Variations in soil N cycling and trace gas emissions in wet tropical forests

We used a previously described precipitation gradient in a tropical montane ecosystem of Hawai’i to evaluate how changes in mean annual precipitation (MAP) affect the processes resulting in the loss of N via trace gases. We evaluated three Hawaiian forests ranging from 2200 to 4050 mm year⁻¹ MAP with constant temperature, parent material, ecosystem age, and vegetation. In situ fluxes of N₂O and NO, soil inorganic nitrogen pools (NH₄⁺ and NO₃⁻), net nitrification, and net mineralization were quantified four times over 2 years. In addition, we performed ¹⁵N-labeling experiments to partition sources of N₂O between nitrification and denitrification, along with assays of nitrification potential and denitrification enzyme activity (DEA). Mean NO and N₂O emissions were highest at the mesic end of the gradient (8.7±4.6 and 1.1±0.3 ng N cm⁻² h⁻¹, respectively) and total oxidized N emitted decreased with increased MAP. At the wettest site, mean trace gas fluxes were at or below detection limit (≤0.2 ng N cm⁻² h⁻¹). Isotopic labeling showed that with increasing MAP, the source of N₂O changed from predominately nitrification to predominately denitrification. There was an increase in extractible NH₄⁺ and decline in NO₃⁻, while mean net mineralization and nitrification did not change from the mesic to intermediate sites but decreased dramatically at the wettest site. Nitrification potential and DEA were highest at the mesic site and lowest at the wet site. MAP exerts strong control N cycling processes and the magnitude and source of N trace gas flux from soil through soil redox conditions and the supply of electron donors and acceptors.     

Natural 15N abundance of soil N pools and N2O reflect the nitrogen dynamics of forest soils

Natural ¹⁵N abundance measurements of ecosystem nitrogen (N) pools and ¹⁵N pool dilution assays of gross N transformation rates were applied to investigate the potential of δ¹⁵N signatures of soil N pools to reflect the dynamics in the forest soil N cycle. Intact soil cores were collected from pure spruce (Picea abies (L.) Karst.) and mixed spruce-beech (Fagus sylvatica L.) stands on stagnic gleysol in Austria. Soil δ¹⁵N values of both forest sites increased with depth to 50 cm, but then decreased below this zone. δ¹⁵N values of microbial biomass (mixed stand: 4.7 ± 0.8‰, spruce stand: 5.9 ± 0.9‰) and of dissolved organic N (DON; mixed stand: 5.3 ± 1.7‰, spruce stand: 2.6 ± 3.3‰) were not significantly different; these pools were most enriched in ¹⁵N of all soil N pools. Denitrification represented the main N₂O-producing process in the mixed forest stand as we detected a significant ¹⁵N enrichment of its substrate NO₃⁻ (3.6 ± 4.5‰) compared to NH₄⁺ (−4.6 ± 2.6‰) and its product N₂O (−11.8 ± 3.2‰). In a ¹⁵N-labelling experiment in the spruce stand, nitrification contributed more to N₂O production than denitrification. Moreover, in natural abundance measurements the NH₄⁺ pool was slightly ¹⁵N-enriched (−0.4 ± 2.0 ‰) compared to NO₃⁻ (−3.0 ± 0.6 ‰) and N₂O (−2.1 ± 1.1 ‰) in the spruce stand, indicating nitrification and denitrification operated in parallel to produce N₂O. The more positive δ¹⁵N values of N₂O in the spruce stand than in the mixed stand point to extensive microbial N₂O reduction in the spruce stand. Combining natural ¹⁵N abundance and ¹⁵N tracer experiments provided a more complete picture of soil N dynamics than possible with either measurement done separately.     

Modeling nitrogen removal for a denitrification biofilter

Nitrous oxide (N₂O) is a major greenhouse gas, heavily contributing to global warming. N₂O is emitted from various sources such as wastewater treatment plants, during the nitrification and denitrification steps. ASM models, which are commonly used in wastewater treatment, usually consider denitrification as a one-step process (NO₃ ⁻ directly reduced to N₂) and are as such unable to provide values for intermediate products of the reaction like N₂O. In this study, a slightly modified ASM1 model was implemented in the GPS-X™ software to simulate the concentration of such intermediate products (NO₂ ⁻, NO and N₂O) and to estimate the amounts of gaseous N₂O emitted by the denitrification stage (12 biofilters) of the Seine-Centre WWTP (SIAAP, Paris). Simulations running on a 1-year period have shown good agreements with measured effluent data for nitrate and nitrite. The calculated mean value for emitted N₂O is 4.95 kgN–N₂O/day, which stands in the typical range of estimated experimental values of 4–31 kgN–N₂O/day. Nitrous oxide emissions are usually not measured on WWTPs and so, as obtained results show, there is a certain potential for using models that quantify those emissions using traditionally measured influent data.     

Spatial and Temporal Variability in Sediment Denitrification Within an Agriculturally Influenced Reservoir

Reservoirs are intrinsically linked to the rivers that feed them, creating a river–reservoir continuum in which water and sediment inputs are a function of the surrounding watershed land use. We examined the spatial and temporal variability of sediment denitrification rates by sampling longitudinally along an agriculturally influenced river–reservoir continuum monthly for 13 months. Sediment denitrification rates ranged from 0 to 63 μg N₂O g ash free dry mass of sediments (AFDM)⁻¹ h⁻¹ or 0–2.7 μg N₂O g dry mass of sediments (DM)⁻¹ h⁻¹ at reservoir sites, vs. 0–12 μg N₂O gAFDM⁻¹ h⁻¹ or 0–0.27 μg N₂O gDM⁻¹ h⁻¹ at riverine sites. Temporally, highest denitrification activity traveled through the reservoir from upper reservoir sites to the dam, following the load of high nitrate (NO₃⁻-N) water associated with spring runoff. Annual mean sediment denitrification rates at different reservoir sites were consistently higher than at riverine sites, yet significant relationships among theses sites differed when denitrification rates were expressed per gDM vs. per gAFDM. There was a significant positive relationship between sediment denitrification rates and NO₃⁻-N concentration up to a threshold of 0.88 mg NO₃⁻ -N l⁻¹, above which it appeared NO₃⁻-N was no longer limiting. Denitrification assays were amended seasonally with NO₃⁻-N and an organic carbon source (glucose) to determine nutrient limitation of sediment denitrification. While organic carbon never limited sediment denitrification, all sites were significantly limited by NO₃⁻-N during fall and winter when ambient NO ₃⁻-N was low.     

Nitrous oxide in brackish Lakes Shinji and Nakaumi, Japan

Nitrous oxide (N₂O) was measured monthly from September 1997 to August 1998 in the brackish Lakes Shinji and Nakaumi, Japan. N₂O (5–37 μg N l⁻¹) was supersaturated in the overlying water on lake sediments from October 1997 to January 1998. The N₂O concentration in the hypolimnion was higher than that in the epilimnion on 17 October 1997, when N₂O was first observed in a water column of Lake Nakaumi. Afterward, N₂O was almost uniform throughout the water column and then disappeared on 16 February 1998. On the one hand, large amounts of N₂O were found throughout the year in the interstitial water in Lake Shinji, where a high concentration of nitrate was discharged from the Hii River. On the other hand, in Lake Nakaumi, stratified by halocline, a high concentration of N₂O was observed in the interstitial water only from winter to spring. N₂O concentrations in the interstitial water were about 10 to 1000 times as large as those in the overlying water. These results imply that N₂O was mainly produced at the sediment-water interface and was diffused to the overlying water. It was also suggested that the accumulation of N₂O in the sediment-water system was accelerated by a high concentration of hydrogen sulfide. Received: July 6, 2000 / Accepted: November 30, 2000     

Ecosystem metabolism and greenhouse gas production in a mesotrophic northern temperate lake experiencing seasonal hypoxia

Many lacustrine systems, despite management efforts to control eutrophication, are hypoxic during stratified periods. Hypoxia is a major concern, not only for its impact on aquatic life but also for its potential to stimulate production of the greenhouse gases, methane (CH₄) and nitrous oxide (N₂O). We investigated the drivers of hypoxia in Muskegon Lake, a temperate dimictic freshwater estuary that experiences frequent hypolimnetic mixing due to atmospheric forces, riverine inputs, and intrusion of oxic water from coastal upwelling in Lake Michigan. Primary production and respiration (R) rates obtained from a δ¹⁸O mass balance model were similar to other mesotrophic environments (0.56–26.31 and 0.57–13.15 mmol O₂ m^?3 day^?1, respectively), although high P/R (≥2 in mid-summer) indicated there is sufficient autochthonous production to support hypoxia development and persistence. The isotopic enrichment factor for respiration (ε_obs) varied markedly and was least negative in August of both sampling years, consistent with high R rates. Hypoxic conditions were associated with accumulation of N₂O but not CH₄, and emissions of N₂O are among the highest reported from lakes. The average N₂O site preference value of 25.4‰ indicates that the majority of N₂O was produced by nitrification via hydroxylamine oxidation, despite the presence of resilient hypoxia. While it has been hypothesized that denitrification acts as a sink for N₂O in hypoxic lakes, it is clear that Muskegon Lake functions as a strong source of N₂O via nitrification. Further considerations of lakes as global sources of N₂O thus warrant a closer evaluation of nitrification-fueled N₂O production.     

Biosensor Determination of the Microscale Distribution of Nitrate, Nitrate Assimilation, Nitrification, and Denitrification in a Diatom-Inhabited Freshwater Sediment

High-resolution NO₃⁻ profiles in freshwater sediment covered with benthic diatoms were obtained with a new microscale NO₃⁻ biosensor characterized by absence of interference from chemical species other than NO₂⁻ and N₂O. Analysis of the microprofiles obtained indicated no nitrification during darkness, high rates of nitrification and a tight coupling between nitrification and denitrification during illumination, and substantial rates of NO₃⁻ assimilation during illumination. Nitrification during darkness could be induced by purging the bulk water with O₂ gas, indicating that the stimulatory effect on nitrification by illumination was caused by algal production of O₂. NH₄⁺ addition did not stimulate nitrification during darkness when O₂ was restricted to the upper 1-mm layer, and there was thus a low nitrification potential in the permanently oxic top 1 mm of the sediment.     

Nitrogen removal in a wastewater treatment plant through biofilters: nitrous oxide emissions during nitrification and denitrification

In order to estimate N₂O emissions from immersed biofilters during nitrogen removal in tertiary treatments at urban wastewater treatment plants (WWTPs), a fixed culture from the WWTP of “Seine Centre” (Paris conurbation) was subjected to lab-scale batch experiments under various conditions of oxygenation and a gradient of methanol addition. The results show that during nitrification, N₂O emissions are positively related to oxygenation (R ² = 0.99). However, compared to the rates of ammonium oxidation, the percentage of emitted N₂O is greater when oxygenation is low (0.5–1 mgO₂ L⁻¹), representing up to 1% of the oxidized ammonium (0.4% on average). During denitrification, the N₂O emission reaches a significant peak when the quantity of methanol allows denitrification of between 66% and 88%. When methanol concentrations lead to a denitrification of close to 100%, the flows of N₂O are much lower and represent on average 0.2% of the reduced nitrate. By considering these results, we can estimate, the emissions of N₂O during nitrogen removal, at the “Seine Centre” WWTP, to approximately 38 kgN-N₂O day⁻¹.     

Distinguishing between Nitrification and Denitrification as Sources of Gaseous Nitrogen Production in Soil

The source of N₂O produced in soil is often uncertain because denitrification and nitrification can occur simultaneously in the same soil aggregate. A technique which exploits the differential sensitivity of these processes to C₂H₂ inhibition is proposed for distinguishing among gaseous N losses from soils. Denitrification N₂O was estimated from 24-h laboratory incubations in which nitrification was inhibited by 10-Pa C₂H₂. Nitrification N₂O was estimated from the difference between N₂O production under no C₂H₂ and that determined for denitrification. Denitrification N₂ was estimated from the difference between N₂O production under 10-kPa C₂H₂ and that under 10 Pa. Laboratory estimates of N₂O production were significantly correlated with in situ N₂O diffusion measurements made during a 10-month period in two forested watersheds. Nitrous oxide production from nitrification was most important on well-drained sites of a disturbed watershed where ambient NO₃⁻ was high. In contrast, denitrification N₂O was most important on poorly drained sites near the stream of the same watershed. Distinction between N₂O production from nitrification and denitrification was corroborated by correlations between denitrification N₂O and water-filled pore space and between nitrification N₂O and ambient NO₃⁻. This technique permits qualitative study of environmental parameters that regulate gaseous N losses via denitrification and nitrification.     

Microbial N Turnover and N-Oxide (N<Subscript>2</Subscript>O/NO/NO<Subscript>2</Subscript>) Fluxes in Semi-arid Grassland of Inner Mongolia

Gross rates of N mineralization and nitrification, and soil–atmosphere fluxes of N₂O, NO and NO₂ were measured at differently grazed and ungrazed steppe grassland sites in the Xilin river catchment, Inner Mongolia, P. R. China, during the 2004 and 2005 growing season. The experimental sites were a plot ungrazed since 1979 (UG79), a plot ungrazed since 1999 (UG99), a plot moderately grazed in winter (WG), and an overgrazed plot (OG), all in close vicinity to each other. Gross rates of N mineralization and nitrification determined at in situ soil moisture and soil temperature conditions were in a range of 0.5–4.1 mg N kg⁻¹ soil dry weight day⁻¹. In 2005, gross N turnover rates were significantly higher at the UG79 plot than at the UG99 plot, which in turn had significantly higher gross N turnover rates than the WG and OG plots. The WG and the OG plot were not significantly different in gross ammonification and in gross nitrification rates. Site differences in SOC content, bulk density and texture could explain only less than 15% of the observed site differences in gross N turnover rates. N₂O and NO _x flux rates were very low during both growing seasons. No significant differences in N trace gas fluxes were found between plots. Mean values of N₂O fluxes varied between 0.39 and 1.60 μg N₂O-N m⁻² h⁻¹, equivalent to 0.03–0.14 kg N₂O-N ha⁻¹ y⁻¹, and were considerably lower than previously reported for the same region. NO _x flux rates ranged between 0.16 and 0.48 μg NO _x -N m⁻² h⁻¹, equivalent to 0.01–0.04 kg NO _x -N ha⁻¹ y⁻¹, respectively. N₂O fluxes were significantly correlated with soil temperature and soil moisture. The correlations, however, explained only less than 20% of the flux variance.     

Denitrification, Dissimilatory Reduction of Nitrate to Ammonium, and Nitrification in a Bioturbated Estuarine Sediment as Measured with 15N and Microsensor Techniques

Nitrogen and oxygen transformations were studied in a bioturbated (reworked by animals) estuarine sediment (Norsminde Fjord, Denmark) by using a combination of ¹⁵N isotope (NO₃^-), specific inhibitor (C₂H₂), and microsensor (N₂O and O₂) techniques in a continuous-flow core system. The estuarine water was NO₃^- rich (125 to 600 μM), and NO₃^- was consistently taken up by the sediment on the four occasions studied. Total NO₃^- uptake (3.6 to 34.0 mmol of N m^-2 day^-1) corresponded closely to N₂ production (denitrification) during the experimental steady state, which indicated that dissimilatory, as well as assimilatory, NO₃^- reduction to NH₄⁺ was insignificant. When C₂H₂ was applied in the flow system, denitrification measured as N₂O production was often less (58 to 100%) than the NO₃^- uptake because of incomplete inhibition of N₂O reduction. The NO₃^- formed by nitrification and not immediately denitrified but released to the overlying water, uncoupled nitrification, was calculated both from ¹⁵NO₃^- dilution and from changes in NO₃^- uptake before and after C₂H₂ addition. These two approaches gave similar results, with rates ranging between 0 and 8.1 mmol of N m^-2 day^-1 on the four occasions. Attempts to measure total nitrification activity by the difference between NH₄⁺ fluxes before and after C₂H₂ addition failed because of non-steady-state NH₄⁺ fluxes. The vertical distribution of denitrification and oxygen consumption was studied by use of N₂O and O₂ microelectrodes. The N₂O profiles measured during the experimental steady state were often irregularly shaped, and the buildup of N₂O after C₂H₂ was added was much too fast to be described by a simple diffusion model. Only bioturbation by a dense population of infauna could explain these observations. This was corroborated by the relationship between diffusive and total fluxes, which showed that only 19 to 36 and 29 to 62% of the total O₂ uptake and denitrification, respectively, were due to diffusion-reaction processes at the regular sediment surface, excluding animal burrows.     

Temporal Change in Nitrous Oxide and Dinitrogen from Denitrification Following Onset of Anaerobiosis

Similar temporal patterns were found in three mineral soils for the composition of the gaseous products of denitrification following the onset of anaerobic conditions. During the early period of anaerobiosis (0 up to 1 to 3 h), N₂ was the dominant product of denitrification. The NO₃⁻ → N₂O activity then increased, but was not accompanied by a corresponding increase in N₂O-reducing activity. This resulted in a relatively extended period of time (1 to 3 up to 16 to 33 h) during which N₂O was a major product. Eventually (after 16 to 33 h), an increase in N₂O-reducing activity occurred without a comparable increase in the N₂O-producing activity. The increase in the rate of N₂O reduction did not occur in the presence of chloramphenicol and required the presence of N₂O or NO₃⁻ during the preceding anaerobic incubation. During the final period (16 to 33, up to 48 h), N₂ was generally the sole product of denitrification, since the rate of N₂O reduction exceeded the rate of N₂O production. A similar sequential pattern was also found for a culture of a denitrifying Flavobacterium sp. shifted to anaerobic growth. A staggered synthesis of the enzymes in the denitrification sequence apparently occurred in response to anoxia, which caused first a net production of N₂O followed by consumption of N₂O.     

Fungal control of nitrous oxide production in semiarid grassland

Fungi are capable of both nitrification and denitrification and dominate the microbial biomass in many soils. Recent work suggests that fungal rather than bacterial pathways dominate N transformation in desert soils. We evaluated this hypothesis by comparing the contributions of bacteria and fungi to N₂O production at control and N fertilized sites within a semiarid grassland in central New Mexico (USA). Soil samples were taken from the rhizosphere of blue grama (B. gracilus) and the microbiotic crusts that grow in open areas between the bunch grasses. Soils incubated at 30% or 70% water holding capacity, were exposed to one of three biocide treatments (control, cycloheximide or streptomycin). After 48 h, N₂O and CO₂ production were quantified along with the activities of several extracellular enzymes. N₂O production from N fertilized soils was higher than that of control soils (165 vs. 41 pmol h⁻¹ g⁻¹), was higher for crust soil than for rhizosphere soil (108 vs. 97 pmol h⁻¹ g⁻¹), and increased with soil water content (146 vs. 60 pmol h⁻¹ g⁻¹). On average, fungicide (cycloheximide) addition reduced N₂O production by 85% while increasing CO₂ production by 69%; bactericide (streptomycin) reduced N₂O by 53% with mixed effects on CO₂ production. N₂O production was significantly correlated with C and N mineralization potential as measured by assays for glycosidic and proteolytic enzymes, and with extractable nitrate and ammonium. Our data indicate that fungal nitrifier denitrification and bacterial autotrophic nitrification dominate N transformation in this ecosystem and that N₂O production is highly sensitive to soil cover, N deposition and moisture.     

Simulation Model of the Coupling between Nitrification and Denitrification in a Freshwater Sediment

A model was constructed to simulate the results of experiments which investigated nitrification and denitrification in the freshwater sediment of Lake Vilhelmsborg, Denmark (K. Jensen, N. P. Sloth, N. Risgaard-Petersen, S. Rysgaard, and N. P. Revsbech, Appl. Environ. Microbiol. 60:2094-2100, 1994). The model output faithfully represented the profiles of O₂ and NO₃^- and rates of nitrification, denitrification, and O₂ consumption as the O₂ concentration in the overlying water was increased from 10 to 600 μM. The model also accurately predicted the response, to increasing O₂ concentrations, of the integrated (micromoles per square meter per hour) rates of nitrification and denitrification. The simulated rates of denitrification of NO₃^- diffusing from the overlying water (D_w) and of NO₃^- generated by nitrification within the sediment (D_n) corresponded to the experimental rates as the O₂ concentration in the overlying water was altered. The predicted D_w and D_n rates, as NO₃^- concentration in the overlying water was changed, closely resembled those determined experimentally. The model was composed of 41 layers 0.1 mm thick, of which 3 represented the diffusive boundary layer in the water. Large first-order rate constants for nitrification and denitrification were required to completely oxidize all NH₄⁺ diffusing from the lower sediment layers and to remove much of the NO₃^- produced. In addition to the flux of NH₄⁺ from below, the model required a flux of an electron donor, possibly methane. Close coupling between nitrification and denitrification, achieved by allowing denitrification to tolerate some O₂ (~10 μM), was necessary to reproduce the real data. Spatial separation of the two processes (no toleration by denitrification of O₂) resulted in too high NO₃^- concentrations and too low rates of denitrification.     

Mechanism leading to N<Subscript>2</Subscript>O production in wastewater treating biofilm systems

Wastewater treatment plants are known to be important point sources for nitrous oxide (N₂O) in the anthropogenic N cycle. Biofilm based treatment systems have gained increasing popularity in the treatment of wastewater, but the mechanisms and controls of N₂O formation are not fully understood. Here, we review functional groups of microorganism involved in nitrogen (N) transformations during wastewater treatment, with emphasis on potential mechanism of N₂O production in biofilms. Biofilms used in wastewater treatment typically harbour aerobic and anaerobic zones, mediating close interactions between different groups of N transforming organisms. Current models of mass transfer and biomass interactions in biofilms are discussed to illustrate the complex regulation of N₂O production. Ammonia oxidizing bacteria (AOB) are the prime source for N₂O in aerobic zones, while heterotrophic denitrifiers dominate N₂O production in anoxic zones. Nitrosative stress ensuing from accumulation of NO₂ ^? during partial nitrification or denitrification seems to be one of the most critical factors for enhanced N₂O formation. In AOB, N₂O production is coupled to nitrifier denitrification triggered by nitrosative stress, low O₂ tension or low pH. Chemical N₂O production from AOB intermediates (NH₂OH, HNO, NO) released during high NH₃ turnover seems to be limited to surface-near AOB clusters, since diffusive mass transport resistance for O₂ slows down NH₃ oxidation rates in deeper biofilm layers. The proportion of N₂O among gaseous intermediates (NO, N₂O, N₂) in heterotrophic denitrification increases when NO or nitrous acid (HNO₂) accumulates because of increasing NO₂ ^?, or when transient oxygen intrusion impairs complete denitrification. Limited electron donor availability due to mass transport limitation of organic substrates into anoxic biofilm zones is another important factor supporting high N₂O/N₂ ratios in heterotrophic denitrifiers. Biofilms accommodating Anammox bacteria release less N₂O, because Anammox bacteria have no known N₂O producing metabolism and reduce NO₂ ^? to N₂, thereby lowering nitrosative stress to AOB and heterotrophs.     

Denitrification and N2O effluxes in the Bothnian Bay (northern Baltic Sea) river sediments as affected by temperature under different oxygen concentrations

Denitrification rates and nitrous oxide (N₂O) effluxes were measured at different temperatures and for different oxygen concentrations in the sediments of a eutrophied river entering the Bothnian Bay. The experiments were made in a laboratory microcosm with intact sediment samples. ¹⁵N-labelling was used to measure denitrification rates (Dw). The rates were measured at four temperatures (5, 10, 15 and 20°C) and with three oxygen inputs (<0.2, 5, and 10 mg O₂ l⁻¹). The temperature response was highly affected by oxygen concentration. At higher O₂ concentrations (5 and 10 mg O₂ l⁻¹) a saturation over 10°C was observed, whereas the anoxic treatment (<0.2 mg O₂ l⁻¹) showed an exponential increase in the temperature interval with a Q ₁₀ value of 3.1. The result is described with a combined statistical model. In contrast with overall denitrification, the N₂O effluxes from sediments decreased with increasing temperature. The N₂O effluxes had a lower response to oxygen than denitrification rates. The N₂O/N₂ ratio was always below 0.02. Increased temperatures in the future could enhance denitrification rates in boreal river sediments but would not increase the amount of N₂O produced.     

The saturation of N cycling in Central Plains streams: <Superscript>15</Superscript>N experiments across a broad gradient of nitrate concentrations

We conducted ¹⁵NO₃⁻ stable isotope tracer releases in nine streams with varied intensities and types of human impacts in the upstream watershed to measure nitrate (NO₃⁻) cycling dynamics. Mean ambient NO₃⁻ concentrations of the streams ranged from 0.9 to 21,000 μg l⁻¹ NO₃⁻–N. Major N-transforming processes, including uptake, nitrification, and denitrification, all increased approximately two to three orders of magnitude along the same gradient. Despite increases in transformation rates, the efficiency with which stream biota utilized available NO₃⁻-decreased along the gradient of increasing NO₃⁻. Observed functional relationships of biological N transformations (uptake and nitrification) with NO₃⁻ concentration did not support a 1st order model and did not show signs of Michaelis–Menten type saturation. The empirical relationship was best described by a Efficiency Loss model, in which log-transformed rates (uptake and nitrification) increase with log-transformed nitrate concentration with a slope less than one. Denitrification increased linearly across the gradient of NO₃⁻ concentrations, but only accounted for ∼1% of total NO₃⁻ uptake. On average, 20% of stream water NO₃⁻ was lost to denitrification per km, but the percentage removed in most streams was <5% km⁻¹. Although the rate of cycling was greater in streams with larger NO₃⁻ concentrations, the relative proportion of NO₃⁻ retained per unit length of stream decreased as NO₃⁻ concentration increased. Due to the rapid rate of NO₃⁻ turnover, these streams have a great potential for short-term retention of N from the landscape, but the ability to remove N through denitrification is highly variable.     

Role of nitrite reductase in the ammonia-oxidizing pathway of <Emphasis Type="Italic">Nitrosomonas europaea</Emphasis>

Metabolism of ammonia (NH₃) and hydroxylamine (NH₂OH) by wild-type and a nitrite reductase (nirK) deficient mutant of Nitrosomonas europaea was investigated to clarify the role of NirK in the NH₃ oxidation pathway. NirK-deficient N. europaea grew more slowly, consumed less NH₃, had a lower rate of nitrite (NO₂ ⁻) production, and a significantly higher rate of nitrous oxide (N₂O) production than the wild-type when incubated with NH₃ under high O₂ tension. In incubations with NH₃ under low O₂ tension, NirK-deficient N. europaea grew more slowly, but had only modest differences in NH₃ oxidation and product formation rates relative to the wild-type. In contrast, the nirK mutant oxidized NH₂OH to NO₂ ⁻ at consistently slower rates than the wild-type, especially under low O₂ tension, and lost a significant pool of NH₂OH–N to products other than NO₂ ⁻ and N₂O. The rate of N₂O production by the nirK mutant was ca. three times higher than the wild-type during hydrazine-dependent NO₂ ⁻ reduction under both high and low O₂ tension. Together, the results indicate that NirK activity supports growth of N. europaea by supporting the oxidation of NH₃ to NO₂ ⁻ via NH₂OH, and stimulation of hydrazine-dependent NO₂ ⁻ reduction by NirK-deficient N. europaea indicated the presence of an alternative, enzymatic pathway for N₂O production.     

Rapid Nitrate Loss and Denitrification in a Temperate River Floodplain

Nitrogen (N) pollution is a problem in many large temperate zone rivers, and N retention in river channels is often small in these systems. To determine the potential for floodplains to act as N sinks during overbank flooding, we combined monitoring, denitrification assays, and experimental nitrate (NO₃⁻ -N) additions to determine how the amount and form of N changed during flooding and the processes responsible for these changes in the Wisconsin River floodplain (USA). Spring flooding increased N concentrations in the floodplain to levels equal to the river. As discharge declined and connectivity between the river and floodplain was disrupted, total dissolved N decreased over 75% from 1.41 mg l⁻¹, equivalent to source water in the Wisconsin River on 14 April 2001, to 0.34 mg l⁻¹ on 22 April 2001. Simultaneously NO₃⁻ -N was attenuated almost 100% from 1.09 to <0.002 mg l⁻¹. Unamended sediment denitrification rates were moderate (0–483 μg m⁻² h⁻¹) and seasonally variable, and activity was limited by the availability of NO ₃⁻ -N on all dates. Two experimental NO₃⁻ -N pulse additions to floodplain water bodies confirmed rapid NO₃⁻ -N depletion. Over 80% of the observed NO ₃⁻ -N decline was caused by hydrologic export for addition #1 but only 22% in addition #2. During the second addition, a significant fraction (>60%) of NO₃⁻ -N mass loss was not attributable to hydrologic losses or conversion to other forms of N, suggesting that denitrification was likely responsible for most of the NO₃⁻ -N disappearance. Floodplain capacity to decrease the dominant fraction of river borne N within days of inundation demonstrates that the Wisconsin River floodplain was an active N sink, that denitrification often drives N losses, and that enhancing connections between rivers and their floodplains may enhance overall retention and reduce N exports from large basins.     

Denitrification and nitrous oxide emissions from riparian forests soils exposed to prolonged nitrogen runoff

Compared to upland forests, riparian forest soils have greater potential to remove nitrate (NO₃) from agricultural runoff through denitrification. It is unclear, however, whether prolonged exposure of riparian soils to nitrogen (N) loading will affect the rate of denitrification and its end products. This research assesses the rate of denitrification and nitrous oxide (N₂O) emissions from riparian forest soils exposed to prolonged nutrient runoff from plant nurseries and compares these to similar forest soils not exposed to nutrient runoff. Nursery runoff also contains high levels of phosphate (PO₄). Since there are conflicting reports on the impact of PO₄ on the activity of denitrifying microbes, the impact of PO₄ on such activity was also investigated. Bulk and intact soil cores were collected from N-exposed and non-exposed forests to determine denitrification and N₂O emission rates, whereas denitrification potential was determined using soil slurries. Compared to the non-amended treatment, denitrification rate increased 2.7- and 3.4-fold when soil cores collected from both N-exposed and non-exposed sites were amended with 30 and 60 μg NO₃-N g⁻¹ soil, respectively. Net N₂O emissions were 1.5 and 1.7 times higher from the N-exposed sites compared to the non-exposed sites at 30 and 60 μg NO₃-N g⁻¹ soil amendment rates, respectively. Similarly, denitrification potential increased 17 times in response to addition of 15 μg NO₃-N g⁻¹ in soil slurries. The addition of PO₄ (5 μg PO₄-P g⁻¹) to soil slurries and intact cores did not affect denitrification rates. These observations suggest that prolonged N loading did not affect the denitrification potential of the riparian forest soils; however, it did result in higher N₂O emissions compared to emission rates from non-exposed forest soils.