Effect of formation of biofilms and chemical scale on the cathode electrode on the performance of a continuous two-chamber microbial fuel cell

A two-chamber MFC system was operated continuously for more than 500 days to evaluate effects of biofilm and chemical scale formation on the cathode electrode on power generation. A stable power density of 0.57 W/m² was attained after 200 days operation. However, the power density decreased drastically to 0.2 W/m² after the cathodic biofilm and chemical scale were removed. As the cathodic biofilm and chemical scale partially accumulated on the cathode, the power density gradually recovered with time. Microbial community structure of the cathodic biofilm was analyzed based on 16S rRNA clone libraries. The clones closely related to Xanthomonadaceae bacterium and Xanthomonas sp. in the Gammaproteobacteria subdivision were most frequently retrieved from the cathodic biofilm. Results of the SEM-EDX analysis revealed that the cation species (Na⁺ and Ca²⁺) were main constituents of chemical scale, indicating that these cations diffused from the anode chamber through the Nafion membrane. However, an excess accumulation of the biofilm and chemical scale on the cathode exhibited adverse effects on the power generation due to a decrease in the active cathode surface area and an increase in diffusion resistance for oxygen. Thus, it is important to properly control the formation of chemical scale and biofilm on the cathode during long-term operation.     

Bioengineering of the Enterobacter aerogenes strain for biohydrogen production

Enterobacter aerogenes is one of the most widely-studied model strains for fermentative hydrogen production. To improve the hydrogen yield of E. aerogenes, the bioengineering on a biomolecular level and metabolic network level is of importance. In this review, the fermentative technology of E. aerogenes for hydrogen production will be first briefly summarized. And then the bioengineering of E. aerogenes for the improvement of hydrogen yield will be thoroughly reviewed, including the anaerobic metabolic networks for hydrogen evolution in E. aerogenes, metabolic engineering for improving hydrogen production in E. aerogenes and mixed culture of E. aerogenes with other hydrogen-producing bacteria to enhance the overall yield in anaerobic cultivation. Finally, a perspective on E. aerogenes as a hydrogen producer including systems bioengineering approach for improving the hydrogen yield and application of the engineered E. aerogenes in mixed culture will be presented.     

The utility of Shewanella japonica for microbial fuel cells

Shewanella-containing microbial fuel cells (MFCs) typically use the fresh water wild-type strain Shewanella oneidensis MR-1 due to its metabolic diversity and facultative oxidant tolerance. However, S. oneidensis MR-1 is not capable of metabolizing polysaccharides for extracellular electron transfer. The applicability of Shewanella japonica (an agar-lytic Shewanella strain) for power applications was analyzed using a diverse array of carbon sources for current generation from MFCs, cellular physiological responses at an electrode surface, biofilm formation, and the presence of soluble extracellular mediators for electron transfer to carbon electrodes. Critically, air-exposed S. japonica utilizes biosynthesized extracellular mediators for electron transfer to carbon electrodes with sucrose as the sole carbon source.     

Effects of biomass weight and light intensity on the performance of photosynthetic microbial fuel cells with Spirulina platensis

Microalgae Spirulina platensis were attached to the anode of a membrane-free and mediator-free microbial fuel cell (MFC) to produce electricity through the consumption of biochemical compounds inside the microalgae. An increase in open circuit voltage (OCV) was observed with decreasing light intensity and optimal biomass area density. The highest OCV observation for the MFC was 0.39 V in the dark with a biomass area density on the anode surface of 1.2 g cm⁻². Additionally, it was observed that the MFC with 0.75 g cm⁻² of biomass area density produced 1.64 mW m⁻² of electrical power in the dark, which is superior to the 0.132 mW m⁻² produced in the light. Which also means the MFC can be applied to generate electrical power under both day and night conditions.     

Impact of anodophilic biofilm bioelectroactivity on the denitrification behavior of air-cathode microbial fuel cells

Generally, high bioelectroactivity of anodophilic biofilm favors high power generation of microbial fuel cell (MFC); however, it is not clear whether it can promote denitrification of MFC synchronously. In this study, we studied the impact of anodophilic biofilm bioelectroactivity on the denitrification behavior of air-cathode MFC (AC-MFC) in steady state and found that high bioelectroactivity of anodophilic biofilm not only favored high power generation of the AC-MFC, but also promoted the growth of denitrifers at the anodes and strengthened denitrification. Anodophilic biofilms of AC-MFC with various bioelectroactivity were acclimated at conditions of open circuit (OC), R_ext of 1000 Ω and 20 Ω (denoted as AC-MFC-OC, AC-MFC-1000Ω, and AC-MFC-20Ω, respectively) and performed for over 100 days. Electrochemical tests and microbial analysis results showed that the anode of the AC-MFC-20Ω delivered higher current response of both oxidation and denitrification and had higher abundance of electroactive bacteria than the AC-MFC-OC, AC-MFC-1000Ω, demonstrating a higher bioelectroactivity of the anodophilic biofilms. Moreover, these electroactive bacteria favored the accumulation of denitrifers, like Thauera and Alicycliphilus, probably by consuming trace oxygen through catalyzing oxygen reduction. The AC-MFC-20Ω not only delivered a 61.7% higher power than the AC-MFC-1000Ω, but also achieved a stable and high denitrification rate constant (k_DN) of 1.9 h^?1, which was 50% and 40% higher than that of the AC-MFC-OC and AC-MFC-1000Ω, respectively. It could be concluded that the high bioelectroactivity of the anodophilic biofilms not only favored high power generation of the AC-MFC, but also promoted the enrichment of denitrifers at the anodes and strengthened denitrification. This study provided an effective method for enhancing power generation and denitrification performance of the AC-MFC synchronously.     

Enhancement of electricity production in a mediatorless air–cathode microbial fuel cell using <Emphasis Type="Italic">Klebsiella</Emphasis> sp. IR21

A novel dissimilatory iron-reducing bacteria, Klebsiella sp. IR21, was isolated from the anode biofilm of an MFC reactor. Klebsiella sp. IR21 reduced 27.8 % of ferric iron to ferrous iron demonstrating that Klebsiella sp. IR21 has electron transfer ability. Additionally, Klebsiella sp. IR21 generated electricity forming a biofilm on the anode surface. When a pure culture of Klebsiella sp. IR21 was supplied into a single chamber, air–cathode MFC fed with a mixture of glucose and acetate (500 mg L^?1 COD), 40–60 mV of voltage (17–26 mA m^?2 of current density) was produced. Klebsiella sp. IR21 was also utilized as a biocatalyst to improve the electrical performance of a conventional MFC reactor. A single chamber, air–cathode MFC was fed with reject wastewater (10,000 mg L^?1 COD) from a H₂ fermentation reactor. The average voltage, current density, and power density were 142.9 ± 25.74 mV, 60.5 ± 11.61 mA m^?2, and 8.9 ± 3.65 mW m^?2, respectively, in the MFC without inoculation of Klebsiella sp. IR21. However, these electrical performances of the MFC were significantly increased to 204.7 ± 40.24 mV, 87.5 ± 17.20 mA m^?2, and 18.6 ± 7.23 mW m^?2, respectively, with inoculation of Klebsiella sp. IR21. The results indicate that Klebsiella sp. IR21 can be utilized as a biocatalyst for enhancement of electrical performance in MFC systems.     

Electrocatalytic activity of anodic biofilm responses to pH changes in microbial fuel cells

This study investigates the effects of anodic pH on electricity generation in microbial fuel cells (MFCs) and the intrinsic reasons behind them. In a two-chamber MFC, the maximum power density is 1170 ± 58 mW m⁻² at pH 9.0, which is 29% and 89% higher than those working at pH 7.0 and 5.0, respectively. Electrochemical measurements reveal that pH affects the electron transfer kinetics of anodic biofilms. The apparent electron transfer rate constant (k_app) and exchange current density (i₀) are greater whereas the charge transfer resistance (R_ct) is smaller at pH 9.0 than at other conditions. Scanning electron microscopy verifies that alkaline conditions benefit biofilm formation in MFCs. These results demonstrate that electrochemical interactions between bacteria and electrodes in MFCs are greatly enhanced under alkaline conditions, which can be one of the important reasons for the improved MFC output.     

Glycerol degradation in single-chamber microbial fuel cells

Glycerol degradation with electricity production by a pure culture of Bacillus subtilis in a single-chamber air cathode of microbial fuel cell (MFC) has been demonstrated. Steady state polarization curves indicated a maximum power density of 0.06 mW/cm² with an optimal external resistance of 390Ω. Analysis of the effect of pH on MFC performance demonstrated that electricity generation was sustained over a long period of time under neutral to alkaline conditions. Cyclic voltammetry exhibited the increasing electrochemical activity with the increase of pH of 7, 8 and 9. Voltammetric studies also demonstrated that a two-electron transfer mechanism was occurring in the reactor. The low Coulombic efficiency of 23.08% could be attributed to the loss of electrons for various activities other than electricity generation. This study describes an application of glycerol that could contribute to transformation of the biodiesel industry to a more environmentally friendly microbial fuel cell-based technology.     

A microfluidic microbial fuel cell fabricated by soft lithography

Here we report a new microfluidic microbial fuel cell (MFC) platform built by soft-lithography techniques. The MFC design includes a unique sub-5 μL polydimethylsiloxane soft chamber featuring carbon cloth electrodes and microfluidic delivery of electrolytes. Bioelectricity was generated using Shewanella oneidensis MR-1 cultivated on either complex organic substrates or lactate-based minimal medium. These micro-MFCs exhibited fast start-ups, reproducible current generation, and enhanced power densities up to 62.5 W m⁻³ that represents the best result for sub-100 μL MFCs. Systematic comparisons of custom-made MFC reactors having different chamber sizes indicate volumetric power density is inversely correlated with chamber size in our systems: i.e., the smaller the chamber, the higher the power density is achieved.     

Evaluating the Membrane Fecal Coliform Test by Using Escherichia coli as the Indicator Organism

The fecal coliform membrane filter method (MFC) currently used in water pollution analysis was evaluated by using two strains of Escherichia coli, a known fecal coliform, as the indicator organism. A large relative error in the results obtained with this method was found to be dependent upon the brand of membrane filter employed, the medium, and the temperature of incubation. MFC densities varied between 10 and 60% of the densities determined by means of total bacteria counts and total coliform counts performed at 35 C. Due to the large relative error encountered, the MFC method cannot be recommended as an analytical tool for the laboratory enumeration of E. coli. The results do show that the MFC method can be used at 35 C for enumeration of E. coli and for differential counts of E. coli and Enterobacter aerogenes.     

Effect of nitrogen addition on the performance of microbial fuel cell anodes

Carbon cloth anodes were modified with 4(N,N-dimethylamino)benzene diazonium tetrafluoroborate to increase nitrogen-containing functional groups at the anode surface in order to test whether the performance of microbial fuel cells (MFCs) could be improved by controllably modifying the anode surface chemistry. Anodes with the lowest extent of functionalization, based on a nitrogen/carbon ratio of 0.7 as measured by XPS, achieved the highest power density of 938 mW/m². This power density was 24% greater than an untreated anode, and similar to that obtained with an ammonia gas treatment previously shown to increase power. Increasing the nitrogen/carbon ratio to 3.8, however, decreased the power density to 707 mW/m². These results demonstrate that a small amount of nitrogen functionalization on the carbon cloth material is sufficient to enhance MFC performance, likely as a result of promoting bacterial adhesion to the surface without adversely affecting microbial viability or electron transfer to the surface.     

不同林龄尾巨桉人工林的生物量及其分配特征

根据1,2,3,5,8a共5个不同年龄的15块1000 m2尾巨桉样地(3次重复)调查资料,利用18株不同年龄和径阶的样木数据,建立以胸径(D)为单变量的生物量回归方程。采用样木回归分析法(乔木层)和样方收获法(灌木层、草本层、地上凋落物)获取不同林龄尾巨桉人工林的生物量,分析了其组成、分配及不同林龄生物量的变化趋势。结果表明:林分总生物量随林龄而增加,1,2,3,5年生和8年生尾巨桉人工林生物量分别为12.49,47.75,64.51,105.77和137.51 t/hm2,其中活体植物占85.60%—97.61%,地上凋落物占2.39%—14.40%;层次分配方面乔木层占绝对优势,占54.80%—91.56%,且随林龄的增加而增大,其次为凋落物,灌木层和草本层生物量较小,分别占1.02%—6.47%和0.28%—24.33%,均随林龄的增加呈递减趋势;乔木层以干所占比例最高,占51.07%—98.48%,且随林龄而增加,枝、叶、根分别占5.76%—11.80%,2.17%—21.01%和6.72%—14.87%,均随林龄而下降;灌木层以枝所占比例最高,为37.89%—56.79%,叶和根分别占16.35%—34.24%和19.52%—39.52%,随林龄的变化均不大;草本层分配1—5年生以地上所占比例较大,8年生地下所占比例高达63.87%;尾巨桉人工林乔木层各器官、地上凋落物及总生物量具有良好的优化增长模型,其总生物量的增长模型为Y=-1.693×104+3.337×104X-1.761X2;8年生尾巨桉人工林总生物量与30年生的木莲人工林持平,低于热带雨林,但其年均净生产量高达17.19 t/hm2,是一个光合效率高、固碳潜力大的速生丰产优良造林树种。     

Acetate oxidation by syntrophic association between Geobacter sulfurreducens and a hydrogen-utilizing exoelectrogen

Anodic microbial communities in acetate-fed microbial fuel cells (MFCs) were analyzed using stable-isotope probing of 16S rRNA genes followed by denaturing gradient gel electrophoresis. The results revealed that Geobacter sulfurreducens and Hydrogenophaga sp. predominated in the anodic biofilm. Although the predominance of Geobacter sp. as acetoclastic exoelectrogens in acetate-fed MFC systems has been often reported, the ecophysiological role of Hydrogenophaga sp. is unknown. Therefore, we isolated and characterized a bacterium closely related to Hydrogenophaga sp. (designated strain AR20). The newly isolated strain AR20 could use molecular hydrogen (H₂), but not acetate, with carbon electrode as the electron acceptor, indicating that the strain AR20 was a hydrogenotrophic exoelectrogen. This evidence raises a hypothesis that acetate was oxidized by G. sulfurreducens in syntrophic cooperation with the strain AR20 as a hydrogen-consuming partner in the acetate-fed MFC. To prove this hypothesis, G. sulfurreducens strain PCA was cocultivated with the strain AR20 in the acetate-fed MFC without any dissolved electron acceptors. In the coculture MFC of G. sulfurreducens and strain AR20, current generation and acetate degradation were the highest, and the growth of strain AR20 was observed. No current generation, acetate degradation and cell growth occurred in the strain AR20 pure culture MFC. These results show for the first time that G. sulfurreducens can oxidize acetate in syntrophic cooperation with the isolated Hydrogenophaga sp. strain AR20, with electrode as the electron acceptor.     

High shear enrichment improves the performance of the anodophilic microbial consortium in a microbial fuel cell

In many microbial bioreactors, high shear rates result in strong attachment of microbes and dense biofilms. In this study, high shear rates were applied to enrich an anodophilic microbial consortium in a microbial fuel cell (MFC). Enrichment at a shear rate of about 120 s^?1 resulted in the production of a current and power output two to three times higher than those in the case of low shear rates (around 0.3 s^?1). Biomass and biofilm analyses showed that the anodic biofilm from the MFC enriched under high shear rate conditions, in comparison with that under low shear rate conditions, had a doubled average thickness and the biomass density increased with a factor 5. The microbial community of the former, as analysed by DGGE, was significantly different from that of the latter. The results showed that enrichment by applying high shear rates in an MFC can result in a specific electrochemically active biofilm that is thicker and denser and attaches better, and hence has a better performance.     

Dianthus caryophyllus stems and Zantedeschia aethiopica petioles/pedicels show anatomical features indicating efficient photosynthesis

The fine structure of the green stem of Dianthus caryophyllus, the leaf petiole and the flower pedicel of Zantedeschia aethiopica were studied using light and scanning electron microscopy. It was revealed that these non-foliar plant parts of both species possess epidermis with numerous stomata. Stomatal density of D. caryophyllus stem was found to be relatively high (79 vs 100 per mm² found on leaf surface). Z. aethiopica petioles and pedicels also possess numerous stomata (17 per mm²), yet stomatal density was found to be about half of that of leaves. Anatomical differences observed between petioles and pedicels were only minor. Stems of D. caryophyllus as well as petioles and pedicels of Z. aethiopica have a chlorenchyma-type tissue whose fine structure is quite similar to the leaf palisade chlorenchyma. Yet, the palisade of Z. aethiopica petioles and pedicels shows a peculiar arrangement: palisade cells are arranged with their long axis parallel to the longitudinal organ axis. Palisade tissue found in the aforementioned non-foliar plant organs in both species shows strong red chlorophyll auto-fluorescence under epi-fluorescence optics, consists of cells with abundant chloroplasts, possesses high percentage of intercellular spaces (13 and 20%, respectively) and its cells expose considerable part of their surface to the intercellular air. The fine structure of this stem palisade tissue along with the abundance of functional stomata found on the epidermis may support efficient photosynthesis.     

Continuous power generation and microbial community structure of the anode biofilms in a three-stage microbial fuel cell system

A mediator-less three-stage two-chamber microbial fuel cell (MFC) system was developed and operated continuously for more than 1.5 years to evaluate continuous power generation while treating artificial wastewater containing glucose (10 mM) concurrently. A stable power density of 28 W/m³ was attained with an anode hydraulic retention time of 4.5 h and phosphate buffer as the cathode electrolyte. An overall dissolved organic carbon removal ratio was about 85%, and coulombic efficiency was about 46% in this MFC system. We also analyzed the microbial community structure of anode biofilms in each MFC. Since the environment in each MFC was different due to passing on the products to the next MFC in series, the microbial community structure was different accordingly. The anode biofilm in the first MFC consisted mainly of bacteria belonging to the Gammaproteobacteria, identified as Aeromonas sp., while the Firmicutes dominated the anode biofilms in the second and third MFCs that were mainly fed with acetate. Cyclic voltammetric results supported the presence of a redox compound(s) associated with the anode biofilm matrix, rather than mobile (dissolved) forms, which could be responsible for the electron transfer to the anode. Scanning electron microscopy revealed that the anode biofilms were comprised of morphologically different cells that were firmly attached on the anode surface and interconnected each other with anchor-like filamentous appendages, which might support the results of cyclic voltammetry. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Investigating microbial fuel cell bioanode performance under different cathode conditions

A compact, three‐in‐one, flow‐through, porous, electrode design with minimal electrode spacing and minimal dead volume was implemented to develop a microbial fuel cell (MFC) with improved anode performance. A biofilm‐dominated anode consortium enriched under a multimode, continuous‐flow regime was used. The increase in the power density of the MFC was investigated by changing the cathode (type, as well as catholyte strength) to determine whether anode was limiting. The power density obtained with an air‐breathing cathode was 56 W/m³ of net anode volume (590 mW/m²) and 203 W/m³ (2160 mW/m²) with a 50‐mM ferricyanide‐based cathode. Increasing the ferricyanide concentration and ionic strength further increased the power density, reaching 304 W/m³ (3220 mW/m², with 200 mM ferricyanide and 200 mM buffer concentration). The increasing trend in the power density indicated that the anode was not limiting and that higher power densities could be obtained using cathodes capable of higher rates of oxidation. The internal solution resistance for the MFC was 5–6 Ω, which supported the improved performance of the anode design. A new parameter defined as the ratio of projected surface area to total anode volume is suggested as a design parameter to relate volumetric and area‐based power densities and to enable comparison of various MFC configurations. Published 2009 American Institute of Chemical Engineers Biotechnol. Prog., 2009     

Evaluation of procedures to acclimate a microbial fuel cell for electricity production

A microbial fuel cell (MFC) is a relatively new type of fixed film bioreactor for wastewater treatment, and the most effective methods for inoculation are not well understood. Various techniques to enrich electrochemically active bacteria on an electrode were therefore studied using anaerobic sewage sludge in a two-chambered MFC. With a porous carbon paper anode electrode, 8 mW/m² of power was generated within 50 h with a Coulombic efficiency (CE) of 40%. When an iron oxide-coated electrode was used, the power and the CE reached 30 mW/m² and 80%, respectively. A methanogen inhibitor (2-bromoethanesulfonate) increased the CE to 70%. Bacteria in sludge were enriched by serial transfer using a ferric iron medium, but when this enrichment was used in a MFC the power was lower (2 mW/m²) than that obtained with the original inoculum. By applying biofilm scraped from the anode of a working MFC to a new anode electrode, the maximum power was increased to 40 mW/m². When a second anode was introduced into an operating MFC the acclimation time was not reduced and the total power did not increase. These results suggest that these active inoculating techniques could increase the effectiveness of enrichment, and that start up is most successful when the biofilm is harvested from the anode of an existing MFC and applied to the new anode.     

Role of galE on biofilm formation by Thermus spp.

Thermus thermophilus and Thermus aquaticus are thermophilic bacteria that are frequently found to attach to solid surfaces in hot springs to form biofilms. Uridine diphosphate (UDP)-galactose-4′-epimerase (GalE) is an enzyme that catalyzes the conversion of UDP-galactose to UDP-glucose, an important biochemical step in exopolysaccharide synthesis. We expressed GalE obtained from T. thermophilus HB8 in Escherichia coli and found that the enzyme is stable at 80 °C and can epimerize UDP-galactose to UDP-glucose and UDP-N-acetylgalactosamine (UDP-GalNAc) to UDP-N-acetylglucosamine (UDP-GlcNAc). Enzyme overexpression in T. thermophilus HB27 led to an increased capacity of biofilm production. Therefore, the galE gene is important to biofilm formation because of its involvement in epimerizing UDP-galactose and UDP-N-acetylgalactosamine for exopolysaccharide biosynthesis.     

Recovery of electric energy from formate by using a recombinant strain of Escherichia coli

Recombinant Escherichia coli cells were applied for the recovery of electric energy from formate. Initially, the fdh gene, which encodes formate dehydrogenase (FDH) of Mycobacterium vaccae, was introduced into E. coli cells to allow efficient degradation of formate. The constructed microbial fuel cell (MFC) with E. coli BW25113 cells carrying fdh gene showed appreciable generation of current density in the presence of formate as a substrate. Current density and polarization curves revealed that the performance of MFC under examined conditions was limited by the electron transfer from bulk liquid to the electrode surface; accordingly, agitation resulted in an increase in the current density and achieved a coulombic efficiency of 21.7 % on the basis of formate consumed. Thus, gene recombination enables E. coli cells to utilize formate as a fuel for MFC.