Flow-through laminar anodes with variable interlaminar distance to modulate the current density of urine-fed bio-electrochemical systems.

Three-dimensional (3D) porous anodes used in urine-powered bio-electrochemical applications usually lead to the growth of electro-active bacteria on the outer electrode surface, due to limited microbial access to the internal structure and lack of permeation of culture medium through the entire porous architecture. In this study, we propose the use of 3D monolithic Ti4O7 porous electrodes with controlled laminar structures as microbial anodes for urine-fed bio-electrochemical systems. The interlaminar distance was tuned to modulate the anode surface areas and, thus, the volumetric current densities. To profit from the true area of the electrodes, urine feeding was performed as a continuous flow through the laminar architectures. The system was optimized according to the response surface methodology (RSM). The electrode interlaminar distance and the concentration of urine were selected as independent variables, with the volumetric current density as the output response to optimize. Maximum current densities of 5.2 kA.m-3 were produced from electrodes with 12 µm-interlaminar distance and 10 %v/v urine concentrations. The present study demonstrates the existence of a trade-off between the accesibility to the internal electrode structure and the effective usage of the surface area to maximize the volumetric current density when diluted urine is used as flowing-through feeding fuel.

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), Rext 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 (kDN ) 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.

Engineering Wired Life: Synthetic Biology for Electroactive Bacteria.

Electroactive bacteria produce or consume electrical current by moving electrons to and from extracellular acceptors and donors. This specialized process, known as extracellular electron transfer, relies on pathways composed of redox active proteins and biomolecules and has enabled technologies ranging from harvesting energy on the sea floor, to chemical sensing, to carbon capture. Harnessing and controlling extracellular electron transfer pathways using bioengineering and synthetic biology promises to heighten the limits of established technologies and open doors to new possibilities. In this review, we provide an overview of recent advancements in genetic tools for manipulating native electroactive bacteria to control extracellular electron transfer. After reviewing electron transfer pathways in natively electroactive organisms, we examine lessons learned from the introduction of extracellular electron transfer pathways into Escherichia coli. We conclude by presenting challenges to future efforts and give examples of opportunities to bioengineer microbes for electrochemical applications.

Bioelectrical Methane Production with an Ammonium Oxidative Reaction under the No Organic Substance Condition.

The present study investigated bioelectrical methane production from CO2 without organic substances. Even though microbial methane production has been reported at relatively high electric voltages, the amount of voltage required and the organisms contributing to the process currently remain unknown. Methane production using a biocathode was investigated in a microbial electrolysis cell coupled with an NH4+ oxidative reaction at an anode coated with platinum powder under a wide range of applied voltages and anaerobic conditions. A microbial community analysis revealed that methane production simultaneously occurred with biological denitrification at the biocathode. During denitrification, NO3- was produced by chemical NH4+ oxidation at the anode and was provided to the biocathode chamber. H2 was produced at the biocathode by the hydrogen-producing bacteria Petrimonas through the acceptance of electrons and protons. The H2 produced was biologically consumed by hydrogenotrophic methanogens of Methanobacterium and Methanobrevibacter with CO2 uptake and by hydrogenotrophic denitrifiers of Azonexus. This microbial community suggests that methane is indirectly produced without the use of electrons by methanogens. Furthermore, bioelectrical methane production occurred under experimental conditions even at a very low voltage of 0.05| |V coupled with NH4+ oxidation, which was thermodynamically feasible.

Study on the effect of synergy effect between the mixed cultures on the power generation of microbial fuel cells.

Microbial fuel cells (MFC) can use microorganisms to directly convert the chemical energy of organic matter into electrical energy, and generate electrical energy while pollutants degradation. To solve the critical problem of lower power yield of power production, this study selected Saccharomyces cerevisiae, Escherichia coli, Pseudomonas aeruginosa, and Bacillus subtilis as the anodic inoculums. The influence of the mixed bacteria on the power-producing effect of MFC and the synergy effect between the electrochemically active bacteria in mixed cultures were discussed. The results showed that among the mixed culture system, only the mixed cultures MFC composed of Saccharomyces cerevisiae and Bacillus subtilis had a significant increase in power generation capacity, which could reach to 554 mV. Further analysis of the electrochemical and microbiological performance of this system was conducted afterward to verify the synergy effect between Saccharomyces cerevisiae and Bacillus subtilis. The riboflavin produced by Bacillus subtilis could be utilized by Saccharomyces cerevisiae to enhance the power generation capacity. Meanwhile, Saccharomyces cerevisiae could provide carbon source and electron donor for Bacillus subtilis through respiration. Finally, in the experiment of adding exogenous riboflavin in the mixed bacterial MFC, the result indicated that the mixed bacterial MFC chose the self-secreting riboflavin over the exogenous riboflavin as the electron mediator, and the excess riboflavin might hinder the electron trasfer.

Long-distance electron transfer in a filamentous Gram-positive bacterium.

Long-distance extracellular electron transfer has been observed in Gram-negative bacteria and plays roles in both natural and engineering processes. The electron transfer can be mediated by conductive protein appendages (in short unicellular bacteria such as Geobacter species) or by conductive cell envelopes (in filamentous multicellular cable bacteria). Here we show that Lysinibacillus varians GY32, a filamentous unicellular Gram-positive bacterium, is capable of bidirectional extracellular electron transfer. In microbial fuel cells, L. varians can form centimetre-range conductive cellular networks and, when grown on graphite electrodes, the cells can reach a remarkable length of 1.08 mm. Atomic force microscopy and microelectrode analyses suggest that the conductivity is linked to pili-like protein appendages. Our results show that long-distance electron transfer is not limited to Gram-negative bacteria.

Identification of a Diguanylate Cyclase That Facilitates Biofilm Formation on Electrodes by Shewanella oneidensis MR-1.

In many bacteria, cyclic diguanosine monophosphate (c-di-GMP), synthesized by diguanylate cyclase (DGC), serves as a second messenger involved in the regulation of biofilm formation. Although studies have suggested that c-di-GMP also regulates the formation of electrochemically active biofilms (EABFs) by Shewanella oneidensis MR-1, DGCs involved in this process remained to be identified. Here, we report that the SO_1646 gene, hereafter named dgcS, is upregulated under medium flow conditions in electrochemical flow cells (EFCs), and its product (DgcS) functions as a major DGC in MR-1. In vitro assays demonstrated that purified DgcS catalyzed the synthesis of c-di-GMP from GTP. Comparisons of intracellular c-di-GMP levels in the wild-type strain and a dgcS deletion mutant (ΔdgcS mutant) showed that production of c-di-GMP was markedly reduced in the ΔdgcS mutant when cells were grown in batch cultures and on electrodes in EFCs. Cultivation of the ΔdgcS mutant in EFCs also revealed that the loss of DgcS resulted in impaired biofilm formation and decreased current generation. These findings demonstrate that MR-1 uses DgcS to synthesize c-di-GMP under medium flow conditions, thereby activating biofilm formation on electrodes.IMPORTANCE Bioelectrochemical systems (BESs) have attracted wide attention owing to their utility in sustainable biotechnology processes, such as microbial fuel cells and electrofermentation systems. In BESs, electrochemically active bacteria (EAB) form biofilms on electrode surfaces, thereby serving as effective catalysts for the interconversion between chemical and electric energy. It is therefore important to understand mechanisms for the formation of biofilm by EAB grown on electrodes. Here, we show that a model EAB, S. oneidensis MR-1, expresses DgcS as a major DGC, thereby activating the formation of biofilms on electrodes via c-di-GMP-dependent signal transduction cascades. The findings presented herein provide the molecular basis for improving electrochemical interactions between EAB and electrodes in BESs. The results also offer molecular insights into how Shewanella regulates biofilm formation on solid surfaces in the natural environment.

Bacterial strains colonizing the sensor electrodes of a continuous glucose monitoring system in children with diabetes.

INTRODUCTION: The higher frequency of infections in diabetic patients is caused by a hyperglycemic environment, which promotes immune dysfunction. People with diabetes are more prone to skin infections. A continuous glucose monitoring (CGM) system provides information on changes in blood glucose (BG) levels throughout the day. Its use facilitates optimal therapeutic decisions for a diabetic patient. One of the factors limiting the use of CGM is inflammation at the insertion site. AIM OF THE STUDY: The aim of the study was the microbiological identification of the bacterial strains which are found on CGM sensor electrodes. MATERIAL AND METHODS: We performed microbiological tests on patients' CGM Enlite Medtronic electrodes, which were removed after 6 days of usage according to the manufacturer's instructions. 31 sensors were examined from 31 children (14 girls) aged from 0.5 to 14.6 years. The microbiological analysis was routinely performed at the Department of Children's Diabetology Medical University of Silesia in Katowice, Poland. RESULTS: 12 (39%) of the electrodes were colonized. In 11 (92%) cases the electrodes were colonized by one bacteria strain. 7 times methicillin-sensitive coagulase negative staphylococcus (MSCNS) was detected. We also found one case of Klebsiella pneumoniae, Ochrobactrum tritici, Bacillus sonorensis and methicillin-resistant coagulase-negative Staphylococci (MRCNS) colonization. One electrode was colonized by the mixed flora Enterococcus faecalis, methicillin-susceptible coagulase-negative Staphylococci (MSCNS), Pseudomonas stutzeri, methicillin-susceptible Staphylococcus aureus (MSSA). The median HbA1c in the group with colonization of electrodes was 6, 85% (6, 3-7, 6%) versus 6, 3% (5, 8-7, 5%) in the group without colonization. The median BMI in the group with colonization of the electrodes was 17.10 kg/m2 (16.28-18.62 kg/m2) versus 15.98 kg/m2 (15.14-17.96 kg/m2) in the group without colonization. Statistically, significantly more frequently electrodes are colonized in older children (median age in the group with colonization of electrodes 11.43 years (6.52-12.27 years), without colonization 8.42 years. (3.098-9.375 years); (p = 0.033). CONCLUSIONS: It seems that older children are more likely to have their sensor electrode colonized by bacterial strains.

Electrolocation? The evidence for redox-mediated taxis in Shewanella oneidensis.

Shewanella oneidensis is a dissimilatory metal reducing bacterium and model for extracellular electron transfer (EET), a respiratory mechanism in which electrons are transferred out of the cell. In the last 10 years, migration to insoluble electron acceptors for EET has been shown to be nonrandom and tactic, seemingly in the absence of molecular or energy gradients that typically allow for taxis. As the ability to sense, locate, and respire electrodes has applications in bioelectrochemical technology, a better understanding of taxis in S. oneidensis is needed. While the EET conduits of S. oneidensis have been studied extensively, its taxis pathways and their interplay with EET are not yet understood, making investigation into taxis phenomena nontrivial. Since S. oneidensis is a member of an EET-encoding clade, the genetic circuitry of taxis to insoluble acceptors may be conserved. We performed a bioinformatic analysis of Shewanella genomes to identify S. oneidensis chemotaxis orthologs conserved in the genus. In addition to the previously reported core chemotaxis gene cluster, we identify several other conserved proteins in the taxis signaling pathway. We present the current evidence for the two proposed models of EET taxis, "electrokinesis" and flavin-mediated taxis, and highlight key areas in need of further investigation.

Plant secondary metabolites induced electron flux in microbial fuel cell: investigation from laboratory-to-field scale.

Wastewater treatment coupled with electricity recovery in microbial fuel cell (MFC) prefer mixed anaerobic sludge as inoculum in anodic chamber than pure stain of electroactive bacteria (EAB), due to robustness and syntrophic association. Genetic modification is difficult to adopt for mixed sludge microbes for enhancing power production of MFC. Hence, we demonstrated use of eco-friendly plant secondary metabolites (PSM) with sub-lethal concentrations to enhance the rate of extracellular electron transfer between EAB and anode and validated it in both bench-scale as well as pilot-scale MFCs. The PSMs contain tannin, saponin and essential oils, which are having electron shuttling properties and their addition to microbes can cause alteration in cell morphology, electroactive behaviour and shifting in microbial population dynamics depending upon concentrations and types of PSM used. Improvement of 2.1-times and 3.8-times in power densities was observed in two different MFCs inoculated with Eucalyptus-extract pre-treated mixed anaerobic sludge and pure culture of Pseudomonas aeruginosa, respectively, as compared to respective control MFCs operated without adding Eucalyptus-extract to inoculum. When Eucalyptus-extract-dose was spiked to anodic chamber (125 l) of pilot-scale MFC, treating septage, the current production was dramatically improved. Thus, PSM-dosing to inoculum holds exciting promise for increasing electricity production of field-scale MFCs.

Bacterial response to formaldehyde in an MFC toxicity sensor.

Microbial fuel cells (MFCs) have been extensively studied as toxicity sensors. MFCs have potential as toxicity sensors because when their anodes are exposed to toxic substances or the concentration of the target is changed, the voltage generated by certain microorganisms with active electrical activity attached to the anode changes. However, the underlying reasons behind this phenomenon have not been deeply explored. Therefore, the activity of anodic microorganisms during voltage drops in MFCs under formaldehyde (FA) stress was studied. The composition of the microbial community structure with similar active voltage drops under different concentrations of FA was investigated. The results showed that under exposure to high concentrations (169.20 mg/L) of FA, the voltage changes in MFCs could be divided into five stages. With prolonged exposure to FA, the ratio of live/dead bacteria decreased from 4.78 to 0.65. This result indicated that voltage drops may be caused by FA causing cell membrane rupture. Analysis of the microbial community structure under treatments including inoculation and three concentrations of FA revealed that Geobacter and other electrogenic bacteria were effectively enriched in the MFCs. Low-concentration and long-term exposure to FA had a greater impact on microbial communities than high-concentration and short-term exposure. Pseudomonas and Acidovorax were more significantly affected by FA than Flavobacterium and Geobacter. This study explains the reason for the voltage drop of MFCs after exposure to toxic substances from the perspective of the microorganisms.

Direct cathodic electron uptake coupled to sulfate reduction by Desulfovibrio ferrophilus IS5 biofilms.

Direct electron uptake is emerging as a key process for electron transfer in anaerobic microbial communities, both between species and from extracellular sources, such as zero-valent iron (Fe0 ) or cathodic surfaces. In this study, we investigated cathodic electron uptake by Fe0 -corroding Desulfovibrio ferrophilus IS5 and showed that electron uptake is dependent on direct cell contact via a biofilm on the cathode surface rather than through secreted intermediates. Induction of cathodic electron uptake by lactate-starved D. ferrophilus IS5 cells resulted in the expression of all components necessary for electron uptake; however, protein synthesis was required for full biofilm formation. Notably, proteinase K treatment uncoupled electron uptake from biofilm formation, likely through proteolytic degradation of proteinaceous components of the electron uptake machinery. We also showed that cathodic electron uptake is dependent on SO4 2- reduction. The insensitivity of Fe0 corrosion to proteinase K treatment suggests that electron uptake from a cathode might involve different mechanism(s) than those involved in Fe0 corrosion.

Development of an electroactive aerobic biocathode for microbial fuel cell applications.

Microbial biocathodes are gaining interest due to their low cost, environmental friendliness and sustainable nature. In this study, a microbial consortium was enriched from activated sludge obtained from a common textile effluent treatment plant in the absence of organic carbon source to produce an electroactive biofilm. Chronoamperometry method of enrichment was carried out for over 70 days to select for electroactive bacteria that could be used as a cathode catalyst in microbial fuel cells (MFC). The resultant biofilm produced an average peak current of -0.7 mA during the enrichment and produced a maximum power density of 64.6 ± 3.5 mW m-2 compared to platinum (72.7 ± 1.2 mW m-2 ) in a Shewanella-based MFC. Microbial community analysis of the initial sludge sample and enriched samples, based on 16S rRNA gene sequencing, revealed the selection of chemolithotrophs with the most dominant phylum being Bacteroidetes, Proteobacteria, Firmicutes, Actinobacteria and Acidobacteria in the enriched samples. A variety of CO2 fixing and nitrate-reducing bacteria was present in the resultant biofilm on the cathode. This study suggests that microbial consortia are capable of replacing expensive platinum as a cathode catalyst in MFCs.

Influence of long-term organic and conventional fertilization on bacterial communities involved in bioelectricity production from paddy field-microbial fuel cells.

Soils are rich in organics, particularly those that support the growth of plants. These organics are possible sources of sustainable energy, and a microbial fuel cell (MFC) system can potentially be used for this purpose. In this, the soil organic content expelled from plant root was possibly converted into electrical energy through the microbial metabolic process. The integration of MFC systems with living plant root system is a novel approach, which will facilitate sustainable resource for energy production. Therefore, the objective of this study is to electrochemically evaluate the paddy field MFCs (PF-MFCs) performance and methane emission under organic and conventional fertilization systems in paddy fields, and its impact on bacterial communities involved in bioelectricity production. Graphite (anode) and carbon (cathode) electrode MFC systems were configured and assembled in organic and conventionally fertilized paddy fields. The anode and bulk soil-associated bacterial communities were examined using high-throughput Illumina MiSeq sequencing platform. Our results revealed that the maximum electricity production and power density were observed in CFPF-MFC with less methane emission compared to OFPF-MFC. The next-generation sequencing (NGS) libraries showed that the bacterial population was significantly increased in the organic-fertilized field and the enhanced occurrence of the Geobacteraceae family in CFPF-MFC anode. By enhancing Geobacteraceae occurrence on the anode, the conventional fertilization improved the bioelectricity production with less methane emission. Further extension in the establishment of plant MFCs in various sedimentary environments will solve the global energy crisis.

Active N dopant states of electrodes regulate extracellular electron transfer of Shewanella oneidensis MR-1 for bioelectricity generation: Experimental and theoretical investigations.

Anodic N doping is an effective way to improve power generation of bioelectrochemical systems (BESs), but the role of various active N dopant states of the anode on BES performance is still unclear. Herein, the effect of anodic active N dopant states on bioelectricity generation of Shewanella oneidensis MR-1 inoculated BESs particularly including microbial extracellular electron transfer (EET) was explored using experiments and theoretical simulations. It was found a positive linear correlation between the peak current density of BESs and pyrrolic N content of the anode, which would mainly ascribe to the enhancement of both direct electron transfer (DET) and mediated electron transfer (MET) of S. oneidensis MR-1. Morever, the molecule dynamic simulation revealed that such EET improvements of S. oneidensis MR-1 could be due to more remarkable reduction in the thermodynamic and kinetic resistances of the DET and MET processes with anodic doping of pyrrolic N compared to pyridinic N and graphitic N. This work provides a valuable guideline to design of high-performance anodes for potential BES applications.

In-situ mineral CO2 sequestration in a methane producing microbial electrolysis cell treating sludge hydrolysate.

Microbial electrolysis cell (MEC) has excellent CH4 production performance, however, CO2 still remains in the produced biogas at high content. For achieving in-situ CO2 sequestration and thus upgrading biogas, mineral carbonation was integrated into a MEC treating sludge hydrolysate. With 19 g/L wollastonite addition, in-situ mineral CO2 sequestration was achieved by formation of calcite precipitates. CH4 content in the biogas was increased by 5.1 % and reached 95.9 %, with CH4 production improved by 16.9 %. In addition, the removals of polysaccharide, protein, and chemical oxygen demand (COD) of the MEC were increased by 4.4 %, 6.7 %, and 8.4 %, respectively. The generated precipitates rarely accumulated on bio-cathode, and did not significantly affect the morphology of cathode biofilm. However, integrating mineral carbonation resulted in a higher relative abundance of Methanosarcina on anode and slightly decreased the ratio of Methanobacterium to Methanosaeta on cathode, which should be noticed. In conclusion, integrating mineral carbonation is an attractive way to improve the performance of MEC by achieving in-situ CO2 sequestration, accompanied with CH4 production enhancement.

Mutual effect between electrochemically active bacteria (EAB) and azo dye in bio-electrochemical system (BES).

Herein, the mutual effect between azo dye and the performance of electrochemically active bacteria (EAB) is investigated in detail, which is crucial to understand and control the bio-electrochemical systems (BESs) operation for azo dye containing wastewater treatment. EAB is enriched at controlled potential of -0.2 V vs Ag/AgCl in single-chamber BESs. Over 95% azo dye (alizarin yellow R (AYR)) was decolorized regardless of the initial AYR concentration ranging from 30 to 120 mg/L within 24 h. The fastest decolorization rate was obtained at AYR initial concentration of 70 mg/L, which was 4.25 times greater in the closed circuit BESs than that in the open circuit one. 16S rRNA gene based microbial community analysis showed that Geobacter was dominant in EAB with relative abundance increased from 77.98% (0 mg/L AYR) to 92.22% (70 mg/L AYR), indicating that azo dye selectively boosts the growth of exoelectrogens in electrode biofilm communities. Under electricity stimulation, extracellular process can be mutually conducted by azo dye compounds, which is favorable for accelerating reaction rate and avoiding of significant toxic effect on EAB.

Production of Current by Syntrophy Between Exoelectrogenic and Fermentative Hyperthermophilic Microorganisms in Heterotrophic Biofilm from a Deep-Sea Hydrothermal Chimney.

To study the role of exoelectrogens within the trophic network of deep-sea hydrothermal vents, we performed successive subcultures of a hyperthermophilic community from a hydrothermal chimney sample on a mix of electron donors in a microbial fuel cell system. Electrode (the electron acceptor) was swapped every week to enable fresh development from spent media as inoculum. The MFC at 80 °C yielded maximum current production increasing from 159 to 247 mA m-2 over the subcultures. The experiments demonstrated direct production of electric current from acetate, pyruvate, and H2 and indirect production from yeast extract and peptone through the production of H2 and acetate from fermentation. The microorganisms found in on-electrode communities were mainly affiliated to exoelectrogenic Archaeoglobales and Thermococcales species, whereas in liquid media, the communities were mainly affiliated to fermentative Bacillales and Thermococcales species. The work shows interactions between fermentative microorganisms degrading complex organic matter into fermentation products that are then used by exoelectrogenic microorganisms oxidizing these reduced compounds while respiring on a conductive support. The results confirmed that with carbon cycling, the syntrophic relations between fermentative microorganisms and exoelectrogens could enable some microbes to survive as biofilm in extremely unstable conditions. Graphical Abstract Schematic representation of cross-feeding between fermentative and exoelectrogenic microbes on the surface of the conductive support. B, Bacillus/Geobacillus spp.; Tc, Thermococcales; Gg, Geoglobus spp.; Py, pyruvate; Ac, acetate.

Inducing laccase activity in white rot fungi using copper ions and improving the efficiency of azo dye treatment with electricity generation using microbial fuel cells.

This work presents a white rot fungus-microbial fuel cell (WRF-MFC) that uses WRF that is grown at its cathode. Adding Cu2+ to the fungi-containing solid medium stimulated WRF-secreting laccase, which catalyzed the redox reaction in the MFC and thereby promoting the generation of electricity. Adding 12.5 mg L-1 Cu2+ to a G. lucidum-containing medium provided the greatest laccase stimulation and increased the laccase activity by a factor of 1.6. Adding 12.5 mg L-1 Cu2+ to the WRF chamber of WRF-MFC increased its decolorization of Acid Orange 7 (AO-7) and increased its power density to 223 mW m-2, which was 1.77 times that of an MFC without WRF. The enhancement of decolorization and electricity generation improved the performance of the WRF-MFC, indicating that a laccase-catalyzed cathode has great potential effectiveness in microbial fuel cells.

Trophic networks improve the performance of microbial anodes treating wastewater.

Microbial anodes represent a distinct ecological niche that is characterized mainly by the terminal electron acceptor, i.e., the anode potential, and the substrate, i.e., the electron source. Here, we determine the performance and the biofilm community of anode microbiomes while using substrates of increasing complexity (organic acids or organic acids and sugar or real domestic wastewater) to mimic different, practically relevant, trophic levels. α-Diversity values increased with substrate complexity. In addition, the higher abundance value of Deltaproteobacteria in the biofilms corresponds to higher reactor performance (i.e., COD removal, current density, and Coulombic efficiency). In reactors exploiting real wastewater, the diversity of the planktonic microorganisms was only little affected. Microbiome network analysis revealed two important clusters for reactor performance as well as performance-independent pathogen-containing clusters. Interestingly, Geobacter was not found to be integrated in the network underlining its outstanding individual ecological role in line with its importance for the efficiency of the electron harvest for all reactors. The microbiome analysis of different trophic levels and their temporal development from initial colonization to stable treatment demonstrate important principles for the implementation of microbial anodes for wastewater treatment.