Anodic Biofilm Research Articles

Insight into electricity production performance from dried waste activated sludge in dual chamber microbial fuel cells: Influencing factors, neural network modelling and microbial community analysis

Directly deriving electrical energy from waste activated sludge (WAS) by using microbial fuel cell (MFC) is one of the most promising alternatives to resource utilization of WAS. However, obtaining high MFC performance is still a great challenge for the full-scale implementation of MFC. Therefore, this study aimed to quantify and model the effect of different operating parameters on the performance of MFC fed with dried WAS as anode substrate. The results indicated that MFC exhibited a fast start-up time of 3 days at the inoculated anaerobic sludge: dry WAS mass ratio of 1:5. Moreover, the output voltage and power density of MFC were highly dependent of anolyte pH and temperature, WAS concentration, which was well simulated and predicted by the BPANN model. It was revealed that MFC fed with dry WAS exhibited good performance at 55ºC, pH 9.0 and WAS concentration of 16 g/L, while the stable output voltage and power density were 0.705 V and 5.248 W/m3, respectively. The anolyte temperature significantly affected microbial communities of the anode biofilm. At the phylum level, Synergistota and Proteobacteria dominated the anode biofilm at 35ºC, while Firmicutes was absolutely enriched in the anode biofilm at 55ºC. At the genus level, the predominant Advenella is detected in the anode biofilm at 35ºC, whereas the preponderant MBA03 is identified in the anode biofilm at 35ºC. The findings herein may provide a route for efficiently driving power energy from WAS, which can facilitate the fulfillment of full-scale MFC for the resource utilization of WAS.

Fabrication and optimization of bioelectrochemical system using tetracycline-degrading bacterial strains for antibiotic wastewater treatment

In this study, a microbial fuel cell was constructed using Raoultella sp. XY-1 to efficiently degrade tetracycline (TC) and assess the effectiveness of the electrochemical system. The degradation rate reached 83.2 ± 1.8 % during the 7-day period, in which the system contained 30 mg/L TC, and the degradation pathway and intermediates were identified. Low concentrations of TC enhanced anodic biofilm power production, while high concentrations of TC decreased the electrochemical activity of the biofilm, extracellular polymeric substances, and enzymatic activities associated with electron transfer. Introducing electrogenic bacteria improved power generation efficiency. A three-strain hybrid system was fabricated using Castellaniella sp. A3, Castellaniella sp. A5 and Raoultella sp. XY-1, leading to the enhanced TC degradation rate of 90.4 % and the increased maximum output voltage from 200 to 265 mV. This study presents a strategy utilizing tetracycline-degrading bacteria as bioanodes for TC removal, while incorporating electrogenic bacteria to enhance electricity generation.

Resilience of anodic biofilm in microbial fuel cell biosensor for BOD monitoring of urban wastewater

Efficient wastewater treatment monitoring is vital for addressing water scarcity. Microbial fuel cells (MFCs) have emerged as real-time biosensors for biochemical oxygen demand (BOD) in urban wastewater. Discrepancies in signal generation may arise due to changes in the composition and metabolism of mixed-culture electroactive biofilms stemming from different wastewater compositions. In this study, 3D-printed MFC-based biosensors were employed to assess the BOD of sterile complex artificial wastewater and untreated urban wastewater. Alterations in the microbial composition of the anode were evaluated using 16S rRNA sequencing and metagenomics analysis. Results show that MFC-based biosensors can be effectively recalibrated for diverse types of wastewater, maintaining consistent sensitivity (0.64 ± 0.10 mA L mg−1 m−2 with synthetic wastewater and 0.78 ± 0.13 mA L mg−1 m−2 with urban wastewater) and limit of detection (49 ± 8 mg L−1 for synthetic wastewater and 44 ± 7 mg L−1 for urban wastewater). Crucially, pre-sterilization, conductivity adjustments, and nitrogen purging of wastewater are not required before its introduction into the biosensor. However, the presence of native aerobic microorganisms in the wastewater might affect the current output. Metagenomics and taxonomic analyses revealed that the alterations in biofilm composition are predominantly in response to the varied chemical and microbiological compositions of different substrates. Despite variations in anodic biofilm composition, the MFC-based biosensor maintains a relative error comparable to the standard BOD test. This highlights the resilience and flexibility of the biosensor when directly used with a variety of wastewater types before full biofilm adjustment.

Towards rapid formation of electroactive biofilm: insights from thermodynamics and electric field manipulation

Electroactive biofilm (EAB) has garnered significant attention due to its effectiveness in pollutant remediation, electricity generation, and chemical synthesis. However, achieving precise control over the rapid formation of EAB presents challenges for the practical implementation of bioelectrochemical technology. In this study, we investigated the regulation of EAB formation by manipulating applied electric potential. We developed a modified XDLVO model for the applied electric field and quantitatively assessed the feasibility of existing rapid formation strategies for EAB. Our results revealed that electrostatic (EL) force significantly influenced EAB formation in the presence of the applied electric field, with the potential difference between the electrode and the microbial solution being the primary determinant of EL force. Compared to -0.2 V and 0 V vs.Ag/AgCl, EAB exhibited the highest electrochemical performance at 0.2 V vs.Ag/AgCl, with a maximum current density of 6.044 ± 0.10 A/m2, surpassing that at -0.2 V vs.Ag/AgCl and 0 V vs.Ag/AgCl by 1.73 times and 1.31 times, respectively. Furthermore, EAB demonstrated the highest biomass accumulation, measuring a thickness of 25 ± 2 μm at 0.2 V vs. Ag/AgCl, representing increases of 1.67 and 1.25 times compared to -0.2 V vs.Ag/AgCl and 0 V vs.Ag/AgCl, respectively. The strong electrostatic attraction under the anodic potential promoted the formation of a monolayer of biofilm. Additionally, the hydrophilicity and hydrophobicity of the biofilm were altered following inversion culture. The Lewis acid-base (AB) attraction offset the electrostatic repulsion caused by negative charges, it is beneficial for the formation of biofilms. This study, for the first time, elucidated the difference in the formation of cathode and anode biofilm from a thermodynamic perspective in the context of electric field introduction, laying the theoretical foundation for the directional regulation of the rapid formation of typical electroactive biofilms.

Inoculum source determines the stress resistance of electroactive functional taxa in biofilms: A metagenomic perspective

The inoculum has a crucial impact on bioreactor initialization and performance. However, there is currently a lack of guidance on selecting appropriate inocula for applications in environmental biotechnology. In this study, we applied microbial electrolysis cells (MECs) as models to investigate the differences in the functional potential of electroactive microorganisms (EAMs) within anodic biofilms developed from four different inocula (natural or artificial), using shotgun metagenomic techniques. We specifically focused on extracellular electron transfer (EET) function and stress resistance, which affect the performance and stability of MECs. Community profiling revealed that the family Geobacteraceae was the key EAM taxon in all biofilms, with Geobacter as the dominant genus. The c-type cytochrome gene imcH showed universal importance for Geobacteraceae EET and was utilized as a marker gene to evaluate the EET potential of EAMs. Additionally, stress response functional genes were used to assess the stress resistance potential of Geobacter species. Comparative analysis of imcH gene abundance revealed that EAMs with comparable overall EET potential could be enriched from artificial and natural inocula (P > 0.05). However, quantification of stress response gene copy numbers in the genomes demonstrated that EAMs originating from natural inocula possessed superior stress resistance potential (196 vs. 163). Overall, this study provides novel perspectives on the inoculum effect in bioreactors and offers theoretical guidance for selecting inoculum in environmental engineering applications.

Unravelling the mechanisms of organic micropollutant removal in bio-electrochemical systems: Insights into sorption, electrochemical degradation, and biodegradation processes

Bio-electrochemical systems (BESs) have recently been proposed as an efficient treatment technology to remove organic micropollutants from water treatment plants. In this study, we aimed to differentiate between sorption, electrochemical transport/degradation, and biodegradation. Using electro-active microorganisms and electrodes, we investigated organic micropollutant removal at environmentally relevant concentrations, clarifying the roles of sorption and electrochemical and biological degradation. The role of anodic biofilms on the removal of 10 relevant organic micropollutants was studied by performing separate sorption experiments on carbon-based electrodes (graphite felt, graphite rod, graphite granules, and granular activated carbon) and electrochemical degradation experiments at two different electrode potentials (−0.3 and 0 V).Granular activated carbon showed the highest sorption of micropollutants; applying a potential to graphite felt electrodes increased organic micropollutant removal. Removal efficiencies >80 % were obtained for all micropollutants at high anode potentials (+0.955 V), indicating that the studied compounds were more susceptible to oxidation than to reduction. All organic micropollutants showed removal when under bio-electrochemical conditions, ranging from low (e.g. metformin, 9.3 %) to exceptionally high removal efficiencies (e.g. sulfamethoxazole, 99.5 %). The lower removal observed under bio-electrochemical conditions when compared to only electrochemical conditions indicated that sorption to the electrode is key to guarantee high electrochemical degradation. The detection of transformation products of chloridazon and metformin indicated that (bio)-electrochemical degradation occurred. This study confirms that BES can treat some organic micropollutants through several mechanisms, which merits further investigation.

Modulating anodic performance of halophilic electroactive microbial consortia with selective enrichment using metal-based terminal electron acceptors

Long start-up times and lack of robust halophilic electroactive microbes (EAM) challenge the implementation of microbial fuel cells (MFC) for treating saline wastewater. Here a pragmatic strategy is presented to selectively enrich halophilic EAM consortium capitalising on their cellular respiration process using graphene oxide (GO) and metal ions (Fe3+, Mn4+ and Cu2+) as the primary and secondary terminal electron acceptors (TEA), respectively. Enrichment of GO-respiring EAM consortium with Fe3+ ions enabled growth of an efficient anodic biofilm within a short start-up time of 50 h and generated up to 1.1 W/m3 while treating highly saline electrolyte (2 % NaCl). In conjunction with predictive genetic profiling, in-depth electrochemical analysis of the biofilm unravelled key mechanisms of direct extracellular electron transfer for generating high anodic currents. Additionally, the inclusion of Fe3+ ions as secondary TEA upregulated the metabolic activity of GO-respiring EAM, which subsequently influenced their anodic performance rather than the relative abundance of exoelectrogens in the biofilm. Thus, this study stimulates the discovery of novel EAM communities using an advanced and sensible design of selective media by varying the electron donor and TEA couplings.

Investigating the bacterial consortia properties of electrogenic anodic biofilm in a double-chamber microbial fuel cell: electrochemical, physical, biochemical and molecular characterization

This work evaluated the electrochemical, physical, biochemical, and molecular characterization of electrogens from a graphite felt anode when zinc oxide on activated carbon (ZnO/AC) was used as a cathodic electrocatalyst in a double-chambered microbial fuel cell (DCMFC). The electrochemical polarization behavior of the DCMFC showed that ZnO/AC had a higher power density (PDmax) of 89 mW m−2 with a corresponding cell current density (CD) of 248 mA m−2 and a voltage output of 395 mV, which was higher than those of the blank electrode used as a benchmark (PDmax of 68 mW m−2 at a CD of 161 mA m−2 and a voltage of 421 mV). Furthermore, scanning electron microscopy and transmission electron microscopy revealed that the morphology and interior properties of the strains varied among the rods (bacilli), spirals (vibrios), and spheres (diplococci, staphylococci and streptococci). In addition, biochemical characterization via the Vitek2 compact system and molecular analysis via 16 S rRNA and 18 S rRNA gene sequencing revealed the occurrence of nine prevalent species that were correlated with Sphingobacterium spiritivorum, Ochrobactrum anthropicus, Pseudomonas mendocina, Stenotrophomonas maltophilia, Leuconostoc mesenteroides, Staphylococcus equorum, Bacillus subtilis HQ334981.1, Kocuria kristinae KC581674.1 and Saccharomyces cerevisiae NR111007.1. Consequently, the present study outlines different characterization strategies for electrogenic microbes that play an important role in the overall performance of DCMFC for scaling up and managing existing environmental pollution for sustainable energy generation.

Efficient Enhancement of Extracellular Electron Transfer in Shewanella oneidensis MR-1 via CRISPR-Mediated Transposase Technology.

Electroactive bacteria, exemplified by Shewanella oneidensis MR-1, have garnered significant attention due to their unique extracellular electron-transfer (EET) capabilities, which are crucial for energy recovery and pollutant conversion. However, the practical application of MR-1 is constrained by its EET efficiency, a key limiting factor, due to the complexity of research methodologies and the challenges associated with the practical use of gene editing tools. To address this challenge, a novel gene integration system, INTEGRATE, was developed, utilizing CRISPR-mediated transposase technologies for precise genomic insertion within the S. oneidensis MR-1 genome. This system facilitated the insertion of extensive gene segments at different sites of the Shewanella genome with an efficiency approaching 100%. The inserted cargo genes could be kept stable on the genome after continuous cultivation. The enhancement of the organism's EET efficiency was realized through two primary strategies: the integration of the phenazine-1-carboxylic acid synthesis gene cluster to augment EET efficiency and the targeted disruption of the SO3350 gene to promote anodic biofilm development. Collectively, our findings highlight the potential of utilizing the INTEGRATE system for strategic genomic alterations, presenting a synergistic approach to augment the functionality of electroactive bacteria within bioelectrochemical systems.

A combination of microbial electrolysis cells and bioaugmentation can effectively treat synthetic wastewater containing polycyclic aromatic hydrocarbon.

The anaerobic biodegradation of polycyclic aromatic hydrocarbons (PAHs) is challenging due to its toxic effect on the microbes. Microbial electrolysis cells (MECs), with their excellent characteristics of anodic and cathodic biofilms, can be a viable way to enhance the biodegradation of PAHs. This work assessed different cathode materials (carbon brush and nickel foam) combined with bioaugmentation on typical PAHs-naphthalene biodegradation and analyzed the inhibition amendment mechanism of microbial biofilms in MECs. Compared with the control, the degradation efficiency of naphthalene with the nickel foam cathode supplied with bioaugmentation dosage realized a maximum removal rate of 94.5 ± 3.2%. The highest daily recovered methane yield (227 ± 2 mL/gCOD) was also found in the nickel foam cathode supplied with bioaugmentation. Moreover, the microbial analysis demonstrated the significant switch of predominant PAH-degrading microorganisms from Pseudomonas in control to norank_f_Prolixibacteraceae in MECs. Furthermore, hydrogentrophic methanogenesis prevailed in MEC reactors, which is responsible for methane production. This study proved that MEC combined with bioaugmentation could effectively alleviate the inhibition of PAH, with the nickel foam cathode obtaining the fastest recovery rate in terms of methane yield.

Asymmetric reactors as an innovative approach for optimum microbial fuel cells performance

Microbial fuel cells (MFCs) exhibit asymmetric overpotentials and redox potentials/rates at the cathode and anode. However, the impact of this asymmetric on power density and anode microbial community remains poorly understood. In this study, asymmetric reactors were designed for H-type MFCs to investigate this configuration. Contrary to the initial expectation of increased MFC power output with larger cathode volumes, it was unexpectedly found that the small-cathode reactor outperformed the large-cathode reactor, achieving a 61 % increase in maximum power density. Electrochemical characterization revealed a slightly lower charge transfer resistance (29.34 Ω) in the small-cathode reactor with carbon-felt anode biofilm in PBS. Moreover, 16S rRNA sequence analysis showed that the small-cathode reactor harbored a higher proportion of anaerobic bacteria (83 %), lower species diversity, and a higher abundance of exoelectrogens. Additionally, higher abundances of key gene modules (top eight), such as quinone oxidoreductase and citrate cycle, were observed in the small-cathode reactor. The anode biofilms in both reactors also synthesized some vitamins, such as menaquinone and thiamine. Furthermore, compared to the large-cathode reactor, each set of the small-cathode reactor saved ¥ 20, 23 g of borosilicate, and 55 mL of cathode electrolytes. This study sheds light on the interplay between reactor conformation and performance, contributing to the development of low-cost, high-performance MFCs for real field conditions.

Bioelectrochemical anaerobic digestion mitigates microplastic pollution and promotes methane recovery of wastewater treatment in biofilm system

Bioelectrochemical systems (BES) offer significant potential for treating refractory waste and recovering bioenergy. However, their ability to mitigate microplastic pollution in wastewater remains unexplored. This study showed that BES facilitated the treatment of polyethylene (PE), polyvinyl chloride (PVC), and Mix (PE+PVC) microplastic wastewater and the methane recovery (40.61%, 20.02%, 21.19%, respectively). The lactate dehydrogenase (LDH), adenosine triphosphate (ATP), cytochrome c, and nicotinamide adenine dinucleotide (NADH/NAD+) ratios were elevated with electrical stimulation. Moreover, the applied voltage improved the polysaccharides content of the extracellular polymeric substances (EPS) in the PE-BES but decreased in PVC-BES, while the proteins showed the opposite trend. Metatranscriptomic sequencing showed that the abundance of fermentation bacteria, acetogens, electrogens, and methanogens was greatly enhanced by applying voltage, especially at the anode. Methane metabolism was dominated by the acetoclastic methanogenic pathway, with the applied voltage promoting the enrichment of Methanothrix, resulting in the direct conversion of acetate to acetyl-CoA via acetate-CoA ligase (EC: 6.2.1.1), and increased metabolic activity in the anode. Moreover, applied voltage greatly boosted the function genes expression level related to energy metabolism, tricarboxylic acid (TCA) cycle, electron transport, and transporters on the anode biofilm. Overall, these results demonstrate that BES can mitigate microplastic pollution during wastewater treatment.

Sediment microbial fuel cell with biochar-modified cathode for remediation of black-odorous water sediments and analysis of microbial community

Sediment microbial fuel cells (SMFCs) generate electricity through microbial degradation of organic matter in sediments, and the cathode catalyst used for the oxygen reduction reaction is a key factor affecting the performance of SMFCs. To improve power production in SMFCs and remediate black-odorous water sediments, this study modifies the cathode carbon felt of the SMFC system with Enteromorpha prolifera (EP) biochar with high N and P content. The study demonstrated that the SMFC system with the modified cathode carbon felt (C-800℃/CF) exhibited superior power generation capabilities, achieving a maximum power density of 219.44 mW·m−2. Over 25 days of operation, the anode biofilm effectively degraded around 78.15% of total organic carbon, 79.55% of acid volatile sulfide, 74.78% of total nitrogen, 80.58% of ammonia nitrogen, and 77.46% of total phosphorus in the sediment. In addition, microbial community analysis showed that functional bacteria Thiobacillus, Acidaminobacter, Sulfurimonas, Bacteroidetes_vadinHA17, and Sideroxydans were the dominant genera in the anode biofilm.

Unveiling microbial degradation of triclosan: Degradation mechanism, pathways, and catalyzing clean energy

Emerging organic contaminants present in the environment can be biodegraded in anodic biofilms of microbial fuel cells (MFCs). However, there is a notable gap existing in deducing the degradation mechanism, intermediate products, and the microbial communities involved in degradation of broad-spectrum antibiotic such as triclosan (TCS). Herein, the possible degradation of TCS is explored using TCS acclimatized biofilms in MFCs. 95% of 5 mgL−1 TCS are been biodegraded within 84 h with a chemical oxygen demand (COD) reduction of 62% in an acclimatized-MFC (A-MFC). The degradation of TCS resulted in 8 intermediate products including 2,4 -dichlorophenol which gets further mineralized within the system. Concurrently, the 16S rRNA V3–V4 sequencing revealed that there is a large shift in microbial communities after TCS acclimatization and MFC operation. Moreover, 30 dominant bacterial species (relative intensity >1%) are identified in the biofilm in which Sulfuricurvum kujiense, Halomonas phosphatis, Proteiniphilum acetatigens, and Azoarcus indigens significantly contribute to dihydroxylation, ring cleavage and dechlorination of TCS. Additionally, the MFC was able to produce 818 ± 20 mV voltage output with a maximum power density of 766.44 mWm−2. The antibacterial activity tests revealed that the biotoxicity of TCS drastically reduced in the MFC effluent, signifying the non-toxic nature of the degraded products. Hence, this work provides a proof−of−concept strategy for sustainable mitigation of TCS in wastewaters with enhanced bioelectricity generation.

Anaerobic treatment of groundwater co-contaminated by toluene and copper in a single chamber bioelectrochemical system

Addressing the simultaneous removal of multiple coexisting groundwater contaminants poses a significant challenge, primarily because of their different physicochemical properties. Indeed, different chemical compounds may necessitate establishing distinct, and sometimes conflicting, (bio)degradation and/or removal pathways. In this work, we investigated the concomitant anaerobic treatment of toluene and copper in a single-chamber bioelectrochemical cell with a potential difference of 1 V applied between the anode and the cathode. As a result, the electric current generated by the bioelectrocatalytic oxidation of toluene at the anode caused the abiotic reduction and precipitation of copper at the cathode, until the complete removal of both contaminants was achieved. Open circuit potential (OCP) experiments confirmed that the removal of copper and toluene was primarily associated with polarization. Analogously, abiotic experiments, at an applied potential of 1 V, confirmed that neither toluene was oxidized nor copper was reduced in the absence of microbial activity. At the end of each experiment, both electrodes were characterized by means of a comprehensive suite of chemical and microbiological analyses, evidencing a highly selected microbial community competent in the biodegradation of toluene in the anodic biofilm, and a uniform electrodeposition of spherical Cu2O nanoparticles over the cathode surface.

Impact of bioelectricity on DNRA process and microbial community composition within cathodic biofilms in dual-chambered bioelectrode microbial fuel cell (MFC)

The synchronous bioelectricity generation and dissimilatory nitrate reduction to ammonium (DNRA) pathway in Klebsiella variicola C1 was investigated. The presence of bioelectricity facilitated cell growth on the anodic biofilms, consequently enhancing the nitrate removal efficiency decreasing total nitrogen levels and causing a negligible accumulation of NO2– in the supernatant. Genomic analysis revealed that K. variicola C1 possessed a complete DNRA pathway and largely annotated electron shuttles. The up-regulated expression of genes narG and nirB, encoding nitrite oxidoreductase and nitrite reductase respectively, was closely associated with increased extracellular electron transfer (EET). High-throughput sequencing analysis was employed to investigate the impact of bioelectricity on microbial community composition within cathodic biofilms. Results indicated that Halomonas, Marinobacter and Prolixibacteraceae were enriched at the cathode electrodes. In conclusion, the integration of a DNRA strain with MFC facilitated the efficient removal of wastewater containing high concentrations of NO3– and enabled the environmentally friendly recovery of NH4+.

Significance of a minor pilin PilV in biofilm cohesion of Geobacter sulfurreducens

Bacterial adhesion plays a vital role in forming and shaping the structure of electroactive biofilms that are essential for the performance of bioelectrochemical systems (BESs). Type IV pili are known to mediate cell adhesion in many Gram-negative bacteria, but the mechanism of pili-mediated cell adhesion of Geobacter species on anode surface remains unclear. Herein, a minor pilin PilV2 was found to be essential for cell adhesion ability of Geobacter sulfurreducens since the lack of pilV2 gene depressed the cell adhesion capability by 81.2% in microplate and the anodic biofilm density by 23.1 % at −0.1 V and 37.7 % at −0.3 V in BESs. The less cohesiveness of mutant biofilms increased the charge transfer resistance and biofilm resistance, which correspondingly lowered current generation of the pilV2-deficient strain by up to 63.2 % compared with that of the wild-type strain in BESs. The deletion of pilV2 posed an insignificant effect on the production of extracellular polysaccharides, pili, extracellular cytochromes and electron shuttles that are involved in biofilm formation or extracellular electron transfer (EET) process. This study demonstrated the significance of pilV2 gene in cell adhesion and biofilm formation of G. sulfurreducens, as well as the importance of pili-mediated adhesion for EET of electroactive biofilm.

Low temperature acclimation of electroactive microorganisms may be an effective strategy to enhance the toxicity sensing performance of microbial fuel cell sensors

Microbial fuel cell (MFC) sensing is a promising method for real-time detection of water biotoxicity, however, the low sensing sensitivity limits its application. This study adopted low temperature acclimation as a strategy to enhance the toxicity sensing performance of MFC biosensor. Two types of MFC biosensors were started up at low (10 °C) or warm (25 °C) temperature, denoted as MFC-Ls and MFC-Ws respectively, using Pb2+ as the toxic substance. MFC-Ls exhibited superior sensing sensitivities towards Pb2+ compared with MFC-Ws at both low (10 °C) and warm (25 °C) operation temperatures. For example, the inhibition rate of voltage of MFC-Ls was 22.81 % with 1 mg/L Pb2+ shock at 10 °C, while that of MFC-Ws was only 5.9 %. The morphological observation showed the anode biofilm of MFC-Ls had appropriate amount of extracellular polymer substances, thinner thickness (28.95 μm for MFC-Ls and 41.58 μm for MFC-Ws) and higher proportion of living cells (90.65 % for MFC-Ls and 86.01 % for MFC-Ws) compared to that of MFC-Ws. Microbial analysis indicated the enrichment of psychrophilic electroactive microorganisms and cold-active enzymes as well as their sensitivity to Pb2+ shock was the foundation for the effective operation and good performance of MFC-Ls biosensors. In conclusion, low temperature acclimation of electroactive microorganisms enhanced not only the sensitivity but also the temperature adaptability of MFC biosensors.

Elucidating the development of cooperative anode-biofilm-structures

Microbial electrochemical systems are a highly versatile platform technology with a particular focus on the interplay of chemical and electrical energy conversion and offer immense potential for a sustainable bioeconomy. The industrial realization of this potential requires a critical focus on biofilm optimization if performance is to be controlled over a long period of time. Moreover, the aspect and influence of cooperativity has to be addressed as many applied anodic bioelectrochemical systems will most likely be operated with a diversity of interacting microbial species. Hence, the aim of this study was to analyze how interspecies dependence and cooperativity of a model community influence the development of anodic biofilms. To investigate biofilm activity in a spatially resolved manner, a microfluidic bioelectrochemical flow cell was developed that can be equipped with user-defined electrode materials and operates under laminar flow conditions. With this infrastructure, the development of single and co-culture biofilms of the two model organisms Shewanella oneidensis and Geobacter sulfurreducens on graphite electrodes was monitored by optical coherence tomography analysis. The interdependence in the co-culture biofilm was achieved by feeding the community with lactate, which is converted by S. oneidensis into acetate, which in turn serves as substrate for G. sulfurreducens. The results show that co-cultivation resulted in the formation of denser biofilms than in single culture. Moreover, we hypothesize that S. oneidensis in return utilizes the conductive biofilm matrix build by G. sulfurreducens for direct interspecies electron transfer (DIET) to the anode. FISH analysis revealed that the biofilms consisted of approximately two-thirds G. sulfurreducens cells, which most likely formed a conductive 3D network throughout the biofilm matrix, in which evenly distributed tubular S. oneidensis colonies were embedded without direct contact to the anode surface. Live/dead staining shows that the outermost biofilm contained almost exclusively dead cells (98 %), layers near the anode contained 45–56 % and the entire biofilm contained 82 % live cells. Our results exemplify how the architecture of the exoelectrogenic biofilm dynamically adapts to the respective process conditions.

PDA-Fe3O4 decorated carbon felt anode enhancing electrochemical performance of microbial fuel cells: Effect of electrode materials on electroactive biofilm

Anode modification is an effective strategy for enhancing the electrochemical performance of microbial fuel cell (MFC). However, the impacts of the modified materials on anode biofilm development during MFC operation have been less studied. We prepared a novel PDA-Fe3O4-CF composite anode by coating original carbon felt anode (CF) with polydopamine (PDA) and Fe3O4 nanoparticles. The composite anode material was characterized by excellent hydrophilicity and electrical conductivity, and the anodic biofilm exhibited fast start-up, higher biomass, and more uniform biofilm layer after MFC operation. The MFC reactor assembled with the composite anode achieved a maximum power density of 608 mW m−2 and an output voltage of 586 mV, which were 316.4% and 72.4% higher than the MFC with the original CF anode, respectively. Microbial community analysis indicated that the modified anode biofilm had a higher relative abundance of exoelectrogen species in comparison to the unmodified anode. The PICRUSt data revealed that the anodic materials may affect the bioelectrochemical performance of the biofilm by influencing the expression levels of key enzyme genes involved in biofilm extracellular polymer (EPS) secretion and extracellular electron transfer (EET). The growth of the anodic biofilm would exert positive or negative influences on the efficiency of electricity production and electron transfer of the MFCs at different operating stages. This work expands the knowledge of the role that anodic materials play in the development and electrochemical performance of anodic biofilm in MFCs.