Measured Denitrification Rates Research Articles

Autonomous observations of biogenic N2 in the Eastern Tropical North Pacific using profiling floats equipped with gas tension devices

Oxygen Deficient Zones (ODZs) of the world’s oceans represent a relatively small fraction of the ocean by volume (<0.05% for suboxic and<5% for hypoxic) yet are receiving increased attention by experimentalists and modelers due to their importance in ocean nutrient cycling and predicted susceptibility to expansion and/or contraction forced by global warming. Conventional methods to study these biogeochemically important regions of the ocean have relied on well-developed but still relatively high cost and labor-intensive shipboard methods that include mass-spectrometric analysis of nitrogen-to-argon ratios (N2/Ar) and nutrient stoichiometry (relative abundance of nitrate, nitrite, and phosphate). Experimental studies of denitrification rates and processes typically involve either in-situ or in-vitro incubations using isotopically labeled nutrients. Over the last several years we have been developing a Gas Tension Device (GTD) to study ODZ denitrification including deployment in the largest ODZ, the Eastern Tropical North Pacific (ETNP). The GTD measures total dissolved gas pressure from which dissolved N2 concentration is calculated. Data from two cruises passing through the core of the ETNP near 17 °N in late 2020 and 2021 are presented, with additional comparisons at 12 °N for GTDs mounted on a rosette/CTD as well as modified profiling Argo-style floats. Gas tension was measured on the float with an accuracy of< 0.1% and relatively low precision (< 0.12%) when shallow (P< 200 dbar) and high precision (< 0.03%) when deep (P > 300 dbar). We discriminate biologically produced N2 (ie., denitrification) from N2 in excess of saturation due to physical processes (e.g., mixing) using a new tracer – ‘preformed excess-N2’. We used inert dissolved argon (Ar) to help test the assumption that preformed excess-N2 is indeed conservative. We used the shipboard measurements to quantify preformed excess-N2 by cross-calibrating the gas tension method to the nutrient-deficit method. At 17 °N preformed excess-N2 decreased from approximately 28 to 12 µmol/kg over σ0 = 24–27 kg/m3 with a resulting precision of ±1 µmol N2/kg; at 12 °N values were similar except in the potential density range of 25.7< σ0< 26.3 where they were lower by 1 µmol N2/kg due likely to being composed of different source waters. We then applied these results to gas tension and O2 (< 3 µmol O2/kg) profiles measured by the nearby float to obtain the first autonomous biogenic N2 profile in the open ocean with an RMSE of ± 0.78 µM N2, or ± 19%. We also assessed the potential of the method to measure denitrification rates directly from the accumulation of biogenic N2 during the float drifts between profiling. The results suggest biogenic N2 rates of ±20 nM N2/day could be detected over >16 days (positive rates would indicate denitrification processes whereas negative rates would indicate predominantly dilution by mixing). These new observations demonstrate the potential of the gas tension method to determine biogenic N2 accurately and precisely in future studies of ODZs.

Nitrogen loss processes in response to upwelling in a Peruvian coastal setting dominated by denitrification – a mesocosm approach

Abstract. Upwelling of nutrient-rich deep waters make eastern boundary upwelling systems (EBUSs), such as the Humboldt Current system, hot spots of marine productivity. Associated settling of organic matter to depth and consecutive aerobic decomposition results in large subsurface water volumes being oxygen depleted. Under these circumstances, organic matter remineralisation can continue via denitrification, which represents a major loss pathway for bioavailable nitrogen. Additionally, anaerobic ammonium oxidation can remove significant amounts of nitrogen in these areas. Here we assess the interplay of suboxic water upwelling and nitrogen cycling in a manipulative offshore mesocosm experiment. Measured denitrification rates in incubations with water from the oxygen-depleted bottom layer of the mesocosms (via 15N label incubations) mostly ranged between 5.5 and 20 nmol N2 L−1 h−1 (interquartile range), reaching up to 80 nmol N2 L−1 h−1. However, actual in situ rates in the mesocosms, estimated via Michaelis–Menten kinetic scaling, did most likely not exceed 0.2–4.2 nmol N2 L−1 h−1 (interquartile range) due to substrate limitation. In the surrounding Pacific, measured denitrification rates were similar, although indications of substrate limitation were detected only once. In contrast, anammox (anaerobic ammonium oxidation) made only a minor contribution to the overall nitrogen loss when encountered in both the mesocosms and the Pacific Ocean. This was potentially related to organic matter C / N stoichiometry and/or process-specific oxygen and hydrogen sulfide sensitivities. Over the first 38 d of the experiment, total nitrogen loss calculated from in situ rates of denitrification and anammox was comparable to estimates from a full nitrogen budget in the mesocosms and ranged between ∼ 1 and 5.5 µmol N L−1. This represents up to ∼ 20 % of the initially bioavailable inorganic and organic nitrogen standing stocks. Interestingly, this loss is comparable to the total amount of particulate organic nitrogen that was exported into the sediment traps at the bottom of the mesocosms at about 20 m depth. Altogether, this suggests that a significant portion, if not the majority of nitrogen that could be exported to depth, is already lost, i.e. converted to N2 in a relatively shallow layer of the surface ocean, provided that there are oxygen-deficient conditions like those during coastal upwelling in our study. Published data for primary productivity and nitrogen loss in all EBUSs reinforce such conclusion.

Denitrification in urban river sediment and the contribution to total nitrogen reduction

Streams and rivers, especially in urban areas, are substantial sinks of bioavailable nitrogen owing to their hydrological connections with terrestrial systems and the intensity of inputs mediated by human activities. Denitrification can account for nitrogen removal in rivers, but its importance in urban rivers has been scarcely studied. This study chose 8 representative rivers in Shanghai, under different level of urbanization, and measured denitrification rates of sediments across a year. The results showed significant differences in temporal, spatial and vertical distributions (p < 0.05). Seasonally, the denitrification rate in spring and winter were lower than in autumn. Spatially, the higher denitrification rate of rivers in residential and industrial areas than that in suburban rivers, which dominated with cultivated land, was weakly related to dissolved oxygen (DO) (p < 0.05, r = −0.380) and ammonium nitrogen (NH4+-N) concentration (p < 0.05, r = 0.303) in the water column. Vertically, the surface layer (0–2 cm) accounted for 78.2% of the total denitrification capacity (0–5 cm). The denitrification rates among different depths positively correlated with the extractable nitrate concentration (SNO3−-N), the extractable ammonium nitrogen concentration (SNH4+-N), the organic carbon concentration (SOC), water content and pH of sediment to varying degrees. In addition, the mean annual denitrification rate of studied rivers was calculated as 2.62 ± 0.420 mg N m−2 h−1, and denitrification contributed nearly 20% of the nitrogen removal, these results underscore the importance of denitrification in nitrogen reduction of urban rivers.

Contrasting subsurface denitrification characteristics under temperate pasture lands and its implications for nutrient management in agricultural catchments

Subsurface denitrification plays a key role in the reduction or ‘attenuation’ of nitrate contamination of groundwater and surface waters. We investigated subsurface denitrification characteristics in the vadose zone and shallow groundwater at four sites under pastoral farming in the Manawatū River catchment, located in the lower part of North Island, New Zealand. The denitrification potential of the vadose zone was determined by the laboratory incubation assays measuring the denitrifying enzyme activity (DEA) in soil samples collected from different soil horizons (up to 2.1 m below ground surface), whereas denitrification rates in shallow groundwaters were measured in situ by single-well, push-pull tests conducted in piezometers installed at multiple depths at the study sites. Soils and underlying geology, defining hydrogeologic settings, appear to influence the spatial variability of subsurface denitrification characteristics at the study sites. Where the vadose zone is thin and composed of coarse-textured soils, percolation of nitrate was evident in observed high nitrate-nitrogen concentrations (>5 mg L−1) in oxic and young shallow groundwaters, but low nitrate-nitrogen concentrations (<0.05 mg L−1) were observed in the reduced shallow groundwater found underneath the fine textured soils and/or a thick vadose zone. This was confirmed by the push-pull tests measuring denitrification rates from 0.08 to 1.07 mg N L−1 h−1 in the reduced shallow groundwaters (dissolved oxygen or DO < 0.5 mg L−1), while negligible in the oxic groundwaters (DO > 5 mg L−1) found at the study sites. These contrasting subsurface denitrification characteristics determine the ultimate delivery of nitrate losses from agricultural soils to receiving waters, where the fine textured thick vadose zone and reducing groundwater conditions offer nitrate reduction in the subsurface environment.

Tropical Coastal Wetlands Ameliorate Nitrogen Export During Floods

Wetlands can increase resilience to extreme climatic events and have a key role in protection and water quality improvement in coastal ecosystems. Studies in tropical coastal wetlands at a catchment scale are scarce, and most work has been undertaken on small, temperate wetlands. In this study, we tested whether natural coastal wetlands in a tropical catchment (Tully-Murray, Queensland, Australia) could ameliorate nitrogen (N) exported to the Great Barrier Reef during a flood event. We measured denitrification rates in different types of coastal wetlands (mangroves, saltmarshes, waterbodies with macrophytes, and floodplain wetlands dominated by Melaleuca spp.) to assess their potential contribution to N losses during the 6-day duration of a flood in March 2018. Denitrification potential was variable across the landscape, and we identified “hotspots” in sub-catchments with high NO−3-N concentrations (0.4–0.6 mg L−1) and large areas of wetlands (>800 ha, >40% of the sub-catchment). These hotspots can denitrify up to 10 t of NO−3-N per day during a flood. We used our measured denitrification rates to provide input parameters for a model that includes the main biogeochemical processes affecting N transformations within wetlands (nitrification, denitrification, plant uptake, sedimentation, anammox, and mineralization), and accounts for transport via the duration, depth, and flow of water. Model simulations of a sub-catchment of the Tully-Murray indicate that flood inundation of large areas of natural wetlands (>40% of the sub-catchment area) could potentially remove 70% of the incoming NO−3-N load in the first 24 h of the flood. The management and restoration of coastal tropical wetlands could play a critical role in sustaining the health of coastal ecosystems through water quality improvement.

Denitrification in the river network of a mixed land use watershed: unpacking the complexities

River networks have the potential to permanently remove nitrogen through denitrification. Few studies have measured denitrification rates within an entire river network or assessed how land use affect rates at larger spatial scales. We sampled 108 sites throughout the network of the Fox River watershed, Wisconsin, to determine if land use influence sediment denitrification rates, and to identify zones of elevated sediment denitrification rates (hot spots) within the river network. Partial least squares regression models identified variables from four levels of organization (river bed sediment, water column, riparian zone, and watershed) that best predicted denitrification rates throughout the river network. Nitrate availability was the most important predictor of denitrification rates, while land cover was not always a good predictor of local-scale nitrate concentrations. Thus, land cover and denitrification rate were not strongly related across the Fox River watershed. A direct relationship between denitrification rate and watershed land cover occurred only in the Wolf River sub-watershed, the least anthropogenically disturbed of the sub-watersheds. Denitrification hot spots were located throughout the river network, regardless of watershed land use, with hot spot location being determined primarily by nitrate availability. In the Fox River watershed, when nitrate was abundant, river bed sediment character influenced denitrification rate, with higher denitrification rates at sites with fine, organic sediments. These findings suggest that denitrification occurring throughout an entire river network, from headwater streams to larger rivers, can help reduce nitrogen loads to downstream water bodies.

Nitrogen removal by tropical floodplain wetlands through denitrification

Excess nitrogen (N) leading to the eutrophication of water and impacts on ecosystems is a serious environmental challenge. Wetlands can remove significant amounts of N from the water, primarily through the process of denitrification. Most of our knowledge on wetland denitrification is from temperate climates; studies in natural tropical wetlands are very scarce. We measured denitrification rates during a dry and a wet season in five floodplain forests dominated by Melaleuca spp., a coastal freshwater wetland of tropical Australia. We hypothesised that the denitrification potential of these wetlands would be high throughout the year and would be limited by N and carbon (C) availability. Mean potential denitrification rates (Dt) were 5.0±1.7mgm2h–1, and were within the reported ranges for other tropical and temperate wetlands. The rates of Dt were similar between the dry and the wet seasons. From the total unamended denitrification rates (Dw, 3.1±1.7mgm2h–1), 64% was derived from NO3– of the water column and the rest from coupled nitrification–denitrification. The factor most closely associated with denitrification was background water NO3–-N concentrations. Improved management and protection of wetlands could play an important role in improving water quality in tropical catchments.

Living shorelines enhance nitrogen removal capacity over time

Living shorelines are nature-based solutions to coastal erosion that can be constructed as salt marshes with fringing oyster reefs. Each of these habitats can decrease the potential for eutrophication through increased nitrogen (N) removal via denitrification. However, the development of N cycling over time has not been studied in living shorelines. This research measured denitrification rates in a chronosequence of living shorelines spanning 0–20 years in age in Bogue Sound, NC. Analyses were conducted seasonally from summer 2014 to spring 2015 along an elevation transect through the salt marsh, oyster reef, and adjacent sandflat at all sites. Gas fluxes (N2 and O2) from sediment core incubations were measured with a membrane inlet mass spectrometer (MIMS) to assess denitrification and sediment oxygen demand. Fluxes of dissolved nutrients and the greenhouse gas N2O were measured. Sediment properties, inundation frequency, oyster filtration rates, and marsh grass stem density were also quantified. There was no significant difference in denitrification rates among habitats. N removal consistently increased from the 0- to 7-year-old sites. Denitrification efficiency was always greater than 50% and positive N2O fluxes were negligible. Our results suggest that living shorelines increase net N removal within a relatively short time period following construction, without introducing deleterious greenhouse gas emissions. This demonstrates that living shorelines can play an important role in estuarine N cycling and management.

Watershed vs. within‐lake drivers of nitrogen: phosphorus dynamics in shallow lakes

Research on lake eutrophication often identifies variables affecting amounts of phosphorus (P) and nitrogen (N) in lakes, but understanding factors influencing N:P ratios is important given its influence on species composition and toxin production by cyanobacteria. We sampled 80 shallow lakes in Minnesota (USA) for three years to assess effects of watershed size, proportion of watershed as both row crop and natural area, fish biomass, and lake alternative state (turbid vs. clear) on total N:total P (TN : TP), ammonium, total dissolved phosphorus (TDP), and seston stoichiometry. We also examined N:P stoichiometry in 20 additional lakes that shifted states during the study. Last, we assessed the importance of denitrification by measuring denitrification rates in sediment cores from a subset of 34 lakes, and by measuring seston δ15 N in four additional experimental lakes before and after they were experimentally manipulated from turbid to clear states. Results showed alternative state had the largest influence on overall N:P stoichiometry in these systems, as it had the strongest relationship with TN : TP, seston C:N:P, ammonium, and TDP. Turbid lakes had higher N at given levels of P than clear lakes, with TN and ammonium 2-fold and 1.4-fold higher in turbid lakes, respectively. In lakes that shifted states, TN was 3-fold higher in turbid lakes, while TP was only 2-fold higher, supporting the notion N is more responsive to state shifts than is P. Seston δ15 N increased after lakes shifted to clear states, suggesting higher denitrification rates may be important for reducing N levels in clear states, and potential denitrification rates in sediment cores were among the highest recorded in the literature. Overall, our results indicate lake state was a primary driver of N:P dynamics in shallow lakes, and lakes in clear states had much lower N at a given level of P relative to turbid lakes, likely due to higher denitrification rates. Shallow lakes are often managed for the clear-water state due to increased value as wildlife habitat. However, our results indicate lake state also influences N biogeochemistry, such that managing shallow lakes for the clear-water state may also mitigate excess N levels at a landscape scale.

Buried particulate organic carbon stimulates denitrification and nitrate retention in stream sediments at the groundwater–surface water interface

AbstractThe interface between ground water and surface water in streams is a hotspot for N processing. However, the role of buried organic C in N transformation at this interface is not well understood, and inferences have been based largely on descriptive studies. Our main objective was to determine how buried particulate organic C (POC) affected denitrification and NO3− retention in the sediments of an upwelling reach in a sand-plains stream in Wisconsin. We manipulated POC in mesocosms inserted in the sediments. Treatments included low and high quantities of conditioned red maple leaves (buried beneath combusted sand), ambient sediment (sand containing background levels of POC), and a control (combusted sand). We measured denitrification rates in sediments by acetylene-block assays in the laboratory and by changes in N2 concentrations in the field using membrane inlet mass spectrometry. We measured NO3−, NH4+, and dissolved organic N (DON) retention as changes in concentrations and fluxes along groundw...

Soil denitrification fluxes from three northeastern North American forests across a range of nitrogen deposition.

In northern forests, large amounts of missing N that dominate N balances at scales ranging from small watersheds to large regional drainage basins may be related to N-gas production by soil microbes. We measured denitrification rates in forest soils in northeastern North America along a N deposition gradient to determine whether N-gas fluxes were a significant fate for atmospheric N inputs and whether denitrification rates were correlated with N availability, soil O2 status, or forest type. We quantified N2 and N2O fluxes in the laboratory with an intact-core method and monitored soil O2, temperature and moisture in three forests differing in natural and anthropogenic N enrichment: Turkey Lakes Watershed, Ontario; Hubbard Brook Experimental Forest, New Hampshire; and Bear Brook Watershed, Maine (fertilized and reference plots in hardwood and softwood stands). Total N-gas flux estimates ranged from <1 in fertilized hardwood uplands at Bear Brook to >100 kg N ha(-1) year(-1) in hardwood wetlands at Turkey Lakes. N-gas flux increased systematically with natural N enrichment from soils with high nitrification rates (Bear Brook < Hubbard Brook < Turkey Lakes) but did not increase in the site where N fertilizer has been added since 1989 (Bear Brook). Our results show that denitrification is an important and underestimated term (1-24% of atmospheric N inputs) in N budgets of upland forests in northeastern North America, but it does not appear to be an important sink for elevated anthropogenic atmospheric N deposition in this region.

Channel complexity and nitrate concentrations drive denitrification rates in urban restored and unrestored streams

Stream restoration projects constructed in urbanizing watersheds provided an opportunistic setting to determine how channel bedform influenced nitrate removal by denitrification. We measured denitrification rates in streambed sediments seasonally at eight streams in North Carolina, USA to characterize the physicochemical drivers of nitrogen transformations in restored urban streams. Mean denitrification rates were highly variable (97±58 to 585±214μmolm−2h−1) and most significantly influenced by nitrate concentrations; baseflow nitrate concentrations ranged from 0.06 to 1.07mg/L. A multiple linear regression approach also revealed the importance of channel complexity, water depth and temperature, which together with nitrate concentrations explained 60% of the variability among sites and seasons. Controlling for nutrient limitation, we observed significantly greater denitrification rates in more geomorphically complex streams, particularly near grade control structures and in deep pools. However, these trends were not consistently observed across all sites, which suggest the added influence of watershed scale drivers (e.g., urban hydrology) as well as reach scale drivers including the design, construction and age of the restoration project.

Insect emergence as a nitrogen flux in Neotropical streams: comparisons with microbial denitrification across a stream phosphorus gradient

AbstractResearch on how N is retained and removed from stream networks has focused on microbial metabolic pathways, such as denitrification. An alternative pathway for N to escape streams is in the form of emerging aquatic insects, and unlike denitrification, previous studies suggest that this pathway may be stimulated by increased availability of P. We tested the hypothesis that the flux of N exported from streams through insect emergence increases relative to flux through denitrification with increasing levels of stream P. We measured emergence and denitrification rates at 7 lowland stream sites in Costa Rica that differ in dissolved P levels and substrate characteristics. Emergence N flux ranged from 2.0 to 16.3% of measured denitrification rates and was not related to any measured physical or chemical variables. Sediment redox conditions were the best predictor of denitrification rates. Our emergence results suggest that most invertebrate biomass production in these streams is consumed by in-stream pr...

Isolation and functional analysis of denitrifiers in an aquifer with high potential for denitrification

Aquifers are among the main freshwater sources. The Raigón aquifer is susceptible to contamination, mainly by nitrate and pesticides, such as atrazine, due to increasing agricultural activities in the area. The capacity of indigenous bacteria to attenuate nitrate contamination in different wells of this aquifer was assessed by measuring denitrification rates with either acetate plus succinate or nitrate amendments. Denitrification activity in nitrate-amended assays was significantly higher than in unamended assays, particularly in groundwater from wells where nitrate concentration was 33.5mgL−1 or lower. Furthermore, groundwater denitrifiers capable of using acetate or succinate as electron donors were isolated, identified by 16S rRNA gene sequencing and evaluated for functional denitrification genes (nirS, nirK and nosZ). Phylogenetic affiliation of 54 isolates showed that all members belonged to nine different genera within the Proteobacteria (Bosea, Ochrobactrum, Azospira, Zoogloea, Acidovorax, Achromobacter, Vogesella, Stenotrophomonas and Pseudomonas). In addition, isolate AR28 that clustered separately from validly described species could potentially belong to a new genus. The majority of the isolates were related to species belonging to previously reported denitrifying genera. However, the phylogeny of the nirS and nosZ genes revealed new sequences of these functional genes. To our knowledge, this is the first isolation and sequencing of the nirS gene from the genus Vogesella, as well as the nosZ gene from the genera Acidovorax and Zoogloea. The results indicated that indigenous bacteria in the Raigón aquifer had the capacity to overcome high nitrate contamination and exhibited functional gene diversity.

Hydrologic connectivity increases denitrification in the hyporheic zone and restored floodplains of an agricultural stream

Stream ecotones, specifically the lateral floodplain and subsurface hyporheic zone, can be important sites for nitrogen (N) removal via denitrification, but their role in streams with constructed floodplains has not been examined. We studied denitrification in the hyporheic zone and floodplains of an agriculturally influenced headwater stream in Indiana, USA, that had floodplains added as part of a “two‐stage ditch” restoration project. To examine the potential for N removal in the hyporheic zone, we seasonally measured denitrification rates and nitrate concentrations by depth into the stream sediments. We found that nitrate concentration and denitrification rates declined with depth into the hyporheic zone, but denitrification was still measureable to a depth of at least 20 cm. We also measured denitrification rates on the restored floodplains over the course of a flood (pre, during, and post‐inundation), and also compared denitrification rates between vegetated and non‐vegetated areas of the floodplain. We found that floodplain denitrification rates increased over the course of a floodplain inundation event, and that the presence of surface water increased denitrification rates when vegetation was present. Stream ecotones in midwestern, agriculturally influenced streams have substantial potential for N removal via denitrification, particularly when they are hydrologically connected with high‐nitrate surface water.

Floodplain restoration enhances denitrification and reach‐scale nitrogen removal in an agricultural stream

Streams of the agricultural Midwest, USA, export large quantities of nitrogen, which impairs downstream water quality, most notably in the Gulf of Mexico. The two-stage ditch is a novel restoration practice, in which floodplains are constructed alongside channelized ditches. During high flows, water flows across the floodplains, increasing benthic surface area and stream water residence time, as well as the potential for nitrogen removal via denitrification. To determine two-stage ditch nitrogen removal efficacy, we measured denitrification rates in the channel and on the floodplains of a two-stage ditch in north-central Indiana for one year before and two years after restoration. We found that instream rates were similar before and after the restoration, and they were influenced by surface water NO3- concentration and sediment organic matter content. Denitrification rates were lower on the constructed floodplains and were predicted by soil exchangeable NO3- concentration. Using storm flow simulations, we found that two-stage ditch restoration contributed significantly to NO3- removal during storm events, but because of the high NO3- loads at our study site, < 10% of the NO3- load was removed under all storm flow scenarios. The highest percentage of NO3- removal occurred at the lowest loads; therefore, the two-stage ditch's effectiveness at reducing downstream N loading will be maximized when the practice is coupled with efforts to reduce N inputs from adjacent fields.

Seasonal and spatial variations of denitrification and denitrifying bacterial community structure in created riverine wetlands

Denitrification rates in wetlands vary spatially and temporally due to a complex set of regulating factors. Also, denitrifying bacterial communities have been suggested to affect their process rates at an ecosystem scale. In this study, we assess the spatio-temporal patterns of denitrifying bacterial communities as well as denitrification rates in two urban created riverine flow-through wetlands. We measured denitrification rates monthly using an acetylene blocking assay along with gene-based denitrifying bacterial community analysis using terminal-restriction fragment length polymorphisms (T-RFLP). The denitrification rates exhibited temporal and spatial differences, ranging from 34 to 660 μg N m −2 h −1. Temporal variation of denitrification primarily depended on temperature and nitrate concentration with high rates in early spring. Significantly lower denitrification rates were seen in the shallow marshes compared to open water inflow and outflow areas and deep marsh areas. Denitrifying bacterial community structure was influenced by vegetation presence, resulting in significant differences between open water and marsh areas. The different spatio-temporal variations between denitrification rates and denitrifying bacterial communities imply that denitrifying bacterial community structure and their denitrification rates are poorly related. Therefore, our results suggest that physiological responses of denitrifiers to environmental conditions rather than a shift of denitrifying bacterial communities regulates the denitrification rate in wetlands.

Online measurement of denitrification rates in aquifer samples by an approach coupling an automated sampling and calibration unit to a membrane inlet mass spectrometry system

Aquifers within agricultural catchments are characterised by high spatial heterogeneity of their denitrification potential. Therefore, simple but sophisticated methods for measuring denitrification rates within the groundwater are crucial for predicting and managing N-fluxes within these anthropogenic ecosystems. Here, a newly developed automated online (15)N-tracer system is presented for measuring (N(2)+N(2)O) production due to denitrification in aquifer samples. The system consists of a self-developed sampler which automatically supplies sample aliquots to a membrane-inlet mass spectrometer. The developed system has been evaluated by a (15)N-nitrate tracer incubation experiment using samples (sulphidic and non-sulphidic) from the aquifer of the Fuhrberger Feld in northern Germany. It is shown that the membrane-inlet mass spectrometry (MIMS) system successfully enabled nearly unattended measurement of (N(2)+N(2)O) production within a range of 10 to 3300 µg N L(-1) over 7 days of incubation. The automated online approach provided results in good agreement with simultaneous measurements obtained with the well-established offline approach using isotope ratio mass spectrometry (IRMS). In addition, three different (15)N-aided mathematical approaches have been evaluated for their suitability to analyse the MIMS raw data under the given experimental conditions. Two approaches, which rely on the measurement of (28)N(2), (29)N(2) and (30)N(2), exhibit the best reliability in the case of a clear (15) N enrichment of evolved denitrification gases. The third approach, which uses only the ratio of (29)N(2)/(28)N(2), overestimates the concentration of labelled denitrification products under these conditions. By contrast, at low (15)N enrichments and low fractions of denitrified gas, the latter approach is on a par with the other two approaches. Finally, it can be concluded that the newly developed system represents a comprehensive and simply applicable tool for the determination of denitrification in aquifers.

Spatial and temporal variation in denitrification and in the denitrifier community in a boreal lake

We investigated the spatial and temporal variation in denitrification rates (isotopepairing technique, IPT) and in the denitrifier community (examination of gene nirK by denaturinggradient gel electrophoresis [DGGE] of microbial DNA) in the sediments of a boreal, clear-water, eutrophic lake using samples collected from shallow littoral, deep littoral and shallow profundal sediments during early summer, mid-summer, autumn and winter. The measured denitrification rates (44 to 613 μmol N m−2 d−1) are among the lowest ever reported from lacustrine sediments. Denitri fi cation rates varied both spatially and temporally, being highest in the profundal zone during midsummer and in the littoral zones during winter. Correlation analyses indicated that these variations were due to variations in the concentrations of nitrate and oxygen in the water overlying the sediment. The structure of the denitrifier community was temporally extremely stable and differed only slightly between the sites. Distance-based linear model (DISTLM) analysis indicated that the observed variation was probably due mainly to variations in the content of organic matter, and in the porosity, of the sediment. The structure of the denitrifier community and the denitrifying activities were uncoupled.

A comparison of different approaches for measuring denitrification rates in a nitrate removing bioreactor

Denitrifying woodchip bioreactors (denitrification beds) are increasingly used to remove excess nitrate ( NO 3 − ) from point-sources such as wastewater effluent or subsurface drains from agricultural fields. NO 3 − removal in these beds is assumed to be due to microbial denitrification but direct measurements of denitrification are lacking. Our objective was to test four different approaches for measuring denitrification rates in a denitrification bed that treated effluent discharged from a glasshouse. We compared these denitrification rates with the rate of NO 3 − removal along the length of the bed. The NO 3 − removal rate was 8.73 ± 1.45 g m −3 d −1. In vitro acetylene inhibition assays resulted in highly variable denitrification rates (DR AI) along the length of the bed and generally 5 times greater than the measured ( NO 3 − —N removal rate. An in situ push–pull test, where enriched N 15 —N O 3 − was injected into 2 locations along the bed, resulted in rates of 23.2 ± 1.43 g N m −3 d −1 and 8.06 ± 1.64 g N m −3 d −1. The denitrification rate calculated from the increase in dissolved N 2 and N 2O concentrations (DR N2) along the length of the denitrification bed was 6.7 ± 1.61 g N m −3 d −1. Lastly, denitrification rates calculated from changes in natural abundance measurements of δ 15N–N 2 and δ 15 N —N O 3 − along the length of the bed yielded a denitrification rate (DR NA) of 6.39 ± 2.07 g m −3 d −1. Based on our experience, DR N 2 measurements were the easiest and most efficient approach for determining the denitrification rate and N 2O production of a denitrification bed. However, the other approaches were useful for testing other hypotheses such as factors limiting denitrification or may be applied to determine denitrification rates in environmental systems different to our study site. DR N 2 does require very careful sampling to avoid atmospheric N 2 contamination but could be used to rapidly determine denitrification rates in a variety of aquatic systems with high N 2 production and even water flows. These measurements demonstrated that the majority of NO 3 − removal was due to heterotrophic denitrification.