Nitrogen mass balances and denitrification rates in central Ontario Lakes

Nitrogen mass balances for seven unproductive lakes and 20 forested catchments in central Ontario were measured between 1977 and 1989. Average annual lake denitrification rates calculated with the N/P ratio method were strongly correlated with summer anoxic factor (extent of surficial sediment anoxia) whereas denitrification rates calculated with a²¹⁰Pb sediment N accumulation method were poorly correlated with the anoxic factor suggesting that the N/P method is superior. Substantial denitrification occurred in all lakes — an average of 36% of TN inputs or 75% of the net gain. On a regional area-weighted basis, 67% of bulk atmospheric TN deposition was stored or denitrified terrestrially, 12% was denitrified in lakes, 4% was stored in lake sediments, and 17% was exported from lakes. N/P ratios were generally less in streams than in precipitation suggesting preferential N retention in catchments, whereas the N/P ratios in lake outputs were slightly higher than lake input ratios, suggesting preferential P retention in lakes. This is consistent with the notion that P-limited lakes can exist adjacent to N-limited forests.     

Tree species, root decomposition and subsurface denitrification potential in riparian wetlands

Patches of organic matter have been found to be important `hotspots' of denitrification in both surface and subsurface soils, but the factors controlling the formation and maintenance of these patches are not well established. We compared the concentration of patches of organic matter and root biomass in the subsurface (saturated zone) beneath poorly drained riparian wetland soils at four sites in Rhode Island, USA - two dominated by red maple (Acer rubrum) and two dominated by white pine (Pinus strobus). Denitrification enzyme activity (DEA) and carbon (C) content of patch material were compared between sites and between patches with different visual characteristics. Root decomposition was measured in an 8-week ex-situ incubation experiment that compared the effects of water content, root species, and soil matrix origin on CO₂ evolution. We observed significantly greater concentrations of patches at 55 cm at one red maple site than all other sites. DEA and percent C in patches was generally higher in patches than matrix soil and did not vary between sites or by patch type. White pine roots decomposed at a faster rate than red maple roots under unsaturated conditions. Our results suggest that faster root decomposition could result in lower concentrations of patches of organic material in subsurface soils at sites dominated by white pine. Tree species composition and root decomposition may play a significant role in the formation of patches and the creation and maintenance of groundwater denitrification hotspots in the subsurface of riparian wetlands. Abbreviations: DEA – denitrification enzyme activity; DOC – dissolved organic carbon; PD – poorly drained; RM-1 – red maple-1 site; RM-2 – red maple-2 site; WP-1 – white pine-1 site; WP-2 – white pine-2 site.     

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

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

The potential for nitrification and nitrate uptake in the rhizosphere of wetland plants: a modelling study

BACKGROUND AND AIMS: It has recently found that lowland rice grown hydroponically is exceptionally efficient in absorbing NO3-, raising the possibility that rice and other wetland plants growing in flooded soil may absorb significant amounts of NO3- formed by nitrification of NH4+ in the rhizosphere. This is important because (a) this NO3- is otherwise lost through denitrification in the soil bulk; and (b) plant growth and yield are generally improved when plants absorb their nitrogen as a mixture of NO3- and NH4+ compared with growth on either N source on its own. A mathematical model is developed here with which to assess the extent of NO3- absorption from the rhizosphere by wetland plants growing in flooded soil, considering the important plant and soil processes operating. METHODS: The model considers rates of O2 transport away from an individual root and simultaneous O2 consumption in microbial and non-microbial processes; transport of NH4+ towards the root and its consumption in nitrification and uptake at the root surface; and transport of NO3- formed from NH4+ towards the root and its consumption in denitrification and uptake by the root. The sensitivity of the model's predictions to its input parameters is tested over the range of conditions in which wetland plants grow. KEY RESULTS: The model calculations show that substantial quantities of NO3- can be produced in the rhizosphere of wetland plants through nitrification and taken up by the roots under field conditions. The rates of NO3- uptake can be comparable with those of NH4+. The model also shows that rates of denitrification and subsequent loss of N from the soil remain small even where NO3- production and uptake are considerable. CONCLUSIONS: Nitrate uptake by wetland plants may be far more important than thought hitherto. This has implications for managing wetland soils and water, as discussed in this paper.     

Removal of highly elevated nitrate from drinking water by pH-heterogenized heterotrophic denitrification facilitated with ferrous sulfide-based autotrophic denitrification

The performance of acetic acid-supported pH-heterogenized heterotrophic denitrification (HD) facilitated with ferrous sulfide-based autotrophic denitrification (AD) was investigated in upflow activated carbon-packed column reactors for reliable removal of highly elevated nitrate (42 mg NO₃-N l⁻¹) in drinking water. The use of acetic acid as substrate provided sufficient internal carbon dioxide to completely eliminate the need of external pH adjustment for HD, but simultaneously created vertically heterogenized pH varying from 4.8 to 7.8 in the HD reactor. After 5-week acclimation, the HD reactor developed a moderate nitrate removal capacity with about one third of nitrate removal occurring in the acidic zone (pH 4.8–6.2). To increase the treatment reliability, acetic acid-supported HD was operated under 10% carbon limitation to remove >85% of nitrate, and ferrous sulfide-based AD was supplementally operated to remove residual nitrate and formed nitrite without excess of soluble organic carbon, nitrite or sulfate in the final effluent.     

Simultaneous occurrence of denitrification and nitrate ammonification in sediments of the French Mediterranean Coast

Dissimilatory nitrate reductions in coastal marine sediment of Carteau Cove (French Mediterranean Coast) were studied between April 1993 and July 1994. Simultaneous determination of denitrification and dissimilatory nitrate reduction to ammonium was achieved by using a combination of acetylene blockage and 15N techniques. After short incubations (maximum 5 h), a part of 15N labelled nitrate added to the sediment was recovered as ammonium without incorporation in organic matter. The result indicate that a fraction of nitrate was reduced to ammonium by a dissimilatory mechanism instead of denitrifying. Denitrifying and nitrate ammonifying activities ranged from 0 to 19.8 μmol l-1 d-1 and from 2.3 to 83.2 μmol l-1 d-1, respectively. Denitrification rates were highest in early spring whereas nitrate ammonification were highest in fall. The recovery of nitrate reduced as N2O-N plus ammonium was between 40 and 100%, the highest nitrogen losses were recorded in July. Depending on the station and time of year denitrification accounted for between 0 and 43% of the total nitrate reduction whereas dissimilatory nitrate reduction to ammonium (DNRA) accounted for between 18 and 100%. The reduction rate data suggest that the pathway of nitrate reduction to ammonium may be important in coastal sediments. This revised version was published online in July 2006 with corrections to the Cover Date.     

Nitrification, denitrification, and nitrate ammonification in sediments of two coastal lagoons in Southern France

Summary Seasonal and diurnal variations in sediment-water fluxes of O₂, NO ₃ ^– , and NH ₄ ⁺ as well as rates of nitrification, denitrification, and nitrate ammonification were determined in two different coastal lagoons of southern France: The seagrass (Zostera noltii) dominated tidal Bassin d'Arcachon and the dystrophic Etang du Prévost. Overall, denitrification rates in both Bassin d'Arcachon (<0.4 mmol m^–2 d^–1) and Etang du Prévost (<1 mmol m^–2 d^–1) were low. This was mainly caused by a combination of low NO ₃ ^– concentrations in the water column and a low nitrification activity within the sediment. In both Bassin d'Arcachon and Etang du Prévost, rates of nitrate ammonification were quantitatively as important as denitrification.Denitrification played a minor role as a nitrogen sink in both systems. In the tidal influenced Bassin d'Arcachon, Z. noltii was quantitatively more important than denitrification as a nitrogen sink due to the high assimilation rates of the plants. Throughout the year, Z. noltii stabilized the mudflats of the bay by its well- developed root matrix and controlled the nitrogen cycle due to its high uptake rates. In contrast, the lack of rooted macrophytes, and dominance of floating macroalgae, made nitrogen cycling in Etang du Prévost more unstable and unpredictable. Inhibition of nitrification and denitrification during the dystrophic crisis in the summer time increased the inorganic nitrogen flux from the sediment to the water column and thus increased the degree of benthic-pelagic coupling within this bay. During winter, however, benthic microalgae colonizing the sediment surface changed the sediment in the lagoon from being a nitrogen source to the over lying water to being a sink due to their high assimilation rates. It is likely, however, that this assimilated nitrogen is liberated to the water column at the onset of summer thereby fueling the extensive growth of the floating macroalgae, Ulva sp. The combination of a high nitrogen coupling between sediment and water column, little water exchange and low denitrification rates resulted in an unstable system with fast growing algal species such as phytoplankton and floating algae.     

Paracoccus denitrificans for the effluent recycling during continuous denitrification of liquid food

Nitrate is an undesirable component of several foods. A typical case of contamination with high nitrate contents is whey concentrate, containing nitrate in concentrations up to 25 l. The microbiological removal of nitrate by Paracoccus denitrificans under formation of harmless nitrogen in combination with a cell retention reactor is described here. Focus lies on the resource‐conserving design of a microbal denitrification process. Two methods are compared. The application of polyvinyl alcohol‐immobilized cells, which can be applied several times in whey feed, is compared with the implementation of a two step denitrification system. First, the whey concentrate's nitrate is removed by ion exchange and subsequently the eluent regenerated by microorganisms under their retention by crossflow filtration. Nitrite and nitrate concentrations were determined by reflectometric color measurement with a commercially available Reflectoquant® device. Correction factors for these media had to be determined. During the pilot development, bioreactors from 4 to 250 mg·L^?1 and crossflow units with membrane areas from 0.02 to 0.80 m² were examined. Based on the results of the pilot plants, a scaling for the exemplary process of denitrifying 1,000 tons per day is discussed. © 2010 American Institute of Chemical Engineers Biotechnol. Prog. 2010     

Assessment of denitrification in organogenic forest soil by regulating factors

One of the dominating sources of the increase in atmospheric N₂O are elevated emissions from terrestrial ecosystems. The N₂O production in soil is probably dominated by two microbial processes, i.e. nitrification and denitrification, and their N₂O production is regulated by an array of soil and environmental factors. This contributes to the large both spatial and temporal variability of emission rates measured in the field. To obtain reasonable estimates of large-scale emission rates this variability has to be dealt with, and one possible way could be to relate measured emission rates to relevant soil and/or environmental factors. In this study the relationship between soil factors (temperature, soil organic matter, soil moisture, pH, and NH₄ ⁺, NO₃ ^- and NO₂ ^- contents) and the denitrification activity in organogenic forest soil was investigated. About 40% or less of the variation of the denitrification rates could be accounted for by soil factors, either by multiple regression or by partial least squares analysis of latent variables (PLS analysis). Interpretation of the relationships obtained was, however, difficult since many of them were unexpected, and some of the soil factors were internally related. In conclusion, the possibility to obtain significant and useful relationships between momentary denitrification rates and soil factors for spatial and temporal extrapolations was limited in this system.     

Effect of aeration on denitrification byOchrobactrum anthropi SY509

Aeration was found to affect the biological denitrification byOchrobactrum anthropi SY509. Although cell growth was vigorous under 1 vvm of aeration and an agitation speed of 400 rpm in a 3-L jar fermentor, almost no nitrate was removed. Yet under low agitation speeds (100, 200, and 300 rpm), denitrification occurred when the dissolved oxygen was exhausted shortly after the inoculation of the microorganism.Ochrobactrum anthropi SY509 was found to express highly active denitrifying enzymes under anaerobic conditions. The microorganism also synthesized denitrifying enzymes under aerobic conditions (1 vvm and 400 rpm), yet their activity was only 60% of the maximum level under anaerobic conditions and the nitrate removal efficiency was merely 15%. However, although the activities of the denitrifying enzymes were inhibited in the presence of oxygen, they were fully recovered when the conditions were switched to anaerobic conditions.     

Nitrification and denitrification by Thiosphaera pantotropha in aerobic chemostat cultures

Abstract: Thiosphaera pantotropha has been reported to denitrify aerobically and nitrify heterotrophically. However, recent evidence has indicated that these properties (particularly aerobic denitrification) have been lost. The occurrence and levels of aerobic denitrification and heterotrophic nitrification by T. pantotropha in chemostat cultures have therefore been re-evaluated. Only low nitrate reduction rates were observed: the apparent nitrogen loss was of the same order of magnitude as the combined error in the calculated nitrogen consumption. However, ¹⁵N mass spectrometry revealed low aerobic denitrification rates (about 10% of the rates originally published by this group). Heterotrophic nitrification rates were about a third of previous observations. N₂ and N₂O were both produced from NH₄⁺, NO₃⁻ and NO₂⁻. Periplasmic nitrate reductase was present in aerobically grown cells.     

Benthic organic carbon influences denitrification in streams with high nitrate concentration

1. Anthropogenic activities have increased reactive nitrogen availability, and now many streams carry large nitrate loads to coastal ecosystems. Denitrification is potentially an important nitrogen sink, but few studies have investigated the influence of benthic organic carbon on denitrification in nitrate‐rich streams. 2. Using the acetylene‐block assay, we measured denitrification rates associated with benthic substrata having different proportions of organic matter in agricultural streams in two states in the mid‐west of the U.S.A., Illinois and Michigan. 3. In Illinois, benthic organic matter varied little between seasons (5.9–7.0% of stream sediment), but nitrate concentrations were high in summer (>10 mg N L⁻¹) and low (<0.5 mg N L⁻¹) in autumn. Across all seasons and streams, the rate of denitrification ranged from 0.01 to 4.77 μg N g⁻¹ DM h⁻¹ and was positively related to stream‐water nitrate concentration. Within each stream, denitrification was positively related to benthic organic matter only when nitrate concentration exceeded published half‐saturation constants. 4. In Michigan, streams had high nitrate concentrations and diverse benthic substrata which varied from 0.7 to 72.7% organic matter. Denitrification rate ranged from 0.12 to 11.06 μg N g⁻¹ DM h⁻¹ and was positively related to the proportion of organic matter in each substratum. 5. Taken together, these results indicate that benthic organic carbon may play an important role in stream nitrogen cycling by stimulating denitrification when nitrate concentrations are high.     

Subsurface denitrification in a forest riparianzone: Interactions between hydrology and supplies ofnitrate and organic carbon

The influence of hydrology andpatterns of supply of electron donors and acceptors onsubsurface denitrification was studied in a forestriparian zone along the Boyne River in southernOntario that received high nitrogen inputs from a sandaquifer. Two hypotheses were tested: (1) subsurfacedenitrification is restricted to localized zones ofhigh activity; (2) denitrification zones occur atsites where groundwater flow paths transportNO₃ ^– to supplies of available organiccarbon. A plume of nitrate-rich groundwater withconcentrations of 10–30 mg N L^–1 flowed laterallyat depths of 1.5–5 m in sands beneath peat for ahorizontal distance of 100–140 m across the riparianzone to within 30–50 m of the river. In situ acetyleneinjections to piezometers revealed that significantdenitrification was restricted to a narrow zone ofsteep NO₃ ^– and N₂O decline at theplume margins. The location of these denitrificationsites in areas with steep gradients of groundwater DOCincrease supported hypothesis 2. Many of thesedenitrification hotspots occurred near interfacesbetween sands and either peats or buried river channeldeposits. Field experiments involving in situadditions of either glucose or NO₃ ^– topiezometers indicated that denitrification wasC-limited in a large subsurface area of the riparianzone, and became N-limited beyond the narrow zone ofNO₃ ^– consumption. These data suggest thatdenitrification may not effectively removeNO₃ ^– from groundwater transported at depththrough permeable riparian sediments unlessinteraction occurs with localized supplies of organicmatter.     

Whole-system estimation of denitrification in a plains river: a comparison of two methods

Whole-system denitrification in the South Platte River was measured over a 13-month period using an open-channel N₂ method and mass-balance measurements. Concentrations of dissolved N₂ were measured with high precision by membrane-inlet mass spectrometry and estimates of denitrification were based on the mass flux of N₂, after correction for reaeration and groundwater flux. Open-channel estimates of denitrification ranged from 0 to 3.08 g N m^–2 d^–1 and the mean annual rate was 1.62 g N m^–2 d^–1, which corresponds to removal of approximately 34% of the nitrate transported by the river over a distance of 18.5 km. Over the same period of time, estimates of denitrification based on mass-balance measurements ranged from 0.29 to 5.25 g N m^–2 d^–1 and the mean annual rate was 2.11 g N m^–2 d^–1. The two methods revealed similar seasonal patterns of denitrification the highest rates were measured from late April to August and the lowest rates were in winter. Both methods provide whole-system estimates of denitrification in running waters; where reaeration rate coefficients are low and flux of groundwater is well quantified, the open-channel method has fewer sources of uncertainty and is easier to implement.     

Characteristics of bacteria showing high denitrification activity in saline wastewater

AIMS: Denitrification efficiency at 10% salinity was compared with that at 2% salinity. The characteristics of bacterial strains isolated from the denitrification system, where an improvement of denitrification efficiency was observed at a high salinity were investigated. METHODS AND RESULTS: Two continuous feeding denitrification systems for saline solutions of 2% and 10% salinity, were operated. Denitrification efficiency at 10% salinity was higher than that at 2% salinity. The bacterial strains were isolated using the trypticase soy agar (TSA) medium at 30 degrees C. The phylogenetic analysis of 16S rRNA gene sequences of isolates indicated that halophilic species were predominant at 10% salinity. CONCLUSIONS: The improvement of denitrification efficiency at a high salinity was demonstrated. The strains isolated from the denitrifying system with 10% salinity were halophilic bacteria, Halomonas sp. and Marinobacter sp., suggesting that these bacteria show a high denitrifying activity at 10% salinity. SIGNIFICANCE AND IMPACT OF THE STUDY: The long-term acclimated sludge used in this study resulted in high denitrification performance at a high salinity, indicating that the design of a high-performance denitrification system for saline wastewater will be possible.     

Denitrification in a South Louisiana wetland forest receiving treated sewage effluent

Although denitrification has the potential to reduce nitrate (NO ₃ ^– ) pollution of surface waters, the quantification of denitrification rates is complex because it requires differentiation from other mechanisms and is highly variable in both space and time. This study first measured potential denitrification rates at a wetland forest site in south Louisiana before receipt of secondary wastewater effluent, and then, following 30 months of effluent application, landscape gradients of dissolved nitrate (NO ₃ ^– ) and nitrous oxide (N₂O) were measured. A computer model was developed to quantify N transformations. Floodwater NO ₃ ^– and N₂O concentrations were higher in the forest receiving effluent than in the adjacent control forest. Denitrification rates of NO ₃ ^– -amended soil cores ranged from 0.03–0.45 g N m^–2 d^–1 with an overall mean of 0.10 g N m^–2 d^–1. Effluent N is being applied at a rate of approximately 0.034 g N m^–2 d^–1, with approximately 95% disappearing along a 1 km transect. In the treatment forest, floodwater NO ₃ ^– concentrations decreased from 1000 M at the inflow point to 50 M along the 1 km transect. Nitrous oxide concentrations increased from 0.25 M to 1.2 M within the first 100 m, but decreased to 0.1 M over the next 900 m. The initial increase in N₂O was presumably a result ofin situ denitrification. Model analyses indicated that denitrification was directly associated with nitrification and was limited by the availability of NO ₃ ^– produced by nitrification. Due to different redox potential optima, coupling of nitrification and denitrification was a function of a balance of environmental conditions that was moderately favorable to both processes. N removal efficiency was largely dependent on the proportion of effluent NH ₄ ⁺ to NO ₃ ^– . When NH ₄ ⁺ /NO ₃ ^– was 1, average N removal efficiency ranged from 95–100%, but ratios that were >1 reduced average efficiencies to as low as 57%. Actual effluent NH ₄ ⁺ /NO ₃ ^– loading ratios at this site are approximately 0.2 and are consistently <1.