Microbial electrochemical Cr(VI) reduction in a soil continuous flow system

Microbial electrochemical technologies represent innovative approaches to contaminated soil and groundwater remediation and provide a flexible framework for removing organic and inorganic contaminants by integrating electrochemical and biological techniques. To simulate in situ microbial electrochemical treatment of groundwater plumes, this study investigates Cr(VI) reduction within a bioelectrochemical continuous flow (BECF) system equipped with soil-buried electrodes, comparing it to abiotic and open-circuit controls. Continuous-flow systems were tested with two chromium-contaminated solutions (20-50 mg Cr(VI)/L). Additional nutrients, buffers, or organic substrates were introduced during the tests in the systems. With an initial Cr(VI) concentration of 20 mg/L, 1.00 mg Cr(VI)/(L day) bioelectrochemical removal rate in the BECF system was observed, corresponding to 99.5% removal within nine days. At the end of the test with 50 mg Cr(VI)/L (156 days), the residual Cr(VI) dissolved concentration was two orders of magnitude lower than that in the open circuit control, achieving 99.9% bioelectrochemical removal in the BECF. Bacteria belonging to the orders Solirubrobacteriales, Gaiellales, Bacillales, Gemmatimonadales, and Propionibacteriales characterized the bacterial communities identified in soil samples; differently, Burkholderiales, Mycobacteriales, Cytophagales, Rhizobiales, and Caulobacterales characterized the planktonic bacterial communities. The complexity of the microbial community structure suggests the involvement of different microorganisms and strategies in the bioelectrochemical removal of chromium. In the absence of organic carbon, microbial electrochemical removal of hexavalent chromium was found to be the most efficient way to remove Cr(VI), and it may represent an innovative and sustainable approach for soil and groundwater remediation. Integr Environ Assess Manag 2024;00:1-17. © 2024 The Author(s). Integrated Environmental Assessment and Management published by Wiley Periodicals LLC on behalf of Society of Environmental Toxicology & Chemistry (SETAC)

Performance Analysis and Microbial Community Evolution of In Situ Biological Biogas Upgrading with Increasing H2/CO2 Ratio

The effect of the amount of hydrogen supplied for the in situ biological biogas upgrading was investigated by monitoring the process and evolution of the microbial community. Two parallel reactors, operated at 37°C for 211 days, were continuously fed with sewage sludge at a constant organic loading rate of 1.5 gCOD∙(L∙d)-1 and hydrogen (H2). The molar ratio of H2/CO2 was progressively increased from 0.5 : 1 to 7 : 1 to convert carbon dioxide (CO2) into biomethane via hydrogenotrophic methanogenesis. Changes in the biogas composition become statistically different above the stoichiometric H2/CO2 ratio (4 : 1). At a H2/CO2 ratio of 7 : 1, the methane content in the biogas reached 90%, without adversely affecting degradation of the organic matter. The possibility of selecting, adapting, and enriching the original biomass with target-oriented microorganisms able to biologically convert CO2 into methane was verified: high throughput sequencing of 16S rRNA gene revealed that hydrogenotrophic methanogens, belonging to Methanolinea and Methanobacterium genera, were dominant. Based on the outcomes of this study, further optimization and engineering of this process is feasible and needed as a means to boost energy recovery from sludge treatment

Microbial Assisted Hexavalent Chromium Removal in Bioelectrochemical Systems

Groundwater is the environmental matrix that is most frequently affected by anthropogenic hexavalent chromium contamination. Due to its carcinogenicity, Cr(VI) has to be removed, using environmental-friendly and economically sustainable remediation technologies. BioElectrochemical Systems (BESs), applied to bioremediation, thereby offering a promising alternative to traditional bioremediation techniques, without affecting the natural groundwater conditions. Some bacterial families are capable of oxidizing and/or reducing a solid electrode obtaining an energetic advantage for their own growth. In the present study, we assessed the possibility of stimulating bioelectrochemical reduction of Cr(VI) in a dual-chamber polarized system using an electrode as the sole energy source. To develop an electroactive microbial community three electrodes were, at first, inserted into the anodic compartment of a dual-chamber microbial fuel cell, and inoculated with sludge from an anaerobic digester. After a period of acclimation, one electrode was transferred into a polarized system and it was fixed at −0.3 V (versus standard hydrogen electrode, SHE), to promote the reduction of 1000 µg Cr(VI) L−1. A second electrode, served for the set-up of an open circuit control, operated in parallel. Cr(VI) dissolved concentration was analysed at the initial, during the experiment and final time by spectrophotometric method. Initial and final microbial characterization of the communities enriched in polarized system and open circuit control was performed by 16S rRNA gene sequencing. The bioelectrode set at −0.3 V showed high Cr(VI) removal efficiency (up to 93%) and about 150 µg L−1 day−1 removal rate. Similar efficiency was observed in the open circuit (OC) even at about half rate. Whereas, purely electrochemical reduction, limited to 35%, due to neutral operating conditions. These results suggest that bioelectrochemical Cr(VI) removal by polarized electrode offers a promising new and sustainable approach to the treatment of groundwater Cr(VI) plumes, deserving further research

Enhanced Exoelectrogenic Activity of <i>Cupriavidus metallidurans</i> in Bioelectrochemical Systems through the Expression of a Constitutively Active Diguanylate Cyclase

Electroactive bacteria have a wide range of applications, including electricity production, bioremediation, and the sensing of toxic compounds. Bacterial biofilm formation is often mediated by the second messenger cyclic guanosine monophosphate (c-di-GMP) synthesized by a diguanylate cyclase (DGC). The role of c-di-GMP in the expression of c-type cytochromes has been previously reported. The aim of this study was to determine the bioelectrogenic activity of Cupriavidus metallidurans strain CH34 pJBpleD*, which possesses a constitutively active DGC that increases c-di-GMP levels. Notably, the heterologous expression of the constitutively active DGC in C. metallidurans strain CH34 pJBpleD* showed a higher biofilm formation and increased the electrical current production up to 560%. In addition, C. metallidurans CH34 pJBpleD* showed increased levels of c-type cytochrome-associated transcripts compared with the wild-type strain CH34. Scanning electron microscopies revealed a denser extracellular matrix with an increased exopolymeric substance content in the CH34 pJBpleD* biofilm on the electrode surface. The results of this study suggest that higher levels of c-di-GMP synthesized by a constitutively active diguanylate cyclase in C. metallidurans strain CH34 pJBpleD* activated the formation of an electroactive biofilm on the electrode, enhancing its exoelectrogenic activity

Progress Towards Bioelectrochemical Remediation of Hexavalent Chromium

Chromium is one of the most frequently used metal contaminants. Its hexavalent form Cr(VI), which is exploited in many industrial activities, is highly toxic, is water-soluble in the full pH range, and is a major threat to groundwater resources. Alongside traditional approaches to Cr(VI) treatment based on physical-chemical methods, technologies exploiting the ability of several microorganisms to reduce toxic and mobile Cr(VI) to the less toxic and stable Cr(III) form have been developed to improve the cost-effectiveness and sustainability of remediating hexavalent chromium-contaminated groundwater. Bioelectrochemical systems (BESs), principally investigated for wastewater treatment, may represent an innovative option for groundwater remediation. By using electrodes as virtually inexhaustible electron donors and acceptors to promote microbial oxidation-reduction reactions, in in situ remediation, BESs may offer the advantage of limited energy and chemicals requirements in comparison to other bioremediation technologies, which rely on external supplies of limiting inorganic nutrients and electron acceptors or donors to ensure proper conditions for microbial activity. Electron transfer is continuously promoted/controlled in terms of current or voltage application between the electrodes, close to which electrochemically active microorganisms are located. Therefore, this enhances the options of process real-time monitoring and control, which are often limited in in situ treatment schemes. This paper reviews research with BESs for treating chromium-contaminated wastewater, by focusing on the perspectives for Cr(VI) bioelectrochemical remediation and open research issues

Enhanced Exoelectrogenic Activity of Cupriavidus metallidurans in Bioelectrochemical Systems through the Expression of a Constitutively Active Diguanylate Cyclase

Electroactive bacteria have a wide range of applications, including electricity production, bioremediation, and the sensing of toxic compounds. Bacterial biofilm formation is often mediated by the second messenger cyclic guanosine monophosphate (c-di-GMP) synthesized by a diguanylate cyclase (DGC). The role of c-di-GMP in the expression of c-type cytochromes has been previously reported. The aim of this study was to determine the bioelectrogenic activity of Cupriavidus metallidurans strain CH34 pJBpleD*, which possesses a constitutively active DGC that increases c-di-GMP levels. Notably, the heterologous expression of the constitutively active DGC in C. metallidurans strain CH34 pJBpleD* showed a higher biofilm formation and increased the electrical current production up to 560%. In addition, C. metallidurans CH34 pJBpleD* showed increased levels of c-type cytochrome-associated transcripts compared with the wild-type strain CH34. Scanning electron microscopies revealed a denser extracellular matrix with an increased exopolymeric substance content in the CH34 pJBpleD* biofilm on the electrode surface. The results of this study suggest that higher levels of c-di-GMP synthesized by a constitutively active diguanylate cyclase in C. metallidurans strain CH34 pJBpleD* activated the formation of an electroactive biofilm on the electrode, enhancing its exoelectrogenic activity

Anaerobic electrogenic oxidation of toluene in a continuous-flow bioelectrochemical reactor: Process performance, microbial community analysis, and biodegradation pathways

Microbial electrochemical systems (MES) represent an innovative reagent-free technology for in situ remediation of groundwater contaminated by petroleum hydrocarbons. Here we describe the long-term (>160 days) anaerobic treatment of synthetic groundwater containing toluene (25 mg L-1) in a novel laboratory-scale, continuous-flow bioelectrochemical reactor, termed the "bioelectric well". Under optimal operating conditions (i.e., anode potential potentiostatically controlled at +0.2 V vs. SHE and recycle flow-rate set at 75 mL min-1), the observed electrogenic toluene oxidation rate was 67.2 ± 5.7 mg L d-1, a value which is among the highest reported in the literature for laboratory-scale anaerobic treatment systems. Correspondingly, electric current was 5.1 ± 0.1 mA and the Coulombic efficiency (i.e., the yield of toluene conversion into electric current) was 79 ± 7%. Electrogenic toluene oxidation was most likely catalyzed by Geobacter species, which were found to dominate the surface of the graphite anode. The combined application of GC-MS (for detection and identification of metabolites) and qPCR (for quantification of functional genes) revealed that toluene degradation was initiated by fumarate addition, an activation pathway that is widely distributed among anaerobic hydrocarbon-degrading microorganisms. Collectively, the results of this study indicate that the bioelectric well holds remarkable potential for in situ treatment of groundwater contaminated by petroleum hydrocarbons

Bioelectrochemical treatment of groundwater containing BTEX in a continuous-flow system: substrate interactions, microbial community analysis, and impact of sulfate as a co-contaminant

Summarization: Microbial electrochemical technologies (MET) are increasingly being considered for in situ remediation of contaminated groundwater. However, their application potential for the simultaneous treatment of complex mixtures of organic and inorganic contaminants, has been only marginally explored. Here we have analyzed the performance of the ‘bioelectric well’, a previously developed bioelectrochemical reactor configuration, in the treatment of benzene, toluene, ethyl-benzene and xylenes (BTEX) mixtures. Although to different extents, all BTEX were found to be degraded in the bioelectrochemical system, operated using a continuous-flow of groundwater at a hydraulic retention time of 8.8 h, with the graphite anode potentiostatically controlled at +0.200 V vs. the standard hydrogen electrode. In the case of toluene and ethyl-benzene, biodegradation was further confirmed by the GC–MS identification of fumarate-addition metabolites, previously shown to be involved in the activation of these contaminants under anaerobic conditions. Degradation rates were higher for toluene (31.3 ± 1.5 mg/L d) and lower for benzene (6.1 ± 0.3 mg/L d), ethyl-benzene (3.3 ± 0.1 mg/L d), and xylenes (4.5 ± 0.2 mg/L d). BTEX degradation was linked to electric current generation, with coulombic efficiencies falling in the range 53–69%, although methanogenesis also contributed to contaminant degradation. Remarkably, the system also allowed removal of sulfate simultaneously with toluene. Sulfate removal was likely driven by the hydrogen abiotically generated at the cathode. Taken as a whole, these findings highlight the remarkable potential of this innovative reactor configuration for application in a variety of contamination scenarios.Παρουσιάστηκε στο: New Biotechnolog

Characterization of the microbial community in ripened Pecorino Toscano cheese affected by pink discoloration

Pink discoloration defect can cause economic losses for cheese producers due to the impossibility to sell the defected cheese, but few knowledge is currently available on the causes of this defect. To gain more insight on the causes that lead to the formation of pink discoloration in Pecorino Toscano cheese with the Protected Designation of Origin (PDO) status, the bacterial community in defected and not defected cheese was characterized by high-throughput sequencing of bacterial 16S rRNA gene. The bacterial community in the defected cheese significantly differed compared to the control. The relative abundance of the genera Acidipropionibacterium, Enterococcus, Escherichia/Shigella, Lactobacillus, Lentilactobacillus and Propionibacterium was higher in the cheese with pink discoloration defect. The concentration of short chain fatty acids and of lactic acid in cheese was measured and a shift towards the production of propionate in the cheese with pink discoloration defect was observed. Furthermore, the possible involvement of microbially produced vitamin B12 in the formation of pink discoloration was not supported by the data, since a tendency to a lower concentration of vitamin B12 was measured in the defected cheese compared to the control