas denitrification was predominant under elevated pCO₂. The N₂-to-N₂O ratio was often higher under elevated pCO₂ suggesting that previous estimates of gaseous N losses based only on N₂O emissions have greatly underestimated the loss of N by denitrification.     

Dinitrogen and N2O emissions in arable soils: Effect of tillage, N source and soil moisture

A laboratory investigation was performed to compare the fluxes of dinitrogen (N₂), N₂O and carbon dioxide (CO₂) from no-till (NT) and conventional till (CT) soils under the same water, mineral nitrogen and temperature status. Intact soil cores (0-10 cm) were incubated for 2 weeks at 25 °C at either 75% or 60% water-filled pore space (WFPS) with ¹⁵N-labeled fertilizers (100 mg N kg⁻¹ soil). Gas and soil samples were collected at 1-4 day intervals during the incubation period. The N₂O and CO₂ fluxes were measured by a gas chromatography (GC) system while total N₂ and N₂O losses and their ¹⁵N mole fractions in the soil mineral N pool were determined by a mass spectrometer. The daily accumulative fluxes of N₂ and N₂O were significantly affected by tillage, N source and soil moisture. We observed higher (P<0.05) fluxes of N₂+N₂O, N₂O and CO₂ from the NT soils than from the CT soils. Compared with the addition of nitrate (NO₃⁻), the addition of ammonium (NH₄⁺) enhanced the emissions of these N and C gases in the CT and NT soils, but the effect of NH₄⁺ on the N₂ and/or N₂O fluxes was evident only at 60% WFPS, indicating that nitrification and subsequent denitrification contributed largely to the gaseous N losses and N₂O emission under the lower moisture condition. Total and fertilizer-induced emissions of N₂ and/or N₂O were higher (P<0.05) at 75% WFPS than with 60% WFPS, while CO₂ fluxes were not influenced by the two moisture levels. These laboratory results indicate that there is greater potential for N₂O loss from NT soils than CT soils. Avoiding wet soil conditions (>60% WFPS) and applying a NO₃⁻ form of N fertilizer would reduce potential N₂O emissions from arable soils.     

Isotopomer signatures of soil-emitted N2O under different moisture conditions—A microcosm study with arable loess soil

Soils represent the major source of the atmospheric greenhouse gas nitrous oxide (N₂O) and there is a need to better constrain the total global flux and the relative contribution of the microbial source processes. The aim of our study was to evaluate isotopomer analysis of N₂O (intramolecular distribution of ¹⁵N) as well as conventional nitrogen and oxygen isotope ratios (i) as a tool to identify N₂O production processes in soils and (ii) to constrain the isotopic fingerprint of soil-derived N₂O. We conducted a microcosm study with arable loess soil fertilized with 20 mg N kg⁻¹ of ¹⁵NO₃⁻-labeled or non-labeled ammonium nitrate. Soils were incubated for 16 d at varying moisture (55%, 75% and 85% water-filled pore space (WFPS)) in order to establish different levels of nitrification and denitrification. Dual isotope and isotopomer ratios of emitted N₂O were determined by mass spectrometric analysis of δ¹⁸O, average δ¹⁵N (δ¹⁵N^bulk) and ¹⁵N site preference (SP=difference in δ¹⁵N between the central and peripheral N-positions of the asymmetric N₂O molecule). Total rates and N₂O emission of denitrification and nitrification were determined by ¹⁵N analysis of headspace gases and soil extracts of the ¹⁵NO₃⁻ treatment. N₂O emission and denitrification increased with moisture whereas gross nitrification was almost constant. In the 55% WFPS treatment, more than half of the N₂O flux was derived from nitrification, whereas denitrification was the dominant N₂O source in the 75% WFPS and 85% WFPS treatments. Moisture conditions were reflected by the isotopic signatures since highly significant differences were observed for average δ¹⁵N^bulk, SP and δ¹⁸O. Experiment means of the 75% WFPS and 85% WFPS treatments gave negative δ¹⁵N^bulk (−18.0‰ and −34.8‰, respectively) and positive SP (8.6‰ and 15.3‰, respectively), which we explained by the fractionation during N₂O production and partial reduction to N₂. In the 55% WFPS treatment, mean SP was relatively low (1.9‰), which suggests that nitrification produced N₂O with low or negative SP. The observed influence of process condition on isotopomer signatures suggests that the isotopomer approach might be suitable for identifying N₂O source processes. However, more research is needed to determine the impact from process rates and microbial community structure. Isotopomer signatures were within the range reported from previous soil studies which supports the assumption that SP of soil-derived N₂O is lower than SP of tropospheric N₂O.     

Nitrate removal, denitrification and nitrous oxide production in the riparian zone of an ephemeral stream

Riparian zones are important features of the landscape that can buffer waterways from non-point sources of nitrogen pollution. Studies of perennial streams have identified denitrification as one of the dominant mechanisms by which this can occur. This study aimed to assess nitrate removal within the riparian zone of an ephemeral stream and characterise the processes responsible, particularly denitrification, using both in-situ and laboratory techniques. To quantify rates of groundwater nitrate removal and denitrification in-situ, nitrate was added to two separate injection-capture well networks in a perched riparian aquifer of a low order ephemeral stream in South East Queensland, Australia. Both networks also received bromide as a conservative tracer and one received acetylene to inhibit the last step of denitrification. An average of 77 ± 2% and 98 ± 1% of the added nitrate was removed within a distance of 40 cm from the injection wells (networks with acetylene and without, respectively). Based on rates of N₂O production in the network with added acetylene, denitrification was not a major mechanism of nitrate loss, accounting for only 3% of removal. Reduction of nitrate to ammonium was also not a major pathway in either network, contributing <4%. Relatively high concentrations of oxygen in the aquifer following recent filling by stream water may have reduced the importance of these two anaerobic pathways. Alternatively, denitrification may have been underestimated using the in-situ acetylene block technique. In the laboratory, soils taken from two depths at each well network were incubated with four nitrate-N treatments (ranging from ambient concentration to an addition of 15 mg N l⁻¹), with and without added acetylene. Potential rates of denitrification, N₂O production and N₂O:N₂ ratios increased with nitrate additions, particularly in shallow soils. Potential rates of denitrification observed in the laboratory were equivalent in magnitude to nitrate removal measured in the field (mean 0.26 ± 0.12 mg N kg of dry soil⁻¹ d⁻¹), but were two orders of magnitude greater than denitrification measured in the field with added acetylene. The relative importance of assimilatory vs. dissimilatory processes of nitrate removal depends on environmental conditions in the aquifer, particularly hydrology and its effects on dissolved oxygen concentrations. Depending on seasonal conditions, aquifers of ephemeral streams like the study site are likely to fluctuate between oxic and anoxic conditions; nevertheless they may still function as effective buffers. While denitrification to N₂ is a desirable outcome from a management perspective, assimilation into biomass can provide a rapid sink for nitrate, thus helping to reduce short-term delivery of nitrate downstream. Longer-term studies are needed to determine the overall effectiveness of riparian buffers associated with ephemeral streams in mitigating nitrate loads reaching downstream ecosystems.     

Differences in isotopic fractionation of nitrogen in water-saturated and unsaturated soils

Temporal variations in δ¹⁵N of NH₄⁺ and NO₃⁻ in water-saturated and unsaturated soils were examined in a laboratory incubation study. Ammonium sulfate (δ¹⁵N=−2.6‰) was added to 25 g samples of soil at concentrations of 160 mg N kg⁻¹. Soils were then incubated under unsaturated (50% of water holding capacity at saturation, WHC) or saturated (100% of WHC) water conditions for 7 and 36 d, respectively. During 7 d incubation of unsaturated soil, the NH₄⁺-N concentration decreased from 164.8 to 34.4 mg kg⁻¹, and the δ¹⁵N of NH₄⁺ increased from −0.4 to +57.2‰ through nitrification, as evidenced by corresponding increase in NO₃⁻-N concentration and lower δ¹⁵N of NO₃⁻ (product) than that of NH₄⁺ (substrate) at each sampling time. In saturated soil, the concentration of NH₄⁺-N decreased gradually from 162.4 to 24.2 mg kg⁻¹, and the δ¹⁵N values increased from +0.8 to +21.0‰ during 36 d incubation. However, increase in NO₃⁻ concentration was not observed due to loss of NO₃⁻ through concurrent denitrification in anaerobic sites. The apparent isotopic fractionation factors (α_s/p) associated with decrease in NH₄⁺ concentration were 1.04 and 1.01 in unsaturated and saturated soils, respectively. Since nitrification is likely to introduce greater isotope fractionation than microbial immobilization, the higher value for unsaturated soil probably reflected faster nitrification under aerobic conditions. The lower value for saturated soil suggests that immobilization and subsequent remineralization of NH₄⁺ were relatively more dominant than nitrification under the anaerobic conditions.     

N2O and NO emissions from an Andisol field as influenced by pelleted poultry manure

Animal manures from intensive livestock operations can be pelleted to improve handlings and recyclings of embodied nutrients. The aim of this study was to evaluate the influence of pelleted poultry manure on N₂O and NO fluxes from an Andisol field. In autumn 2006 and summer 2007, poultry manure (PM), pelleted poultry manure (PP), and chemical fertilizer (CF) were applied at a rate of 120 kg N ha⁻¹ in each cultivation period to Komatsuna (Brassica rapa var. peruviridis). Nitrous oxide and NO fluxes were measured using an automated monitoring system. A soil incubation experiment was also conducted to determine the influence of intact and ground pelleted manure on N₂O, NO, and CO₂ production with a water-filled pore space (WFPS) of 30 or 50%. In the field measurements, N₂O emission rates from the organic fertilizer treatments were larger than that from the CF treatment, possibly because organic C stimulated denitrification. The highest N₂O flux was observed from the PP treatment after a rainfall following fertilization, and the cumulative emission rate (2.72 ± 0.22 kg N ha⁻¹ y⁻¹) was 3.9 and 7.1 times that from the PM and CF treatments, respectively. In contrast, NO emission rates were highest from the CF treatment. The NO/N₂O flux ratio indicated that nitrification was the dominant process for NO and N₂O production from the CF treatment. Cumulative N₂O emission rates from all treatments were generally higher during the wetter cultivation period (autumn 2006) than during the drier cultivation period (summer 2007). In contrast, NO emission rates were higher in the drier than in the wetter cultivation period. The incubation experiment results showed a synergistic effect of soil moisture and the pelleted manure form on N₂O emission rates. The intact pelleted manure with the 50% WFPS treatment produced the highest N₂O and CO₂ fluxes and resulted in the lowest soil NO₃⁻ content after the incubation. These results indicate that anaerobic conditions inside the pellets, caused by rainfall and heterotrophic microbial activities, led to denitrification, resulting in high N₂O fluxes. Controlling the timing of N application by avoiding wet conditions might be one mitigation option to reduce N₂O emission rates from the PP treatment in this study field.     

How important is N2O production in removing atmospherically deposited nitrogen from UK moorland catchments?

Nitrate (NO₃⁻) leaching due to anthropogenic nitrogen (N) deposition is an environmental problem in many parts of the UK uplands, associated with surface water acidification and affecting lake nutrient balances. It is often assumed that gaseous return of deposited N to the atmosphere as N₂O through denitrification may provide an important sink for N. This assumption was tested for four moorland catchments (Allt a’Mharcaidh in the Cairngorms, Afon Gwy in mid-Wales, Scoat Tarn in the English Lake District and River Etherow in the southern Pennines), covering gradients of atmospheric N deposition and surface water NO₃⁻ leaching, through a combination of field and laboratory experiments. Field measurements of N₂O fluxes from static chambers with and without additions of NH₄NO₃ solution were carried out every 4 weeks over 1 yr. Wetted soil cores from the same field plots were used in experimental laboratory incubations at 5 and 15 °C with and without additions of NH₄NO₃ solution, followed by measurement of N₂O fluxes. Field measurements showed that significant N₂O fluxes occurred in only a very small number of plots with most showing zero values for much of the year. The maximum fluxes were 0.24 kg-N/ha/yr from unamended plots at the River Etherow and 0.49 kg-N/ha/yr from plots with NH₄NO₃ additions at the Allt a’Mharcaidh. Laboratory incubation experiments demonstrated that large N₂O fluxes could be induced by warming and NH₄NO₃ additions, with the top 5 cm of soil cores responsible for the largest fluxes, reaching 11.8 kg-N/ha/yr from a podsol at Scoat Tarn. Acetylene block experiments showed that while N₂ was not likely to be a significant denitrification product in these soils, reduced N₂O fluxes indicated that nitrification was an important source of N₂O in many cases. A simple model of denitrification suggesting that 10-80% of net N inputs may be denitrified from non-agricultural soils was found to greatly over-estimate fluxes in the UK uplands. The proportion of deposition denitrified was found to be much closer to the IPCC suggested value of 1% with an upper limit of 10%. Interception of N deposition by vegetation may greatly reduce the net supply of N from this source, while soil acidification or other factors limiting carbon supply to soil microbes may prevent large denitrification fluxes even where NO₃⁻ supply is not limiting.     

Nitrogen oxides emission from soils bearing a potato crop as influenced by fertilization with treated pig slurries and composts

Nitrous oxide, nitric oxide and denitrification losses from an irrigated soil amended with organic fertilizers with different soluble organic carbon fractions and ammonium contents were studied in a field study covering the growing season of potato (Solanum tuberosum). Untreated pig slurry (IPS) with and without the nitrification inhibitor dicyandiamide (DCD), digested thin fraction of pig slurry (DTP), composted solid fraction of pig slurry (CP) and composted municipal solid waste (MSW) mixed with urea were applied at a rate of 175 kg available N ha⁻¹, and emissions were compared with those from urea (U) and a control treatment without any added N fertilizer (Control). The cumulative denitrification losses correlated significantly with the soluble carbohydrates, dissolved N and total C added. Added dissolved organic C (DOC) and dissolved N affected the N₂O/N₂ ratio, and a lower ratio was observed for organic fertilizers than from urea or unfertilized controls. The proportion of N₂O produced from nitrification was higher from urea than from organic fertilizers. Accumulated N₂O losses during the crop season ranged from 3.69 to 7.31 kg N₂O-N ha⁻¹ for control and urea, respectively, whereas NO losses ranged from 0.005 to 0.24 kg NO-N ha⁻¹, respectively. Digested thin fraction of pig slurry compared to IPS mitigated the total N₂O emission by 48% and the denitrification rate by 33%, but did not influence NO emissions. Composted pig slurry compared to untreated pig slurry increased the N₂O emission by 40% and NO emission by 55%, but reduced the denitrification losses (34%). DCD partially inhibited nitrification rates and reduced N₂O and NO emissions from pig slurry by at least 83% and 77%, respectively. MSW+U, with a C:N ratio higher than that of the composted pig slurry, produced the largest denitrification losses (33.3 kg N ha⁻¹), although N₂O and NO emissions were lower than for the U and CP treatments.This work has shown that for an irrigated clay loam soil additions of treated organic fertilizers can mitigate the emissions of the atmospheric pollutants NO and N₂O in comparison with urea.     

Denitrification in the saturated zone of hydromorphic soils—laboratory measurement, regulating factors and stochastic modeling

Denitrification capacity (D_cap) was measured in 595 samples from the saturated zone of 18 hydromorphic soils by anaerobic incubation of K¹⁵NO₃ amended slurries in the laboratory and analyzing ¹⁵N₂+¹⁵N₂O in the sample headspace. Furthermore, texture, pH, C (total, dissolved, hot-water soluble organic and inorganic C), total N and sulfide were measured in order to model D_cap by multiple regression analysis. D_cap was significantly correlated with most properties, but exhibited closest relationships to electron donors (C_org, sulfide). Mean D_cap of the soil material groups sand, loam and peat was 0.13, 1.34 and 26.6 mg N kg⁻¹ d⁻¹. Regression models for predicting D_cap were varied in the selection of independent variables and in the differentiation of the source data-set. The models were found to be suitable to estimate mean D_cap of sites with medium or high activity. Best fits were obtained with the most complex models, but prediction for individual samples and for means of sites with low activity was not satisfactory with any of the models. The model based on organic C and total N and differentiating by type of soil material and position above or below the groundwater level was validated using an independent data-set of 46 samples from 10 sites of fluvial, glaciofluvial or glacial till sediments. The fluvial sediments exhibited satisfactory agreement between measured and predicted D_cap, whereas the other sites were overestimated because D_cap was too low and possibly because glacial till was not part of the source data-set. Our study provides for the first time statistical models of denitrification in shallow groundwater which are transferable to other sites with certain restrictions and which can thus be used for environmental modeling and soil mapping.     

Nitrogen fertilizers promote denitrification

A laboratory study was conducted to compare the effects of different N fertilizers on emission of N₂ and N₂O during denitrification of NO₃ ^– in waterlogged soil. Field-moist samples of Drummer silty clay loam soil (fine-silty, mixed, mesic Typic Haplaquoll) were incubated under aerobic conditions for 0, 2, 4, 7, 14, 21, or 42 days with or without addition of unlabelled (NH₄)₂SO₄, urea, NH₄H₂PO₄, (NH₄)₂HPO₄, NH₄NO₃ (200 or 1000 mg N kg^–1 soil), or liquid anhydrous NH₃ (1000 mg N kg^–1 soil). The incubated soil samples were then treated with ¹⁵N-labelled KNO₃ (250 mg N kg^–1 soil, 73.7 atom% ¹⁵N), and incubation was carried out under waterlogged conditions for 5 days, followed by collection of atmospheric samples for ¹⁵N analyses to determine labelled N₂ and N₂O. Compared to samples incubated without addition of unlabelled N, all of the fertilizers promoted denitrification of ¹⁵NO₃ ^–. Emission of labelled N₂ and N₂O decreased in the order: Anhydrous NH₃>urea<$>\gg<$> (NH₄)₂HPO₄>(NH₄)₂SO₄≃NH₄NO₃≃NH₄H₂PO₄. The highest emissions observed with anhydrous NH₃ or urea coincided with the presence of NO₂ ^–, and ¹⁵N analyses indicated that these emissions originated from NO₂ ^– rather than NO₃ ^–. Emissions of labelled N₂ and N₂O were significantly correlated with fertilizer effects on soil pH and water-soluble organic C. Received: 17 January 1996