Hydrogenotrophic Denitrification Research Articles

Controlling carbon dioxide-to-hydrogen ratio to improve hydrogen utilization and denitrification rates of hydrogenotrophic autotrophic denitrification through homoacetogenesis-heterotrophic denitrification pathway

Hydrogenotrophic denitrification, an environment-friendly process for organic-free influents, is limited due to poor hydrogen mass transfer efficiency and significant pH fluctuations. In this study, we manipulated the carbon dioxide-to-hydrogen ratio to improve hydrogenotrophic denitrification. When carbon dioxide-to-hydrogen ratio was 1:1 (carbon dioxide, 200 ml: hydrogen, 200 ml), the hydrogen utilization and denitrification rates were 2.4 times and 3.0 times that when carbon dioxide-to-hydrogen ratio was 0:1 (carbon dioxide, 0 ml: hydrogen, 200 ml), respectively. The pH fluctuation decreased from 3.1±0.3 to 0.2±0.1. Furthermore, the hydrogenotrophic denitrification, acetoclastic denitrification, homoacetogenic, and electron transfer activities of the sludge were improved. A high carbon dioxide-to-hydrogen ratio augmented the acid-producing and heterotrophic denitrifying microorganism populations. By maintaining a high carbon dioxide-to-hydrogen ratio, the dominant hydrogenotrophic autotrophic denitrification pathway was transformed into a homoacetogenesis-heterotrophic denitrification pathway, thereby achieving higher hydrogen utilization and denitrification rates.

Hydrogenotrophic denitrification for treatment of nitrate-contaminated groundwater: Using a microbubble diffuser to optimize performance

Nitrate nitrogen (NO3−-N) contamination of groundwater as a drinking water resource has been detected in many countries and poses a serious threat to human health. This study optimized the performance of a hydrogenotrophic denitrification (HD) reactor for nitrate-contaminated groundwater treatment by introducing a microbubble diffuser (MD), and evaluated its performance stability. The HD reactor continuously maintained effluent NO3−-N and NO2−-N concentrations below the drinking water standard for 200 days under optimized conditions (8 h hydraulic retention time, 10 mL min−1 H2 flow rate, and 50 mg L−1 NO3−-N). The nitrogen removal rate and nitrogen removal efficiency were 142.0 gN m−3 d−1 and 98 %, respectively. Additionally, the HD reactor required 0.02 m3 of H2 for 1 g of nitrogen removal, which indicated that approximately 24 % of the supplied H2 was utilized for HD. The MD produced finer gas bubbles and showed approximately 2.3-times higher gas-dissolving efficiency than an air stone diffuser producing ordinary bubbles. The efficiency is attributed to reactor performance optimization. In addition, Azoarcus spp. was the predominant genus in the HD reactor and accounted for 42.3 % of the total bacteria, suggesting that Azoarcus spp. might be related to HD in the reactor. Functional genes for denitrification (narG and nirS) were continuously detected. The experimental results lead to the development of an effective HD reactor for nitrate-contaminated groundwater treatment—in terms of elimination of H2 supply for denitrification under safe operation—and can facilitate the improvement of water security.

Liquid-gas phase transition enables microbial electrolysis cell to treat organic pollution and synchronously remediate nitrate-contaminated groundwater via hydrogenotrophic denitrification

Groundwater denitrification is challenged by a lack of electron donors and usually requires additional energy input or chemical agents. MEC can convert organic pollution into clean electron donor (hydrogen) through a gas-liquid phase transition. In this study, the MEC was combined with permeable bio-reactive barrier (PRB) as a hybrid system to achieved simultaneous organic waste removal, hydrogen production and groundwater remadition via long-distance hydrogenotrophic denitrification by biogas. The semi-hydrophobic PTFE-coated granular activated carbons (GAC) and hydrophilic GAC were filled as two kinds of biocarriers in PRBs, in which the hydrophobic PTFE surface coating improved hydrogen utilization. The PTFE-coated GAC maked the denitrification performance of PRB insensitive to hydrogen partial pressure within the tested range of 0.01–0.04 MPa and achieved effluent nitrate concentration of <20 mg L−1 within 12 h (The Water Quality Standard for Drinking Water Sources in China). The gas production in MECs was actively pumped to PRBs and spontaneously achieve negative pressure on the MEC side and positive pressure on the PRB side. The actively pumping of biogas induced negative pressure in MEC and positive pressure in PRB. The negative pressure improved current density by 18.6 ± 0.7% in MECs and the MEC-PRB hybrid system achieved a nitrate removal of 85.0 ± 0.8% in actual groundwater with an effluent concentration of 15.0 ± 0.8 mg L−1 at 72 h. This work demonstrated the feasibility of matching MEC and PRB as a novel hybrid system for organic pollution degradation and effective nitrogen removal in electron-donor-lacking groundwater.

Liquid-gas phase transition enables microbial electrolysis and H2-based membrane biofilm hybrid system to degrade organic pollution and achieve effective hydrogenotrophic denitrification of groundwater

Electron-donor Lacking was the limiting factor for the denitrification of oligotrophic groundwater and hydrogenotrophic denitrification provided an efficient approach without secondary pollution. In this study, a hybrid system with microbial electrolysis cell (MEC) assisted hydrogen-based membrane biofilm reactor (MBfR) was established for advanced groundwater denitrification. The liquid-gas phase transition prevented the potential pollution from organic wastes in MEC to groundwater, while the bubble-free diffusion of MBfR promoted hydrogen utilization efficiency. The negative-pressure extraction from MEC and the positive pressure for gas supply into MBfR increased the hydrogen proportion and current density of MEC, and improved the kinetic constant K of the denitrification reaction in MBfR. With actual groundwater, the MEC-MBfR hybrid system achieved a nitrate reduction of 97.8% with an effluent NO3−-N of 2.2 ± 1.0 mg L−1. The hydrogenotrophic denitrifiers of Thauera, Pannonibacter, and Azonexus, dominated the denitrification biofilm on the membrane and elastic filler in MBfR.

Groundwater denitrification using a continuous flow mode hybrid system combining a hydrogenotrophic biofilter and an electrooxidation cell

An attached-growth continuous flow hydrogenotrophic denitrification system was investigated for groundwater treatment. Two bench-scale packed-bed reactors were used in series, without external pH adjustment or carbon source addition, while inorganic carbonate salts already contained in the groundwater were the sole carbon source used by the denitrifying bacteria. The hydrogen was produced by water electrolysis using renewable energy sources thus minimizing resource-draining factors of the treatment process. The biofilter was subjected to a combination of three groundwater retention times (13.5, 27 and 54 min, corresponding to 20, 10 and 5 mL min−1 inlet water flow rates) and two hydrogen flow values (10 and 20 mL min−1) to evaluate its efficiency under different operating parameters. In all cases, significant nitrate percentage removals were achieved, ranged between 64.1% and 100%. The treatment process appears to slow down with lower retention times and H2 flow rate values, although residual nitrate concentrations were always in the range of 0–5.1 mg L−1, values below the maximum permitted limit of 11.3 mg L−1. In cases where nitrite accumulation was detected, a continuous flow electrochemical oxidation process with three different current density values (5.0, 7.5 and 10.0 mA cm−2) was examined as a post-treatment step aiming to completely remove the toxic nitrite anions. Finally, an advanced mathematical model of the attached growth hydrogenotrophic denitrification process was developed to predict concentrations of all the substrates examined in the bio-filter (nitrate, nitrite, inorganic carbon and hydrogen).

Nitrogen removal enhanced by its migration and transformation in a three-chamber microbial electrolysis cell

In this study, a three-chamber microbial electrolysis cell (MEC) was established for the simultaneous migration, transformation and removal of NH4+-N, NO2−-N, and NO3−-N under different applied voltages. The results showed that higher applied voltages accelerated the migration rate of NH4+-N while had little effect on NO2−-N and NO3−-N. The highest total nitrogen (TN) removal efficiency was achieved in the MEC system at 1.0 V (61.40 ± 19.58 %). At the optimal voltage of 1.0 V, nitrogen removal performance at different carbon to nitrogen (C/N) ratios was also discussed. Under different C/N ratios, TN removal efficiencies maintained stable, which suggested that the migration of nitrogen species by applied voltages combined with nitrogen transformation was beneficial for nitrogen removal in MEC systems. Also, different denitrification pathways could be influenced by reasonably allocating carbon sources. Meanwhile, the whole system was enriched with autotrophic and heterotrophic denitrifiers (e.g. Proteobacteria, Patescibacteria, Thermomonas, and Ferruginibacter), implying that heterotrophic denitrification, hydrogenotrophic denitrification and electroactive autotrophic denitrification coexisted and collaborated in this MEC system for nitrogen removal. The “migration-transformation-removal” model of nitrogen was innovatively proposed and meanwhile this proof-of concept study provided a new insight into nitrogen removal in low C/N ratio wastewater.

Development of a novel periodic venting-controlled membrane biofilm reactor for hydrogenotrophic denitrification: Process performance and microbial mechanism

The performance of anaerobic membrane biofilm reactor (MBfR) has long been underexploited due to the limited delivery of active gas to the biofilm on the close-end hollow fiber membranes (HFMs). Here, to establish a valid scheme for overcoming this obstacle, we coupled a denitrifying H2-based MBfR to an automatic venting unit for intermittently updating intramembrane gas, and a microsensor unit for investigating the H2 diffusion behavior. In short-term experiments, at the HFM length of 60 cm and H2 supplying pressure of 1.2 atm, the periodic venting strategy, in which the venting time and interval were set as 30 s and 17 min, respectively, was validated to be capable of improving the denitrification performance of H2-based MBfR. The results of long-term experiment show that compared to the conventional close-end operation, the around 50% increase in denitrification flux was achieved in the periodic venting-controlled system, which was found to be associated with the increased biofilm thickness and the evolution of denitrifying bacteria (DNB) and functional genes in the biofilm at the HFM distal end. Model outputs manifest that regardless of the HFM length, the biofilm regions with high DNB activity at the distal end were much narrower than those at the proximal end of the close-end HFMs; owing to the improved H2 delivery, the active biofilm region at the distal end of 60 cm HFM was significantly enlarged from 0–225 µm in the case of close-end operation to 0–450 µm in the venting-controlled system. The proposed periodic venting strategy opens a new avenue for the management of anaerobic MBfR.

Efficacy of a Fine Bubble Diffuser in Enhancing Attached Biofilm Hydrogenotrophic Denitrification Reactor Performance

The efficacy of a fine bubble (FB) diffuser in enhancing the performance of an attached biofilm hydrogenotrophic denitrification (HD) reactor was evaluated. HD reactors equipped with an FB diffuser (FB reactor) and an air stone (AS) diffuser that produced ordinary bubbles (AS reactor) were operated in parallel at different hydraulic retention times (HRTs) in a synthetic groundwater treatment experiment. A reduction in H2 consumption of approximately 77% was achieved using the FB diffuser to reach a gas-liquid mass transfer coefficient similar to that of the AS diffuser. The high gas dissolution efficiency of the FB diffuser resulted in an effective nitrogen removal rate (NRR) enhancement, requiring less H2 supply. The highest value of NRR at 53.0±9.8 g-N/m3/d was obtained in the FB reactor at a minimum HRT of 3 h, which was two-fold higher than the corresponding value from the AS reactor. The FB reactor also had the lowest requirement of H2 for denitrification reaching 0.1 m3-H2/g-N in this condition, which was ten-fold lower than that in the AS reactor. Furthermore, the suspended sludge concentration in the FB reactor was lower than that in the AS reactor, indicating that the application of the FB diffuser can minimize excess suspended sludge accumulation inside the HD reactor. Microbial community analysis showed the predominance of Thauera spp. reaching a relative abundance of 15.7–27.3% in the FB reactor, suggesting a contribution to the HD. This finding can provide insight into the application of the FB diffuser for optimizing nitrate-contaminated groundwater treatment technology by HD.

Limited dissolved oxygen facilitated nitrogen removal at biocathode during the hydrogenotrophic denitrification process using bioelectrochemical system

Effects of limited dissolved oxygen (DO) on hydrogenotrophic denitrification at biocathode was investigated using bioelectrochemical system. It was found that total nitrogen removal increased by 5.9%, as DO reached about 0.24 mg/L with the cathodic chamber unplugged (group R_Exposure). With the presence of limited DO, not only the nitrogen metabolic pathway was influenced, but the composition of microbial communities of ammonia-oxidizing bacteria and nitrite-oxidizing bacteria were enriched accordingly. After metagenomic analysis, enriched genes in R_Exposure were found to be associated with nearly each of nitrogen removal steps as denitrification, nitrification, DNRA, nitrate assimilation and even nitrogen fixation. Moreover, genes encoding both Complexes III and IV constituted the electron transfer chain were significantly enriched, indicating that more electrons would be orientated to the reduction of NO2–-N, NO-N and oxygen. Therefore, enhanced nitrogen removal could be attained through the co-respiration of nitrate and oxygen by means of NH4+-N oxidation.

Enhancement of iron-based nitrogen removal with an electric-magnetic field in an upflow microaerobic sludge reactor (UMSR).

Traditional denitrification often produces high operating costs and excessive sludge disposal expenses due to conventional carbon sources. A novel electric-magnetic field (MF) 48 mT with Fe0 and C-Fe0 powder in an upflow microaerobic sludge reactor (UMSR) improved nitrogen removal from wastewater without organic carbon resources and gave richness to the heterotrophic bacterial community. In the current study, the reactor was operated for 78 ± 2days, divided into five stages (without Fe0, with Fe0, coupling with MF, without coupling with MF, and coupling with MF again), at a hydraulic retention time (HRT) of 2.5h, with an influent loading of ammonium (NH4+-N) 50 ± 2mg/L, at 25-27°C, and less than 1.0mg/L dissolved oxygen (DO). The results demonstrated nitrogen removal efficiency enhanced after coupling with MF on the levels of NO3--N by 76% with an effluent concentration of 8.7mg/L, NH4+-N by 72% with an effluent concentration of 13.6mg/L, and total nitrogen removal (TN) by 76%, respectively. After coupling the MF with the reactor, the microbial community data analysis showed the dominant abundance of ammonia-oxidizing bacteria, heterotrophic nitrifying bacteria, and denitrifying bacteria on the level of Anaerolineaceae_uncultured 2%, which is capable of denitrification that uses Fe2+ as an electron source, Gemmatimonadaceae_uncultured 4%, Hydrogenophaga 4% which is capable of catalyzing hydrogenotrophic denitrification and correlating to nitrate removal, denitrification and desulfurization bacteria SBR1031_norank 18%, anammox-bacteria Saccharimonadales_norank 2%, and (AOM) Limnobacter 3% in the sludge.

Biological nitrogen removal from low carbon wastewater.

Nitrogen has traditionally been removed from wastewater by nitrification and denitrification processes, in which organic carbon has been used as an electron donor during denitrification. However, some wastewaters contain low concentrations of organic carbon, which may require external organic carbon supply, increasing treatment costs. As a result, processes such as partial nitrification/anammox (anaerobic ammonium oxidation) (PN/A), autotrophic denitrification, nitritation-denitritation and bioelectrochemical processes have been studied as possible alternatives, and are thus evaluated in this study based on process kinetics, applicability at large-scale and process configuration. Oxygen demand for nitritation-denitritation and PN/A is 25% and 60% lower than for nitrification/denitrification, respectively. In addition, PN/A process does not require organic carbon supply, while its supply for nitritation-denitritation is 40% less than for nitrification/denitrification. Both PN/A and nitritation-denitritation produce less sludge compared to nitrification/denitrification, which saves on sludge handling costs. Similarly, autotrophic denitrification generates less sludge compared to heterotrophic denitrification and could save on sludge handling costs. However, autotrophic denitrification driven by metallic ions, elemental sulfur (S) and its compounds could generate harmful chemicals. On the other hand, hydrogenotrophic denitrification can remove nitrogen completely without generation of harmful chemicals, but requires specialized equipment for generation and handling of hydrogen gas (H2), which complicates process configuration. Bioelectrochemical processes are limited by low kinetics and complicated process configuration. In sum, anammox-mediated processes represent the best alternative to nitrification/denitrification for nitrogen removal in low- and high-strength wastewaters.

Modelling of hydrogenotrophic denitrification process in a venturi-integrated membrane bioreactor

ABSTRACT The aim of this study is to model a hydrogenotrophic denitrification process in a venturi-integrated submerged membrane bioreactor (MBR) system. The MBR was operated in batch mode using feed concentrations of 100 and 150 mg NO3-N/L. In contrast to most of the denitrification process models that represent the mixed culture with one composite biomass parameter, the biomass was subdivided into two main categories in this modelling study: mainly nitrate-reducing biomass and mainly nitrite-reducing biomass. The determination coefficients (r 2) in the range of 0.97–0.99 indicate that the model successfully simulates the concentrations of nitrate- and nitrite-nitrogen in the bioreactor. The maximum specific growth rate of nitrite-reducing biomass (0.06 h−1) was found to be higher than that of nitrate-reducing biomass (0.0002 h−1). Similarly, the growth yield coefficient of nitrite-reducing biomass was higher than that of nitrate-reducing biomass (0.44 vs. 0.31 g biomass/g substrate). The kinetic and stoichiometric coefficients obtained from this modelling study suggest that the limiting step determining the overall conversion rate of hydrogenotrophic denitrification process is the conversion of nitrite to nitrogen gas.

The key factor in High Pressure Hydrogenotrophic Denitrification: Hydrogen partial pressure

In the envisaged hydrogen economy, H2 could be an interesting alternative electron donor for the denitrification of drinking water or wastewater. The main obstacle to engineering the hydrogenotrophic denitrification process is the low solubility of H2 in water under atmospheric pressure, which limits denitrification rate and nitrogen removal efficiency. In this paper, we demonstrated a novel configuration of hydrogenotrophic denitrification, namely High Pressure Hydrogenotrophic Denitrification (HPHD). Elevated H2 partial pressure (pH2) was employed to increase dissolved H2 concentration and concomitantly enhance denitrification rate. Our results showed that the specific denitrification rate increased from 9.6 mg N/(gVSS·h) at 0.5 bars to 51.0 mg N/(gVSS·h) at 9 bars in HPHD. The denitrification effect could be retained with elevated pH2 at a low temperature. The specific denitrification rate at 3 bars and 15 °C was 20.5 mg N/(gVSS·h), approximately 1.5 times that at 1 bar and 30 °C, which was quite beneficial for hydrogenotrophic denitrification under cold conditions. Different from nitrite reduction, less impact was observed on nitrate reduction by low temperature, which explained high nitrite accumulation in HPHD at 15 °C. Overall, our investigations shed light on the role of pH2 in the promising solution for nitrogen removal in HPHD.

Development and characterizations of hydrogenotrophic denitrification granular process: Nitrogen removal capacity and adaptability

Hydrogenotrophic denitrification (HD) is a promising autotrophic biological process for advanced nitrogen removal, while sludge granulation was seldom reported. This study aimed to cultivate granular sludge to improve capacity and stability of HD process. The resulting HD granular sludge performed high nitrogen removal rate (NRR) of 0.42 ± 0.0.4 kgN/(m3·d) with low accumulation of nitrite and nitrous oxide emission. HD granular sludge reactor performed over 3 times higher NRR compared to that in HD fixed-bed biofilm reactor (0.13 ± 0.01 kgN/(m3·d). Besides, granular sludge reactor could treat groundwater well even at the low temperature of 15 °C. The dominant genera were Hydrogenophaga and Comamonas in granular sludge, and Dechloromonas in biofilm. Noticeably, sulfate in the groundwater stimulated the growth of sulfur converting microbes with increasing abundances of sulfite reductase gene and sulfate-reducing bacteria Desulfovibrio. This study highlights the potential implementation of HD process in granular sludge reactor for advance nitrogen removal from impaired groundwater.

Rice husk-intensified cathode driving bioelectrochemical reactor for remediating nitrate-contaminated groundwater.

To achieve economical and eco-friendly denitrification, rice husk-intensified cathode driving bioelectrochemical reactor (RCBER) was constructed with rice husk as solid-phase carbon source and microbial carrier. Results demonstrated that the application of current improved the utilization of rice husk and enhanced the denitrification, and the quenching of anodic hydroxyl radicals by rice husk also improved the microbial resistance to current. The highest nitrate removal rate as 0.34 mg-N/(L∙d), higher economic benefits, i.e., current efficiency as 31.6% and energy consumption as 2.43 kWh/g NO3--N, and the highest environmental benefit, i.e., hydrogenotrophic denitrification contribution as 37.9%, were obtained at 200 mA/m2. The best performance at 200 mA/m2 was related to its better microenvironment, such as lower accumulation of anodic by-products and higher bioavailability of rice husks, as well as higher microbial metabolic activity, such as stable extracellular polymeric substance, the maximum electron transport system activity as 11.63 ± 0.14 μg O2·g-1·min-1·mg protein-1 and the highest activity of nitrate reductase (3.15-fold that of control check). The application of current realized the coexistence of heterotrophic and hydrogenotrophic denitrifiers, and multiple functional bacteria such as anaerobic denitrifiers Flavobacterium, aerobic denitrifiers Comamonas, hydrogenotrophic denitrifiers Thermomonas and electron transfer-related Enterobacter coexisted at 200 mA/m2, thereby improving RCBER's adaptability to the complex microenvironment. This study provides the theoretical basis for realizing a win-win situation of environmental pollution remediation and agricultural waste disposal.

Enhanced Cd(II) immobilization in sediment with zero-valent iron induced by hydrogenotrophic denitrification

In this study, an integrated system of Fe0 and hydrogenotrophic microbes mediated by nitrate (nitrate-mediated bio-Fe0, NMB-Fe0) was established to remediate Cd(II)-contaminated sediment. Solid phase characterization confirmed that aqueous Cd(II) (Cd(II)aq) was successfully immobilized and enriched on iron surface due to promoted iron corrosion driven by hydrogenotrophic denitrification and subsequent greater biomineral production such as magnetite, lepidocrocite and green rust. Compared to a Cd(II)aq removal of 21.1% in overlying water of the nitrate-mediated Fe0 (NM-Fe0) system, the NMB-Fe0 system obtained a much higher Cd(II)aq removal of 83.1% after 7 d remediation. The leaching test and sequential extraction results also showed that the leachability of Cd(II) decreased by 75.9% while the residual fraction of Cd(II) increased by 185.7% in comparison with untreated sediment. Besides, the Cd(II)aq removal raised with the increase of nitrate concentration and Fe0 dosage, further revealing the promotion effect of nitrate on Cd(II) removal by bio-Fe0. This study highlighted the involvement of bio-denitrification in the remediation of Cd(II)-contaminated sediment by Fe0 and provided a new insight to enhance its reactivity and applicability for Cd(II) immobilization.

Investigation of nitrite accumulation by hydrogenotrophic denitrification in a moving bed biofilm reactor for partial denitrification and anammox process.

The partial denitrification and anammox (PDA) process has received attention for its ability to optimize treatment of wastewater containing a low NH4+-N concentration. This study investigated the suitable operational conditions for NO2--N accumulation by hydrogenotrophic denitrification (HD) in operation of a laboratory-scale moving bed biofilm reactor, for future application in the PDA process. NO2--N accumulation was achieved by minimizing the H2 flow rate under optimized conditions (i.e., 15 mL/min H2 flow rate, 40 mg-N/L influent NO3--N, 7.0 h hydraulic retention time, and 2 L working volume). Hydrogenophaga comprised 39.2% of the bacterial abundance after NO2--N accumulated, indicating its contribution to the NO2--N accumulation. In addition, an intermittent H2 supply maintained the NO2--N accumulation rate (NAR) and maximized the nitrite accumulation efficiency (NAE). A H2 supply ratio of 0.7 (i.e., ON: 7 min, OFF: 3 min) was optimal, which induced increases in NAR, NAE, and the NO3--N removal efficiency that reached 0.07±0.01 kg-N/m3/d, 64.4±14.5%, and 89.2±8.9%, respectively. The ratio of H2 supply rate to the NO3--N loading rate was calculated as 4.3 in this experiment, which may represent the optimal balance for maximization of NO2--N accumulation by HD.

Bioelectrochemical reactor improved by assembling anode with rice husk for treating nitrate-contaminated groundwater

To maximize the hydrogenotrophic denitrification and minimize the negative impact of anode by-products, a rice husk-enhanced anode bioelectrochemical reactor (RABER) was constructed by assembling anode with rice husk, and the denitrification performances and mechanisms involved were investigated. It was demonstrated that rice husk can be used as a slow-release carbon source and microbial carrier in RABER and weakened the anodic by-products. RABER obtained the highest contribution of hydrogenotrophic denitrification as 33.7% and achieved the fastest nitrate removal rate of 0.27 mg-N/(L·d) at 200 mA/m2. The relative lower accumulation of dissolved oxygen (1.99 ± 0.11 mg/L) and hydroxyl radicals were obtained at 200 mA/m2. Moreover, stable extracellular polymeric substances and higher activities of denitrification-related enzymes, i.e., around 2.78-fold for nitrate reductase and 1.96-fold for nitrite reductase those of 0 mA/m2, promoted denitrification in RABER at 200 mA/m2. At an optimal current density of 200 mA/m2, the coexistence of cellulose-degrading bacteria (Cellvibrio), aerobic denitrifier (Brevundimonas), anaerobic denitrifier (Flavobacterium), and hydrogenotrophic denitrifier (Alicycliphilus) was achieved, and the inorganic carbon generated by heterotrophic denitrifiers and electrochemical mineralization was sufficient to meet the demands of hydrogenotrophic denitrifiers, conductive to the heterotrophic‑hydrogenotrophic synergistic denitrification. This study provided a theoretical basis for the enhancement of hydrogenotrophic denitrification in BER as well as minimizing the side effects of the anode and providing a new strategy for the disposal of agricultural waste.

Characterization of nitrous oxide and nitrite accumulation during iron (Fe(0))- and ferrous iron (Fe(II))-driven autotrophic denitrification: mechanisms, environmental impact factors and molecular microbial characterization

The iron (Fe(0))-/ferrous iron (Fe(II))-driven autotrophic denitrification processes have been alternative methods for nitrogen removal from low organic carbon (OC) wastewater, but the accumulation of nitrous oxide (N2O) and nitrite (NO2−) along with these processes remains unclear. This research aimed to systematically characterize the N2O/NO2− accumulation in Fe(0)-/Fe(II)-ADN processes through investigating the mechanisms, impact factors, and molecular biological characteristics. Results showed that Fe(II)-ADN was effective in NO3− reduction but was less efficient in N2O reduction (k = 0.50 h−1) than Fe(0)-ADN (k = 1.82 h−1). NO2−/N2O accumulation in Fe(II)-ADN (28.6%/30.7%) was much higher than that in Fe(0)-ADN (12.6%/1.5%). Introducing hydrogenotrophic denitrification (H-ADN) into Fe(II)-ADN system significantly (p < 0.05) reduced NO2−/N2O accumulation. Fe(0)-ADN was proved a coupled process of Fe(II)- and H-ADN by in-situ generating Fe(II)/H2, and Fe(II)-ADN and H-ADN mainly contributed to NO3− and NO2−/N2O reduction, respectively. Optimum pH (7.5) and temperature (30–35 °C) were confirmed with controlled NO2–/N2O accumulation and effective denitrification. Dosing inorganic carbon (IC) and OC enhanced denitrification and reduced NO2–/N2O accumulation, where OC was more efficient with an optimum dosage of 0.25 mmol C/mmol N. 16S rRNA high-throughput sequencing and Pearson Correlation Coefficients verified that Thiobacillus was the main contributor to NO3− reduction, whereas Thauera and Acidovorax possessed high NO2−/N2O reduction capability. Real-time quantitative polymerase chain reaction and enzyme activity assay demonstrated that the nitrite reductase encoded by gene nirK and the nitrous oxide reductase encoded by gene nosZ were efficient in catalyzing the further reduction of NO2− and N2O, respectively. This study could provide an in-depth understanding of NO2−/N2O accumulation in Fe(II)-/Fe(0)-ADN processes and contribute to their application, optimization and secondary pollutants control.

Column study of Cd(II) removal and longevity by nitrate-mediated zero-valent iron with mixed anaerobic microorganisms

In this study, hydrogen-autotrophic microorganisms and zero-valent iron (Fe0) were filled into columns to investigate hydrogenotrophic denitrification effect on cadmium (Cd(II)) removal and column life-span with sand, microorganisms, Fe0 and bio-Fe0 columns as controls. In terms of the experiment results, the nitrate-mediated bio-Fe0 column showed a slow Cd(II) migration rate of 0.04 cm/PV, while the values in the bio-Fe0 and Fe0 columns were 0.06 cm/PV and 0.14 cm/PV respectively, indicating much higher Cd(II) removal efficiency and longer service life of the nitrate-mediated bio-Fe0 column. The XRD and SEM-EDX results implied that this improvement was attributed to hydrogenotrophic denitrification that caused more serious iron corrosion and larger amount of secondary mineral generation (e.g., green rust, lepidocrocite and goethite). These active minerals provided more reaction sites for Cd(II) adsorption and further immobilization. In addition, the decrease of Cd(II) migration front and the increase of removal capacity along the bio-Fe0 column mediated by nitrate presented an uneven distribution in reactive zone. The latter half part was identified to be a more active region for Cd(II) immobilization. The above results indicate that the introduction of nitrate and microorganisms will improve the performance of iron-based permeable reactive barriers for the remediation of Cd(II)-containing groundwater.