Anoxic Nitrate Research Articles

Anoxic nitrification with carbon-based materials as terminal electron acceptors

This study investigates the potential of humic substances (HS) and graphene oxide (GO), as extracellular electron acceptors (EEA) for nitrification, aiming to explore alternatives to sustain this process in wastewater treatment systems. Experimental results demonstrate the conversion of ammonium to nitrate (up to 87 % of conversion) coupled to the reduction of either HS or GO by anaerobic consortia. Electron balance confirmed the contribution of HS and GO to ammonium oxidation. Tracer analysis in incubations performed with 15NH4+ demonstrated 15NO3- as the main product with a minor fraction ending as 29N2. Phylogenetic analysis identified Firmicutes, Euryarchaeota, and Chloroflexi as the microbial lineages potentially involved in anoxic nitrification linked to HS reduction. This study introduces a new avenue for research in which carbon-based materials with electron-accepting capacity may support the anoxic oxidation of ammonium, for instance in bioelectrochemical systems in which carbon-based anodes could support this novel process.

Denitrification, anammox, and dissimilatory nitrate reduction to ammonium across a mosaic of estuarine benthic habitats

AbstractEstuaries play a key role in moderating the flow of nitrogen (N) to marine ecosystems. However, the magnitude of this N removal can vary dramatically both within and between estuaries due to the benthic habitats present. Here, we compare denitrification, coupled nitrification–denitrification, anammox, and dissimilatory nitrate reduction to ammonium (DNRA) across a mosaic of benthic habitats in the subtropical Noosa River Estuary, Australia. Using 15N tracer techniques and passive pore‐water samplers, we show that coupled nitrification–denitrification was the dominant pathway for N2 production across all habitats, with higher rates in vegetated habitats (10–70 μmol N m−2 h−1) compared to bare sediments (0.9–2 μmol N m−2 h−1). Unusual pore‐water profiles in the macroalgal sediments suggest the presence of sulfur‐driven anoxic nitrification of NH4+ to NO3− and N2. A benthic N budget showed that combined denitrification and coupled nitrification–denitrification accounted approximately 96% of the N2 production, while DNRA accounted for 9% of total NO3− reduction pathways in the Noosa River Estuary. The macroalgae habitat contributed 76% of total N removal via N2 production and 65% of N retention via DNRA, despite accounting for only 25% of the total surface area. We show a strong relationship between seagrass and macroalgae area and N2 production (r2 = 0.8; p < 0.01), and as such, the capacity to mitigate reactive N loads in estuaries may decrease with the large‐scale loss of seagrass and other vegetated habitats.

An isotopic study of abiotic nitrite oxidation by ligand-bound manganese (III)

Redox transformations of nitrogen (N) play a critical role in determining its speciation and biological availability, thus defining the magnitude and extent of productivity in many ecosystems. A range of important nitrogen transformations often co-occur in regions hosting other redox-active elements, including sulfur, iron, and manganese (Mn), especially along sharp redox gradients within aquatic sediments. This proximity in “redox real estate” produces conditions under which multi-element interactions and coupled cycling are thermodynamically favored. While previous work has reported anoxic nitrification linked to the presence of manganese (Mn) oxides in sediments, a clear connection between the cycling of Mn and N has remained elusive. Soluble Mn(III), which is stabilized via ligand-complexation, has recently been shown to represent the dominant dissolved Mn species in many environments. Here, we examined the reactivity of ligand-stabilized Mn(III) with nitrite, using natural abundance stable nitrogen and oxygen isotopes to explore reaction dynamics under a range of conditions. Oxidation of nitrite to nitrate by Mn(III)-pyrophosphate proceeded abiotically under both oxygen replete and nitrogen-purged conditions. Kinetics and isotope systematics of this reaction were measured over a range of pH (5–8), with reaction rates decreasing with increasing pH. Under all treatments, an inverse kinetic isotope effect of −19.9 ± 0.7‰ was observed for N, remarkably similar to previously documented fractionation by nitrite-oxidizing organisms. Experiments using 18O-labeled water confirmed that the source of the additional oxygen atom was from water. These findings suggest that nitrite oxidation in environments hosting abundant ligand-bound Mn(III), including porewaters, estuaries, and coastal waters, may be facilitated in part by abiotic reactions with Mn, even under functionally anoxic conditions.

Denitrification performance and microbial versatility in response to different selection pressures

This study investigated functional dynamics of microbial community in response to different selection pressures, with a focus on denitrification. Suspended-biomass experiments demonstrated limited aerobic and relatively higher anoxic nitrate and nitrite reduction capabilities; the highest NO2-N and NO3-N removal rates were 1.3 ± 0.1 and 0.74 ± 0.01 in aerobic and 1.4 ± 0.05 and 3.4 ± 0.1 mg/L.h in anoxic media, respectively. Key potential denitrifiers were identified as: (i) complete aerobic denitrifiers: Dokdonella, Flavobacterium, and Ca. Accumulibacter; (ii) complete anoxic denitrifiers: Acinetobacter, Pseudomonas, Arcobacter, and Comamonas; (iii) incomplete nitrite denitrifier: Diaphorobacter (aerobic/anoxic), (iv): incomplete nitrate denitrifiers: Thauera (aerobic/anoxic) and Zoogloea (strictly-aerobic). Granular biomass removed 72 mg/L NH4-N with no NOx− accumulation. Heterotrophic nitrification and aerobic denitrification were proposed as the principal nitrogen removal pathway in granular reactors, potentially performed by two key organisms Thuaera and Flavobacterium. Biodiversity analysis suggested that the selection pressure of nourishment condition was the decisive factor for microbial selection and nitrogen removal mechanism.

Influence of application of manganese ore in constructed wetlands on the mechanisms and improvement of nitrogen and phosphorus removal

Vertical up-flow constructed wetlands (CWs) with manganese ore (Mn ore) as media (M-CWs) were developed to treat simulated polluted river water. The results showed that the average removal efficiencies for NH4-N, NO3-N, TN and TP were 91.74%, 83.29%, 87.47% and 65.12% in M-CWs, respectively, which were only 79.12%, 72.90%, 75.85% and 43.23% in the CWs without Mn ore (C-CWs). Nutrient mass balance showed that nitrogen (N) removal was improved by enhanced microbial processes, media storage and plant uptake in M-CWs. Moreover, almost 50% of phosphorus (P) was retained by media storage because of the adsorption processes on Mn ore. It was found that addition of Mn ore enhanced denitrification as the relative abundance of denitrifying bacteria increased. The produced Mn(II) and more abundant Gammaproteobacteria confirmed alternative N removal pathways including anoxic nitrification coupled to Mn ore reduction and denitrification using Mn(II) as electron donor. Mn(II) concentration in the effluent of M-CWs was below the drinking water limit of 0.1 mg/L, which makes them environmentally-friendly.

Microbial electricity driven anoxic ammonium removal

Removal of nitrogen, mainly in form of ammonium (NH4+), in wastewater treatment plants (WWTPs) is a highly energy demanding process, mainly due to aeration. It causes costs of about half a million Euros per year in an average European WWTP. Alternative, more economical technologies for the removal of nitrogen compounds from wastewater are required. This study proves the complete anoxic conversion of ammonium (NH4+) to dinitrogen gas (N2) in continuously operated bioelectrochemical systems at the litre-scale. The removal rate is comparable to conventional WWTPs with 35 ± 10 g N m−3 d−1 with low accumulation of NO2−, NO3−, N2O. In contrast to classical aerobic nitrification, the energy consumption is considerable lower (1.16 ± 0.21 kWh kg−1 N, being more than 35 times less than for the conventional wastewater treatment). Biotic and abiotic control experiments confirmed that the anoxic nitrification was an electrochemical biological process mainly performed by Nitrosomonas with hydroxylamine as the main substrate (mid-point potential, Eox = +0.67 ± 0.08 V vs. SHE). This article proves the technical feasibility and reduction of costs for ammonium removal from wastewater, investigates the underlying mechanisms and discusses future engineering needs.

Effective removal of bromate in nitrate-reducing anoxic zones during managed aquifer recharge for drinking water treatment: Laboratory-scale simulations

The removal of bromate (BrO3−) as a by-product of ozonation in subsequent managed aquifer recharge (MAR) systems, specifically in anoxic nitrate (NO3−)-reducing zones, has so far gained little attention. In this study, batch reactors and columns were used to explore the influence of NO3− and increased assimilable organic carbon (AOC) due to ozonation pre-treatment on BrO3− removal in MAR systems. 8 m column experiments were carried out for 10 months to investigate BrO3− behavior in anoxic NO3−-reducing zones of MAR systems. Anoxic batch experiments showed that an increase of AOC promoted microbial activity and corresponding BrO3− removal. A drastic increase of BrO3− biodegradation was observed in the sudden absence of NO3− in both batch reactors and columns, indicating that BrO3− and NO3− competed for biodegradation by denitrifying bacteria and NO3− was preferred as an electron acceptor under the simultaneous presence of NO3− and BrO3−. However, within 75 days’ absence of NO3− in the anoxic column, BrO3− removal gradually decreased, indicating that the presence of NO3− is a precondition for denitrifying bacteria to reduce BrO3− in NO3−-reducing anoxic zones. In the 8 m anoxic column set-up (retention time 6 days), the BrO3− removal achieved levels as low as 1.3 μg/L, starting at 60 μg/L (98% removal). Taken together, BrO3− removal is likely to occur in vicinity of NO3−-reducing anoxic zones, so MAR systems following ozonation are potentially effective to remove BrO3−.

Low frequency-low voltage alternating electric current-induced anoxic granulation in biofilm-electrode reactor: A study of granule properties

Abstract We characterized anoxic nitrate granules produced by an alternating current biofilm electrode reactor operating at low voltage-low frequency under optimum conditions. Hydrodynamic results revealed that the settling velocity for the granules ranged from 0.12 to 2.85 cm/s, while that of settling velocities ranged from 0.07 to 3.42 cm/s. Granule diameter varied, with a mean mass of 3.65 ± 1.29 mm and corresponding dry mass ranging from 0.52 to 5.64 mg. Roundness ratio of the sampled granules was determined to be 0.78 ± 0.11. Integrity coefficient obtained from a shear strength test was 87.05 ± 2.07% after 2 min and 74.1 ± 4.14% after 5 min. An adhesion test revealed hydrophilic properties of bacteria. The Most probable number (MPN) value was 2.0 × 106 for HDB and 2.0 × 103 for ADB. An apoptosis assay by flow cytometry confirmed that the majority of cells (87.7%) were viable and non-apoptotic (Annexin V-PI) and dehydrogenase activity was 15.05 ± 1.76 μg TF/mg biomass cm−2 d. Comparison of seed and granules by 1H NMR spectra showed different signals in the range of 0.279–1, 1–1.5, and 1.5–7.5 ppm. Therefore, the biofilm in ACBER can be easily granulated and used to generate dense and fast-settling sludge granules.

Microbial mediated anoxic nitrification-denitrification in the presence of nanoscale oxides of manganese

The present study investigates the feasibility of anoxic nitrification-denitrification in the presence of nanoscale oxides of manganese (NSOM). A template-assisted co-precipitation method was employed to synthesize NSOM. XRD and FESEM data reveal that the composite is amorphous in nature, and the particles are in nano regime (15–28 nm). The use of NSOM as an electron acceptor for the microbial mediated anoxic nitrification and the feasibility of re-oxidation of reduced NSOM coupled to denitrification was demonstrated by laboratory scale sequencing batch reactor (SBR). SBR operation lasted for 343 days in 9 different phases by varying the initial NH4+-N (100–500 mg l−1) and cycle time (0.5–2.0 d). Maximum removals of NH4+-N and total nitrogen were 126.2 ± 11.3 mg l−1 d−1 and 109.8 ± 10.9 mg l−1 d−1 respectively, in 0.5 d cycle time. Separate batch experiments were conducted to study the effect of time, pH, temperature, COD/NH4+-N ratio, and inhibitory concentrations of NH4+-N, NO2−-N, and S2−, on the anoxic nitrification process. The developed process is economical as it works in the absence of external supply of aeration and alkalinity, and reduces 70% of the requirement of COD compared to conventional nitrification-denitrification.

Polycyclic Aromatic Hydrocarbon-Induced Changes in Bacterial Community Structure under Anoxic Nitrate Reducing Conditions.

Although bacterial anaerobic degradation of mono-aromatic compounds has been characterized in depth, the degradation of polycyclic aromatic hydrocarbons (PAHs) such as naphthalene has only started to be understood in sulfate reducing bacteria, and little is known about the anaerobic degradation of PAHs in nitrate reducing bacteria. Starting from a series of environments which had suffered different degrees of hydrocarbon pollution, we used most probable number (MPN) enumeration to detect and quantify the presence of bacterial communities able to degrade several PAHs using nitrate as electron acceptor. We detected the presence of a substantial nitrate reducing community able to degrade naphthalene, 2-methylnaphthalene (2MN), and anthracene in some of the sites. With the aim of isolating strains able to degrade PAHs under denitrifying conditions, we set up a series of enrichment cultures with nitrate as terminal electron acceptor and PAHs as the only carbon source and followed the changes in the bacterial communities throughout the process. Results evidenced changes attributable to the imposed nitrate respiration regime, which in several samples were exacerbated in the presence of the PAHs. The presence of naphthalene or 2MN enriched the community in groups of uncultured and poorly characterized organisms, and notably in the Acidobacteria uncultured group iii1-8, which in some cases was only a minor component of the initial samples. Other phylotypes selected by PAHs in these conditions included Bacilli, which were enriched in naphthalene enrichments. Several nitrate reducing strains showing the capacity to grow on PAHs could be isolated on solid media, although the phenotype could not be reproduced in liquid cultures. Analysis of known PAH anaerobic degradation genes in the original samples and enrichment cultures did not reveal the presence of PAH-related nmsA-like sequences but confirmed the presence of bssA-like genes related to anaerobic toluene degradation. Altogether, our results suggest that PAH degradation by nitrate reducing bacteria may require the contribution of different strains, under culture conditions that still need to be defined.

Anoxic nitrate reduction coupled with iron oxidation and attenuation of dissolved arsenic and phosphate in a sand and gravel aquifer

Nitrate has become an increasingly abundant potential electron acceptor for Fe(II) oxidation in groundwater, but this redox couple has not been well characterized within aquifer settings. To investigate this reaction and some of its implications for redox-sensitive groundwater contaminants, we conducted an in situ field study in a wastewater-contaminated aquifer on Cape Cod. Long-term (15year) geochemical monitoring within the contaminant plume indicated interacting zones with variable nitrate-, Fe(II)-, phosphate-, As(V)-, and As(III)-containing groundwater. Nitrate and phosphate were derived predominantly from wastewater disposal, whereas Fe(II), As(III), and As(V) were mobilized from the aquifer sediments. Multiple natural gradient, anoxic tracer tests were conducted in which nitrate and bromide were injected into nitrate-free, Fe(II)-containing groundwater. Prior to injection, aqueous Fe(II) concentrations were approximately 175μM, but sorbed Fe(II) accounted for greater than 90% of the total reactive Fe(II) in the aquifer. Nitrate reduction was stimulated within 1m of transport for 100μM and 1000μM nitrate additions, initially producing stoichiometric quantities of nitrous oxide (>300μMN). In subsequent injections at the same site, nitrate was reduced even more rapidly and produced less nitrous oxide, especially over longer transport distances. Fe(II) and nitrate concentrations decreased together and were accompanied by Fe(III) oxyhydroxide precipitation and decreases in dissolved phosphate, As(III), and As(V) concentrations. Nitrate N and O isotope fractionation effects during nitrate reduction were approximately equal (ε15N/ε18O=1.11) and were similar to those reported for laboratory studies of biological nitrate reduction, including denitrification, but unlike some reported effects on nitrate by denitrification in aquifers. All constituents affected by the in situ tracer experiments returned to pre-injection levels after several weeks. Additionally, Fe(II)-oxidizing, nitrate-reducing microbial enrichment cultures were obtained from aquifer sediments. Growth experiments with the cultures sequentially produced nitrite and nitrous oxide from nitrate while simultaneously oxidizing Fe(II). Field and culture results suggest that nitrogen oxide reduction and Fe(II) oxidation in the aquifer are a complex interaction of coupled biotic and abiotic reactions. Overall, the results of this study demonstrate that anoxic nitrate-dependent iron oxidation can occur in groundwater; that it could control iron speciation; and that the process can impact the mobility of other chemical species (e.g., phosphate and arsenic) not directly involved in the oxidation–reduction reaction.

Using the method of wastewater acidification to improve the efficiency of carbon utilization and nutrient removal in A2N process from a lab-scale operation

Many wastewater treatment processes were developed in order to avoid excessive discharge of nitrogen (N) and phosphorous (P) into the downstream water bodies to mitigate their eutrophication problems. However, one of common problems for these processes is that there is no sufficient carbon source for denitrification. In this research, a continuous anaerobic–anoxic–nitrification process, which was based on reaction of denitrifying phosphorous removing bacteria, was tested to improve carbon utilization and nutrient removal from sewage by employing the method of wastewater acidification. The results have presented that different acidification times could cause remarkable changes in the carbon transformation and utilization. The volatile fatty acids concentration has increased by 56.26±2.46 mg/L during the period of 24-h acidification. Accordingly, the efficiency of carbon source utilization in the anaerobic and anoxic reactors increased by 21.21% and 25.03%, leading to the final removing efficiency of 80.53% total nitrogen and 92.37% total phosphorus, respectively. The effluent water quality could meet the Chinese Discharging Standards for Urban Wastewater Treatment Plants (GB18918–2002) grade A after 8-h acidification. It seemed that acidification had no significant influence on the internal transformation between Glycogen and Poly-β-hydroxybutyrate. Particulate organic carbon was found to be an important potential carbon source, and the removal efficiency of chemical oxygen demand showed no obvious relationship with acidification time as well.

Copper Starvation-inducible Protein for Cytochrome Oxidase Biogenesis in Bradyrhizobium japonicum

Microarray analysis of Bradyrhizobium japonicum grown under copper limitation uncovered five genes named pcuABCDE, which are co-transcribed and co-regulated as an operon. The predicted gene products are periplasmic proteins (PcuA, PcuC, and PcuD), a TonB-dependent outer membrane receptor (PcuB), and a cytoplasmic membrane-integral protein (PcuE). Homologs of PcuC and PcuE had been discovered in other bacteria, namely PCu(A)C and YcnJ, where they play a role in cytochrome oxidase biogenesis and copper transport, respectively. Deletion of the pcuABCDE operon led to a pleiotropic phenotype, including defects in the aa(3)-type cytochrome oxidase, symbiotic nitrogen fixation, and anoxic nitrate respiration. Complementation analyses revealed that, under our assay conditions, the tested functions depended only on the pcuC gene and not on pcuA, pcuB, pcuD, or pcuE. The B. japonicum genome harbors a second pcuC-like gene (blr7088), which, however, did not functionally replace the mutated pcuC. The PcuC protein was overexpressed in Escherichia coli, purified to homogeneity, and shown to bind Cu(I) with high affinity in a 1:1 stoichiometry. The replacement of His(79), Met(90), His(113), and Met(115) by alanine perturbed copper binding. This corroborates the previously purported role of this protein as a periplasmic copper chaperone for the formation of the Cu(A) center on the aa(3)-type cytochrome oxidase. In addition, we provide evidence that PcuC and the copper chaperone ScoI are important for the symbiotically essential, Cu(A)-free cbb(3)-type cytochrome oxidase specifically in endosymbiotic bacteroids of soybean root nodules, which could explain the symbiosis-defective phenotype of the pcuC and scoI mutants.

Resolving the contributions of the membrane-bound and periplasmic nitrate reductase systems to nitric oxide and nitrous oxide production in Salmonella enterica serovar Typhimurium

The production of cytotoxic nitric oxide (NO) and conversion into the neuropharmacological agent and potent greenhouse gas nitrous oxide (N₂O) is linked with anoxic nitrate catabolism by Salmonella enterica serovar Typhimurium. Salmonella can synthesize two types of nitrate reductase: a membrane-bound form (Nar) and a periplasmic form (Nap). Nitrate catabolism was studied under nitrate-rich and nitrate-limited conditions in chemostat cultures following transition from oxic to anoxic conditions. Intracellular NO production was reported qualitatively by assessing transcription of the NO-regulated genes encoding flavohaemoglobin (Hmp), flavorubredoxin (NorV) and hybrid cluster protein (Hcp). A more quantitative analysis of the extent of NO formation was gained by measuring production of N₂O, the end-product of anoxic NO-detoxification. Under nitrate-rich conditions, the nar, nap, hmp, norV and hcp genes were all induced following transition from the oxic to anoxic state, and 20% of nitrate consumed in steady-state was released as N₂O when nitrite had accumulated to millimolar levels. The kinetics of nitrate consumption, nitrite accumulation and N₂O production were similar to those of wild-type in nitrate-sufficient cultures of a nap mutant. In contrast, in a narG mutant, the steady-state rate of N₂O production was ~30-fold lower than that of the wild-type. Under nitrate-limited conditions, nap, but not nar, was up-regulated following transition from oxic to anoxic metabolism and very little N₂O production was observed. Thus a combination of nitrate-sufficiency, nitrite accumulation and an active Nar-type nitrate reductase leads to NO and thence N₂O production, and this can account for up to 20% of the nitrate catabolized.

Nutrient and Trace-Metal Removal by Bauxsol Pellets in Wastewater Treatment

In this study, Bauxsol pellets packed in PVC columns were used to remove nutrients and trace-metals from municipal wastewater during a 6 months field trial. Bauxsol pellet columns showed a high phosphate removal rate via precipitation of PO(4)(3-) with Ca(2+) and Mg(2+) ions: at 90% in the 1st month; at 80% from the second to fifth months; and at 60% in the sixth month. Pellet bound total phosphorus and Colwell phosphorus were 7.3 g/kg and 2 g/kg and are about 20 times the concentrations found in most fertile soils. Trace-metals in effluents were bound, probably irreversibly under the columns' environmental conditions, to the Bauxsol minerals that have high surface area to volume ratios and high charge to mass ratios. Experimental results showed a complex nitrogen cycle operating within the Bauxsol pellet columns including anoxic nitrification, denitrification, and anammox processes. Although a transient pH spike, associated with the release of unreacted CaO from the cement binder used in the pellets, was observed, this may be readily corrected through post-treatment pH adjustment. Hence, the geochemistry of Bauxsol pellets can effectively remove and bind nutrients and trace-metals during wastewater treatment, and further research may show that saturated spent pellets can be used as fertilizer.

Stability and microbial community structure of a partial nitrifying fixed-film bioreactor in long run

A partial nitrification system was investigated for 471 days under DO varying concentrations for assessing its stability and population dynamics. Within 130 days of operation at feed DO concentration of 1.0±0.1 mg/L, more than 85% of nitrite was accumulated. Efficiency deteriorated when the feed DO concentration was increased to 4.2±0.3 mg/L. Nitrite accumulation could not be re-established on decreasing feed DO to 1.0±0.1 mg/L. Even at DO concentration of<0.05 mg/L, nitrate production was observed; a condition termed as anoxic nitrification. NOB was detected in the biomass even under this condition by Fluorescence in-situ hybridization (FISH) analysis. Through 16S rRNA gene sequencing a major fraction of unknown bacterial sequences closely resembling haloalkalophilic bacteria of marine origin were detected. The study indicated that these bacterial species might play a role in anoxic nitrification and that NOB could survive extreme low DO condition.

Microbial Oxidation of Pyrite Coupled to Nitrate Reduction in Anoxic Groundwater Sediment

Although many areas in Denmark are intensively agricultured, the discharge of nitrate from groundwater aquifers to surface water is often lower than expected. In this study it is experimentally demonstrated that anoxic nitrate reduction in sandy sediment containing pyrite is a microbially mediated denitrification process with pyrite as the primary electron donor. The process demonstrates a temperature dependency (Q10) of 1.8 and could be completely inhibited by addition of a bactericide (NaN3). Experimentally determined denitrification rates show that more than 50% of the observed nitrate reduction can be ascribed to pyrite oxidation. The apparent zero-order denitrification rate in anoxic pyrite containing sediment at groundwater temperature has been determined to be 2-3 micromol NO3- kg(-1) day(-1). The in situ groundwater chemistry at the boundary between the redoxcline and the anoxic zone reveals that between 65 and 80% of nitrate reduction in the lower part of the redoxcline is due to anoxic oxidation of pyrite by nitrate with resulting release of sulfate. It is concluded that microbes can control groundwater nitrate concentrations by denitrification using primarily pyrite as electron donor at the oxic-anoxic boundary in sandy aquifers thus determining the position and downward progression of the redox boundary between nitrate-containing and nitrate-free groundwater.

Synthetic dairy wash-water treatment by using an alternating pumped sequencing batch biofilm reactor

The performance of an alternating pumped sequencing batch biofilm reactor process in synthetic diary wash-water treatment was studied. The system comprises two reactors with two identical plastic biofilm modules. Two centrifugal pumps, one connected to each reactor, alternately move the water from one reactor to the other one, resulting in aeration. At three loading rates (336, 501 and 1080 g COD/(m³ d)), total COD and total nitrogen were removed by 91-94% and 46-80%, respectively. Nitrogen was removed from wastewater by denitrification in the anoxic phase and Simultaneous Nitrification and Denitrification (SND) in the aerobic phase.

Anoxic nitrification: Evidence from Humber Estuary sediments (UK)

Conventional understanding of the nitrogen cycle in marine sediments has changed in recent years with the discovery of an alternative pathway for ammonia oxidation via the reduction of manganese oxides (during anoxic nitrification). In anoxic sediments, the potential for manganese oxides to serve as oxidant for nitrification may be considerable yet previous work on manganese-rich sediments has suggested anoxic nitrification may not be significant. In this study, the potential for anoxic nitrification in a range of sediment types was investigated. Laboratory incubation of sediment from three sites on the Humber Estuary, a microbially diverse environment, showed anoxic accumulation of nitrate, nitrite and dinitrogen gas, with and without the addition of synthetic manganese oxides. Incubation experiments confirmed anoxic nitrification as microbially mediated, with heat-killed controls yielding negative results. The anoxic nitrification reaction significantly depleted ammonia concentrations, and occurred simultaneously with manganese-, iron- and sulphate reduction, and methanogenesis. Taken in conjunction with other studies, results suggest anoxic nitrification may not only be dependent on total manganese concentrations but on manganese dynamics. Anoxic nitrification may be explained as a non-steady state reaction, dependent on the recent stability of a sediment system. Physical perturbation of sediments may cause the redistribution and/or introduction of manganese oxides and promote anoxic nitrification. The significance and persistence of anoxic nitrification is likely to depend on the frequency and magnitude of sediment perturbation, which explains why the reaction varies so widely across studied sites, and why it may not occur in some manganese-rich sediment.

The biogeochemistry of a manganese-rich Scottish sea loch: Implications for the study of anoxic nitrification

The anoxic oxidation of ammonia by manganese oxides is a newly recognised pathway for the production of N 2 in sedimentary environments, potentially contributing a significant loss of nitrogen from the world's oceans. Due to the complex recycling of redox species in marine sediments this process is difficult to discern in the natural environment, and is consequently poorly understood. The potential for anoxic nitrification coupled to manganese reduction was investigated through field research and laboratory incubation experiments. Field data from Loch Fyne, a manganese-rich site, did not provide conclusive evidence for anoxic nitrification, although minor accumulation of nitrate was observed in anoxic pore-waters. Incubation of Loch Fyne sediments showed anoxic nitrification to occur, with accumulation of both nitrate and nitrite coincident with removal of ammonia under anoxia, although these observations were not reproduced in repeat experiments. The laboratory evidence for anoxic nitrification confirms the reaction is possible in marine sediments; however, the wider significance of anoxic nitrification remains uncertain. Contrary to previous assumptions about anoxic nitrification, results suggest the reaction may not be dependent on total manganese concentrations and may be inhibited by conventional heterotrophic manganese reduction in manganese-rich sediments.