Treatment of domestic wastewater and extracellular polymeric substance accumulation in siphon-type composite vertical subsurface flow constructed wetland.

In this study, the siphon-type composite vertical flow constructed wetland (Sc-VSsFCW) was constructed with anthracite and shale ceramsite chosen as the substrate bed materials. During the 90-day experiment, typical pollutant removal effects of wastewater and extracellular polymeric substance (EPS) accumulation were investigated. Meanwhile, X-ray diffraction and scanning electron microscopy were used to examine the phase composition and surface morphology to analyze adsorptive property. Additionally, we evaluated the impact of siphon effluent on clogging and depolymerization by measuring the EPS components' evolution within the system. The findings reveal that both the anthracite and shale ceramsite systems exhibit impressive removal efficiencies for total phosphorus (TP), total dissolved phosphorus (TDP), soluble reactive phosphorus (SRP), chemical oxygen demand (COD), ammonium nitrogen (NH4 +-N), and nitrate nitrogen (NO3 --N). However, as the experiment progressed, TP removal rates in both systems gradually declined because of the saturation of adsorption sites on the substrate surfaces. Although the dissolved oxygen (DO) levels remained relatively stable throughout the experiment, pH exhibited distinct patterns, suggesting that the anthracite system relies primarily on chemical adsorption, whereas the shale ceramsite system predominantly utilizes physical adsorption. After an initial period of fluctuation, the permeability coefficient and porosity of the system gradually stabilized, and the protein and polysaccharide contents in both systems exhibited a downward trend. The study underscores that anthracite and shale ceramsite have good effectiveness in pollutant removal as substrate materials. Overall, the hydraulic conditions of the double repeated oxygen coupling siphon in the Sc-VSsFCW system contribute to enhanced re-oxygenation capacity and permeability coefficient during operation. The changes in EPS content indicate that the siphon effluent exerts a certain depolymerization effect on the EPS within the system, thereby mitigating the risk of biological clogging to a certain extent. PRACTITIONER POINTS: The system can still maintain good pollutant treatment effect in long-term operation. The re-oxygenation method of the system can achieve efficient and long-term re-oxygenation effect. The siphon effluent has a certain improvement effect on the permeability coefficient and porosity, but it cannot effectively inhibit the occurrence of clogging. The EPS content did not change significantly during the operation of the system, and there was a risk of biological clogging.

Understanding microbial biomineralization at the molecular level: recent advances.

Microbial biomineralization is a phenomenon involving deposition of inorganic minerals inside or around microbial cells as a direct consequence of biogeochemical cycling. The microbial metabolic processes often create environmental conditions conducive for the precipitation of silicate, carbonate or phosphate, ferrate forms of ubiquitous inorganic ions. Till date the fundamental mechanisms underpinning two of the major types of microbial biomineralization such as, microbially controlled and microbially induced remains poorly understood. While microbially-controlled mineralization (MCM) depends entirely on the genetic makeup of the cell, microbially-induced mineralization (MIM) is dependent on factors such as cell morphology, cell surface structures and extracellular polymeric substances (EPS). In recent years, the organic template-mediated nucleation of inorganic minerals has been considered as an underlying mechanism based on the principles of solid-state bioinorganic chemistry. The present review thus attempts to provide a comprehensive and critical overview on the recent progress in holistic understanding of both MCM and MIM, which involves, organic-inorganic biomolecular interactions that lead to template formation, biomineral nucleation and crystallization. Also, the operation of specific metabolic pathways and molecular operons in directing microbial biomineralization have been discussed. Unravelling these molecular mechanisms of biomineralization can help in the biomimetic synthesis of minerals for potential therapeutic applications, and facilitating the engineering of microorganisms for commercial production of biominerals.

Biological self-protection inspired engineering of nanomaterials to construct a robust bio-nano system for environmental applications.

Nanomaterials can empower microbial-based chemical production or pollutant removal, e.g., nano zero-valent iron (nZVI) as an electron source to enhance microbial reducing pollutants. Constructing bio-nano interfaces is critical for bio-nano system operation, but low interfacial compatibility due to nanotoxicity challenges the system performance. Inspired by microorganisms' resistance to nanotoxicity by secreting extracellular polymeric substances (EPS), which can act as electron shuttling media, we design a highly compatible bio-nano interface by modifying nZVI with EPS, markedly improving the performance of a bio-nano system consisting of nZVI and bacteria. EPS modification reduced membrane damage and oxidative stress induced by nZVI. Moreover, EPS alleviated nZVI agglomeration and probably reduced bacterial rejection of nZVI by wrapping camouflage, contributing to the bio-nano interface formation, thereby facilitating nZVI to provide electrons for bacterial reducing pollutant via membrane-anchoring cytochrome c. This work provides a strategy for designing a highly biocompatible interface to construct robust and efficient bio-nano systems for environmental implication.

Chromium (VI) removal by magnetite nanoparticles immobilized with extracellular polymeric substances extracted from Lysinibacillus sp. WH.

Magnetite nanoparticles (nano-Fe3O4) and nano-Fe3O4 immobilized with bacterial extracellular polymeric substances (EPSs) extracted from Lysinibacillus sp. WH (Fe3O4/bact) were comparatively studied for the removal of Cr (VI) ions from aqueous solution in batch study. The objectives were to explore the removal of Cr (VI) efficiency by nano-Fe3O4 and Fe3O4/bact under varying bacterial concentrations at a range of acidic pH. Results indicated that 150 ppm Cr (VI) could be effectively removed by 5 g/L of nano-Fe3O4 at pH 4, with the efficiency of 89.2 ± 12%. The equilibrium time, determined by a pseudo-second-order model (R2 = 0.9983), was after 5 h, indicating chemical adsorption. The Cr (VI) removal by the nano-Fe3O4 immobilized with bacterial EPS was effective and steady under a wide range of acidic conditions although bacterial EPS has an alkaline nature. Here, we are the first to demonstrate that Cr (VI) removal efficiency by different concentrations of EPS was not significantly different, suggesting EPS concentration is possibly not the most crucial factor to be optimized for Cr (VI) removal in the future. This study shows the potential application of nano-Fe3O4 immobilized with bacterial EPS for wastewater treatment. PRACTITIONER POINTS: The equilibrium time for magnetite nanoparticles to remove Cr (VI) is 5 h, suggesting chemical adsorption. The Cr (VI) removal efficiency of either magnetite nanoparticles or bacterial EPS is stable under a wide range of acidic conditions. Magnetite nanoparticles immobilized with bacterial EPS extracted from Lysinibacillus sp. WH has a potential application for Cr (VI) removal in wastewater.

Unveiling the common laws of extracellular polymeric substances (EPS) properties on short-chain fatty acids production from sludge by EPS disintegration pretreatment.

The production of short-chain fatty acids (SCFAs) from sludge is promising, but the efficiency and product quality often vary because of extracellular polymeric substances (EPS) characteristics and pretreatment principles. This study adopted specific EPS disintegration pretreatment to treat different types of sludge. By correlation coefficient matrix analysis and correlation dynamics change resolution, the intrinsic relationships between the nature of EPS and the production of SCFAs from sludge was unveiled. We demonstrate that tight-bound EPS (TB-EPS) is a principal carbon reservoir, positively impacting SCFAs yields, in the fermentation system with EPS as the main fermentation substrate, it can contribute about 29.2 % for SCFAs growth during fermentation. Conversely, TB-EPS exhibits a negative correlation during fermentation due to EPS-SCFAs interconversion, while loosely bound EPS (LB-EPS) correlates positively. Proteins and polysaccharides in TB-EPS, especially proteins, significantly enhance individual SCFAs yields, predominantly acetic, propionic, and isovaleric acids. The findings would provide a theoretical basis for developing pretreatments and process-control technologies aimed at improving SCFAs production efficiency and quality.

Sludge bound-EPS solubilization enhance CH4 bioconversion and membrane fouling mitigation in electrochemical anaerobic membrane bioreactor: Insights from continuous operation and interpretable machine learning algorithms.

Bound extracellular polymeric substances (EPS) are complex, high-molecular-weight polymer mixtures that play a critical role in pore clogging, foulants adhesion, and fouling layer formation during membrane filtration, owing to their adhesive properties and gelation tendencies. In this study, a novel electrochemical anaerobic membrane bioreactor (EC-AnMBR) was constructed to investigate the effect of sludge bound-EPS solubilization on methane bioconversion and membrane fouling mitigation. During the 150-days' operation, the EC-AnMBR demonstrated remarkable performance, characterized by an exceptionally low fouling rate (transmembrane pressure (TMP) < 4.0 kPa) and high-quality effluent (COD removal > 98.2 %, protein removal > 97.7 %, and polysaccharide removal > 98.5 %). The highest methane productivity was up to 38.0 ± 3.1 mL/Lreactor/d at the applied voltage of 0.8 V with bound-EPS solubilization, 107.6 % higher than that of the control stage (18.3 ± 2.4 mL/Lreactor/d). Morphological and multiplex fluorescence labeling analyses revealed higher fluorescence intensities of proteins, polysaccharides, total cells and lipids on the surface of the fouling layer. In contrast, the interior exhibited increased compression density and reduced activity, likely attributable to compression effect. Under the synergistic influence of the electric field and bound-EPS solubilization, biomass characteristics exhibited a reduced propensity for membrane fouling. Furthermore, the bio-electrochemical regulation enhanced the electroactivity of microbial aggregates and enriched functional microorganisms, thereby promoting biofilm growth and direct interspecies electron transfer. Additionally, the potential hydrogenotrophic and methylotrophic methanogenesis pathways were enhanced at the cathode and anode surfaces, thereby increasing CH4 productivity. The random forest-based machine learning model analyzed the nonlinear contributions of EPS characteristics on methane productivity and TMP values, achieving R² values of 0.879 and 0.848, respectively. Shapley additive explanations (SHAP) analysis indicated that S-EPSPS and S-EPSPN were the most critical factors affecting CH4 productivity and membrane fouling, respectively. Partial dependence plot analysis further verified the marginal and interaction effects of different EPS layers on these outcomes. By combining continuous operation with interpretable machine learning algorithms, this study unveils the intricate impacts of EPS characteristics on methane productivity and membrane fouling behaviors, and provides new insights into sludge bound-EPS solubilization in EC-AnMBR.

Characterization of emetic Bacillus cereus biofilm formation and cereulide production in biofilm.

Bacillus cereus is a well-known foodborne pathogen that can cause human diseases, including vomiting caused by emetic toxin, cereulide, requiring 105-108 cells per gram to cause the disease. The bacterial cells may be eliminated during processing, but cereulide can survive in most processing techniques due to its resistance to high temperatures, extreme pH and proteolytic enzymes. Herein, we reported dynamic processes of biofilm formation of four different types and cereulide production within the biofilm. Confocal laser scanning microscopy (CLSM) images revealed that biofilms of the four different types reach each stage at different time points. Among the extracellular polymeric substances (EPS) components of the four biofilms formed by the emetic B. cereus F4810/72 strain, proteins account for the majority. In addition, there are significant differences (p < 0.05) in the EPS components at the same stage among biofilms of different types. The time point at which cereulide was first detected in the four types of biofilms was 24 h. In the biofilm of B. cereus formed in ultra-high-temperature (UHT) milk, the first peak of cereulide appeared at 72 h. The cereulide content of the biofilms formed in BHI was mostly higher than that of the biofilms formed in UHT milk. This study contributes to a better understanding of food safety issues in the industry caused by biofilm and cereulide toxin produced by B. cereus.

Multiple drivers and mechanisms of solid-water interfacial interactions in sludge dewatering: Roles of polarity and molecular structure of extracellular polymeric substances.

Water occurrence states in sewage sludge, influenced by sludge physicochemical properties, are crucial for sludge dewaterability and have recently been regarded as a research hotspot. Here, the multifold characteristics of sludge flocs during hydrothermal treatment, including rheological properties, solid-water interfacial interactions, and the polarity distribution and molecular structure of extracellular polymeric substances (EPS), were systematically investigated, and the impact of these characteristics on sludge dewaterability was explored in depth. Hydrothermal treatment at 80 °C and 100 °C induced the conversion of free water into bound water, while an increase in temperature to 180 °C resulted in a significant decrease in bound water content, approximately 4-fold lower than at 100 °C. In addition to the conventional view of decreased sludge surface hydrophilicity at high temperatures, the decline in bound water was associated with the reduction in sludge apparent viscosity. XAD resin fractionation identified the hydrophobic/hydrophilic EPS (HPO-/HPI) ratio as an important factor determining water occurrence states. Especially, hydrolysis of HPI-related hydrophilic proteins and subsequent increase in HPO-related tryptophan-like substances played a dominant role in reducing sludge viscosity and facilitating the release of bound water. Protein conformational analysis revealed that the disruption of α-helix structures and disulfide bonds significantly reduced EPS water-holding capacity, providing strong evidence for the potential of targeting these dense structure units to enhance sludge dewaterability. These findings provide a holistic understanding of multidimensional drivers of water occurrence states in sludge, and guide directions for optimizing sludge treatment efficiency through EPS modification.

Little alginates synthesized in EPS: Evidences from high-throughput community and metagenes.

As a significant structure in activated sludge, extracellular polymeric substances (EPS) hold considerable value regarding resource recovery and applications. The present study aimed to elucidate the relationship between the microbial community and the composition and properties of EPS. A biological nutrient removal (BNR) reactor was set up in the laboratory and controlled under different solid retention times (SRT), altering microbial species within the system. Then EPS was extracted from activated and analyzed by chemical and spectroscopic methods. High-throughput sequencing and metagenomic approaches were employed to investigate bacterial community and metabolic pathways. The results showed that lower SRT with a higher abundance of the family-level Proteobacteria (27.7%-53.5%) favored EPS synthesis, while another dominant group Bacteroidetes (20.0%-32.6%) may not significantly affect EPS synthesis. Furthermore, the abundance of alginates-producing bacteria including Pseudomonas spp. and Azotobacter vinelandii was only 2.53%-6.76% and 1.98%-6.34%, respectively. The alginate synthesis pathway genes Alg8 and Alg44 were also present at very low levels (0.05‱-0.11‱, 0.01‱-0.02‱, respectively). Another important gene related to alginates operons, AlgK, was absent across all the SRT-operated reactors. These findings suggest an impossible and incomplete alginate synthesis pathway within sludge. In light of these results, it can be concluded that EPS does not necessarily contain alginate components.

Mechanisms underlying the detrimental impact of micro(nano)plastics on the stability of aerobic granular sludge: Interactions between micro(nano)plastics and extracellular polymeric substances.

Microplastics (MPs) and nanoplastics (NPs) present in wastewater can pose a negative impact to aerobic granular sludge (AGS). Herein, this study found that MPs and NPs (20 mg/L) deteriorated the sludge settleability and granule integrity, resulting in a 15.7 % and 21.9 % decrease in the total nitrogen removal efficiency of the AGS system, respectively. This was possibly due to the reduction of the extracellular polymeric substances (EPS) content. The subsequent analysis revealed that tyrosine, tryptophan, and humic acid-like substances in EPS exhibited a higher propensity for chemisorption and inhomogeneous multilayer adsorption onto NPs compared to MPs. The binding of EPS onto the surface of plastic particles increased the electronegativity of the MPs, but facilitated the aggregation of NPs through reducing the electrostatic repulsion, thereby mitigating the adverse effects of MPs/NPs on the AGS stability. Additionally, comprehensive analysis of the extended Derjaguin-Landau-Verwey-Overbeek theory indicated that the suppressed aggregation of microorganisms was the internal mechanisms contributing to the inadequate stability of AGS induced by MPs/NPs. This study provides novel insights into the detrimental mechanisms of MPs/NPs on the AGS stability, highlighting the key role of EPS in maintaining the structural stability of AGS when exposed to MPs/NPs.

Multi-dimensional perspectives into the pervasive role of microbial extracellular polymeric substances in electron transport processes.

During the process of biological treatment, most microorganisms are encapsulated in extracellular polymeric substances (EPS), which protect the cell from adverse environments and aid in microbial attachment. Microorganisms utilize extracellular electron transfer (EET) for energy and information interchange with other cells and the outside environment. Understanding the role of steric EPS in EET is critical for studying microbiology and utilizing microorganisms in biogeochemical processes, pollutant transformation, and bioenergy generation. However, the current study shows that understanding the roles of EPS in the EET processes still needs a great deal of research. In view of recent research, this work aims to systematically summarize the production and functional group composition of microbial EPS. Additionally, EET pathways and the role of EPS in EET processes are detailed. Then factors impacting EET processes in EPS are then discussed, with a focus on the spatial structure and composition of EPS, conductive materials and environmental pollution, including antibiotics, pH and minerals. Finally, strategies to enhance EET, as well as current challenges and future prospects are outlined in detail. This review offers novel insights into the roles of EPS in biological electron transport and the application of microorganisms in pollutant transformation.

Removal of dibutyl phthalate (DBP) by bacterial extracellular polymeric substances (EPS) via enzyme catalysis and electron transmission.

Phthalic acid esters (PAEs) showed high environmental risk due to the widely existence and toxicity. Microbial-excreted extracellular polymeric substances (EPS) showed potential of degrading organic compounds. In this study, the degradation ability and the mechanisms of EPS from two bacteria (PAEs degrader Gordonia sihwensis; electrochemically active strain Shewanella oneidensis MR-1) were investigated. Results showed that EPS of the two bacteria had different composition of C-type cytochromes, flavins, catalase, and α-glucosidase. The removal of dibutyl phthalate (DBP) by total EPS were 68% of G. sihwensis and 72% for S. oneidensis. For both bacteria, the degradation rates k of EPS were as TB-EPS > LB-EPS > S-EPS. The degradation mechanisms of EPS from the two bacteria showed difference with electrochemical active components mediated electron transmission for S. oneidensis MR-1 and enzymes catalysis for G. sihwensis. Results of this study illustrated the variation of the contribution of active components of EPS to degradation.

The effect of salinity on trimethoprim adsorption by activated sludge extracellular polymeric substances at trace concentration.

The saline wastewater produced in industrial activities and seawater use would flow into wastewater treatment plants and affect the characteristic of extracellular polymeric substance (EPS) of activated sludge, which could potentially impact the removal of antibiotics via adsorption. Nonetheless, the effect of salinity on trimethoprim adsorption by activated sludge extracellular polymeric substances at trace concentration and the underlying mechanism remain largely unknown. In this study, the effect of salinity on the adsorption removal of a typical antibiotic, i.e., trimethoprim (TMP) at trace concentration (25.0 µg/L) was evaluated. The results showed the content of EPS was decreased significantly from 56.36 to 21.70 mg/g VSS when the salinity was increased from 0 to 10 g/L. Protein fractions occupied the predominant component of EPS, whose concentration was decreased from 38.17 to 12.83 mg/g VSS. The equilibrium adsorption capacity of activated sludge for TMP was decreased by 49.70% (from 4.97 to 2.50 µg/g VSS). The fluorescence quenching results indicated the fluorescence intensity of tryptophan-like substances was decreased by 30% and the adsorption sites of EPS were decreased from 0.51 to 0.21 when the salinity was increased. The infrared spectrum and XPS results showed that the nitrogen-containing groups from protein were decreased significantly. The circular dichroic analysis showed α helix structure of protein in EPS was decreased with the increase of salinity, which was responsible for the decrease of adsorption capacity for TMP.

Adaptation of the anammox process for high ammonium photovoltaic wastewater treatment.

Tunnel Oxide Passivating Contacts (TOPcon) battery in the photovoltaic industry generates high ammonium wastewater during the production process, the adaptability of using the anaerobic ammonia oxidation (Anammox) process for photovoltaic wastewater (PVW) treatment is a research hotspot. Based on the analysis of photovoltaic wastewater quality, the effectiveness of nitrogen removal, sludge characteristics and microbial communities were examined. The results showed that when the influent NH4+-N concentration of PVW was lower than 150 mg·L-1, the nitrogen removal efficiency (NRE) was almost 100 %. In addition, the NRE decreased from 74 % sharply to 20 % when the NH4+-N concentration was increased from 175 mg·L-1 to 200 mg·L-1. The extracellular polymeric substances (EPS) content increased with elevated ammonium concentration in the influent, indicating that microorganisms secreted more EPS to resist elevated nitrogen loading. The main functional populations were Candidatus Kuenenia (0-24 %). The influent ammonium concentration is recommended to be < 200 mg·L-1.

Rethinking characterization, application, and importance of extracellular polymeric substances in water technologies.

Biofilms play important roles in water technologies such as membrane treatments and activated sludge. The extracellular polymeric substances (EPS) are key components of biofilms. However, the precise nature of these substances and how they influence biofilm formation and behavior remain critical knowledge gaps. EPS are produced by many different microorganisms and span multiple biopolymer classes, which each require distinct strategies for characterization. The biopolymers additionally associate with each other to form insoluble complexes. Here, we explore recent progress toward resolving the structures and functions of EPS, where a shift towards direct functional assessments and advanced characterization techniques is necessary. This will enable integration with better microbial community and omics analyses to understand EPS biosynthesis pathways and create further opportunities for EPS control and valorization.

Variation of viscoelastic properties of extracellular polymeric substances and their relation to anaerobic granule's mechanical strength in full-scale treatment plants.

Extracellular polymeric substances (EPS) are considered to play a pivotal role in shaping granules' physical properties. In this contribution, we characterized the viscoelastic properties of EPS from granules of 9 full-scale industrial anaerobic reactors; and quantitatively investigated whether these properties correlate with granules' resistance to compression (Egranule) and shear strength (Sgranule). Most granules with a higher shear strength, also exhibited a stronger resistance to compression (r = 0.96, p = 0.002), except those granules that contained relatively more proteins in their EPS. Interestingly, these granules were also the most resistant to shear stress (Sgranule ≥ 110 ± 40 h). Furthermore, the EPS hydrogels of these granules had slower softening rates (κ < 0.9) compared to the others (κ ranged between 0.95 and 1.20), indicating stronger gels were formed. These findings suggest that the EPS hydrogel softening rate could be a key parameter to explain granule's shear strength.

Mixed species biofilm: Structure, challenge and its intricate involvement in hospital associated infections.

Hospital associated infections or healthcare associated infections (HAIs) are a major threat to healthcare and medical management, mostly because of their recalcitrant nature. The primary cause of these HAIs is bacterial associations, especially the interspecies interactions. In interspecies interactions, more than one species co-exists in a common platform of extracellular polymeric substances (EPS), establishing a strong interspecies crosstalk and thereby lead to the formation of mixed species biofilms. In this process, the internal microenvironment and the surrounding EPS matrix of the biofilms ensure the protection of the microorganisms and allow them to survive under antagonistic conditions. The communications between the biofilm members as well as the interactions between the bacterial cells and the matrix polymers, also aid in the rigidity of the biofilm structure and allow the microorganisms to evade both the host immune response and a wide range of anti-microbials. Therefore, to design a treatment protocol for HAIs is difficult and it has become a growing point of concern. This review therefore first aims to discuss the role of microenvironment, molecular structure, cell-cell communication, and metabolism of mixed species biofilms in manifestation of HAIs. In addition, we discuss the electrochemical properties of mixed-species biofilms and their mechanism in developing drug resistance. Then we focus on the most dreaded bacterial HAI including oral and gut multi-species infections, catheter-associated urinary tract infections, surgical site infections, and ventilator-associated pneumonia. Further, we highlight the challenges to eradication of the mixed species biofilms and the current and prospective future strategies for the treatment of mixed species-associated HAI. Together, the review presents a comprehensive understanding of mixed species biofilm-mediated infections in clinical scenario, and summarizes the current challenge and prospect of therapeutic strategies against HAI.

Role of diffusible signal factor in regulating aerobic granular sludge formation under temperature shocks.

Signal-molecule-mediated strategies are proposed for aerobic granular sludge (AGS), but the regulatory mechanisms behind AGS formation are largely unexplored. In this study, two sequence batch reactors (SBRs) were operated to investigate the regulation of diffusible signal factor (DSF) in AGS formation. DSF secretion in Reactor 2 (R2: 10 °C→25 °C) decreased by 15 % compared to Reactor 1 (R1: 25 °C→10 °C), correlating with a 26 % increase in extracellular polymeric substance (EPS) concentration, resulting in a 63 % acceleration of the granulation process. After temperature shocks in R2, DSF concentration increased by 70 %, while EPS concentration decreased by 47 %. Batch tests confirmed that DSF inhibited EPS secretion. Combined 16S rRNA analysis and machine learning identified key bacteria responsible for secreting EPS and signal molecule. The decrease in the abundances of these bacteria reduced EPS production. These findings on DSF regulation of EPS secretion provide an in-depth understanding of enhanced AGS granulation.

Accelerated healing of intractable biofilm-infected diabetic wounds by trypsin-loaded quaternized chitosan hydrogels that disrupt extracellular polymeric substances and eradicate bacteria.

Complex and stubborn bacterial biofilm infections significantly hinder diabetic wound healing and threaten public health. Therefore, a dressing material that effectively clears biofilms and promotes wound healing is urgently required. Herein, we introduce a novel strategy for simultaneously dispersing extracellular polymeric substances and eradicating drug-resistant bacteria. We prepared an ultrabroad-spectrum and injectable quaternized chitosan (QCS) hydrogel loaded with trypsin, which degrades biofilm extracellular proteins. Increased temperature initiated QCS gelation to form the hydrogel, enabling the sustained release of trypsin and effective adherence of the hydrogel to irregularly shaped wounds. To reproduce clinical scenarios, biofilms formed by a mixture of Staphylococcus aureus (S. aureus), Methicillin-resistant S. aureus, and Pseudomonas aeruginosa were administered to the wounds of rats with streptozotocin-induced diabetes. Under these severe infection conditions, the hydrogel efficiently suppressed inflammation, promoted angiogenesis, and enhanced collagen deposition, resulting in accelerated healing of diabetic wounds. Notably, the hydrogel demonstrates excellent biocompatibility without cytotoxicity. In summary, we present a trypsin-loaded QCS hydrogel with tremendous clinical applications potential for the treatment of chronic infected wounds.

Operation parameters and temperature affected sludge microbial metabolisms: An integrated perspective considering extracellular polymeric substances, soluble microbial products, biomass quantities, and community shifts.

Effluent soluble microbial products (SMP) and extracellular polymeric substances (EPS) are significant organics that pose challenges to advanced treatment processes. However, their production, transformation, and decomposition remain unclear due to their heterogeneity and the combined effects of environmental and operational factors. In this work, we investigated the impact of solids retention time (SRT), hydraulic retention time (HRT), and temperature on the changes in effluent SMP, with the consideration of the co-variation of EPS, sludge biomass, and community structures. Results show that longer SRT increased the biomass and relative abundance of functional microorganisms such as Myxococcota, Actinobacteria, and Terrimonas, which hindered EPS-to-SMP turnover and/or facilitated SMP consumption. This resulted in the accumulation of EPS and lower SMP concentrations at the beginning of the SRT adjustment. Both longer and shorter HRT (12 h and 8 h) led to increased SMP concentration, with the shorter HRT nearly doubling it (from approximately 6 to 12 mg/L), especially in terms of its protein and polysaccharide contents. Lower temperatures increased the SMP concentration and the relative abundance of Proteobacteria (including Zoogloea, the most dominant phylum and genus, relative abundance from 15.7 % to 61.1 %) while decreasing fluorescent EPS components, indicating the key role of Proteobacteria in SMP production and fluorescent EPS-to-SMP transformation. The results provided key insights into how changes in operational/environmental parameters impact sludge-EPS-SMP interactions, which could benefit the model development and operational optimization of activated sludge systems. This study also highlighted the important role of the sludge community in the EPS/SMP dynamics.