Characterization of melanin from Exophiala mesophila with the prospect of potential biotechnological applications.

Introducion: Fungal melanin is an underexplored natural biomaterial of great biotechnological interest in different areas. This study investigated the physical, chemical, electrochemical, and metal-binding properties of melanin extracted from the metallotolerant black fungus Exophiala mesophila strain IRTA-M2-F10. Materials and methods: Specific inhibitory studies with tricyclazole and biochemical profiling of whole cells by synchrotron radiation-based Fourier-transform infrared spectral microscopy (SR-FTIRM) were performed. An optimized extraction protocol was implemented, and purified fungal melanin was characterized using an array of spectrophotometric techniques (UV-Vis, FTIR, and EPR) and by cyclic voltammetry (CV) experiments. The metal-binding capacity of melanin extracts was also assessed by using Cr(VI) as a model heavy metal. Results: Inhibitory studies indicated that 1,8-dihydroxynaphthalene may be the main precursor molecule of E. mesophila melanin (DHN-melanin). The biochemical characterization of fungal melanin extracts were benchmarked against those from two melanins comprising the precursor molecule L-3,4-dihydroxiphenylalanine (DOPA-melanin): extracts from the ink of the cephalopod Sepia officinalis and DOPA-melanin synthesized in the laboratory. The CV results of melanin extracts incubated with and without cell suspensions of the electroconductive bacterium Geobacter sulfurreducens were indicative of novel semiquinone/hydroquinone redox transformations specific for each melanin type. These interactions may play an important role in cation exchange for the adsorption of metals and in microbial interspecies electron transfer processes. Discussion: The obtained results provided further evidence for the DHN-nature of E. mesophila melanin. The FTIR profiling of melanin extracts exposed to Cr(VI), compared to unexposed melanin, resulted in useful information on the distinct surface-binding properties of fungal melanin. The parameters of the Langmuir and Freundlicht isotherms for the adsorption of Cr(VI) were determined and compared to bibliographic data. Altogether, the inherent properties of fungal melanin suggest its promising potential as a biomaterial for environmental applications.

Novel electrochemical strategies for the microbial conversion of CO2 into biomass and volatile fatty acids using a fluid-like bed electrode in a three-phase reactor.

Microbial electrosynthesis (MES) constitutes a bioelectrochemical process where bacteria uptake electrons extracellularly from a polarized electrode to incorporate them into their anabolic metabolism. However, the efficiency of current MES reactor designs can be lower than expected due to limitations regarding electron transfer and mass transport. One of the most promising bioreactor configurations to overcome these bottlenecks is the Microbial Electrochemical Fluidized Bed Reactor (ME-FBR). In this study, microbial CO2 fixation is investigated for the first time in a ME-FBR operated as a 3-phase reactor (solid-liquid-gas). An electroconductive carbon bed, acting as a working electrode, was fluidized with gas and polarized at different potentials (-0.6, -0.8 and -1 V vs. Ag/AgCl) so it could act as an electron donor (biocathode). Under these potentials, CO2 fixation and electron transfer were evaluated. Autotrophic electroactive microorganisms from anaerobic wastewater were enriched in a ME-FBR in the presence of 2-bromoethanosulfonic acid (BES) to inhibit the growth of methanogens. Cyclic voltammetry analysis revealed interaction between the microorganisms and the cathode. Furthermore, volatile fatty acids like propionate, formate and acetate were detected in the culture supernatant. Acetate production had a maximum rate of ca. 1 g L-1 day-1 . Planktonic cell biomass was produced under continuous culture at values as high as ca. 0.7 g L-1 dry weight. Overall, this study demonstrates the feasibility of employing a fluidized electrode with gaseous substrates and electricity as the energy source for generating biomass and carboxylic acids.

Water treatment and reclamation by implementing electrochemical systems with constructed wetlands.

Seasonal or permanent water scarcity in off-grid communities can be alleviated by recycling water in decentralized wastewater treatment systems. Nature-based solutions, such as constructed wetlands (CWs), have become popular solutions for sanitation in remote locations. Although typical CWs can efficiently remove solids and organics to meet water reuse standards, polishing remains necessary for other parameters, such as pathogens, nutrients, and recalcitrant pollutants. Different CW designs and CWs coupled with electrochemical technologies have been proposed to improve treatment efficiency. Electrochemical systems (ECs) have been either implemented within the CW bed (ECin-CW) or as a stage in a sequential treatment (CW + EC). A large body of literature has focused on ECin-CW, and multiple scaled-up systems have recently been successfully implemented, primarily to remove recalcitrant organics. Conversely, only a few reports have explored the opportunity to polish CW effluents in a downstream electrochemical module for the electro-oxidation of micropollutants or electro-disinfection of pathogens to meet more stringent water reuse standards. This paper aims to critically review the opportunities, challenges, and future research directions of the different couplings of CW with EC as a decentralized technology for water treatment and recovery.

Fluid-like cathode enhances valuable biomass production from brewery wastewater in purple phototrophic bacteria.

The climate crisis requires rethinking wastewater treatment to recover resources, such as nutrients and energy. In this scenario, purple phototrophic bacteria (PPB), the most versatile microorganisms on earth, are a promising alternative to transform the wastewater treatment plant concept into a biorefinery model by producing valuable protein-enriched biomass. PPB are capable of interacting with electrodes, exchanging electrons with electrically conductive materials. In this work, we have explored for mobile-bed (either stirred or fluidized) cathodes to maximize biomass production. For this purpose, stirred-electrode reactors were operated with low-reduced (3.5 e-/C) and high-reduced (5.9 e-/C) wastewater under cathodic polarization (-0.4 V and -0.8 V vs. Ag/AgCl). We observed that cathodic polarization and IR irradiation can play a key role in microbial and phenotypic selection, promoting (at -0.4 V) or minimizing (at -0.8 V) the presence of PPB. Then, we further study how cathodic polarization modulates PPB biomass production providing a fluid-like electrode as part of a so-called photo microbial electrochemical fluidized-bed reactor (photoME-FBR). Our results revealed the impact of reduction status of carbon source in wastewater to select the PPB photoheterotrophic community and how electrodes drive microbial population shifts depending on the reduction status of such carbon source.

Electrochemically assisted production of biogenic palladium nanoparticles for the catalytic removal of micropollutants in wastewater treatment plants effluent.

Biogenic palladium nanoparticles (bio-Pd NPs) are used for the reductive transformation and/or dehalogenation of persistent micropollutants. In this work, H2 (electron donor) was produced in situ by an electrochemical cell, permitting steered production of differently sized bio-Pd NPs. The catalytic activity was first assessed by the degradation of methyl orange. The NPs showing the highest catalytic activity were selected for the removal of micropollutants from secondary treated municipal wastewater. The synthesis at different H2 flow rates (0.310 L/hr or 0.646 L/hr) influenced the bio-Pd NPs size. The NPs produced over 6 hr at a low H2 flow rate had a larger size (D50 = 39.0 nm) than those produced in 3 hr at a high H2 flow rate (D50 = 23.2 nm). Removal of 92.1% and 44.3% of methyl orange was obtained after 30 min for the NPs with sizes of 39.0 nm and 23.2 nm, respectively. Bio-Pd NPs of 39.0 nm were used to treat micropollutants present in secondary treated municipal wastewater at concentrations ranging from µg/L to ng/L. Effective removal of 8 compounds was observed: ibuprofen (69.5%) < sulfamethoxazole (80.6%) < naproxen (81.4%) < furosemide (89.7%) < citalopram (91.7%) < diclofenac (91.9%) < atorvastatin (> 94.3%) < lorazepam (97.2%). Removal of fluorinated antibiotics occurred at > 90% efficiency. Overall, these data indicate that the size, and thus the catalytic activity of the NPs can be steered and that the removal of challenging micropollutants at environmentally relevant concentrations can be achieved through the use of bio-Pd NPs.

Microbial photoelectrosynthesis: Feeding purple phototrophic bacteria electricity to produce bacterial biomass.

Purple phototrophic bacteria are one of the main actors in chemolithotrophic carbon fixation and, therefore, fundamental in the biogeochemical cycle. These microbes are capable of using insoluble electron donors such as ferrous minerals or even carbon-based electrodes. Carbon fixation through extracellular electron uptake places purple phototrophic bacteria in the field of microbial electrosynthesis as key carbon capturing microorganisms. In this work we demonstrate biomass production dominated by purple phototrophic bacteria with a cathode (-0.6 V vs. Ag/AgCl) as electron donor. In addition, we compared the growth and microbial population structure with ferrous iron as the electron donor. We detect interaction between the cathode and the consortium showing a midpoint potential of 0.05 V (vs. Ag/AgCl). Microbial community analyses revealed different microbial communities depending on the electron donor, indicating different metabolic interactions. Electrochemical measurements together with population analyses point to Rhodopseudomonas genus as the key genus in the extracellular electron uptake. Furthermore, the genera Azospira and Azospirillum could play a role in the photoelectrotrophic consortium.

A review on antibiotics removal: Leveraging the combination of grey and green techniques.

Antibiotics are currently a major source of concern around the world due to the serious risks posed to human health and the environment. The performance of the secondary wastewater treatment processes/technologies (representing grey process) and constructed wetlands (CWs) (typical green process) in removing antibiotics and antibiotic resistance genes (ARG) was reviewed. The result showed that the grey process mainly removes antibiotics, but does not significantly remove ARG, and some processes may even cause ARG enrichment. The overall treatment in CWs is better than WWTPs, especially for ARG. Vertical subsurface flow CWs (VFCWs) are more conductive to antibiotics removal, while horizontal subsurface flow CWs (HFCWs) have a better ARG removal. More importantly, this review admits and suggests that the combination of grey process with green process is an effective strategy to remove antibiotics and ARG. The most advantage of the combination lies in realizing complementary advantages, i.e. the grey process as the primary treatment while CWs as the polishing stage. The efficiency of such the hybrid system is much higher than either single treatment process.

Microbial Electrochemically Assisted Treatment Wetlands: Current Flow Density as a Performance Indicator in Real-Scale Systems in Mediterranean and Northern European Locations.

A METland is an innovative treatment wetland (TW) that relies on the stimulation of electroactive bacteria (EAB) to enhance the degradation of pollutants. The METland is designed in a short-circuit mode (in the absence of an external circuit) using an electroconductive bed capable of accepting electrons from the microbial metabolism of pollutants. Although METlands are proven to be highly efficient in removing organic pollutants, the study of in situ EAB activity in full-scale systems is a challenge due to the absence of a two-electrode configuration. For the first time, four independent full-scale METland systems were tested for the removal of organic pollutants and nutrients, establishing a correlation with the electroactive response generated by the presence of EAB. The removal efficiency of the systems was enhanced by plants and mixed oxic-anoxic conditions, with an average removal of 56 g of chemical oxygen demand (COD) mbed material -3 day-1 and 2 g of total nitrogen (TN) mbed material -3 day-1 for Ørby 2 (partially saturated system). The estimated electron current density (J) provides evidence of the presence of EAB and its relationship with the removal of organic matter. The tested METland systems reached the max. values of 188.14 mA m-2 (planted system; IMDEA 1), 223.84 mA m-2 (non-planted system; IMDEA 2), 125.96 mA m-2 (full saturated system; Ørby 1), and 123.01 mA m-2 (partially saturated system; Ørby 2). These electron flow values were remarkable for systems that were not designed for energy harvesting and unequivocally show how electrons circulate even in the absence of a two-electrode system. The relation between organic load rate (OLR) at the inlet and coulombic efficiency (CE; %) showed a decreasing trend, with values ranging from 8.8 to 53% (OLR from 2.0 to 16.4 g COD m-2 day-1) for IMDEA systems and from 0.8 to 2.5% (OLR from 41.9 to 45.6 g COD m-2 day-1) for Ørby systems. This pattern denotes that the treatment of complex mixtures such as real wastewater with high and variable OLR should not necessarily result in high CE values. METland technology was validated as an innovative and efficient solution for treating wastewater for decentralized locations.

Microbial Electrochemical Fluidized Bed Reactor: A Promising Solution for Removing Pollutants From Pharmaceutical Industrial Wastewater.

The capacity of electroactive bacteria to exchange electrons with electroconductive materials has been explored during the last two decades as part of a new field called electromicrobiology. Such microbial metabolism has been validated to enhance the bioremediation of wastewater pollutants. In contrast with standard materials like rods, plates, or felts made of graphite, we have explored the use of an alternative strategy using a fluid-like electrode as part of a microbial electrochemical fluidized bed reactor (ME-FBR). After verifying the low adsorption capacity of the pharmaceutical pollutants on the fluid-bed electrode [7.92 ± 0.05% carbamazepine (CBZ) and 9.42 ± 0.09% sulfamethoxazole (SMX)], our system showed a remarkable capacity to outperform classical solutions for removing pollutants (more than 80%) from the pharmaceutical industry like CBZ and SMX. Moreover, the ME-FBR performance revealed the impact of selecting an anode potential by efficiently removing both pollutants at + 200 mV. The high TOC removal efficiency also demonstrated that electrostimulation of electroactive bacteria in ME-FBR could overcome the expected microbial inhibition due to the presence of CBZ and SMX. Cyclic voltammograms revealed the successful electron transfer between microbial biofilm and the fluid-like electrode bed throughout the polarization tests. Finally, Vibrio fischeri-based ecotoxicity showed a 70% reduction after treating wastewater with a fluid-like anode (+ 400 mV), revealing the promising performance of this bioelectrochemical approach.

Assessing METland® Design and Performance Through LCA: Techno-Environmental Study With Multifunctional Unit Perspective.

Conventional wastewater treatment technologies are costly and energy demanding; such issues are especially remarkable when small communities have to clean up their pollutants. In response to these requirements, a new variety of nature-based solution, so-called METland®, has been recently develop by using concepts from Microbial Electrochemical Technologies (MET) to outperform classical constructed wetland regarding wastewater treatment. Thus, the current study evaluates two operation modes (aerobic and aerobic-anoxic) of a full-scale METland®, including a Life Cycle Assessment (LCA) conducted under a Net Environmental Balance perspective. Moreover, a combined technical and environmental analysis using a Net Eutrophication Balance (NEuB) focus concluded that the downflow (aerobic) mode achieved the highest removal rates for both organic pollutant and nitrogen, and it was revealed as the most environmentally friendly design. Actually, aerobic configuration outperformed anaero/aero-mixed mode in a fold-range from 9 to 30%. LCA was indeed recalculated under diverse Functional Units (FU) to determine the influence of each FU in the impacts. Furthermore, in comparison with constructed wetland, METland® showed a remarkable increase in wastewater treatment capacity per surface area (0.6 m2/pe) without using external energy. Specifically, these results suggest that aerobic-anoxic configuration could be more environmentally friendly under specific situations where high N removal is required. The removal rates achieved demonstrated a robust adaptation to influent variations, revealing a removal average of 92% of Biology Oxygen Demand (BOD), 90% of Total Suspended Solids (TSS), 40% of total nitrogen (TN), and 30% of total phosphorus (TP). Moreover, regarding the global warming category, the overall impact was 75% lower compared to other conventional treatments like activated sludge. In conclusion, the LCA revealed that METland® appears as ideal solution for rural areas, considering the low energy requirements and high efficiency to remove organic pollutants, nitrogen, and phosphates from urban wastewater.

Multi-Criteria Evaluation and Sensitivity Analysis for the Optimal Location of Constructed Wetlands (METland) at Oceanic and Mediterranean Areas.

METland is a new variety of Constructed Wetland (CW) for treating wastewater where gravel is replaced by a biocompatible electroconductive material to stimulate the metabolism of electroactive bacteria. The system requires a remarkably low land footprint (0.4 m2/pe) compared to conventional CW, due to the high pollutant removal rate exhibited by such microorganisms. In order to predict the optimal locations for METland, a methodology based on Multi-Criteria Evaluation (MCE) techniques applied to Geographical Information Systems (GIS) has been proposed. Seven criteria were evaluated and weighted in the context of Analytical Hierarchy Process (AHP). Finally, a Global Sensitivity Analysis (GSA) was performed using the Sobol method for resource optimization. The model was tested in two locations, oceanic and Mediterranean, to prove its feasibility in different geographical, demographic and climate conditions. The GSA revealed as conclusion the most influential factors in the model: (i) land use, (ii) distance to population centers, and (iii) distance to river beds. Interestingly, the model could predict best suitable locations by reducing the number of analyzed factors to just such three key factors (responsible for 78% of the output variance). The proposed methodology will help decision-making stakeholders in implementing nature-based solutions, including constructed wetlands, for treating wastewater in rural areas.

Where do we stand to oversee the coronaviruses in aqueous and aerosol environment? Characteristics of transmission and possible curb strategies.

By 17 October 2020, the severe acute respiratory syndrome coronavirus (SARS-CoV-2) has caused confirmed infection of more than 39,000,000 people in 217 countries and territories globally and still continues to grow. As environmental professionals, understanding how SARS-CoV-2 can be transmitted via water and air environment is a concern. We have to be ready for focusing our attention to the prompt diagnosis and potential infection control procedures of the virus in integrated water and air system. This paper reviews the state-of-the-art information from available sources of published papers, newsletters and large number of scientific websites aimed to provide a comprehensive profile on the transmission characteristics of the coronaviruses in water, sludge, and air environment, especially the water and wastewater treatment systems. The review also focused on proposing the possible curb strategies to monitor and eventually cut off the coronaviruses under the authors' knowledge and understanding.

Novel bioelectrochemical strategies for domesticating the electron flow in constructed wetlands.

Constructed wetlands are an effective biofilter-based technology for treating wastewater in a sustainable way; however, their main disadvantage is a large area footprint. To cope with this limitation a new generation of constructed wetlands, the METlands®, have been recently reported. METlands® replace gravel with a granular electrically conductive material to enhance the oxidative metabolisms of electroactive bacteria by facilitating the flux of electron through the material and, consequently, increase bioremediation rates. In this work we evaluated the performance of a new electron sink (e-sink) device with the purpose of controlling and enhancing the electrochemical consumption of electrons from microbial metabolism without energy consumption. The e-sink device was integrated inside the biofilter bed and was tested using different electron acceptors with high redox potentials, like oxygen and hypochlorite. Interestingly, the presence of the e-sink allowed novel redox gradients to form inside the METland® and, consequently, a new electron flow was demonstrated by measuring both the electric potential and current density profiles of the bed. Three independent biofilters were constructed and operated under flooded conditions. Ec-coke and electroconductive biochar (ec-biochar) were used as electrically conductive bed materials, while gravel was used as an inert control. Furthermore, e-sink integration inside the electrically conductive bed outperformed METlands® for removing pollutants, already much more efficient than standard gravel biofilters. COD removal was increased from 90% in METland® to 95% in the e-sink METland® as compared to 75% for the control, while total nitrogen removal was enhanced from 64% in METland® to 71% in e-sink METland® as compared to 55% for the control. Our results indicate that increasing the electrochemical availability of electron acceptors by using the e-sink will be a suitable method for controlling the electron flow inside the filter bed and can be integrated in full scale METlands® for achieving high removal rates.

Microbial electrochemistry for bioremediation.

Lack of suitable electron donors or acceptors is in many cases the key reason for pollutants to persist in the environment. Externally supplementation of electron donors or acceptors is often difficult to control and/or involves chemical additions with limited lifespan, residue formation or other adverse side effects. Microbial electrochemistry has evolved very fast in the past years - this field relates to the study of electrochemical interactions between microorganisms and solid-state electron donors or acceptors. Current can be supplied in such so-called bioelectrochemical systems (BESs) at low voltage to provide or extract electrons in a very precise manner. A plethora of metabolisms can be linked to electrical current now, from metals reductions to denitrification and dechlorination. In this perspective, we provide an overview of the emerging applications of BES and derived technologies towards the bioremediation field and outline how this approach can be game changing.

Electrodes boost microbial metabolism to mineralize antibiotics in manure.

Livestock manures are potential sources of antibiotics in the environment. Sulfamethazine (SMZ), frequently used in veterinary medicine, can enter the environment by using manure as soil fertilizer due to its incomplete absorption in the animal gut and its unmetabolized excretion. The objective of this study was to evaluate the mineralization of 14C-labelled SMZ in manure under a new redox scenario provided by microbial electrochemical reactors, termed microbial electroremediating cells (MERC). These devices aim to overcome the electron acceptor limitation in bacterial oxidative metabolism by means of using electrodes to enhance the biodegradation of pollutants in the environment. Our results revealed that the total degradation of 14C-SMZ reached 43.5% in short term batch laboratory scale experiments under reducing conditions (-400 mV vs. Ag/AgCl). Actually, SMZ mineralization was enhanced up to 10-fold in the early stages (after 2 weeks) in comparison with an electrode-free natural attenuation assay. Moreover, mineralization showed a dependence on electrode potential, with negligible results for conditions set to +400 mV vs Ag/AgCl. The impact of merging electrodes and microorganisms for manure bioremediation suggests a promising future for this emerging technology to treat polluted livestock wastes and prevent soil and groundwater pollution.

Electroactive biofilm-based constructed wetland (EABB-CW): A mesocosm-scale test of an innovative setup for wastewater treatment.

Constructed wetlands (CWs) performance enhancement can be done with intensification strategies. A recent strategy still in study is the coupling with Microbial Electrochemical Technologies (MET). An alternative system using electro-conductive biofilters instead of electrodes and circuits used in MET, resulted in the development of a Microbial Electrochemical-based CW (METland). This system relies on electroactive bacteria (EAB) metabolism to transfer electrons to an electro-conductive material, thus boosting substrate consumption, and diminishing electron availability for biomass build-up and methane generation. In previous studies this biofilters have shown an improvement in biodegradation rates in comparison with subsurface flow CW. However, this set-up is still in development, hence there are uncertainties regarding the dynamics involve in the removal of pollutants. Considering that, this work aimed at establishing the capacity and removal kinetics of organic matter and nutrients in an Electroactive Biofilm-Based CW (EABB-CW). Two electro-conductive materials were tested (PK-A and PK-LSN) in planted and non-planted mesocosms and compared with sand. The systems were operated in a continuous upflow mode for 32 weeks and fed with real wastewater. The electro-conductive systems reached removal efficiencies up to 88% for BOD5, 90% for COD, 46% for NH4-N, and 86% for PO4-P. Organic matter removal in electro-conductive systems was possible even at loading rates 10-fold higher than recommended for horizontal flow CWs. First-order area-based removal constants (k), calculated for organic matter and nutrients are higher than values typically reported for saturated CW and in certain cases comparable with vertical flow CW. The organic removal was correlated with electron current densities measures, as indicator of the presence of EAB. The tested EABB-CW profiles as a promising CW type for the removal of organic matter and PO4-P with margin for modifications to improve nitrogen removal. Future studies with pilot/real scale systems are proposed to validate the findings of this study.

Designing strategies for operating Microbial Electrochemical Systems to clean up polluted soils under non-flooded conditions.

Microbial Electrochemical Systems can be setup in soil either for the harvesting of energy from microbial metabolism (sediment microbial fuel cell, SMFC) or for the bioremediation of polluted environments (Microbial Electroremediating Cell, MERC). However, the precieved need for this technology to be located in flooded environments, to assure the ionic contact between anode and cathode, has limited its implementation in standard soils. This work describes a new configuration that overcomes this limitation by integrating an out-of-soil cathodic chamber with a ceramic barrier so a closed circuit system can be achieved without flooding the soil. On top of harvesting energy under this new configuration we have outperformed natural bioremediation by restoring an atrazine polluted soil as proof of concept. Our results showed that >98% of the initially available atrazine was efficiency removed after 2 weeks with this new MERC configuration, in contrast with a mere 58% removal obtained under natural attenuation. Moreover, a set of toxicological tests using green algae, Salmonella typhimurium and Sorghum saccharatum strongly confirmed a marked lack of toxicity in the soil after the bioelectrochemical treatment in contrast with the natural attenuation that was unable to fulfill the non-toxic level.

Geobacter Dominates the Inner Layers of a Stratified Biofilm on a Fluidized Anode During Brewery Wastewater Treatment.

In this study, we designed a microbial electrochemical fluidized bed reactor (ME-FBR), with an electroconductive anodic bed made of activated carbon particles for treating a brewery wastewater. Under a batch operating mode, acetate and propionate consumption rates were 13-fold and 2.4-fold higher, respectively, when the fluidized anode was polarized (0.2 V) with respect to open circuit conditions. Operating in a continuous mode, this system could effectively treat the brewery effluent at organic loading rates (OLR) over 1.7 kg m-3NRV d-1 and with removal efficiencies of 95 ± 1.4% (hydraulic retention time of 1 day and an influent of 1.7 g-COD L-1). The coulombic efficiency values highly depended upon the OLR applied, and varied from a 56 ± 15% to 10 ± 1%. Fluorescence in situ hybridization (FISH) analysis revealed a relative high abundance of Geobacter species (ca. 20%), and clearly showed a natural microbial stratification. Interestingly, the Geobacter cluster was highly enriched in the innermost layers of the biofilm (thickness of 10 µm), which were in contact with the electroconductive particles of bed, whereas the rest of bacteria were located in the outermost layers. To our knowledge, this is the first time that such a clear microbial stratification has been observed on an anode-respiring biofilm. Our results revealed the relevant role of Geobacter in switching between the electrode and other microbial communities performing metabolic reactions in the outermost environment of the biofilm.

Bioelectroventing: an electrochemical-assisted bioremediation strategy for cleaning-up atrazine-polluted soils.

The absence of suitable terminal electron acceptors (TEA) in soil might limit the oxidative metabolism of environmental microbial populations. Bioelectroventing is a bioelectrochemical strategy that aims to enhance the biodegradation of a pollutant in the environment by overcoming the electron acceptor limitation and maximizing metabolic oxidation. Microbial electroremediating cells (MERCs) are devices that can perform such a bioelectroventing. We also report an overall profile of the 14 C-ATR metabolites and 14 C mass balance in response to the different treatments. The objective of this work was to use MERC principles, under different configurations, to stimulate soil bacteria to achieve the complete biodegradation of the herbicide 14 C-atrazine (ATR) to 14 CO2 in soils. Our study concludes that using electrodes at a positive potential [+600 mV (versus Ag/AgCl)] ATR mineralization was enhanced by 20-fold when compared to natural attenuation in electrode-free controls. Furthermore, ecotoxicological analysis of the soil after the bioelectroventing treatment revealed an effective clean-up in < 20 days. The impact of electrodes on soil bioremediation suggests a promising future for this emerging environmental technology.

Silica immobilization of Geobacter sulfurreducens for constructing ready-to-use artificial bioelectrodes.

Microbial electrochemical technologies (METs) rely on the control of interactions between microorganisms and electronic devices, enabling to transform chemical energy into electricity. We report a new approach to construct ready-to-use artificial bioelectrodes by immobilizing Geobacter sulfurreducens cells in composite materials associating silica gel and carbon felt fibres. Viability test confirmed that the majority of bacteria (ca. 70 ± 5%) survived the encapsulation process in silica and that cell density did not increase in 96 h. The double entrapment within the silica-carbon composite prevented bacterial release from the electrode but allowed a suitable mass transport (ca. 5 min after electron donor pulse), making the electrochemical characterization of the system possible. The artificial bioelectrodes were evaluated in three-electrode reactors and the maximum current displayed was ca. 220 and 150 µA cm-3 using acetate and lactate as electron donors respectively. Cyclic voltammetry of acetate-fed bioelectrodes revealed a sigmoidal catalytic oxidation wave, typical of more advanced-stage biofilms. The presence of G. sulfurreducens within composites was ascertained by SEM analysis, suggesting that only part of the bacterial population was in direct contact with the carbon fibres. Preliminary analyses of the transcriptomic response of immobilized G. sulfurreducens enlightened that encapsulation mainly induces an osmotic stress to the cells. Therefore, ready-to-use artificial bioelectrodes represent a versatile time- and cost-saving strategy for microbial electrochemical systems.