Nitrogen Fixation and Ammonium Assimilation Pathway Expression of Geobacter sulfurreducens Changes in Response to the Anode Potential in Microbial Electrochemical Cells.

Nitrogen gas (N2) fixation in the anode-respiring bacterium Geobacter sulfurreducens occurs through complex, multistep processes. Optimizing ammonium (NH4+) production from this bacterium in microbial electrochemical technologies (METs) requires an understanding of how those processes are regulated in response to electrical driving forces. In this study, we quantified gene expression levels (via RNA sequencing) of G. sulfurreducens growing on anodes fixed at two different potentials (-0.15 V and +0.15 V versus standard hydrogen electrode). The anode potential had a significant impact on the expression levels of N2 fixation genes. At -0.15 V, the expression of nitrogenase genes, such as nifH, nifD, and nifK, significantly increased relative to that at +0.15 V, as well as genes associated with NH4+ uptake and transformation, such as glutamine and glutamate synthetases. Metabolite analysis confirmed that both of these organic compounds were present in significantly higher intracellular concentrations at -0.15 V. N2 fixation rates (estimated using the acetylene reduction assay and normalized to total protein) were significantly larger at -0.15 V. Genes expressing flavin-based electron bifurcation complexes, such as electron-transferring flavoproteins (EtfAB) and the NADH-dependent ferredoxin:NADP reductase (NfnAB), were also significantly upregulated at -0.15 V, suggesting that these mechanisms may be involved in N2 fixation at that potential. Our results show that in energy-constrained situations (i.e., low anode potential), the cells increase per-cell respiration and N2 fixation rates. We hypothesize that at -0.15 V, they increase N2 fixation activity to help maintain redox homeostasis, and they leverage electron bifurcation as a strategy to optimize energy generation and use. IMPORTANCE Biological nitrogen fixation coupled with ammonium recovery provides a sustainable alternative to the carbon-, water-, and energy-intensive Haber-Bosch process. Aerobic biological nitrogen fixation technologies are hindered by oxygen gas inhibition of the nitrogenase enzyme. Electrically driving biological nitrogen fixation in anaerobic microbial electrochemical technologies overcomes this challenge. Using Geobacter sulfurreducens as a model exoelectrogenic diazotroph, we show that the anode potential in microbial electrochemical technologies has a significant impact on nitrogen gas fixation rates, ammonium assimilation pathways, and expression of genes associated with nitrogen gas fixation. These findings have important implications for understanding regulatory pathways of nitrogen gas fixation and will help identify target genes and operational strategies to enhance ammonium production in microbial electrochemical technologies.

Perspective: Phosphorus monitoring must be rooted in sustainability frameworks spanning material scale to human scale.

Phosphorus (P) is a finite resource, and its environmental fate and transport is complex. With fertilizer prices expected to remain high for years and disruption to supply chains, there is a pressing need to recover and reuse P (primarily as fertilizer). Whether recovery is to occur from urban systems (e.g., human urine), agricultural soil (e.g., legacy P), or from contaminated surface waters, quantification of P in various forms is vital. Monitoring systems with embedded near real time decision support, so called cyber physical systems, are likely to play a major role in the management of P throughout agro-ecosystems. Data on P flow(s) connects the environmental, economic, and social pillars of the triple bottom line (TBL) sustainabilty framework. Emerging monitoring systems must account for complex interactions in the sample, and interface with a dynamic decision support system that considers adaptive dynamics to societal needs. It is known from decades of study that P is ubiquitous, yet without quantitative tools for studying the dynamic nature of P in the environment, the details may remain elusive. If new monitoring systems (including CPS and mobile sensors) are informed by sustainability frameworks, data-informed decision making may foster resource recovery and environmental stewardship from technology users to policymakers.

Mechanisms of orthophosphate removal from water by lanthanum carbonate and other lanthanum-containing materials.

Removing phosphorus (P) from water and wastewater is essential for preventing eutrophication and protecting environmental quality. Lanthanum [La(III)]-containing materials can effectively and selectively remove orthophosphate (PO4) from aqueous systems, but there remains a need to better understand the underlying mechanism of PO4 removal. Our objectives were to 1) identify the mechanism of PO4 removal by La-containing materials and 2) evaluate the ability of a new material, La2(CO3)3(s), to remove PO4 from different aqueous matrices, including municipal wastewater. We determined the dominant mechanism of PO4 removal by comparing geochemical simulations with equilibrium data from batch experiments and analyzing reaction products by X-ray diffraction and scanning transmission electron microscopy with energy dispersive spectroscopy. Geochemical simulations of aqueous systems containing PO4 and La-containing materials predicted that PO4 removal occurs via precipitation of poorly soluble LaPO4(s). Results from batch experiments agreed with those obtained from geochemical simulations, and mineralogical characterization of the reaction products were consistent with PO4 removal occurring primarily by precipitation of LaPO4(s). Between pH 1.5 and 12.9, La2(CO3)3(s) selectively removed PO4 over other anions from different aqueous matrices, including treated wastewater. However, the rate of PO4 removal decreased with increasing solution pH. In comparison to other solids, such as La(OH)3(s), La2(CO3)3(s) exhibits a relatively low solubility, particularly under slightly acidic conditions. Consequently, release of La3+ into the environment can be minimized when La2(CO3)3(s) is deployed for PO4 sequestration.

Influence of natural organic matter and pH on phosphate removal by and filterable lanthanum release from lanthanum-modified bentonite.

Lanthanum modified bentonite (LMB) has been applied to eutrophic lakes to reduce phosphorus (P) concentrations in the water column and mitigate P release from sediments. Previous experiments suggest that natural organic matter (NOM) can interfere with phosphate (PO4)-binding to LMB and exacerbate lanthanum (La)-release from bentonite. This evidence served as motivation for this study to systematically determine the effects of NOM, solution pH, and bentonite as a La carrier on P removal. We conducted both geochemical modeling and controlled-laboratory batch kinetic experiments to understand the pH-dependent impacts of humic and fulvic acids on PO4-binding to LMB and La release from LMB. The role of bentonite was studied by comparing PO4 removal obtained by LMB and La3+ (added as LaCl3 salt to represent the La-containing component of LMB). Our results from both geochemical modeling and batch experiments indicate that the PO4-binding ability of LMB is decreased in the presence of NOM, and the decrease is more pronounced at pH 8.5 than at 6. At the highest evaluated NOM concentration (28 mg C L-1), PO4-removal by La3+ was substantially lower than that by LMB, implying that bentonite clay in LMB shielded La from interactions with NOM, while still allowing PO4 capture by La. Finally, the presence of NOM promoted La-release from LMB, and the amount of La released depended on solution pH and both the type (i.e., fulvic/humic acid ratio) and concentration of NOM. Overall, these results provide an important basis for management of P in lakes and eutrophication control that relies on LMB applications.

Developing microbial communities containing a high abundance of exoelectrogenic microorganisms using activated carbon granules.

Microorganisms that can transfer electrons outside their cells are useful in a range of wastewater treatment and remediation technologies. Conventional methods of enriching exoelectrogens are cost-prohibitive (e.g., controlled-potential electrodes) or lack specificity (e.g., soluble electron acceptors). In this study a low-cost and simple approach to enrich exoelectrogens from a mixed microbial inoculum was investigated. After the method was validated using the exoelectrogen Geobacter sulfurreducens, microorganisms from a pilot-scale biological activated carbon (BAC) filter were subjected to incubations in which acetate was provided as the electron donor and granular activated carbon (GAC) as the electron acceptor. The BAC-derived community oxidized acetate and reduced GAC at a capacity of 1.0 mmol e- (g GAC)-1. After three transfers to new bottles, acetate oxidation rates increased 4.3-fold, and microbial morphologies and GAC surface coverage became homogenous. Although present at <0.01% in the inoculum, Geobacter species were significantly enriched in the incubations (up to 96% abundance), suggesting they were responsible for reducing the GAC. The ability to quickly and effectively develop an exoelectrogenic microbial community using GAC may help initiate and/or maintain environmental systems that benefit from the unique metabolic capabilities of these microorganisms.

Evidence of thermophilic waste decomposition at a landfill exhibiting elevated temperature regions.

There have been several reports of landfills exhibiting temperatures as high as 80 to 100 °C. This observation has motivated researchers to understand the causes of the elevated temperatures and to develop predictive models of landfill temperature. The objective of this research was to characterize the methanogenic activity of microbial communities that were derived from landfill samples excavated from a section of a landfill exhibiting gas well temperatures above 55 °C. Specific objectives were to: (1) determine the upper temperature limit for methane production; (2) evaluate the kinetics of methane generation when landfill-derived microcosms are incubated above and below their excavation temperature and derive a temperature inhibition function; and (3) evaluate microbial community shifts in response to temperature perturbations. Landfill microcosms were derived from 57 excavated landfill samples and incubated within ±2.5 °C of their excavation temperature between 42.5 °C and 87.5 °C. Results showed an optimum temperature for methane generation of ~57 °C and a 95% reduction in methane yield at ~72 °C. When select cultures were perturbed between 5 °C below and 15 °C above their in-situ temperature, both the rate and maximum methane production decreased as incubation temperature increased. Microbial community characterization using 16S rRNA amplicon sequencing suggests that thermophilic methanogenic activity can be attributed to methanogens of the genus Methanothermobacter. This study demonstrated that from a microbiological standpoint, landfills may maintain active methanogenic processes while experiencing temperatures in the thermophilic regime (<72 °C).

Emerging lanthanum (III)-containing materials for phosphate removal from water: A review towards future developments.

The last two decades have seen a rise in the development of lanthanum (III)-containing materials (LM) for controlling phosphate in the aquatic environment. >70 papers have been published on this topic in the peer-reviewed literature, but mechanisms of phosphate removal by LM as well as potential environmental impacts of LM remain unclear. In this review, we summarize peer-reviewed scientific articles on the development and use of 80 different types of LM in terms of prospective benefits, potential ecological impacts, and research needs. We find that the main benefits of LM for phosphate removal are their ability to strongly bind phosphate under diverse environmental conditions (e.g., over a wide pH range, in the presence of diverse aqueous constituents). The maximum phosphate uptake capacity of LM correlates primarily with the La content of LM, whereas reaction kinetics are influenced by LM formulation and ambient environmental conditions (e.g., pH, presence of co-existing ions, ligands, organic matter). Increased La solubilization can occur under some environmental conditions, including at moderately acidic pH values (i.e., < 4.5-5.6), highly saline conditions, and in the presence of organic matter. At the same time, dissolved La will likely undergo hydrolysis, bind to organic matter, and combine with phosphate to precipitate rhabdophane (LaPO4·H2O), all of which reduce the bioavailability of La in aquatic environments. Overall, LM use presents a low risk of adverse effects in water with pH > 7 and moderate-to-high bicarbonate alkalinity, although caution should be applied when considering LM use in aquatic systems with acidic pH values and low bicarbonate alkalinity. Moving forward, we recommend additional research dedicated to understanding La release from LM under diverse environmental conditions as well as long-term exposures on ecological organisms, particularly primary producers and benthic organisms. Further, site-specific monitoring could be useful for evaluating potential impacts of LM on both biotic and abiotic systems post-application.

Asymmetrical removal of sodium and chloride in flow-through capacitive deionization.

Capacitive deionization (CDI) is an electrochemical method of removing salt ions from brackish water. A common assumption in CDI is that monovalent ions (e.g., Na+, Cl-) are removed in a 1:1 symmetry on the electrodes. Validation of this assumption with techniques such as ion chromatography is not commonly performed, but is important to better understand how parasitic process, such as faradaic reactions, affect ion removals. In this study, we quantified the removals of Na+ and Cl- as a function of electrode orientation in flow-through CDI. When the cathode was positioned upstream, Na+ and Cl- removals approached a 1:1 symmetry, but when the anode was located upstream, we observed a significant drop in Na+, but not Cl-, removals. We attributed this drop to oxygen reduction reactions at the cathode that competed with Na+ adsorption. Oxidation of carbon in the upstream anode yielded H+ that enhanced the reduction of oxygen to H2O2 at the downstream cathode, which in turn diverted electrons from Na+ adsorption. In the absence of oxygen, Na+ removals increased in the upstream anode orientation and were comparable to Cl- removals, confirming that competition with oxygen reduction reactions was the primary reason for decreased Na+ removal. In the upstream cathode orientation, we show that H2O2 generated at the cathode can be oxidized at the downstream anode, possibly enhancing Na+ removals via internal electron recycling. Salt adsorption capacities calculated using actual ion removals did not always agree with those estimated using changes in solution conductivity, with the largest disagreement observed when conductivity data were corrected for pH changes. Our results highlight that faradaic reactions, particularly oxygen reduction reactions, can contribute to asymmetrical removals of monovalent ions in flow-through CDI.

Junction Potentials Bias Measurements of Ion Exchange Membrane Permselectivity.

Ion exchange membranes (IEMs) are versatile materials relevant to a variety of water and waste treatment, energy production, and industrial separation processes. The defining characteristic of IEMs is their ability to selectively allow positive or negative ions to permeate, which is referred to as permselectivity. Measured values of permselectivity that equal unity (corresponding to a perfectly selective membrane) or exceed unity (theoretically impossible) have been reported for cation exchange membranes (CEMs). Such nonphysical results call into question our ability to correctly measure this crucial membrane property. Because weighing errors, temperature, and measurement uncertainty have been shown to not explain these anomalous permselectivity results, we hypothesized that a possible explanation are junction potentials that occur at the tips of reference electrodes. In this work, we tested this hypothesis by comparing permselectivity values obtained from bare Ag/AgCl wire electrodes (which have no junction) to values obtained from single-junction reference electrodes containing two different electrolytes. We show that permselectivity values obtained using reference electrodes with junctions were greater than unity for CEMs. In contrast, electrodes without junctions always produced permselectivities lower than unity. Electrodes with junctions also resulted in artificially low permselectivity values for AEMs compared to electrodes without junctions. Thus, we conclude that junctions in reference electrodes introduce two biases into results in the IEM literature: (i) permselectivity values larger than unity for CEMs and (ii) lower permselectivity values for AEMs compared to those for CEMs. These biases can be avoided by using electrodes without a junction.

Hardwiring microbes via direct interspecies electron transfer: mechanisms and applications.

Multicellular microbial communities are important catalysts in engineered systems designed to treat wastewater, remediate contaminated sediments, and produce energy from biomass. Understanding the interspecies interactions within them is therefore essential to design effective processes. The flow of electrons within these communities is especially important in the determination of reaction possibilities (thermodynamics) and rates (kinetics). Conventional models of electron transfer incorporate the diffusion of metabolites generated by one organism and consumed by a second, frequently referred to as mediated interspecies electron transfer (MIET). Evidence has emerged in the last decade that another method, called direct interspecies electron transfer (DIET), may occur between organisms or in conjunction with electrically conductive materials. Recent research has suggested that DIET can be stimulated in engineered systems to improve desired treatment goals and energy recovery in systems such as anaerobic digesters and microbial electrochemical technologies. In this review, we summarize the latest understanding of DIET mechanisms, the associated microorganisms, and the underlying thermodynamics. We also critically examine approaches to stimulate DIET in engineered systems and assess their effectiveness. We find that in most cases attempts to promote DIET in mixed culture systems do not yield the improvements expected based on defined culture studies. Uncertainties of other processes that may be co-occurring in real systems, such as contaminant sorption and biofilm promotion, need to be further investigated. We conclude by identifying areas of future research related to DIET and its application in biological treatment processes.

Substrate and electrode potential affect electrotrophic activity of inverted bioanodes.

Electricity-consuming microbial communities can serve as biocathodic catalysts in microbial electrochemical technologies. Initiating their functionality, however, remains a challenge. One promising approach is the polarity inversion of bioanodes. The objective of this study was to examine the impact of bioanode substrate and electrode potentials on inverted electrotrophic activity. Bioanodes derived from domestic wastewater were operated at -0.15V or +0.15V (vs. standard hydrogen electrode) with either acetate or formate as the sole carbon source. After this enrichment phase, cathodic linear sweep voltammetry and polarization revealed that formate-enriched cultures consumed almost 20 times the current (-3.0±0.78mA; -100±26A/m(3)) than those established with acetate (-0.16±0.09mA; -5.2±2.9A/m(3)). The enrichment electrode potential had an appreciable impact for formate, but not acetate, adapted cultures, with the +0.15V enrichment generating twice the cathodic current of the -0.15V enrichment. The total charge consumed during cathodic polarization was comparable to the charge released during subsequent anodic polarization for the formate-adapted cultures, suggesting that these communities accumulated charge or generated reduced products that could be rapidly oxidized. These findings imply that it may be possible to optimize electrotrophic activity through specific bioanodic enrichment procedures.

Correction to Comparison of Nonprecious Metal Cathode Materials for Methane Production by Electromethanogenesis.

[This corrects the article DOI: 10.1021/sc400520x.].

Comparison of Nonprecious Metal Cathode Materials for Methane Production by Electromethanogenesis.

In methanogenic microbial electrolysis cells (MMCs), CO2 is reduced to methane using a methanogenic biofilm on the cathode by either direct electron transfer or evolved hydrogen. To optimize methane generation, we examined several cathode materials: plain graphite blocks, graphite blocks coated with carbon black or carbon black containing metals (platinum, stainless steel or nickel) or insoluble minerals (ferrihydrite, magnetite, iron sulfide, or molybdenum disulfide), and carbon fiber brushes. Assuming a stoichiometric ratio of hydrogen (abiotic):methane (biotic) of 4:1, methane production with platinum could be explained solely by hydrogen production. For most other materials, however, abiotic hydrogen production rates were insufficient to explain methane production. At -600 mV, platinum on carbon black had the highest abiotic hydrogen gas formation rate (1600 ± 200 nmol cm-3 d-1) and the highest biotic methane production rate (250 ± 90 nmol cm-3 d-1). At -550 mV, plain graphite (76 nmol cm-3 d-1) performed similarly to platinum (73 nmol cm-3 d-1). Coulombic recoveries, based on the measured current and evolved gas, were initially greater than 100% for all materials except platinum, suggesting that cathodic corrosion also contributed to electromethanogenic gas production.

Improving startup performance with carbon mesh anodes in separator electrode assembly microbial fuel cells.

In a separator electrode assembly microbial fuel cell, oxygen crossover from the cathode inhibits current generation by exoelectrogenic bacteria, resulting in poor reactor startup and performance. To determine the best approach for improving startup performance, the effect of acclimation to a low set potential (-0.2V, versus standard hydrogen electrode) was compared to startup at a higher potential (+0.2 V) or no set potential, and inoculation with wastewater or pre-acclimated cultures. Anodes acclimated to -0.2 V produced the highest power of 1330±60 mW m(-2) for these different anode conditions, but unacclimated wastewater inocula produced inconsistent results despite the use of this set potential. By inoculating reactors with transferred cell suspensions, however, startup time was reduced and high power was consistently produced. These results show that pre-acclimation at -0.2 V consistently improves power production compared to use of a more positive potential or the lack of a set potential.

Enrichment of microbial electrolysis cell biocathodes from sediment microbial fuel cell bioanodes.

Electron-accepting (electrotrophic) biocathodes were produced by first enriching graphite fiber brush electrodes as the anodes in sediment-type microbial fuel cells (sMFCs) using two different marine sediments and then electrically inverting the anodes to function as cathodes in two-chamber bioelectrochemical systems (BESs). Electron consumption occurred at set potentials of -439 mV and -539 mV (versus the potential of a standard hydrogen electrode) but not at -339 mV in minimal media lacking organic sources of energy. Results at these different potentials were consistent with separate linear sweep voltammetry (LSV) scans that indicated enhanced activity (current consumption) below only ca. -400 mV. MFC bioanodes not originally acclimated at a set potential produced electron-accepting (electrotrophic) biocathodes, but bioanodes operated at a set potential (+11 mV) did not. CO(2) was removed from cathode headspace, indicating that the electrotrophic biocathodes were autotrophic. Hydrogen gas generation, followed by loss of hydrogen gas and methane production in one sample, suggested hydrogenotrophic methanogenesis. There was abundant microbial growth in the biocathode chamber, as evidenced by an increase in turbidity and the presence of microorganisms on the cathode surface. Clone library analysis of 16S rRNA genes indicated prominent sequences most similar to those of Eubacterium limosum (Butyribacterium methylotrophicum), Desulfovibrio sp. A2, Rhodococcus opacus, and Gemmata obscuriglobus. Transfer of the suspension to sterile cathodes made of graphite plates, carbon rods, or carbon brushes in new BESs resulted in enhanced current after 4 days, demonstrating growth by these microbial communities on a variety of cathode substrates. This report provides a simple and effective method for enriching autotrophic electrotrophs by the use of sMFCs without the need for set potentials, followed by the use of potentials more negative than -400 mV.

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.

Syntrophic interactions improve power production in formic acid fed MFCs operated with set anode potentials or fixed resistances.

Formic acid is a highly energetic electron donor but it has previously resulted in low power densities in microbial fuel cells (MFCs). Three different set anode potentials (-0.30, -0.15, and +0.15 V; vs. a standard hydrogen electrode, SHE) were used to evaluate syntrophic interactions in bacterial communities for formic acid degradation relative to a non-controlled, high resistance system (1,000 Ω external resistance). No current was generated at -0.30 V, suggesting a lack of direct formic acid oxidation (standard reduction potential: -0.40 V). More positive potentials that allowed for acetic acid utilization all produced current, with the best performance at -0.15 V. The anode community in the -0.15 V reactor, based on 16S rDNA clone libraries, was 58% Geobacter sulfurreducens and 17% Acetobacterium, with lower proportions of these genera found in the other two MFCs. Acetic acid was detected in all MFCs suggesting that current generation by G. sulfurreducens was dependent on acetic acid production by Acetobacterium. When all MFCs were subsequently operated at an external resistance for maximum power production (100 Ω for MFCs originally set at -0.15 and +0.15 V; 150 Ω for the control), they produced similar power densities and exhibited the same midpoint potential of -0.15 V in first derivative cyclic voltammetry scans. All of the mixed communities converged to similar proportions of the two predominant genera (ca. 52% G. sulfurreducens and 22% Acetobacterium). These results show that syntrophic interactions can be enhanced through setting certain anode potentials, and that long-term performance produces stable and convergent communities.

Lactate oxidation coupled to iron or electrode reduction by Geobacter sulfurreducens PCA.

Geobacter sulfurreducens PCA completely oxidized lactate and reduced iron or an electrode, producing pyruvate and acetate intermediates. Compared to the current produced by Shewanella oneidensis MR-1, G. sulfurreducens PCA produced 10-times-higher current levels in lactate-fed microbial electrolysis cells. The kinetic and comparative analyses reported here suggest a prominent role of G. sulfurreducens strains in metal- and electrode-reducing communities supplied with lactate.

Adaptation to high current using low external resistances eliminates power overshoot in microbial fuel cells.

One form of power overshoot commonly observed with mixed culture microbial fuel cells (MFCs) is doubling back of the power density curve at higher current densities, but the reasons for this type of overshoot have not been well explored. To investigate this, MFCs were acclimated to different external resistances, producing a range of anode potentials and current densities. Power overshoot was observed for reactors acclimated to higher (500 and 5000 Ω) but not lower (5 and 50 Ω) resistances. Acclimation of the high external resistance reactors for a few cycles to low external resistance (5 Ω), and therefore higher current densities, eliminated power overshoot. MFCs initially acclimated to low external resistances exhibited both higher current in cyclic voltammograms (CVs) and higher levels of redox activity over a broader range of anode potentials (-0.4 to 0 V; vs. a Ag/AgCl electrode) based on first derivative cyclic voltammetry (DCV) plots. Reactors acclimated to higher external resistances produced lower current in CVs, exhibited lower redox activity over a narrower anode potential range (-0.4 to -0.2 V vs. Ag/AgCl), and failed to produce higher currents above ∼-0.3 V (vs. Ag/AgCl). After the higher resistance reactors were acclimated to the lowest resistance they also exhibited similar CV and DCV profiles. Our findings show that to avoid overshoot, prior to the polarization and power density tests the anode biofilm must adapt to low external resistances to be capable of higher currents.

A method for high throughput bioelectrochemical research based on small scale microbial electrolysis cells.

There is great interest in studying exoelectrogenic microorganisms, but existing methods can require expensive electrochemical equipment and specialized reactors. We developed a simple system for conducting high throughput bioelectrochemical research using multiple inexpensive microbial electrolysis cells (MECs) built with commercially available materials and operated using a single power source. MECs were small crimp top serum bottles (5 mL) with a graphite plate anode (92 m²/m(3)) and a cathode of stainless steel (SS) mesh (86 m²/m³), graphite plate, SS wire, or platinum wire. The highest volumetric current density (240 A/m³, applied potential of 0.7 V) was obtained using a SS mesh cathode and a wastewater inoculum (acetate electron donor). Parallel operated MECs (single power source) did not lead to differences in performance compared to non-parallel operated MECs, which can allow for high throughput reactor operation (>1000 reactors) using a single power supply. The utility of this method for cultivating exoelectrogenic microorganisms was demonstrated through comparison of buffer effects on pure (Geobacter sulfurreducens and Geobacter metallireducens) and mixed cultures. Mixed cultures produced current densities equal to or higher than pure cultures in the different media, and current densities for all cultures were higher using a 50 mM phosphate buffer than a 30 mM bicarbonate buffer. Only the mixed culture was capable of sustained current generation with a 200 mM phosphate buffer. These results demonstrate the usefulness of this inexpensive method for conducting in-depth examinations of pure and mixed exoelectrogenic cultures.