Response of the Anaerobic Methanotrophic Archaeon Candidatus "Methanoperedens nitroreducens" to the Long-Term Ferrihydrite Amendment.

Anaerobic methanotrophic (ANME) archaea can drive anaerobic oxidation of methane (AOM) using solid iron or manganese oxides as the electron acceptors, hypothetically via direct extracellular electron transfer (EET). This study investigated the response of Candidatus "Methanoperedens nitroreducens TS" (type strain), an ANME archaeon previously characterized to perform nitrate-dependent AOM, to an Fe(III)-amended condition over a prolonged period. Simultaneous consumption of methane and production of dissolved Fe(II) were observed for more than 500 days in the presence of Ca. "M. nitroreducens TS," indicating that this archaeon can carry out Fe(III)-dependent AOM for a long period. Ca. "M. nitroreducens TS" possesses multiple multiheme c-type cytochromes (MHCs), suggesting that it may have the capability to reduce Fe(III) via EET. Intriguingly, most of these MHCs are orthologous to those identified in Candidatus "Methanoperedens ferrireducens," an Fe(III)-reducing ANME archaeon. In contrast, the population of Ca. "M. nitroreducens TS" declined and was eventually replaced by Ca. "M. ferrireducens," implying niche differentiation between these two ANME archaea in the environment.

Biochar-Mediated Anaerobic Oxidation of Methane.

Biochar was recently identified as an effective soil amendment for CH4 capture. Corresponding mechanisms are currently recognized to be from physical properties of biochar, providing a favorable growth environment for aerobic methanotrophs which perform aerobic methane (CH4) oxidation. However, our study shows that the chemical reactivity of biochar can also stimulate anaerobic oxidation of CH4 (AOM) by anaerobic methanotrophic archaea (ANME) of ANME-2d, which proposes another plausible mechanism for CH4 mitigation by biochar amendment in anaerobic environments. It was found that, by adding biochar as the sole electron acceptor in an anaerobic environment, CH4 was biologically oxidized, with CO2 production of 106.3 ± 5.1 µmol g-1 biochar. In contrast, limited CO2 production was observed with chemically reduced biochar amendment. This biological nature of the process was confirmed by mcr gene transcript abundance as well as sustained dominance of ANME-2d in the microbial community during microbial incubations with active biochar amendment. Combined FTIR and XPS analyses demonstrated that the redox activity of biochar is related to its oxygen-based functional groups. On the basis of microbial community evolution as well as intermediate production during incubation, different pathways in terms of direct or indirect interactions between ANME-2d and biochar were proposed for biochar-mediated AOM.

Effect of methane partial pressure on the performance of a membrane biofilm reactor coupling methane-dependent denitrification and anammox.

Complete nitrogen removal has recently been demonstrated by integrating anaerobic ammonium oxidation (anammox) and denitrifying anaerobic methane oxidation (DAMO) processes. In this work, the effect of methane partial pressure on the performance of a membrane biofilm reactor (MBfR) consisting of DAMO and anammox microorganisms was evaluated. The activities of DAMO archaea and DAMO bacteria in the biofilm increased significantly with increased methane partial pressure, from 367 ±â€¯9 and 58 ±â€¯22 mg-N L-1d-1 to 580 ±â€¯12 and 222 ±â€¯22 mg-N L-1d-1, respectively, while the activity of anammox bacteria only increased slightly, when the methane partial pressure was elevated from 0.24 to 1.39 atm in the short-term batch tests. The results were supported by a long-term (seven weeks) continuous test, when the methane partial pressure was dropped from 1.39 to 0.78 atm. The methane utilization efficiency was always above 96% during both short-term and long-term tests. Taken together, nitrogen removal rate (especially the nitrate reduction rate by DAMO archaea) and methane utilization efficiency could be maintained at high levels in a broad range of methane partial pressure (0.24-1.39 atm in this study). In addition, a previously established DAMO/anammox biofilm model was used to analyze the experimental data. The observed impacts of methane partial pressure on biofilm activity were well explained by the modeling results. These results suggest that methane partial pressure can potentially be used as a manipulated variable to control reaction rates, ultimately to maintain high nitrogen removal efficiency, according to nitrogen loading rate.

Woodchip-sulfur based heterotrophic and autotrophic denitrification (WSHAD) process for nitrate contaminated water remediation.

Nitrate contaminated water can be effectively treated by simultaneous heterotrophic and autotrophic denitrification (HAD). In the present study, woodchips and elemental sulfur were used as co-electron donors for HAD. It was found that ammonium salts could enhance the denitrifying activity of the Thiobacillus bacteria, which utilize the ammonium that is produced by the dissimilatory nitrate reduction to ammonium (DNRA) in the woodchip-sulfur based heterotrophic and autotrophic denitrification (WSHAD) process. The denitrification performance of the WSHAD process (reaction constants range from 0.05485 h(-1) to 0.06637 h(-1)) is better than that of sulfur-based autotrophic denitrification (reaction constants range from 0.01029 h(-1) to 0.01379 h(-1)), and the optimized ratio of woodchips to sulfur is 1:1 (w/w). No sulfate accumulation is observed in the WSHAD process and the alkalinity generated in the heterotrophic denitrification can compensate for alkalinity consumption by the sulfur-based autotrophic denitrification. The symbiotic relationship between the autotrophic and the heterotrophic denitrification processes play a vital role in the mixotrophic environment.

Pyrite-based autotrophic denitrification for remediation of nitrate contaminated groundwater.

In this study, pyrite-based denitrification using untreated pyrite (UP) and acid-pretreated pyrite (AP) was evaluated as an alternative to elemental sulfur based denitrification. Pyrite-based denitrification resulted in a favorable nitrate removal rate constant (0.95 d(-1)), sulfate production of 388.00 mg/L, and a stable pH. The pretreatment of pyrite with acid led to a further increase in the nitrate removal rate constant (1.03 d(-1)) and reduction in initial sulfate concentration (224.25±7.50 mg/L). By analyzing the microbial community structure using Denaturing Gradient Gel Electrophoresis, it was confirmed that Sulfurimonas denitrificans (S. denitrificans) could utilize pyrite as an electron donor. A stable pH was observed over the entire experimental period, indicating that the use of a pH buffer reagent would not be necessary for pyrite-based denitrification. Therefore, pyrite could effectively replace elemental sulfur as an electron donor in autotrophic denitrification for nitrate-contaminated groundwater remediation.

Optimization of C/N and current density in a heterotrophic/biofilm-electrode autotrophic denitrification reactor (HAD-BER).

In this study, central composite design (CCD) and response surface methodology (RSM) were applied to optimize the C/N and current density in a heterotrophic/biofilm-electrode autotrophic denitrification reactor (HAD-BER). Results showed that nitrate could be effectively reduced over a wide range of C/Ns (0.84-1.3535) and current densities (96.8-370.0 mA/m(2)); however, an optimum C/N of 1.13 and optimum current density of 239.6 mA/m(2) were obtained by RSM. Moreover, the HAD-BER performance under the optimum conditions resulted in almost 100% nitrate-N removal efficiency and low nitrite-N and ammonia-N accumulation. Furthermore, under the optimum conditions, H2 generated from water electrolysis matched the CO2 produced by heterotrophic denitrification by stoichiometric calculation. Therefore, CCD and RSM could be used to acquire optimum operational conditions and improve the nitrate removal efficiency and energy consumption in the HAD-BER.

Characteristics of heterotrophic/biofilm-electrode autotrophic denitrification for nitrate removal from groundwater.

A heterotrophic/biofilm-electrode autotrophic denitrification reactor (HAD-BER) was developed to improve denitrification efficiency and reduce the consumption of organic carbon source. Maximum nitrate removal efficiency of 99.9% was gained under the optimum current density of 200 mA/m(2). The number of heterotrophic denitrification bacteria (HDB) 2.0 × 10(5) and hydrogen autotrophic denitrification bacteria (ADB) 2.0 × 10(3) in per milliliter biofilm solution were observed by the most probable number (MPN) culture, demonstrating that HDB and ADB coexist in the HAD-BER. The inorganic carbon source for autotrophic denitrification was supplied by the dissolved carbon dioxide (CO2) evolved from the heterotrophic denitrification process, indicating that there was synergistic interaction between the HDB and ADB, i.e., the organic carbon source used for denitrification could be decreased in the HAD-BER. Therefore, the developed HAD-BER would be a promising approach for nitrate removal from groundwater.