Wetland-based treatment systems are often implemented as a method to remove unwanted substances from contaminated groundwater. Wetlands are effective due to the high biological activity that naturally takes place in the rhizosphere and soil. In support of a demonstration surface wetland system at a site in Columbus, Georgia, laboratory-scale wetland systems were designed to study the effect of different carbon sources and their biodegradability, COD:N ratio and temperature on the rate and extent of nitrate reduction of nitrate-bearing groundwater. Nitrate reducing bacteria are ubiquitous in surface and subsurface wetlands but a major limiting factor for these systems is carbon availability. Two major carbon sources were investigated in both continuous-flow and batch systems: a natural source, hay and a commercial source, MicroC GTM, a concentrated carbohydrate mix. Between these two carbon sources, the nitrate removal rate was not significantly different as long as sufficient biodegradable carbon was provided. The effect of both hydraulic retention time (HRT) and COD:N ratio on nitrate removal were investigated in continuous-flow systems. The specific nitrate removal rate in open to the atmosphere batch reactors was estimated at 0.55 mg N/mg biomass VSS-day. The effluent nitrate concentration in a continuous-flow system maintained with an HRT of 5 days at room temperature (22 to 23°C) was less than 3 mg nitrate-N/L. The COD:N ratio was kept at 6:1 for the majority of the experiments (approximately twice the theoretical requirement) to ensure sufficient carbon loading. Lower COD:N ratios of 5, 4, 3, 2, 1, and 0.5 were also investigated in the continuous-flow system and the minimum required carbon loading to achieve an effluent nitrate concentration below 10 mg N/L for an influent groundwater nitrate concentration between 65 and 70 mg N/L was determined to be 5:1 COD:N. The effect of temperature on the nitrate removal rate was also investigated at 22, 15, 10 and 5°C. As expected, the rate of nitrate reduction decreased with the decrease in temperature, especially below 10°C. Overall, the surface wetland is a feasible solution to treating nitrate-bearing groundwater even at relatively low ambient temperature values, provided that sufficient, biodegradable carbon is present.

This research estimates nitrogen removal from stormwater runoff using a denitrifying bioretention system using the USEPA Storm Water Management Model Version 5 (SWMM-5). SWMM-5 has been used to help planners make better decisions since its development in 1971. A conventional bioretention system is a type of Low Impact Development (LID) technology, which designed without a media layer specifically for achieving nitrogen removal. More recently studies have showed that high TN removal efficiencies are possible when incorporating a denitrification media layer. These systems are known as denitrifying bioretention systems, or alternative bioretention systems. LID projects are currently being designed and developed in Sarasota County, Florida. These projects include a bioretention cell retrofit project on Venice East Blvd., in Venice, FL where thirteen bioretention cells will be developed. Although implementation of LID has already begun in southwest Florida, little research exists on whether these systems are effective at reducing non-point sources of nutrients. Therefore, the overall goal of this research project was to investigate the performance of a proposed bioretention system in Venice, FL to treat non-point sources of nitrogen from stormwater runoff. An alternative bioretention cell (ABC) model was designed to conceptually address water routing through a layered bioretention cell by separating the model into treatment layers- the layers where the nitrification and denitrification reactions are expected to occur within an alternative bioretention system (i.e., nitrification is assumed to occur in the sand media layer, and denitrification in the wood chip media layer). The bioretention cell configuration was based largely on the development plans provided by Sarasota County; however, the configuration incorporated the same electron donor media for denitrification that was used in a prior study (i.e., wood chips). Site-specific input parameters needed to calibrate the ABC model were obtained from laboratory analyses, the literature, and the US Geological website (websoilsurvey.nrcs.usda.gov). Using a mass balance approach, and the hydraulic residence time (HRT) values from the results of a previous study, first-order loss rate coefficients for both nitrification and denitrification (k1 and k2, respectively) were estimated. The rate coefficients were then used to develop treatment expression for nitrification and denitrification reactions. The treatment expressions were used to estimate the annual load reductions for TKN, NO3--N, and TN at the Venice East Blvd. bioretention retrofit site. Six storm events were simulated using a range of nitrogen concentrations. The simulation results showed minimal nitrification removal rates for storm events exceeding 1 inch, due to the planned bioretention system area being only 1% of the subcatchment area. A new ABC model was created (based on EPA bioretention cell sizing guidelines), to be 6% of the subcatchment area. Both systems were used to estimate TN removal efficiencies. The larger sized ABC model results showed average TKN, NO3--N and TN reductions of 84%, 96%; and 87%, respectively; these are comparable to results from similar studies. Results indicate that adequate nitrogen attenuation is achievable in the alternative bioretention system, if it is sized according to EPA sizing guidelines (5-7%).

Ocean nitrogen fixation and denitrification are crucial nitrogen source and sink mechanisms for the global ocean environment. While recent studies have reported that oceanic denitrification has increased over the last few decades, others have suggested that global ocean nitrogen fixation rates have been underestimated, and still others that anthropogenic perturbations have altered the global nitrogen cycle. This implies that the current estimates of the oceanic nitrogen inventory are incomplete and they need to be revised with more information. In addition, current denitrification estimates need to be reexamined due to their large associated uncertainties. Thus, I have conducted research estimating denitrification rates in three different locations: the northern Gulf of Mexico (GOM), the East/Japan Sea (EJS), and the South Pacific: from coastal to marginal to open ocean scale in different oceanographic conditions. Denitrification rates in the bottom layer (including bottom waters+sediments) at the shallow and often hypoxic northern GOM ranged from 103-544 [mu]mol N m⁻² d⁻¹ (=1.4 to 7.4 Gg N mon⁻¹ with area=3.24x10¹⁰m²), and were controlled not only by biogeochemical factors (i.e. organic matter supply and remineralization), but also by physical factors (i.e. stratification and relative contributions from different water masses). Despite high dissolved oxygen concentrations, the significant decrease in nitrate concentrations below the expected levels, low N/P ratio (<12.4), and deep nitrite peak in the bottom layer indicate a presence of denitrification in EJS, confined at the Tatar Strait and the Ulleung Basin areas. The estimated denitrification rates range from 0.3 to 33.2 [mu]mol N m⁻² d⁻¹, and was comparable to the directly measured denitrification rates from sediment samples. The high-quality repeat hydrographic datasets observed at 32°S of the South Pacific Ocean offer an opportunity to estimate water column denitrification rates on a basin-scale in the open ocean away from the Eastern Tropical Pacific oxygen minimum zones. The mean water column denitrification rates in the oxygen minimum layer of P06 line (32°S) were estimated to range between 7.1 and 18.5 [mu]mol N m⁻² d⁻¹. The results imply that, although very small at any particular site, once integrated over a basin-scale, the open ocean water column denitrification can be a significant component of the oceanic nitrogen budget. Denitrification is subject to seasonal, decadal and possibly climate scale variations. While it is commonly estimated at the oxygen minimum zones or sediments, denitrification is not merely confined to such regions only, and small amounts of denitrification occur in other oceanic parts. Once integrated, it may be quantitatively significant for the world's oceans. Denitrification is playing a significant role in local, regional, and global ocean scales. In the future, we need to consider variability of denitrification in coastal regions, and to investigate denitrification in unexpected and unexplored regions, in order to improve our knowledge on global oceanic mass balance. / text

In natural waters, high concentrations of ammonia are toxic to fish, and the oxidation of ammonia to nitrate (NO₃-) consumes large quantities of dissolved oxygen. The influent to municipal wastewater treatment plants in the United States typically contains approximately 40 mg/L of ammonia nitrogen (NH₃₋ N). Almost all of this ammonia must be removed in a wastewater treatment process before the effluent is discharged to the natural environment. This dramatic decrease is accomplished by the aerobic biological process of nitrification, in which ammonia is oxidized to nitrate Biological denitrification is an anoxic biological process in which nitrate (NO₃-) is reduced to nitrogen gas (N₂). Denitrification can increase the alkalinity in activated sludge aeration basins and decrease the concentration of filamentous organisms. The staff at the City of Austin Water Utility decided to implement a denitrification system at Walnut Creek Wastewater Treatment Plant to control filamentous organisms and increase the alkalinity within the aeration basins. The denitrification configuration that the staff implemented was unconventional because no structural changes were made to the aeration basins to encourage denitrification. However, the system functioned well and allowed operators to turn off one of the two air blowers, which saves the plant a significant amount of energy. The current operation has occasional problems, where the alkalinity in the aeration basin decreases or the effluent ammonia increases. When the alkalinity decreases to the point where the pH drops to near 6.0, operators are forced to add chemicals to increase the alkalinity. When the effluent ammonia increases to near the permitted concentration (2.0 mg NH₃-N/L),operators are forced to turn back on the second blower which eliminates the anoxic zone. These problems occur most often during the winter, when the wastewater is the coldest. The wastewater temperature at Walnut Creek varies from a high of 30°C during the summer to a low of 18°C during the winter. The goal of this research was the identification of ways to make the operation more robust which would prevent the need for chemical addition and minimize the use of the second blower. Laboratory-scale reactors were operated to assess possible improvements that could be made to the operation and configuration of the denitrification system at Walnut Creek. The data observed in the laboratory scale experiments showed that the population of denitrifying bacteria limits denitrification and is especially important during the winter. Increasing the solids retention time to 20 days appeared to be the best way to increase the population of denitrifying bacteria and improve denitrification. Improvements can also be made by increasing the volume of the anoxic zone. Increasing the volume of wastewater and biomass recycled will most likely not benefit denitrification until other improvements have been made. Recommendations to the City of Austin Water Utility include the following: 1) increase the solids retention time at Walnut Creek, 2) Increase the volume of the anoxic zone, 3) Separate the anoxic zone from the aerobic section of each aeration basin, 4) During the winter, operate the flow equalization basins to reduce the dissolved oxygen entering the anoxic zone, 5) Continually mix some of the effluent from the aeration basins with the primary effluent in the flow equalization basins. / text

Increasing levels of nitrate (NO3-) in groundwater have become a major environmental and health concern. In situations where NO3-concentrations in the soil-water system pose an environmental hazard, water table management may be a desirable practice to reduce such pollution. Careful management of N applications is also believed to reduce NO3- levels. / A field experiment was conducted in 1996 and 1997 at St. Emmanuel, Quebec, about 30 km South-West of Macdonald Campus of McGill University, to investigate the effect of water table management (WTM) and N fertilizer combinations on potentially leachable NO3- and denitrification rates in the top soil layer (0--0.15 m). The field was planted with monocrop corn (Zea mays. L) in both years. Treatments consisted of a factorial combination of two water table managements, free drainage (FD) and subirrigation (SI) (about 1.0 m and 0.6 m, respectively, below the soil surface) and two N fertilizer rates, 200 kg ha-1 (N200) and 120 kg ha-1 (N120). / Water table management had a significant effect on reducing NO3 - concentrations in the soil profile. Subirrigation treatment reduced NO3- in the top soil layer by 41% and 15% in 1996 and 1997, respectively. Similarly, NO3 - levels were 50% and 20% lower in N120 compared to N200 treatment. / Climatic conditions (rainfall and temperature) played a large role in regulating denitrification rates. Due to drier and cooler conditions in 1997, denitrification rates were lower compared to 1996, leaving more NO3 - in the soil profile. Following harvest, this high NO 3- concentration may be subject to leaching.

The overall goal of this study was to investigate the possibility of reducing NO3-- concentration in the lower soil horizon by promoting denitrification. The study looked at an inexpensive remediation practice for subsurface-drained fields in order to degrade N0j' and consequently diminish NO3-- pollution. The experiments were conducted on a corn field at the Macdonald Campus Farm. The field was composed of sandy loam soil underlined by a clay layer. / In this study, sucrose was injected at a concentration of 20 mg L --1 within the subirrigation water. Furthermore, the water table was maintained at approximately 70 cm from the soil surface in order to create an anaerobic environment that allows for denitrification. The purpose was to furnish dissolved organic carbon (DOC) to the resident microorganisms in order to carryout a higher amount of denitrification. / The NO3-- levels in ground water were monitored weekly. Gas samples were taken from the field to observe whether N2O emissions increased as a result of the treatment This was considered to be important since N2O is a greenhouse gas. / It was found that the addition of sucrose significantly resulted in a faster rate of denitrification. The higher biological activity severely damped the NO3-- peaks in the subsoil, which arose following major rainfall events. / The addition of sucrose at 20 mg L--1 was found to be significant in lowering the amounts of released N2O. Therefore, adding DOC in the subirrigation water not only helped remediate water contamination but also served as a remedy to atmospheric pollution. / In addition, the hydraulic conductivity of the soil was monitored to ensure that no bioclogging arose from an anticipated rise in the bacterial population due to the addition of sucrose. It was found that the addition of sucrose at 20 mg L--1 did not contribute in anyway to reduce the soil's hydraulic conductivity.

The fate of fertilizer N is of primary concern for both agricultural productivity and environmental quality. Concerns include denitrification, leaching losses, mineralization of organic N as plant available N. Denitrification is an important source of N$ sb2$O, a greenhouse gas but field measurements are difficult. Two methods of measuring denitrification are soil core (SC) incubation and closed chamber (CC) methods. These methods were assessed on soil under monoculture corn, monoculture soybean, and alfalfa in a corn soybean alfalfa rotation. Greater concentrations were found in the CC method than the SC method. Denitrification rates ranged from less than 15 g N ha$ sp{-1}$h$ sp{-1}$ to nearly 2000 g N ha$ sp{-1}$h$ sp{-1}$. The CC method was more sensitive to treatment effects. The denitrification rates were dependent on the soil type, being higher on soils with high clay content. The variables that had the highest degree of relationship with denitrification were water filled pore space, soil NH$ sb4$-N and NO$ sb3 $,-N concentrations. Higher rates of N increased denitrification. As to assessment of available soil N, this was accomplished N and C mineralization measurements. Potentially mineralizable N(N$ sb0$) ranged from 144 mg N kg$ sp{-1}$ to 30.3 mg N kg$ sp{-1}$. Higher rates of organic amendment resulted in higher measured values on Brandon soil while higher rates of inorganic N on Chicot and Ste. Rosalie soils caused no change in mineralizable N or respired C. Total N, organic C, water soluble organic C (WSOC) and microbial biomass C (MBC) increased with increasing amounts of organic or inorganic N amendment on Brandon soil. Higher rates of inorganic N resulted in lower WSOC and MBC on Chicot and Ste. Rosalie soils. Nitrogen mineralized, C respired, total N, organic C, WSOC and MBC were all related to soil texture. MBC and WSOC were found to have a strong positive relationship with potentially mineralizable N.

The use of agricultural chemicals, such as nitrogen fertilizers in corn production, often results in water pollution. This research, comprising three parts, was designed to investigate the effects of nitrogen fertilizer application rates, water table management, and corn cropping systems on drainage water quality. The first part was a field study, to investigate the impact of two cropping systems and water table management on nitrate loss through tile drainage. The considered water table treatments were free drainage, and subirrigation with target water table depths at 0.5 m or 0.75 M below the soil surface. Corn (Zea mays L.) monoculture and corn intercropped with annual ryegrass (Lolium multiflorum Lam.) were investigated. The highest annual tile drainage losses of 21.9 kg N/ha were measured in monocropped, freely draining plots. Subirrigation with a water table depth of 0.5 m reduced tile drainage loss of N by over 70%, and intercropping corn with ryegrass under free drainage reduced leaching losses by 50%. / The second part of the research was a simulation study with the water quality model, DRAINMOD-N. The water quality impact of fertilizer application rate under free drainage, subirrigation and controlled drainage was evaluated. Leaching losses, denitrification and N accumulation in the soil profile were investigated. Using data obtained from the field experiment, the performance of DRAINMOD-N was evaluated. DRAINMOD-N assumes that denitrification follows first order kinetics, contrary to field measurements which showed little correlation between denitrification rate and NO$ sb3 sp-$-N concentration. Therefore, DRAINMOD-N was modified by replacing the original denitrification function with the Michaelis-Menten relationship. In so doing, denitrification is expressed as a first order process when NO$ sb3 sp-$-N concentration limits denitrification, and as a zero order process for non-limiting NO$ sb3 sp-$-N concentration. / For denitrification to be a decision making criterion of water table management, inexpensive but reliable measurement techniques are required. Thus, the purpose of the final part of this research was to formulate a technique for measuring real-time denitrification rate. Denitrification rate could be expressed as a function of soil redox potential (Eh) and temperature. Laboratory and field studies showed that factors such as soil nitrate and organic carbon had negligible effect on denitrification rate. Therefore, it can be concluded that for most agricultural soil, Eh and soil temperature will satisfactorily describe denitrification variation.

Thesis (M.Tech.: Biotechnology)-Dept. of Biotechnogy, Durban Institute of Technology, 2005 xvi, 85 leaves : ill. ; 31 cm / Atmospheric nitrogen pollution is on the increase and human activities are directly or indirectly responsible for the generation of the various nitrogen polluting compounds. This can lead to the two major problems of eutrophication and groundwater pollution. Therefore, the removal of nutrients such as nitrogen and phosphorus from wastewater is important. Nitrogen removal from wastewater is achieved by a combination of nitrification and denitrification. Thus, there is a need to identify and characterise heterotrophic denitrifying bacteria involved in denitrification in wastewater treatment systems. The aim of this study, therefore, was to characterise heterotrophic denitrifying bacteria through detailed biochemical and molecular analysis, to facilitate the understanding of their functional role in wastewater treatment systems. Drysdale (2001) isolated heterotrophic denitrifiers to obtain a culture collection of 179 isolates. This culture collection was used to screen for nitrate and nitrite reduction using the colorimetric biochemical nitrate reduction test. The isolates were thereafter Gram stained to assess their gram reaction, cellular and colonial morphology. Based on these results identical isolates were discarded and a culture collection of approximately 129 isolates remained. The genetic diversity of the culture collection was investigated by the analysis of polymerase chain reaction (PCR)-amplified 16S ribosomal DNA (rDNA) fragments on polyacrylamide gels using denaturing gradient gel electrophoresis (DGGE). Thus DNA fragments of the same length but different nucleotide sequences were effectively separated and microbial community profiles of eight predominant isolates were created. Batch experiments were conducted on these eight isolates, the results of which ultimately confirmed their characterisation and placed them into their four functional groups i.e. 3 isolates were incomplete denitrifiers, 2 isolates were true denitrifiers, 2 isolates were sequential denitrifiers and 1 isolate was an exclusive nitrite reducer.

Waste is commonly disposed in landfills, this result in the formation of leachate which needs to be treated to acceptable standards before being discharged to the environment. High concentrations of pollutants, particularly ammonia, in the landfill leachate are persistent even after the closure of the landfill and it requires ad hoc treatment. Treated leachate can still be characterized by high concentrations of nitrates, which exceeds the discharge standards. This phenomenon is observed in the Mariannhill landfill site in Durban, where leachate is nitrified in a Sequencing batch reactor and produces effluent with over 1000 mg/l (Trois et al, 2010a). Denitrification can be used to remove nitrate concentrations, this process occurs under anoxic conditions in the presence of an external carbon source. Denitrification treatment methods utilize chemicals such as methanol and ethanol as carbon sources, but the large scale application of these chemicals is often uneconomical. This research aims at identifying the cost effective treatment system for bio-denitrification that utilizes commercial garden refuse (CGR raw and lightly composted for 10 weeks “CGR 10”) as carbon sources. The feasibility checks for applying these substrates were based on the efficiency and kinetics of nitrate removal over a short and long-term period, thus providing the estimates for operational procedures. Initial characterization tests, batch and column tests were performed in the lab towards achieving the aim of this research. All batch tests achieved 100% of nitrate removal, but CGR raw was faster than CGR 10 with a time difference of 16% and 20% for batches at 100 and 500 mg/L, respectively. The significant difference in the kinetic removal efficiency was observed in batch tests at 2000 mg/L, where CGR raw was about 18 times faster than CGR 10 and about 2 times faster than that of CGR raw at 500 mg/L. Thus, the kinetics of nitrate removal in CGR raw at 2000 mg/L was suspected to be due to chemical reaction other than biological reaction. In the second set of batch tests the kinetics of nitrate removal for CGR raw was about 3 times that of CGR 10. The column tests, which were operated as continuous flow reactor did not achieve full denitrification due to high flow rate applied. First set of column tests (columns A) used previously used substrates to treat synthetic nitrate solution (500 and 2000 mg/L). Second set of column tests (columns B) used fresh substrates to treat pre-treated landfill leachate with nitrate concentration of about 2000 mg/L. CGR 10 achieved better removal efficiency than CGR raw when treating synthetic solution. Whereas, CGR raw achieved better nitrate removal when treating pre-treated landfill leachate. Decrease in flow rate improved the removal efficiency of the substrates. Dilution of nitrified leachate to about 500 mg/L could improve the efficiency of the substrates. / Thesis (M.Sc.Eng.)-University of KwaZulu-Natal, Durban, 2011.