Bioresource technology • 2025

The growing demand for sustainable wastewater valorization and renewable hydrogen energy has amplified the need for systems capable of recovering energy and producing hydrogen without external electricity input. In this study, ten single microbial fuel cells (MFC) were connected in parallel to enhance power generation, and the electricity produced was controlled using a power management system. The energy was then directly supplied to a microbial electrolysis cell (MEC) in real time and stored in a battery prior to the MEC operation. When directly connected, the MFC-MEC system achieved a hydrogen production rate of 0.08 m 3 /m 3 /d, while the battery-assisted mode reached 0.66 m 3 /m 3 /d, demonstrating an eight-fold increase. These findings demonstrate the feasibility of integrated MFC-MEC systems for self-powered hydrogen production, offering a potential pathway for transforming organic wastewater into hydrogen as a renewable energy carrier.

Bioelectrochemistry (Amsterdam, Netherlands) • 2025

Electroanalytical techniques can be used to provide new insights into the electrochemical response of microbial bioelectrodes with some limitations due to complex interphase biofilm-electrode. In this work, characterization techniques, including open-circuit voltage, cyclic voltammetry, and electrochemical impedance spectroscopy, were applied to the bioanode, while chronoamperometry was used for the MEC with nine combinations of applied voltage and biomass content for anodic biogas production. The open-circuit potential was characteristic of the bioanode at -0.4 V ± 0.02 V vs. Ag/AgCl; the cumulative charge evolution allowed differentiation of operation conditions, reaching a total charge between 30C and 170C. Three distinct profiles were found among the voltammograms including oxidation peaks, oxidation waves and no faradaic signals. The charge transfer resistance estimated via electrochemical impedance spectroscopy only allowed differentiation of the final from the initial conditions. A nonlinear correlation was found between electrochemical parameters and biogas volume and its composition. This work presents a method for the systematic electroanalysis of bioelectrodes and provides information on their behavior under different biomass and applied voltage conditions.

Food research international (Ottawa, Ont.) • 2025

S. aureus is a common foodborne pathogen that poses a great danger to the food industry and human health. Slightly acidic electrolytic water (SAEW) is characterized by strong antimicrobial effect and high potential for application. In this study, the effect of SAEW on the formation of VBNC state of S. aureus was systematically evaluated, and the differences in the formation of VBNC state of S. aureus in planktonic and biofilm states were also analyzed by flow cytometry. The results showed that treatment of S. aureus in planktonic and biofilm states with 0.8 mg/L SAEW resulted in complete loss of culturability at 2.5 h and 2.0 h, respectively, with 3.56 ± 0.28 % and 4.81 ± 0.13 % of cells formed VBNC state. And both states of VBNC S. aureus could be resuscitated, and the planktonic VBNC state S. aureus resuscitated faster. ATP concentration, ROS level and respiration intensity of different states of bacteria were examined, compared with uninduced bacteria, VBNC S. aureus showed increased ATP concentration and ROS level, and decreased catalase activity, esterase activity, and respiratory intensity. Meanwhile, the overall metabolic level of the biofilm VBNC state bacteria was lower than that of the planktonic VBNC state bacteria, and its ROS level was higher than that of the planktonic VBNC state bacteria. Gene expression was determined by RT-qPCR, and the results showed that the transcript levels of genes regulating N-acetylglucosamine metabolism were up-regulated and those related to lipid metabolism, antioxidant and respiratory metabolism were down-regulated in the VBNC state S. aureus. This study comprehensively evaluates the application effect of SAEW in the food industry to provide a scientific basis for its safe and effective use; deepens the understanding of the mechanism of the formation of VBNC states by bacteria in different states, and provides theoretical support for the control of VBNC bacteria.

Bioelectrochemistry (Amsterdam, Netherlands) • 2025

Microbial electrolysis cells (MECs) are promising for biohydrogen production from waste streams, yet their overall energy efficiency requires enhancement to be viable. This study, for the first time, utilized double-chamber MECs with graphite (Gr), ruthenium-coated graphite (Ru), and palladium-coated graphite (Pd) cathodes for biohydrogen production from potato processing wastewater under various applied voltages. The highest hydrogen evolution rate (HER) and energy efficiency (ηₑ) were achieved in the Ru reactor at 1.0 V, reaching 0.43 LH₂ L -1 d -1 and 106 %, respectively. These represent improvements of 81 % in HER and 33 % in ηₑ compared to the uncoated Gr reactor. The surface morphology of the Ru and Pd coated electrodes was characterized by SEM and XRD. Next-generation sequencing of the anode biofilm indicated a microbial community comprising polysaccharide-consuming bacteria, such as the phylum Bacteroidetes, and key exoelectrogens, including the genus Geobacter. An Artificial Neural Network (ANN) model (R 2  = 0.9432) was also developed to simulate HER performance, confirming high performance of Ru-coated cathode under various voltages. A techno-economic analysis (TEA) was conducted to assess the effect of cathode coating on the payback period. The analysis revealed that Pd and Ru coatings reduced the payback period by approximately 36 % and 34 %, respectively.

Bioelectrochemistry (Amsterdam, Netherlands) • 2025

This study delves deeply into the impact of different electrode materials on the redox properties of extracellular polymeric substances (EPS) within electroactive mixed microbial communities. The experimental results reveal that the redox properties of EPS exhibit significant variations depending on the electrode materials employed. Among them, the reactor (MEC-BTF3) utilizing dual carbon felts as the cathode and anode electrodes demonstrates the most outstanding degradation performance. Within a 96-h reaction period, the degradation efficiency of trichloroethylene (TCE) by this reactor reaches as high as 94.3 %. In-depth analysis indicates that the dual carbon electrodes possess a low charge transfer internal resistance, a characteristic that substantially enhances the electron transfer rate. Moreover, in the protein secondary structure of EPS, the β-sheet structure accounts for 40.0 % of the total amide I region. This structural feature effectively promotes the formation and stability of microbial aggregates. The important findings of this study not only facilitate a more profound understanding of the interaction mechanisms between electrode materials and EPS in electroactive microbial communities but also offer theoretical guidance for the development of more efficient and targeted bioelectrochemical system (BES).

Applied biochemistry and biotechnology • 2025

Redox mediators (RMs) have been widely employed in bioelectrochemical systems to enhance electron transfer efficiency. However, systematic comparisons of RM-driven microbial selectivity and its direct correlation with methane production in microbial electrolysis cells (MECs) remain unexplored. This study investigates methane production in MECs using carbon felt (CF) electrodes modified with four RMs, i.e., neutral red (NR), anthraquinone-2,6-disulfonic acid disodium salt (AQDS), humic acid (HA), methyl viologen (MV), and the conductive polymer polyaniline (PANI). Cyclic voltammetry and electrochemical impedance spectroscopy revealed superior electrochemical activity for NR- and HA-modified electrodes (CF-NR, CF-HA) among the tests. CF-NR and CF-PANI demonstrated the highest biocompatibility, supporting 25% and 15% greater biofilm biomass than unmodified CF, respectively. Modified electrodes exhibited lower alpha diversity than CF, indicating enhanced selectivity in microbial enrichment. Overall, CF-HA achieved the highest methane yield (304.1 mL CH 4 /g COD), ~20% higher than the CF control. This study demonstrates that modification-specific microbial enrichment critically governs MEC performance, whereas the quantity of biomass adhesion to the electrode is not the determining factor.

World journal of microbiology & biotechnology • 2025

In this study, a two-chambered microbial electrolysis cell (MEC) was fabricated using a carbon cloth electrode, a Nafion-117 membrane, and Escherichia coli as the biocatalyst. The system treated sewage sludge with an initial chemical oxygen demand (COD) of 14,843 ± 350 mg/L, reducing it to approximately 170 ± 12 mg/L over the operational period. This configuration facilitated a significant increase in cell volume utilization of 89.5%, and achieved a maximum current production of 18.03 mA, along with hydrogen generation of 24.34 ± 1.3 mL at a biomass concentration of 13.5 ± 1.1 g/L. Cell growth contributed substantially to the degradation of organic matter, with sludge consumption reaching 12 g/L. The system produced an output voltage of 1.88934 ± 0.0091 V within 72 h and achieved the highest power density of 261,037 ± 1,245 mA/m 2 . The improved performance is attributed to the unique configuration of intermembranous live cells, highlighting the potential of biological electrocatalysts in advancing MEC technology. Further experiments demonstrated that when the initial COD load was 2,443 ± 175 mg/L, the MEC achieved a maximum COD removal efficiency of 98.5%. These findings emphasize the importance of optimizing sewage sludge substrate concentration to maximize power generation in MECs while simultaneously reducing the environmental burden of sludge biomass.

Bioresource technology • 2025

Volatile organic compounds (VOCs) severely impact human health and ecosystems, making effective removal technologies crucial. This study constructed a microbial electrolysis cell (MEC) using in situ synthesized nitrogen-doped carbon nanotube-modified carbon cloth (N-CNTs/CC) electrodes, which significantly enhanced VOCs degradation. N-CNTs reduced bioenergetic charge transfer impedance by 21.68 %, and their 3D conducting network, combined with electroactive microorganisms, promoted both direct and flavin-mediated indirect electron transfer. The system achieved 100 % removal of methyl methacrylate (MMA) within 6 h, 30.63 % higher than with carbon cloth electrodes. In mixed contaminants, MMA inhibited the degradation of other components. Meanwhile, selective enrichment of functional bacteria like Pseudomonas and Acinetobacter clarified microbial substrate response mechanisms. Cyclic voltammetry confirmed cytochrome-mediated direct electron transfer dominance. Under continuous flow with a 3-hour hydraulic retention time, complete MMA removal was achieved, showing strong engineering application potential. This study provides a theoretical basis for optimizing electrode strategies to advance energy-efficient waste treatment technologies.

Environmental research • 2025

Facing the issue of phosphorus (P) resource scarcity, microbial electrolysis cells (MECs) have garnered widespread attention from researchers due to their ability to recover P in the form of struvite without the need for external alkali addition. This study focuses on enhancing P recovery efficiency through modifying stainless steel mesh (SSM) cathodes with nickel-cobalt-tin (Ni-Co-Sn) composite materials and the circulation of catholyte in MECs. Results demonstrated that under optimal electrodeposition parameters (current density: 0.3 A/dm 2 , duration: 1800 s, temperature: 50 °C, and Ni:Co:Sn molar ion ratio of 2:1:0.5), the Ni-Co-Sn-SSM cathode system significantly reduced the charge transfer resistance of the original SSM, achieved a solution pH of 8.85 within 12 h, and exhibited exceptional P recovery efficiency of 87.5 %. In cyclic operation mode with controlled conditions, the P recovery efficiency reached 96.4 % within 8 h and attained near-complete recovery (99.9 %) after 12 h. The Ni-Co-Sn-SSM cathode enhances P recovery by maximizing hydrogen adsorption and desorption, increasing OH - generation, improving hydrogen evolution catalytic activity, and expanding the specific surface area to boost active sites and catholyte pH. Cathode modification and catholyte circulation significantly enhance P recovery, offering a sustainable solution for efficient P resource recycling.

Environmental monitoring and assessment • 2025

Anaerobic ammonium-oxidizing bacteria (AnAOB) present significant enrichment challenges due to slow growth kinetics and extended cultivation periods. This study employed single-chamber microbial electrolysis cells (MECs) to investigate substrate conversion rates, microbial community dynamics, and bacterial morphological adaptations in mixed consortia (incorporating nitrifiers, denitrifiers, and AnAOB) under varying applied voltages (0-1.2 V). Key results demonstrated that at 0.6 V, ammonium removal efficiency nearly doubled compared to the 0 V baseline, with AnAOB relative abundance increasing from 5.78 ± 0.38% to 12.34 ± 0.60% while denitrifiers decreased from 9.69 ± 1.33% to 2.84 ± 0.14%. Voltage escalation to 1.2 V induced proliferation of denitrifiers, which coexisted with AnAOB under carbon-limited conditions. During actual wastewater operation, ammonium degradation rates declined to 73% (Cycle 1) and 68% (Cycle 2), yet AnAOB maintained functional dominance at 12.56 ± 0.40% relative abundance. These findings provide valuable insights for AnAOB enrichment from complex communities, demonstrating voltage optimization's potential to enhance syntrophic interactions and intensify nitrogen removal pathways.

Bioresources and bioprocessing • 2025

Volatile fatty acid (VFA) accumulation is a common issue that compromises the performance of biological hydrogen methanation systems (BHMs). This accumulation is often triggered by fluctuations in hydrogen supply, which can disrupt microbial activity and lead to system instability. To address this challenge, this study investigated the impact of employing a microbial electrolysis cell (MEC) in BHMs to mitigate system instability and acid build-up. As such, a conventional anaerobic digester (AD) and a microbial electrolysis cell, both supplemented with exogenous hydrogen, were evaluated for their performance in hydrogen methanation. The effect of exogenous hydrogen at high addition rates (> 4:1 CO 2 :H 2 molar ratio) under instantaneous and gradual injection modes was investigated. The results showed that the instantaneous addition of hydrogen resulted in the total failure of the anaerobic digestion system. Propionate accumulated in the system (> 2 g/L) and resulted in low pH (pH = 5.3). Methane production stopped, and the reactor never recovered from hydrogen shock. However, the microbial electrolysis system was able to withstand the instantaneous hydrogen addition and maintain normal operation under toxic hydrogen addition levels (> 4:1 CO 2 :H 2 molar ratio). Under the gradual injection mode, both MEC and AD reactors remained reasonably unaffected; even though the hydrogen injection exceeded the stoichiometric molar ratio. This study provides a new perspective on the application of MECs for reliable operation and storage of surplus renewable energy via biological hydrogen methanation.

Bioresource technology • 2025

Microbial electrolysis cells (MECs) offer a sustainable approach to producing hydrogen (H 2 ) from organic waste. However, industrialization of MECs is hindered by a low H 2 production rate (HPR), mainly due to inefficient mass transfer between microbes and electrodes. To overcome this limitation, efficient dual-chamber MEC reactor design and evaluation of various substrate applicability are essential. In this study, we investigated the potential of mixed fruit waste (MFW) as an MEC substrate and its impact on microbial community dynamics in a multi-stack MEC reactor designed for H 2 production. To enhance microbial utilization, MFW was pretreated with eco-friendly organic acids (i.e., citric acid), achieving a high carbohydrate extraction efficiency of 82 %. Physicochemical analyses of MFW before and after pretreatment confirmed effective hydrolysis. MEC operation with 25 % MFW extract achieved a high chemical oxygen demand (COD) removal efficiency of 77 % and a maximum current density of 0.71 A/m 2 (16.36 A/m 3 ). Organic components in the MFW extract, including 5-hydroxymethylfurfural (HMF) and furan derivatives, were completely decomposed during MEC operation. Notably, microbial community analysis revealed distinct spatial distributions across the anode's vertical positions. Fermentative bacteria predominated in the bottom section, while electroactive genera such as Geobacter and Comamonas dominated the top section, likely due to the upward flow and recirculation of the organic substrate introduced at the bottom. The proposed multi-stack MEC process enhanced substrate utilization and microbial symbiosis interactions, highlighting its potential for industrial-scale applications.

Bioresource technology • 2026

Microbial electrolytic cell (MEC) can achieve autotrophic nitrogen removal, which is a promising means for treating low C/N ammonium-rich wastewater. However, it still suffers from insufficient ammonium oxidation rate and low total nitrogen (TN) removal. This study optimized the autotrophic nitrogen removal performance of a single-chamber MEC reactor using carbon granule (CG) materials as the bioanode, combined with supercapacitor (SC) as power supply. In the anoxic anodic system with only ammonium as the electron donor, the CG-anode/SC-powered system achieved removal loads of 35.31 ± 4.89 and 27.44 ± 4.80 mg L -1 d -1 for NH 4 + -N and TN, respectively. This TN removal load was 7 and 5.8 times higher than those using carbon felt and carbon brush anodes, respectively, and 2.7 times higher than those using a direct current power supply. Microbial community composition and functional gene abundance analyses showed that the CG anode was occupied by microbes associated with the anaerobic ammonia oxidation (Candidatus_Kuenenia, Ignavibacterium, norank__c__SJA-28, and norank__f__A4b), combined with the key gene abundance increase of hzs and hdh in the anammox pathway. With the lower current and coulomb efficiency of the CG-anode/SC-powered system, it demonstrated that the mechanism on the enhanced nitrogen removal was the electrically driven anodic partial nitrification with electrode as electron acceptor for nitrite production in situ, and then combined with anammox for nitrogen removal. This study constructs a novel autotrophic nitrogen removal system combined with SC-MEC and anammox, providing a viable strategy for the treatment of low C/N ammonium-rich wastewater.

World journal of microbiology & biotechnology • 2025

The growing demand for freshwater, coupled with the increasing volume of industrial and municipal wastewater, has intensified the need for sustainable and eco-friendly reclamation strategies. Recent advancements in microbial biotechnology have emerged as promising tools for developing cost-effective, efficient, and environmentally sustainable wastewater treatment (WWT) strategies for reuse and safe disposal. This mini-review explores current innovations, such as microbial consortia, bioaugmentation, and the microalgae-bacteria nexus, which have shown promising results in nutrient removal, enhanced degradation of complex pollutants (including emerging contaminants), and biomass valorization. Moreover, bioelectrochemical systems, such as microbial fuel cells (MFC) and microbial electrolysis cells (MEC), have revolutionized WWT by facilitating pollutant degradation while simultaneously generating bioelectricity or biohydrogen. This article also critically examines the role of CRISPR-based tools and 'omics' approaches, which have enabled the development of novel microbial strains and degradative pathways, enhancing wastewater reclamation in challenging environments. Furthermore, advancements through the integration of multi-omics and artificial intelligence, digital twins, and Internet of Things (IoT) for microbial optimization and real-time process control are discussed. The review highlights the role of microbial systems in resource recovery, supporting a circular economy by transforming wastewater into valuable bioresources. Additionally, this review addresses the major challenges and proposes future research directions for effective wastewater treatment. The novelty of this manuscript is that no single review explores the cutting-edge microbial biotechnologies for wastewater reclamation, uniquely integrating CRISPR-Cas genome editing, multi-omics analyses, and artificial intelligence-driven optimization to advance pollutant degradation and real-time process control in one place. This study concludes that by implementing multi-omics and artificial intelligence (AI)-driven optimization process for wastewater treatment can be effective towards wastewater treatment while simultaneously minimizing the environmental pollution.

Bioresource technology • 2026

Methane production in microbial electrolysis cell-anaerobic digestion (MEC-AD) systems is limited by inefficient interfacial electron transfer (IET), which depends on the slow formation of a conductive microbial biofilm. In this study, a novel material with Fe (III) ions as the metal centers and the redox-active 2,5-dihydroxy-1,4-benzoquinone (dhbq) as the organic ligand (Fe@dhbq) was designed to act as an efficient "electron pool" that mediated electron transfer between the cathode and electroactive methanogens, which helped establish an efficient IET network by accelerating the maturation of electroactive biofilms. The results demonstrated that the electron transfer capacity of the CC/Fe@dhbq biofilm increased by an order of magnitude, and the current was 4.2 times higher than the control. Correspondingly, direct electron transfer (DET) pathway was formed to increase methane production by 35 %. This work provides a new strategy for enhancing methanogenesis via DET in electro-enhanced anaerobic digestion.

Journal of environmental management • 2026

Addressing the challenges of unstable activity in microbial sulfate-reducing bacteria (SRB) and the need for further recovery of sulfate reduction products, this study employed iron-carbon (Fe-C) micro-electrolysis to enhance microbial sulfate reduction (MSR) and achieve sulfur resource recovery. By comparing the synergistic effects of iron (Fe), carbon (C), and Fe/C composite systems with a polyvinyl alcohol-sodium alginate (PVA-SA) immobilization system, the key mechanisms were elucidated. The optimized Fe-C system (Fe/C mass ratio 2:1, 6 g Fe/L: 3 g C/L) reduced sulfate from 1000 mg/L to 362.9 mg/L within 5 d, while minimizing sulfide accumulation to 48.6 mg/L via in-situ generation of iron sulfides (FeS). X-ray photoelectron spectroscopy (XPS) confirmed FeS formation, validating direct sulfur recovery. Micro-electrolysis selectively enriched completely oxidizing sulfate-reducing bacteria (SRB), particularly Desulfococcus (relative abundance: 10.5 % in Fe-C group vs. 3.7 % in control), enhancing metabolic efficiency. PVA-SA hydrogel immobilization significantly improved microbial stress resistance, manifested as increased cell viability (1.21 × control) and stabilized extracellular polymeric substances (EPS). This alleviated Fe 3+ cytotoxicity and maintained reducing conditions (ORP stabilized below -200 mV). Under the optimal Fe/C ratio, electron transfer system activity (ETSA) reached its maximum (1.4 × control). The research demonstrates that Fe-C micro-electrolysis combined with PVA-SA immobilization establishes an efficient platform for sulfate removal and sulfur resource recovery, providing a sustainable strategy for treating sulfate-laden wastewater.

Bioresource technology • 2026

Zero-gap microbial electrolysis cells (MECs) have demonstrated large current and hydrogen production rates from defined substrates in synthetic media, but operation with real waste streams has yet to be proved. This study evaluated the performance and 30-days stability of zero-gap MECs operated with effluent from a single-stage anaerobic digester. The system achieved a maximum current density of 8.8 ± 0.3 A/m 2 with a hydrogen production rate of 32 ± 6 L/L-d, and during 30 days of continuous operation, sustained an average current density of 7 ± 2 A/m 2 and a hydrogen production rate of 20.8 ± 0.2 L/L-d. Carbonate precipitation was identified as a major challenge to long-term stability, and mild acid washing effectively mitigated its adverse effects. The low buffer capacity of the effluent was primarily limiting performance. These findings underscore the significant impact of wastewater chemistry on MEC operation and validate the feasibility of utilizing real waste streams as viable feedstocks for biohydrogen production in zero-gap configurations.

Bioresource technology • 2026

This study presents an integrated approach combining microbial electrolysis cells (MECs) with three external ammonium extraction systems, Gas Diffuser (GD), Stripping Column (SP) and Thermally Assisted Stripping (SP + T), to enhance nitrogen removal and recovery from high-strength wastewater. Among the configurations, MECs operated at 0.75 V and integrated with SP + T achieved the highest performances, with current density of 8.8 A m -2 , ammonium removal and recovery rates reaching 69.8 and 37.9 gN m -2  d -1 , Faradaic efficiency of 74.9 % and specific energy consumption for ammonium recovery of 3.3 kWh kg -1 N. The SP + T system boosted ammonia volatilization at the catholyte and accelerated ammonium transport across the cation exchange membrane, leading to improved removal and recovery performance. Concurrently, H 2 generation reached 1557.6 L H 2  m -3  d -1 with specific energy consumption of 31.4 kWh kg -1 H 2 . These results highlight that coupling MEC and SP + T is a promising configuration for efficient nitrogen recovery and renewable hydrogen generation, while demonstrating MECs as a sustainable technology for wastewater treatment and resource recovery. STRUCTURED ABSTRACT: The study presents a novel approach for enhancing nitrogen recovery from wastewater using MECs integrated with different ammonium stripping systems. By evaluating three configurations, gas diffuser (GD), ambient-temperature stripping (SP) and thermally assisted MEC stripping system (SP + T), this work systematically investigates how operational strategies affect the performance of nitrogen removal and recovery. Key performance indicators such as current density, Faradaic efficiency, specific energy consumption and hydrogen production are quantified and compared as well. Furthermore, organic removal rates, percentage and energy efficiency of organic matter treatment in the anode are also assessed. The findings described highlight the importance of optimizing operational parameters such as temperature and contact surface to maximize ammonium recovery in MECs. The SP + T approach is relevant for bioelectrochemical and wastewater treatment systems aiming to optimize nitrogen management and valorization. Furthermore, the study includes a comprehensive energy analysis and contextualizes the results with respect to conventional ammonia production technologies such as the Haber-Bosch process, among others. Thus, these results are of relevance to the fields of wastewater treatment, nutrient recovery, energy/hydrogen production, environmental biotechnology and circular economy, offering a scalable and energy-efficient solution for nutrient valorization.

Scientific reports • 2026

The transition to a hydrogen-based energy system requires safe and efficient large-scale storage solutions. Underground hydrogen storage (UHS) has emerged as a promising candidate, but the effectiveness and long-term security of UHS depend on a fundamental understanding of hydrogen transport mechanisms in subsurface environments. Among these, hydrogen diffusion into formation water plays a key role in governing mass loss, geochemical reactions, and microbial activity. This study presents a combined experimental and mathematical approach to quantify hydrogen diffusion in distilled water and brine at temperatures of 30 °C, 45 °C, and 60 °C, and pressures near 6.5 MPa. A new formulation is proposed that accounts for non-constant gas compressibility and enables systematic identification of the initial non-diffusive regime in pressure-time data. The methodology was validated by measuring the diffusion coefficient of CO₂ in distilled water, giving values consistent with those reported in the literature. Experimental results showed that hydrogen diffusion coefficients increase with temperature, from 3.6[Formula: see text]0.4 × 10⁻⁹ m²/s at 30 °C to 6.1[Formula: see text]0.3 × 10⁻⁹ m²/s at 60 °C. An additional series of tests on brine demonstrated that increasing salinity from 0 to 10,000 ppm of NaCl salt reduces the hydrogen diffusion coefficient to 5.[Formula: see text]0.2 × 10⁻⁹ m²/s at 60 °C. This methodology offers a robust framework for characterizing gas diffusion in aqueous systems, providing critical input not only for modeling hydrogen behavior in geological storage formations but also for optimizing hydrogen production via water electrolysis and its utilization in fuel cells.

ACS sustainable chemistry & engineering • 2026

Microbial electromethanogenesis (EM) presents a promising pathway for sustainable biogas upgrading, but accurately predicting its performance is challenging due to complex, nonlinear process dynamics. Here, we systematically compared seven supervised machine learning (ML) algorithms, including one-dimensional convolutional neural network (1D-CNN), multilayer perceptron (MLP), gradient boosting regressor (GBR), adaptive boosting regressor (AdaBoost), stacking regressors, and K-nearest neighbors (kNN), for their predictive biomethane production capabilities using experimental data from EM bioelectrochemical systems (EM-BESs). The data set encompassed operational parameters such as optical density (OD 600 ), pH, electrical conductivity (EC, mS/cm), average applied current (A m -2 ), and CO 2 availability (mol). After hyperparameter optimization, the 1D-CNN model exhibited superior predictive performance ( R 2 = 0.934), significantly outperforming traditional ML methods. To move beyond prediction and uncover mechanistic insights, a feature importance analysis was conducted on the CNN model using SHapley Additive exPlanations (SHAP). The analysis revealed that average current, OD 600 , and pH were the most influential features in biomethane production, confirming that the model learned relationships grounded in fundamental bioelectrochemical principles. The SHAP analysis also identified complex, nonmonotonic effects of other variables, providing deeper process understanding. This study not only demonstrates the promising ability of ML, especially deep learning architectures, to advance EM optimization but also provides mechanistic insights into the factors governing bioelectrochemical methanogenesis. These findings are broadly applicable to analogous BESs, particularly microbial electrosynthesis (i.e., commodity chemical) and microbial electrolysis cells (i.e., biohydrogen), offering potential for enhancing system performance through data-driven operational control across sustainable biotechnology applications.

Bioresource technology • 2026

Microbial upcycling of heavy metal wastewater into catalysts offers a sustainable solution for antibiotic degradation, yet traditional synthesis is hindered by severe metal toxicity and complexation issues. To address these challenges, a stepwise biosynthesis strategy was developed using Shewanella oneidensis MR-1 to fabricate copper-manganese bimetallic sulfide (bio-CuMnSs) from heavy metal-laden wastewater. Unlike the one-step method, this strategy maintained high cell viability, enabling the efficient production of uniform nanoparticles. The bio-CuMnS (Cu/Mn molar ratio of 2:1) significantly outperformed its chemically synthesized counterpart, achieving 96 % degradation of tetracycline hydrochloride (TCH, 20 mg/L) within 60 min via peroxymonosulfate (PMS) activation. Mechanistic investigations revealed that the degradation was predominantly driven by surface-bound radicals (HO • /SO 4 •- ) confined by biogenic functional groups, with supplementary contributions from singlet oxygen ( 1 O 2 ). A distinct bimetal-sulfur synergy was confirmed, where intermediate-valence sulfur species (S 2 2- , S n 2- , and S 0 ) served as internal electron donors to drive the cooperative regeneration of Cu(I) and Mn(II) active sites, thereby sustaining PMS activation. The bio-CuMnS/PMS system maintained high efficiency over a broad pH range (5-11), in complex water matrices, and for different tetracycline antibiotics. Furthermore, the catalyst derived from real electroplating wastewater exhibited performance comparable to that from laboratory precursors, confirming the feasibility of simultaneous heavy metal valorization and antibiotic wastewater treatment.

Bioresource technology • 2026

Stable NO 2 - supply and low-temperature inhibition represent two major bottlenecks for anaerobic ammonia oxidation (anammox) in mainstream wastewater treatment. It was reported that applied electric fields enable anaerobic ammonium-oxidizing bacteria (AnAOB) to directly oxidize NH 4 + to N 2 . Whereas, low-temperature inhibition primarily stems from impaired electron transfer and subsequent metabolic suppression. Enhancing these processes through applied potential and conductive materials offers a strategy for this study. A single-chamber microbial electrolysis cell with 0.6 V anodic potential and 10 mg L -1 reduced graphene oxide (RGO) was developed. Results showed that the system achieved nitrogen removal rates of 7.4 ± 0.5g N m -3 d -1 and 74.2 ± 4.8 mg N m -2 d -1 at 10°C. NH 4 + -N and total nitrogen removal efficiencies increased by 116.6% and 147.0%, respectively. The applied potential upregulated pili/cytochrome expression, while the RGO network provided a low-resistance conductive matrix. As a result, synergic system effectively intensified electron transfer, which increased anode current density by 1.9-fold and reduced resistance by 67.7%, supporting both conventional anammox via enhanced nitritation and the electric-anammox pathway. Furthermore, it also promoted the enrichment functional genes, such as hzs and hdh (107.6%-238.1%), and enhanced cold adaptation marked by increased in extracellular proteins and polysaccharides (44.8%-18.9%). This work demonstrates coordinated electron transfer enhancement and metabolic activation through potential-RGO integration provides an innovative solution for energy-efficient nitrogen removal in cold-region wastewater treatment.

Bioprocess and biosystems engineering • 2026

Microbial bioelectrochemical technologies rely on the development of biofilms on electrode surfaces; therefore, a high surface area in packed anodes is advantageous for their performance. In addition, bioelectrochemical reactors (BERs) for hydrogen production require low-cost installation materials to enable large-scale implementation. In this study, a one-liter BER was constructed using 0.38 L of carbon felt as a packed bioanode, 0.65 L of compost leachate as the electrolyte, and a stainless-steel mesh cathode. The reactor was operated under an anode potential of 0.05 V vs. Ag/AgCl (KCl, 3.5 M) in batch cycles of 24 h each. After medium replacement, the maximum accumulated gas volume reached 2.37 L, corresponding to a production rate of 7.38 m⁻ 3 gas m⁻ 3 packed reactor d⁻ 1 . The cathode potential varied over time, leading to fluctuations in energy efficiency, which exceeded 100%. Average energy, cathode and coulombic efficiencies over eight operational cycles were 124 ± 64%, 118 ± 56%, and 120 ± 61%, respectively. The gas yield obtained from compost leachate in the BER was within the upper range of productivity reported for microbial electrolysis cells. This work demonstrates a sustainable alternative for BER installation and operation and proposes a monitoring strategy to track energy efficiency during hydrogen production.

Bioresource technology • 2026

Anaerobic ammonium oxidation (Anammox) provides a low-carbon pathway for nitrogen removal, yet its reliance on nitrite (NO 2 - -N) constrains large-scale application. Emerging evidence indicates that Anammox bacteria (AnAOB) can oxidize ammonium (NH 4 + -N) through extracellular electron transfer (EET) without NO 2 - -N. However, the long-term stability of this process in mixed communities remains unresolved. Here, microbial electrolysis cells were operated for 260 days to investigate how anodic potential and conductive carrier regulate EET-dependent Anammox. A threshold potential of 0.4-0.6 V (vs. SHE) enabled NO 2 - -N-free NH 4 + -N removal of 103.61 ± 9.22 mg N·L -1 ·d -1 (approximately 2.5-fold higher than highly enriched communities) via a hydroxylamine oxidoreductase-mediated pathway. The conductive carrier increased electron flux 4.9-fold, enhanced protein secretion, and stabilized biofilms. High potential combined with conductive carrier enriched electroactive AnAOB (Candidatus Kuenenia, Candidatus Brocadia) and induced a shift from NO 2 - -N-dependent to EET-dependent metabolism. These findings demonstrate sustained long-term EET-dependent Anammox and inform scalable, carbon-free nitrogen removal.

Water research • 2026

Microbial electrolysis cells (MECs) can produce green hydrogen while removing organic contaminants from liquid waste streams by leveraging the metabolic activity of electroactive microorganisms. Despite their potential in a sustainable, circular economy, large-scale MECs that can treat relevant volumes of wastewater have failed to deliver performance proportional to their lab-scale counterparts. The reason behind this lower performance at scale remains unclear. In this study, we developed a combined electrochemistry-, hydrodynamic-, and microbial ecology-informed framework to analyze and optimize MEC performance during scale-up, enabling accurate quantification of major limitations and the identification of strategies to overcome them, ultimately facilitating equivalent performance at scale. Applying this framework to the scale-up of a zero-gap MEC from 9 cm 2 electrode area to 100 cm 2 electrode area, resulted in similar maximum current densities in a 100 cm 2 MEC (21.7 ± 1.1 A/m 2 ) compared to a 9 cm 2 system (25.1 ± 2.7 A/m 2 ), as well as equivalent hydrogen production rates of 69.3 L/L-d (100 cm 2 ) and 67.7 ± 2.4 L/L-d (9 cm 2 ). COMSOL flow dynamics simulations were used to scale up the reactor configuration without negatively affecting electrolyte velocity and distribution in the cell, minimizing the increase in internal resistances during scale-up (11.7 ± 0.5 mΩm 2 at 9 cm 2 ; 19.7 ± 1.3 mΩm 2 at 100 cm 2 ). Microbial community structures were assessed at both scales using high-throughput sequencing, highlighting the differences of populations across electrode dimensions and operational parameters. The framework presented here accelerates the development of effective strategies toward the scale-up of MECs by furthering the understanding of how electrochemical, hydrodynamic, and microbial ecology parameters change as the reactor dimension is increased. Ultimately, this approach contributes to advancing electrochemical biotechnology toward practical deployment in energy-efficient wastewater treatment systems.

Veterinary and animal science • 2026

Livestock production significantly contributes to global greenhouse gas (GHG) emissions, particularly methane (CH₄), nitrous oxide (N₂O), and carbon dioxide (CO₂), posing challenges to climate change mitigation and environmental sustainability. This review explores advanced, system-wide approaches to reduce emissions from livestock systems while enhancing productivity, resilience, and resource efficiency. It covers short-term mitigation strategies such as dietary interventions-including methane inhibitors, microbial modulators, and natural compounds-that target enteric fermentation. Long-term solutions involve genetic and breeding innovations, such as microbiome-genome interaction analyses, CRISPR-based editing, and low-methane phenotyping, supported by genomic selection and precision phenotyping tools. The review also assesses advanced manure management technologies like anaerobic digesters and nutrient recovery systems, and examines precision livestock farming tools, including real-time sensors, machine learning models, UAVs, and IoT-based monitoring systems. Emerging digital tools, blockchain, augmented reality, and AI-assisted diagnostics are highlighted for enhancing traceability and decision-making. The potential of integrated energy systems, such as microbial fuel cells, hydrogen electrolysis, algae-based bioenergy, and thermal gasification, is discussed alongside traditional renewables, enabling livestock farms to become clean energy hubs. Circularity is emphasized through silvopasture, algal bioremediation, insect bioconversion, and integrated crop-livestock systems. Environmental assessment tools and the socio-political dimensions of technology adoption, including policy, education, and farmer behavior, are also considered. Future research directions, such as atmospheric methane oxidation, 4D-printed feed additives, and quantum modeling, are proposed. Overall, the review calls for a transdisciplinary, integrated approach to transform livestock systems into climate-smart, low-emission food production networks.

iScience • 2026

Specialized cytochrome protein complexes conduct electrons across cell membranes in electrogenic bacteria, which enables these microbes to be harnessed for applications in electrical generation, biosensing, and microbial electrosynthesis. Here, we engineer the surface-exposed MtrC subunit from the MtrCAB complex of Shewanella oneidensis to enable selective cell attachment to functional materials, including electrodes for improved bioelectricity production. Incorporating a SpyTag bioconjugation domain on MtrC enables specific covalent attachment of SpyCatcher-fused proteins to MtrCAB on S. oneidensis and Escherichia coli . Importantly, the MtrC modification does not disrupt electron export, offering opportunities to interface cells with electronic materials. In the second approach, incorporating a graphite binding sequence on MtrC improves S. oneidensis attachment to graphite electrodes, yielding 30% greater current production in a microbial electrolysis cell compared to a variant expressing unmodified MtrC. An engineerable platform on the surface of electrogenic cells creates numerous opportunities for biotic-abiotic interface manipulation.

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Research Square • 2026

Abstract Rapid expansion of battery-power-driven technologies in electric vehicles, autonomous systems, aerospace platforms, defensesystems and medical devices has strengthened the demand of energy storage systems for safe, reliable and fault-tolerantoperations. Lithium-ion batteries with high energy density and competent electrochemical properties, are characteristicallyvulnerable which leads to cell level degradation, imbalance, and even failure. Battery Packs of large scale are composed ofseveral cells (hundreds or thousands). The failure of a single cell in the large scale battery packs can significantly result indegradation of performance of the system. And in extreme cases it could also lead to the shutdown of complete system. Thepaper presents a comprehensive modeling of the battery module by incorporating redundant cell switching mechanism thatwhich provides a simulation framework which can act as an intelligent fault tolerant battery module using MATLAB and Simulink.The battery module of the battery pack is designed in such a way that it consists two primary cells and a redundant cell whichare interconnected and are controlled by switching network comprising nine electronically controlled switches. The controlalgorithm implemented to monitor the cell voltage, current, State of Charge (SOC) continuously and dynamically reconfiguresthe topology if the module considering the primary cell voltage reaching the predefined threshold value. The modular designenables scalable integration of such multiple modules to form a complete battery pack, offering enhanced reliability, faultisolation, and maintainability. The system proposed is validated in the simulation environment in detail and is followed bythe incorporation of cell balancing hence by demonstrating the suitability for applying in the advanced battery managementsystems where safety-critical applications play a vital role.

[object Object], [object Object], [object Object] et al.

Applied and Environmental Microbiology • 2026

ABSTRACT Antibiotic contamination from biogenic waste in agricultural soils poses a significant threat to soil health and crop productivity. We investigated the effect of antibiotics on the soil microbial community, antibiotic resistance genes, and mobile genetic elements (MGEs) and plant productivity in a 6-week greenhouse trial. Here, Spinacia oleracea (spinach) and Raphanus sativus (radish) were grown from seeds, and a mix of five antibiotics, namely sulfamethoxazole, trimethoprim, enrofloxacin, clarithromycin, and chlortetracycline, was added to the soil at concentrations of 0, 0.1, 1, and 10 mg kg −1 soil dry weight (c0, c0.1, c1, and c10, respectively). Overall, we found that the antibiotic treatments significantly impacted prokaryotic α-diversity and prokaryotic and fungal β-diversity. The relative abundance of human and plant pathogens did not increase under antibiotic exposure, but there was a significant reduction in plant growth-promoting bacteria. Moreover, the c10 treatment significantly increased the abundance of MGE intI1, indicative of horizontal gene transfer and sulfonamide resistance gene sul1, and significantly lowered radish biomass and nitrogen uptake, while spinach biomass and nitrogen uptake were unaffected. In summary, our study showed that antibiotic exposure significantly changed prokaryotic community diversity, while fungi remained largely unaffected. The reduction of plant growth-promoting bacteria may have a significant impact on soil nutrient cycling and crop productivity, but more research is needed to understand the long-term impact of these co-applied antibiotics on food production. Additionally, more studies are needed to understand the effect of antibiotics on realistic, field-scale conditions to fully understand the impact on environmental and human health. IMPORTANCE Agricultural soils are frequently contaminated with complex mixtures of antibiotics from various biogenic sources, yet we lack a clear understanding of their specific ecological impact. While many studies investigate antibiotics, they are often studied in pollution sources like manure, which contain confounding factors like heavy metals. To provide a mechanistic understanding of antibiotic-specific responses, we investigated the effects of a complex, five-antibiotic mixture on the soil-plant system, independent of other contaminants. As expected, antibiotics reduced prokaryotic diversity and increased the abundance of some genes related to antibiotic resistance. Additionally, antibiotic exposure reduced plant growth-promoting bacteria, which may have subsequent detrimental effects on plant and soil health. Moreover, we found that antibiotic exposure can reduce plant biomass and nitrogen uptake, but this is highly plant dependent. This research highlights the critical need to monitor antibiotic pollution due to its potential detrimental effect on plant health and alterations to the soil microbiome.

[object Object], [object Object], [object Object] et al.

Frontiers in Cell and Developmental Biology • 2026

Introduction Persistent endoplasmic reticulum (ER) stress impairs early embryonic development by inducing apoptosis through C/EBP homologous protein (CHOP). Toll-like receptor 4 (TLR4), traditionally recognized for its role in innate immunity, has recently emerged as a modulator of intracellular stress responses. Lipopolysaccharide (LPS), a natural TLR4 agonist derived from Gram-negative bacteria, elicits both pro-inflammatory and cytoprotective effects depending on the cellular context and dosage. This study aimed to elucidate the role of TLR4 signaling in the regulation of CHOP-mediated apoptosis during porcine preimplantation development under ER stress. Methods Porcine embryos were treated with tunicamycin (TM, 5 nM) to induce ER stress and co-treated with LPS (10 μM) to activate TLR4 signaling. Developmental competence was assessed by blastocyst formation rates, total cell number, and markers of apoptosis and autophagy. Results LPS treatment significantly improved blastocyst formation rates compared to TM groups (TM: 37.50 ± 4.77% vs. TM+LPS: 52.89 ± 4.86%). Consistent with this improvement, the total cell number per blastocyst was significantly restored by LPS co-treatment (Control: 55.63 ± 2.15 vs. TM: 38.61 ± 2.57; TM+LPS: 48.84 ± 0.83), confirming enhanced cell proliferation under ER stress conditions. LPS co-treatment markedly reduced CHOP protein expression and suppressed ATF4 expression, indicating alleviation of PERK-ATF4-CHOP signaling. Additionally, autophagy and apoptosis were attenuated, as evidenced by a significantly decreased LC3-II/LC3-I ratio and a reduced number of TUNEL-positive cells. Notably, TLR4 knockdown abolished these LPS-mediated protective effects, confirming the requirement of TLR4 in mitigating ER stress-induced damage. Conclusion These findings demonstrated that LPS-mediated TLR4 signaling suppressed CHOP-induced apoptosis and autophagy under persistent ER stress, thereby improving embryonic viability. This study provides novel mechanistic insights into the non-canonical role of TLR4 in early embryonic development and highlights its therapeutic potential for improving in vitro embryo culture systems.

[object Object], [object Object], [object Object] et al.

Frontiers in Science • 2026

Global wastewater production exceeds 359 billion m 3 annually, of which only 52% is treated, mostly in expensive and resource-consuming processes. Microbial electrochemical technologies (METs) offer a transformative approach to sustainable wastewater management by converting waste into valuable resources such as energy, clean water, and nutrients. They present a viable solution to the United Nations’ Sustainable Development Goal 6 (to ensure access to water and sanitation for all) by enhancing both sanitation and resource recovery. METs, including microbial fuel cells (MFCs) and microbial electrolysis cells (MECs), harness electrogenic microorganisms to oxidize organic matter, generating electric energy or producing energy carriers like hydrogen and methane. METs also enable recovery of nutrients, such as ammonium and phosphates, which are essential for agriculture, thereby closing resource loops in a circular economy. Despite their potential, challenges remain in scaling up METs for widespread application. Pilot-scale MFCs and MECs have demonstrated feasibility, achieving up to 90% chemical oxygen demand removal and producing electric power, methane, or hydrogen from wastewater. However, high capital costs, material limitations, and energy efficiency barriers hinder commercialization. Innovations in electrode design, modular configurations, and integration with existing wastewater treatment processes (e.g., anaerobic digestion, membrane bioreactors, or constructed wetlands) are advancing METs toward higher technology readiness levels (TRLs 4–8). Field applications, like a system for urine-based electricity generation in underserved regions, highlight METs adaptability and societal impact. The transition from laboratory to real-world implementation requires scaling, process integration, and further optimization to reduce costs and improve performance. By aligning with circular economy principles, METs can transform wastewater into a resource, contributing to energy security, environmental sustainability, and global sanitation goals. Future research should focus on scalable designs, economic viability, and interdisciplinary collaboration alongside understanding and optimizing the microbial “black box” to enable METs to transform previously unused wastewater streams into valuable resources with targeted applications.

[object Object], [object Object], [object Object] et al.

Cell Discovery • 2026

Abstract S-palmitoylation, a reversible post-translational modification regulates protein stability and cellular functions, yet its role in glutamine metabolism remains unclear. Here, we show that ZDHHC14 as the key palmitoyltransferase catalyzing ASCT2 palmitoylation at conserved Cys39 and Cys48 residues, promoting lysosomal degradation of this glutamine transporter, whereas ABHD17B functions as a depalmitoylase to stabilize ASCT2. Mechanistically, glutamine deprivation activates JNK1, which directly phosphorylates ZDHHC14 at Thr440 residue, triggering its degradation and thereby enhancing ASCT2 stability. Importantly, combination of JNK and ASCT2 inhibitors synergistically inhibits glutamine metabolism and tumor growth in vivo. These findings reveal a phosphorylation-palmitoylation axis linking JNK-mediated ASCT2 palmitoylation and glutamine metabolism, offering a potential therapeutic strategy for non-small cell lung cancer.

[object Object], [object Object], [object Object] et al.

Journal of Composites Science • 2026

The commercialization of proton-exchange-membrane fuel cells is constrained by the limitations of perfluorosulfonic acid membranes like Nafion, which suffer from high methanol crossover, humidity-dependent conductivity, high cost, and poor environmental sustainability. This review presents a comprehensive analysis of aquaporin-inspired chitosan/cellulose (AQP-CS) composite membranes as a transformative, bio-inspired alternative. The central design paradigm integrates a sustainable chitosan/cellulose matrix—which offers inherent mechanical stability, tunable proton conduction, and excellent fuel barrier properties—with biomimetic water channels engineered for selective hydration transport. This synergistic architecture aims to fundamentally decouple water management from proton conduction, directly addressing the core performance flaw of conventional membranes. The review is structured to explicitly trace the logical pathway from the foundational material properties of chitosan and cellulose to the functional requirements for integrating synthetic aquaporin-mimetic components. Experimental evidence from advanced chitosan composites, demonstrating proton conductivities up to 0.131 S cm−1 alongside drastically reduced methanol permeability, validates the potential of this approach. Consequently, AQP-CS composites establish a novel framework for developing next-generation fuel cell membranes that combine high performance with ecological design. However, key challenges in the stable integration of biomimetic channels, long-term operational durability, and scalable manufacturing must be resolved to enable practical deployment and mark a significant leap toward sustainable energy conversion technologies.

[object Object], [object Object], [object Object] et al.

Research Square • 2026

Abstract As the main by-product of Rana chensinensis product processing, Rana chensinensis skin has important development and utilization value because it is rich in a variety of bioactive components. In this study, Bacillus subtilis was used as the fermentation strain to explore the efficient preparation process and antioxidant activity of peptides from Rana chensinensis skin. The fermentation conditions were optimized by single factor experiment and response surface analysis, and the optimal parameters were determined as follows: inoculum volume was 3%, fermentation time was 16 h, and shaking speed was 190 rpm. Under these conditions, the peptide content of the fermentation product reached 71.46 ± 0.92 mg/mL, and the DPPH scavenging rate was 81.29 ± 1.03%, which were 1.77-fold and 4.71-fold higher than those before fermentation, respectively, and the antioxidant activity was significantly improved. The molecular weight distribution of peptides in fermentation liquor was further analyzed by high performance liquid chromatography. The results showed that the proportion of peptides with molecular weight less than 10,000 Da was up to 90%, and 50.54% of small peptides were concentrated in the range of less than 3000 Da. Subsequently, small molecular peptides were obtained by ultrafiltration centrifugation, which showed higher DPPH scavenging rate, and their key role in antioxidant activity was verified. This study realized the high-value utilization of Rana chensinensis skin, not only laid an experimental foundation for its application in various fields, but also provided a scientific basis for the sustainable utilization of Rana chensinensis skin.

[object Object], [object Object], [object Object] et al.

Frontiers in Marine Science • 2026

Introduction Picocyanobacteria from the genera Prochlorococcus and Synechococcus thrive across the globe in aquatic environments, have relatively small genomes, and have growth dynamics regulated by both viral interactions and abiotic conditions, making them excellent model organisms for exploring host-pathogencoevolution. Methods We developed and refined methods to sample and sequence cyanobacteria, cyanophages, and measured features of their abiotic environment. Results The protocol described herein can successfully discriminate large-cell eukaryotic organisms, but size fractionation of picocyanobacteria appears to be affected by the presence of free DNA, multicellular structures, and abundant tycheposons. Our preferred final protocol from this exploratory effort included a combination of in-line and single vacuum flask filtrations, which reduced filtration processing time by over threefold in some cases compared to other tested methods, such as a fully in-line sequence or in-site filtrations. We successfully extracted an average of approximately 400–1200 ng for all filter fractions, with some variations between kits. Discussion The protocol described herein can successfully discriminate large-cell eukaryotic organisms, but size fractionation of picocyanobacteria appears to be affected by the presence of free DNA, multicellular structures, and abundant tycheposons.

[object Object], [object Object], [object Object] et al.

Foods • 2026

High-temperature Daqu (HTD)’s quality determines the characteristics and yield of the Chinese sauce-aroma baijiu. However, winter production frequently encounters challenges such as fermentation instability and metabolic fluctuations, primarily stemming from complex, unmonitored microenvironmental changes within the HTD pile. This study established a closed-loop system linking the microenvironment, HTD quality, microbiome, and metabolome. Through continuous monitoring of the winter fermentation pile’s microenvironmental conditions and integrating multi-omics analyses, we revealed that CO2 concentration within fermentation piles is the core factor causing quality variations in HTD. By breaking the respiratory bottleneck formed by carbon dioxide (CO2) accumulation through the turning anaerobic stress can be alleviated, thereby driving metabolic succession. The study found that vertical CO2 concentration heterogeneity severely restricts the enrichment of aerobic core functional microbial communities such as the Bacillus species. This directly blocks key metabolic pathways including amino acid metabolism and energy supply via ABC transporters. Moreover, the specific accumulation of Amadori products further confirms that this low-temperature environment under CO2 stress causes the Maillard reaction to stall at intermediate stages. Consequently, this study proposes a steady-state control strategy centered on oxygen and CO2 gas characteristics. By actively regulating the gaseous microenvironment to eliminate metabolic heterogeneity, it provides theoretical support for standardizing traditional fermentation processes.

[object Object], [object Object], [object Object] et al.

Cell Death Discovery • 2026

Abstract The maintenance of immune homeostasis is critical for tissue health and longevity, yet the regulatory mechanisms linking immune modulation to aging remain poorly understood. Here we found that the transcription factor cAMP response element-binding protein (CREB), activated by JNK signaling in aging guts, transcriptionally suppresses peptidoglycan recognition protein SC2( PGRP-SC2) —a homolog of anti-inflammatory PGLYRP1–4 with amidase activity. 16S rRNA sequencing revealed that CREB modulates not only microbial load but also microbiota composition. Elevated CREB activity decreased the Firmicutes/Bacteroidetes (F/B) ratio—a hallmark of age-associated dysbiosis in animals. Genetic enhancement of PGRP-SC2 rescues age-related gut hyperplasia, microbiota imbalance, and lifespan shortening induced by overactivation of CREB or its coactivator CRTC. Notably, CREB’s regulation of PGRP-SC2 operates independently of canonical immune pathways such as Imd/Relish, revealing a previously unrecognized layer of immune modulation. Our findings establish CREB as a central player in age-associated immune dysregulation and propose targeting the CREB-PGRP-SC2 axis as a potential therapeutic strategy for mitigating gut aging and its systemic consequences.

[object Object], [object Object], [object Object] et al.

Biochemistry and Cell Biology • 2026

The latest research on epigenetics in health and disease was reported at the 10 th Canadian Epigenetics, Environment and Health Research Consortium (CEEHRC) annual meeting at Blue Mountain Resort in Ontario in October, 2024. Canadian and international researchers from a wide range of disciplines and career stages convened to engage in interdisciplinary discussions on the latest advances in epigenomics, promoting collaboration and the exchange of knowledge in this rapidly evolving field. The meeting emphasized a comprehensive understanding of epigenetic mechanisms-such as DNA methylation, histone modification, and chromatin accessibility-that regulate key biological processes, from embryonic development to disease progression and treatment resistance. The CEEHRC Annual Meeting offered valuable insights from fundamental biology and basic science, helping to elucidate the epigenetic foundations of development and laying the groundwork for translational research aimed at combating epigenetically driven diseases.

[object Object], [object Object], [object Object] et al.

Research Square • 2026

Abstract Metabolic reprogramming is a fundamental determinant of immune cell fate and effector function during inflammation, yet how distinct metabolic pathways regulate pathogenic lung macrophage activity and contribute to fibrotic remodeling remains incompletely understood. Here, we integrated single-cell RNA sequencing, lineage-specific genetic mouse models, and fate-mapping approaches to define the metabolic pathways that regulate the pro-fibrotic activity of lung macrophages. We found that such cells undergo pronounced aerobic glycolysis in the bleomycin-induced mouse model of pulmonary fibrosis. Myeloid cell-specific depletion of lactate dehydrogenase A (Ldha), but not of mitochondrial pyruvate carrier (Mpc), markedly attenuated lung fibrosis and improved survival. Mechanistically, enhanced glycolytic flux increased lactate production, which promoted histone lysine lactylation and facilitated Arginase 1 (Arg1) expression in lung macrophages, thereby driving fibrotic progression. Using monocyte fate-mapping, we further demonstrated that Arg1 expression is largely restricted to recruited monocyte-derived macrophages rather than lung-resident macrophages. Notably, selective deletion of Ldha in granulocyte-monocyte progenitors and their progeny was sufficient to suppress Arg1 expression and reduce fibrosis severity. In human lung samples, we observed significantly elevated expression of ARG1 and key glycolytic enzymes in patients with idiopathic pulmonary fibrosis, despite species-specific differences in the immune cell types expressing ARG1. Together, these findings identify an aerobic glycolysis–lactate–histone lactylation axis that regulates pro-fibrotic myeloid cell function and represents a potential therapeutic target in pulmonary fibrosis.

[object Object], [object Object], [object Object] et al.

Research Square • 2026

Abstract Deep-sea microorganisms comprise the Earth's largest and least explored microbiome, yet the vast majority remain uncultivated due to challenges of preserving in situ high hydrostatic pressure and preventing loss of viability and diversity during recovery, which limits our ability to explore their ecological functions and adaptive strategies. Here, we introduce DeepDrop, a microfluidics platform that enables high-throughput single-cell cultivation under pressures spanning the full ocean depth directly aboard research vessels, following direct colony formation via pipette-generated double emulsions. Applying to hadal samples, DeepDrop recovered 50% more microbial diversity than conventional high-pressure bulk cultivation, including rare taxa with streamlined genomes and distinctive genetic features associated with pressure adaptation. Combined metagenomic and transcriptomic analyses revealed that DeepDrop enriched pressure-adapted taxa carrying key stress-related genes and induced coordinated transcriptional reprogramming, characterized by upregulation of stress pathways and repression of motility. By integrating shipboard deployment, pressure-stable droplet cultivation, and efficient recovery, DeepDrop offers a powerful platform for accessing deep-sea microbial dark matter and illuminating microbial life strategies under extreme environmental constraints.