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Microbial electrosynthesis is a useful form of technology for the renewable production of organic commodities from biologically catalyzed reduction of CO₂. However, for the technology to become applicable, process selectivity, stability and efficiency need strong improvement. Here we report on the effect of different electrochemical control modes (potentiostatic/galvanostatic) on both the start-up characteristics and steady-state performance of biocathodes using a non-enriched mixed-culture inoculum. Based on our results, it seems that kinetic differences exist between the two dominant functional microbial groups (i.e., homoacetogens and methanogens) and that by applying different current densities, these differences may be exploited to steer product selectivity and reactor performance., Competing Interests: The authors declare no conflict of interest.

Bioelectrochemical power-to-gas (BEP2G) is considered a potentially convenient way of storing renewable surplus electricity in the form of methane. In methane-producing bioelectrochemical systems (BESs), carbon dioxide and electrical energy are converted into methane, using electrodes that supply either electrons or hydrogen to methanogenic archaea. This review summarizes the performance of methane-producing BESs in relation to cathode potential, electrode materials, operational strategies, and inoculum. Analysis and estimation of energy input and production rates show that BEP2G may become an attractive alternative for thermochemical methanation, and biochemical methanogenesis. To determine if BEP2G can become a future power-to-gas technology, challenges relating to cathodic energy losses, choice of a suitable electron donor, efficient reactor design/operation, and experience with large reactors need to be overcome., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

Although literature suggests that aquatic worms can help to enhance the methane production from excess activated sludge, clear evidence for this is missing. Therefore, anaerobic digestion tests were performed at 20 and at 30°C with sludge from a high-loaded membrane bioreactor, the aquatic worm Lumbriculus variegatus, feces from these worms and with mixtures of these substrates. A significant synergistic effect of the worms or their feces on methane production from the high-loaded sludge or on its digestion rate was not observed. However, a positive effect on low-loaded activated sludge, which generally has a lower anaerobic biodegradability, cannot be excluded. The results furthermore showed that the high-loaded sludge provides an excellent feed for L. variegatus, which is promising for concepts where worm biomass is considered a resource for technical grade products such as coatings and glues., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

The use of methane and acetate as electron donors for biological reduction of thiosulphate in a 5-L laboratory membrane bioreactor was studied and compared to disproportionation of thiosulphate as competing biological reaction. The reactor was operated for 454 days in semi-batch mode; 30 % of its liquid phase was removed and periodically replenished (days 77, 119, 166, 258, 312 and 385). Although the reactor was operated under conditions favourable to promote thiosulphate reduction coupled to methane oxidation, thiosulphate disproportionation was the dominant microbial process. Pyrosequencing analysis showed that the most abundant microorganisms in the bioreactor were phototrophic green sulphur bacteria (GSB) belonging to the family Chlorobiaceae and thiosulphate-disproportionating bacteria belonging to the genus Desulfocapsa. Even though the reactor system was surrounded with opaque plastic capable of filtering most of the light, the GSB used it to oxidize the hydrogen sulphide produced from thiosulphate disproportionation to elemental sulphur. Interrupting methane and acetate supply did not have any effect on the microbial processes taking place. The ultimate goal of our research was to develop a process that could be applied for thiosulphate and sulphate removal and biogenic sulphide formation for metal precipitation. Even though the system achieved in this study did not accomplish the targeted conversion using methane as electron donor, it does perform microbial conversions which allow to directly obtain elemental sulphur from thiosulphate.

Anaerobic methane oxidation coupled to sulphate reduction and the use of ethane and propane as electron donors by sulphate-reducing bacteria represent new opportunities for the treatment of streams contaminated with sulphur oxyanions. However, growth of microbial sulphate-reducing populations with methane, propane or butane is extremely slow, which hampers research and development of bioprocesses based on these conversions. Thermodynamic calculations indicate that the growth rate with possible alternative terminal electron acceptors such as thiosulphate and elemental sulphur may be higher, which would facilitate future research. Here, we investigate the use of these electron acceptors for oxidation of methane, ethane and propane, with marine sediment as inoculum. Mixed marine sediments originating from Aarhus Bay (Denmark) and Eckernförde Bay (Germany) were cultivated anaerobically at a pH between 7.2 and 7.8 and a temperature of 15 °C in the presence of methane, ethane and propane and various sulphur electron acceptors. The sulphide production rates in the conditions with methane, ethane and propane with sulphate were respectively 2.3, 2.2 and 1.8 μmol S L(-1) day(-1). For sulphur, no reduction was demonstrated. For thiosulphate, the sulphide production rates were up to 50 times higher compared to those of sulphate, with 86.2, 90.7 and 108.1 μmol S L(-1) day(-1) for methane, ethane and propane respectively. This sulphide production was partly due to disproportionation, 50 % for ethane but only 7 and 14 % for methane and propane respectively. The oxidation of the alkanes in the presence of thiosulphate was confirmed by carbon dioxide production. This is, to our knowledge, the first report of thiosulphate use as electron acceptor with ethane and propane as electron donors. Additionally, these results indicate that thiosulphate is a promising electron acceptor to increase start-up rates for sulphate-reducing bioprocesses coupled to short-chain alkane oxidation.

Simultaneous nitrogen and methane removal by the slow growing denitrifying methanotrophic bacterium 'Candidatus Methylomirabilis oxyfera' offers opportunities for a new approach to wastewater treatment. However, volumetric nitrite consumption rates should be increased by an order of magnitude before application in wastewater treatment becomes possible. A maximum volumetric nitrite consumption rate of 36 mg NO2(-)-N/L d was achieved in a membrane bioreactor inoculated with wastewater sludge and operated at 20°C. This rate is similar to maximum rates reported in literature, though it was thought that by strict biomass retention using membranes, higher rates would be achieved. In experiments lasting several years, growth was not stable: every experiment showed a decrease in activity after 1-2 years. The cause remains unknown. Rates increased after addition of copper and operating a membrane bioreactor at shorter hydraulic retention times. Further research should focus on long-term effects of copper addition and operation at hydraulic retention times in the order of hours using membrane bioreactors., (Copyright © 2014 Elsevier B.V. All rights reserved.)

A methane-producing biocathode that converts CO(2) into methane was studied electrochemically and microbiologically. The biocathode produced methane at a maximum rate of 5.1 L CH(4)/m(2) projected cathode per day (1.6 A/m(2)) at -0.7 V versus NHE cathode potential and 3.0 L CH(4)/m(2) projected cathode per day (0.9 A/m(2)) at -0.6 V versus NHE cathode potential. The microbial community at the biocathode was dominated by three phylotypes of Archaea and six phylotypes of bacteria. The Archaeal phylotypes were most closely related to Methanobacterium palustre and Methanobacterium aarhusense. Besides methanogenic Archaea, bacteria seemed to be associated with methane production, producing hydrogen as an intermediate. Biomass density varied greatly with part of the carbon electrode covered with a dense biofilm, while only clusters of cells were found on other parts. Based on our results, we discuss how inoculum enrichment and changing operational conditions may help to increase biomass density and to select for microorganisms that produce methane.

This study investigates the oxidation of labeled methane (CH(4)) and the CH(4) dependence of sulfate reduction in three types of anaerobic granular sludge. In all samples, (13)C-labeled CH(4) was anaerobically oxidized to (13)C-labeled CO(2), while net endogenous CH(4) production was observed. Labeled-CH(4) oxidation rates followed CH(4) production rates, and the presence of sulfate hampered both labeled-CH(4) oxidation and methanogenesis. Labeled-CH(4) oxidation was therefore linked to methanogenesis. This process is referred to as trace CH(4) oxidation and has been demonstrated in methanogenic pure cultures. This study shows that the ratio between labeled-CH(4) oxidation and methanogenesis is positively affected by the CH(4) partial pressure and that this ratio is in methanogenic granular sludge more than 40 times higher than that in pure cultures of methanogens. The CH(4) partial pressure also positively affected sulfate reduction and negatively affected methanogenesis: a repression of methanogenesis at elevated CH(4) partial pressures confers an advantage to sulfate reducers that compete with methanogens for common substrates, formed from endogenous material. The oxidation of labeled CH(4) and the CH(4) dependence of sulfate reduction are thus not necessarily evidence of anaerobic oxidation of CH(4) coupled to sulfate reduction.

Anaerobic oxidation of methane (AOM) in marine sediments is coupled to sulfate reduction (SR). AOM is mediated by distinct groups of archaea, called anaerobic methanotrophs (ANME). ANME co-exist with sulfate-reducing bacteria, which are also involved in AOM coupled SR. The microorganisms involved in AOM coupled to SR are extremely difficult to grow in vitro. Here, a novel well-mixed submerged-membrane bioreactor system is used to grow and enrich the microorganisms mediating AOM coupled to SR. Four reactors were inoculated with sediment sampled in the Eckernförde Bay (Baltic Sea) and operated at a methane and sulfate loading rate of 4.8 L L(-1) day(-1) (196 mmol L(-1) day(-1)) and 3.0 mmol L(-1) day(-1). Two bioreactors were controlled at 15 degrees C and two at 30 degrees C, one reactor at 30 degrees C contained also anaerobic granular sludge. At 15 degrees C, the volumetric AOM and SR rates doubled approximately every 3.8 months. After 884 days, an enrichment culture was obtained with an AOM and SR rate of 1.0 mmol g(volatile suspended solids) (-1) day(-1) (286 micromol g(dry weight) (-1) day(-1)). No increase in AOM and SR was observed in the two bioreactors operated at 30 degrees C. The microbial community of one of the 15 degrees C reactors was analyzed. ANME-2a became the dominant archaea. This study showed that sulfate reduction with methane as electron donor is possible in well-mixed bioreactors and that the submerged-membrane bioreactor system is an excellent system to enrich slow-growing microorganisms, like methanotrophic archaea.

Sulfate reduction (SR) coupled to anaerobic oxidation of methane (AOM) is meditated by marine microorganisms and forms an important process in the global sulfur and carbon cycle. In this research, the possibility to use this process for the removal and recovery of sulfur and metal compounds from waste streams was investigated. A membrane bioreactor was used to enrich for a community of methane-oxidizing sulfate-reducing microorganisms from Eckernförde Bay sediment The AOM and SR rate of the obtained enrichment were 1.0 mmol gvss(-1) d(-1). The operational window and optimal environmental conditions for SR with methane as electron donor were assessed. The optimum pH, salinity, and temperature were 7.5, 30% per hundred and 20 degrees C, respectively. The enrichment had a good affinity for sulfate (Km < 0.5 mM) and a low affinity for methane (Kn > 0.075 MPa). A0M coupled to SR was completely inhibited at 2.4 (L0.1) mM sulfide. AOM occurred with sulfate, thiosulfate, and sulfite as electron accepters. Sulfate reduction with methane as electron donor can be applied for the removal of sulfate or for the production of sulfide,for metal precipitation. However, the low optimal temperature and the high salt requirement limit the operational window of the process.

BACKGROUND In a methane-producing bioelectrochemical system ( BES) microorganisms grow on an electrode and catalyse the conversion of CO2 and electricity into methane. Theoretically, methane can be produced bioelectrochemically from CO2 via direct electron transfer or indirectly via hydrogen, acetate or formate. Understanding the electron transfer mechanisms could give insight into methods to steer the process towards higher rate. RESULTS In this study, the electron transfer mechanisms of bioelectrochemical methane production by mixed cultures were investigated. At a cathode potential of −0.7 V vs. normal hydrogen electrode ( NHE), average current density was 2.9 A m−2 cathode and average methane production rate was 1.8 mole e− eq m−2 cathode per day (5.2 L CH4 m−2 cathode per day). Methane was primarily produced indirectly via hydrogen and acetate. Methods to steer towards bioelectrochemical hydrogen and acetate production to further improve the performance of a methane-producing BES are discussed. CONCLUSION At cathode potentials equal to or lower than −0.7 V vs. NHE and using mixed cultures, methane was primarily produced indirectly via hydrogen and acetate. (Bio)electrochemical hydrogen and acetate production rate could be increased by optimizing the cathode design and by enriching the microbial community. Consequently, the production rate of CO2-neutral methane in a BES could be increased. © 2014 Society of Chemical Industry [ABSTRACT FROM AUTHOR]

SUMMARY A methane-producing microbial electrolysis cell (MEC) is a technology to convert CO2 into methane, using electricity as an energy source and microorganisms as the catalyst. A methane-producing MEC provides the possibility to increase the fuel yield per hectare of land area, when the CO2 produced in biofuel production processes is converted to additional fuel methane. Besides increasing fuel yield per hectare of land area, this also results in more efficient use of land area, water, and nutrients. In this research, the performance of a methane-producing MEC was studied for 188 days in a flat-plate MEC design. Methane production rate and energy efficiency of the methane-producing MEC were investigated with time to elucidate the main bottlenecks limiting system performance. When using water as the electron donor at the anode during continuous operation, methane production rate was 0.006 m3/m3 per day at a cathode potential of −0.55 V vs. normal hydrogen electrode with a coulombic efficiency of 23.1%. External electrical energy input was 73.5 kWh/m3 methane, resulting in a voltage efficiency of 13.4%. Consequently, overall energy efficiency was 3.1%. The maximum achieved energy efficiency was obtained in a yield test and was 51.3%. Analysis of internal resistance showed that in the short term, cathode and anode losses were dominant, but with time, also pH gradient and transport losses became more important. The results obtained in this study are used to discuss the possible contribution of methane-producing MECs to increase the fuel yield per hectare of land area. Copyright © 2011 John Wiley & Sons, Ltd. [ABSTRACT FROM AUTHOR]

[Display omitted] • Ca2+ addition doubled the SRT and increased hydrolysis from 29 % to 67 %. • Methanization increased from 20 % to 46 % by adding Ca2+.. • Ca2+ effect on methane production depends on Ca:COD (optimum 0.03 – 0.05, limit 0.08). • Phosphorus removal increased from 38 % to 61 % by adding Ca2+ to the UASB reactor. • Ca2+ was captured by bicarbonate, limiting Ca x (PO 4) y formation and struvite dissolution. Cow manure has potential to serve as a sustainable secondary fuel and phosphorus resource and cut our reliance on finite primary resources. The efficient and sustainable valorization of cow manure faces compositional challenges because of a high solids content, the lack of soluble phosphorus, fine struvite particles being the main phosphorus species and high bicarbonate concentrations. Addition of calcium could result in higher methane production and conversion of struvite to calcium phosphate. To investigate this, cow manure was digested in two up-flow anaerobic sludge blanket reactors for 456 days, one with and one without CaCl 2 addition. A positive effect of calcium addition was found for hydrolysis (29 % without calcium and 67 % with calcium), methane production (136 L-CH 4 kgVS-1 without calcium and 301 L-CH 4 kgVS-1 with calcium) and sludge bed development. Although calcium was added in a 3:1 ratio to phosphorus, it did not result in recrystallization of struvite to calcium phosphate. Instead, it precipitated as calcium carbonate, which was further induced by additional bicarbonate production through higher hydrolysis and methane production. Still, calcium addition caused better phosphorous removal (from 38 % to 61 %), which is attributed to the enhanced sludge bed capturing and accumulating both the calcium carbonate and struvite fines. Higher methane production and improved phosphorus retention enables better valorization of cow manure. The access to resources from cow manure through this technology can contribute to the circularity of agriculture and save on finite natural resources. [ABSTRACT FROM AUTHOR]

Anaerobic digestion at low temperature is an attractive technology especially in moderate climates, however, low temperature results in low microbial activity and low rates of methane formation. This study investigated if bioelectrochemical systems (BESs) can enhance methane production from organic matter in low-temperature anaerobic digestion (AD). A bioelectrochemical reactor was operated with granular activated carbon as electrodes at 10 °C. Our results showed that bioelectrochemical systems can enhance CH 4 yield, accelerate CH 4 production rate and increase acetate removal efficiency at 10 °C. The highest CH 4 yield of 31 mg CH 4 -COD/g VSS was achieved in the combined BES-AD system at a cathode potential of −0.90 V (Ag/AgCl), which was 5.3–6.6 times higher than that in the AD reactor at 10 °C. CH 4 production rate achieved in the combined BES-AD system at 10 °C was only slightly lower than that in the AD reactor at 30 °C. The presence of an external circuit between the acetate-oxidizing bioanode and methane-producing cathode provided an alternative pathway from acetate via electrons to methane, potentially via hydrogen. This alternative pathway seems to result in higher CH 4 production rates at low temperature compared with traditional methanogenesis from acetate. Integration of BES with AD could therefore be an attractive alternative strategy to enhance the performance of anaerobic digestion in cold areas. [ABSTRACT FROM AUTHOR]

Abstract: Despite many advantages of anaerobic sewage treatment over conventional activated sludge treatment, it has not yet been applied in temperate zones. This is especially because effluent from low-temperature anaerobic treatment contains nitrogen and dissolved methane. The presence of nitrogen and methane offers the opportunity to develop a reactor in which methane is used as electron donor for denitrification. Such a reactor could be used in a new concept for low-temperature anaerobic sewage treatment, consisting of a UASB-digester system, a reactor for denitrification coupled to anaerobic methane oxidation, and a nitritation reactor. In the present study denitrifying methanotrophic bacteria similar to ‘Candidatus Methylomirabilis oxyfera’ were enriched. Maximum volumetric nitrite consumption rates were 33.5mgNO2 −-N/Ld (using synthetic medium) and 37.8mgNO2 −-N/Ld (using medium containing effluent from a sewage treatment plant), which are similar to the maximum rate reported so far. Though the goal was to increase the rates, in both reactors, after reaching these maximum rates, volumetric nitrite consumption rates decreased in time. Results indicate biomass washout may have significantly decelerated enrichment. Therefore, to obtain higher volumetric consumption rates, further research should focus on systems with complete biomass retention. [Copyright &y& Elsevier]

Adding calcium during anaerobic digestion of vacuum collected black water (BW) in an up-flow anaerobic sludge bed (UASB) reactor increased the retention of total phosphorus (P) in the reactor from 51% to 87%. However, the insufficient mixing in the reactor caused cementation and relatively high content of organics in the recovered calcium phosphate (CaP) granules, limiting the P recovery. In this study, the UASB reactor was mixed with an internal gas-lift (UASB-GL) to prevent cementation and to enhance the P content in CaP granules. The novel UASB-GL reactor operated for 300 days, treating concentrated BW. At steady state, the removal of total COD and P was 92% and 90%, respectively. The gas injection created a sludge bed with an average total suspended solids concentration of 73 ± 16 g/L at the bottom and 31 ± 5 g/L at the top of the reactor. The concentration of solid P at the bottom of the reactor was 4.58 ± 1.34 gP/L, while at the top a much lower concentration was obtained (0.75 ± 0.32 gP/L). 89% of the CaP granules was found at the bottom of the reactor. The harvested CaP granules (>0.4 mm diameter) contained on average 7.8 ± 0.6 wt% of P. A potential recovery of 57% of P in BW as CaP granules was calculated, considering actual application of the UASB-GL reactor in source separated sanitation. Image 1 • Phosphorus removal was >90% during stable operation at 0.96 gCOD/L ⋅ d. • The phosphorous content in recovered CaP granules increased 16% by gas-lift mixing. • 70% of the phosphorus at the harvesting location was as CaP granules with 7.8 wt% of P. [ABSTRACT FROM AUTHOR]