Bioreduction Of Nitrate Research Articles

Promoting the transformation of green rust for As immobilization with Acidovorax sp. strain BoFeN1

Microbially mediated Fe(II) oxidation has a great potential for attenuating arsenic (As) mobility in an anoxic groundwaters. Green rust (GR), a common Fe(II)-bearing phase in such environments, could be easily oxidized into Fe (oxyhydr)oxides through microbial activity. This study focused on Acidovorax sp. strain BoFeN1, an anaerobic nitrate-reducing Fe(II)-oxidizing (NRFO) bacterium, to promote the transformation of GR. In biotic GR transformation experiments, magnetite formation occurred at [As]ini = 5 mg/L while lepidocrocite and goethite were formed at [As]ini = 10 mg/L. In the absence of bacterium, the GR persisted throughout the 120-h experiment. Meanwhile, with the addition of strain BoFeN1, the final aqueous As concentration significantly decreased from 0.237 to 0.004 mg/L (C0 = 5 mg/L) and from 1.457 to 0.096 mg/L (C0 = 10 mg/L) at 120 h. It was indicated that strain BoFeN1 played a crucial role in promoting the GR transformation and enhancing As immobilization. Further investigations revealed that the role of strain BoFeN1 extended beyond Fe-oxidation. With nitrite (the intermediate of nitrate bioreduction) as oxidizer, lepidocrocite/goethite were formed in the chemical-oxidation system, excluding magnetite. In the Bio - [As]ini = 5 mg/L, the occurrence of lepidocrocite via the bio-oxidation of Fe(II) in GR at 24 h, along with the metabolism of strain BoFeN1 reducing nitrate accompanied with H+ consumption, it should be reasonably deduced that the alkaline micro-environment of periplasm induced by strain BoFeN1 were vital for the transformation of lepidocrocite to magnetite triggered by trace Fe(II). However, in the Bio - [As]ini = 10 mg/L, more As adsorbed on GR inhibiting the adsorption of bacterium, so the alkaline micro-environment had no obvious effect on such transformation. This study helps to understand the interdependence between GR and anaerobic NRFO bacterium, and provides a new perspective for more effective As remediation strategies in anoxic groundwaters.

Magnetite enhances As immobilization during nitrate reduction and Fe(II) oxidation by Acidovorax sp. strain BoFeN1

Arsenic (As) cycling in groundwater is commonly coupled to the biogeochemical cycling of iron (Fe) and the associated transformation of Fe minerals present. Numerous laboratory studies suggested that Fe minerals can act as nucleation sites for further crystal growth and as catalysts for abiotic Fe(II) oxidation. In view of the widespread existence of magnetite in anoxic environments where As is often dissolved, we firstly exploited magnetite to enhance As immobilization during nitrate-reducing Fe(II) oxidation (NRFO) induced by Acidovorax sp. strain BoFeN1, a mixotrophic nitrate-reducing Fe(II)-oxidizing bacterium that can oxidize Fe(II) through both enzymatic and abiotic pathways. Subsequently, we investigated how magnetite affects NRFO and As immobilization. Results demonstrated a significant increase in As(III) removal efficiency from 75.4 % to 97.2 % with magnetite, attributed to the higher amount of NRFO and As(III) oxidation promoted by magnetite. It was found that magnetite stimulated the production of extracellular polymeric substances (EPS), which could decrease the diffusion of nitrate in the periplasm of bacteria and shield them against encrustation, resulting in a more rapid reduction of nitrate in the system with magnetite than that without magnetite. Meanwhile, Fe(II) was almost completely oxidized in the presence of magnetite during the whole 72 h experiment, while in the absence of magnetite, 47.7 % of Fe(II) remained, indicating that magnetite could obviously accelerate the chemical oxidation of Fe(II) with nitrite (the intermediates of nitrate bioreduction). Furthermore, the formation of labile Fe(III), an intermediate product of electron transfer between Fe(II) and magnetite, was reasonably deduced to be vital for anoxic As(III) oxidation. Additionally, the XPS analysis of the solid phase confirmed the oxidation of 43.8 % of As(III) to As(V). This study helps to understand the biogeochemical cycling of Fe and As in the environment, and provides a cost-effective and environmentally friendly option for in situ remediation of As-contaminated groundwater.

Biodegradable microplastics boost dissimilatory nitrate reduction to ammonium (DNRA) process contributing to ammonium nitrogen retention in farmland soils

Microplastics (MPs) pollution in agriculture ecosystems profoundly affects biogeochemical cycles. However, the impacts of MPs, particularly biodegradable MPs, on dissimilatory nitrate reduction to ammonium (DNRA) process remain unclear. Herein, the alterations in denitrification and DNRA pathways and underlying mechanism by different MPs exposures (including conventional nonbiodegradable MPs (polyethylene (PE) and polypropylene (PP)) and biodegradable MPs (polylactic acid (PLA) and polybutylene adipate terephthalate (PBAT)) were explored via soil microcosm experiments accompanied with 15N isotope tracer technology and 16S rRNA high-throughput sequencing. Compared with control, PE and PP increased the rates of denitrification by 117–126% but significantly decreased DNRA rates by 39.3–63.4%, whereas DNRA activities in PLA and PBAT treatments were remarkably boosted by 163–242% and dominated soil nitrate bioreduction (6.67–8.69 μg N g−1 d −1). This differentiation was attributed to the nutrient imbalance induced by MPs and distinctly selective effects of MPs on functional microbes. Microbial interaction mechanism further suggested that limited bioavailable carbon intensified the competition between heterotrophic denitrifiers (e.g., Bacillus, Sphingomonas and Microvirga) and DNRA bacteria (e.g., Luteitalea and Streptomyces) for electron donor in nonbiodegradable MPs treatments, reducing the abundances of DNRA bacteria. However, the high bioavailable C/NOx– ratio resulting from biodegradable MPs degradation released such competition and more greatly promoted the activities of DNRA bacteria than denitrifiers, facilitating ammonium retention in soils. Collectively, this study highlights the promotion of biodegradable MPs on soil ammonium recycle via DNRA process in farmland ecosystems and can shed some light on current plastic mulch-film management toward a sustainable and cleaner agricultural industry.

Nitrate Bioreduction under Cr(VI) Stress: Crossroads of Denitrification and Dissimilatory Nitrate Reduction to Ammonium.

This study explored the response of NO3--N bioreduction to Cr(VI) stress, including reduction efficiency and the pathways involved (denitrification and dissimilatory nitrate reduction to ammonium (DNRA)). Different response patterns of NO3--N conversion were proposed under Cr(VI) suppress (0, 0.5, 5, 15, 30, 50, and 80 mg/L) by evaluating Cr(VI) dose dependence, toxicity accumulation, bioelectron behavior, and microbial community structure. Cr(VI) concentrations of >30 mg/L rapidly inhibited NO3--N removal and immediately induced DNRA. However, denitrification completely dominated the NO3--N reduction pathway at Cr(VI) concentrations of <15 mg/L. Therefore, 30 and 80 mg/L Cr(VI) (R4 and R6) were selected to explore the selection of the different NO3--N removal pathways. The pathway of NO3--N reduction at 30 mg/L Cr(VI) exhibited continuous adaptation, wherein the coexistence of denitrification (51.7%) and DNRA (13.6%) was achieved by regulating the distribution of denitrifiers (37.6%) and DNRA bacteria (32.8%). Comparatively, DNRA gradually replaced denitrification at 80 mg/L Cr(VI). The intracellular Cr(III) accumulation in R6 was 6.60-fold greater than in R4, causing more severe oxidant injury and cell death. The activated NO3--N reduction pathway depended on the value of nitrite reductase activity/nitrate reductase activity, with 0.84-1.08 associated with DNRA activation and 1.48-1.57 with DNRA predominance. Although Cr(VI) increased microbial community richness and improved community structure stability, the inhibition or death of nitrogen-reducing microorganisms caused by Cr(VI) decreased NO3--N reduction efficiency.

Microbial nitrate reduction in propane- or butane-based membrane biofilm reactors under oxygen-limiting conditions

Nitrate contamination has been commonly detected in water environments and poses serious hazards to human health. Previously methane was proposed as a promising electron donor to remove nitrate from contaminated water. Compared with pure methane, natural gas, which not only contains methane but also other short chain gaseous alkanes (SCGAs), is less expensive and more widely available, representing a more attractive electron source for removing oxidized contaminants. However, it remains unknown if these SCGAs can be utilized as electron donors for nitrate reduction. Here, two lab-scale membrane biofilm reactors (MBfRs) separately supplied with propane and butane were operated under oxygen-limiting conditions to test its feasibility of microbial nitrate reduction. Long-term performance suggested nitrate could be continuously removed at a rate of ∼40–50 mg N/L/d using propane/butane as electron donors. In the absence of propane/butane, nitrate removal rates significantly decreased both in the long-term operation (∼2–10 and ∼4–9 mg N/L/d for propane- and butane-based MBfRs, respectively) and batch tests, indicating nitrate bio-reduction was driven by propane/butane. The consumption rates of nitrate and propane/butane dramatically decreased under anaerobic conditions, but recovered after resupplying limited oxygen, suggesting oxygen was an essential triggering factor for propane/butane-based nitrate reduction. High-throughput sequencing targeting 16S rRNA, bmoX and narG genes indicated Mycobacterium/Rhodococcus/Thauera were the potential microorganisms oxidizing propane/butane, while various denitrifiers (e.g. Dechloromonas, Denitratisoma, Zoogloea, Acidovorax, Variovorax, Pseudogulbenkiania and Rhodanobacter) might perform nitrate reduction in the biofilms. Our findings provide evidence to link SCGA oxidation with nitrate reduction under oxygen-limiting conditions and may ultimately facilitate the design of cost-effective techniques for ex-situ groundwater remediation using natural gas.

Insight into the enhancing mechanism of silica nanoparticles on denitrification: Effect on electron transfer and microbial metabolism

Although silica nanoparticles (SiNPs) are produced in large numbers for industrial manufacturing and engineering applications, the effect of SiNPs on biotransformation in the environment is still not clear. In the current study, the effect of SiNPs in enhancing denitrification was investigated, and its mechanism was explored from the perspectives of electron transfer, microbial metabolism and bacterial community structure for the first time. Batch experiments showed that a concentration of SiNPs ranging from 0.05 to 5 g/L enhanced the bioreduction of nitrate. The mechanism study showed that SiNPs accelerated the extracellular electron transfer in the denitrification process due to their electron donating capacity, bonding action, and the secretion of more electron shuttles. During the denitrification process, SiNPs promoted metabolic activity, which mainly consists of promoting enzyme activities and electron transport system activity; these metabolic activity assays were positively correlated with SiNPs according to the structural equation modeling analysis. Moreover, SiNPs affected the composition of the microbial community, including denitrifying functional bacteria, silicon-activating bacteria and electron transfer active bacteria exhibiting a synergistic symbiosis. In addition, it was shown, by investigating two functional group-modified SiNPs, that the carboxyl modified SiNPs had the potential to be applied in nitrogen removal due to their performance and non-toxicity. This study presented a better insight into the role of SiNPs in biological transformation.

Simultaneous bio-reduction of nitrate and Cr(VI) by mechanical milling activated corn straw

Abundant lignocellulose waste is an ideal energy source for environmental bioremediation, but its recalcitrance to bioavailability makes this a challenging prospect. We hypothesized that the disruption of straw’s recalcitrant structure by mechanochemical ball milling would enhance its availability for the simultaneous bioreduction of nitrate and Cr(VI). The results showed that the ball-milling process increased the quantity of water-soluble organic matter released from corn straw and changed the composition of organic matter by strongly disrupting its lignocellulose structure. The increase in ball-milling time increased the specific surface area of the straw and favored the adhesion of microorganisms on the straw surface, which enhanced the bioavailability of the energy in the straw. Substantially increased removal of NO3--N (206.47 ± 0.67 mg/g) and Cr(VI) (37.62 ± 0.09 mg/g) was achieved by using straw that was ball milled for 240 min, which validated that ball milling can improve the utilization efficiency of straw by microorganisms. Cellular and molecular biological analyses showed that ball-milled straw increased microbial energy metabolism and cellular activity related to the electron transport chain. This work offers a potential way to achieve the win-win goal of utilizing agricultural wastes and remediating environmental pollution.

A feasibility study of metal sulfide (FeS and MnS) on simultaneous denitrification and chromate reduction

Metal sulfide-based biological process is considered as a promising biotechnology for next-generation wastewater treatment. However, it is not clear if simultaneous bio-reduction of nitrate and chromate was achievable in this process. This study aimed to evaluate the feasibility of metal sulfides (FeS and MnS) on simultaneous denitrification and chromate reduction in autotrophic denitrifying column bioreactors. Results showed that simultaneous reduction of nitrate and chromate was achieved using metal sulfides (FeS and MnS) as electron donors, in which sulfate was the sole soluble end-product. Apart from the sulfur element in the metal sulfides, Fe(II) and Mn(II) were also involved in nitrate and chromate reduction as indicative by the formation of their oxidative states compounds. In microbial communities, SHD-231 and Thiobacillus were the most predominant bacteria, which might have played important roles in simultaneous denitrification and chromate reduction. Compared to FeS, MnS showed a higher performance on nitrate and chromate removal, which could also reduce the toxic inhibition of chromate on nitrate reduction. According to results of XRD and XPS, as well as a lower sulfate production in the FeS system, FeS might have been covered easily to hydroxides due to its bio-oxidation, which limited mass transfer efficiency and bio-availability of FeS. The findings in this study offered insights in the development of promising approaches for the treatment of toxic and hazardous compounds using metal sulfide.

Iron oxide minerals promote simultaneous bio-reduction of Cr(VI) and nitrate: Implications for understanding natural attenuation

Nitrate and Cr(VI) coexist in aquifers, posing a potential threat to ecological environment and public health. Iron oxide minerals (hematite and magnetite) exist ubiquitously in groundwater, which are hot spots for biogeochemical transformation. However, there is still a knowledge gap anout the effect of iron oxide minerals on bioreduction of nitrate and Cr(VI). Here we observed that iron oxide minerals can significantly improve the ability of microorganisms to simultaneously reduce nitrate and Cr(VI), the reduction rates of nitrate and Cr(VI) increased by 7.3 and 8.5 times, respectively. The addition of minerals reinforced biofilm formation and shaped microbial communities with a new dominant strain of Azoarcus. The expression levels of functional genes were also upregulated, including napA, narG, nfsA, yieF, POD, and CAT. Furthermore, nitrate and chromate reductases' activities increased by 11 and 5 folds, respectively. These results demonstrated that iron oxide minerals participated in the bio-transformation of nitrate and Cr(VI) co-contamination, alleviating oxidative stress, shaping the microbial community, and ultimately accelerating bio-transformation. These findings offer a window into the biological transformation of co-contamination in the presence of iron oxide minerals, and insights to reveal strategies for microbial detoxification and to develop promising approaches for dealing with complex pollution conditions.

Nitrate bioreduction dynamics in hyporheic zone sediments under cyclic changes of chemical compositions

Hyporheic zone (HZ) is the hotspot of contaminant biotransformation in ecosystems that affects water quality in both groundwater and river water. HZ is characterized by its dynamic changes in water flow direction that lead to the cyclic changes in aqueous chemical compositions such as dissolved oxygen (DO), nitrate, dissolved organic carbon, and their concentrations. Here we report a study on the effects of cyclic changes in the chemical conditions on the rates of nitrate bioreduction in sediments from a hyporheic zone, and the corresponding dynamics of the synthesis and decay of nitrate reductases. The results indicated that the rate of nitrate bioreduction in the second and third cycles was faster than that in the first one because the nitrate reductases produced in the first cycle still partially maintained their reactivity. The presence of DO significantly decreased the rates of nitrate reduction and nitrate reductase synthesis, but such an inhibition effect diminished in the later cycle. The coupled changes in nitrate concentrations and functional enzymes and genes abundances were generally consistent with an enzyme-based biogeochemical kinetic model. Overall the results indicated that enzyme growth dynamics and functions were complexly affected by the cyclic changes of environmental conditions, and implied that HZ sediments have a memory-type system behavior in reducing nitrate that would be affected by the frequency and magnitude of groundwater and river water exchanges, and chemical compositions in the groundwater and river water.

Bioreduction of nitrate in high-sulfate water using a hydrogen-based membrane biofilm reactor equipped with a separate carbon dioxide module

A hydrogen-based membrane biofilm reactor (MBfR) equipped with separate hydrogen (H2) and carbon dioxide (CO2) adjustable gas-transfer modules was used to explore denitrification in a water containing a high concentration of sulfate (SO42−) (500 mg S/L). CO2 was used for pH control and as the sole carbon source for long-term operation of the MBfR, during which >90% denitrification was achieved. Short-term experiments demonstrated that increasing the SO42− loading had negligible effect on nitrate (NO3−) reduction, but SO42− reduction was strongly suppressed by high NO3− loading regardless of H2 availability. Sulfide oxidation accompanied with SO42− reduction occurred in the biofilm, and the intermediate element sulfur was further oxidized to SO42−. Results of high-throughput sequencing suggest that sulfide oxidation was carried out by the known sulfide-oxidizing bacteria: Starkeya and Xanthobacter. And sulfide oxidation coupled to denitrification was a primary reason for minimal net SO42− reduction when the NO3− loading was high enough. This study demonstrates the promise of effective denitrification without net SO42− reduction in MBfR systems and documents the important sulfur-conversion processes.

Motility of Shewanella oneidensis MR-1 Allows for Nitrate Reduction in the Toxic Region of a Ciprofloxacin Concentration Gradient in a Microfluidic Reactor.

Subsurface environments often contain mixtures of contaminants in which the microbial degradation of one pollutant may be inhibited by the toxicity of another. Agricultural settings exemplify these complex environments, where antimicrobial leachates may inhibit nitrate bioreduction, and are the motivation to address this fundamental ecological response. In this study, a microfluidic reactor was fabricated to create diffusion-controlled concentration gradients of nitrate and ciprofloxacin under anoxic conditions in order to evaluate the ability of Shewanella oneidenisis MR-1 to reduce the former in the presence of the latter. Results show a surprising ecological response, where swimming motility allow S. oneidensis MR-1 to accumulate and maintain metabolic activity for nitrate reduction in regions with toxic ciprofloxacin concentrations (i.e., 50× minimum inhibitory concentration, MIC), despite the lack of observed antibiotic resistance. Controls with limited nutrient flux and a nonmotile mutant (Δ flag) show that cells cannot colonize antibiotic rich microenvironments, and this results in minimal metabolic activity for nitrate reduction. These results demonstrate that under anoxic, nitrate-reducing conditions, motility can control microbial habitability and metabolic activity in spatially heterogeneous toxic environments.

Bromate and Nitrate Bioreduction Coupled with Poly-β-hydroxybutyrate Production in a Methane-Based Membrane Biofilm Reactor.

This work demonstrates bromate (BrO3-) reduction in a methane (CH4)-based membrane biofilm reactor (MBfR), and it documents contrasting impacts of nitrate (NO3-) on BrO3- reduction, as well as formation of poly-β-hydroxybutyrate (PHB), an internal C- and electron-storage material. When the electron donor, CH4, was in ample supply, NO3- enhanced BrO3- reduction by stimulating the growth of denitrifying bacteria ( Meiothermus, Comamonadaceae, and Anaerolineaceae) able to reduce BrO3- and NO3- simultaneously. This was supported by increases in denitrifying enzymes (e.g., nitrate reductase, nitrite reductase, nitrous-oxide reductase, and nitric-oxide reductase) through quantitative polymerase chain reaction (qPCR) analysis and metagenomic prediction of these functional genes. When the electron donor was in limited supply, NO3- was the preferred electron acceptor over BrO3- due to competition for the common electron donor; this was supported by the significant oxidation of stored PHB when NO3- was high enough to cause electron-donor limitation. Methanotrophs (e.g., Methylocystis, Methylomonas, and genera within Comamonadaceae) were implicated as the main PHB producers in the biofilms, and their ability to oxidize PHB mitigated the impacts of competition for CH4.

Concurrent microbial reduction of high concentrations of nitrate and perchlorate in an ion exchange membrane bioreactor.

We investigated effective simultaneous removal of high loads of nitrate and perchlorate from synthetic groundwater using an ion exchange membrane bioreactor (IEMB). The aim of this research was to characterize both transport aspects and biodegradation mechanisms involved in the treatment process of high loads of the two anions. Biodegradation process was proven to be efficient with over 99% efficiency of both perchlorate and nitrate, regardless of their load. The maximum biodegradation rates were 18.3 (mmol m(-2) h(-1) ) and 5.5 (mmol m(-2) h(-1) ) for nitrate and perchlorate, respectively. The presence of a biofilm on the bio-side of the membrane only slightly increased the nitrate and perchlorate transmembrane flux as compared to the measured flux during a Donnan dialysis experiment where there is no biodegradation of perchlorate and nitrate in the bio-compartment. The nitrate flux in presence of a biofilm was 18.3 (±1.9) (mmole m(-2) h(-1) ), while without the biofilm, the flux was 16.9 (±1.5) (mmole m(-2) h(-1) ) for the same feed inlet nitrate concentration of 4 mM. The perchlorate transmembrane flux increased similarly by an average of 5%. Samples of membrane biofilm and suspended bacteria from the bio-reactor were analyzed for diversity and abundance of the perchlorate and nitrate reducing bacteria. Klebsiella oxytoca, known as a glycerol fermenter, accounted for 70% of the suspended bacteria. In contrast, perchlorate and nitrate reducing bacteria predominated in the biofilm present on the membrane. These results are consistent with our proposed two stage biodegradation mechanism where glycerol is first fermented in the suspended phase of the bio-reactor and the fermentation products drive perchlorate and nitrate bio-reduction in the biofilm attached to the membrane. These results suggest that the niche exclusion of microbial populations in between the reactor and membrane is controlled by the fluxes of the electron donors and acceptors. Such a mechanism has important implications for controlling the bio-reduction reaction in the IEMB when using glycerol as a carbon source and allowing treating a complex contamination of high concentrations of perchlorate and nitrating in groundwater and successfully biodegrading them to non-hazardous components. Biotechnol. Bioeng. 2016;113: 1881-1891. © 2016 Wiley Periodicals, Inc.

Nitrate bioreduction in redox-variable low permeability sediments

Low permeability zone (LPZ) can play an important role as a sink or secondary source in contaminant transport in groundwater system. This study investigated the rate and end product of nitrate bioreduction in LPZ sediments. The sediments were from the U.S. Department of Energy's Hanford Site, where nitrate is a groundwater contaminant as a by-product of radionuclide waste discharges. The LPZ at the Hanford site consists of two layers with an oxidized layer on top and reduced layer below. The oxidized layer is directly in contact with the overlying contaminated aquifer, while the reduced layer is in contact with an uncontaminated aquifer below. The experimental results showed that nitrate bioreduction rate and end-product differed significantly in the sediments. The bioreduction rate in the oxidized sediment was significantly faster than that in the reduced one. A significant amount of N2O was accumulated in the reduced sediment; while in the oxidized sediment, N2O was further reduced to N2. RT-PCR analysis revealed that nosZ, the gene that codes for N2O reductase, was below detection limit in the reduced sediment. Batch experiments and kinetic modeling were performed to provide insights into the role of organic carbon bioavailability, biomass growth, and competition between nitrate and its reducing products for electrons from electron donors. The results revealed that it is important to consider sediment redox conditions and functional genes in understanding and modeling nitrate bioreduction in subsurface sediments. The results also implied that LPZ sediments can be important sink of nitrate and a potential secondary source of N2O as a nitrate bioreduction product in groundwater.

Bioreduction of nitrate in a hydrogen-based membrane biofilm reactor using CO2 for pH control and as carbon source

The hydrogen-based membrane biofilm reactor (MBfR) is efficient in removing nitrate in oligotrophic water, but pH increase due to denitrification is a tough problem in practice. To control pH, we investigated a novel MBfR with double bundles of hollow fibers (one for H2 supply and the other for CO2 supply). CO2 was used for pH control and as the carbon source. The Fick’s first law and steady state permeation tests were adopted to determine the CO2 permeability of polyvinyl chloride (PVC) hollow fibers used in this study. The CO2 diffusion coefficient in the membrane was 3.8375×10−7m2/d @ 25°C. The CO2 diffusion coefficient was verified by a nitrate bioreduction experiment in the MBfR. The calculated supply of CO2 based on the CO2 diffusion coefficient is consistent with the experimental results: The total nitrogen removal rate was more than 99% and the pH is stable at around 7.45. CO2 diffused through the membrane is sufficient to be the sole carbon source. We confirmed that the novel MBfR with double bundles was stable and efficient. This research provides direct guidance to the design and operation of this new type MBfR.

Simultaneous bioreduction of nitrate and chromate using sulfur-based mixotrophic denitrification process

This study aims at evaluating simultaneous chromate and nitrate reduction using sulfur-based mixotrophic denitrification process in a column reactor packed with elemental sulfur and activated carbon. The reactor was supplemented with methanol at C/N ratio of 1.33 or 2. Almost complete denitrification was achieved at influent NO3−–N and Cr(VI) concentrations of 75mg/L and 10mg/L, respectively, and 3.7h HRT. Maximum denitrification rate was 0.5g NO3−–N/(L.d) when the bioreactor was fed with 75mg/L NO3−–N, 150mg/L methanol and 10mg/L Cr(VI). The share of autotrophic denitrification was between 12% and 50% depending on HRT, C/N ratio and Cr(VI) concentration. Effluent total chromium was below 50μg/L provided that influent Cr(VI) concentration was equal or below 5mg/L. DGGE results showed stable microbial community throughout the operation and the presence of sulfur oxidizing denitrifying bacteria (Thiobacillus denitrificans) and Cr(VI) reducing bacteria (Exiguobacterium spp.) in the column bed.

Strontium sorption and precipitation behaviour during bioreduction in nitrate impacted sediments

The behaviour of strontium (Sr2+) during microbial reduction in nitrate impacted sediments was investigated in sediment microcosm experiments relevant to nuclear sites. Although Sr2+ is not expected to be influenced directly by redox state, bioreduction of nitrate caused reduced Sr2+ solubility due to an increase in pH during bioreduction and denitrification. Sr2+ removal was greatest in systems with the highest initial nitrate loading and consequently more alkaline conditions at the end of denitrification. After denitrification, a limited re-release of Sr2+ back into solution occurred coincident with the onset of metal (Mn(IV) and Fe(III)) reduction which caused minor pH changes in all microcosms with the exception of the bicarbonate buffered system with initial nitrate of 100mM and final pH>9. In this system ~95% of Sr2+ remained associated with the sediment throughout the progression of bioreduction. Analysis of this pH 9 system using X-ray absorption spectroscopy (XAS) and electron microscopy coupled to thermodynamic modelling showed that Sr2+ became partially incorporated within carbonate phases which were formed at higher pH. This is in contrast to all other systems where final pH was <9, here XAS analysis showed that outer sphere Sr2+ sorption predominated. These results provide novel insight into the likely environmental fate of the significant radioactive contaminant, 90Sr, during changes in sediment biogeochemistry induced by bioreduction in nitrate impacted nuclear contaminated environments.

Bioreduction of nitrate in groundwater using a pilot-scale hydrogen-based membrane biofilm reactor

A long-term pilot-scale H2-based membrane biofilm reactor (MBfR) was tested for removal of nitrate from actual groundwater. A key feature of this second-generation pilot MBfR is that it employed lower cost polyester hollow fibers and still achieved high loading rate. The steady-state maximum nitrate surface loading at which the effluent nitrate and nitrite concentrations were below the Maximum Contaminant Level (MCL) was at least 5.9 g·N·(m2·d)−1, which corresponds to a maximum volumetric loading of at least 7.7 kg·N·(m3·d) −1. The steady-state maximum nitrate surface area loading was higher than the highest nitrate surface loading reported in the first-generation MBfRs using composite fibers (2.6 g·N·(m2·d)−1). This work also evaluated the H2-utilization efficiency in MBfR. The measured H2 supply rate was only slightly higher than the stoichiometric H2-utilization rate. Thus, H2 utilization was controlled by diffusion and was close to 100% efficiency, as long as biofilm accumulated on the polyester-fiber surface and the fibers had no leaks.

Bio-reduction of nitrate from groundwater using a hydrogen-based membrane biofilm reactor

A hydrogen-based membrane biofilm reactor (MBfR) using H 2 as electron donor was investigated to remove nitrate from groundwater. When nitrate was first introduced to the MBfR, denitrification took place on the shell side of the membranes immediately, and the effluent concentration of nitrate continuously decreased with 100% removal rate on day 45 under the influent nitrate concentration of 5 mg NO 3 −-N/L, which described the acclimating and enriching process of autohydrogenotrophic denitrification bacteria. A series of short-term experiments were applied to investigate the effects of hydrogen pressures and nitrate loadings on denitrification. The results showed that nitrate reduction rate improved as H 2 pressure increasing, and over 97% of total nitrogen removal rate was achieved when the nitrate loading increased from 0.17 to 0.34 g NO 3 −-N/(m 2·day) without nitrite accumulation. The maximum denitrification rate was 384 g N/(m 3·day). Partial sulfate reduction, which occurred in parallel to nitrate reduction, was inhibited by denitrififcation due to the competition for H 2. This research showed that MBfR is effective for removing nitrate from the contaminated groundwater.