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Dissolved methane is a hurdle for anaerobic wastewater treatment, which would be stripped into the atmosphere by conventional bubble aeration and increase the release of greenhouse gases into the environment. The high oxygen transfer efficiency and less turbulence in membrane aerated biofilm reactor (MABR) could prevent the stripping of dissolved methane. In this study, an MABR was established to remove dissolved methane aerobically in parallel to the nitrogen removal driven by the anammox process. The long-term results demonstrated that aerobic methane oxidation has a short start-up period, in which a high level (>90 %) of dissolved methane removal was achieved in 20 days. Meanwhile, the anammox-based nitrogen removal process reached a total nitrogen removal rate of ∼150 mg N/L/d (0.27 g N/m 2 /d). In situ batch tests confirmed the active bioreactions of ammonia-oxidizing bacteria, nitrite-oxidizing bacteria, anammox bacteria and aerobic methanotrophs, while 16S rRNA gene amplicon sequencing further validated their existence. Moreover, nitrite/nitrate-dependent anaerobic methane oxidation (n-DAMO) bacteria were enriched to a relative abundance of 2.5 % on Day 372, suggesting their potential role in removing nitrogen and dissolved methane in the MABR. This study provides an alternative technology for removing dissolved methane and nitrogen in parallel from anaerobically treated wastewater., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper. An author of this submission, Jianhua Guo, serves as an editor of Water Research. The manuscript has been handled and reviewed independently by other editors of the journal, with Jianhua Guo recusing themselves from any decisions related to the evaluation and acceptance of this work., (Copyright © 2024 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

Sewer systems are critical water infrastructures for sewage collection and transportation services but are frequently challenged by odour nuisance, corrosion and greenhouse gas emissions, primarily driven by sulfide and methane production. This study investigated the effectiveness of multifunctional nanoscale zero-valent iron (nZVI) in controlling sulfide and methane, along with its downstream impacts on wastewater treatment. Two continuous flow laboratory-scale reactor systems were used: sewer reactors and sequencing batch reactors (SBRs). Intermittent doses of 50 mg Fe/L of nZVI were introduced daily for a 6-h cycle in the experimental sewer reactors. Results indicated reduced sulfide (by 8.5±0.5 mg S/L during dosing; 4.2±0.6 mg S/L off-dosing) and methane (by 16.6±1.9 mg COD/L during dosing; 12.6±1.3 mg COD/L off-dosing) concentrations compared to the control. This reduction involved sulfide removal (0.12±0.01 g S/g Fe or 0.20±0.02 mol S/mol Fe) and the inhibition of microbial sulfate-reducing and methanogenic activities. Sulfate-reducing bacteria (SRB) and methanogenic archaea (MA) activities exhibited dynamic inhibition with long-term nZVI addition (SRB: 58 % after the first dose, 21 % after 3 months; MA: 27 % to 39 %). Additionally, the sewer-dosed nZVI improved downstream phosphorus removal (0.42±0.04 mg P/mg Fe or 0.76±0.07 mol P/mol Fe) and enhanced sludge settleability and dewaterability. These findings highlight the potential of intermittent nZVI dosing for effective sulfide and methane control in sewers while delivering downstream benefits for integrated urban wastewater management., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2025. Published by Elsevier Ltd.)

The anaerobic oxidation of methane (AOM) carried out by anaerobic methanotrophic archaea (ANME) plays an important role in mitigating methane emissions from aqueous environments and has applications in bioremediation and wastewater treatment. Previous studies showed that AOM could be coupled to chromate reduction. However, the specific responsible microorganisms and the biochemical mechanisms are unclear. Herein, we showed that a consortium dominated by ANME "Candidatus Methanoperedens" was able to couple AOM to the reduction of Cr(VI) to Cr(III) at a stoichiometry close to the theoretical ratio. Quantitative distribution analysis of Cr(III) products suggested Cr(VI) was predominantly reduced via the extracellular respiratory pathways. Further Cr(III)-targeted fluorescent visualization combined with single-cell electron microscopic imaging suggested that Cr(VI) was reduced by "Ca. Methanoperedens" independently. Biochemical mechanism investigation via proteomic analysis showed proteins for nitrate reduction under nitrate-reducing conditions were significantly downregulated in Cr(VI)-reducing incubation. Instead, many multiheme cytochrome c (MHCs) were among the most upregulated proteins during the Cr(VI) reduction process, suggesting MHC-governed pathways for extracellular Cr(VI) reduction. The significant upregulation of a formate-dependent nitrite reductase during Cr(VI) reduction indicated its potential contribution to the small proportion of Cr(VI) reduction inside cells., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

Single cell protein (SCP, or microbial protein) is one of the emerging alternative protein sources to address the global challenge of food insecurity. Recently, the SCP produced from methane has attracted substantial attention since methane is a renewable resource attainable from anaerobic digestion. However, the supply of methane, an insoluble gas in water, is one of the major challenges in producing methane-based SCP. This work developed a novel bioreactor configuration, in which hollow fiber membrane was used for efficient methane supply while microorganisms were growing in the suspended form favourable for the biomass harvest. Over a 312-day operation, the impacts of three critical parameters on the SCP production were investigated, including the ratio of methane loading to ammonium loading, the ratio of methane loading to oxygen loading, and the sludge retention time (SRT). Under the condition of 4 g CH 4 /g NH 4 + , 4 g O 2 /g CH 4 and 5.05 g SCP/g N, respectively. The protein content was up to 67 %, which is higher than the majority of reported values to date. Moreover, the methane and ammonium utilization efficiencies were both close to 100 %, suggesting the highly efficient utilization of substrates in this new bioreactor configuration. A high relative abundance of essential amino acids (EAA) above 42 % was achieved, representing the highest EAA content reported. These findings provide valuable insights into SCP production using methane as a feedstock.4 and 5.05 g SCP/g N, respectively. The protein content was up to 67 %, which is higher than the majority of reported values to date. Moreover, the methane and ammonium utilization efficiencies were both close to 100 %, suggesting the highly efficient utilization of substrates in this new bioreactor configuration. A high relative abundance of essential amino acids (EAA) above 42 % was achieved, representing the highest EAA content reported. These findings provide valuable insights into SCP production using methane as a feedstock., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Methane-dependent denitrification links the global nitrogen and methane cycles. Since its initial discovery in 2006, this process has been understood to involve a division of labor between an archaeal group and a bacterial group, which sequentially perform nitrate and nitrite reduction, respectively. Yao et al. have now revised this paradigm by identifying a Methylomirabilis bacterium capable of performing methane-dependent complete denitrification on its own., Competing Interests: Declaration of interests No interests are declared., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Microbial methane oxidation coupled to a selenate reduction process has been proposed as a promising solution to treat contaminated water, yet the underlying microbial mechanisms are still unclear. In this study, a novel methane-based membrane bioreactor system integrating hollow fiber membranes for efficient gas delivery and ultrafiltration membranes for biomass retention was established to successfully enrich abundant suspended cultures able to perform methane-dependent selenate reduction under oxygen-limiting conditions. The microbial metabolic mechanisms were then systematically investigated through a combination of short-term batch tests, DNA-based stable isotope probing (SIP) microcosm incubation, and high-throughput sequencing analyses of 16S rRNA gene and functional genes ( pmoA and narG ). We confirmed that the methane-supported selenate reduction process was accomplished by a microbial consortia consisting of type-II aerobic methanotrophs and several heterotrophic selenate reducers. The mass balance and validation tests on possible intermediates suggested that methane was partially oxidized into acetate under oxygen-limiting conditions, which was consumed as a carbon source for selenate-reducing bacteria. High-throughput 16S rRNA gene sequencing, DNA-SIP incubation with 13 CH 4 , and subsequent functional gene ( pmoA and narG ) sequencing results collectively proved that Methylocystis actively executed partial methane oxidation and Acidovorax and Denitratisoma were dominant selenate-reducing bacteria, thus forming a syntrophic partnership to drive selenate reduction. The findings not only advance our understanding of methane oxidation coupled to selenate reduction under oxygen-limiting conditions but also offer useful information on developing methane-based biotechnology for bioremediation of selenate-contaminated water.

The present study demonstrated that the perchlorate reduction rate in a methane-based membrane biofilm reactor was significantly enhanced from 14.4 to 25.6 mg-Cl/L/d by increasing copper concentration in the feeding medium from 1 to 10 μM, indicating a stimulatory effect of copper on the methane-supported perchlorate reduction process. Batch tests further confirmed that the increased copper concentration enhanced both methane oxidation and perchlorate reduction rates, which was supported by an increasing trend of functional genes (pmoA for methanotrophs and pcrA for specific perchlorate reducers) abundances through quantitative polymerase chain reaction (qPCR). Both 16S rRNA gene sequencing and functional genes (pmoA and pcrA) sequencing jointly revealed that the biofilm supplied with a higher copper concentration exhibited a more diverse microbial community. The methane-supported perchlorate reduction was accomplished through a synergistic association of methanotrophs (Methylocystis, Methylomonas, and Methylocystaceae) and perchlorate reducers (Dechloromonas, Azospira, Magnetospirillum, and Denitratisoma). Acetate may function as the key syntrophic linkage between methanotrophs and perchlorate reducers. It was proposed that the increased copper concentration improved the activity of particulate methane monooxygenase (pMMO) for methane oxidation or promoted the biosynthesis of intracellular carbon storage compounds polyhydroxybutyrate (PHB) in methanotrophs for generating more acetate available for perchlorate reduction., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Methane bioconversion to value-added liquid chemicals has been proposed as a promising solution to augment the petroleum-dominated chemical market. Recent investigations have reported that various electron acceptors (e.g., nitrite and nitrate) are available to drive methane bioconversion to short-chain fatty acids (SCFAs). However, little is known about effects of the rate electron acceptor supplied on liquid chemical production from methane. Herein, three independent membrane biofilm reactors (MBfRs) feeding with respective nitrate, nitrite, combined nitrate and nitrite were operated under high and low rate condition in succession, to study whether feeding rate of electron acceptors could impact the methane bioconversion to SCFAs and the associated microbiological features. Long-term operation showed that all tested electron acceptors with a high supply rate were favorable for methane bioconversion to SCFAs (990.9 mg L -1 d -1 , 1695.7 mg L -1 d -1 , and 2425.7 mg L -1 d -1 ), while under a low electron acceptor feeding rate, the SCFA production rate decreased to 8.9 mg L -1 d -1 , 16.8 mg L -1 d -1 , and 260.1 mg L -1 d -1 , respectively. Microbial community characterization showed that the biofilm was predominated by Methanosarcina, Methanobacterium, Propionispora and Clostridium. On the basis of the known metabolism characteristics of these microorganisms, it was assumed that these methanogens and fermenters contributed jointly to methane bioconversion to SCFAs. The findings could be helpful to understand the role of electron acceptor rate in methane bioconversion to liquid chemicals., Competing Interests: Declaration of Competing Interest None., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Although methane (CH 4 ) has been proven to be able to serve as an electron donor for bio-reducing various oxidized contaminants (e.g., selenate (SeO 4 )), little is known regarding the roles of oxygen in methane-based reduction processes. Here, a methane-based membrane biofilm reactor (MBfR) was established for evaluating the effects of oxygen supply rates on selenate reduction performance and microbial communities. The oxygen supply rate played a dual role (stimulatory or suppressive effect) in selenate reduction rates, depending on the presence or absence of dissolved oxygen (DO). Specifically, selenate reduction rate was substantially enhanced when an appropriate oxygen rate (e.g., 12 to 184 mg/L 2- )), little is known regarding the roles of oxygen in methane-based reduction processes. Here, a methane-based membrane biofilm reactor (MBfR) was established for evaluating the effects of oxygen supply rates on selenate reduction performance and microbial communities. The oxygen supply rate played a dual role (stimulatory or suppressive effect) in selenate reduction rates, depending on the presence or absence of dissolved oxygen (DO). Specifically, selenate reduction rate was substantially enhanced when an appropriate oxygen rate (e.g., 12 to 184 mg/L . d in this study) was supplied but with negligible DO. The highest selenate reduction rate (up to 34 mg-Se/L . d) was obtained under an oxygen supply rate of 184 mg/L . d. In contrast, excessive oxygen supply rate (626 mg/L . d) would significantly suppress selenate reduction rate under DO level of 3 mg/L. Accordingly, though the high oxygen supply rate (626 mg/L . d) would promote the expression of pmoA (5.9 × 10 9 copies g -1 ), the expression level of narG (a recognized gene to mediate selenate reduction) would be significantly downregulated (6.1 × 10 9 copies g -1 ), thus suppressing selenate reduction. In contrast, the expression of narG gene significantly increased to 2.8 × 10 10 copies g -1 , and the expression of pmoA gene could still maintain at 1.1 × 10 9 copies g -1 under an oxygen supply rate of 184 mg/L . d. High-throughput sequencing targeting 16S rRNA gene, pmoA, and narG collectively suggested Methylocystis acts as the major aerobic methanotroph, in synergy with Arthrobacter and Variovorax which likely jointly reduce selenate to selenite (SeO 3 ). Methylocystis was predominant in the biofilm regardless of variations of oxygen supply rates, while Arthrobacter and Variovorax were sensitive to oxygen fluctuation. These findings provide insights into the effects of oxygen on methane-dependent selenate reduction and suggest that it is feasible to achieve a higher selenate removal by regulating oxygen supply rates.2- ), and further to elemental selenium (Se 0 ). Methylocystis was predominant in the biofilm regardless of variations of oxygen supply rates, while Arthrobacter and Variovorax were sensitive to oxygen fluctuation. These findings provide insights into the effects of oxygen on methane-dependent selenate reduction and suggest that it is feasible to achieve a higher selenate removal by regulating oxygen supply rates., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2021. Published by Elsevier Ltd.)

Bioconversion of biogas to value-added liquids has received increasing attention over the years. However, many biological processes are restricted under acidic conditions owing to the excessive carbon dioxide (CO 2 , 30-40% v/v) in biogas. Here, using an enriched culture dominated by acid-tolerant ammonia-oxidizing bacteria (AOB) 'Candidatus Nitrosoglobus', this study examined the feasibility of producing methanol from methane in the CO 2 -acidified environment (i.e. pH of 5.0). Within the tested dissolved methane range (0.1-0.9 mM), methane oxidation by the acid-tolerant AOB culture followed first-order kinetics, with the same rate constant (i.e. 0.43 (L/(g VSS‧h)) between pH 7.0 and 5.0. The acidic methane oxidation showed robustness against high dissolved concentrations of CO 2 (up to 4.06 mM) and hydrogen sulfide (H 2 S up to 0.11 mM), which led to a high methanol yield of about 30-40%. As such, the raw biogas containing toxic CO 2 and H 2 S can directly serve for methanol production by this acid-tolerant AOB culture, economizing a conventionally costly biogas upgradation process. Afterwards, two batch reactors fed with methane and oxygen intermittently both obtained a final concentration of 1.5 mM CH 3 OH (equal to 72 mg chemical oxygen demand/L) in the liquid, suggesting it is a useful carbon source to enhance denitrification in wastewater treatment systems. In addition, ammonia availability was identified to be critical for a higher rate of this AOB-mediated methanol production. Overall, our results for the first time demonstrated the capability of a novel acid-tolerant AOB culture to oxidize methane, and also illustrated the technical feasibility to utilize raw biogas for methanol production at acidic conditions., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2021. Published by Elsevier Ltd.)

Being an energetic fuel, methane is able to support microbial growth and drive the reduction of various electron acceptors. These acceptors include a broad range of oxidized contaminants (e.g., nitrate, nitrite, perchlorate, bromate, selenate, chromate, antimonate and vanadate) that are ubiquitously detected in water environments and pose threats to human and ecological health. Using methane as electron donor to biologically reduce these contaminants into nontoxic forms is a promising solution to remediate polluted water, considering that methane is a widely available and inexpensive electron donor. The understanding of methane-based biological reduction processes and the responsible microorganisms has grown in the past decade. This review summarizes the fundamentals of metabolic pathways and microorganisms mediating microbial methane oxidation. Experimental demonstrations of methane as an electron donor to remove oxidized contaminants are summarized, compared, and evaluated. Finally, the review identifies opportunities and unsolved questions that deserve future explorations for broadening understanding of methane oxidation and promoting its practical applications., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2021. Published by Elsevier Ltd.)

Granular sludge exhibits unique features, including rapid settling velocity, high loading rate and relative insensitivity against inhibitors, thus being a favorable platform for the cultivation of slow-growing and vulnerable microorganisms, such as anaerobic ammonium oxidation (anammox) bacteria and nitrite/nitrate-dependent anaerobic methane oxidation (n-DAMO) microorganisms. While anammox granules have been widely applied, little is known about how to speed up the granulation process of n-DAMO microorganisms, which grow even slower than anammox bacteria. In this study, we used mature anammox granules as biotic carriers to embed n-DAMO microorganisms, which obtained combined anammox + n-DAMO granules within 6 months. The results of whole-granule 16S rRNA gene amplicon sequencing showed the coexistence of anammox bacteria, n-DAMO bacteria and n-DAMO archaea. The microbial stratification along granule radius was further elucidated by cryosection-16S rRNA gene amplicon sequencing, showing the dominance of n-DAMO archaea and anammox bacteria at inner and outer layers, respectively. Moreover, the images of cryosection-fluorescence in situ hybridization (FISH) verified this stratification and also indicated a shift in microbial stratification. Specifically, n-DAMO bacteria and n-DAMO archaea attached to the anammox granule surface initially, which moved to the inner layer after 4-months operation. On the basis of combined anammox + n-DAMO granules, a practically useful nitrogen removal rate (1.0 kg N/m 3 /d) was obtained from sidestream wastewater, which provides new avenue to remove nitrogen from wastewater using methane as carbon source., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Hydrogen and methane commonly co-exist in aquifer. Either hydrogen or methane has been individually utilized as electron donor for bio-reducing chromate. However, little is known whether microbial chromate reduction would be suppressed or promoted when both hydrogen and methane are simultaneously supplied as joint electron donors. This study for the first time demonstrated microbial chromate reduction rate could be accelerated by both hydrogen and methane donating electrons. The maximum chromate reduction rate (4.70 ± 0.03 mg/L·d) with a volume ratio of hydrogen to methane at 1:1 was significantly higher than that with pure hydrogen (2.53 ± 0.02 mg/L·d) or pure methane (2.01 ± 0.02 mg/L·d) as the sole electron donor (p < 0.01). High-throughput 16S rRNA gene amplicon sequencing detected potential chromate reducers (e.g., Spirochaetaceae, Delftia and Azonexus) and hydrogenotrophic bacteria (e.g., Acetoanaerobium) and methane-metabolizing microorganisms (e.g., Methanobacterium), indicating that these microorganisms might play important roles on microbial chromate reduction using both hydrogen and methane as electron donors. Abundant hupL and mcrA genes responsible for hydrogen oxidation and methane conversion were harbored, together with chrA gene for chromate reduction. More abundant extracellular cytochrome c and intracellular NADH were detected with joint electron donors, suggesting more active electron transfers., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Anaerobic technologies have been proposed as a promising solution to enhance bioenergy recovery and to transform a wastewater treatment plant (WWTP) from an energy consumer to an energy exporter. However, 20-60% of the methane produced remains dissolved in the anaerobically treated effluent, which is a potent greenhouse gas and is easily stripped out in the aeration tank. This study aims to develop a solution using dissolved methane to support denitrification, thus simultaneously enhancing nitrogen removal and achieving beneficial use of dissolved methane. By coupling anaerobic ammonium oxidation (anammox) with nitrite/nitrate-dependent anaerobic methane oxidation (n-DAMO), up to 85% of dissolved methane and more than 99% of nitrogen were removed in parallel in a biofilm system. Mass balance was conducted during both long-term operation and short-term batch tests, which indicated that n-DAMO bacteria and n-DAMO archaea indeed contributed jointly to the methane removal. The 16S rRNA gene amplicon sequencing further showed the co-presence of n-DAMO bacteria and n-DAMO archaea, while anammox bacteria were detected with a low relative abundance. This proposed technology can potentially be applied to reduce the carbon footprint and to save the organic carbon consumption in WWTPs.

An innovative process coupling anaerobic ammonium oxidation (anammox) with nitrite/nitrate-dependent anaerobic methane oxidation (n-DAMO) in membrane biofilm reactors (MBfRs) has been developed to achieve high-level nitrogen removal from both sidestream (i.e., anaerobic digestion liquor) and mainstream (i.e., domestic strength) wastewater. In this study, a 1D biofilm model embedding the n-DAMO and anammox reactions was developed to facilitate further understanding of the process and its optimization. The model was calibrated and validated using comprehensive data sets from two independent MBfRs, treating sidestream- and mainstream-strength wastewater, respectively. Modelling results revealed a unique biofilm stratification. While anammox bacteria dominated throughout the biofilm, n-DAMO archaea (coupling nitrate reduction with anaerobic methane oxidation) only occurred at the inner layer and n-DAMO bacteria (coupling nitrite reduction with anaerobic methane oxidation) spread more evenly with a slightly higher fraction in the outer layer. The established MBfRs were robust against dynamic influent flowrates and nitrite/ammonium ratios. Thicker biofilms were beneficial for not only the total nitrogen (TN) removal but also the system robustness. Additionally, a positive correlation between the nitrogen removal efficiency and the residual methane emission was observed, as a result of higher methane partial pressure required. However, there was a threshold of methane partial pressure, above which the residual methane increased but nitrogen removal efficiency was stable. Meanwhile, thicker biofilms were also favorable to achieve less residual methane emission. Simulation results also suggested the feasibility of methane-based MBfRs to polish mainstream anammox effluent to meet a stringent N discharge standard (e.g., TN < 5 mg/L)., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 The Authors. Published by Elsevier Ltd.. All rights reserved.)

The mainstream anaerobic ammonium oxidation (anammox) process provides strong support to the on-going paradigm shift from energy-negative to energy-neutral in wastewater treatment plants. However, the low temperature (e.g., below 15 °C) represents one of the major challenges for mainstream anammox in practice. In this study, a stable nitrogen removal rate (0.13 kg m -3 day -1 ), together with a high-level effluent quality (<5.0 mg N L -1 ), was achieved in a lab-scale upflow membrane biofilm reactor (MBfR) by coupling anammox with nitrite/nitrate-dependent anaerobic methane oxidation (n-DAMO) microorganisms, at a temperature as low as 10 °C. With the temperature being progressively decreased from 25 to 10 °C, the total nitrogen removal efficiency was maintained in the range of 90-94% at a constant hydraulic retention time of 9 h. The impact of temperature on the biofilm system coupling anammox and n-DAMO reactions increased at a lower temperature range with higher Arrhenius coefficients. Additionally, 16S rRNA gene sequencing results showed that anammox bacteria, n-DAMO bacteria, and n-DAMO archaea jointly dominated the biofilm, and their respective abundances remained relatively stable when the temperature was decreased. The major reason for this temperature-tolerated performance is the overcapacity developed, which is indicated by biofilm thickness measurements and mathematical modeling. The stable performance obtained in this study shows promise for the n-DAMO application in domestic wastewater.

While the anaerobic ammonium oxidation (anammox) process has been applied for nitrogen removal from high-strength wastewater, nitrate accumulation in effluent still represents a major concern. Here, a novel process, named the one-stage PNAM, that integrates the Partial Nitritation (PN), Anammox and Methane-dependent nitrite/nitrate reduction reactions in a single membrane biofilm reactor (MBfR) is developed. With feeding of 1030 mg NH4+-N/L at a hydraulic retention time of 16 h, the proposed one-stage PNAM process achieved an average total nitrogen removal efficiency of 98% and a nitrogen removal rate of 1.5 kg N/m3/d (1.4-1.8 g N/m2/d) by using methane as the sole carbon-based electron donor. The N2O emission was determined to be 0.34% ± 0.01%. Microbial community characterization revealed that ammonia-oxidizing bacteria (AOB), anammox bacteria, nitrite/nitrate-dependent anaerobic methane oxidation (n-DAMO) bacteria and archaea co-developed in the biofilm. Batch tests showed that AOB, anammox bacteria and n-DAMO microorganisms were indeed jointly responsible for the nitrogen removal. This one-stage PNAM process can potentially be applied to treating high-strength wastewater, such as anaerobic sludge digestion liquor or landfill leachate., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Given our vast methane reserves and the forecasted shortage of crude oil in the not too distant future, the conversion of methane into value-added liquid chemicals or fuels would be beneficial. The generated chemicals or fuels could augment the petroleum-dominated chemical market, and also satisfy the increasing demand for transportation fuels. While methane bioconversion to liquid chemicals has just been reported recently, there is limited understanding of the process. This study aims to clarify the potential electron acceptors that could support the process. Here we operated four membrane biofilm reactors (MBfRs) fed with nitrate, nitrite, oxygen at a relatively low rate, and oxygen at a relatively high rate, respectively, to study if they can support methane bioconversion to short-chain fatty acids (SCFAs) and the associated microbiological features. All tested electron acceptors facilitated methane bioconversion to SCFAs (ranging from 1.1 to 36.7 mg acetate L -1 d -1 , or 3.4 to 114.6 mg acetate d -1 m -2 of biofilm). The carbon efficiency was estimated to be 7.9 ± 1.4% to 148.5 ± 1.3%, with an efficiency higher than 100%, suggesting the assimilation of other carbon, very likely CO 2 , into the products. A low oxygen supply rate of 46.4 ± 2.3 mg O 2 d -1 m -2 was found to be the most favorable among all the electron conditions provided according to the SCFAs production rate and also the carbon utilization efficiency. Microbial characterization revealed that completely different communities evolved in the respective reactors, suggesting diverse microbial pathways exist for methane bioconversion into value-added chemicals.

Due to serious eutrophication in water bodies, nitrogen removal has become a critical stage for wastewater treatment plants (WWTPs) over past decades. Conventional biological nitrogen removal processes are based on nitrification and denitrification (N/DN), and are suffering from several major drawbacks, including substantial aeration consumption, high fugitive greenhouse gas emissions, a requirement for external carbon sources, excessive sludge production and low energy recovery efficiency, and thus unable to satisfy the escalating public needs. Recently, the discovery of anaerobic ammonium oxidation (anammox) bacteria has promoted an update of conventional N/DN-based processes to autotrophic nitrogen removal. However, the application of anammox to treat domestic wastewater has been hindered mainly by unsatisfactory effluent quality with nitrogen removal efficiency below 80%. The discovery of nitrate/nitrite-dependent anaerobic methane oxidation (n-DAMO) during the last decade has provided new opportunities to remove this barrier and to achieve a robust system with high-level nitrogen removal from municipal wastewater, by utilizing methane as an alternative carbon source. In the present review, opportunities and challenges for nitrate/nitrite-dependent anaerobic methane oxidation are discussed. Particularly, the prospective technologies driven by the cooperation of anammox and n-DAMO microorganisms are put forward based on previous experimental and modeling studies. Finally, a novel WWTP system acting as an energy exporter is delineated.

There is great interest in microbial conversion of methane, an abundant resource, into valuable liquid chemicals. While aerobic bioconversion of methane to liquid chemicals has been reported, studies of anaerobic methane bioconversion to liquid chemicals are rare. Here we show that a microbial culture dominated by Candidatus 'Methanoperedens nitroreducens', an anaerobic methanotrophic archaeon, anaerobically oxidizes methane to produce acetate, indirectly via reaction intermediate(s), when nitrate or nitrite is supplied as an electron acceptor under a rate-limiting condition. Isotopic labeling tests showed that acetate was produced from certain intracellular storage compounds that originated from methane. Fluorescence in situ hybridization and Nile red staining demonstrated that polyhydroxyalkanoate in M. nitroreducens was likely one of the intracellular storage compounds for acetate production, along with glycogen. Acetate is a common substrate for the production of more valuable chemicals. The microbial conversion discovered in this study potentially enables a new approach to the use of methane as a feedstock for the chemical market.

Recently it has been shown that Candidatus 'Methanoperedens nitroreducens', an anaerobic methanotrophic archaea (ANME), can reduce nitrate to nitrite using electrons derived from anaerobic oxidation of methane. In this study, the growth kinetics of 'M. nitroreducens' enriched in a laboratory reactor were studied. In the experimental concentration range (up to 16 mg CH 4  L -1 ), anaerobic oxidation of methane by 'M. nitroreducens' was found to comply with first order kinetic model with a rate constant of 0.019 ± 0.006 h -1 and a biomass-specific rate constant of 0.04-0.14 L h -1  g -1 VSS. Meanwhile, the nitrate reduction to nitrite was well described by the Monod-type kinetic model with an affinity constant for nitrate of 2.1 ± 0.4 mg N L -1 , which is slightly higher than, but comparable to, that of most known denitrifying bacteria. This is the first time that the growth kinetics of 'M. nitroreducens' have been experimentally studied. The applicability of the kinetic model reported herein to this organism or similar organisms in natural or engineering systems requires further investigation., (Copyright © 2018. Published by Elsevier B.V.)

Microbially mediated anaerobic oxidation of methane (AOM) is a key process in the regulation of methane emissions to the atmosphere. Iron can serve as an electron acceptor for AOM, and it has been suggested that Fe(III)-dependent AOM potentially comprises a major global methane sink. Although it has been proposed that anaerobic methanotrophic (ANME) archaea can facilitate this process, their active metabolic pathways have not been confirmed. Here we report the enrichment and characterisation of a novel archaeon in a laboratory-scale bioreactor fed with Fe(III) oxide (ferrihydrite) and methane. Long-term performance data, in conjunction with the 13 C- and 57 Fe-labelling batch experiments, demonstrated that AOM was coupled to Fe(III) reduction to Fe(II) in this bioreactor. Metagenomic analysis showed that this archaeon belongs to a novel genus within family Candidatus Methanoperedenaceae, and possesses genes encoding the "reverse methanogenesis" pathway, as well as multi-heme c-type cytochromes which are hypothesised to facilitate dissimilatory Fe(III) reduction. Metatranscriptomic analysis revealed upregulation of these genes, supporting that this archaeon can independently mediate AOM using Fe(III) as the terminal electron acceptor. We propose the name Candidatus "Methanoperedens ferrireducens" for this microorganism. The potential role of "M. ferrireducens" in linking the carbon and iron cycles in environments rich in methane and iron should be investigated in future research.

Anaerobic oxidation of methane (AOM) plays a crucial role in controlling the flux of methane from anoxic environments. Sulfate-, nitrite-, nitrate-, and iron-dependent methane oxidation processes have been considered to be responsible for the AOM activities in anoxic niches. We report that nitrate-reducing AOM microorganisms, enriched in a membrane biofilm bioreactor, are able to couple selenate reduction to AOM. According to ion chromatography, X-ray photoelectron spectroscopy, and long-term bioreactor performance, we reveal that soluble selenate was reduced to nanoparticle elemental selenium. High-throughput 16S rRNA gene sequencing indicates that Candidatus Methanoperedens and Candidatus Methylomirabilis remained the only known methane-oxidizing microorganisms after nitrate was switched to selenate, suggesting that these organisms could couple anaerobic methane oxidation to selenate reduction. Our findings suggest a possible link between the biogeochemical selenium and methane cycles.

The discovery of denitrifying anaerobic methane oxidation (DAMO) has not only improved our understanding of global methane and nitrogen cycles, but also provided new technology options for removal of nitrate from nitrate-contaminated water. Previous studies have demonstrated DAMO organisms could remove nitrate and nitrite from wastewater under strictly anaerobically conditions. In the study, we investigate the feasibility of nitrate removal from groundwater, which contains dissolved oxygen in addition to nitrate. A membrane biofilm reactor (MBfR), inoculated with DAMO co-culture, was capable of treating synthetic groundwater containing highly contaminated nitrate (50  mg N/L) and oxygen (7-9 mg O 2 /L), with a maximum volumetric nitrate removal rate of 45 mg N/L-d. Accumulations of acetate and propionate were observed in some transient periods, indicating the possible involvement of acetate and propionate as intermediates in methane oxidation. The 16 S rRNA gene amplicon sequencing revealed that Candidatus Methylomirabilis, a known bacterial DAMO organism able to couple nitrite reduction with anaerobic oxidation of methane (AOM), was the dominant population. No archaeal DAMO organisms that are capable of coupling nitrate to AOM were observed, however, considerable amount of denitrifiers were developed in this system. Based on known metabolisms of these microorganisms and a series of batch studies, it was assumed that methane was oxidized into volatile fatty acids (VFAs) under oxygen-limiting conditions, then the generated VFAs served as carbon sources for these heterotrophic denitrifiers to remove nitrate. This study offers a potential technology for nitrate removal from groundwater by DAMO process in MBfR., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

The discovery of denitrifying anaerobic methane oxidation (DAMO) processes enables the complete nitrogen removal from wastewater by utilizing the methane produced on site from anaerobic digesters. This model-based study investigated the mechanisms and operational window for efficient nitrogen removal by coupling nitritation-anaerobic ammonium oxidation (Anammox) and methane-dependent denitrification in membrane biofilm reactors (MBfRs). A mathematical model was applied to describe the microbial interactions among Anammox bacteria, DAMO archaea, and DAMO bacteria. The model sufficiently described the batch experimental data from an MBfR containing an Anammox-DAMO biofilm with different feeding nitrogen compositions, which confirmed the validity of the model. The effects of process parameters on the system performance and microbial community structure could therefore be reliably evaluated. The impacts of nitritation produced NO2(-)/NH4(+) ratio, methane supply, biofilm thickness and total nitrogen (TN) surface loading were comprehensively investigated with the model. Results showed that the optimum NO2(-)/NH4(+) ratio produced from nitritation for the Anammox-DAMO biofilm system was around 1.0 in order to achieve the maximum TN removal (over 99.0%), independent on TN surface loading. The corresponding optimal methane supply increased while the associated methane utilization efficiency decreased with the increase of TN surface loading. The cooperation between DAMO organisms and Anammox bacteria played the key role in the TN removal. Based on these results, the proof-of-concept feasibility of a single-stage MBfR coupling nitritation-Anammox-DAMO for complete nitrogen removal was also tested through integrating the model with ammonia-oxidizing bacteria (AOB) and nitrite-oxidizing bacteria (NOB) processes whilst controlling the dissolved oxygen (DO) concentration in the simulated system. The maximum TN removal was found to be achieved at the bulk DO concentration of around 0.17 g m(-3) under the simulation conditions, with the AOB, Anammox bacteria and DAMO organisms coexisting in the biofilm., (© 2015 Wiley Periodicals, Inc.)

Methane in biogas has been proposed to be an electron donor to facilitate complete nitrogen removal using denitrifying anaerobic methane oxidizing (DAMO) microorganisms in an anaerobic ammonium oxidation (anammox) reactor, by reducing the nitrate produced. However, the slow growth and the low activity of DAMO microorganisms cast a serious doubt about the practical usefulness of such a process. In this study, a previously established lab-scale membrane biofilm reactor (MBfR), with biofilms consisting of a coculture of DAMO and anammox microorganisms, was operated to answer if the DAMO reactor can achieve a nitrate reduction rate that can potentially be applied for wastewater treatment. Through progressively increasing nitrate and ammonium loading rates to the reactor, a nitrate removal rate of 684 ± 10 mg-N L(-1) d(-1) was achieved after 453 days of operation. This rate is, to our knowledge, by far the highest reported for DAMO reactors, and far exceeds what is predicted to be required for nitrate removal in a sidestream (5.6-135 mg-N L(-1) d(-1)) or mainstream anammox reactor (3.2-124 mg-N L(-1) d(-1)). Mass balance analysis showed that the nitrite produced by nitrate reduction was jointly reduced by anammox bacteria at a rate of 354 ± 3 mg-N L(-1) d(-1), accompanied by an ammonium removal rate of 268 ± 2 mg-N L(-1) d(-1), and DAMO bacteria at a rate of 330 ± 9 mg-N L(-1) d(-1). This study shows that the nitrate reduction rate achieved by the DAMO process can be high enough for removing nitrate produced by anammox process, which would enable complete nitrogen removal from wastewater., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

The presence of a high level of dissolved methane (e.g., 20-26 g m(-3)) in the anaerobic sludge digestion liquor represents a major challenge to the treatment of this stream, as its emission to the atmosphere contributes significantly to the carbon footprint of wastewater treatment. Here we propose a new approach to simultaneous ammonium and dissolved methane removal from the anaerobic digestion liquor through integrating partial nitritation-Anammox and denitrifying anaerobic methane oxidation (DAMO) processes in a single-stage membrane biofilm reactor (MBfR). In such an MBfR, the anaerobic digestion liquor is provided in the bulk liquid, while oxygen is supplied through gas-permeable membranes to avoid dissolved methane stripping. A previously developed model with appropriate extensions was applied to assess the system performance under different operational conditions and the corresponding microbial interactions. Both influent surface loading (or hydraulic retention time) and oxygen surface loading are found to significantly influence the total nitrogen (TN) and dissolved methane removal, which jointly determine the overall system performance. The counter diffusion and concentration gradients of substrates cause microbial stratification in the biofilm, where ammonia-oxidizing bacteria (AOB) attach close to the membrane surface (biofilm base) where oxygen and ammonium are available, while Anammox and DAMO microorganisms jointly grow in the biofilm layer close to the bulk liquid where methane, ammonium, and nitrite are available with the latter produced by AOB. These results provide first insights and useful information for the design and operation of this new technology for simultaneous ammonium and dissolved methane removal in its potential future applications., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

Background: Anaerobic digestion has been widely applied to treat the waste activated sludge from biological wastewater treatment and produce methane for biofuel, which has been one of the most efficient solutions to both energy crisis and environmental pollution challenges. Anaerobic digestion sludge contains highly complex microbial communities, which play crucial roles in sludge treatment. However, traditional approaches based on 16S rRNA amplification or fluorescent in situ hybridization cannot completely reveal the whole microbial community structure due to the extremely high complexity of the involved communities. In this sense, the next-generation high-throughput sequencing provides a powerful tool for dissecting microbial community structure and methane-producing pathways in anaerobic digestion., Results: In this work, the metagenomic sequencing was used to characterize microbial community structure of the anaerobic digestion sludge from a full-scale municipal wastewater treatment plant. Over 3.0 gigabases of metagenomic sequence data were generated with the Illumina HiSeq 2000 platform. Taxonomic analysis by MG-RAST server indicated that overall bacteria were dominant (~93%) whereas a considerable abundance of archaea (~6%) were also detected in the anaerobic digestion sludge. The most abundant bacterial populations were found to be Proteobacteria, Firmicutes, Bacteroidetes, and Actinobacteria. Key microorganisms and related pathways involved in methanogenesis were further revealed. The dominant proliferation of Methanosaeta and Methanosarcina, together with the functional affiliation of enzymes-encoding genes (acetate kinase (AckA), phosphate acetyltransferase (PTA), and acetyl-CoA synthetase (ACSS)), suggested that the acetoclastic methanogenesis is the dominant methanogenesis pathway in the full-scale anaerobic digester., Conclusions: In short, the metagenomic sequencing study of this work successfully dissected the detail microbial community structure and the dominated methane-producing pathways of a full-scale anaerobic digester. The knowledge garnered would facilitate to develop more efficient full-scale anaerobic digestion systems to achieve high-rate waste sludge treatment and methane production.

Nitrogen removal by using the synergy of denitrifying anaerobic methane oxidation (DAMO) and anaerobic ammonium oxidation (Anammox) microorganisms in a membrane biofilm reactor (MBfR) has previously been demonstrated experimentally. In this work, a mathematical model is developed to describe the simultaneous anaerobic methane and ammonium oxidation by DAMO and Anammox microorganisms in an MBfR for the first time. In this model, DAMO archaea convert nitrate, both externally fed and/or produced by Anammox, to nitrite, with methane as the electron donor. Anammox and DAMO bacteria jointly remove the nitrite fed/produced, with ammonium and methane as the electron donor, respectively. The model is successfully calibrated and validated using the long-term (over 400 days) dynamic experimental data from the MBfR, as well as two independent batch tests at different operational stages of the MBfR. The model satisfactorily describes the methane oxidation and nitrogen conversion data from the system. Modeling results show the concentration gradients of methane and nitrogen would cause stratification of the biofilm, where Anammox bacteria mainly grow in the biofilm layer close to the bulk liquid and DAMO organisms attach close to the membrane surface. The low surface methane loadings result in a low fraction of DAMO microorganisms, but the high surface methane loadings would lead to overgrowth of DAMO bacteria, which would compete with Anammox for nitrite and decrease the fraction of Anammox bacteria. The results suggest an optimal methane supply under the given condition should be applied not only to benefit the nitrogen removal but also to avoid potential methane emissions.

The discovery of denitrifying anaerobic methane oxidation with nitrite as electron acceptor mediated by ' C andidatus Methylomirabilis oxyfera' connected the biogeochemical carbon and nitrogen cycle in a new way. However, it is important to have a comprehensive understanding about the distribution of M . oxyfera-like bacteria in the terrestrial realm, especially the wetland ecosystems that are known as the largest natural source of atmospheric methane. Here, our molecular evidence demonstrated that a wide geographical distribution of M . oxyfera-like bacteria at oxic/anoxic interfaces of various wetlands ( n = 91) over the Chinese territory. Intriguingly, the M . oxyfera-like bacteria were detected in some extreme environments, indicating that M . oxyfera-like bacteria occupied a wide range of habitats. Quantitative polymerase chain reaction estimated that the abundance of M . oxyfera-like bacteria ranged from 2.2 × 103 to 2.3 × 107 copies g−1 dry soil, and up to around 0.62% of the total number of bacteria. Moreover, the M . oxyfera-like bacteria showed high biodiversity in wetland ecosystems based on the analysis of 462 pmoA and 287 16S rRNA gene sequences. The current study revealed the widespread distribution and biogeography of M . oxyfera-like bacteria in the terrestrial system. [ABSTRACT FROM AUTHOR]

• Nearly complete dissolved methane removal was achieved by integrating PN/anammox/n-DAMO in MABR. • High-level (> 90 %) N removal from mainstream was achieved simultaneously. • N-DAMO microorganisms were the major contributors to dissolved methane removal. • The presence of n-DAMO microorganisms could relax the requirement of NOB suppression. • The system was robust against dynamic influent compositions. Anaerobic technologies with downstream autotrophic nitrogen removal have been proposed to enhance bioenergy recovery and transform a wastewater treatment plant from an energy consumer to an energy exporter. However, approximately 20–50 % of the produced methane is dissolved in the anaerobically treated effluent and is easily stripped into the atmosphere in the downstream aerobic process, contributing to the release of greenhouse gas emissions. This study aims to develop a solution to beneficially utilize dissolved methane to support high-level nitrogen removal from anaerobically treated mainstream wastewater. A novel technology, integrating Partial Nitritation, Anammox and Methane-dependent nitrite/nitrate reduction (i.e. PNAM) was demonstrated in a membrane-aerated biofilm reactor (MABR). With the feeding of ∼50 mg NH 4 +-N/L and ∼20 mg/L dissolved methane at a hydraulic retention time of 15 h, around 90 % of nitrogen and ∼100 % of dissolved methane can be removed together in the MABR. Microbial community characterization revealed that ammonia-oxidizing bacteria (AOB), nitrite-oxidizing bacteria (NOB), anammox bacteria, nitrite/nitrate-dependent anaerobic methane oxidation microorganisms (n-DAMO bacteria and archaea) and aerobic methanotrophs co-existed in the established biofilm. Batch tests confirmed the active microbial pathways and showed that AOB, anammox bacteria and n-DAMO microbes were jointly responsible for the nitrogen removal, and dissolved methane was mainly removed by the n-DAMO process, with aerobic methane oxidation making a minor contribution. In addition, the established system was robust against dynamic changes in influent composition. The study provides a promising technology for the simultaneous removal of dissolved methane and nitrogen from domestic wastewater, which can support the transformation of wastewater treatment from an energy- and carbon-intensive process, to one that is energy- and carbon-neutral. [Display omitted] [ABSTRACT FROM AUTHOR]

• Granules coupling anammox and n-DAMO are used to remove nitrogen and dissolved methane from anaerobic effluent. • Nitrite is a preferred electron acceptor to remove ammonium and dissolved methane. • The low methane affinity of n-DAMO archaea limits methane removal using nitrate as electron acceptor. • Kinetics of anammox bacteria, n-DAMO bacteria, and n-DAMO archaea in granules are revealed and compared. • Microbial cooperation makes the system robust against wastewater dynamics. Anaerobic treatment of mainstream wastewater has been proposed as a promising solution to enhance bioenergy recovery for wastewater treatment plants (WWTPs). However, the limited organics for downstream nitrogen removal and emissions of dissolved methane into the atmosphere are two major barriers to the broad application of anaerobic wastewater treatment. This study aims to develop a novel technology to overcome these two challenges by achieving simultaneous removal of dissolved methane and nitrogen, and unravel the microbial competitions underpinning the process from the microbial and kinetic perspectives. To this end, a laboratory granule-based sequencing batch reactor (GSBR) coupling anammox and nitrite/nitrate-dependent anaerobic methane oxidation (n-DAMO) microorganisms was developed to treat wastewater mimicking effluent from mainstream anaerobic treatment. The GSBR achieved high-level nitrogen and dissolved methane removal rates (> 250 mg N/L/d and > 65 mg CH 4 /L/d) and efficiencies (> 99% total nitrogen removal and > 90% total methane removal) during the long-term demonstration. The availability of different electron acceptors (nitrite or nitrate) imposed significant effects on the removal of ammonium and dissolved methane, as well as on the microbial communities, and the abundance and expression of functional genes. The analysis of apparent microbial kinetics showed that anammox bacteria had a higher nitrite affinity than n-DAMO bacteria, while n-DAMO bacteria had a higher methane affinity than n-DAMO archaea. These kinetics underpin the observation that nitrite is a preferred electron acceptor for removing ammonium and dissolved methane than nitrate. The findings not only extend the applications of novel n-DAMO microorganisms in nitrogen and dissolved methane removal, but also provide insights into microbial cooperation and competition in granular systems. [Display omitted] [ABSTRACT FROM AUTHOR]

• High-level N removal from mainstream was achieved by integrating PN/anammox/n-DAMO. • AOB, anammox and n-DAMO organisms are coupled in a single biofilm. • NOB are suppressed by low DO and nitrite competitors anammox and n-DAMO bacteria. • An average TN removal efficiency above 95% was achieved at a rate of 0.1 kg N/m3/d. • Methane supply is important to remove nitrate in effluent. The application of partial nitritation and anammox to remove nitrogen from mainstream wastewater is of great interest because of the potential to reduce energy cost and carbon dosage. However, this process confronts a dilemma of relatively high effluent nitrogen concentration (>10 mg N/L), owning to the unwanted prevalence of nitrite-oxidizing bacteria (NOB) and the intrinsic nitrate production by anammox bacteria. Here, a novel technology, named the one-stage PNAM, that integrates P artial N itritation, A nammox and M ethane-dependent nitrite/nitrate reduction reactions, was developed in a single membrane biofilm reactor (MBfR). With feeding of synthetic mainstream wastewater containing ∼50 mg NH 4 +-N/L at a hydraulic retention time of 12 h, more than 95% nitrogen was removed in the established one-stage PNAM process at a practically useful rate of 0.1 kg N/m3/d. Microbial community characterization and in-situ batch tests revealed a sophisticated microbial structure consisting of ammonia-oxidizing bacteria (AOB), anammox bacteria, nitrite/nitrate-dependent anaerobic methane oxidation (n-DAMO) bacteria and archaea, and a small fraction of NOB and aerobic methanotrophs. The role of methane in removing nitrate was confirmed by switching on/off the methane supply, which relaxed the requirement for NOB suppression. In addition, the established system was relatively robust against temperature variations, evidenced by a total nitrogen removal efficiency above 80% at temperature as low as 14 ℃. The results provide a promising alternative for efficient nitrogen removal from domestic wastewater using methane as the sole carbon source. [Display omitted] [ABSTRACT FROM AUTHOR]

• Methane-based MBfR was applied to remove nitrate from synthetic groundwater. • The reactor achieved a nitrate removal rate of up to 706 mg-N L−1 d−1. • SCFAs were detected as products of methane bioconversion. • SCFAs were utilized as carbon sources for efficient denitrification. Nitrate (NO 3 −) removal from groundwater is of great importance since it has posed a major risk to human health. Although methane-based denitrification processes have been developed, the practical application of methane as a carbon source for denitrification is constrained by either a long start-up time or a relatively low nitrate removal rate. This study investigated nitrate removal capacity by using a methane-based membrane biofilm reactor (MBfR) feeding with synthetic groundwater. A high nitrate removal rate of 706 mg-N L−1 d−1 was achieved, which is by far the highest rate reported for methane-based denitrification reactors. Short-chain fatty acids (SCFAs) were detected as products of methane bioconversion, with concentrations of acetic and propionic acid up to 168.8 mg L−1 and 50.2 mg L−1, respectively. Long-term operation and batch test results revealed that the SCFAs generated from methane bioconversion were in situ utilized for supporting nitrate removal as carbon and electron sources, thus leading to a practically useful removal rate. The findings suggest that it is promising to apply the methane-based MBfR to remove nitrate from nitrate contaminated groundwater. [ABSTRACT FROM AUTHOR]

It has been reported that microbial reduction of sulfate, nitrite/nitrate and iron/manganese could be coupled with anaerobic oxidation of methane (AOM), which plays a significant role in controlling methane emission from anoxic niches. However, little is known about microbial chromate (Cr(VI)) reduction coupling with AOM. In this study, a microbial consortium was enriched via switching nitrate dosing to chromate feeding as the sole electron acceptor under anaerobic condition in a membrane biofilm reactor (MBfR), in which methane was continuously provided as the electron donor through bubble-less hollow fiber membranes. According to long-term reactor operation and chromium speciation analysis, soluble chromate could be reduced into Cr(III) compounds by using methane as electron donor. Fluorescence in situ hybridization and high-throughput 16S rRNA gene amplicon profiling further indicated that after feeding chromate Candidatus 'Methanoperedens' (a known nitrate-dependent anaerobic methane oxidation archaeon) became sole anaerobic methanotroph in the biofilm, potentially responsible for the chromate bio-reduction driven by methane. Two potential pathways of the microbial AOM-coupled chromate reduction were proposed: (i) Candidatus 'Methanoperedens' independently utilizes chromate as electron acceptor to form Cr(III) compounds, or (ii) Candidatus 'Methanoperedens' oxidizes methane to generate intermediates or electrons, which will be utilized to reduce chromate to Cr(III) compounds by unknown chromate reducers synergistically. Our findings suggest a possible link between the biogeochemical chromium and methane cycles. Unlabelled Image • Microbial Cr(VI) reduction was achieved in a CH4-based MBfR. • Soluble Cr(VI) was reduced to Cr(III) compounds. • DAMO archaea became the sole anaerobic methanotroph in Cr(VI)-reducing biofilm. • DAMO archaea might be responsible for methane-driven Cr(VI) bio-reduction. [ABSTRACT FROM AUTHOR]