Sorting Out the Latest Advances in Separators and Pilot-Scale Microbial Electrochemical Systems for Wastewater Treatment: Concomitant Development, Practical Application, and Future Perspective.

To date, dozens of pilot-scale microbial fuel cell (MFC) devices have been successfully developed worldwide for treating various types of wastewater. The availability and configurations of separators are determining factors for the economic feasibility, efficiency, sustainability, and operability of these devices. Thus, the concomitant advances between the separators and pilot-scale MFC configurations deserve further clarification. The analysis of separator configurations has shown that their evolution proceeds as follows: from ion-selective to ion-non-selective, from nonpermeable to permeable, and from abiotic to biotic. Meanwhile, their cost is decreasing and their availability is increasing. Notably, the novel MFCs configured with biotic separators are superior to those configured with abiotic separators in terms of wastewater treatment efficiency and capital cost. Herein, a highly comprehensive review of pilot-scale MFCs (>100 L) has been conducted, and we conclude that the intensive stack of the liquid cathode configuration is more advantageous when wastewater treatment is the highest priority. The use of permeable biotic separators ensures hydrodynamic continuity within the MFCs and simplifies reactor configuration and operation. In addition, a systemic comparison is conducted between pilot-scale MFC devices and conventional decentralized wastewater treatment processes. MFCs showed comparable cost, higher efficiency, long-term stability, and significant superiority in carbon emission reduction. The development of separators has greatly contributed to the availability and usability of MFCs, which will play an important role in various wastewater treatment scenarios in the future.

Using stacked pot connection of wetland microbial fuel cells to charge the battery: Potential and effecting factor.

In the practical application of wetland microbial fuel cells (WMFCs), suitable designs and stacked connection systems have consistently been employed to increase and harvest power generation. Our study compares different WMFCs designs and demonstrates that the cylinder pot design outperforms the small hanging pot design in terms of electrical energy production. Moreover, power generation from the cylinder pot can be further optimized through separator modification and stacked connections. The stacked WMFCs design exhibited no voltage reversal, with an average power output ranging from 0.03 ± 0.01 mW (single pot) to 0.11 ± 0.05 mW (stacked connection of 5 pots) over a 60-day operational period. Additionally, our study identifies distinct patterns in both anodic and cathodic physiochemical factors including electrical conductivity (EC), pH, and nitrate (NO3-), highlighting the significant influence of plant involvement on altering concentrations and levels in different electrode zones. The WMFCs bioelectricity production system, employing 15 pots stacked connections achieves an impressive maximum power density of 9.02 mW/m2. The system's practical application is evidenced by its ability to successfully power a DC-DC circuit and charge a 1.2 V AAA battery over a period of 30 h, achieving an average charging rate of 0.0.2 V per hour.

Prevalence of Escherichia coli in electrogenic biofilm on activated carbon in microbial fuel cell.

For a better understanding of the distribution of depth-dependent electrochemically active bacteria at in the anode zone, a customized system in a microbial fuel cell (MFC) packed with granular activated carbon (GAC) was developed and subsequently optimized via electrochemical tests. The constructed MFC system was sequentially operated using two types of matrice solutions: artificially controlled compositions (i.e., artificial wastewater, AW) and solutions obtained directly from actual sewage-treating municipal plants (i.e., municipal wastewater, MW). Notably, significant difference(s) of system efficiencies between AW or MW matrices were observed via performance tests, in that the electricity production capacity under MW matrices is < 25% that of the AW matrices. Interestingly, species of Escherichia coli (E. coli) sampled from the GAC bed (P1: deeper region in GAC bed, P2: shallow region of GAC near electrolytes) exhibited an average relative abundance of 75 to 90% in AW and a relative abundance of approximately 10% in MW, while a lower relative abundance of E. coli was found in both the AW and MW anolyte samples (L). Moreover, similar bacterial communities were identified in samples P1 and P2 for both the AW and MW solutions, indicating a comparable distribution of bacterial communities over the anode area. These results provide new insights into E. coli contribution in power production for the GAC-packed MFC systems (i.e., despite the low contents of Geobacter (> 8%) and Shewanella (> 1%)) for future applications in sustainable energy research. KEY POINTS: • A microbial community analysis for depth-dependence in biofilm was developed. • The system was operated with two matrices; electrochemical performance was assessed. • E. coli spp. was distinctly found in anode zone layers composed of activated carbon.

Enhancing diversified extracellular electron transfer (EET) processes through N-MXene-modified non-adhesive hydrogel bioanodes.

The focus of this study is to develop a high-performance anode material for microbial fuel cells (MFCs). PEDOT:PSS and nitrogen-modified MXene were combined to create a hydrogel composite material called PPNM, which was drop-cast onto carbon felt (CF) as the MFCs anode. The PPNM exhibited a higher peak power density of 4.78 W m-2, an increase of 332% compared to the CF anode. It is worth noting that the PPNM Hydrogel maintains its rough and porous structure, providing favorable sites for bacterial colonization. The introduction of N-MXene has improved the electrochemical performance of the hydrogel, particularly impacting the mediated electron transfer process. Microbial community analysis revealed the presence of more electrochemically active species on the PPNM anode. These findings highlight the potential of PPNM hydrogel and pave the way for similar strategies in achieving high-performance anodes in MFCs.

Efficient degradation of carbamazepine and metagenomic investigations of anodic biofilm in microbial fuel cells.

Environmental contamination with carbamazepine is a considerable global problem. In this study, two-compartment microbial fuel cells (MFCs) were constructed to investigate the degradation performance of carbamazepine, and the degradation mechanism was further explored by using metagenomic analysis. The results showed that MFCs exhibited excellent carbamazepine removal performance and also generated electricity. The removal rate of carbamazepine reached 73.56% over the 72-h operation period, which was 3.09 times higher than that of the traditional anaerobic method, and the peak voltage of the MFCs could reach 416 mV. Metagenomics revealed significant differences in microbial community composition between MFCs and the traditional anaerobic method (p < 0.05), and Proteobacteria (81.57%) was predominant bacterial phyla during the degradation of carbamazepine by MFCs. Among them, the microbial communities at the genus level were mainly composed of Pseudomonas, Pusillimonas, Burkholderia, Stenotrophomonas, Methyloversatilis and Nitrospirillum. Kyoto Encyclopedia of genes and genomes (KEGG) metabolic pathway analysis showed that the number of genes related to carbon and nitrogen metabolism increased by 85.12% and 142.25%, respectively. Importantly, a greater number of genes of microbial grown on the surface of anode were assigned to denitrification and the degradation of aromatic compounds. This research provides a cost-effective method for treating wastewater contaminated with carbamazepin.

Long-term evaluation of soil-based bioelectrochemical green roof systems for greywater treatment.

Water scarcity has generated the need to identify new sources. Due to its low organic contaminant load, greywater reuse has emerged as a potential alternative. Moreover, the search for decentralized treatment systems in urban areas has prompted research on using green roofs for greywater treatment. However, the performance of organic matter removal is limited by the type of substrate and height of the growing media. Bioelectrochemical systems (BESs) improve treatment performance by providing an additional electron acceptor (the electrode). In this study, nine reactors under three different conditions, i.e., open circuit (OC), microbial fuel cell (MFC), and microbial electrolysis cell (MEC), were built to evaluate the treatment of synthetic greywater in a substrate-growing medium composed of perlite and coconut fiber and operated in batch-cycle mode for 397 days. The results suggested that using BESs enables greywater treatment and the removal of pollutants to levels that allow their reuse for irrigation. Furthermore, electrical conductivity was reduced from 732.4 ± 41.2 µS/cm2 in OC to 637.32 ± 22.73 µS/cm2 and 543.15 ± 19.69 µS/cm2 in MEC and MFC, respectively. The soluble chemical oxygen demand in the latter treatments reached 76% removal, compared to levels above the OC, which only reached approximately 67%. Microbial community analysis revealed differences, mainly in the cathodes, showing a higher development of Flavobacterium, Azospirillum, and Zoogloea in MFCs, which could explain the higher levels of organic matter removal in the other conditions, suggesting that the BES could produce an enrichment of beneficial bacterial groups for treatment. Therefore, implementing BESs in green roofs enables sustainable long-term greywater treatment.

Biohydrogen upgradation and wastewater treatment in 3-chambered bioelectrochemical system assisted with H2/O2-based redox reactions.

The current need for the upgradation of biohydrogen generation and contaminant removal in two-chambered microbial electrolysis cells (MECs) compels the design of alternatives i.e. bioelectrochemical systems (BESs) to conventional reactors. In this study, a novel three-chambered design of MEC (BES-1) was developed with a common anodic chamber and a two-cathodic chambers at both ends of the anodic chamber, separated by a membrane (MEC-MEC). To facilitate electricity recovery, a microbial fuel cell (MFC) was integrated with an MEC in BES-2. Cathodic hydrogen recovery of 8.89 and 4.81 mL/L.day, and organic matter removal of 82% and 76% were obtained in BES-1 and BES-2, respectively, demonstrating their capabilities for bioremediation. Electrochemical analyses also revealed that cathodic reduction reactions improved with the effective utilization of protons during integration. Our design regulates H2/O2-associated electrochemical reactions and is beneficial for maintaining pH equilibrium. From cost and energy perspectives, the integrated BES provides a platform for two different reactions simultaneously and is capable of boosting overall hydrogen recovery and organic matter removal. Moreover, the compactness and competitiveness of such an integrated BES increase its scope for real-world applications.

Revealing the performance of aerotolerant anodes for electroactive nitrification/denitrification and current production under coexistence of oxygen and nitrate conditions.

The coexistence of oxygen and/or nitrate at anode usually affects the biofilm activities of traditional anaerobic anode, thereby deteriorating wastewater treatment performance of microbial fuel cells (MFCs). Improving the aerotolerant responses of anode biofilms is a challenge for field application. In this study, we report that using the electroactive nitrifying/denitrifying inoculum and air-cathode expansion could fabricate the aerotolerant anode biofilms (AAB) under affordable nitrate stress (90 ± 5 mg/L). The highest average removal efficiencies were 99% for chemical oxygen demand (COD), NH4+-N and total nitrogen. The highest average current output of 0.69 mA and power density of 290 mW/m2 were obtained. The average current was confirmed to be reduced 10%-78% but the power density remained almost stable except the quart-air-cathodes MFC by increasing dissolved oxygen concentration with expansion of the air-cathode area. The higher oxygen concentration also contributed to oxidation of ammonium through electroactive autotrophic nitrification. The facultative anaerobic bacteria including Thauera, Microsillaceae, Shinella, Blastocatellaceae, Rhodobacter, Comamonadaceae, Caldilineaceae were enriched, which forms the AAB to remove nitrogen and produce current. Therefore, an easy-to-use method to fabricate AAB is evaluated to realize practical applications of MFCs in wastewater treatment.

Reduction of Toxic Metal Ions and Production of Bioelectricity through Microbial Fuel Cells Using Bacillus marisflavi as a Biocatalyst.

Industrialization has brought many environmental problems since its expansion, including heavy metal contamination in water used for agricultural irrigation. This research uses microbial fuel cell technology to generate bioelectricity and remove arsenic, copper, and iron, using contaminated agricultural water as a substrate and Bacillus marisflavi as a biocatalyst. The results obtained for electrical potential and current were 0.798 V and 3.519 mA, respectively, on the sixth day of operation and the pH value was 6.54 with an EC equal to 198.72 mS/cm, with a removal of 99.08, 56.08, and 91.39% of the concentrations of As, Cu, and Fe, respectively, obtained in 72 h. Likewise, total nitrogen concentrations, organic carbon, loss on ignition, dissolved organic carbon, and chemical oxygen demand were reduced by 69.047, 86.922, 85.378, 88.458, and 90.771%, respectively. At the same time, the PDMAX shown was 376.20 ± 15.478 mW/cm2, with a calculated internal resistance of 42.550 ± 12.353 Ω. This technique presents an essential advance in overcoming existing technical barriers because the engineered microbial fuel cells are accessible and scalable. It will generate important value by naturally reducing toxic metals and electrical energy, producing electric currents in a sustainable and affordable way.

High-Performance Macroporous Free-Standing Microbial Fuel Cell Anode Derived from Grape for Efficient Power Generation and Brewery Wastewater Treatment.

Microbial fuel cells (MFCs) have the potential to directly convert the chemical energy in organic matter into electrical energy, making them a promising technology for achieving sustainable energy production alongside wastewater treatment. However, the low extracellular electron transfer (EET) rates and limited bacteria loading capacity of MFCs anode materials present challenges in achieving high power output. In this study, three-dimensionally heteroatom-doped carbonized grape (CG) monoliths with a macroporous structure were successfully fabricated using a facile and low-cost route and employed as independent anodes in MFCs for treating brewery wastewater. The CG obtained at 900 °C (CG-900) exhibited excellent biocompatibility. When integrated into MFCs, these units initiated electricity generation a mere 1.8 days after inoculation and swiftly reached a peak output voltage of 658 mV, demonstrating an exceptional areal power density of 3.71 W m-2. The porous structure of the CG-900 anode facilitated efficient ion transport and microbial community succession, ensuring sustained operational excellence. Remarkably, even when nutrition was interrupted for 30 days, the voltage swiftly returned to its original level. Moreover, the CG-900 anode exhibited a superior capacity for accommodating electricigens, boasting a notably higher abundance of Geobacter spp. (87.1%) compared to carbon cloth (CC, 63.0%). Most notably, when treating brewery wastewater, the CG-900 anode achieved a maximum power density of 3.52 W m-2, accompanied by remarkable treatment efficiency, with a COD removal rate of 85.5%. This study provides a facile and low-cost synthesis technique for fabricating high-performance MFC anodes for use in microbial energy harvesting.

Fabrication of a Molybdenum Dioxide/Multi-Walled Carbon Nanotubes Nanocomposite as an Anodic Modification Material for High-Performance Microbial Fuel Cells.

A nanocomposite of multi-walled carbon nanotubes (MWCNTs) decorated with molybdenum dioxide (MoO2) nanoparticles is fabricated through the reduction of phosphomolybdic acid hydrate on functionalized MWCNTs in a hydrogen-argon (10%) atmosphere in a tube furnace. The MoO2/MWCNTs composite is proposed as an anodic modification material for microbial fuel cells (MFCs). MWCNTs have outstanding physical and chemical peculiarities, with functionalized MWCNTs having substantially large electroactive areas. In addition, combined with the exceptional properties of MoO2 nanoparticles, the synergistic advantages of functionalized MWCNTs and MoO2 nanoparticles give a MoO2/MWCNTs anode a large electroactive area, excellent electronic conductivity, enhanced extracellular electron transfer capacity, and improved nutrient transfer capability. Finally, the power harvesting of an MFC with the MoO2/MWCNTs anode is improved, with the MFC showing long-term repeatability of voltage and current density outputs. This exploratory research advances the fundamental application of anodic modification to MFCs, simultaneously providing valuable guidance for the use of carbon-based transition metal oxide nanomaterials in high-performance MFCs.

Migration of various ions based on pH shifts triggered by the application of sediment microbial fuel cells.

Sediment microbial fuel cells (SMFCs) represent a technology that can enhance sediment quality through processes such as nutrient suppression while simultaneously generating electricity from microorganisms. Despite its importance in elucidating the principles of nutrient suppression, the complex behavior of various ions within this context has been rarely explored. Herein, we applied an SMFC and systematically evaluated alterations in ion concentrations in interstitial and overlying waters. The SMFC deployment substantially decreased Na+ concentrations and increased Cl- levels in the interstitial water. This intriguing phenomenon was attributed to reactions driven by the electrodes. These reactions induced remarkable shifts in pH. Consequently, this pH shift triggered the leaching of heavy metals, particularly Fe, and decreased HCO3- concentrations within the interstitial water, thereby inducing the migration of other ions, including Na+ and Cl-, as compensation. Moreover, the PO43- concentration in interstitial water showed an increasing trend upon SMFC application, which contradicts the results of several previous reports. This increase was primarily attributed to the release of PO43-caused by the leaching of Fe salts, which was triggered by the pH shift. These findings provide new insights into sediment improvement research through SMFCs, enhancing our understanding of the fundamental principles and broadening the potential applications of this technology.

Current advances of chlorinated organics degradation by bioelectrochemical systems: a review.

Chlorinated organic compounds (COCs) are typical refractory organic compounds, having high biological toxicity. These compounds are a type of pervasive pollutants that can be present in polluted soil, air, and various types of waterways, such as groundwater, rivers, and lakes, posing a significant threat to the ecological environment and human health. Bioelectrochemical systems (BESs) are an effective strategy for the degradation of bio-refractory compounds. BESs improve the waste treatment efficiency through the application of weak electrical stimulation. This review discusses the processes of BESs configurations and degradation performances in different environmental media including wastewater, soil, waste gas and groundwater. In addition, the degradation mechanisms and performance-enhancing additives are summarized. The future challenges and perspectives on the development of BES for COCs removal are briefly discussed.

Progress in enhancing the remediation performance of microbial fuel cells for contaminated groundwater.

Microbial fuel cells (MFCs) have become more prevalent in groundwater remediation due to their capacity for power generation, removal of pollution, ease of assembly, and low secondary contamination. It is currently being evaluated for practical application in an effort to eliminate groundwater pollution. However, a considerable majority of research was conducted in laboratories. But the operational circumstances including anaerobic characteristics, pH, and temperature vary at different sites. In addition, the complexity of contaminants and the positioning of MFCs significantly affect remediation performance. Taking the aforementioned factors into consideration, this review summarizes a bibliography on the application of MFCs for the remediation of groundwater contamination during the last ten decades and assesses the impact of environmental conditions on the treatment performance. The design of the reactor, including configuration, dimensions, electrodes, membranes, separators, and target contaminants are discussed. This review aims to provide practical guidance for the future application of MFCs in groundwater remediation.

Moisture-Enabled Germination of Heat-Activated Bacillus Endospores for Rapid and Practical Bioelectricity Generation: Toward Portable, Storable Bacteria-Powered Biobatteries.

Small-scale battery-like microbial fuel cells (MFCs) are a promising alternative power source for future low-power electronics. Controllable microbial electrocatalytic activity in a miniaturized MFC with unlimited biodegradable energy resources would enable simple power generation in various environmental settings. However, the short shelf-life of living biocatalysts, few ways to activate the stored biocatalysts, and extremely low electrocatalytic capabilities render the miniature MFCs unsuitable for practical use. Here, heat-activated Bacillus subtilis spores are revolutionarily used as a dormant biocatalyst that can survive storage and rapidly germinate when exposed to special nutrients that are preloaded in the device. A microporous, graphene hydrogel allows the adsorption of moisture from the air, moves the nutrients to the spores, and triggers their germination for power generation. In particular, forming a CuO-hydrogel anode and an Ag2 O-hydrogel cathode promotes superior electrocatalytic activities leading to an exceptionally high electrical performance in the MFC. The battery-type MFC device is readily activated by moisture harvesting, producing a maximum power density of 0.4 mW cm-2 and a maximum current density of 2.2 mA cm-2 . The MFC configuration is readily stackable in series and a three-MFC pack produces enough power for several low-power applications, demonstrating its practical feasibility as a sole power source.

Investigating Variability in Microbial Fuel Cells.

In scientific studies, replicas should replicate, and identical conditions should produce very similar results which enable parameters to be tested. However, in microbial experiments which use real world mixed inocula to generate a new "adapted" community, this replication is very hard to achieve. The diversity within real-world microbial systems is huge, and when a subsample of this diversity is placed into a reactor vessel or onto a surface to create a biofilm, stochastic processes occur, meaning there is heterogeneity within these new communities. The smaller the subsample, the greater this heterogeneity is likely to be. Microbial fuel cells are typically operated at a very small laboratory scale and rely on specific communities which must include electrogenic bacteria, known to be of low abundance in most natural inocula. Microbial fuel cells (MFCs) offer a unique opportunity to investigate and quantify variability as they produce current when they metabolize, which can be measured in real time as the community develops. In this research, we built and tested 28 replica MFCs and ran them under identical conditions. The results showed high variability in terms of the rate and amount of current production. This variability perpetuated into subsequent feeding rounds, both with and without the presence of new inoculate. In an attempt to control this variability, reactors were reseeded using established "good" and "bad" reactors. However, this did not result in replica biofilms, suggesting there is a spatial as well as a compositional control over biofilm formation. IMPORTANCE The research presented, although carried out in the area of microbial fuel cells, reaches an important and broadly impacting conclusion that when using mixed inoculate in replica reactors under replicated conditions, different communities emerge capable of different levels of metabolism. To date there has been very little research focusing on this, or even reporting it, with most studies using duplicate or triplicate reactors, in which this phenomenon is not fully observed. Publishing data in which replicas do not replicate will be an important and brave first step in the research into understanding this fundamental microbial process.

Functional Nanomaterial-Modified Anodes in Microbial Fuel Cells: Advances and Perspectives.

Microbial fuel cell (MFC) is a promising approach that could utilize microorganisms to oxidize biodegradable pollutants in wastewater and generate electrical power simultaneously. Introducing advanced anode nanomaterials is generally considered as an effective way to enhance MFC performance by increasing bacterial adhesion and facilitating extracellular electron transfer (EET). This review focuses on the key advances of recent anode modification materials, as well as the current understanding of the microbial EET process occurring at the bacteria-electrode interface. Based on the difference in combination mode of the exoelectrogens and nanomaterials, anode surface modification, hybrid biofilm construction and single-bacterial surface modification strategies are elucidated exhaustively. The inherent mechanisms may help to break through the performance output bottleneck of MFCs by rational design of EET-related nanomaterials, and lead to the widespread application of microbial electrochemical systems.

Analysis and enhancement of the energy utilization efficiency of corn stover using strain Lsc-8 in a bioelectrochemical system.

The strain Lsc-8 can produce a current density of 33.08 µA cm-2 using carboxymethylcellulose (CMC) as a carbon source in a three-electrode configuration. A co-culture system of strain Lsc-8 and Geobacter sulfurreducens PCA was used to efficiently convert cellulose into electricity to improve the electricity generation capability of microbial fuel cells (MFCs). The maximum current density achieved by the co-culture with CMC was 559 µA cm-2, which was much higher than that of strain Lsc-8 using CMC as the carbon source. The maximum power density reached 492.05 ± 52.63 mW cm-2, which is much higher than that previously reported. Interaction mechanism studies showed that strain Lsc-8 had the ability to secrete riboflavin and convert cellulose into acetic acid, which might be the reason for the high electrical production performance of the co-culture system. In addition, to the best of our knowledge, a co-culture or single bacteria system using agricultural straw as the carbon source to generate electricity has not been reported. In this study, the maximum current density of the three-electrode system inoculated with strain Lsc-8 was 14.56 µA cm-2 with raw corn stover as the sole carbon source. Raw corn stover as a carbon source was also investigated for use in a co-culture system. The maximum current density achieved by the co-culture was 592 µA cm-2. The co-culture system showed a similar electricity generation capability when using raw corn stover and when using CMC. This research shows for the first time that a co-culture or single bacteria system can realize both waste biomass treatment and waste power generation.

Novel slow-release carbon source improves anodic denitrification and electricity generation efficiency in microbial fuel cells.

MFC anodic denitrification is more suitable for the coexistence of organic matter and nitrate in actual sewage, but the traditional carbon source has some problems such as high cost and difficulty of dosage control in MFC. Herein, corncob and polycaprolactone (PCL) were mechanically pulverized and mixed in the system of polyvinyl alcohol and sodium alginate, and cross-linked to prepare slow-release carbon source fillers (CPSP), which were added to the MFC anolyte to realize the coupling of solid-phase denitrification and anodic denitrification. Results showed the start-up period of MFC experimental group (MFC-C) with CPSP was slightly longer than the control group (MFC-0), but MFC-C's maximum output voltage (648.4 mV) and power density (2738 mW/m3) could be increased by 5% and 15% higher than that of MFC-0 (P < 0.05). The degradation process of MFC substrate in unit cycle was mainly divided into nitrogen removal stage (0-8 h) and electricity generation stage (8-48 h). The NO3--N and COD degradation and power generation kinetic processes of MFC conformed to the Han-Levenspiel model. Kinetics experiments showed CPSP can improve the affinity and tolerance of MFC to NO3--N, also it can alleviate the pressure of electron competition in anolyte and improve coulombic efficiency. In addition, microbial communities were significantly changed under the effect of CPSP (P < 0.001). Meanwhile, CPSP can promote the synthesis of denitrification functional genes. This study provides a new strategy to improve the performance of MFC by the addition of novel denitrification carbon source.

Iron/iron carbide coupled with S, N co-doped porous carbon as effective oxygen reduction reaction catalyst for microbial fuel cells.

As a novel energy device, microbial fuel cells (MFCs) have attracted much attention for their dual functions of electricity generation and sewage treatment. However, the sluggish oxygen reduction reaction (ORR) kinetic on the cathode have hindered the practical application of MFCs. In this work, metallic organic framework derived carbon framework co-doped by Fe, S, N tri-elements was used as alternative electrocatalyst to the conventional Pt/C cathode catalyst in pH-universal electrolytes. The amount of thiosemicarbazide from 0.3 to 3 g determined the surface chemical property, and therefore the ORR activity of FeSNC catalysts. The sulfur/nitrogen doping and Fe/Fe3C embedded in carbon shell was characterized by X-ray photoelectron spectroscopy and transmission electron microscopy. The synergy of iron salt and thiosemicarbazide contributed to the improvement of nitrogen and sulfur doping. Sulfur atoms were successfully doped into the carbon matrix and formed a certain amount of thiophene- and oxidized-sulfur. The optimal FeSNC-3 catalyst synthesized with 1.5 g of thiosemicarbazide exhibited the highest ORR activity with a positive half wave potential of 0.866 V in alkaline and 0.691 V (vs. Reversible Hydrogen Electrode) in neutral electrolyte, which both outperformed the commercial Pt/C catalyst. However, as the amount of thiosemicarbazide surpassed 1.5 g, the catalytic performance of FeSNC-4 was lowered, and this could be assigned to the decreased defects and low specific surface area. The excellent ORR performance in neutral medium urged FeSNC-3 as good cathode catalyst in single chambered MFC (SCMFC). It showed the highest maximum power density of 2126 ± 100 mW m-2, excellent output stability of 8.14% decline in 550 h, chemical oxygen demand removal of 90.7 ± 1.6% and coulombic efficiency of 12.5 ± 1.1%, all superior to those of benchmark SCMFC-Pt/C (1637 ± 35 mW m-2, 15.4%, 88.9 ± 0.9%, and 10.2 ± 1.1%). These outstanding results were associated to the large specific surface area and synergistic interaction of multiple active sites, like Fe/Fe3C, Fe-N4, pyridinic N, graphite N and thiophene-S.