Characterization of bacterial communities in anode microbial fuel cells fed with glucose,propyl alcohol and methanol

Bacterial communities in anode microbial fuel cells (MFC) obtained from anaerobic digester sludge in a municipal wastewater treatment plant (Nanjing, China) were investigated. Glucose, propyl alcohol and methanol were used as sole carbon source in two-chamber MFC. The results showed that a reproducible cycle of power production can be formed in MFC fed with 3 substrates and glucose-fed MFC had the highest peak power density of 1499 ± 33 mW/m³, followed by methanol- (1264 ± 47 mW/m³) and propyl alcohol-fed MFC (1192 ± 36 mW/m³). Firmicutes, Bacteroidetes, Verrucomicrobia, Proteobacteria, Synergistetes and Armatimonadetes were dominant phyla in 3 MFC. Firmicutes was the most dominant phylum in glucose-fed MFC samples and Bacteroidetes prevailed in methanol- and propyl alcohol-fed MFC. These data indicate that propyl alcohol and methanol along with glucose can be used as substrates of MFC.     

Comparison of anode bacterial communities and performance in microbial fuel cells with different electron donors

Microbial fuel cells (MFCs) harness the electrochemical activity of certain microbes for the production of electricity from reduced compounds. Characterizations of MFC anode biofilms have collectively shown very diverse microbial communities, raising ecological questions about competition and community succession within these anode-reducing communities. Three sets of triplicate, two-chamber MFCs inoculated with anaerobic sludge and differing in energy sources (acetate, lactate, and glucose) were operated to explore these questions. Based on 16S rDNA-targeted denaturing gradient gel electrophoresis (DGGE), all anode communities contained sequences closely affiliated with Geobacter sulfurreducens (>99% similarity) and an uncultured bacterium clone in the Bacteroidetes class (99% similarity). Various other Geobacter-like sequences were also enriched in most of the anode biofilms. While the anode communities in replicate reactors for each substrate generally converged to a reproducible community, there were some variations in the relative distribution of these putative anode-reducing Geobacter-like strains. Firmicutes were found only in glucose-fed MFCs, presumably serving the roles of converting complex carbon into simple molecules and scavenging oxygen. The maximum current density in these systems was negatively correlated with internal resistance variations among replicate reactors and, likely, was only minimally affected by anode community differences in these two-chamber MFCs with high internal resistance.     

Effect of external resistance on bacterial diversity and metabolism in cellulose-fed microbial fuel cells

External resistance affects the performance of microbial fuel cells (MFCs) by controlling the flow of electrons from the anode to the cathode. The purpose of this study was to determine the effect of external resistance on bacterial diversity and metabolism in MFCs. Four external resistances (20, 249, 480, and 1000 Ω) were tested by operating parallel MFCs independently at constant circuit loads for 10 weeks. A maximum power density of 66 mW m⁻² was achieved by the 20 Ω MFCs, while the MFCs with 249, 480, and 1000 Ω external resistances produced 57.5, 27, and 47 mW m⁻², respectively. Denaturing gradient gel electrophoresis analysis of partial 16S rRNA genes showed clear differences between the planktonic and anode-attached populations at various external resistances. Concentrations of short chain fatty acids were higher in MFCs with larger circuit loads, suggesting that fermentative metabolism dominated over anaerobic respiration using the anode as the final electron acceptor.     

Electricity-producing bacterial communities in microbial fuel cells

Microbial fuel cells (MFCs) are not yet commercialized but they show great promise as a method of water treatment and as power sources for environmental sensors. The power produced by these systems is currently limited, primarily by high internal (ohmic) resistance. However, improvements in the system architecture will soon result in power generation that is dependent on the capabilities of the microorganisms. The bacterial communities that develop in these systems show great diversity, ranging from primarily delta-Proteobacteria that predominate in sediment MFCs to communities composed of alpha-, beta-, gamma- or delta-Proteobacteria, Firmicutes and uncharacterized clones in other types of MFCs. Much remains to be discovered about the physiology of these bacteria capable of exocellular electron transfer, collectively defined as a community of "exoelectrogens". Here, we review the microbial communities found in MFCs and the prospects for this emerging bioenergy technology.     

Anodic and cathodic microbial communities in single chamber microbial fuel cells

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The anode potential regulates bacterial activity in microbial fuel cells

The anode potential in microbial fuel cells controls both the theoretical energy gain for the microorganisms as the output of electrical energy. We operated three reactors fed with acetate continuously at a poised anode potential of 0 (R ₀), −200 (R ₋₂₀₀) and −400 (R ₋₄₀₀) mV versus Ag/AgCl and investigated the resulting bacterial activity. The anode potential had no influence on the start-up time of the three reactors. During a 31-day period, R ₋₂₀₀ produced 15% more charge compared to R ₀ and R ₋₄₀₀. In addition, R ₋₂₀₀ had the highest maximal power density (up to 199 W m⁻³ total anode compartment during polarization) but the three reactors evolved to the same power density at the end of the experimental period. During polarization, only the current of R ₋₄₀₀ levelled off at an anode potential of −300 mV versus Ag/AgCl. The maximum respiration rate of the bacteria during batch tests was also considerably lower for R ₋₄₀₀. The specific biomass activity however, was the highest for R ₋₄₀₀ (6.93 g chemical oxygen demand g⁻¹ biomass-volatile suspended solids (VSS) d⁻¹ on day 14). This lowered during the course of the experiment due to an increase of the biomass concentration to an average level of 578 ± 106 mg biomass-VSS L⁻¹ graphite granules for the three reactors. This research indicated that an optimal anode potential of −200 mV versus Ag/AgCl exists, regulating the activity and growth of bacteria to sustain an enhanced current and power generation.     

Methanogenesis versus electrogenesis: morphological and phylogenetic comparisons of microbial communities

Two H-type microbial fuel cells were prepared. The anaerobic chambers were inoculated with rice paddy field soil and fed cellulose as an energy source. In one reactor, the anode and cathode were connected with a wire (closed circuit, CC), while they were not connected in the other reactor (open circuit, OC). The OC reactor actively produced methane. In the CC reactor, however, an electric current of 0.2 to 0.3 mA was constantly generated, and methane production was almost completely suppressed. Electron microscopy revealed that rod-shaped cells with long prosthecae-like filaments were specifically enriched in the CC reactor. Comparisons of 16S rRNA gene clone libraries revealed entirely different phylogenetic compositions in the CC and OC communities; phylotypes related to Rhizobiceae, Desulfovibrio, and Ethanoligenens were specifically enriched in the CC community. The results indicate that electrogenesis resulted in the enrichment of distinctive microbial populations and suppressed methanogenesis from cellulose.     

On the dynamic response of the anode in microbial fuel cells

A study of the dynamic response of a microbial fuel cell (MFC) using membrane electrode assemblies (MEAs) designed for air breathing cathode operation is reported. The MFC used four MEAs simultaneously and has a low internal resistance. An increased concentration of glucose produced a non-linear increase in the maximum current reached. The time to reach the maximum current increased with increasing glucose concentrations of 1-7 mM; varying from approximately 2.4 to 4.2h. The rate at which the current density increased with time was the same for all glucose concentrations up to current densities close to the maximum values. The peak power density varied approximately linearly with glucose concentrations from 2 to 77 mW/m(2) (1-7 mM) with a 1 kΩ resistance. The cell response appeared to be linked to a slow process of fuel transport to the bacteria and their metabolic processes. The dynamic response of the anode was analysed in terms of a substrate mass transport model. The application of different current ranges did not significantly change the dynamic response of either the anode community or the MFC polarization characteristics. Thus, it is likely that the bacterial communities that form under MFC operation contain sufficiently "dominant" electro-active species that are capable of producing high power for MFCs.     

Convergent development of anodic bacterial communities in microbial fuel cells

Microbial fuel cells (MFCs) are often inoculated from a single wastewater source. The extent that the inoculum affects community development or power production is unknown. The stable anodic microbial communities in MFCs were examined using three inocula: a wastewater treatment plant sample known to produce consistent power densities, a second wastewater treatment plant sample, and an anaerobic bog sediment. The bog-inoculated MFCs initially produced higher power densities than the wastewater-inoculated MFCs, but after 20 cycles all MFCs on average converged to similar voltages (470±20 mV) and maximum power densities (590±170 mW m⁻²). The power output from replicate bog-inoculated MFCs was not significantly different, but one wastewater-inoculated MFC (UAJA3 (UAJA, University Area Joint Authority Wastewater Treatment Plant)) produced substantially less power. Denaturing gradient gel electrophoresis profiling showed a stable exoelectrogenic biofilm community in all samples after 11 cycles. After 16 cycles the predominance of Geobacter spp. in anode communities was identified using 16S rRNA gene clone libraries (58±10%), fluorescent in-situ hybridization (FISH) (63±6%) and pyrosequencing (81±4%). While the clone library analysis for the underperforming UAJA3 had a significantly lower percentage of Geobacter spp. sequences (36%), suggesting that a predominance of this microbe was needed for convergent power densities, the lower percentage of this species was not verified by FISH or pyrosequencing analyses. These results show that the predominance of Geobacter spp. in acetate-fed systems was consistent with good MFC performance and independent of the inoculum source.     

Microbial fuel cells meet with external resistance

The influence of external load on the composition of the anodic biofilm microbial community and biomass yield was investigated in a microbial fuel cell fed with glucose and domestic wastewater was used as source of electrogens. Denaturing gradient gel electrophoresis (DGGE) of polymerase chain reaction (PCR) amplified 16S rRNA gene fragments revealed distinct differences in anodic bacterial communities formed at the anode of each MFC operated under a different external load. These results implied that in an MFC, electrogenic bacteria were enriched under higher current densities, i.e., low external load, and were able to sustain better current and effluent quality. The influence of the external resistance applied to the MFCs during formation of the bacterial communities from sewage wastewater was shown to have no significant effect on power performance of the MFCs nor to have a significant influence on their anodic activity with both glucose and brewery wastewater as fuel. As expected, current generation, COD removal and the biomass yield were all directly influenced by the external load. Significantly, when operated under lower external load, the biomass yield in the MFC was less than that in conventional anaerobic digestion (i.e., control).     

Impact of initial biofilm growth on the anode impedance of microbial fuel cells

Electrochemical impedance spectroscopy (EIS) was used to study the behavior of a microbial fuel cell (MFC) during initial biofilm growth in an acetate-fed, two-chamber MFC system with ferricyanide in the cathode. EIS experiments were performed both on the full cell (between cathode and anode) as well as on individual electrodes. The Nyquist plots of the EIS data were fitted with an equivalent electrical circuit to estimate the contributions of various intrinsic resistances to the overall internal MFC impedance. During initial development of the anode biofilm, the anode polarization resistance was found to decrease by over 70% at open circuit and by over 45% at 27 microA/cm(2), and a simultaneous increase in power density by about 120% was observed. The exchange current density for the bio-electrochemical reaction on the anode was estimated to be in the range of 40-60 nA/cm(2) for an immature biofilm after 5 days of closed circuit operation, which increased to around 182 nA/cm(2) after more than 3 weeks of operation and stable performance in an identical parallel system. The polarization resistance of the anode was 30-40 times higher than that of the ferricyanide cathode for the conditions tested, even with an established biofilm. For a two-chamber MFC system with a Nafion 117 membrane and an inter-electrode spacing of 15 cm, the membrane and electrolyte solution dominate the ohmic resistance and contribute to over 95% of the MFC internal impedance. Detailed EIS analyses provide new insights into the dominant kinetic resistance of the anode bio-electrochemical reaction and its influence on the overall power output of the MFC system, even in the high internal resistance system used in this study. These results suggest that new strategies to address this kinetic constraint of the anode bio-electrochemical reactions are needed to complement the reduction of ohmic resistance in modern designs.     

Power production enhancement with a polyaniline modified anode in microbial fuel cells

In this paper, an approach of improving power generation of microbial fuel cells (MFCs) by using a HSO(4)(-) doped polyaniline modified carbon cloth anode was reported. The modification of carbon cloth anode was accomplished by electrochemical polymerization of aniline in 5% H(2)SO(4) solution. A dual-chamber MFC reactor with the modified anode achieved a maximum power density of 5.16 Wm(-3), an internal resistance of 90 Ω, and a start-up time of 4 days, which was respectively 2.66 times higher, 65.5% lower, and 33.3% shorter than the corresponding values of the MFC with unmodified anode. Evidence from X-ray photoelectron spectroscopy and scanning electron microscopy results proved that the formation of biofilm on the anode surface could prevent the HSO(4)(-) doped polyaniline to be de-doped, and the results from electrochemical tests confirmed that the electrochemical activity of the modified anode was enhanced significantly after inoculation. Charge transfer was facilitated by polyaniline modification. All the results indicated that the polyaniline modification on the anode was an efficient approach of improving the performance of MFCs.     

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.     

Microorganism-immobilized carbon nanoparticle anode for microbial fuel cells based on direct electron transfer

A fast and convenient bacterial immobilization method was proposed as an attempt to improve the anode efficiency of a microbial fuel cell, in which bacteria were entrapped into carbon nanoparticle matrix. The direct electron transfer from the entrapped bacterial cells to the anode was verified using cyclic voltammogram (CV). Using the immobilized bioanode, the start-up time of the MFC was greatly reduced. Meanwhile, the maximum power density of 1,947 mW m⁻² with the modified anode was much higher than that with the biofilm-based carbon cloth anode (1,479 mW m⁻²). Impedance measurements suggested that performance improvement resulted from the decrease in charge transfer and diffusion resistances. The results demonstrated that bacteria immobilization using carbon nanoparticle matrix was a simple and efficient approach for improving the anodes performances in MFCs.     

Loading rate and external resistance control the electricity generation of microbial fuel cells with different three-dimensional anodes

The electricity generation, electrochemical and microbial characteristics of five microbial fuel cells (MFCs) with different three-dimensional electrodes (graphite and carbon felt, 2mm and 5mm graphite granules and graphite wool) was examined in relation to the applied loading rate and the external resistance. The graphite felt electrode yielded the highest maximum power output amounting up to 386Wm(-3) total anode compartment (TAC). However, based on the continuous current generation, limited differences between the materials were registered. Doubling the loading rate to 3.3gCODL(-1)TACd(-1) resulted only in an increased current generation when the external resistance was low (10.5-25Omega) or during polarization. Conversely, lowering the external resistance resulted in a steady increase of both the kinetic capacities of the biocatalyst and the continuous current generation from 77 (50Omega) up to 253 (10.5Omega)Am(-3)TAC. Operating a MFC at an external resistance close to its internal resistance, allows to increase the current generation from enhanced loading rates while maximizing the power generation.     

Shifting microbial communities sustain multiyear iron reduction and methanogenesis in ferruginous sediment incubations

Reactive Fe(III) minerals can influence methane (CH₄) emissions by inhibiting microbial methanogenesis or by stimulating anaerobic CH₄ oxidation. The balance between Fe(III) reduction, methanogenesis, and CH₄ oxidation in ferruginous Archean and Paleoproterozoic oceans would have controlled CH₄ fluxes to the atmosphere, thereby regulating the capacity for CH₄ to warm the early Earth under the Faint Young Sun. We studied CH₄ and Fe cycling in anoxic incubations of ferruginous sediment from the ancient ocean analogue Lake Matano, Indonesia, over three successive transfers (500 days in total). Iron reduction, methanogenesis, CH₄ oxidation, and microbial taxonomy were monitored in treatments amended with ferrihydrite or goethite. After three dilutions, Fe(III) reduction persisted only in bottles with ferrihydrite. Enhanced CH₄ production was observed in the presence of goethite, highlighting the potential for reactive Fe(III) oxides to inhibit methanogenesis. Supplementing the media with hydrogen, nickel and selenium did not stimulate methanogenesis. There was limited evidence for Fe(III)‐dependent CH₄ oxidation, although some incubations displayed CH₄‐stimulated Fe(III) reduction. 16S rRNA profiles continuously changed over the course of enrichment, with ultimate dominance of unclassified members of the order Desulfuromonadales in all treatments. Microbial diversity decreased markedly over the course of incubation, with subtle differences between ferrihydrite and goethite amendments. These results suggest that Fe(III) oxide mineralogy and availability of electron donors could have led to spatial separation of Fe(III)‐reducing and methanogenic microbial communities in ferruginous marine sediments, potentially explaining the persistence of CH₄ as a greenhouse gas throughout the first half of Earth history.     

Long-term cathode performance and the microbial communities that develop in microbial fuel cells fed different fermentation endproducts

To better understand how cathode performance and substrates affected communities that evolved in these reactors over long periods of time, microbial fuel cells were operated for more than 1 year with individual endproducts of lignocellulose fermentation (acetic acid, formic acid, lactic acid, succinic acid, or ethanol). Large variations in reactor performance were primarily due to the specific substrates, with power densities ranging from 835 ± 21 to 62 ± 1 mW/m³. Cathodes performance degraded over time, as shown by an increase in power of up to 26% when the cathode biofilm was removed, and 118% using new cathodes. Communities that developed on the anodes included exoelectrogenic families, such as Rhodobacteraceae, Geobacteraceae, and Peptococcaceae, with the Deltaproteobacteria dominating most reactors. Pelobacter propionicus was the predominant member in reactors fed acetic acid, and it was abundant in several other MFCs. These results provide valuable insights into the effects of long-term MFC operation on reactor performance.