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PubMed1994

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 O(2) and NO(3) and rates of nitrification, denitrification, and O(2) consumption as the O(2) concentration in the overlying water was increased from 10 to 600 muM. The model also accurately predicted the response, to increasing O(2) concentrations, of the integrated (micromoles per square meter per hour) rates of nitrification and denitrification. The simulated rates of denitrification of NO(3) diffusing from the overlying water (D(w)) and of NO(3) generated by nitrification within the sediment (D(n)) corresponded to the experimental rates as the O(2) concentration in the overlying water was altered. The predicted D(w) and D(n) rates, as NO(3) 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 NH(4) diffusing from the lower sediment layers and to remove much of the NO(3) produced. In addition to the flux of NH(4) 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 O(2) ( approximately 10 muM), was necessary to reproduce the real data. Spatial separation of the two processes (no toleration by denitrification of O(2)) resulted in too high NO(3) concentrations and too low rates of denitrification.

Journal Article↗

PubMed2005

Maximum rates of nitrate removal in a denitrification wall.

Denitrification walls are constructed by mixing a carbon source such as sawdust into soils through which ground water passes. These systems can reduce nitrate inputs to receiving waters by enhancing denitrification. Maximum rates of nitrate removal by denitrification need to be determined for design purposes. To determine maximum rates of nitrate removal we added excess nitrate (50 mg N L(-1)) to a trench up-gradient of a denitrification wall during a 9-d trial. Bromide (100 g L(-1)) was also added as a conservative tracer. Movement of nitrate and bromide was measured from shallow wells and soil samples were removed for measurements of denitrification, carbon availability, nitrate, and other microbial parameters. Rates of nitrate removal, determined from the ratio of NO3-N to Br and ground water flow, averaged 1.4 g N m(-3) of wall d(-1) and were markedly greater than denitrification rates determined using the acetylene block technique (average: 0.11 g N m(-3) of wall d(-1)). These nitrate removal rates were generally lower than reported in other denitrification walls. Denitrification rates increased when nitrate was added to the laboratory incubations, indicating that despite large nitrate inputs in the field, denitrification remained limited by nitrate. This limitation was partially attributed to nitrate predominantly moving through zones of greater hydraulic conductivity or in the mobile fraction of the ground water and slow diffusion to the immobile fraction where denitrifiers were active.

Adsorption↗

PubMed2006

Effect of denitrification type on pH profiles in the sequencing batch reactor process.

Laboratory batch experiments were conducted to investigate pH profiles during partial and complete denitrification with sufficient organic carbon source. Five stirred tank-type glass vessels, with a 7 L working volume for each, were used as SBR reactors that were all operated in denitrification mode. Five levels of initial proportion of nitrogen substances, i.e. nitrate and nitrite, were used in five reactors, respectively. Results showed that, at given temperature and mixed liquor suspended solids (MLSS), partial denitrification could attain a higher pH value than complete denitrification at the end of denitrification with the same initial NOx- concentration. The larger proportion the nitrite took in initial NOx- concentration, the higher pH peak would be obtained on pH profiles during denitrification despite the same total alkalinity produced. It was found that different types of alkalinity were produced during biological denitrification with different nitrogen substances. Partial denitrification could more carbonate alkalinity produce than complete denitrification. Furthermore, some characteristic points were identified on pH profiles which could indicate the disappearance of not only nitrate, but also nitrite in system. When computers are used to detect these features, they can provide rapid, real-time information, regarding the biological state of the system.

Biomass↗

PubMed1977

Light-activated, -inhibited and -independent denitrification by a denitrifying phototrophic bacterium.

Effects of illumination on denitrification by a freshly isolated denitrifying phototrophic bacterium were investigated. Denitrification activity was induced when cells were grown in either light or darkness in the presence of nitrate without oxygen. Denitrification of nitrate with malate as the electron donor by cells at a phase of exponential growth occurred independently of illumination while that by cells in a stationary phase was activated. Effects of illumination on denitrification varied with electron donors. Using malate or succinate, denitrification by cells in a stationary phase was accelerated by illumination, inhibited when glucose or lactate was used, and independent of illumination when pyruvate was used. Denitrification by cells in an exponential phase was independent of illumination when succinate, malate or pyruvate was used and inhibited by it when glucose or lactate was used. Effects of illumination on the denitrification of nitrite were similar to those involving nitrate. Effects of various inhibitors on denitrification were examined in light-succinate and dark-lactate systems. Differences between the two systems are discussed.

Aerobiosis↗

PubMed2003

Application of polyhydroxyalkanoates for denitrification in water and wastewater treatment.

Application of polyhydroxyalkanoates (PHAs) and related biodegradable polymers has gained momentum in various areas of biotechnology. A promising application that started appearing in the past decade is the use of PHAs as the solid substrate for denitrification of water and wastewater. This type of denitrification, termed here "solid-phase denitrification", has several advantages over the conventional system supplemented with liquid organic substrate. PHAs serve not only as constant sources of reducing power for denitrification but also as solid matrices favorable for development of microbial films. In addition, in contrast to conventional processes, the use of PHAs has no potential risk of release of dissolved organic carbon with the resultant deterioration of effluent water quality. If the production cost of PHAs can be brought down, its application to the denitrification process will become economically more promising. A number of PHA-degrading denitrifying bacteria have been isolated and characterized from activated sludge and continuous flow-bed reactors for denitrification with PHAs. Most of these isolates have been assigned phylogenetically to members of beta-Proteobacteria, especially those of the family Comamonadaceae. The metabolic and regulatory relationships between PHA degradation and denitrification, and the interactive relationship between PHA-degrading cells and the solid surface structure are important subjects awaiting future studies, which would provide a new insight into our comprehensive understanding of the solid-phase denitrification process.

Betaproteobacteria↗

PubMed2003

Addition of trace metals increases denitrification rate in closed marine systems.

We investigated the effect of trace metals (Fe, Mn, Cu, Zn and Mo) on the denitrification unit at the Montreal Biodome. Two dosages of the five trace metals were tested on a denitrifying bacterial population which was extracted from the denitrification unit and cultured in 250 mL chemostats with artificial seawater. The low dosage showed a 20% increase in the denitrification rate whereas the high dosage had a more pronounced effect with a 250% increase. No increase in bacterial growth was observed, suggesting that the trace metals had an effect on the denitrification activity. When the trace metals were tested separately, only iron had a significant effect similar to the increase in the denitrification rate observed when the five trace metals were added. The combination of Fe and Mn caused a small but significant increase compared to the five trace metals. We then tested the effect of adding Fe, Mn and Cu to the denitrification unit at the Montreal Biodome. A high dosage of these trace metals showed a 250% increase in the denitrification rate, which went from 200 to 700 g NO(x)-N/d. Our results showed that the addition of trace metals is crucial for denitrification activities.

Ecosystem↗

PubMed2004

Comparison of different carbon sources for ground water denitrification.

This study presents a comparison of denitrification rates and denitrification stoichiometry when using different sources of carbon. Denitrification tests were carried out in test bottles containing water and soil samples acquired at a nitrate-contaminated site and supplemented with either sucrose, acetate or ethanol. The tests demonstrated nitrate removal in all carbon source supplemented bottles. The rate of denitrification and the required amount of a carbon source, however, depended on the choice of substrate. Ethanol and acetate were found to provide the highest denitrification rate, that of 1.4 - 1.6 mg-N l(-1) d(-1). Sucrose-supplemented bottles demonstrated a significantly slower denitrification rate, that of 0.6 - 0.9 mg-N l(-1) d(-1). In addition to slow denitrification rates, sucrose-supported denitrification required more carbon source.

Acetates↗

PubMed2001

Theoretical stoichiometry of biological denitrifications.

Biological denitrification is a phenomenon widely present in soil, water, and wastewater treatment ecosystems. Based on McCarty's half-reaction theory, taking the fraction (f(s)) of electron donor coupled to cells (CalphaHbetaOepsilonNdelta) synthesis as the basic parameter, we developed new stoichiometric equations for biological denitrifications. These denitrifications are complete denitrifications of nitrate with, respectively, nitrate and ammonia as the nitrogen source, complete denitrification of nitrite and reduction of nitrate to nitrite with various kinds of organic carbon sources. Taking the concept that all electron donors in stoichiometric equations are equivalent to 1/4 moles of COD, we discussed the stoichiometric equations and relationships of biological denitrifications. Some COD-independent stoichiometric relationships and parameters for biological denitrifications are discovered. The generalized stoichiometric equations and stoichiometric relationships are discussed in detail by citing the example of CalphaHbetaOepsilonNdelta = C5H7O2N . Some empirical values of f(s) from previous reports are calculated by using the theoretical stoichiometric coefficients and relationships. The effects of COD/N-NO3 on the denitrification behavior and the cell yield coefficient Yx/c are also discussed here. Based on this method, the parameter of f(s) and the balance of electron flow besides the mass balance appear very important for the stoichiometry of bioprocesses. The new method for the stoichiometric analysis of bioprocesses may be termed electronic stoichiometry.

Chemical Phenomena↗

PubMed2006

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.

Alteromonadaceae↗

PubMed1982

Denitrification in marine sediment: measurement of capacity and estimate of in situ rate.

The acetylene block technique was used to assay denitrification in undisturbed sediment cores of an intertidal mud flat. Nitrogen loss due to this process was estimated at 1 to 41 kg of N/hectare (ha) per year. Anaerobic nitrate-saturated slurry of the same sediment had a denitrification capacity of 1,026 kg of N/ha per year. The acetylene block technique failed at low nitrate concentrations, so that denitrification at average in situ nitrate concentrations could not be determined. Denitrification followed zero-order kinetics at nitrate concentrations high enough to allow successful blockage of N(2)O reduction. Thus, an estimate of in situ rates based on kinetic parameters and in situ nitrate concentrations was impossible. No denitrification was observed in a slurry of the top 1.5 cm under aerated conditions and nitrate saturation. In undisturbed sediment, significant denitrification occurred in few discrete sites within a matrix of undetectable or low activity. Despite numerous errors contributing to the uncertainty of the estimate of in situ rates, the result obtained by this method was considered more valuable than the determination of denitrification capacities. Methods which include severe changes of physical and chemical parameters may frequently lead to overestimates of denitrification rates.

Journal Article↗

PubMed1986

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

The source of N(2)O 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 C(2)H(2) inhibition is proposed for distinguishing among gaseous N losses from soils. Denitrification N(2)O was estimated from 24-h laboratory incubations in which nitrification was inhibited by 10-Pa C(2)H(2). Nitrification N(2)O was estimated from the difference between N(2)O production under no C(2)H(2) and that determined for denitrification. Denitrification N(2) was estimated from the difference between N(2)O production under 10-kPa C(2)H(2) and that under 10 Pa. Laboratory estimates of N(2)O production were significantly correlated with in situ N(2)O 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 NO(3) was high. In contrast, denitrification N(2)O was most important on poorly drained sites near the stream of the same watershed. Distinction between N(2)O production from nitrification and denitrification was corroborated by correlations between denitrification N(2)O and water-filled pore space and between nitrification N(2)O and ambient NO(3). This technique permits qualitative study of environmental parameters that regulate gaseous N losses via denitrification and nitrification.

Journal Article↗

PubMed1996

Dynamics of denitrification activity of Paracoccus denitrificans in continuous culture during aerobic-anaerobic changes.

Induction and repression of denitrification activity were studied in a continuous culture of Paracoccus denitrificans during changes from aerobic to anaerobic growth conditions and vice versa. The denitrification activity of the cells was monitored by measuring the formation of denitrification products (nitrite, nitric oxide, nitrous oxide, and dinitrogen), individual mRNA levels for the nitrate, nitrite, and nitrous oxide reductases, and the concentration of the nitrite reductase enzyme with polyclonal antibodies against the cd1-type nitrite reductase. On a change from aerobic to anaerobic respiration, the culture entered an unstable transition phase during which the denitrification pathway became induced. The onset of this phase was formed by a 15- to 45-fold increase of the mRNA levels for the individual denitrification enzymes. All mRNAs accumulated during a short period, after which their overall concentration declined to reach a stable value slightly higher than that observed under aerobic steady-state conditions. Interestingly, the first mRNAs to be formed were those for nitrate and nitrous oxide reductase. The nitrite reductase mRNA appeared significantly later, suggesting different modes of regulation for the three genes. Unlike the mRNA levels, the level of the nitrite reductase protein increased slowly during the anaerobic period, reaching a stable value about 30 h after the switch. All denitrification intermediates could be observed transiently, but when the new anaerobic steady state was reached, dinitrogen was the main product. When the anaerobic cultures were switched back to aerobic respiration, denitrification of the cells stopped at once, although sufficient nitrite reductase was still present. We could observe that the mRNA levels for the individual denitrification enzymes decreased slightly to their aerobic, uninduced levels. The nitrite reductase protein was not actively degraded during the aerobic period.

Aerobiosis↗

PubMed1997

Cell biology and molecular basis of denitrification.

Denitrification is a distinct means of energy conservation, making use of N oxides as terminal electron acceptors for cellular bioenergetics under anaerobic, microaerophilic, and occasionally aerobic conditions. The process is an essential branch of the global N cycle, reversing dinitrogen fixation, and is associated with chemolithotrophic, phototrophic, diazotrophic, or organotrophic metabolism but generally not with obligately anaerobic life. Discovered more than a century ago and believed to be exclusively a bacterial trait, denitrification has now been found in halophilic and hyperthermophilic archaea and in the mitochondria of fungi, raising evolutionarily intriguing vistas. Important advances in the biochemical characterization of denitrification and the underlying genetics have been achieved with Pseudomonas stutzeri, Pseudomonas aeruginosa, Paracoccus denitrificans, Ralstonia eutropha, and Rhodobacter sphaeroides. Pseudomonads represent one of the largest assemblies of the denitrifying bacteria within a single genus, favoring their use as model organisms. Around 50 genes are required within a single bacterium to encode the core structures of the denitrification apparatus. Much of the denitrification process of gram-negative bacteria has been found confined to the periplasm, whereas the topology and enzymology of the gram-positive bacteria are less well established. The activation and enzymatic transformation of N oxides is based on the redox chemistry of Fe, Cu, and Mo. Biochemical breakthroughs have included the X-ray structures of the two types of respiratory nitrite reductases and the isolation of the novel enzymes nitric oxide reductase and nitrous oxide reductase, as well as their structural characterization by indirect spectroscopic means. This revealed unexpected relationships among denitrification enzymes and respiratory oxygen reductases. Denitrification is intimately related to fundamental cellular processes that include primary and secondary transport, protein translocation, cytochrome c biogenesis, anaerobic gene regulation, metalloprotein assembly, and the biosynthesis of the cofactors molybdopterin and heme D1. An important class of regulators for the anaerobic expression of the denitrification apparatus are transcription factors of the greater FNR family. Nitrate and nitric oxide, in addition to being respiratory substrates, have been identified as signaling molecules for the induction of distinct N oxide-metabolizing enzymes.

Amino Acid Sequence↗

PubMed2006

Modeling denitrification in terrestrial and aquatic ecosystems at regional scales.

Quantifying where, when, and how much denitrification occurs on the basis of measurements alone remains particularly vexing at virtually all spatial scales. As a result, models have become essential tools for integrating current understanding of the processes that control denitrification with measurements of rate-controlling properties so that the permanent losses of N within landscapes can be quantified at watershed and regional scales. In this paper, we describe commonly used approaches for modeling denitrification and N cycling processes in terrestrial and aquatic ecosystems based on selected examples from the literature. We highlight future needs for developing complementary measurements and models of denitrification. Most of the approaches described here do not explicitly simulate microbial dynamics, but make predictions by representing the environmental conditions where denitrification is expected to occur, based on conceptualizations of the N cycle and empirical data from field and laboratory investigations of the dominant process controls. Models of denitrification in terrestrial ecosystems include generally similar rate-controlling variables, but vary in their complexity of the descriptions of natural and human-related properties of the landscape, reflecting a range of scientific and management perspectives. Models of denitrification in aquatic ecosystems range in complexity from highly detailed mechanistic simulations of the N cycle to simpler source-transport models of aggregate N removal processes estimated with empirical functions, though all estimate aquatic N removal using first-order reaction rate or mass-transfer rate expressions. Both the terrestrial and aquatic modeling approaches considered here generally indicate that denitrification is an important and highly substantial component of the N cycle over large spatial scales. However, the uncertainties of model predictions are large. Future progress will be linked to advances in field measurements, spatial databases, and model structures.

Agriculture↗

PubMedDate not supplied

Seasonal dynamics of denitrification along topohydrosequences in three different riparian wetlands.

We investigated the seasonal patterns of denitrification rates and potentials in soil profiles along the topohydrosequence formed at the upland-wetland interface in three riparian wetlands with different vegetation cover (i.e., forest, understory vegetation, and grass). Denitrification was measured using the acetylene inhibition method on soil cores and slurries, which provided a means of comparing the relative activity of this process in different locations. We evaluated, on a seasonal basis, the respective importance of the vegetative cover and the hydromorphic gradient as factors limiting denitrification. Regardless of the season, vegetation type, or lateral position along each topohydrosequence in the riparian wetlands, strong significant gradients of both in situ and potential denitrification rates were measured within a soil profile. Results confirm that the upper organic soil horizon is the most active, when in contact with the ground water. In deeper soil horizons, denitrification activity was low (from 0.004 to 0.5 mg N kg(-1) dry soil d(-1)), but contributed significantly to the reduction of ground water NO3- load along the riparian ground water flowpath (from 9.32 to 0.98 mg NO3-N L(-1)). Along the soil topohydrosequence, the denitrifying community of the upper soil horizons did not vary significantly on a seasonal basis despite the large seasonal ground water fluctuations. Along each topohydrosequence, the denitrification-limiting factor gradually shifted from anaerobiosis to NO3- supply. In situ denitrification rates in the forested, understory vegetation and grass sites were not significantly different. This result emphasizes the importance of the topography of the valley rather than the vegetation cover in controlling denitrification activity in riparian wetlands.

France↗

PubMedDate not supplied

Assessment of denitrification gaseous end-products in the soil profile under two water table management practices using repeated measures analysis.

The denitrification process and nitrous oxide (N2O) production in the soil profile are poorly documented because most research into denitrification has concentrated on the upper soil layer (0-0.15 m). This study, undertaken during the 1999 and 2000 growing seasons, was designed to examine the effects of water table management (WTM), nitrogen (N) application rate, and depth (0.15, 0.30, and 0.45 m) on soil denitrification end-products (N2O and N2) from a corn (Zea mays L.) field. Water table management treatments were free drainage (FD) with open drains and subirrigation (SI) with a target water table depth of 0.6 m. Fertility treatments (ammonium nitrate) were 120 kg N ha(-1) (N120) and 200 kg N ha(-1) (N200). During both growing seasons greater denitrification rates were measured in SI than in FD, particularly in the surface soil (0-0.15 m) and at the intermediate (0.15-0.30 m) soil depths under N200 treatment. Greater denitrification rates under the SI treatment, however, were not accompanied with greater N2O production. The decrease in N2O production under SI was probably caused by a more complete reduction of N2O to N2, which resulted in lower N2O to (N2O + N2) ratios. Denitrification rate, N2O production and N2O to (N2O + N2) ratios were only minimally affected by N treatments, irrespective of sampling date and soil depth. Overall, half of the denitrification occurred at the 0.15- to 0.30- and 0.30- to 0.45-m soil layers, and under SI, regardless of fertility treatment level. Consequently, sampling of the 0- to 0.15-m soil layer alone may not give an accurate estimation of denitrification losses under SI practice.

Agriculture↗

PubMedDate not supplied

In situ ground water denitrification in stratified, permeable soils underlying riparian wetlands.

The ground water denitrification capacity of riparian zones in deep soils, where substantial ground water can flow through low-gradient stratified sediments, may affect watershed nitrogen export. We hypothesized that the vertical pattern of ground water denitrification in riparian hydric soils varies with geomorphic setting and follows expected subsurface carbon distribution (i.e., abrupt decline with depth in glacial outwash vs. negligible decline with depth in alluvium). We measured in situ ground water denitrification rates at three depths (65, 150, and 300 cm) within hydric soils at four riparian sites (two per setting) using a 15N-enriched nitrate "push-pull" method. No significant difference was found in the pattern and magnitude of denitrification when grouping sites by setting. At three sites there was no significant difference in denitrification among depths. Correlations of site characteristics with denitrification varied with depth. At 65 cm, ground water denitrification correlated with variables associated with the surface ecosystem (temperature, dissolved organic carbon). At deeper depths, rates were significantly higher closer to the stream where the subsoil often contains organically enriched deposits that indicate fluvial geomorphic processes. Mean rates ranged from 30 to 120 microg N kg(-1) d(-1) within 10 m versus <1 to 40 microg N kg(-1) d(-1) at >30 m from the stream. High denitrification rates observed in hydric soils, down to 3 m within 10 m of the stream in both alluvial and glacial outwash settings, argue for the importance of both settings in evaluating the significance of riparian wetlands in catchment-scale N dynamics.

Ecosystem↗

PubMed2001

Simultaneous nitrification and denitrification in biofilters with real time aeration control.

The aim of this article is to present a new biological aerated filter (BAF) for nitrogen removal based on simultaneous nitrification and denitrification. Contrary to the systems which integrate both an aerated and a non-aerated zone to allow complete nitrogen removal in one compact or two different units (pre-denitrification and nitrification), this upflow BAF system is based on the principle of simultaneous nitrification and denitrification since the filter is completely aerated. The denitrification process is possible due to the diffusion effect which dominates biofilm processes. The real time aeration control allows us to maintain a low dissolved oxygen value (0.5 to 3 mg O2/l). In this case, the biofilm will not be fully (or less) penetrated with oxygen and denitrification will be carried out in a large part of the biofilm. Therefore, nitrification and denitrification is running simultaneously in different depths of the biofilm. By using 50% less air this BAF gave the same results (less than 20 mg TN/l) on pilot plant as a classical nitrification and denitrification BAF (Toettrup et al., 1994). Less recirculation was necessary to achieve the same denitrification.

Biofilms↗