Evaluation of hydrolysis and fermentation rates in microbial fuel cells

This study determined the influence of substrate degradation on power generation in microbial fuel cells (MFCs) and microbial community selection on the anode. Air cathode MFCs were fed synthetic medium containing different substrates (acetate, glucose and starch) using primary clarifier sewage as source of electroactive bacteria. The complexity of the substrate affected the MFC performance both for power generation and COD removal. Power output decreased with an increase in substrate complexity from 99 ± 2 mW m⁻² for acetate to 4 ± 2 mW m⁻² for starch. The organic matter removal and coulombic efficiency (CE) of MFCs with acetate and glucose (82% of COD removal and 26% CE) were greater than MFCs using starch (60% of COD removal and 19% of CE). The combined hydrolysis–fermentation rate obtained (0.0024 h⁻¹) was considerably lower than the fermentation rate (0.018 h⁻¹), indicating that hydrolysis of complex compounds limits current output over fermentation. Statistical analysis of microbial community fingerprints, developed on the anode, showed that microbial communities were enriched according to the type of substrate used. Microbial communities producing high power outputs (fed acetate) clustered separately from bacterial communities producing low power outputs (fed complex compounds).     

Anode microbial communities produced by changing from microbial fuel cell to microbial electrolysis cell operation using two different wastewaters

Conditions in microbial fuel cells (MFCs) differ from those in microbial electrolysis cells (MECs) due to the intrusion of oxygen through the cathode and the release of H₂ gas into solution. Based on 16S rRNA gene clone libraries, anode communities in reactors fed acetic acid decreased in species richness and diversity, and increased in numbers of Geobacter sulfurreducens, when reactors were shifted from MFCs to MECs. With a complex source of organic matter (potato wastewater), the proportion of Geobacteraceae remained constant when MFCs were converted into MECs, but the percentage of clones belonging to G. sulfurreducens decreased and the percentage of G. metallireducens clones increased. A dairy manure wastewater-fed MFC produced little power, and had more diverse microbial communities, but did not generate current in an MEC. These results show changes in Geobacter species in response to the MEC environment and that higher species diversity is not correlated with current.     

Energy from algae using microbial fuel cells

Bioelectricity production from a phytoplankton, Chlorella vulgaris, and a macrophyte, Ulva lactuca was examined in single chamber microbial fuel cells (MFCs). MFCs were fed with the two algae (as powders), obtaining differences in energy recovery, degradation efficiency, and power densities. C. vulgaris produced more energy generation per substrate mass (2.5 kWh/kg), but U. lactuca was degraded more completely over a batch cycle (73 ± 1% COD). Maximum power densities obtained using either single cycle or multiple cycle methods were 0.98 W/m² (277 W/m³) using C. vulgaris, and 0.76 W/m² (215 W/m³) using U. lactuca. Polarization curves obtained using a common method of linear sweep voltammetry (LSV) overestimated maximum power densities at a scan rate of 1 mV/s. At 0.1 mV/s, however, the LSV polarization data was in better agreement with single‐ and multiple‐cycle polarization curves. The fingerprints of microbial communities developed in reactors had only 11% similarity to inocula and clustered according to the type of bioprocess used. These results demonstrate that algae can in principle, be used as a renewable source of electricity production in MFCs. Biotechnol. Bioeng. 2009;103: 1068–1076. © 2009 Wiley Periodicals, Inc.     

Microbial community differences between propionate-fed microbial fuel cell systems under open and closed circuit conditions

We report the electrochemical characterization and microbial community analysis of closed circuit microbial fuel cells (CC-MFCs) and open circuit (OC) cells continuously fed with propionate as substrate. Differences in power output between MFCs correlated with their polarization behavior, which is related to the maturation of the anodophilic communities. The microbial communities residing in the biofilm growing on the electrode, biofouled cation-exchange membrane and anodic chamber liquor of OC-and CC-MFCs were characterized by restriction fragment length polymorphism screening of 16S rRNA gene clone libraries. The results show that the CC-MFC anode was enriched in several microorganisms related to known electrochemically active and dissimilatory Fe(III) reducing bacteria, mostly from the Geobacter spp., to the detriment of Bacteroidetes abundant in the OC-MFC anode. The results also evidenced the lack of a specific pelagic community in the liquor sample. The biofilm growing on the cation-exchange membrane of the CC-MFC was found to be composed of a low-diversity community dominated by two microaerophilic species of the Achromobacter and Azovibrio genus.     

Effect of different acclimation methods on the performance of microbial fuel cells using phenol as substrate

Single-chamber microbial fuel cells (MFCs) with air-cathode were constructed. MFCs were fed different feedstocks during their inoculation, their role on phenol degradation and MFC performance were investigated. The results showed that the MFC inoculated using glucose exhibited the highest power density (31.3 mW m^?2) when phenol was used as the sole substrate for MFC. The corresponding biodegradation kinetic constant was obtained at 0.035 h^?1, at an initial phenol concentration of 600 mg L^?1. Moreover, the phenol degradation rates in this MFC with closed circuit were 9.8–16.5 % higher than those in MFC with opened circuit. The cyclic voltammograms revealed a different electrochemical activity of the anode biofilms in the MFC, and this led to differences in performance of the MFCs with phenol as sole substrate. These results demonstrated that phenol degradation and power production are affected by current generation and type of acclimation.     

Change in microbial communities in acetate- and glucose-fed microbial fuel cells in the presence of light

Power densities produced by microbial fuel cells (MFCs) in natural systems are changed by exposure to light through the enrichment of photosynthetic microorganisms. When MFCs with brush anodes were exposed to light (4000 lx), power densities increased by 8–10% for glucose-fed reactors, and 34% for acetate-fed reactors. Denaturing gradient gel electrophoresis (DGGE) profiles based on the 16S rRNA gene showed that exposure to high light levels changed the microbial communities on the anodes. Based on 16S rRNA gene clone libraries of light-exposed systems the anode communities using glucose were also significantly different than those fed acetate. Dominant bacteria that are known exoelectrogens were identified in the anode biofilm, including a purple nonsulfur (PNS) photosynthetic bacterium, Rhodopseudomonas palustris, and a dissimilatory iron-reducing bacterium, Geobacter sulfurreducens. Pure culture tests confirmed that PNS photosynthetic bacteria increased power production when exposed to high light intensities (4000 lx). These results demonstrate that power production and community composition are affected by light conditions as well as electron donors in single-chamber air-cathode MFCs.     

The effect of flavin electron shuttles in microbial fuel cells current production

The effect of electron shuttles on electron transfer to microbial fuel cell (MFC) anodes was studied in systems where direct contact with the anode was precluded. MFCs were inoculated with Shewanella cells, and flavins used as the electron shuttling compound. In MFCs with no added electron shuttles, flavin concentrations monitored in the MFCs' bulk liquid increased continuously with FMN as the predominant flavin. The maximum concentrations were 0.6 μM for flavin mononucleotide and 0.2 μM for riboflavin. In MFCs with added flavins, micro-molar concentrations were shown to increase current and power output. The peak current was at least four times higher in MFCs with high concentrations of flavins (4.5–5.5 μM) than in MFCs with low concentrations (0.2–0.6 μM). Although high power outputs (around 150 mW/m²) were achieved in MFCs with high concentrations of flavins, a Clostridium-like bacterium along with other reactor limitations affected overall coulombic efficiencies (CE) obtained, achieving a maximum CE of 13%. Electron shuttle compounds (flavins) permitted bacteria to utilise a remote electron acceptor (anode) that was not accessible to the cells allowing current production until the electron donor (lactate) was consumed.     

Alteration of bacterial communities and organic matter in microbial fuel cells (MFCs) supplied with soil and organic fertilizer

The alteration of the organic matter (OM) and the composition of bacterial community in microbial fuel cells (MFCs) supplied with soil (S) and a composted organic fertilizer (A) was examined at the beginning and at the end of 3 weeks of incubation under current-producing as well as no-current-producing conditions. Denaturing gradient gel electrophoresis revealed a significant alteration of the microbial community structure in MFCs generating electricity as compared with no-current-producing MFCs. The genetic diversity of cultivable bacterial communities was assessed by random amplified polymorphic DNA (RAPD) analysis of 106 bacterial isolates obtained by using both generic and elective media. Sequencing of the 16S rRNA genes of the more representative RAPD groups indicated that over 50.4% of the isolates from MFCs fed with S were Proteobacteria, 25.1% Firmicutes, and 24.5% Actinobacteria, whereas in MFCs supplied with A 100% of the dominant species belonged to γ-Proteobacteria. The chemical analysis performed by fractioning the OM and using thermal analysis showed that the amount of total organic carbon contained in the soluble phase of the electrochemically active chambers significantly decreased as compared to the no-current-producing systems, whereas the OM of the solid phase became more humified and aromatic along with electricity generation, suggesting a significant stimulation of a humification process of the OM. These findings demonstrated that electroactive bacteria are commonly present in aerobic organic substrates such as soil or a fertilizer and that MFCs could represent a powerful tool for exploring the mineralization and humification processes of the soil OM.     

Biocathode microbial fuel cell for efficient electricity recovery from dairy manure

Microbial fuel cells (MFCs) represent a new biological method for generating electricity directly from biodegradable compounds. Efficiency of MFCs using manure as substrate is generally low. This study proposed a new design by incorporating biocathodes into a three-chamber MFC, which yielded maximum power densities much higher than those reported in literature. The new design placed cylindrical anode chamber for easy stirring and two symmetrical cathodic chambers with reduced anode-cathode distance. The biocathodes were applied to reduce charge transfer resistance. Additionally, biocathode microbial community was cultured to enrich favorable microorganisms. With external loading of 100 Ω, the power densities for new biocathode MFC using 2, 4, 6, 8 and 10% total solids diary manure reached 7.85±1.0 W m(-3), 7.84±1.20 W m(-3), 8.15±0.20 W m(-3), 7.60±0.97 W m(-3) and 5.63±0.97 W m(-3), respectively. The pH drop as a result of manure hydrolysis limited the power output. To provide detailed information of the microbial community in the biocathode MFC, the 454-pyrosequencing technique was adopted. The Firmicutes, γ-, β-, α- and δ-Proteobacteria, Bacteroidetes and Actinobacteria were the major groups on the anode, while γ-, β-, and α-Proteobacteria, Bacteroidetes and Actinobacteria were the predominant groups on the cathode.     

Electrochemical analysis of Clostridium propionicum and its acrylic acid production in microbial fuel cells

Currently, acrylic acid is produced at a low yield by the resting cells of Clostridium propionicum with the supplement of extra electron acceptors. As an alternative way, acrylic acid production coupled with electricity generation was achieved by C. propionicum‐based microbial fuel cells (MFCs). Electricity was generated in the salt‐bridge MFCs with cysteine and resazurin in the anode chamber as mediators, and K₃Fe(CN)₆ as the cathode electron acceptor. Power generation was 21.78 mW/m² with an internal resistance of 9809 Ω. Cyclic voltammograms indicated the main mechanism of power production was the electron transfer facilitated by mediators in the system. In the salt‐bridge MFC system, 0.694 mM acrylic acid was produced together with electricity generation.     

Electricity generation and microbial community changes in microbial fuel cells packed with different anodic materials

Four materials, carbon felt cube (CFC), granular graphite (GG), granular activated carbon (GAC) and granular semicoke (GS) were tested as packed anodic materials to seek a potentially practical material for microbial fuel cells (MFCs). The microbial community and its correlation with the electricity generation performance of MFCs were explored. The maximum power density was found in GAC, followed by CFC, GG and GS. In GAC and CFC packed MFCs, Geobacter was the dominating genus, while Azospira was the most populous group in GG. Results further indicated that GAC was the most favorable for Geobacter adherence and growth, and the maximum power densities had positive correlation with the total biomass and the relative abundance of Geobacter, but without apparent correlation with the microbial diversity. Due to the low content of Geobacter in GS, power generated in this system may be attributed to other microorganisms such as Synergistes, Bacteroidetes and Castellaniella.     

Hydrogen production profiles using furans in microbial electrolysis cells

Microbial electrochemical cells including microbial fuel cells (MFCs) and microbial electrolysis cells (MECs) are novel biotechnological tools that can convert organic substances in wastewater or biomass into electricity or hydrogen. Electroactive microbial biofilms used in this technology have ability to transfer electrons from organic compounds to anodes. Evaluation of biofilm formation on anode is crucial for enhancing our understanding of hydrogen generation in terms of substrate utilization by microorganisms. In this study, furfural and hydroxymethylfurfural (HMF) were analyzed for hydrogen generation using single chamber membrane-free MECs (17 mL), and anode biofilms were also examined. MECs were inoculated with mixed bacterial culture enriched using chloroethane sulphonate. Hydrogen was succesfully produced in the presence of HMF, but not furfural. MECs generated similar current densities (5.9 and 6 mA/cm² furfural and HMF, respectively). Biofilm samples obtained on the 24th and 40th day of cultivation using aromatic compounds were evaluated by using epi-fluorescent microscope. Our results show a correlation between biofilm density and hydrogen generation in single chamber MECs.     

Photosynthetic microbial fuel cells with positive light response

The current study introduces an aerobic single‐chamber photosynthetic microbial fuel cell (PMFC). Evaluation of PMFC performance using naturally growing fresh‐water photosynthetic biofilm revealed a weak positive light response, that is, an increase in cell voltage upon illumination. When the PMFC anodes were coated with electrically conductive polymers, the rate of voltage increased and the amplitude of the light response improved significantly. The rapid immediate positive response to light was consistent with a mechanism postulating that the photosynthetic electron‐transfer chain is the source of the electrons harvested on the anode surface. This mechanism is fundamentally different from the one exploited in previously designed anaerobic microbial fuel cells (MFCs), sediment MFCs, or anaerobic PMFCs, where the electrons are derived from the respiratory electron‐transfer chain. The power densities produced in PMFCs were substantially lower than those that are currently reported for conventional MFC (0.95 mW/m² for polyaniline‐coated and 1.3 mW/m² for polypyrrole‐coated anodes). However, the PMFC did not depend on an organic substrate as an energy source and was powered only by light energy. Its operation was CO₂‐neutral and did not require buffers or exogenous electron transfer shuttles. Biotechnol. Bioeng. 2009; 104: 939–946. © 2009 Wiley Periodicals, Inc.     

Performance and microbial ecology of air-cathode microbial fuel cells with layered electrode assemblies

Microbial fuel cells (MFCs) can be built with layered electrode assemblies, where the anode, proton exchange membrane (PEM), and cathode are pressed into a single unit. We studied the performance and microbial community structure of MFCs with layered assemblies, addressing the effect of materials and oxygen crossover on the community structure. Four MFCs with layered assemblies were constructed using Nafion or Ultrex PEMs and a plain carbon cloth electrode or a cathode with an oxygen-resistant polytetrafluoroethylene diffusion layer. The MFC with Nafion PEM and cathode diffusion layer achieved the highest power density, 381 mW/m² (20 W/m³). The rates of oxygen diffusion from cathode to anode were three times higher in the MFCs with plain cathodes compared to those with diffusion-layer cathodes. Microsensor studies revealed little accumulation of oxygen within the anode cloth. However, the abundance of bacteria known to use oxygen as an electron acceptor, but not known to have exoelectrogenic activity, was greater in MFCs with plain cathodes. The MFCs with diffusion-layer cathodes had high abundance of exoelectrogenic bacteria within the genus Geobacter. This work suggests that cathode materials can significantly influence oxygen crossover and the relative abundance of exoelectrogenic bacteria on the anode, while PEM materials have little influence on anode community structure. Our results show that oxygen crossover can significantly decrease the performance of air-cathode MFCs with layered assemblies, and therefore limiting crossover may be of particular importance for these types of MFCs.     

Pre-acclimation of a wastewater inoculum to cellulose in an aqueous-cathode MEC improves power generation in air-cathode MFCs

Cellulose has been used in two-chamber microbial fuel cells (MFCs), but power densities were low. Higher power densities can be achieved in air-cathode MFCs using an inoculum from a two-chamber, aqueous-cathode microbial electrolysis cell (MEC). Air-cathode MFCs with this inoculum produced maximum power densities of 1070 mW m⁻² (cathode surface area) in single-chamber and 880 mW m⁻² in two-chamber MFCs. Coulombic efficiencies ranged from 25% to 50%, and COD removals were 50-70% based on total cellulose removals of 60-80%. Decreasing the reactor volume from 26 to 14 mL (while maintaining constant electrode spacing) decreased power output by 66% (from 526 to 180 mW m⁻²) due to a reduction in total mass of cellulose added. These results demonstrate that air-cathode MFCs can produce high power densities with cellulose following proper acclimation of the inoculum, and that organic loading rates are important for maximizing power densities from particulate substrates.     

Dynamic changes in the microbial community composition in microbial fuel cells fed with sucrose

The performance and dynamics of the bacterial communities in the biofilm and suspended culture in the anode chamber of sucrose-fed microbial fuel cells (MFCs) were studied by using denaturing gradient gel electrophoresis (DGGE) of PCR-amplified partial 16S rRNA genes followed by species identification by sequencing. The power density of MFCs was correlated to the relative proportions of species obtained from DGGE analysis in order to detect bacterial species or taxonomic classes with important functional role in electricity production. Although replicate MFCs showed similarity in performance, cluster analysis of DGGE profiles revealed differences in the evolution of bacterial communities between replicate MFCs. No correlation was found between the proportion trends of specific species and the enhancement of power output. However, in all MFCs, putative exoelectrogenic denitrifiers and sulphate-reducers accounted for approximately 24% of the bacterial biofilm community at the end of the study. Pareto–Lorenz evenness distribution curves extracted from the DGGE patterns obtained from time course samples indicated community structures where shifts between functionally similar species occur, as observed within the predominant fermentative bacteria. These results suggest the presence of functional redundancy within the anodic communities, a probable indication that stable MFC performance can be maintained in changing environmental conditions. The capability of bacteria to adapt to electricity generation might be present among a wide range of bacteria.     

Effects of the Pt loading side and cathode-biofilm on the performance of a membrane-less and single-chamber microbial fuel cell

In order to analyze the effect of cathode's Pt loading side on the performance of single-chamber microbial fuel cells (MFCs), power generation of a bamboo charcoal membrane-less air-cathode MFC was examined. The maximum power outputs obtained were 0.144 and 1.16 mW, while the maximum voltage outputs were 0.400 and 0.500 V (external resistance was 500 Omega), respectively, when the Pt loading side facing to the air and to the anode chamber solution; after a long time of operation with the side of cathode loaded Pt facing to anode chamber solution, a biofilm was developed on the inner side of cathode. With the formation of this biofilm, the power outputs of MFC increased first, and then decreased to 0.8 mW; oxidation-reduction potentials (ORP) dropped first, and then achieved the level of stability. Coulombic efficiency (CE) increased at a certain extent. In addition, the impact of cathode-biofilm on the loss of water in anode chamber solution was determined.     

Electricity generation from cellulose by rumen microorganisms in microbial fuel cells

In microbial fuel cells (MFCs) bacteria generate electricity by mediating the oxidation of organic compounds and transferring the resulting electrons to an anode electrode. The objective of this study was to test the possibility of generating electricity with rumen microorganisms as biocatalysts and cellulose as the electron donor in two-compartment MFCs. The anode and cathode chambers were separated by a proton exchange membrane and graphite plates were used as electrodes. The medium in the anode chamber was inoculated with rumen microorganisms, and the catholyte in the cathode compartment was ferricyanide solution. Maximum power density reached 55 mW/m(2) (1.5 mA, 313 mV) with cellulose as the electron donor. Cellulose hydrolysis and electrode reduction were shown to support the production of current. The electrical current was sustained for over 2 months with periodic cellulose addition. Clarified rumen fluid and a soluble carbohydrate mixture, serving as the electron donors, could also sustain power output. Denaturing gradient gel electrophoresis (DGGE) of PCR amplified 16S rRNA genes revealed that the microbial communities differed when different substrates were used in the MFCs. The anode-attached and the suspended consortia were shown to be different within the same MFC. Cloning and sequencing analysis of 16S rRNA genes indicated that the most predominant bacteria in the anode-attached consortia were related to Clostridium spp., while Comamonas spp. abounded in the suspended consortia. The results demonstrated that electricity can be generated from cellulose by exploiting rumen microorganisms as biocatalysts, but both technical and biological optimization is needed to maximize power output.     

Bioelectricity production from acidic food waste leachate using microbial fuel cells: Effect of microbial inocula

The effects of three different inocula (domestic wastewater, activated sludge, and anaerobic sludge) on the treatment of acidic food waste leachate in microbial fuel cells (MFCs) were evaluated. A food waste leachate (pH 4.76; 1000 mg chemical oxygen demand (COD)/L) was used as the substrate. The results indicate that the leachate itself can enable electricity production in an MFC, but the co-addition of different inocula significantly reduces the start-up time (approximately 7 days). High COD and volatile fatty acids removal (>87%) were obtained in all MFCs but with only low coulombic efficiencies (CEs) (14–20%). The highest power (432 mW/m³) and CE (20%) were obtained with anaerobic sludge as the co-inoculum. Microbial community analysis (PCR-DGGE) of the established biofilms suggested that the superior performance of the anaerobic sludge-MFC was associated with the enrichment of both fermentative (Clostridium sp. and Bacteroides sp.) and electrogenic bacteria (Magnetospirillum sp. and Geobacter sp.) at the anode.     

Substrate degradation kinetics, microbial diversity, and current efficiency of microbial fuel cells supplied with marine plankton

The decomposition of marine plankton in two-chamber, seawater-filled microbial fuel cells (MFCs) has been investigated and related to resulting chemical changes, electrode potentials, current efficiencies, and microbial diversity. Six experiments were run at various discharge potentials, and a seventh served as an open-circuit control. The plankton consisted of a mixture of freshly captured phytoplankton and zooplankton (0.21 to 1 mm) added at an initial batch concentration of 27.5 mmol liter(-1) particulate organic carbon (OC). After 56.7 days, between 19.6 and 22.2% of the initial OC remained, sulfate reduction coupled to OC oxidation accounted for the majority of the OC that was degraded, and current efficiencies (of the active MFCs) were between 11.3 and 15.5%. In the open-circuit control cell, anaerobic plankton decomposition (as quantified by the decrease in total OC) could be modeled by three terms: two first-order reaction rate expressions (0.79 day(-1) and 0.037 day(-1), at 15 degrees C) and one constant, no-reaction term (representing 10.6% of the initial OC). However, in each active MFC, decomposition rates increased during the third week, lagging just behind periods of peak electricity generation. We interpret these decomposition rate changes to have been due primarily to the metabolic activity of sulfur-reducing microorganisms at the anode, a finding consistent with the electrochemical oxidization of sulfide to elemental sulfur and the elimination of inhibitory effects of dissolved sulfide. Representative phylotypes, found to be associated with anodes, were allied with Delta-, Epsilon-, and Gammaproteobacteria as well as the Flavobacterium-Cytophaga-Bacteroides and Fusobacteria. Based upon these results, we posit that higher current efficiencies can be achieved by optimizing plankton-fed MFCs for direct electron transfer from organic matter to electrodes, including microbial precolonization of high-surface-area electrodes and pulsed flowthrough additions of biomass.