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Nitrous oxide nitrification and denitrification 15N enrichment factors from Amazon forest soils.

The isotopic signatures of 15N and 18O in N2O emitted from tropical soils vary both spatially and temporally, leading to large uncertainty in the overall tropical source signature and thereby limiting the utility of isotopes in constraining the global N2O budget. Determining the reasons for spatial and temporal variations in isotope signatures requires that we know the isotope enrichment factors for nitrification and denitrification, the two processes that produce N2O in soils. We have devised a method for measuring these enrichment factors using soil incubation experiments and report results from this method for three rain forest soils collected in the Brazilian Amazon: soil with differing sand and clay content from the Tapajos National Forest (TNF) near Santarém, Pará, and Nova Vida Farm, Rondônia. The 15N enrichment factors for nitrification and denitrification differ with soil texture and site: -111 per thousand +/- 12 per thousand and -31 per thousand +/- 11 per thousand for a clay-rich Oxisol (TNF), -102 per thousand +/- 5 per thousand and -45 per thousand +/- 5 per thousand for a sandier Ultisol (TNF), and -10.4 per thousand +/- 3.5 per thousand (enrichment factor for denitrification) for another Ultisol (Nova Vida) soil, respectively. We also show that the isotopomer site preference (delta15Nalpha - delta15Nbeta, where alpha indicates the central nitrogen atom and beta the terminal nitrogen atom in N2O) may allow differentiation between processes of production and consumption of N2O and can potentially be used to determine the contributions of nitrification and denitrification. The site preferences for nitrification and denitrification from the TNF-Ultisol incubated soils are: 4.2 per thousand +/- 8.4 per thousand and 31.6 per thousand +/- 8.1 per thousand, respectively. Thus, nitrifying and denitrifying bacteria populations under the conditions of our study exhibit significantly different 15N site preference fingerprints. Our data set strongly suggests that N2O isotopomers can be used in concert with traditional N2O stable isotope measurements as constraints to differentiate microbial N2O processes in soil and will contribute to interpretations of the isotopic site preference N2O values found in the free troposphere.

Brazil↗

Denitrification at a long-term forested land treatment system in the Piedmont of Georgia.

Spray irrigation of forested land can provide an effective system for nutrient removal and treatment of municipal wastewater. Evolution of N2 + N2O from denitrifying activity is an important renovation pathway for N applied to forested land treatment systems. Federal and state guidance documents for design of forested land treatment systems indicate the expected range for denitrification to be up to 25% of applied N, and most forest land treatment systems are designed using values from 15 to 20% of applied N. However, few measurements of denitrification following long-term wastewater applications at forested land treatment sites exist. In this study, soil N2 + N2O-N evolution was directly measured at four different landscape positions (hilltop, midslope, toe-slope, and riparian zone) in a forested land treatment facility in the Georgia Piedmont that has been operating for more than 13 yr. Denitrification rates within effluent-irrigated areas were significantly greater than rates in adjacent nonirrigated buffer zones. Rates of N2 + N2O-N evolved from soil in irrigated forests ranged from 5 to 10 kg ha(-1) yr(-1) N on the three upland landscape positions and averaged 38 kg ha(-1) yr(-1) N within the riparian zone. The relationship between measured riparian zone denitrification rates and soil physical and chemical properties was poor. The best relationship was with soil temperature, with an r2 of 0.18. Overall, on a landscape position weighted basis, only 2.4% of the wastewater-applied N was lost through denitrification.

Ecosystem↗

Nitrification and denitrification rates of Everglades wetland soils along a phosphorus-impacted gradient.

Little information is available on the effect of phosphorus (P) enrichment on nitrogen (N) biogeochemical cycling in wetland soil. Of particular importance are the coupled nitrification-denitrification reactions that regulate the microbially mediated loss of N from wetland systems. Soils from the northern Florida Everglades have been affected by P loading from surface waters over the past 40 years. Elevated P levels have been show to have an effect on the size and activity of the microbial pool and a decrease in the N to P ratio of the microbial biomass. The objective of the study was to determine if P enrichment in soils affected microbial activities related to nitrification and denitrification in these flooded, peat soils. Potential nitrification rates of soil and detritus were determined using constantly stirred reactors under aerobic conditions while denitrification rates were determined from anaerobic incubations of slurry. Nitrification rates showed two distinct linear phases, a slower initial rate, signifying activity of nitrifiers present, followed by a sharp increase in the NH4+ conversion rate indicative of maximum potential rates. Initial rates of nitrification were highest in the surficial detrital layer decreasing with soil depth and did not correlate to soil total P. The potential rates of nitrification were 13 times greater than the initial rates. Potential denitrification rates were highest in the detritus and 0- to 10-cm soil interval with significantly lower values in the 10- to 30-cm soil interval, significantly correlated to total P of the soil. A significant (P < 0.01) relationship was seen between potential denitrification rates and soil total P suggesting an increased rate of N removal from P-enriched regions of the northern Everglades.

Conservation of Natural Resources↗

Denitrification as a nitrogen removal mechanism in a Vermont peatland.

Atmospheric deposition of nitrogenous compounds to ombrotrophic peatlands (i.e., those that have peat layers higher than their surroundings and receive nutrients and minerals exclusively by precipitation) has the potential to significantly alter ecosystem functioning. This study utilized the acetylene inhibition technique to estimate the relative importance of denitrification in nitrogen removal from a primarily ombrotrophic peatland, in an attempt to estimate the threat of increased nitrogen loadings to these areas. Estimates of mean rates of denitrification ranged from -2.76 to 84.0 ng N(2)O-N cm(-3) h(-1) (equivalent to -150 to 4800 microg N(2)O-N m(-2) h(-1)) using an ex situ core technique and from -8.30 to 5.98 microg N(2)O-N m(-2) h(-1) using an in situ chamber technique. Core rates may have been elevated over natural field levels due to effects of disturbance on substrate availability, and chamber rates may have been low due to diffusional constraints on acetylene and N(2)O. Net nitrification was also measured in an attempt to evaluate this process as a source of nitrate for denitrifiers. The low rates of net nitrification measured, in combination with the low rates of in situ denitrification and the very low amounts of free nitrate measured in this peatland, suggests that inorganic N turnover in this wetland is low. Results showed that nitrate was a limiting factor for denitrification in this peatland, with mean rates from nitrate-amended cores ranging from 13.1 to 260 ng N(2)O-N cm(-3) h(-1), and it is expected that increases in nitrogen loadings will increase denitrification rates in this ecosystem.

Acetylene↗

Comparison of nutrient removal efficiency between pre- and post-denitrification wastewater treatments.

A shortage of organic substances (COD) may cause problems for biological nutrient removal, that is, lower influent COD concentration leads to lower nutrient removal rates. Biological phosphorus removal and denitrification are reactions in which COD is indispensable. As for biological simultaneous nitrogen and phosphorus removal systems, a competition problem of COD utilisation between polyphosphate accumulating organisms (PAOs) and non-polyphosphate-accumulating denitrifiers is not avoided. From the viewpoint of effective utilisation of limited influent COD, denitrifying phosphorus-removing organisms (DN-PAOs) can be effective. In this study, DN-PAOs activities in modified UCT (pre-denitrification process) and DEPHANOX (post-denitrification process) wastewater treatments were compared. In conclusion, the post-denitrification systems can use influent COD more effectively and have higher nutrient removal efficiencies than the conventional pre-denitrification systems.

Aerobiosis↗

Nitrogen removal from the saline sludge liquor by electrochemical denitrification.

Sludge liquor from the sludge dewatering process has a high ammonia content. In the present study, a lab-scale electrochemical (EC) system with a pair of Ti electrode plates was used for treating the sludge centrate liquor of digested wastewater sludge with a NH4(+) - N content of around 500 mg/L. The sludge liquor had a high salinity due to seawater being used for toilet flushing in Hong Kong. The results show that the EC process is highly effective for denitrification of the saline sludge liquor. Complete nitrogen removal could be achieved within 1 hr or so. The rate of EC denitrification increased with the current intensity applied. The best current efficiency for nitrogen removal was obtained for a gap distance between the electrodes at 8 mm. Electro-chlorination was considered to be the major mechanism of EC denitrification. The formation of chlorination by-products (CBPs) appeared to be minimal with the total trihalomethanes (THM) detected at a level of 300 microg/L or lower. The power consumption for EC denitrification was around 23 kWh/kg N. Additional electro-flocculation with a pair of iron needle electrodes could enhance the flocculation and subsequent sedimentation of colloidal organics in the sludge liquor, increasing the organic removal from less than 30% to more than 70%. Therefore, the EC process including both electro-denitrification and electro-flocculation can be developed as the most cost-effective method for treatment of the saline sludge liquor.

Bioreactors↗

Biological denitrification of hydrolysates from octahydro-1,3,5,7 tetranitro-1,3,5,7-tetrazocine.

Alternatives for the destruction of common military explosives, including trinitrotoluene; hexahydro-1,3,5-trinitro-1,3,5-triazine; and octahydro-1,3,5,7 tetranitro-1,3,5,7-tetrazocine (HMX) are being investigated in the post-cold war period. One alternative combines chemical treatment (i.e., base hydrolysis of the explosives) and biological treatment (i.e., denitrification of the hydrolysate). This paper focuses on results of the biological part of the treatment process, during which Hyphomicrobium sp. bacteria were isolated from a seed obtained from a denitrification facility. The bacteria were enriched and maintained on a surrogate waste with methanol as the carbon source. The resulting culture is capable of anoxic growth in waste solutions containing up to 5000 mg/L of nitrite-nitrogen. The culture efficiently denitrifies both surrogate and actual hydrolysate wastes. A substrate inhibition model was used to accurately predict denitrification rates. Comparisons are made between denitrification rates obtained for surrogate versus actual wastes. Denitrification rates were higher when actual waste streams were used. This work demonstrates the feasibility of using Hyphomicrobiun sp. bacteria to treat HMX hydrolysate and presents a model that can be used to design a large-scale system.

Azocines↗

Changes in anoxic denitrification rate resulting from prefermentation of a septic, phosphorus-limited wastewater.

A preliminary bench-scale study of parallel University of Cape Town (UCT) biological nutrient removal systems showed improvement in anoxic denitrification rates resulting from prefermentation of a septic (i.e., high volatile fatty acid [VFA] content), phosphorus-limited (i.e., total chemical oxygen demand/total phosphorus [TP] ratio < 40:1) wastewater. Net phosphorus removals due to enhanced biological phosphorus removal (EBPR) were only improved marginally by prefermentation in spite of significant increases in anaerobic phosphorus release, polyhydroxyalkanoate formation, and higher anoxic and aerobic uptakes. This probably was due to the high VFA/TP ratio in the raw influent relative to the VFA requirements for EBPR because enough VFAs were already present for phosphorus removal prior to prefermentation. An additional assessment of prefermentation using parallel UCT systems with step feed of 50% of the influent to the anoxic zone was completed. This second phase quantified the effect of prefermentation in a step-feed scenario, which prioritized prefermentation use to enhance denitrification rather than EBPR. While specific denitrification rates in the anoxic zone were significantly improved by prefermentation, high denitrification in the clarifiers and aerobic zones (simultaneous denitrification) made definitive conclusions concerning the potential improvements in total system nitrogen removal questionable. The prefermented system always showed superior values of the zone settling velocity and sludge volume index and the improvement became increasingly statistically significant when the prefermenter was performing well.

Bacteria, Anaerobic↗

Autotrophic denitrification of high-salinity wastewater using elemental sulfur: batch tests.

The feasibility of autotrophic denitrification of a high-salinity wastewater using sulfur-oxidizing autotrophic denitrificants was studied. These autotrophic bacteria oxidize elemental sulfur to sulfate while reducing nitrate to elemental nitrogen gas, thereby eliminating the need for the addition of organic carbon compounds. A series of bench-scale batch tests was performed with synthetic and actual flue gas desulfurization (FGD) wastewater to examine the effects of various environmental and operational factors such as temperature, pH, high salinity, and potentially toxic substances on the rate of autotrophic denitrification using elemental sulfur. Specific denitrification rates of 6 to 8 mg nitrate-nitrogen (NO3- -N)/g volatile suspended solids (VSS) x h were obtained. The highest denitrification rates were found between pH 7.0 and 8.0 and a temperature of 30 degrees C. The denitrification rate started to decrease above an osmotic pressure of 19 atm (approximately 70% of seawater), independent of the type of salt ions, and amounted to approximately 70% activity at the concentration of seawater. Polyphosphate or pyrophosphate could be used as the source of phosphorus instead of orthophosphate because the latter caused immediate calcium phosphate precipitation in the FGD wastewater. Inhibiting factors attributed to inorganic or organic compounds originating from coal combustion were discussed.

Hydrogen-Ion Concentration↗

[Isolation of bacteria which could perform nitrification and denitrification simultaneously by gene probe].

The bacteria which could perform nitrification and denitrification simultaneously from nitrogen containing wastes in Taiwan were isolated by using the probes made from random DNA fragments of Thiosphaera pantotropha. Two isolates were identified and named Alcaligenes faecalis subsp. faecalis strain 1 and strain 2 respectively. The effects on nitrification and denitrification by different medium pH, oxygen content, addition of different electron donors or inhibitors were studied. The isolates not only could perform nitrification, but also denitrification even in the presence of oxygen. Potassium cyanide could inhibit denitrification; hydrazine and hydroxyamine could inhibit nitrification. Alcaligenes faecalis subsp. faecalis strain 2 shows better denitrification.

Alcaligenes↗

Denitrification potential enhancement by addition of anaerobic fermentation products from the organic fraction of municipal solid waste.

The aim of this study is to demonstrate the denitrification potential enhancement determined by the addition of the anaerobic fermentation products from the organic fraction of municipal solid waste in BNR wastewater treatment plants. The denitrification potential rose from 6 to 17 mgNO3-N/l thanks to the good performances showed by the fermentation products in the denitrification step: a typical maximum denitrification rate of about 5 mgN/gVSS d was observed. This value is similar to those obtained by using pure organic substrates. The denitrification capacity increase is determined by a significant increase of S8 and Xs availability in the influent due to the anaerobic fermentation products addition which leads to an influent COD increase equal to 68% of the total load: this value is significantly higher than the one obtained by fermentation of primary and/or waste activated sludge (3-5%).

Bacteria, Anaerobic↗

Simultaneous nitrification and denitrification in a single reactor using bio-electrochemical process.

Feasibility of a bio-electrochemical process for simultaneous nitrification and denitrification in a single reactor was experimentally investigated. The reactor consisted of anodic and cathodic electrodes, on which nitrifying and denitrifying biofilms, respectively, were fixed. Effects of the applied electric current and DO concentration in the bulk solution were examined. The TN removal could be achieved through the occurrence of nitrification and denitrification in anodic and cathodic biofilms, respectively. Both nitrification and denitrification rates increased with an increase in the applied electric current. Even at low DO concentration in the bulk solution, nitrification proceeded at a high rate by utilizing oxygen generated on the anode. Denitrification rate remained relatively high at high DO concentration due to supplying hydrogen gas to the inner side of the cathodic biofilm. The higher TN removal rate tended to be obtained at lower DO concentration and higher current density. From these results, it was concluded that the bio-electrochemical process was applicable to simultaneous nitrification and denitrification due to stable formation of aerobic and anoxic regions in the single reactor.

Bacteria, Aerobic↗

Nitrate removal by simultaneous sulfur utilizing autotrophic and heterotrophic denitrification under different organics and alkalinity conditions: batch experiments.

The effect of various organic compounds were tested using lab-scale batch reactors. At sufficient alkalinity, the initial nitrate nitrogen concentration of 100 mg/L was completely reduced in all batch reactors. Sulfate production decreased by the addition of organics. The concentration range of organics used in this experiment did not inhibit autotrophic denitrification except for propionate. Propionate inhibited autotrophic denitrification a little, indicated by a lower sulfate production rate. Biomass in suspension increased with higher initial organic concentrations, showing higher DOC consumption. As the concentration of organics increased, alkalinity increased accordingly. Under the conditions of low alkalinity, in the case of a control reactor without organics, only about 30% of the initial nitrate was reduced. With half the theoretically required dosage of methanol, the denitrification rates increased slightly. When ethanol, acetate, and propionate were used, denitrification went to completion. When excess organics was added, however, sulfate production was significantly decreased. Interestingly, even when small amounts of organics were added, autotrophic denitrification was promoted as indicated by the sulfate production.

Bioreactors↗

Denitrification in constructed wetlands used for treatment of swine wastewater.

Constructed wetland treatment of swine wastewater probably involves substantial denitrification. Our objective was to assess denitrification and denitrification enzyme activity (DEA) in such wetlands in relation to plant communities, N loading, carbon or nitrogen limitations, and water depth. Two wetland cells each 3.6 m wide and 33.5 m long were connected in series. One set of cells was planted with rushes and bulrushes, including soft rush (Juncus effusus L.), softstem bulrush [Schoenoplectus tabernaemontani (K.C. Gmel.) Pallal, American bulrush [Schoenoplectus americanus (Pers.) Volkart ex Schinz & R. Keller], and woolgrass bulrush [Scirpus cyperinus (L.) Kunth]. Another set was planted with bur-reeds and cattails, including American bur-reed (Sparganium americanum Nutt.), broadleaf cattail (Typha latifolia L.), and narrowleaf cattail (Typha angustifolia L.). The sets will be referred to herein as bulrush and cattail wetlands, respectively. Denitrification and DEA were measured via the acetylene inhibition method in intact soil cores and disturbed soil samples that were taken during four years (1994-1997). Although DEA in the disturbed samples was greater than denitrification in the core samples, the measurements were highly correlated (r2 > or = 0.82). The DEA was greater in the bulrush wetlands than the cattail wetlands, 0.516 and 0.210 mg N kg(-1) soil h(-1), respectively; and it increased with the cumulative applied N. The DEA mean was equivalent to 9.55 kg N ha(-1) d(-1) in the bulrush wetlands. We hypothesized and confirmed that DEA was generally limited by nitrate rather than carbon. Moreover, we determined that one of the most influential factors in DEA was wetland water depth. In bulrush wetlands, the slope and r2 values of the control treatment were -0.013 mg N kg(-1) soil h(-1) mm(-1) depth and r2 = 0.89, respectively. Results of this investigation indicate that DEA can be very significant in constructed wetlands used to treat swine wastewater.

Animals↗

A novel wastewater treatment process: simultaneous nitrification, denitrification and phosphorus removal.

Simultaneous nitrification and denitrification (SND) via the nitrite pathway and anaerobic-anoxic enhanced biological phosphorus removal (EBPR) are two processes that can significantly reduce the COD demand for nitrogen and phosphorus removal. The combination of these two processes has the potential of achieving simultaneous nitrogen and phosphorus removal with a minimal requirement for COD. A lab-scale sequencing batch reactor (SBR) was operated in alternating anaerobic-aerobic mode with a low dissolved oxygen concentration (DO, 0.5 mg/L) during the aerobic period, and was demonstrated to accomplish nitrification, denitrification and phosphorus removal. Under anaerobic conditions, COD was taken up and converted to polyhydroxyalkanoates (PHA), accompanied with phosphorus release. In the subsequent aerobic stage, PHA was oxidized and phosphorus was taken up to less than 0.5 mg/L at the end of the cycle. Ammonia was also oxidised during the aerobic period, but without accumulation of nitrite or nitrate in the system, indicating the occurrence of simultaneous nitrification and denitrification. However, off-gas analysis found that the final denitrification product was mainly nitrous oxide (N2O) not N2. Further experimental results demonstrated that nitrogen removal was via nitrite, not nitrate. These experiments also showed that denitrifying glycogen-accumulating organisms rather than denitrifying polyphosphate-accumulating organisms were responsible for the denitrification activity.

Acetates↗

The presence of ammonium facilitates nitrite reduction under PHB driven simultaneous nitrification and denitrification.

For economic and efficient nitrogen removal from wastewater treatment plants via simultaneous nitrification and denitrification the nitrification process should stop at the level of nitrite such that nitrite rather than nitrate becomes the substrate for denitrification. This study aims to contribute to the understanding of the conditions that are necessary to improve nitrite reduction over nitrite oxidation. Laboratory sequencing batch reactors (SBRs) were operated with synthetic wastewater containing acetate as COD and ammonium as the nitrogen source. Computer controlled operation of the reactors allowed reproducible simultaneous nitrification and denitrification (SND). The oxygen supply was kept precisely at a low level of 0.5 mgL(-1) and bacterial PHB was the only electron donor available for denitrification. During SND little nitrite or nitrate accumulated (< 20% total N), indicating that the reducing processes were almost as fast as the production of nitrite and nitrate from nitrification. Nitrite spiking tests were performed to investigate the fate of nitrite under different oxidation (0.1-1.5 mgL(-1) of dissolved oxygen) and reduction conditions. High levels of reducing power were provided by allowing the cells to build up to 2.5 mM of PHB. Nitrite added was preferentially oxidised to nitrate rather than reduced even when dissolved oxygen was low and reducing power (PHB) was excessively high. However, the presence of ammonium enabled significant reduction of nitrite under low oxygen conditions. This is consistent with previous observations in SBR where aerobic nitrite and nitrate reduction occurred only as long as ammonium was present. As soon as ammonium was depleted, the rate of denitrification decreased significantly. The significance of the observed strongly stimulating effect of ammonium on nitrite reduction under SND conditions is discussed and potential consequences for SBR operation are suggested.

Acetates↗

Integration of titrimetric measurement, off-gas analysis and NOx- biosensors to investigate the complexity of denitrification processes.

The denitrification process, namely the reduction of nitrate (NO3-) to nitrogen gas (N2), often cannot be simply modelled as a single step process. For a more complete and comprehensive model the intermediates, particularly nitrite (NO2-) and nitrous oxide (N2O), need to be investigated. This paper demonstrates the integration of titrimetric measurements and off-gas analysis with on-line nitrite plus nitrate (NOx-) biosensors, highlighting the necessity of measuring process intermediates with high time-scale resolution to study and understand the kinetics of denitrification. Investigation of activated sludge from a full-scale treatment plant showed a significant accumulation of NO2-, which appeared to impact on the overall denitrification rate measured as NOx- reduction or N2 production. A different sludge obtained from a lab-scale bioreactor produced N2O instead of N2 as the end product of denitrification. The two examples both illustrate the complexity of denitrification and stress the need for the more versatile and detailed measurement procedures, as presented in this paper.

Bioreactors↗

[Potential of nitrification and denitrification in water purification system with hydroponic bio-filter method].

The potential of nitrification and denitrification of sediment and the density of ammonium-oxidizing bacteria and nitrite-oxidizing bacteria in sediment in water quality purifying system with hydroponic bio-filter method (HBFM) were measured. The variation of nitrification and denitrification potential of the sediment along the stream way was quantitatively studied. The results show that among the sediments from front, middle and retral part of the stream way, the sediment from middle part reached a maximum nitrification potential . nitrification potential of 4.76 x 10(-6) g/(g x h), while the sediment from front part reached a maximum denitrification potential of 8 .1 x 10(-7) g/(g x h). The distribution of nitrification potential accords with the ammonium-oxidizing bacteria density. The key for improving nitrogen removal efficiency of HBFM system consists in changing nitrification & denitrification region distributing and accordingly enhances denitrification process.

Biofilms↗