Electroactive microorganisms (EAMs) play a vital role in biogeochemical cycles by facilitating extracellular electron transfer. They demonstrate remarkable adaptability to river sediments that are characterized by pollution and poor water quality, significantly contributing to the sustainability of river ecosystems. However, the distribution and diversity of EAMs remain poorly understood. In this study, 16S rRNA gene high-throughput sequencing and real-time fluorescence quantitative PCR were used to assess EAMs in 160 samples collected from eight rivers within the Pearl River Delta of Southern China. The results indicated that specialized EAMs communities in polluted sediments exhibited variations in response to water quality and sediment depth. Compared to clean sediment, polluted sediments showed a 4.5% increase in the relative abundances of EAMs communities (59 genera), with 45- and 17-times higher abundances of Geobacter and cable bacteria. Additionally, the abundance of cable bacteria decreased with increasing sediment depth in polluted sediments, while the abundance of L. varians GY32 exhibited an opposite trend. Finally, the abundances of Geobacter, cable bacteria, and L. varians GY32 were positively correlated with the abundance of filamentous microorganisms (FMs) across all samples, with stronger interactions in polluted sediments. These findings suggest that EAMs demonstrate heightened sensitivity to polluted environments, particularly at the genus (species) level, and exhibit strong adaptability to conditions characterized by high levels of acid volatile sulfide, low dissolved oxygen, and elevated nitrate nitrogen. Therefore, environmental factors could be manipulated to optimize the growth and efficiency of EAMs for environmental engineering and natural restoration applications.

Electrobioremediation is one of the most innovative disciplines for treating organic pollutants and it is based on the ability of electroactive bacteria to exchange electrons with electroconductive materials. Electroactive biofilters have been demonstrated to be efficient for treating urban wastewater with a low footprint; however, their application can be expanded for treating industrial wastewater containing significant concentrations (2.4 %vol) of commercial surfactants (containing lauryl sulfate, lauryl ether sulfate, cocamydopropyl betaine, and dodecylbenzene sulfonate, among others). Our electroactive biofilter outperformed a conventional inert biofilter made of gravel for all tested conditions, reaching removal rates as high as 4.5 kg COD/m3bed·day and withstood Organic Loading Rates as high as 9 Kg COD/m3·d without significantly affecting removal efficiency. The biomass accumulation reduced available bed volume in the electroactive biofilter just by 39 %, while the gravel biofilter decreased by 80 %. Regarding microbial communities, anaerobic and electroactive bacteria represented a substantial proportion of the total population in the electroactive biofilter. Pseudomonas was the dominant genus, while Cupriavidus, Shewanella, Citrobacter, Desulfovibrio, and Arcobacter were potential electroactive strains found in relevant proportions. The microbial community\'s composition might be the key to understanding how high removal rates can coexist with limited biomass production, making electroactive biofilters a promising strategy to overcome classical biofilter limitations.

By integrating electroactive genes into engineered sensing microorganisms, information about the object to be measured can be converted into the output of an electrical signal, omitting the process of converting the output of an electrical signal in conventional sensing strategies and simplifying the steps of biosensor development. By utilizing synthetic biology methods, we can not only create novel genetic circuits by using logic gate operations and integrating genes from other biological components, solving biosensing issues in living systems and enhancing sensor performance, but also convert various types of genetic circuits into electrical signals, broadening the application range of biosensors. Here, we describe an example of how to genetically engineer microorganisms with electroactive genes and the fabrication of an electrochemical microbial biosensor.

Microorganisms are key players in the global biogeochemical sulfur cycle. Among them, some have garnered particular attention due to their electrical activity and ability to perform extracellular electron transfer. A growing body of research has highlighted their extensive phylogenetic and metabolic diversity, revealing their crucial roles in ecological processes. In this review, we delve into the electron transfer process between sulfate-reducing bacteria and anaerobic alkane-oxidizing archaea, which facilitates growth within syntrophic communities. Furthermore, we review the phenomenon of long-distance electron transfer and potential extracellular electron transfer in multicellular filamentous sulfur-oxidizing bacteria. These bacteria, with their vast application prospects and ecological significance, play a pivotal role in various ecological processes. Subsequently, we discuss the important role of the pili/cytochrome for electron transfer and presented cutting-edge approaches for exploring and studying electroactive microorganisms. This review provides a comprehensive overview of electroactive microorganisms participating in the biogeochemical sulfur cycle. By examining their electron transfer mechanisms, and the potential ecological and applied implications, we offer novel insights into microbial sulfur metabolism, thereby advancing applications in the development of sustainable bioelectronics materials and bioremediation technologies.

A biological treatment is the core process for removing organic pollutants from industrial wastewater. However, industrial wastewater often contains large amounts of toxic and harmful pollutants, which can inhibit the activity of microorganisms in a treatment system, precipitate the deterioration of effluent quality, and threaten water ecological security from time to time. In most of the existing anaerobic biological treatment processes, toxic effects on microorganisms are determined according to the amounts of end-products of the biochemical reactions, and the evaluation results are relatively lacking. When microorganisms contact toxic substances, changes in biological metabolic activity precede the accumulation of reaction products. As sensitive units, electroactive microorganisms can generate electrical signals, a change in which can directly reflect the toxicity level. The applications of electroactive microorganisms for the toxicity monitoring of wastewater are very promising. Further attention needs to be paid to considering the appropriate evaluation index, the influence of the environment on test results, mechanisms, and other aspects. Therefore, we reviewed the literature regarding the above aspects in order to provide a research foundation for the practical application of electroactive microorganisms in toxicant monitoring.

The worldwide issue of nitrate-contaminated groundwater requires practical solutions, and electro-bioremediation offers a promising and sustainable treatment. While it has shown potential benefits, there is room for improvement in treatment rates, which is crucial for its further and effective implementation. In this field, electrochemical characterisation is a valuable tool for providing the foundation for optimising bioelectrochemical reactors, but applying it in fixed-bed reactors is challenging due to its high intrinsic electrical resistance. To overcome these challenges, this study employed the easy and swift eClamp methodology to screen different process parameters and their influence on the performance of fixed-bed denitrifying biocathodes composed of granular graphite. Granules were extracted and studied ex-situ under controlled conditions while varying key operational parameters (such as pH, temperature, and nitrate concentration). In the studied biocathode, the extracellular electron transfer associated with denitrification was identified as the primary limiting step with a formal potential of -0.225 ± 0.007 V vs. Ag/AgCl sat. KCl at pH 7 and 25 °C. By varying the nitrate concentration, it was revealed that the biocathode exhibits a strong affinity for nitrate (KMapp of 0.7 ± 0.2 mg N-NO3- L-1). The maximum denitrification rate was observed at a pH of 6 and a temperature of 35 °C. Furthermore, the findings highlight a 2e-/1H+ transfer, which holds considerable implications for the energy metabolism of bioelectrochemical denitrifiers. These compiled results provide valuable insights into the understanding of denitrifying biocathodes and enable the improvement and prediction of their performance.

Electroactive microorganisms are now understood to be abundant across nature, though many are categorized as \"weak electricigens\" not suitable for reasonable power generation. We report the use of weak electricigens from the natural environment for rapid, real-time water quality monitoring. Using a variety of pesticides as model chemical pollutants, the bioelectrochemical sensor was responsive within minutes at all concentrations tested (0.05-2 ppm) and could be repreatedly used long-term. Due to the prevalence of electroactive microorganisms in the natural environment, such sensors could work in tandem with conventional monitoring methods and may be useful for detecting emerging contaminants.

Bidirectional extracellular electron transfer (EET) is crucial to upholding microbial metabolism with insoluble electron acceptors or donors in anoxic environments. Investigating bidirectional EET-capable microorganisms is desired to understand the cell-cell and microbe-mineral interactions and their role in mineral cycling besides leveraging their energy generation and conversion, biosensing, and bio-battery applications. Here, we report on iron cycling by haloalkaliphilic Geoalkalibacter halelectricus via bidirectional EET under haloalkaline conditions. It efficiently reduces Fe3+ oxide (Fe2O3) to Fe0 at a 0.75 ± 0.08 mM/mgprotein/d rate linked to acetate oxidation via outward EET and oxidizes Fe0 to Fe3+ at a 0.24 ± 0.03 mM/mgprotein/d rate via inward EET to reduce fumarate. Bioelectrochemical cultivation confirmed its outward and inward EET capabilities. It produced 895 ± 23 µA/cm2 current by linking acetate oxidation to anode reduction via outward EET and reduced fumarate by drawing electrons from the cathode (‒2.5 ± 0.3 µA/cm2) via inward EET. The cyclic voltammograms of G. halelectricus biofilms revealed redox moieties with different formal potentials, suggesting the involvement of different membrane components in bidirectional EET. The cyclic voltammetry and GC-MS analysis of the cell-free spent medium revealed the lack of soluble redox mediators, suggesting direct electron transfer by G. halelecctricus in achieving bidirectional EET. By reporting on the first haloalkaliphilic bacterium capable of oxidizing and reducing insoluble Fe0 and Fe3+ oxide, respectively, this study advances the limited understanding of the metabolic capabilities of extremophiles to respire on insoluble electron acceptors or donors via bidirectional EET and invokes the possible role of G. halelectricus in iron cycling in barely studied haloalkaline environments. IMPORTANCE Bidirectional extracellular electron transfer (EET) appears to be a key microbial metabolic process in anoxic environments that are depleted in soluble electron donor and acceptor molecules. Though it is an ecologically important and applied microbial phenomenon, it has been reported with a few microorganisms, mostly from nonextreme environments. Moreover, direct electron transfer-based bidirectional EET is studied for very few microorganisms with electrodes in engineered systems and barely with the natural insoluble electron acceptor and donor molecules in anoxic conditions. This study advances the understanding of extremophilic microbial taxa capable of bidirectional EET and its role in barely investigated Fe cycling in highly saline-alkaline environments. It also offers research opportunities for understanding the membrane components involved in the bidirectional EET of G. halelectricus. The high rate of Fe3+ oxide reduction activity by G. halelectricus suggests its possible use as a biocatalyst in the anaerobic iron bioleaching process under neutral-alkaline pH conditions.

Research on electroactive microorganisms (EAM) often focuses either on their physiology and the underlying mechanisms of extracellular electron transfer or on their application in microbial electrochemical technologies (MET). Thermodynamic understanding of energy conversions related to growth and activity of EAM has received only a little attention. In this study, we aimed to prove the hypothesized restricted energy harvest of EAM by determining biomass yields by monitoring growth of acetate-fed biofilms presumably enriched in Geobacter, using optical coherence tomography, at three anode potentials and four acetate concentrations. Experiments were concurrently simulated using a refined thermodynamic model for EAM. Neither clear correlations were observed between biomass yield and anode potential nor acetate concentration, albeit the statistical significances are limited, mainly due to the observed experimental variances. The experimental biomass yield based on acetate consumption (YX/ac = 37 ± 9 mgCODbiomass gCODac-1) was higher than estimated by modeling, indicating limitations of existing growth models to predict yields of EAM. In contrast, the modeled biomass yield based on catabolic energy harvest was higher than the biomass yield from experimental data (YX/cat = 25.9 ± 6.8 mgCODbiomass kJ-1), supporting restricted energy harvest of EAM and indicating a role of not considered energy sinks. This calls for an adjusted growth model for EAM, including, e.g., the microbial electrochemical Peltier heat to improve the understanding and modeling of their energy metabolism. Furthermore, the reported biomass yields are important parameters to design strategies for influencing the interactions between EAM and other microorganisms and allowing more realistic feasibility assessments of MET.

The poor bacterial loading capacity and biocompatibility of the anode lead to weak electricity production performance of microbial fuel cells (MFCs). Inspired by kelp, we developed a double-layer hydrogel bioanode based on sodium alginate (SA). The inner hydrogel layer of encapsulated Fe3O4 and electroactive microorganisms (EAMs) was used as the bioelectrochemical catalytic layer. The outer hydrogel layer formed by cross-linking SA with polyvinyl alcohol (PVA) was used as the protective layer. The 3D porous structure of the inner hydrogel formed based on Fe3O4 facilitated the electroactive bacteria colonization and electron transfer, while the high structural toughness, salt-resistance and antibacterial properties of the outer highly cross-linked hydrogel served to protect the catalytic layer for stable electricity production. When high-salt waste leachate was used as the nutrient, the amazing open-circuit voltage (OCV) of 1.17 V and the operating voltage of 781 mV were brought by the double-layer hydrogel bioanode PVA@SA&Fe3O4/EAMs@SA.