Modelling potential/current distribution in microbial electrochemical systems shows how the optimal bioanode architecture depends on electrolyte conductivity.

The theoretical bases for modelling the distribution of the electrostatic potential in microbial electrochemical systems are described. The secondary potential distribution (i.e. without mass transport limitation of the substrate) is shown to be sufficient to validly address microbial electrolysis cells (MECs). MECs are modelled with two different ionic conductivities of the solution (1 and 5.3 S m(-1)) and two bioanode kinetics (jmax = 5.8 or 34 A m(-2)). A conventional reactor configuration, with the anode and the cathode face to face, is compared with a configuration where the bioanode perpendicular to the cathode implements the electrochemical reaction on its two sides. The low solution conductivity is shown to have a crucial impact, which cancels out the advantages obtained by setting the bioanode perpendicular to the cathode. For the same reason, when the surface area of the anode is increased by multiplying the number of plates, care must be taken not to create too dense anode architecture. Actually, the advantages of increasing the surface area by multiplying the number of plates can be lost through worsening of the electrochemical conditions in the multi-layered anode, because of the increase of the electrostatic potential of the solution inside the anode structure. The model gives the first theoretical bases for scaling up MECs in a rather simple but rigorous way.

Towards an engineering-oriented strategy for building microbial anodes for microbial fuel cells.

The objective of the work was to give some first insight into an engineering-oriented approach to MFC design by focusing on anode optimisation. The effect of various parameters was firstly investigated in half cell set-ups under well-controlled conditions. Microbial anodes were formed from soil leachate under polarisation at -0.2 V vs. SCE with different concentrations of substrate, salt and buffer. It was shown that non-turnover CV could be used to assess the electroactive maturity of the anodes during polarisation. This first phase resulted in the definition of a set of optimal parameter values. In the second phase, an optimal anode was formed in a half-cell under the defined optimal conditions. A numerical approach was then developed to calculate the theoretical maximum power that the anode could provide in an ideal MFC. The concept of "ideal MFC" introduced here allowed the theoretical maximum power to be calculated on the sole basis of the kinetic characteristics of the anode. Finally, a MFC designed in the aim of approaching such ideal conditions generated stable power densities of 6.0 W m(-2), which were among the highest values reported so far. The discrepancy between the theoretical maximum (8.9 W m(-2)) and the experimental results pointed out some limit due to the source of inoculum and suggested possible paths to improvement.

Oxygen-reducing microbial cathodes in hypersaline electrolyte.

Hypersaline electrolytes offer a way to boost the development of microbial fuel cells by overcoming the issue due to the low conductivity of the usual media. Efficient halotolerant bioanodes have already been designed but O2-reducing cathodes remain a strong bottleneck. Here, O2-reducing biocathodes were designed by using salt marsh sediment as the inoculum and a hypersaline media (45 g/L NaCl) of high conductivity (10.4 S m-1). Current density up to 2.2 A m-2 was reached from potential of +0.2 V/SCE. The efficiency of the biocathodes was correlated to the presence of Gammaproteobacteria strain(s) related to Thiohalobacter thiocyanaticus, which were considerably enriched in the best performing biocathodes. This work opens up new perspectives to overcome the O2 reduction issue in hypersaline MFCs by designing efficient halotolerant microbial cathodes and pointing out the strains that should now be focused to improve them.

Hypersaline microbial fuel cell equipped with an oxygen-reducing microbial cathode.

Microbial anodes and oxygen reducing microbial cathodes were designed separately under constant polarization at + 0.1 V/SCE in a hypersaline medium (NaCl 45 g/L). They were then associated to design two-compartment microbial fuel cells (MFCs). These MFCs produced up to 209 ± 24 mW m-2 during a week. This was the first demonstration that hypersaline MFCs equipped with microbial cathodes can produce power density at this level. Desulfuromonas sp. were confirmed to be key species of the anodes. The efficiency of the cathodes was linked to the development of a redox system centred at + 0.2 V/SCE and to the presence of Gammaproteobacteria (Alteromonadales and Oceanospirillales), especially an unclassified order phylogenetically linked to the genus Thioalobacter. Comparing the different performance of the four MFCs with the population analyses suggested that polarization at + 0.1 V/SCE should be maintained longer to promote the growth of Thioalobacter on the cathode and thus increase the MFC performance.

Effect of surface roughness, porosity and roughened micro-pillar structures on the early formation of microbial anodes.

The early formation of electroactive biofilms was investigated with gold electrodes inoculated with Geobacter sulfurreducens. Biofilms were formed under an applied potential of 0.1 V/SCE, with a single batch of acetate 10 mM, on flat gold electrodes with different random surface roughness. Roughness with arithmetical mean height (Sa) ranging from 0.5 to 6.7 µm decreased the initial latency time, and increased the current density by a factor of 2.7 to 6.7 with respect to nano-rough electrodes (Sa = 4.5 nm). The current density increased linearly with Sa up to 14.0 A·m-2 for Sa of 6.7 µm. This linear relationship remained valid for porous gold. In this case, the biofilm rapidly formed a uniform layer over the pores, so porosity impacted the current only by modifying the roughness of the upper surface. The current density thus reached 14.8 ±â€¯1.1 A·m-2 with Sa of 7.6 µm (7 times higher than the nano-rough electrodes). Arrays of 500-µm-high micro-pillars were roughened following the same protocol. In this case, roughening resulted in a modest gain around 1.3-fold. A numerical model showed that the modest enhancement was due to ion transport not being sufficient to mitigate the local acidification of the structure bottom.

Fungal contamination and Aflatoxin B1 and Ochratoxin A in Lebanese wine-grapes and musts.

Five hundred and ten strains of filamentous fungi were isolated from Lebanese grapes during 2005 at veraison and harvesting periods. Four hundred eighty-seven isolates belonged to the Aspergillus spp. (95.5%) and 23 belonged to the Penicillium spp. (4.5%). Black aspergilli constituted 56.9% (52.2% Aspergillus niger aggregates, 2.9% Aspergillus japonicus and 1.8% Aspergillus carbonarius) while the isolation rate of Aspergillus flavus the none habitual member of grape mycobiota was 43.1% of the total Aspergillus spp. isolated. All isolates were tested for the ability to produce the Ochratoxin A (OTA) and the Aflatoxin B1 (AFB1). A. carbonarius showed that it is the only species able to produce the OTA with a production ability of 100% and a maximum concentration reaching 8.38microg/g CYA. As for the aflatoxigenic ability, 43.4% of A. flavus isolates produced this mycotoxin with a maximum production reaching 22.6microg/g CYA while none of the other isolates showed a production capacity of this mycotoxin. Forty-seven samples of must produced from the collected grapes were also analyzed. None of these samples was contaminated by OTA at a detectable limit while 40% of these same samples were found to contain AFB1 with concentrations ranging from 0.01 to 0.46microgl(-1).

Forming electrochemically active biofilms from garden compost under chronoamperometry.

Dimensionally stable anodes (DSA) were polarized at different constant potential values for several days in garden compost. After an initial lag period ranging from 1 to 10.5 days, the current increased fast and then stabilized for days. Current densities higher than 100 mA m(-2) and up to 385 mA m(-2) were obtained with the sole organic matter contained in compost as substrate. Control experiments performed with sterilized compost, oscillations of the current with the temperature, kinetics of the exponential phase of current increase and observations of the surface of electrodes by epifluorescence microscopy showed that the current was controlled by the colonization of the electrode surface by a biofilm which originated the indigenous flora of compost. Three individually addressed electrodes polarized at different potentials in the same reactor led to identical current evolutions on each electrode, which underlined the key role of the microbial flora of the compost in the discrepancy observed in the other experiments. Chronoamperometry revealed a promising technique to check natural environments for new electrochemically active microbial species.

Impact of electrode micro- and nano-scale topography on the formation and performance of microbial electrodes.

From a fundamental standpoint, microbial electrochemistry is unravelling a thrilling link between life and materials. Technically, it may be the source of a large number of new processes such as microbial fuel cells for powering remote sensors, autonomous sensors, microbial electrolysers and equipment for effluent treatment. Microbial electron transfers are also involved in many natural processes such as biocorrosion. In these contexts, a huge number of studies have dealt with the impact of electrode materials, coatings and surface functionalizations but very few have focused on the effect of the surface topography, although it has often been pointed out as a key parameter impacting the performance of electroactive biofilms. The first part of the review gives an overview of the influence of electrode topography on abiotic electrochemical reactions. The second part recalls some basics of the effect of surface topography on bacterial adhesion and biofilm formation, in a broad domain reaching beyond the context of electroactivity. On these well-established bases, the effect of surface topography is reviewed and analysed in the field of electroactive biofilms. General trends are extracted and fundamental questions are pointed out, which should be addressed to boost future research endeavours. The objective is to provide basic guidelines useful to the widest possible range of research communities so that they can exploit surface topography as a powerful lever to improve, or to mitigate in the case of biocorrosion for instance, the performance of electrode/biofilm interfaces.

Effect of surface nano/micro-structuring on the early formation of microbial anodes with Geobacter sulfurreducens: Experimental and theoretical approaches.

Smooth and nano-rough flat gold electrodes were manufactured with controlled Ra of 0.8 and 4.5nm, respectively. Further nano-rough surfaces (Ra 4.5nm) were patterned with arrays of micro-pillars 500µm high. All these electrodes were implemented in pure cultures of Geobacter sulfurreducens, under a constant potential of 0.1V/SCE and with a single addition of acetate 10mM to check the early formation of microbial anodes. The flat smooth electrodes produced an average current density of 0.9A·m-2. The flat nano-rough electrodes reached 2.5A·m-2 on average, but with a large experimental deviation of ±2.0A·m-2. This large deviation was due to the erratic colonization of the surface but, when settled on the surface, the cells displayed current density that was directly correlated to the biofilm coverage ratio. The micro-pillars considerably improved the experimental reproducibility by offering the cells a quieter environment, facilitating biofilm development. Current densities of up to 8.5A·m-2 (per projected surface area) were thus reached, in spite of rate limitation due to the mass transport of the buffering species, as demonstrated by numerical modelling. Nano-roughness combined with micro-structuring increased current density by a factor close to 10 with respect to the smooth flat surface.

Occurrence of ochratoxin A- and aflatoxin B1-producing fungi in Lebanese grapes and ochratoxin a content in musts and finished wines during 2004.

This paper reports the results of an extensive survey on the occurrence of filamentous fungi isolated from wine-grapes in Lebanon and to test their ability to produce ochratoxin A (OTA) and aflatoxin B1 (AFB1) on CYA culture medium, in order to assess their potential for producing these mycotoxins on grapes. From the 470 grapes samples taken during season 2004, 550 fungi strains were isolated with 490 belonging to Aspergillus spp. and 60 belonging to Penicillium spp. All these isolated fungi starins were tested for their ability to produce OTA and AFB1. Aspergillus carbonarius shows that it is the only species able to produce OTA with a production percentage reaching 100% and a maximum concentration of 52.8 microg/g of Czapek yeast extract agar (CYA). In its turn, Aspergillus flavus was considered as the only AFB1-producing species with production percentage of 45.3% and a maximum concentration reaching 40 microg/g CYA. A total of 47 handmade musts produced from the collected grapes were also analyzed in order to correlate the presence of OTA in must and the occurrence of filamentous fungi on grapes; 57.4% were contaminated with OTA at low level with concentrations ranging between 0.011 and 0.221 microg OTA L(-1). The analysis of these must samples was not performed with regard to AFB1. Seventy samples of finish red wine were also assayed for OTA content. The results showed that 42 of the tested samples (60%) were found to be positive for OTA with low levels (0.012-0.126 microg OTA L(-1)).

Halotolerant bioanodes: The applied potential modulates the electrochemical characteristics, the biofilm structure and the ratio of the two dominant genera.

The development of economically-efficient microbial electrochemical technologies remains hindered by the low ionic conductivity of the culture media used as the electrolyte. To overcome this drawback, halotolerant bioanodes were designed with salt marsh sediment used as the inoculum in electrolytes containing NaCl at 30 or 45g/L (ionic conductivity 7.0 or 10.4S·m(-1)). The bioanodes were formed at four different potentials -0.4, -0.2, 0.0 and 0.2V/SCE to identify the effect on the electrochemical kinetic parameters, the biofilm structures and the composition of the microbial communities. The bioanodes formed at -0.4V/SCE were largely dominated by Marinobacter spp. Voltammetry showed that they provided higher currents than the other bioanodes in the range of low potentials, but the maximum currents were limited by the poor surface colonization. The bioanodes formed at -0.2, 0.0 and 0.2V/SCE showed similar ratios of Marinobacter and Desulfuromonas spp. and higher values of the maximum current density. The combined analysis of kinetic parameters, biofilm structure and biofilm composition showed that Marinobacter spp., which ensured a higher electron transfer rate, were promising species for the design of halotolerant bioanodes. The challenge is now to overcome its limited surface colonization in the absence of Desulfuromonas spp.

Electrochemical characterization of microbial bioanodes formed on a collector/electrode system in a highly saline electrolyte.

Bioanodes were formed with electrodes made of carbon felt and equipped with a titanium electrical collector, as commonly used in microbial fuel cells. Electrochemical impedance spectroscopy (EIS) performed on the abiotic electrode system evidenced two time constants, one corresponding to the "collector/carbon felt" contact, the other to the "carbon felt/solution" interface. Such a two time constant system was characteristics of the two-material electrode, independent of biofilm presence. EIS was then performed during the bioanode formation around the constant applied potential of 0.1 V/SCE. The equivalent electrical model was similar to that of the abiotic system. Due to the high salinity of the electrolyte (45 g·L(-1) NaCl) the electrolyte resistance was always very low. The bioanode development induced kinetic heterogeneities that were taken into account by replacing the pure capacitance of the abiotic system by a constant phase element for the "carbon felt/solution" interface. The current increase from 0 to 20.6 A·m(-2) was correlated to the considerable decrease of the charge transfer resistance of the "carbon felt/solution" interface from 2.4 10(4) to 92 Ω·cm(2). Finally, EIS implemented at 0.4 V/SCE showed that the limitation observed at high potential values was not related to mass transfer but to a biofilm-linked kinetics.

Forming microbial anodes under delayed polarisation modifies the electron transfer network and decreases the polarisation time required.

Microbial anodes were formed from compost leachate on carbon cloth electrodes. The biofilms formed at the surface of electrodes kept at open circuit contained microorganisms that switched their metabolism towards electrode respiration in response to a few minutes of polarisation. When polarisation at -0.2 V/SCE (+0.04 V/SHE) was applied to a pre-established biofilm formed at open circuit (delayed polarisation), the bacteria developed an extracellular electron transport network that showed multiple redox systems, reaching 9.4 A/m(2) after only 3-9 days of polarisation. In contrast, when polarisation was applied from the beginning, bacteria developed a well-tuned extracellular electron transfer network concomitantly with their growth, but 36 days of polarisation were required to get current of the same order (6-8 A/m(2)). The difference in performance was attributed to the thinner, more heterogeneous structure of the biofilms obtained by delayed polarisation compared to the thick uniform structure obtained by full polarisation.

Harvesting electricity with Geobacter bremensis isolated from compost.

Electrochemically active (EA) biofilms were formed on metallic dimensionally stable anode-type electrode (DSA), embedded in garden compost and polarized at +0.50 V/SCE. Analysis of 16S rRNA gene libraries revealed that biofilms were heavily enriched in Deltaproteobacteria in comparison to control biofilms formed on non-polarized electrodes, which were preferentially composed of Gammaproteobacteria and Firmicutes. Among Deltaproteobacteria, sequences affiliated with Pelobacter and Geobacter genera were identified. A bacterial consortium was cultivated, in which 25 isolates were identified as Geobacter bremensis. Pure cultures of 4 different G. bremensis isolates gave higher current densities (1400 mA/m(2) on DSA, 2490 mA/m(2) on graphite) than the original multi-species biofilms (in average 300 mA/m(2) on DSA) and the G. bremensis DSM type strain (100-300 A/m(2) on DSA; 2485 mA/m(2) on graphite). FISH analysis confirmed that G. bremensis represented a minor fraction in the original EA biofilm, in which species related to Pelobacter genus were predominant. The Pelobacter type strain did not show EA capacity, which can explain the lower performance of the multi-species biofilms. These results stressed the great interest of extracting and culturing pure EA strains from wild EA biofilms to improve the current density provided by microbial anodes.

Catalysis of the electrochemical reduction of oxygen by bacteria isolated from electro-active biofilms formed in seawater.

Biofilms formed in aerobic seawater on stainless steel are known to be efficient catalysts of the electrochemical reduction of oxygen. Based on their genomic analysis, seven bacterial isolates were selected and a cyclic voltammetry (CV) procedure was implemented to check their electrocatalytic activity towards oxygen reduction. All isolates exhibited close catalytic characteristics. Comparison between CVs recorded with glassy carbon and pyrolytic graphite electrodes showed that the catalytic effect was not correlated with the surface area covered by the cells. The low catalytic effect obtained with filtered isolates indicated the involvement of released redox compounds, which was confirmed by CVs performed with adsorbed iron-porphyrin. None of the isolates were able to form electro-active biofilms under constant polarization. The capacity to catalyze oxygen reduction is shown to be a widespread property among bacteria, but the property detected by CV does not necessarily confer the ability to achieve stable oxygen reduction under constant polarization.

Testing various food-industry wastes for electricity production in microbial fuel cell.

Three food-industry wastes: fermented apple juice (FAJ), wine lees and yogurt waste (YW) were evaluated in combination with two sources of inoculum, anaerobic sludge and garden compost, to produce electricity in microbial fuel cells. Preliminary potentiostatic studies suggested that YW was the best candidate, able to provide up to 250 mA/m(2) at poised potential +0.3V/SCE. Experiments conducted with two-chamber MFCs confirmed that wine lees were definitely not suitable. FAJ was not able to start an MFC by means of its endogenous microflora, while YW was. Both FAJ and YW were suitable fuels when anaerobic sludge or compost leachate was used as inoculum source. Sludge-MFCs had better performance using YW (54 mW/m(2) at 232 mA/m(2)). In contrast, compost-leachate MFCs showed higher power density with FAJ (78 mW/m(2) at 209 mA/m(2)) than with YW (37 mW/m(2) at 144 mA/m(2)) but YW gave more stable production. Under optimized operating conditions, compost-leachate MFCs fueled with YW gave up to 92 mW/m(2) at 404 mA/m(2) and 44 mW/m(2) in stable conditions.

Marine aerobic biofilm as biocathode catalyst.

Stainless steel electrodes were immersed in open seawater and polarized for some days at -200 mV vs. Ag/AgCl. The current increase indicated the formation of biofilms that catalysed the electrochemical reduction of oxygen. These wild, electrochemically active (EA) biofilms were scraped, resuspended in seawater and used as the inoculum in closed 0.5L electrochemical reactors. This procedure allowed marine biofilms that are able to catalyse oxygen reduction to be formed in small, closed small vessels for the first time. Potential polarisation during biofilm formation was required to obtain EA biofilms and the roughness of the surface favoured high current values. The low availability of nutrients was shown to be a main limitation. Using an open reactor continuously fed with filtered seawater multiplied the current density by a factor of around 20, up to 60 microA/cm(2), which was higher than the current density provided in open seawater by the initial wild biofilm. These high values were attributed to continuous feeding with the nutrients contained in seawater and to suppression of the indigenous microbial species that compete with EA strains in natural open environments. Pure isolates were extracted from the wild biofilms and checked for EA properties. Of more than thirty different species tested, only Winogradskyella poriferorum and Acinetobacter johsonii gave current densities of respectively 7% and 3% of the current obtained with the wild biofilm used as inoculum. Current densities obtained with pure cultures were lower than those obtained with wild biofilms. It is suspected that synergic effects occur in whole biofilms or/and that wild strains may be more efficient than the cultured isolates.

Decolorization of synthetic melanoidins-containing wastewater by a bacterial consortium.

The presence of melanoidins in molasses wastewater leads to water pollution both due to its dark brown color and its COD contents. In this study, a bacterial consortium isolated from waterfall sediment was tested for its decolorization. The identification of culturable bacteria by 16S rDNA based approach showed that the consortium composed of Klebsiella oxytoca, Serratia mercescens, Citrobacter sp. and unknown bacterium. In the context of academic study, prevention on the difficulties of providing effluent as well as its variations in compositions, several synthetic media prepared with respect to color and COD contents based on analysis of molasses wastewater, i.e., Viandox sauce (13.5% v/v), caramel (30% w/v), beet molasses wastewater (41.5% v/v) and sugarcane molasses wastewater (20% v/v) were used for decolorization using consortium with color removal 9.5, 1.13, 8.02 and 17.5%, respectively, within 2 days. However, Viandox sauce was retained for further study. The effect of initial pH and Viandox concentration on decolorization and growth of bacterial consortium were further determined. The highest decolorization of 18.3% was achieved at pH 4 after 2 day of incubation. Experiments on fresh or used medium and used or fresh bacterial cells, led to conclusion that the limitation of decolorization was due to nutritional deficiency. The effect of aeration on decolorization was also carried out in 2 L laboratory-scale suspended cell bioreactor. The maximum decolorization was 19.3% with aeration at KLa=2.5836 h(-1) (0.1 vvm).

Effect of temperature on Brettanomyces bruxellensis: metabolic and kinetic aspects.

The effect of temperatures ranging from 15 to 35 degrees C on a culture of Brettanomyces bruxellensis was investigated in regards to thermodynamics, metabolism, and kinetics. In this temperature range, we observed an increase in growth and production rates. The growth behavior was well represented using the Arrhenius model, and an apparent activation energy of 16.61 kcal/mol was estimated. A stuck fermentation was observed at 35 degrees C as represented by high cell death. The carbon balance established that temperature had no effect on repartition of the glucose consumption between biomass and products. Hence, the same biomass concentration was obtained for all temperatures, except at 35 degrees C. Moreover, using logistic and Luedeking-Piret models, we demonstrated that production rates of ethanol and acetic acid were partially growth associated. Parameters associated with growth (alpha eth and alpha aa) remained constant with changing temperature, whereas, parameters associated with the population (beta eth and beta aa) varied. Optimal values were obtained at 32 degrees C for ethanol and at 25 degrees C for acetic acid.