Optimizing electrochemically active microorganisms as a key player in the bioelectrochemical system: Identification methods and pathways to large-scale implementation.

The rapid global economic growth driven by industrialization and population expansion has resulted in significant issues, including reliance on fossil fuels, energy scarcity, water crises, and environmental emissions. To address these issues, bioelectrochemical systems (BES) have emerged as a dual-purpose solution, harnessing electrochemical processes and the capabilities of electrochemically active microorganisms (EAM) to simultaneously recover energy and treat wastewater. This review examines critical performance factors in BES, including inoculum selection, pretreatment methods, electrodes, and operational conditions. Further, authors explore innovative approaches to suppress methanogens and simultaneously enhance the EAM in mixed cultures. Additionally, advanced techniques for detecting EAM are discussed. The rapid detection of EAM facilitates the selection of suitable inoculum sources and optimization of enrichment strategies in BESs. This optimization is essential for facilitating the successful scaling up of BES applications, contributing substantially to the realization of clean energy and sustainable wastewater treatment. This analysis introduces a novel viewpoint by amalgamating contemporary research on the selective enrichment of EAM in mixed cultures. It encompasses identification and detection techniques, along with methodologies tailored for the selective enrichment of EAM, geared explicitly toward upscaling applications in BES.

Biochemical responses and phytoremediation potential of Azolla imbricata (Roxb.) Nakai in water and nutrient media exposed to waste metal cutting fluid along with temperature and humidity stress.

Phytoremediation of metals from water (WM) and nutrient (NM) media exposed to waste metal cutting fluid (WMCF) along with temperature (T) and humidity (H) stress was tested using Azolla imbricata (Roxb.) Nakai. In the absence of WMCF, biomass was higher in NM than in WM during all tests. Surprisingly, opposite results were noted in the presence of WMCF, with growth failing at exposure to > 0.1% and > 0.5% in NM and WM, respectively. Further, correlation analysis of the growth data following WM exposure revealed that biomass was affected positively by T and negatively by H and metal accumulation. Simultaneously, metal accumulation was affected negatively by T and positively by H. The average accumulations of Al, Cd, Cr, Fe, Pb, and Zn across all T/H tests were 540, 282, 71, 1645, 2494 and 1110 mg·kg-1, respectively. The observed bioconcentration factor indicated that A. imbricata acts as a hyperaccumulator or accumulator of Zn (>10) and as either accumulator (>1) or excluder (<1) of the other metals. Overall, the phytoremediation performance of A. imbricata in multi-metal-contaminated WMCF was high in WM under all environmental conditions. Therefore, the use of WM is an economically feasible approach for the removal of metals from WMCF.

Enhanced anaerobic treatment of sulfate-rich wastewater by electrical voltage application.

Treatment of sulfate-rich wastewater with high methane recovery is a major concern due to sulfide inhibition. Here, an electrical voltage (EV) aims to enhance methanogenesis and sulfidogenesis to treat sulfate-rich wastewater. Two (control and EV-applied) reactors were operated with a gradual decrease in chemical oxygen demand (COD)/SO42- ratios (CSR). EV-applied reactor (EVR) demonstrated an increase of ∼30 % in methane production and ∼40 % in sulfate removal, compared to the control till CSR of 2.0. At CSR 1.0, the control failed, while EVR still exhibited a stable performance of 50 % COD-methane recovery. Microbial community results showed that the relative abundance of sulfate-reducing bacteria in EVR was 1.5 times higher than the control. Furthermore, higher relative abundance of dissimilatory sulfate reductase (>50 %) and Ni/Fe hydrogenase (x15) genes demonstrated an improved tolerance against H2S toxicity. This study highlights the importance of EV application by minimizing the byproduct inhibition in sulfate-rich wastewater.

Exploration of deep learning models for real-time monitoring of state and performance of anaerobic digestion with online sensors.

The immediate response to the state disturbances of anaerobic digestion is essential to prevent anaerobic digestion failure. However, frequent monitoring of the state and performance of anaerobic digestion is challenging. Thus, deep learning models were investigated to predict the state and performance variables from online sensor data. The online sensor data, including pH, electric conductivity, and oxidation-reduction potential, were used as the input features to build deep learning models. The state and performance data measured offline were used as the labels. The model performance was compared for several deep learning models of convolutional neural network (CNN), long short-term memory (LSTM), dense layer, and their combinations. The combined model of CNN and bidirectional LSTM was robust and well-generalized in predicting the state and performance variables (R2 = 0.978, root mean square error = 0.031). The combined model is an excellent soft sensor for monitoring the state and performance of anaerobic digestion from electrochemical sensors.

Electrical voltage application as a novel approach for facilitating methanogenic granulation.

Despite having high-rate methanogenic performance, up-flow anaerobic sludge blanket reactor still has challenges regarding long-start up period (3-8 months) for granulation. In this study, "electrical voltage (EV, 0.3 V) application" was attempted for facilitating granulation in the continuous operation with increased organic loading rates (0.5-11.0 kg COD/m3/d). Up to 11.0 kg COD/m3/d, EV-reactor exhibited the stable performance, while the control failed. After 49 days of operation (at 7 kg COD/m3/d), the granules collected from EV-reactor had larger diameter (2.3 vs 1.6 mm), higher settling velocity (2.6 vs 1.9 cm/s), and higher hydrophobicity (52.1 % vs 34.5 %), compared to the control. EV application also increased the specific methanogenic activity for propionate and hydrogen almost by two times. The relative abundance of Pseudomonas sp. (quorum sensing (QS)-related microbe) in EV-reactor was 17 % higher than that in the control. In addition, EV application increased the expression of QS genes significantly by 27 times.

Electrostatic Fields Promote Methanogenesis More than Polarized Bioelectrodes in Anaerobic Reactors with Conductive Materials.

Direct interspecies electron transfer (DIET) is a breakthrough that can surpass the limitations of anaerobic digestion. Conductive materials and polarized bioelectrodes are known to induce DIET for methane production but are still challenging to apply at a field scale. Herein, compared to polarized bioelectrodes, electrostatic fields that promote DIET were investigated in an anaerobic reactor with conductive materials. As a conductive material, activated carbon enriched its surface with electroactive microorganisms to induce DIET (cDIET). cDIET improved the methane yield to 254.6 mL/g CODr, compared to the control. However, polarized bioelectrodes induced electrode-mediated DIET and biological DIET (bDIET), in addition to cDIET, improving the methane yield to 310.7 mL/g CODr. Electrostatic fields selectively promoted bDIET and cDIET for further methane production compared to the polarized bioelectrodes. As the contribution of DIET increased, the methane yield increased, and the substrate residue decreased, resulting in a significant improvement in methane production.

External electric field promotes ammonia stripping from wastewater.

The gas stripping process is widely used for the removal and recovery of ammonia from wastewater. The ammonia removal in the stripping process depends on the pH, temperature, and air supply, and in general, 10.5, 60 °C, 5 L/min or more are recommended as near-optimal. However, alkaline chemicals and energy can seriously burden the stripping process operation, depending on the wastewater characteristics. Herein, external electric field-coupled ammonia stripping, which improves ammonia removal from aqueous solutions, was investigated. The ammonia removal in the conventional stripping was between 17.7 % and 90.6 %, depending on how close to the near-optimal conditions. The electric field increased the removal efficiency from 51.1 % to 94.3 %, as the strength and frequency increased to 15 V/cm and 50 MHz. The electric field promotion of ammonia stripping correlated closely with the increase in ammonia molecular diffusion. The electric field has been shown to improve the diffusion of ammonia molecules without dielectric heating by increasing the kinetic energy. The electric field improved the ammonia removal more as the pH, temperature, and air supply conditions were far from optimal. The electric field reduces the alkaline chemicals and the electric energy for heating and air supply, and the energy consumption to form the electric field is only a few watts. The electric field-coupled process offers a new gas stripping platform that can economically improve ammonia removal from wastewater.

Treatment of phenol wastewater using nitrogen-doped magnetic mesoporous hollow carbon.

Nitrogen-doped magnetic mesoporous hollow carbon (NMMHC) was prepared to realize effective adsorption of phenol from wastewater. The chemical and physical properties of NMMHC were analyzed, and the effects of adsorption time, initial pH, and phenol concentration on the adsorption capacity of NMMHC were studied. Adsorption kinetics and isotherm models were used to explain the adsorption properties. The results showed that the specific surface area, type of nitrogen group, and nitrogen content of NMMHC are related to the carbonization temperature. Chemical interaction was demonstrated to be present in the process of adsorption, which was characterized as monolayer adsorption. In addition, the adsorption mechanism was studied by attenuated total internal reflectance Fourier transform infrared spectroscopy and analysis of non-covalent interactions. This study found that non-covalent interactions between NMMHC and phenol molecules are van der Waals interactions, and nitrogen-containing groups increase the phenol adsorption capacity by enhancing such interactions. The π-π interactions between the nitrogen groups and phenol molecules also enhanced the adsorption energy.

Electro-fermentation for biofuels and biochemicals production: Current status and future directions.

Electro-fermentation is an emerging bioporcess that could regulate the metabolism of electrochemically active microorganisms. The provision of electrodes for the fermentation process that functions as an electron acceptor and supports the formation and transportation of electrons and protons, consequently producing bioelectricity and value-added chemicals. The traditional method of fermentation has several limitations in usability and economic feasibility. Subsequently, a series of metabolic processes occurring in conventional fermentation processes are most often redox misaligned. In this regard, electro-fermentation emerged as a hybrid technology which can regulate a series of metabolic processes occurring in a bioreactor by regulating the redox instabilities and boosting the overall metabolic process towards high biomass yield and enhanced product formation. The present article deals with microorganisms-electrode interactions, various types of electro-fermentation systems, comparative evaluation of pure and mixed culture electro-fermentation application, and value-added fuels and chemical synthesis.

Influence of electrostatic field and conductive material on the direct interspecies electron transfer for methane production.

The influence of electrostatic field on the direct interspecies electron transfer (DIET) pathways for methane production was investigated in a batch bioelectrochemical anaerobic digester (BEAD). The ultimate methane production and methane yield in the BEAD reactor saturated to 925 ± 29 mL/L and 309.9 ± 9.6 mL CH4/g COD, respectively, which were much higher than 616 ± 3 mL/L and 205.4 ± 205.4 mL CH4/g COD in the anaerobic digester (AD). In the cyclic voltammogram (CV) for bulk solution, the oxidation peak current was 0.52 mA in the BEAD reactor, which was higher than 0.24 mA of AD reactor. This shows that the oxidizing ability of microorganisms was greatly improved in the BEAD reactor. Anaerolineaceae, a well-known electroactive bacterial family, was well enriched in the BEAD reactor. It indicates that the electrostatic field can enrich the electroactive bacteria and activate the DIET pathways for methane production. Moreover, the conductive material (activated carbon) further improved the performance of BEAD reactor, implies that the conductivities of bulk solution is one of the important parameters for the DIET pathways.

Enhanced Anaerobic Digestion of Long Chain Fatty Acid by Adding Magnetite and Carbon Nanotubes.

This study investigated the impact of stimulating direct interspecies electron transfer (DIET), by supplementing nano-sized magnetite (nFe3O4, 0.5 g Fe/g VSS) and carbon nanotubes (CNT, 1 g/L), in anaerobic digestion of oleic acid (OA) at various concentrations (0.10 - 4.00 g chemical oxygen demand(COD)/L). Both supplementations could enhance CH4 production, and its beneficial impact increased with increased OA concentration. The biggest improvements of 114% and 165% compared to the control were achieved by nFe3O4 and CNT, respectively, at OA of 4 g COD/L. The enhancement can be attributed to the increased sludge conductivity: 7.1 ± 0.5 (control), 12.5 ± 0.8 (nFe3O4-added), and 15.7 ± 1.1 µS/cm (CNT-supplemented). Dissolved iron concentration, released from nFe3O4, seemed to have a negligible role in improving CH4 production. The excretion of electron shuttles, i.e., humic-like substances and protein-like substances, were found to be stimulated by supplementing nFe3O4 and CNT. Microbial diversity was found to be simplified under DIET-stimulating conditions, whereby five genera accounted for 88% of the total sequences in the control, while more than 82% were represented by only two genera (Methanotrix concilli and Methanosarcina flavescens) by supplementing nFe3O4 and CNT. In addition, the abudance of electro-active bacteria such as Syntrophomonas zehnderi was significantly increased from 17% to around 45%.

Nitrite and nitrate as electron acceptors for bioelectrochemical ammonium oxidation under electrostatic field.

Bioelectrochemical ammonium oxidation with nitrite and nitrate as electron acceptors was investigated in bulk solution exposed to electrostatic field. In a bioelectrochemical reactor, electroactive nitrogen removal bacteria including ammonium oxidizing exoelectrogens (AOE) and denitrifying electrotrophs (DNE) were enriched by electrostatic field of 0.2 V/cm in a bulk solution containing nitrite, nitrate, and ammonium. Ammonium was oxidized simultaneously with decreases in nitrite and nitrate as electron acceptors due to direct interspecies electron transfer between AOE and DNE. The specific ammonium oxidation rate was 48 mg NH4-N/g VSS.d when nitrate fraction was 1/3 in the electron acceptor composed of nitrite and nitrate. The specific ammonium oxidation rate gradually decreased with increasing nitrate fraction. However, it was still 24 mg NH4-N/g VSS.d when nitrate was the only electron acceptor. This indicates that nitrate can be used as an electron acceptor for bioelectrochemical ammonium oxidation, although it is a less effective than nitrite. This finding provides an advantage that strict nitritation which selectively produces nitrite from ammonium can be avoided when treating ammonia-rich wastewater in a bioelectrochemical reactor.

Rapid detection of heavy metal-induced toxicity in water using a fed-batch sulfur-oxidizing bacteria (SOB) bioreactor.

A fed-batch bioreactor based on sulfur-oxidizing bacteria (SOB) was tested for rapid detection of heavy metal-induced toxicity in water. For this evaluation, SOB were exposed to water contaminated by selenium, mercury, hexavalent chromium, arsenic, cyanide, cadmium, and lead for 2 h and their inhibition rates were analyzed based on changes in electrical conductivity (EC). The results demonstrate that SOB were highly inhibited by selenium, mercury, hexavalent chromium, and arsenic but not by cyanide, cadmium, and lead. The 2 h half maximum effective concentrations (EC50) of SOB for selenium, mercury, hexavalent chromium, and arsenic were estimated to be 0.33, 0.89, 1.18, and 0.24 mg/L, respectively, which are comparable or lower than earlier reports in the literature. However, the EC50 or EC20 values of SOB for cyanide, cadmium, and lead were notably higher compared to findings from previous toxicity tests that employed other microorganisms. The findings from the current study suggest that the fed-batch SOB bioreactor is suitable for rapid detection of toxicity induced by selenium, mercury, hexavalent chromium, and arsenic in water.

Electroactive microorganisms enriched from activated sludge remove nitrogen in bioelectrochemical reactor.

The bioelectrochemical anaerobic nitrogen removal was demonstrated in an anaerobic batch reactor equipped with a pair of polarized bioelectrodes. The bioelectrochemical reactor was operated in sequential batch mode after inoculating activated sludge and polarizing the electrode to 0.6 V. The medium contains ammonium, nitrite, alkalinity and trace minerals, but no organic carbon source. By the repetitive sequential operation, simultaneous removals of ammonium, nitrite and alkalinity were improved, and the electrochemical activity of the bulk sludge was confirmed from the redox peaks of the cyclic voltammogram. This indicates that ammonia oxidizing exoelectrogens (AOE) and denitritating electrotrophs (DNE) were enriched more in the bulk solution. Biogas production that mainly consisted of nitrogen was observed from the bioelectrochemical reactor, and the minor components in the biogas were methane and carbon dioxide. This demonstrates that AOE use nitrite as an electron acceptor to oxidize ammonia. The requirements of nitrite and alkalinity for the removal of ammonia nitrogen are around 0.72 mg NO2-N/mg NH4-N and 1.73 mg as CaCO3/mg NH4-N, respectively, and nitrate was not produced as a by-product. The bacterial groups involved in the bioelectrochemical nitrogen removal are electroactive autotrophs and can be enriched from activated sludge by polarized electrode. This bioelectrochemical ammonia oxidation is a novel approach recommended for treatment of nitrogen-rich wastewater.

Modeling methane production in anaerobic forward osmosis bioreactor using a modified anaerobic digestion model No. 1.

Anaerobic membrane bioreactor (AnMBR) using microfiltration (MF) or ultrafiltration (UF) membranes was introduced to enhance poor biomass retention of conventional anaerobic digestion (CAD). Recently, forward osmosis (FO) membrane have been applied to AnMBR, which is called AnFOMBR. FO membrane assures not only high biomass retention but also high removal efficiency for low molecular weight (LMW) matters. Methane production rates in CAD, AnMBR, and AnFOMBR were compared using a modified IWA anaerobic digestion model No. 1 (ADM1) in this work. Accumulation of biomass in AnMBR/AnFOMBR results in enhanced biochemical reaction and gains more methane production. AnFOMBR may experience a significant inhibition by accumulated free ammonia and cations, although concentrated soluble substrates rejected by FO membrane are favorable for more methane production. Rejection rate of inorganic nitrogen is a key parameter to determine the inhibition in methane production of AnFOMBR.

Polarized electrode enhances biological direct interspecies electron transfer for methane production in upflow anaerobic bioelectrochemical reactor.

The influence of polarized electrodes on the methane production, which depends on the sludge concentration, was investigated in upflow anaerobic bioelectrochemical (UABE) reactor. When the polarized electrode was placed in the bottom zone with a high sludge concentration, the methane production was 5.34 L/L.d, which was 53% higher than upflow anaerobic sludge blanket (UASB) reactor. However, the methane production was reduced to 4.34 L/L.d by placing the electrode in the upper zone of the UABE reactor with lower sludge concentration. In the UABE reactor, the methane production was mainly improved by the enhanced biological direct interspecies electron transfer (bDIET) pathway, and the methane production via the electrode was a minor fraction of less than 4% of total methane production. The polarized electrodes that placed in the bottom zone with a high sludge concentration enhance the bDIET for methane production in the UABE reactor and greatly improve the methane production.

Electroactive microorganisms in bulk solution contribute significantly to methane production in bioelectrochemical anaerobic reactor.

The role of anaerobic microorganisms suspended in the bulk solution on methane production was investigated in a bioelectrochemical anaerobic reactor with the electrode polarized at 0.5 V. The electron transfer from substrate to methane and hydrogen were 25% and 7.5%, respectively, in the absence of the anaerobic microorganisms in the bulk solution. As the anaerobic microorganisms increased to 4400 mg/L, the electrons transferred to methane increased to 83.3% but decreased to 0.3% in hydrogen. The electroactive microorganisms (EAM), including exoelectrogens and electrotrophs, enriched in the bulk solution that confirmed by the redox peaks in the cyclic voltammogram was proportional to the anaerobic microorganism. The methane yield based on COD removal was dependent on the anaerobic microorganisms in the bulk solution rather than on the bioelectrode surface. The EAM suspended in the bulk solution are enriched by the polarized electrode, and significantly improve methane production in bioelectrochemical anaerobic reactor.

Influence of conductive material on the bioelectrochemical removal of organic matter and nitrogen from low strength wastewater.

The treatment of low strength wastewater that has the level of discharge standard for wastewater treatment plant was studied using an upflow bioelectrochemical reactor with an applied voltage of 0.6 V. The direct interspecies electron transfer (DIET) between electroactive bacteria was activated in the upflow bioelectrochemical reactor, which improved the substrate affinity of bacteria. The effluent qualities in COD and ammonia nitrogen was stable at less than 3.5 mg/L and 7.46 mg/L at 1 h of hydraulic retention time, respectively. The conductive materials, including conductive sheets and conductive particles, further increased the biomass retention and the DIET by altering the abundance of dominant bacterial groups. The effluent qualities in COD and ammonia nitrogen was improved up to 1.98 mg/L and 2.65 mg/L, respectively, by the conductive sheets. The upflow bioelectrochemical reactor with conductive materials is a good tertiary treatment process for improving the quality of the final effluent discharged from wastewater treatment plant.

Bioelectrochemical enhancement of direct interspecies electron transfer in upflow anaerobic reactor with effluent recirculation for acidic distillery wastewater.

Methane production in the upflow anaerobic bioelectrochemical reactor (UABE) treating acidic distillery wastewater was compared to the upflow anaerobic sludge blanket reactor (UASB), and the electron transfer pathways for methane production were also evaluated in the effluent recirculation. The methane productions from reactors were influenced by the low pH of influent wastewater. However, the methane production rate and yield of the UABE were 2.08L/L.d and 320mL/g CODr, which were higher than the UASB. The effluent recirculation containing alkalinity neutralized the acidic influent and increased the upflow velocity in both reactors, and improved the direct interspecies electron transfer more in the UABE. When the effluent recirculation ratio was 3.0 in the UABE, the methane production rate and yield were reached up to 3.88L/L.d and 501.0mL/g CODr, respectively. The UABE requires electrode installation and electrical energy for operation, but the benefits from increased methane production are much higher.

Bioremediation of contaminated coastal sediment: Optimization of slow release biostimulant ball using response surface methodology (RSM) and stabilization of metals from contaminated sediment.

The aim of the present study is to optimize the slow release biostimulant ball (BSB) for bioremediation of contaminated coastal sediment using response surface methodology (RSM). Metals contamination and stabilization of metals in coastal sediments using BSB were investigated. The effects of BSB size (1-5cm), distance (1-10cm), and time (1-4months) on the stabilization of metals including Fe, Cd, Cu, and Pb were determined. The maximum stabilization percentages of Fe, Cd, Cu, and Pb, of 64.5%, 54.9%, 63.8%, and 47.6%, respectively, were observed at a 3cm ball size, 5.5cm distance, and a period of 4months; these values are the optimum conditions for effective treatment of contaminated coastal sediment. The determination coefficient of the R2 value suggests that >91.55%, 89.97%, 96.10%, and 86.40% of the variance is attributable to the variables of Fe, Cd, Cu, and Pb, respectively.