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Seasonal Denitrification in Flooded and Exposed Sediments from the Amazon Floodplain at Lago Camaleao

Denitrification processes were measured by the acetylene-blockage technique under changing flood conditions along the aquatic/terrestrial transition zone on the Amazon floodplain at Lago Camaleao, near Manaus, Brazil. In flooded sediments, denitrification was recorded after the amendment with NO3- (100 μmol liter-1) throughout the whole study period from August 1992 to February 1993. It ranged from 192.3 to 640.7 μmol N m-2 h-1 in the 0- to 5-cm sediment layer. Without substrate amendment, denitrification was detected only during low water in November and December 1992, when it occurred at a rate of up to 12.2 μmol N m-2 h-1. Higher rates of denitrification at an average rate of 73.3 μmol N m-2 h-1 were measured in sediments from the shallow lake basin that were exposed to air at low water. N2O evolution was never detected in flooded sediments, but in exposed sediments, it was detected at an average rate of 28.3 μmol N m-2 h-1 during the low-water period. The results indicate that under natural conditions there is denitrification and hence a loss in nitrogen from the Amazon floodplain to the atmosphere. Rates of denitrification in flooded sediments were one to two orders of magnitude smaller than in temperate regions. However, the nitrogen removal of exposed sediments exceeded that of undisturbed wetland soils of temperate regions, indicating a considerable impact of the flood pulse on the gaseous turnover of nitrogen in the Amazon floodplain.

Journal Article↗

Changes in potential denitrification and respiration during the cold storage of soils.

The denitrification potential in moderately fertilized soil sampled four times during 1995 decreased significantly after cold storage, at 4 +/- 2 degrees C for 1 week. Prolonged storage (up to 24 weeks) resulted in a further decrease of denitrification potential which dropped to 38-54% of the original values. Similarly, denitrification potential decreased substantially during the first week of storage in differently fertilized soils. After 24 weeks of storage, denitrification potential dropped to 29-55% of that in fresh soils. The effects of storage at 4 +/- 2 degrees C on denitrification potential and respiration (determined as carbon dioxide evolution) were in general the same in moderately fertilized soils from four different sites: in all soils, depression of both the denitrification potential and potential respiration was found after 8 weeks. However, the extent to which the parameters were decreased differed from case to case. Not only the duration and storage conditions but also unidentified soil parameters are important for the persistence of biological activity in stored soils.

Cold Temperature↗

Vertical Distribution of Denitrification Potential, Denitrifying Bacteria, and Benzoate Utilization in Intertidal Microbial Mat Communities.

Aspects of denitrification and benzoate degradation were studied in two estuarine microbial mat communities on the California coast by measuring the depth distributions of potential denitrification rates, genetic potential for denitrification, nitrate concentration, benzoate mineralization rates, total bacterial abundance, and abundance of a denitrifying strain (TBD-8b) isolated from one of the sites. Potential denitrification was detected in microbial mat cores from both Elkhorn Slough and Tomales Bay. Maximum denitrification rates were more than two orders of magnitude higher at Elkhorn Slough (3.14 mmol N m -2 d -1) than at Tomales Bay (0.02 mmol N m -2 d -1), and at both sites, the maximum rates occurred in the 0-2 mm depth interval. Ambient pore [NO 3 + NO 2] was substantially higher at Elkhorn Slough than at Tomales Bay. Incorporation and mineralization of benzoate was maximal near the mat surface at Elkhorn Slough. The areal rate of benzoate utilization was 1045 nmol C m -2 d -1, which represented utilization of 70% of the added substrate in 24 h. Total bacterial and TBD-8b abundances were greatest near the surface at both Tomales Bay and Elkhorn Slough, and TBD-8b represented less than 0.2% of the total. Genetic potential for denitrification, quantified by hybridization with a nitrite reductase gene fragment, was present below the mat surface at average levels representing presence of the gene in approximately 10% of the total cells.

Journal Article↗

Monitoring the denitrification of wastewater containing high concentrations of nitrate with methanol in a sulfur-packed reactor.

Biological denitrification of high nitrate-containing wastewater was examined in a sulfur-packed column using a smaller amount of methanol than required stoichiometrically for heterotrophic denitrification. In the absence of methanol, the observed nitrate removal efficiency was only about 40%, and remained at 400 mg NO(3)(-)-N/l, which was due to an alkalinity deficiency of the pH buffer and of CO(2) as a carbon source. Complete denitrification was achieved by adding approximately 1.4 g methanol/g nitrate-nitrogen (NO(3)(-)-N) to a sulfur-packed reactor. As the methanol concentration increased, the overall nitrate removal efficiency increased. As influent methanol concentrations increased from 285 to 570, 855, and 1,140 mg/l, the value of Delta mg alkalinity as CaCO(3) consumed/Delta mg NO(3)(-)-N removed increased from -1.94 to -0.84, 0.24, and 0.96, and Delta mg SO(4)(2-) produced/Delta mg NO(3)(-)-N removed decreased from 4.42 to 3.57, 2.58, and 1.26, respectively. These results imply the co-occurrence of simultaneous autotrophic and heterotrophic denitrification. Sulfur-utilizing autotrophic denitrification in the presence of a small amount of methanol is very effective at decreasing both sulfate production and alkalinity consumption. Most of methanol added was removed completely in the effluent. A small amount of nitrite accumulated in the mixotrophic column, which was less than 20 mg NO(2)(-) -N/l, while under heterotrophic denitrification conditions, nitrite accumulated steadily and increased to 60 mg NO(2)(-) -N/l with increasing column height.

Bioreactors↗

Denitrification Distributions in Four Valley and Ridge RiparianEcosystems

/ Denitrification in riparian ecosystems can reduce the amount ofnitrogen transported from farm fields to streams. In this study, we examinedenitrification in four riparian ecosystems common to the Valley and Ridgephysiographic province in Pennsylvania, USA. The sites exhibit differentvegetation, are underlain by different rock types, and are downgradient offarm fields. Mean site denitrification rates ranging from 0.6 to 1.9 &mgr;gN/kg soil/day were measured using intact core incubation techniques. Thethree riparian sites covered with grass each exhibited greaterdenitrification rates than the wooded site. Denitrification rate wascorrelated with moisture content but not with nitrate-N or organic carboncontents. Denitrification rates were greatest near the soil surface and atpositions nearest the stream. Rates decreased uniformly with distance awayfrom the stream and also with depth in the soil for each site. While patternsof nitrate-N, moisture, and organic carbon content differ among the sites,their combined effects on denitrification support the observed, consistentdenitrification rate pattern.KEY WORDS: Denitrification; Riparian ecosystems

Journal Article↗

Monitoring microaerobic denitrification of Pseudomonas aeruginosa by online NAD(P)H fluorescence.

Defined as the transition conditions in which the organism(s) performs simultaneous aerobic and anaerobic respiration or fermentation, microaerobic conditions are commonly present in the nature. Microaerobic metabolism of microorganisms is however poorly characterized. Being extremely sensitive to the change in cellular electron-accepting mechanisms, NAD(P)H fluorescence provides a useful ways for online monitoring of microaerobic metabolism. Its application to studies of microbial nitrate respiration and particularly, denitrification of Pseudomonas aeruginosa is reviewed here, centering on four topics: (1) online monitoring of anaerobic nitrate respiration by NAD(P)H fluorescence, (2) effects of denitrification on P. aeruginosa phenotypes, (3) microaerobic denitrification of P. aeruginosa in continuous culture, and (4) correlation between NAD(P)H fluorescence and denitrification-to-respiration ratio. Online NAD(P)H fluorescence is shown to sensitively detect the changes of cellular metabolism. For example, it revealed the intermediate nitrite accumulation in C-limited Escherichia coli performing anaerobic nitrate respiration via dissimilative ammonification, by exhibiting two-stage profiles with intriguing fluorescence oscillation. When applied to continuous culture studies of P. aeruginosa (ATCC 9027), the online fluorescence helped to identify that the bacterium conducted denitrification even at DO > 1 mg/l. In addition, the fluorescence profile showed a unique correlation with the fraction of electrons accepted by denitrification (out of all the electrons accepted by aerobic and anaerobic respiration). The applicability of online NAD(P)H fluorescence in monitoring and quantitatively describing the sensitive microaerobic state of microorganisms is clearly demonstrated.

Aerobiosis↗

Nitrification-denitrification via nitrite accumulation for nitrogen removal from wastewaters.

The biological nitrification-denitrification process is used extensively for removal of ammonia nitrogen from wastewaters. Saves in aeration, organic matter (for denitrification) and surplus sludge are achievable if nitrite accumulation is possible in the nitrification step. In this paper, operational parameters were studied for each process for maximum nitrite accumulation in the nitrification step and nitrite adaptation in the denitrification step. Nitrite accumulation during nitrification can be controlled by the dissolved oxygen (DO) concentration, presenting a maximum of 65% at around 0.7 mg DO/L. Denitrification can be adapted to nitrite and the process is stable if nitrite in the reactor is keep low. The performance of a continuous stirred tank reactor (CSTR) and an up flow sludge blanket reactor (USB) were compared. Once the operational parameters were established, a CSTR for nitrification and an USB reactor for denitrification were operated in series for 25 days. The process was stable and a steady state was maintained for 20 days, and 93.5% of overall nitrogen removal was achieved in the nitrification-denitrification via the nitrite process.

Ammonia↗

Challenges for simultaneous nitrification, denitrification, and phosphorus removal in microbial aggregates: mass transfer limitation and nitrous oxide production.

The microbial community composition and activity was investigated in aggregates from a lab-scale bioreactor, in which nitrification, denitrification and phosphorus removal occurred simultaneously. The biomass was highly enriched for polyphosphate accumulating organisms facilitating complete removal of phosphorus from the bulk liquid; however, some inorganic nitrogen still remained at the end of the reactor cycle. This was ascribed to incomplete coupling of nitrification and denitrification causing NO(3)(-) accumulation. After 2 h of aeration, denitrification was dependent on the activity of nitrifying bacteria facilitating the formation of anoxic zones in the aggregates; hence, denitrification could not occur without simultaneous nitrification towards the end of the reactor cycle. Nitrous oxide was identified as a product of denitrification, when based on stored PHA as carbon source. This observation is of critical importance to the outlook of applying PHA-driven denitrification in activated sludge processes.

Bacteria, Aerobic↗

Macroscale and microscale analyses of nitrification and denitrification in biofilms attached on membrane aerated biofilm reactors.

A membrane aerated biofilm reactor (MABR), in which O(2) was supplied from the bottom of the biofilm and NH(4)(+) and organic carbon were supplied from the biofilm surface, was operated at different organic carbon loading rates and intra-membrane air pressures to investigate the occurrence of simultaneous chemical oxygen demand (COD) removal, nitrification and denitrification. The spatial distribution of nitrification and denitrification zones in the biofilms was measured with microelectrodes for O(2), NH(4)(+), NO(2)(-), NO(3)(-) and pH. When the MABR was operated at approximately 1.0 g-COD/m(2)/day of COD loading rate, simultaneous COD removal, nitrification and denitrification could be achieved. The COD loading rates and the intra-membrane air pressures applied in this study had no effect on the start-up and the maximum rates of NH(4)(+) oxidation in the MABRs. Microelectrode measurements showed that O(2) was supplied from the bottom of the MABR biofilm and penetrated the whole biofilm. Because the biofilm thickness increased during the operations, an anoxic layer developed in the upper parts of the mature biofilms while an oxic layer was restricted to the deeper parts of the biofilms. The development of the anoxic zones in the biofilms coincided with increase in the denitrification rates. Nitrification occurred in the zones from membrane surface to a point of ca. 60microm. Denitrification mainly occurred just above the nitrification zones. The COD loading rates and the intra-membrane air pressures applied in this study had no effect on location of the nitrification and denitrification zones.

Biofilms↗

Effect of salinity and inorganic nitrogen concentrations on nitrification and denitrification rates in intertidal sediments and rocky biofilms of the Douro River estuary, Portugal.

The regulatory effects of salinity and inorganic nitrogen compounds on nitrification and denitrification were studied in intertidal sandy sediments and rocky biofilms in the Douro River estuary, Portugal, over a 12-month period. Nitrification and denitrification rates were measured in slurries of field samples and enrichment experiments using the difluoromethane and the acetylene inhibition techniques, respectively. Salinity did not regulate denitrification in either environment, suggesting that halotolerant bacteria dominated the denitrifier communities. However, nitrification rates were stimulated when salinity increased from 0 to 15 practical salinity units. NO3- addition experiments revealed that NO3- availability stimulates denitrification rates in sandy sediments, but not in rocky biofilms; however, in rocky biofilms a positive and linear relationship was observed between denitrification rates and water column NO3- concentrations (r=0.92) during the monthly surveys. The N2O:N2 ratios increased rapidly when NO3- increased from 63 to 363 microM; however, results from monthly surveys showed that environmental parameters other than NO3- availability may be important in controlling the variation in N2O production via denitrification. Ammonium additions to sandy sediments stimulated nitrification rates by 35% for the 20 microM NH4+ addition, but NH4+ appeared to inhibit nitrification at high concentration addition (200 microM NH4+). In contrast, rocky biofilm nitrification was stimulated by 65% when 200 microM NH4+ was added.

Bacteria, Aerobic↗

Denitrification potential of industrial wastewaters.

Nitrate utilisation batch experiments have been carried out with different agro-food industrial wastewaters. The results obtained were modelled using a modification of the ASM2d to determine the main kinetic and stoichiometric parameters: denitrification rate with fermentation products (q(N-SA)) and with fermentable substrates (q(N-SF)); fermentation products and fermentable substrates consumption rates, (qSA) and (qSF), respectively; yield coefficient with fermentation products (Y(H-SA)) and with fermentable substrates (Y(H-SF)) and saturation coefficient for growth on fermentation products (K(H-SA)). It was found that fermentation products (SA) and fermentable substrates (SF) consumption rates can be assumed to be both constant and independent of the agro-food industrial wastewater used, with values of 14.6 and 5.4 (mg S(COD) g COD-X(-1)OHO h(-1)), respectively. The mean denitrification rates obtained with each agro-food industrial wastewater ranged from 1.5 to 2.5 (mg S(N) g COD-X(-1)OHO h(-1)), values slightly lower to the obtained with domestic wastewater (2.7 mg S(N) g COD-X(-1)OHO h(-1)). Given the substrate consumption rate and the denitrification rate, the yields with fermentation products and fermentable substrates for each wastewater were determined. The values obtained for the yields ranged from 0.50 to 0.69 on using SA and from 0.50 to 0.70 on using SF. Having determined these parameters, the denitrification potential (DNP) was evaluated for each agro-food wastewater. Wastewater from slaughterhouses gave the lowest DNP (22.6 mg S(N) g S(-1)COD), whereas wastewater from potato and tomato processing plants gave the highest values (126.5 and 128.2 mg S(N) g S(-1)COD), respectively), which are very similar to that obtained with domestic wastewater (130.3 and 128.2 mg S(N) g S(-1)COD). Taking into account the low N/COD ratio presented in the agro-food wastewaters, the last wastewaters are the most indicated to enhance the nitrogen removal in the WWTP. Finally, a relationship between the denitrification rates of several agro-food industrial wastewaters and their respective S(A)/(S(A) + S(F)) ratio was obtained. This relationship can be used to estimate the mean denitrification rate of an agro-food industrial wastewater.

Bioreactors↗

Denitrification potential and rates of complex carbon source from dairy effluents in activated sludge system.

A predictive model for the denitrification performance of complex carbon sources was proposed based on compositional data. Potential and rates of denitrification of single dairy components (lactose, lactate, proteins, fat), as well as binary and complex (modelled "process water") mixtures were assessed using test for nitrogen uptake rate (NUR). In all experiments, denitrification potential of mixtures was found to be significantly higher than the sum of individual potentials and denitrification rate with the readily biodegradable moiety of the mixtures was similar to the highest rate obtained with individual components (lactose or lactate). This work shows that activated sludge acclimated to dairy components can be modelled as a single biomass where the maximal anoxic growth rate of the biomass limits the denitrification rate with dairy components. As a consequence, lactose or lactate determine the maximal denitrification rate possible using dairy effluents.

Carbon↗

The effect of decreased ambient temperature on the biological nitrification and denitrification of a high ammonia landfill leachate.

This research investigated the nitrogen removal capabilities of a pre- and post-denitrification process, when treating sanitary landfill leachate containing an ammonia concentration of over 2200 mg N/L, at operating temperatures down to 10 degrees C. The treatment system employed is generally known as a 4-Stage Bardenpho process. Two parallel systems were operated throughout the study; each system had an aerobic solids retention time of 20 days and an external recycle ratio (return activated sludge) of about 3:1. During the entire experimental period, one system used an internal recycle ratio of about 4:1, while the other system used one of 3:1. Methanol was used as supplementary carbon source for denitrification. When operating temperature was decreased from 20 to 17 degrees C, an approximate 15% decrease in denitrification was immediately experienced within both systems, with no noticeable affect on nitrification. This was an unexpected result. Subsequently, the temperature was further decreased to 14 degrees C, and finally to 10 degrees C. With the temperature at 14 degrees C, aerobic wasting was also stopped and methanol loading was progressively decreased to match actual denitrification requirements. At 10 degrees C, both systems suffered major nitrification and denitrification inhibition. Changes in operating parameters, such as a decrease in influent ammonia and methanol loading, as well as an increase in ambient temperatures, from 10 to 15 degrees C, did not significantly improve overall system performance, within a reasonable time frame. Both denitrification and nitrification problems were mainly attributed to decreased ambient temperature, although other possible factors were noted; at project completion, about 50% of the incoming ammonia was being removed across the system.

Aerobiosis↗

Denitrification and neutralization treatment by direct feeding of an acidic wastewater containing copper ion and high-strength nitrate to a bio-electrochemical reactor process.

The feasibility of the direct denitrification treatment of copper metal pickling wastewater by using a bio-electrochemical reactor process was investigated experimentally. Carbon electrodes were installed in the reactor as the anode and cathode and denitrifying microorganisms were fixed on the surface of the cathode. The reactor was continuously operated by applying an electric current and feeding acetate. In this reactor, copper ion removal and denitrification proceeded simultaneously and the pH value of the treated water was increased almost to neutral. The electric current that passed through the cathode contributed to the removal of the copper ion and the generation of hydrogen gas. The generated hydrogen gas as well as the added acetate was effectively utilized for denitrification. A theoretical evaluation of pH in the effluent suggested that the pH increase was mainly caused by the generation of hydroxyl ion during denitrification. In addition, the inorganic carbon species generated during denitrification with acetate and by the electrochemical oxidation of anodic carbon acted as a buffer to minimize a further increase of pH at higher nitrate removal efficiencies. These results demonstrated that copper ion removal, denitrification and neutralization could be achieved simultaneously by using a single bioelectrochemical reactor.

Bioreactors↗

Simultaneous assessment of nitrification and denitrification on freshwater epilithic biofilms by acetylene block method.

Acetylene (C2H2) inhibits key enzymes involved in nitrification (Ammonium monooxygenase) and denitrification (N2O reductases). Thus an injection of C2H2 at mid time of a batch type incubation make it possible to assess denitrification by measurement of the N2O accumulation as well as nitrification, calculated from the variations of the ammonium flux. As estimated by the "acetylene block technique", denitrification is known to be only a measure of the denitrification rate supported by nitrate diffusing from the water column (Dw). This paper presents a first application on river epilithic biofilms which proved that the simultaneous measurement of Dw and nitrification allows the estimation of the order of magnitude of total denitrification (Dt) when nitrification is detected in the tested sample. This approach appears to be an easy tool for determination of nitrification and denitrification in natural samples and as thus presents an alternative to isotopic 15N methods.

Acetylene↗

Biological nitrogen removal from municipal landfill leachate: low-cost nitrification in biofilters and laboratory scale in-situ denitrification.

The slow leaching of nitrogen from solid waste in landfills, resulting in high concentrations of ammonia in the landfill leachate, may last for several decades. The removal of nitrogen from leachate is desirable as nitrogen can trigger eutrophication in lakes and rivers. In the present study, a low-cost nitrification-denitrification process was developed to reduce nitrogen load especially in leachates from small landfills. Nitrification was studied in laboratory and on-site pilot aerobic biofilters with waste materials as filter media (crushed brick in upflow filters and bulking agent of compost in a downflow filter) while denitrification was studied in a laboratory anoxic/anaerobic column filled with landfill waste. In the laboratory nitrification filters, start-up of nitrification took less than 3 weeks and over 90% nitrification of leachate (NH4-N between 60 and 170mg N l(-1), COD between 230 and 1,300 mg l(-1)) was obtained with loading rates between 100 and 130 mgNH4-N l(-1) d at 25 degrees C. In an on-site pilot study a level of nitrification of leachate (NH4-N between 160 and 270 mg N l(-1), COD between 1,300 and 1,600 mg l(-1)) above 90% was achieved in a crushed brick biofilter with a loading rate of 50mg NH4-N l(-1) d even at temperatures as low as 5-10 degrees C. Ammonium concentrations in all biofilter effluents were usually below the detection limit. In the denitrification column. denitrification started within 2 weeks and total oxidised nitrogen in nitrified leachate (TON between 50 and 150mg N l(-1)) usually declined below the detection limit at 25 degrees C, whereas some ammonium, probably originating from the landfill waste used in the column, was detected in the effluent. No adverse effect was observed on the methanation of waste in the denitrification column with a loading rate of 3.8 g TON-N/t-TS(waste) d. In conclusion, nitrification in a low-cost biofilter followed by denitrification in a landfill body appears applicable for the removal of nitrogen in landfill leachate in colder climates.

Biofilms↗

Factors influencing deterioration of denitrification by oxygen entering an anoxic reactor through the surface.

The purpose of the paper is to examine the factors that influence the deterioration of denitrification in open anoxic reactors. For this investigation an ASM 1-based simulation model was developed and successfully applied to fit data from batch experiments carried out in lab-scale reactor vessels (uncovered and covered) using both clarified domestic wastewater and synthetic wastewater. Applying the verified model, simulation studies were performed to investigate the effects of available denitrifiable substrate, biomass concentration, oxygen transfer rate, and temperature on deterioration of denitrification in open anoxic reactors. It has been shown that oxygen entering an anoxic reactor through the surface may not just affect denitrification metabolically, but also kinetically, due to increased dissolved oxygen (DO) concentration exerting an inhibitory effect on the denitrification rate. When the exogenous substrate concentration in the reactor vessel is high enough for a high consumption rate, the DO concentration is kept low. The higher the biomass concentration, and thereby the consumption rate of endogenous substrate, the lower the DO concentration during the low-rate denitrification phase. At low substrate removal rates, decreasing temperature will cause the DO concentration in anoxic vessels to increase. The results suggest that assuring removal of available exogenous carbon source at high rate by staging of open anoxic bioreactors may significantly improve denitrification efficiency.

Biomass↗

Kinetics of sequential nitrification and denitrification processes.

Kinetics of nitrification and denitrification of synthetic wastewater was investigated by using two reactors in series. An activated sludge unit was used for nitrification followed by a downflow biofilter for denitrification. Glucose solution was fed to the denitrification column to supply carbon source. Reactors were operated at different operating conditions and data were collected for determination of kinetic constants. Experimental data indicated that nitrification and denitrification kinetics followed Monod kinetics. By using the experimental data, kinetic constants for nitrification were determined as k = 1.15 d(-1), K(N) = 5.14 mg/l, Y = 0.34 mgX/mgN and b = -0.021 d(-1). Similarly, kinetic constants for denitrification were determined as k = 0.23 d(-1) and K(DN) = 0.27 mg/l. Rates of nitrification and denitrification increased with increasing nitrogen loading rate.

Journal Article↗