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Microbial biofilms exhibit a self-produced matrix of extracellular polymeric substances (EPS), including polysaccharides, proteins, extracellular DNA and lipids. EPS promote interactions of the biofilm with other cells and sorption of organics, metals and chemical pollutants, and they facilitate cell adhesion at interfaces and ensure matrix cohesion. EPS have roles in various natural environments, such as soils, sediments and marine habitats. In addition, EPS are relevant in technical environments, such as wastewater and drinking water treatment facilities, and water distribution systems, and they contribute to biofouling and microbially influenced corrosion. In medicine, EPS protect pathogens within the biofilm against the host immune system and antimicrobials, and emerging evidence suggests that EPS can represent potential virulence factors. By contrast, EPS yield a wide range of valuable products that include their role in self-repairing concrete. In this Review, we aim to explore EPS as a functional unit of biofilms in the environment, in technology and in medicine., Competing Interests: Competing interests: The authors declare no competing interests., (© 2024. Springer Nature Limited.)

Staphylococcus species, traditionally associated with pathogenicity, are gaining attention for their role in environmental bioremediation, particularly nitrate reduction, which is crucial for mitigating eutrophication. In this study, denitrifying, biofilm-forming Staphylococcus strains were isolated from Dal Lake, India. Biofilm formation was quantified using a microtiter plate assay, and extracellular polymeric substances (EPS) were measured by dry weight. Statistical analysis revealed a strong positive correlation between EPS production and nitrate removal efficiency (r = 0.96, p < 0.001), with EPS accounting for 92% of the variance in nitrate reduction (R 2  = 0.92). Among the isolates, Staphylococcus epidermidis exhibited the highest nitrate reduction at 87% (SD = 2.3%), followed by S. succinus at 83% (SD = 2.1%), S. equorum at 77% (SD = 2.5%), and Staphylococcus sp. at 70% (SD = 2.8%). The consistency of these findings was confirmed by boxplot analysis, and the regression model's robustness was validated by residual plots showing minimal systematic error. This research work provides the first evidence of the nitrate-reducing capabilities of these Staphylococcus species, underscoring their potential in sustainable bioremediation strategies for aquatic environments. The significant correlation between EPS production and nitrate reduction highlights the critical role of biofilms in enhancing microbial remediation processes. The study not only advances the understanding of Staphylococcus in non-pathogenic roles but also suggests that these strains could be pivotal in bioremediation technologies, potentially influencing future environmental management practices., Competing Interests: Declarations. Conflict of interest: The authors declare no competing interests., (© 2024. The Author(s), under exclusive licence to Springer Nature B.V.)

The biofilm matrix can be considered to be a shared space for the encased microbial cells, comprising a wide variety of extracellular polymeric substances (EPS), such as polysaccharides, proteins, amyloids, lipids and extracellular DNA (eDNA), as well as membrane vesicles and humic-like microbially derived refractory substances. EPS are dynamic in space and time and their components interact in complex ways, fulfilling various functions: to stabilize the matrix, acquire nutrients, retain and protect eDNA or exoenzymes, or offer sorption sites for ions and hydrophobic substances. The retention of exoenzymes effectively renders the biofilm matrix an external digestion system influencing the global turnover of biopolymers, considering the ubiquitous relevance of biofilms. Physico-chemical and biological interactions and environmental conditions enable biofilm systems to morph into films, microcolonies and macrocolonies, films, ridges, ripples, columns, pellicles, bubbles, mushrooms and suspended aggregates - in response to the very diverse conditions confronting a particular biofilm community. Assembly and dynamics of the matrix are mostly coordinated by secondary messengers, signalling molecules or small RNAs, in both medically relevant and environmental biofilms. Fully deciphering how bacteria provide structure to the matrix, and thus facilitate and benefit from extracellular reactions, remains the challenge for future biofilm research., (© 2022. Springer Nature Limited.)

Plastic pollution in aquatic environments poses significant concerns due to its potential to serve as a refuge for aquatic pathogens. However, the role of plastic surfaces and microbial biofilm interfaces in facilitating pathogen development remains poorly understood. In this study, a microcosm setup was employed to investigate the interactions between plastics and the microbial community and examine the differences in bacterial community composition and potential pathogen occurrences between the plastisphere-biofilm and surrounding seawater. Community composition analysis combined with SEM observations over time indicated that biofilm extracellular polymeric substance formation over 14 days had a link with the relative abundance and succession patterns of pathogen taxa. Colony clusters were observed on biofilms from day 7 and coincided with higher bacterial pathogen dominance. On day 14, pathogen abundance overall decreased with a potentially degrading biofilm. Pseudomonas and Pseudoalteromonas were the dominant potential pathogen groups observed in the microcosm. When further subjected to chemical treatment as an imposed environmental stress over time, biofilm-associated Psuedoalteromonas sharply increased in abundance after three days of exposure, but quickly diminished by 14 days in favor of genera such as Acinetobacter, Pseudomonas, and Staphylococcus. These results suggest that environmental plastisphere-biofilms can promote the early selection, enrichment, and spread of pathogenic bacteria in the aquatic environment and could be later worsened under chemical and long-term pressure. This study provided new insights into the succession of pathogens in plastisphere biofilms, contributing to the understanding of pathogen risks involved in emerging plastisphere biofilms in light of global plastic pollution., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Biofilm secreted by microalgae are extracellular polymeric substances (EPSs) composed mainly of polysaccharides, proteins, nucleic acids and lipids. These EPSs immobilize the cells and stabilize biofilm, mediating adhesion towards solid surfaces. The EPSs valorization through industrial exploitations and scientific works is becoming more popular, but the bottleneck of such studies is the lack of consensus among researchers on the selection of detection techniques to be used, especially for novice researchers. It is a daunting task for any inexperienced researcher when they fail to identify the right tools needed for microalgal biofilm studies. In this review, a well-refined analysis protocol about microalgal biofilm and EPSs were prepared including its extraction and characterization. Pros and cons of various detection techniques were addressed and cutting-edge methods to study biofilm EPSs were highlighted. Future perspectives were also presented at the end of this review to bridge research gaps in studying biofilm adhesion via EPSs production. Ultimately, this review aims to assist novice researchers in making the right choices in their research studies on microalgal biofilms in accordance to the available technologies and needs., (© 2022 Society for Applied Microbiology.)

Objectives: To test the role of exopolysaccharide (EPS) polymers matrix to modulate the composition/virulence of biofilms growing on titanium (Ti) surfaces, the effect on antibiotic susceptibility, and whether a dual-targeting therapy approach for disrupted EPS matrix could improve the antimicrobial effect., Materials and Methods: A microcosm biofilm model using human saliva as inoculum was used, and the microbial composition was assessed by checkerboard DNA-DNA hybridization. EPS-enriched biofilms virulence was tested using fibroblast monolayer. Povidone-iodine (PI) was used as EPS-targeting agent followed by amoxicillin + metronidazole antibiotic to reduce bacterial biomass using an in situ model., Results: An EPS-enriched environment, obtained by sucrose exposure, promoted bacterial accumulation and led to a dysbiosis on biofilms, favoring the growth of Streptococcus, Fusobacterium, and Campylobacter species and even strict anaerobic species related to peri-implant infections, such as Porphyromonas gingivalis and Tannerella forsythia (~3-fold increase). EPS-enriched biofilm transitioned from a commensal aerobic to a pathogenic anaerobic profile. EPS increased biofilm virulence promoting higher host cell damage and reduced antimicrobial susceptibility, but the use of a dual-targeting approach with PI pre-treatment disrupted EPS matrix scaffold, increasing antibiotic effect on in situ biofilms., Conclusion: Altogether, our data provide new insights of how EPS matrix creates an environment that favors putative pathogens growth and shed light to a promising approach that uses matrix disruption as initial step to potentially improve implant-related infections treatment., (© 2020 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.)

Understanding the impacts and mechanisms of different antibiotics on organic phosphorus (OP) mineralization is crucial for promoting the rational utilization of resources and protecting the ecological environment. In this study, the effects of four commonly used antibiotics (sulfadiazine, ciprofloxacin, erythromycin, and ampicillin) on the mineralization of OP were explored using16S rRNA gene sequencing technology. The results showed that ciprofloxacin, erythromycin, and ampicillin negatively affected the mineralization capacity of OP, whereas sulfadiazine positively influenced OP mineralization. The content and composition of extracellular polymeric substances (EPS) and microbial phenotypes (biofilm formation and stress tolerance) were directly correlated with differences in OP mineralization capacity. Microbial diversity, network complexity and stability, and key microorganisms indirectly influenced OP mineralization by regulating EPS content and composition and microbial phenotypes. In summary, this study reveals specific impacts of different antibiotics on OP mineralization, offering valuable insights for addressing "phosphorus limitation" and "phosphorus load" amid various antibiotic stresses., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier Ltd.)

Anammox granules harbor a variety of bacteria, with each granule layer providing a niche for bacteria with specific metabolic functions. To investigate microbial distribution, nitrogen transformation rates, and extracellular polymeric substance (EPS) content across the depth layers of anammox granules, granules sized 0.9-2.0 mm were sheared into seven layers. The outer layers exhibited higher anammox bacterial abundance (9.9 %) than the inner layers (8.1 %). In contrast, the abundance of denitrifying bacteria increased from 25.0 % in outer layers to 34.9 % in inner layers. Nitrifying bacteria, along with heterotrophic nitrifying and aerobic denitrifying bacteria, were predominantly found in outer layers. Despite the higher EPS content in inner layers, denitrification rates remained low across all layers, likely due to limited carbon availability from microbial lysis or EPS. These findings offer valuable insights into the niche distribution of microbial communities within anammox granules., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Extracellular polymeric substances (EPS) significantly influence the properties and performance of waste activated sludge. Various pretreatment protocols with different extraction efficiency and characteristics of EPS have been reported, which markedly impact subsequent treatment and disposal of sewage sludge. This study systematically assesses the EPS properties from twelve extraction pretreatment methods. The organic and inorganic matters content, cell lysis, basic physicochemical property, molecular weight distribution, and fluorescence properties of extracted EPS were determined. Physical extraction methods (Centrifugation, Heat, and Ultrasound) were relatively mild, resulting in lower extraction of organic matter contents from EPS (< 6 mg total organic carbon /g volatile solids). Biological extraction methods (Enzyme and Enzyme-NaOH) exhibited high EPS extraction efficiency however led to significant cell lysis, thereby contaminating the extracted EPS with intracellular substances. In addition, heating and biological extraction methods excessively degraded EPS, resulting in a large amount of small molecular weight matters (≤ 10 3  Da) generation. Chemical extraction methods offered efficient EPS extraction with the different degrees of cell lysis, but it would introduce chemical reagent residues in EPS. Among those extraction methods, EPS extracted by cation exchange resin (CER) method had uniform and abundant molecular weight matters and fluorescence matters distribution. After dialysis, the residual Na and other metal elements could be significantly removed. Then, the fluorescence quenching effect of Cu(II) and Zn(II) at 400 μM on the Bound-EPS after dialysis increased up to the maximum value of 65 % and 30 %, respectively, compared to that without dialysis. It indicates that dialysis coupled with CER extracted EPS has a well extraction efficiency and can maintain the original state of EPS for the subsequent investigation., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Disinfection by-products (DBPs), formed from biofilm extracellular polymeric substances (EPS) and organic matter during regular disinfection practices in drinking water distribution systems, poses a potential threat to drinking water safety. However, the diverse DBP formations induced by the intertwined algal organic matter (AOM) and bacterial EPS remains elusive. In this study, we show substantial variations in EPS and DBP formation patterns driven by AOM biosorption with divalent ions (Ca 2+ and Mg 2+ ). Divalent ions in bulk water can significantly inhibit carbonaceous DBPs (C-DBPs) and nitrogenous DBPs (N-DBPs) formation. Mechanistically, divalent ions promote the complexation of negative charged groups and thus inhibit C-DBP formation, while the hindering chlorine substitution of hydrogen atoms on α‑carbon and amine groups reduces N-DBP formation. Conversely, Ca 2+ and Mg 2+ could facilitate biosorption processes that increased the yields of C-DBPs and N-DBPs. Both EPS and AOM provide halogenated reactive sites for DBP formation, exhibiting diverse aromatic substances and unsaturated (lignin and tannins) compounds. Our results highlight divalent ions acting as a fundamental driving force in DBP formation, suggesting the need for cautious monitoring of divalent ions in karst water., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Currently, Advanced Reduction Process (ARP) is gaining popularity as an alternative to Advanced Oxidation Process (AOP). Though UV/Sulfite process is effective in degrading organic compounds, no investigation has been done using ARP to improve sludge dewaterability. Here, effect of two different ARP's (UV/Sulfite; UV/Sulfide) that generates hydrated electron (e aq - ) and hydrogen atom (H•) in enhancing sludge dewatering was explored. Based on findings, alkaline pH was favourable for ARP's to improve sludge dewatering. At optimal conditions, CST value was 18 and 23.3 s for UV/Sulfite and UV/Sulfide with raw sludge exhibiting 100 s at pH-7.7 respectively. The mechanism revealed that e aq - was the dominant reducing radical along with H•, based on EPR spectra and quenching experiments. UV-Vis, Gaussian fitting, 3D EEM Fluorescence and Synchronous Fluorescence Spectroscopy exhibited higher release of organic matter and aromaticity which agrees with FTIR analysis. Emission peak around 330-380 nm in all samples exhibited Microbial Soluble Products and aromatic protein II., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Current strategies primarily utilize heterotrophic or mixotrophic bioreduction for the simultaneous removal of Cr(VI) and NO 3 - from groundwater. However, given the oligotrophic nature of groundwater, autotrophic bioreduction could be more appropriate, though it remains notably underdeveloped. Here, an autotrophic bioreduction technology utilizing biochar (BC)-assisted zero valent iron (ZVI) is proposed. The pyrolysis temperature of BC was optimized to enhance electron transfer efficiency and reduce extracellular polymeric substances (EPS) accumulation. BC500, with the superior electron transfer capabilities, was the most effective. After an 11-week period, the ZVI + BC500 biotic column still achieved 100% removal efficiency for Cr(VI) and 93.37 ± 0.33% for NO 3 - , with initial concentrations of 26 mg/L and 50 mg/L, respectively. Its performance significantly surpasses that of ZVI alone, effectively reducing the interference of Cr(VI) on denitrification. The presence of quinone and phenolic compounds in BC500, serving as electron-accepting and electron-donating groups, improves the efficiency of electron transfer between ZVI and microbes. Metagenomic analysis showed an increase in the growth of autotrophic bacteria such as Hydrogenophaga spp. and Rhodanobacter denitrificans, and heterotrophic bacteria including Arenimonas daejeonensis and Chryseobacterium shandongense. The promotion facilitates the expression of genes associated with Cr(VI) reduction (chrR, nemA) and denitrification (narG, nirS). BC500 also enhanced EPS production, which facilitates the adsorption and reduction of Cr(VI), mitigating its inhibitory effects on denitrification. Notably, in the ZVI + BC500 biotic column, the accumulated EPS primarily consists of loosely bound EPS rather than tightly bound EPS, potentially reducing the risk of pore clogging during in-situ groundwater treatment., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

The control of organic micropollutants (OMPs) in water environments have received significant attention. Denitrification was reported to exhibit good efficiency to remove OMPs, and the mechanisms involved in are too intricate to be well illustrated. In this study, we selected nitrobenzene [NB] and bisphenol A [BPA] as model pollutants and aimed to unravel the mechanisms of Paracoccus Denitrificans in the removal of OMPs, with a specific emphasis on aerobic behavior during denitrification processes. We demonstrated the formation of extracellular superoxide radicals, i.e., extracellular • O 2 - , using a chemiluminescence probe and found that extracellular polymeric substance adsorption, extracellular • O 2 - , and microbial assimilation contributed approximately 40 %, 10 %, and 50 % to OMPs removal, respectively. Transcriptome analysis further revealed the high expression and enrichment of several pathways, such as drug metabolism-other enzymes, of which a typical aerobic enzyme of polyphenol oxidase [PPO] participates in the degradation of NB and BPA. Importantly, all the immediate products showed a significant decrease in toxicity during the aerobic activity-related OMPs degradation process based on the proposed degradation pathways. This study demonstrates the formation of extracellular • O 2 - and the mechanisms of extracellular • O 2 - - and PPO-mediated OMPs biodegradation, and offers new insights into OMPs control in widely-used denitrification treatment processes., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier Ltd.)

An interesting and potential property of extracellular polymeric substances (EPS) is the hydrogel formation with calcium ions. Aiming at understanding the significant difference in the hydrogel formed between EPS from flocculent and granular sludge, a targeted investigation of the lipopolysaccharides (LPS), one of the important EPS components, was performed. LPS was isolated from the EPS of flocculent and granular sludge, and both the glycan and the lipid A parts of LPS were characterized and compared. The morphology of LPS-calcium (LPS-Ca) aggregates were visualized by the polymyxin B-based fluorescent probe. The LPS constituted about 25 % and 15 % of the EPS from flocculent and granular sludge, respectively. The flocculent sludge LPS showed a lower amount of glycans, shorter glycan chain length, lower molecular weight, and higher possibility of containing unsaturated lipids than the granular sludge EPS. The flocculent sludge LPS-Ca aggregates demonstrated invert structures with the water phase in between, contributing to the fluid-like property of the respect EPS-Ca. In contrast, with the remarkably different chemical structure, LPS-Ca aggregates from granular sludge displayed bilaminar multilayered morphology, contributing to the solid, self-standing hydrogel of EPS-Ca., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

Microalgae, such as Parachlorella kessleri, have significant potential for environmental remediation, especially in removing heavy metals like zinc from water. This study investigates how P. kessleri, isolated from a polluted river in Argentina, can remediate zinc. Using atomic force microscopy (AFM), the research examined the interactions between Zn particles and cells grown with different nitrogen sources-nitrate or ammonium. The results showed that cells grown with nitrate produced extracellular polymeric substances (EPS), while those grown with ammonium did not. Raman spectroscopy revealed distinct metabolic responses based on the nitrogen source, with nitrate-grown cells showing altered profiles after zinc exposure. Zinc exposure also changed the surface roughness and nanomechanical properties of the cells, particularly in those producing EPS. AFM force spectroscopy experiments then confirmed strong Zn binding to EPS in nitrate-grown cells, while interactions were weaker in ammonium-grown cells that lacked EPS. Overall, our results elucidate the critical role of EPS in Zn removal by P. kessleri cells and show that Zn remediation is mediated by EPS adsorption. This study underscores the significance of regulating nitrogen sources to stimulate EPS production, offering insights that are essential for subsequent bioremediation applications., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)

As the primary reservoir of heavy metals in nature, soil is highly susceptible to significant co-contamination with Cd-As-Ni. In current study, extracellular polymeric substances (EPS) from Bacillus subtilis were utilized as a novel improver to simultaneously enhance soil property and restrain ecotoxicity in Cd-As-Ni co-contaminated soil. Our findings revealed that EPS effectively bound and immobilized free Cd, As, and Ni in soil and decreased 49.73 % of soil available Cd, 79.16 % of As and 77.87 % of Ni contents by increasing soil pH, soil organic matter and cation exchange capacity. The EPS was also found to inhibit the Cd-As-Ni induced ecotoxicity in Caenorhabditis elegans by increasing the activities of antioxidant enzymes including superoxide dismutase, glutathione, and catalase. The remediation of EPS showed progressive improvement over time, and maintained a lasting effect after achieving peak efficiency. Our results might provide a new perspective on the potential of EPS in remediation of soil heavy metal pollution and the development and utilization of microbial biomass resources in a wider range., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Microbial biomineralization significantly affects the uranium (U) behavior in the environment. However, the mechanism of microbial biomineralization of U is still not fully understood. In this study, a dominant bacterium (Pseudomonas sp. WG2-6) was isolated from U tail mining area. Abiotic precipitation tests demonstrated that U biomineralization was entirely attributed to the mediation of Pseudomonas sp. WG2-6 when the concentration ratio of exogenous β-glycerophosphate (SGP) to U was 10:1. Pseudomonas sp. WG2-6 exhibited strong immobilization ability towards U (97.59 %) according to batch experiments, and acylamide, carbonyls, and phosphate groups were the main functional groups that interacted with U. Besides, U mainly existed in the form of amorphous U-P complexes after biomineralization by Pseudomonas WG2-6, which could be converted into crystalline nano-minerals H 2 (UO 2 ) 2 (PO 4 ) 2 ·8H 2 O in the presence of SGP. In particular, the formation and structural composition changes of extracellular polymeric substance (EPS) as well as the decrease in U4f binding energy were observed during the U biomineralization process of Pseudomonas sp. WG2-6 in the presence of SGP, indicating that EPS provided the nucleation site for the formation of stable biomineralized products. This work provides new insight into the mechanism of U microbial biomineralization and a theoretical basis for the remediation of U contaminated environments through microbial biomineralization., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier B.V.)

This paper investigates the effects of phenolic-formaldehyde resin microplastics (PF-MPs) with different particle sizes on anaerobic granular sludge (AnGS) and reveals the complex interaction mechanisms between extracellular polymeric substances (EPS) and PF-MPs through the combination of molecular dynamics simulations and spectroscopy. PF-MPs provide a new ecological niche for microorganisms. Microorganisms and EPS can adhere and accumulate on the surface of PF-MPs, producing highly active floc sludge inside the reactor, thereby increasing the chemical oxygen demand (COD) removal rate and methane production of the reactor. However, the high metabolic activity of floc sludge consumes the biodegradable components in EPS, resulting in loose rupture of the sludge particles and reduced particle size. In addition, small particle size S-PF can adhere to the sludge surface，which caused mass transfer barriers and reduced the expression of genes and enzyme activities for the sludge acidification process and the main methanogenic processes. Insufficient internal nutrients lead to endogenous metabolism within the granules, causing internal hollowing, which affects the density and settling performance of the sludge. Monolayer physical adsorption plays a major role in the adsorption of EPS on PF-MPs. 2D-COS and FTIR spectroscopy were used to elucidate the preferential binding of polysaccharides to PF-MPs. This paper explores the fate of PF-MPs in anaerobic systems and demonstrates the important role of EPS in the capture of microplastics by granular sludge, providing a theoretical basis for understanding the migration of microplastics in wastewater treatment., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Extracellular polymeric substances (EPS) are tightly related to the horizontal gene transfer (HGT) of antibiotic resistance genes (ARGs), but often neglected in soil. In this study, nanoscale zero-valent iron (nZVI) was utilized for attenuation of ARGs in contaminated soil, with an emphasis on its effects on EPS secretion and HGT. Results showed during soil microbe cultivation exposed to tetracycline, more EPS was secreted and significant increase of tet was observed due to facilitated HGT. Notably, copies of EPS-tet accounted for 71.39 % of the total tet, implying vital effects of EPS on ARGs proliferation. When co-exposed to nZVI, EPS secretion was decreased by 38.36-71.46 %, for that nZVI could alleviate the microbial oxidative stress exerted by tetracycline resulting in downregulation of genes expression related to the c-di-GMP signaling system. Meanwhile, the abundance of EPS-tet was obviously reduced from 7.04 to 5.12-6.47 log unit, directly causing decrease of total tet from 7.19 to 5.68-6.69 log unit. For the reduced tet, it was mainly due to decreased EPS secretion induced by nZVI resulting in inhibition of HGT especially transformation of the EPS-tet. This work gives an inspiration for attenuation of ARGs dissemination in soil through an EPS regulation strategy., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

The pathway of reducing U(VI) to insoluble U(IV) using electroactive bacteria has become an effective and promising approach to address uranium-contaminated water caused by human activities. However, knowledge regarding the roles of extracellular polymeric substances (EPS) in the uranium reduction process involving in extracellular electron transfer (EET) mechanisms is limited. Here, this study isolated a novel U(VI)-reducing strain, Desulfovibrio vulgaris UR1, with a high uranium removal capacity of 2.75 mM/(g dry cell). Based on a reliable EPS extraction method (45 °C heating), manipulation of EPS in D. vulgaris UR1 suspensions (removal or addition of EPS) highlighted its critical role in facilitating uranium reduction efficiency. On the second day, U(VI) removal rates varied significantly across systems with different EPS contents: 60.8% in the EPS-added system, 48.5% in the pristine system, and 22.2% in the EPS-removed system. Characterization of biogenic solids confirmed the reduction of U(VI) by D. vulgaris UR1, and the main products were uraninite and UO 2 (2.88-4.32 nm in diameter). As EPS formed a permeable barrier, these nanoparticles were primarily immobilized within the EPS in EPS-retained/EPS-added cells, and within the periplasm in EPS-removed cells. Multiple electroactive substances, such as tyrosine/tryptophan aromatic compounds, flavins, and quinone-like substances, were identified in EPS, which might be the reason for enhancement of uranium reduction via providing more electron shuttles. Furthermore, proteomics revealed that a large number of proteins in EPS were enriched in the subcategories of catalytic activity and electron transfer activity. Among these, iron-sulfur proteins, such as hydroxylamine reductase (P31101), pyruvate: ferredoxin oxidoreductase (A0A0H3A501), and sulfite reductase (P45574), played the most critical role in regulating EET in D. vulgaris UR1. This work highlighted the importance of EPS in the uranium reduction by D. vulgaris UR1, indicating that EPS functioned as both a reducing agent and a permeation barrier for access to heavy metal uranium., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Inc. All rights reserved.)

The secretion, accumulation, and composition of extracellular polymeric substances (EPS) are pivotal factors influencing microalgal growth as well as wastewater recycling. Until now, the accumulation and inhibition mechanism of EPS of Chlorella vulgaris during cycling cultivation is not fully understood. The purpose of this study was to explore how different light qualities regulate the secretion, chemical composition, and structure of microalgal EPS, and subsequently influence the recycling of culture wastewater. After four cycles of cultivation, C. vulgaris under green light produced the highest EPS production and lowest biomass production, which were 82% higher and 17% lower, respectively, compared to white light, which yielded the least EPS production and the highest biomass production. EPS under different light qualities all exhibited a fibrillar structure with a sheet-like surface, but differed in composition. Compared with the other groups, EPS under green light showed a significant increase in polysaccharides, proteins, and humic acid-like compounds, as well as an increased proportion of arabinose and rhamnose, according to monosaccharide composition analysis. Transcriptome analysis indicated that the up-regulation of metabolic pathways linked to glycolysis/gluconeogenesis, TCA cycle, lipid synthesis, and ABC transporters promoted EPS accumulation. Additionally, EPS could target light-harvesting complex (LHC) and electron transport chain, down-regulating the photosynthetic pathway, which ultimately inhibited microalgal growth under green light. This study provides a theoretical foundation for the light regulation and circulation culture of microalgae, as well as for microalgal wastewater treatment., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Although biochar (BC)-assisted microbial electrolysis cells (MEC) has been shown to improve anaerobic digestion (AD) performance of waste activated sludge (WAS), the underlying mechanisms remain unclear. This study conducted an in-depth investigation into the mechanism based on analyses of extracellular polymeric substances (EPS) characteristics, enzyme activities and multi-omics. The results showed that compared with the control group, methane production improved by 16.73 %, 21.32 %, and 29.37 % in the BC, MEC, and BC-assisted MEC (BC-MEC) groups, respectively. The reconfiguration of the protein secondary structure increased the hydrophobicity of the EPS, thereby promoting microbial aggregation. In addition, partial least-squares path modeling (PLS-PM) and mantel test based on the enzyme activity and multi-omics analyses revealed that the promotional effect of MEC on the hydrolysis of WAS was superior to that of BC, while BC was more advantageous in promoting electron transfer and biofilm formation regulated by quorum sensing. The synergistic effects of BC and MEC were exemplified in the BC-MEC group. g&#95;norank&#95;Aminicenantales responsible for the hydrolysis of WAS was enriched (29.6 %), and the activities of hydrolytic enzymes including α-glucosidases and proteases were increased by 29.1 % and 43.6 %, respectively. Further, the expressions of genes related to acyl homoserine lactones (AHLs) and diffusible signal factor (DSF) in quorum sensing systems, as well as the genes related to hydrogenase involved in electron transfer (mbhJKL, hyfB-JR, hypA-F, and hoxFHUY), were up-regulated in the BC-MEC group. This facilitated electron transfer and microbial communication, consequently enhancing methane production. This research significantly advances the understanding of the mechanism by which BC-assisted MEC enhances AD performance and provides valuable insights into strategies for improving energy recovery from WAS., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

The ever-increasing plastic waste accumulation in the marine environment necessitates a deeper understanding of microalgae interactions with micro- and nanoplastics (MNP), and the role of extracellular polymeric substances (EPS). EPS, known for its adhesive properties and produced as an algal stress response, may facilitate aggregation of both algae and MNPs, thereby impacting ecological and hydrodynamic processes such as the trophic transfer or vertical transport of MNPs. Moreover, gaining a deeper understanding of the impact of weathering processes on the potential toxicological effects of plastic particles, and the comparative significance of plastic-specific effects relative to those of naturally occurring particles such as kaolin clay, is imperative. Therefore, this study investigated the impact of fragmented, polydisperse virgin polyethylene terephthalate (PET, Daverage = 910 nm) and weathered PET (Daverage = 1700 nm) on the growth and the production of EPS of Rhodomonas salina. Algae were exposed to a range of low MNP concentrations (10, 100 and 1000 and 10,000 MNPs ml-1) for 11 days. A natural particle control (kaolin, Daverage = 1600 nm) was deployed to differentiate particle effects from plastic effects. It was observed that exposure to both weathered PET and virgin PET resulted in initially increased growth rates (7.80 % and 7.28 % respectively), followed by significant decreases in algae cell density (-30.1 % and -11.2 % respectively). Furthermore, exposure to weathered PET caused a simultaneous elevation in cellular EPS production (76.51 %). The effects of plastics were significantly larger than the effect of kaolin. Also, the observed effects were amplified by the weathering of the plastics. These observations underscore the interactions between particle type, age and concentration, and their distinct impacts on algae density and growth inhibition. The observations indicate a role for EPS as an algal protection mechanism, potentially affecting the environmental fate of MNP - microalgae aggregates., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Marie Sioen reports financial support was provided by Research Foundation Flanders. Maaike Vercauteren reports financial support was provided by Special Research Fund of Ghent University. The other authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Highly hydrophilic extracellular polymeric substance (EPS) with gel-like structure seriously plagues the development of sludge deep dewatering. Oxysulfur radicals-based oxidation driven by iron-bearing mineral proposes a promising strategy for effective EPS decomposition. However, the transformation and involved interaction mechanisms of aromatic proteins are still controversial due to the complex EPS structure. Herein, sulfite mediated siderite (denoted as Fe(II)/S(IV)) was developed for targeted transformation aromatic amino acids in EPS oxidation to strengthen sludge solid-liquid separation. The enhanced sludge dewaterability were benefited from the Fe(II)/S(IV) bonded interaction assisted by Fe 3+ /Fe 2+ as redox interface that facilitating the release of intracellular bound water via diminish the hydrophily and bind strength with solid protons. The amide region nitrogen of aromatic amino acids (especially tyrosine and tryptophan) originating from EPS presented looser structure and lower spatial site resistance, which were attributed to the exposure of hydrophobic sites in amino groups after Fe(II)/S(IV) treatment. Furthermore, the effective decline of aromatic amino acids in inner layer-EPS (loosely bound EPS and tightly bound EPS) was directed from Fe-N targeted interaction by triggering a series of sulfate-based radical chain reactions. The good correlation between electron transfer amount (R 2 = 0.926) and Fe-N (R 2 = 0.925) with bonding interaction demonstrated that the complexation of aromatic amino acids with Fe sites on siderite/sulfite via Fe-N bonds, accounting for efficient sludge solid-liquid separation. This study deepens the understanding of sludge organic matter targeted transformation and provides a tactic for iron-based conditioning of sludge., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Structural extracellular polymeric substances (SEPS) as valuable biopolymers, can be extracted from waste activated sludge (WAS). However, the extraction yield is typically low, and detailed information on SEPS characterizations, as well as proper treatment of the sludge after SEPS extraction, remains limited. This study aimed to optimize the conditions of heating-Na 2 CO 3 extraction process to increase the yield of SEPS extracted from WAS. Subsequently, SEPS were characterized, and, for the first time, insights into their protein composition were uncovered by using proteomics. A maximum SEPS yield of 209 mg g -1 volatile solid (VS) was obtained under optimal conditions: temperature of 90 °C, heating time of 60 min, Na + dosage of 8.0 mmol/g VS, and pH required to precipitation of 4.0, which was comparable to that from the aerobic granular sludge reported in literature. Proteomics analysis unveiled that the proteins in SEPS primarily originated from microorganisms involved in nitrogen fixation and organic matter degradation, including their intracellular and membrane-associated regions. These proteins exhibited various catalytic activities and played crucial roles in aggregation processes. Besides, the process of SEPS extraction significantly enhanced volatile fatty acid (VFA) production during the anaerobic fermentation of residual WAS after SEPS extraction. A maximum VFA yield of 420 ± 14 mg COD/g VS added was observed in anaerobic fermentation of 10 d, which was 77.2 ± 0.1 % higher than that from raw sludge. Mechanism analysis revealed that SEPS extraction not only improved WAS disintegration and solubilization but also reduced the relative activity of methanogens during anaerobic fermentation. Moreover, SEPS extraction shifted the microbial population during anaerobic fermentation in the direction towards hydrolysis and acidification such as Fermentimonas sp. and Soehngenia sp. This study proposed a novel strategy based on SEPS extraction and VFA production for sludge treatment, offering potential benefits for resource recovery and improved process efficiency., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

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., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Background: Biofilm, a common drug-resistant phenotype of Staphylococcus aureus (S. aureus), demonstrates significant drug resistance and recurrence due to its extracellular polymeric substance (EPS) barrier and subsequent bacterial migration. Hence, there is an urgent need for effective solutions to mitigate the hazards posed by biofilms., Result: This study developed a stable, low-toxicity multifunctional nanomicelle, GLA@SOL/EYL, by encapsulating glabridin (GLA) using sophorolipid (SOL) and egg yolk lecithin (EYL). Optimizations were performed for the hydration medium, the ratio of carrier materials to GLA, and EYL additions. GLA@SOL/EYL exhibited a particle size of 122.1 ± 0.8 nm and a surface potential of -66.4 ± 1.7 mV, endowing it with the ability to permeate biofilms EPS effectively. GLA@SOL/EYL encapsulated 98.3 ± 1.2 % of GLA and demonstrated a slow-release effect, significantly enhancing the bioavailability of GLA. The addition of EYL reduced the hemolytic toxicity of GLA@SOL/EYL and improved its encapsulation rate and stability. GLA@SOL/EYL reduced the minimum inhibitory concentration of GLA to 8 μg/mL and extended its inhibitory effect at low concentrations by rapidly disrupting the structural integrity of S. aureus. GLA@SOL/EYL may penetrate biofilms to disperse EPS and remove twice as much biofilm as GLA alone, thereby eliminating 99.99 % of S. aureus within biofilms, compared to 99 % bactericidal efficacy of GLA. Additionally, GLA@SOL/EYL inhibited 63.8 ± 1.8 % of biofilm formation by affecting the expression of the srtA gene, thereby reducing the expression of cell wall-anchoring protein genes. In contrast, the biofilm inhibition rates of GLA and blank micelles were less than 10 %., Conclusion: GLA@SOL/EYL utilizes the nanoparticle effect to penetrate biofilms and deliver antimicrobial GLA. The SOL disperses the biofilm matrix while GLA is released to kill S. aureus, preventing bacterial dissemination and colonization. Thus, GLA@SOL/EYL presents an innovative strategy for effectively eradicating S. aureus biofilms and preventing new hazards in a one-step approach., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier Inc.)

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., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Tire wear particles (TWP) represent a distinctive form of microplastics (MPs) that are widely distributed in aquatic ecosystems. However, the toxicity of various types of TWP on phytoplankton remain to be further explored. Thus, three different TWPs originating from replaced bicycle, car, and electro-mobile tire (marked as BTWP, CTWP, and ETWP) were selected and their long-term biological influences on Chlorella vulgaris were investigated. Results demonstrated TWPs showed a concentration-dependent growth promotion of Chlorella vulgaris, with a maximum promotion rate reached to 40.51 % (10 mg/L, 10 d), 23.5 % (80 mg/L, 12 d), and 28.7 % (20 mg/L, 12 d) in the presence of BTWP, CTWP and ETWP, respectively. Meanwhile, TWPs could stimulate the secretion of EPS and induce oxidative stress. EPS analysis revealed the increase of polysaccharides could protect the cell from the direct contact with TWP particles. Moreover, the increased concentration of EPS also helps to induce the settlement of TWP and reduce the leachate release. The release of TWP into the environment could act as an accelerator for the growth of Chlorella vulgaris, which might further change the normal physicochemical behaviors of algae colony in aquatic system. Our findings provide new insights into the toxicity mechanism of TWPs on freshwater algae and valuable data on environmental risk assessment of TWPs., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

As an important potential dust suppression method, the slow onset time is one of the key factors that restrict the effect of microbial dust suppressant. In the early stage, we have confirmed that extracellular polymeric substances (EPS) can improve the dust suppression effect by wetting coal dust and increasing Ca 2+ nucleation sites. Therefore, in this study, chitosan (CTS) and bovine serum albumin (BSA) in different ratios (CTS: BSA = 1:1, 1:2, 2:1) as model molecules of EPS were combined with Bacillus subtilis to prepare efficient and fast microbial dust suppressants. Furthermore, the interaction forces were analyzed through molecular dynamics simulation. Results showed that adding CTS and BSA would improve the dust suppression effect, and the dust suppression effect was the best when the ratio of CTS: BSA was 1:2. In addition, the contact angle decreased as the BSA content increased. The Fourier transform infrared spectroscopy (FTIR) results showed that when the ratio of CTS to BSA was 1:2, the dust suppressants were easier to interact with coal dust by the key functional groups and form calcite type CaCO 3 . The molecular dynamics simulation results showed that the main interaction was Van der Waals force. In addition, the interaction force was strongest when CTS: BSA was 1:2, increasing by 137% compared with the microbial dust suppressants without CTS or BSA. This study provides theoretical support for the development of efficient and rapid microbial dust suppressants., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Extracellular polymeric substances (EPS), which envelop on fungal hyphae surface, interact strongly with minerals and play a crucial role in the formation of nanoscale minerals during biomineralization in nature environments. However, it remains poorly understood about the molecular mechanisms of nanominerals (i.e., iron nanominerals) formation in fungal EPS halos during fungus-mineral interactions. This process is vital because fungi typically grow attached to various mineral surfaces in nature. According to the changes of thickness of the fungal cell and EPS layers during the Trichoderma guizhouense NJAU 4742 and hematite cultivation experiments, we found that fungal biomineralization could trigger the formation of EPS layers. Fe-dominated nanominerals, aromatic C (283-286.1 eV), alkyl C (287.6-288.3 eV), and carboxylic C (288.4-289.1 eV) were the dominant chemical groups on the EPS layers, as determined by nanoscale secondary ion mass spectrometry (NanoSIMS), high-resolution transmission electron microscope (HRTEM), and carbon 1s near-edge X-ray absorption fine structure (NEXAFS) spectroscopy. Further, evidence from Fe K-edge X-ray absorption near-edge structure (XANES) and X-ray photoelectron spectroscopy (XPS) spectra indicated that oxygen vacancy (O V ) was formed on the Fe-dominated nanomineral surface during fungus-mineral interactions, which played an important role in catalyzing H 2 O 2 decomposition and HO∗ production. Taken together, the intrinsic peroxidase-like activity by reactive oxygen species (ROS) could modulate the Fe-dominated nanominerals formation in EPS layers to newly form a physical barrier between the cell and the external environments around hyphae, providing novel insights into the effects of ROS-mediated fungal-mineral interactions on fungal nutrient recycling, attenuation of contaminants, and biological control in nature environments., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier Ltd.)

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., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Nitrogen biotransformation plays a vital role in the metabolism of microbial communities in sewers. Extracellular polymeric substances (EPS) secreted by microbial communities can form gel-like sewer sediments, causing clogging of the sewer. However, knowledge on the effects of varying nitrogen conditions on the erosion resistance of sewer sediments and EPS produced by sewer microorganisms is limited. In this study, two typical organic/inorganic nitrogen ratios of sewage were reproduced in simulated sewer reactors, i.e., 3/7 (R1 group) and 7/3 (R2 group). Higher organic nitrogen (ON) concentrations were found to increase the critical erosion shear stress by 26.43%; this was ascribed to increased particle diameter, weakened electrostatic repulsion of sediments and stimulated EPS secretion in the R2 group. The protein and polysaccharide contents of the R2 group were 48.84% and 34.25% higher than those of the R1 group, respectively, which was supported by increased gene abundances for aromatic amino acid synthesis, general secretory pathways of protein, and synthesis of precursors and polysaccharides. Tightly-bound EPS in R2 group exhibited increased contents of hydrophobic protein secondary structures and intermolecular hydrogen bonds, thereby promoting the formation of gel-like sediment structures with enhanced erosion resistance. However, the microbial diversity and the abundance of key genes involved in EPS generation and secretion (e.g., tyrB, yajC, secB, gumF, and gumH) obviously decreased in the R1 group. Moreover, high ON concentrations increased microbial diversity and enhanced microbial glycolysis, tricarboxylic acid cycle and ammonium assimilation. This study reveals the formation mechanisms of EPS in sewer sediments under different nitrogen conditions and their effects on sediment erosion resistance, which contributed to improved sewer system operation and sewer sediment control., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Extracellular polymeric substances (EPS) have demonstrated significant benefits for reducing multivalent metal contamination. Using Achromobacter xylosoxidans BP1 isolated from a coal chemical site in China, this study elucidated the contribution of EPS production to Cr (VI) reduction and revealed its biological removal mechanism. BP1 grew at an optimum pH of 8 and the lowest inhibitory concentration of Cr(VI) was 300 mg/L. The spent medium completely removed Cr(VI), whereas resting cells were only able to remove 10.47 % and inactivated cells were nearly incapable of Cr(VI) removal. S-EPS and B-EPS reduced Cr(VI) by 98.59 % and 11.64 %, respectively. SEM-EDS analysis showed that the BP1 cells were stimulated to produce EPS under Cr stress. The XPS results showed that 29.63 % of Cr(VI) was enriched by intracellular bioaccumulation or biosorption and 70.37 % of Cr(VI) was reduced by extracellular enzymes to produce Cr(OH) 3 and organic Cr(III) complexes. According to FTIR, EPS with -OH, COO-, and amide groups supplied binding sites and electrons for the reductive adsorption of Cr(VI). Genomic studies showed that BP1 primarily produces extracellular polysaccharides, metabolises sulphur and nitrogen, and reduces reactive oxygen species damage as a result of DNA repair proteases., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Gut digestion by earthworms (GDE) is a crucial step in vermicomposting, affecting the fate of antibiotic resistance genes (ARGs) in vermicompost sludge. The extracellular polymeric substance (EPS) matrix of sludge is an important space for ARG transfer. However, the effect of GDE on EPS-associated ARGs remains unclear. Therefore, this study explored the role of GDE in driving the transfer of ARGs within different EPS layers in sludge. For this, the changes in intracellular ARGs and EPS-associated ARGs in sludge were analyzed after 5 days of the GDE process. The results showed that after the GDE process, both nitrate and dissolved organic carbon significantly increased in all EPS layers of sludge, while the proteins and polysaccharides only enhanced in soluble and loosely bound EPS of sludge. In addition, a 7.0% decrease in bacterial diversity was recorded after the GDE process, with a functional bacterial community structure emerging. Moreover, the absolute abundance of total ARGs and mobile genetic elements decreased by 90.71% and 61.83%, respectively, after the GDE process. Intracellular ARGs decreased by 92.1%, while EPS-associated ARGs increased by 4.9%, indicative of intracellular ARG translocation into the EPS during the GDE process. Notably, the ARGs exhibited significant enrichment in both the soluble and loosely bound EPS, whereas they were reduced in the tightly bound EPS. The structural equation modeling revealed that the GDE process effectively mitigated the ARG dissemination risk by modulating both the EPS structure and microenvironment, with the organic structure representing a primary factor influencing ARGs in the EPS., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Inc. All rights reserved.)

The interaction between extracellular polymeric substances (EPS) in municipal sludge and antibiotics in wastewater is critical in wastewater treatment, resource recovery, and sludge management. Therefore, it is increasingly urgent to investigate the distribution coefficient (Log K) of sulfonamide antibiotics (SAs) in EPS, particularly in sludge-derived dissolved organic carbon (DOC) and aqueous phase systems. Herein, through balance experiments, the concentrations of SAs were determined using alkaline extraction EPS (AEPS) and alginate-like extracellular polymer (ALE) systems, and the Log K DOC values were determined. The results showed that the Log K DOC of AEPS was higher than that of ALE, which exhibited a negative K DOC value, indicating an inhibitory effect on dissolution. For the three SAs studied, the Log K DOC values were in the following order: sulfamethoxazole > sulfapyridine > sulfadiazine. This order can be attributed to the differing physicochemical properties, such as polarity, of the SAs. Three-dimensional excitation-emission matrix fluorescence spectra and fitting results indicated a lack of aromatic proteins dominated by tryptophan and humus-like substances in ALE. Meanwhile, the hydrophobic interaction of aromatic proteins dominated by tryptophan was the main driving force in the binding process between AEPS and SAs., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier Inc.)

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., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

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., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Ltd.. All rights reserved.)

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., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Although the antimicrobial mechanisms of nanomaterials have been extensively investigated, bacterial defense mechanisms associated with AgNPs have not been fully elucidated. We here report that dissolved Ag + (>0.05 μg mL -1 ) displayed higher toxicity on cell growth of strain Cupriavidus metallidurans BS1 (GCA&#95;003260185.2) in comparison to 2 and 20 nm AgNPs. The genes necessary for synthesis of distinct abundance and composition of extracellular polymeric substances (EPS) were induced in strain BS1 exposed to Ag stress. This resulted in 20.1% (Ag(I)-EPS) and 24.2% (2 nm AgNPs-EPS) of the CO band integrated intensities being converted into C-OH/C-O-C group vibrations and the Ag-O bond was formed between EPS and 20 nm AgNPs. Meanwhile, the expression of primary resistance genes of the cus, sil and cup operon encoding HME-RND-driven efflux systems as well as a P IB1 -type ATPase (CupA) were significantly induced after exposure to Ag(I), 2 and 20 nm AgNPs, respectively. Furthermore, distinct genes involved in biosynthesis pathways responsible for production of EPS were induced to relieve the toxicity of Ag(I), 2 nm and 20 nm AgNPs. This combined action is one potential reason why strain BS1 displayed distinct resistances in response to Ag(I) compared to 2 and 20 nm AgNPs. This work will help in understanding processes important in bacterial defensive mechanisms to AgNPs., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier Ltd.)

The transport and retention of bacteria in porous media, such as aquifer, are governed by the solid-liquid interface characteristics and bacterial mobility. The secretion of extracellular polymeric substance (EPS) by bacteria modifies their surface property, and thereby has effects on their adhesion to surface. The role of EPS in bacterial mobility within saturated quartz sand media is uncertain, as both promoting and inhibitory effects have been reported, and underlying mechanisms remain unclear. In this study, the effects of EPS on bacterial transport behavior and possible underlying mechanism were investigated at 4 concentrations (0 mg L -1 , 50 mg L -1 , 200 mg L -1 and 1000 mg L -1 ) using laboratory simulation experiments in conjunction with Extend Derjaguin-Landau-Verweu-Overbeek (XDLVO) modeling. The results showed that EPS facilitated bacterial mobility at all tested concentrations. It could be partially explained by the increased energy barrier between bacterial cells and quartz sand surface in the presence of EPS. The XDLVO sphere-plate model predicted that EPS induced a higher electrostatic double layer (EDL) repulsive force, Lewis acid-base (AB) and steric stabilization (ST), as well as a lower Lifshitz-van der Waals (LW) attractive force. However, at the highest EPS concentration (1000 mg L -1 ), the promotion of EPS on bacterial mobility weakened as a result of lower repulsive interactions between cells, which was supported by observed enhanced bacterial aggregation. Consequently, the increased aggregation led to greater bio-colloidal straining and ripening in the sand column, weakening the positive impact of EPS on bacterial transport. These findings suggested that EPS exhibited concentration-dependent effects on bacterial surface properties and transport behavior and revealed non-intuitive dual effects of EPS on those processes., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Aerobic granular sludge (AGS) is a powerful biotechnological tool capable of treating multiple pollutants simultaneously. However, the granulation process and pollutant removal efficiency still need to be further improved. In this study, Fe 2 O 3 - and MnO 2 -surface-modified straw foam-based AGS (Fe 2 O 3 @SF-AGS and MnO 2 @SF-AGS), with an average particle size of 3 mm, were developed and evaluated. The results showed that surface modification reduced the hydrophobic groups of carriers, facilitating the attachment and proliferation of microorganisms. Notably, MnO 2 @SF-AGS showed excellent granulation performance, reaching a stable state about one week earlier than the unmodified SF-AGS. The polymeric substance content of MnO 2 @SF-AGS was found to be 1.28 times higher than that of the control group. Furthermore, the removal rates for NH 4 + -N, TN, and TP were enhanced by 27.28%, 12.8%, and 32.14%, respectively. The bacterial communities exhibited significant variations in response to different surface modifications of AGS, with genera such as Saprospiraceae, Terrimonas, and Ferruginibacter playing a crucial role in the formation of AGS and the removal of pollutants specifically in MnO 2 @SF-AGS. The charge transfer of metal ions of MnO 2 @SF promotes the granulation process and pollutant removal. These results highlight that MnO 2 @SF-AGS is an effective strategy for improving nitrogen and phosphorus removal efficiency from wastewater., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier Ltd.)

Wastewater treatment technologies opened the door for recovery of extracellular polymeric substances (EPS), presenting novel opportunities for use across diverse industrial sectors. Earlier studies showed that a significant amount of phosphorus (P) is recovered within extracted EPS. P recovered within the extracted EPS is an intrinsic part of the recovered material that potentially influences its properties. Understanding the P speciation in extracted EPS lays the foundation for leveraging the incorporated P in EPS to manipulate its properties and industrial applications. This study evaluated P speciation in EPS extracted from aerobic granular sludge (AGS). A fractionation lab protocol was established to consistently distinguish P species in extracted EPS liquid phase and polymer chains. 31 P nuclear magnetic resonance (NMR) spectroscopy was used as a complementary technique to provide additional information on P speciation and track changes in P species during the EPS extraction process. Findings showed the dominance of organic phosphorus and orthophosphates within EPS, besides other minor fractions. On average, 25% orthophosphates in the polymer liquid phase, 52% organic phosphorus (equal ratio of mono and diesters) covalently bound to the polymer chains, 16% non-apatite inorganic phosphorus (NAIP) precipitates mainly FeP and AlP, and 7% pyrophosphates (6% in the liquid phase and 1% attached to the polymer chains) were identified. Polyphosphates were detected in initial AGS but hydrolyzed to orthophosphates, pyrophosphates, and possibly organic P (forming new esters) during the EPS extraction process. The knowledge created in this study is a step towards the goal of EPS engineering, manipulating P chemistry along the extraction process and enriching certain P species in EPS based on target properties and industrial applications., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

This study employed spectroscopy, metagenomics, and molecular simulation to investigate the inhibitory effects of Cd(II) and Cu(II) on the anammox system, examining both intracellular and extracellular effects. At concentrations of 5 mg/L, Cd(II) and Cu(II) significantly reduced nitrogen removal efficiency by 41.46 % and 62.03 %, respectively. Additionally, elevated metal concentrations were correlated with decreased extracellular polymeric substances (EPS), thereby reducing their capacity to absorb heavy metals, particularly Cu(II), which decreased from 76.47 % to 14.67 %. Spectral analysis revealed alterations in the secondary structures of EPS induced by Cd(II) and Cu(II), decreasing the ratio of extracellular protein α-helix to (β-sheet + random coil), which resulted in looser extracellular protein configurations. The results of the metagenomics study showed that the abundance of Candidatus Kuenenia and its genes encoding nitrogen removal-related enzymes was reduced. The abundance of hzs-γ was reduced by 35.09 % at a concentration of 5 mg/L Cu(II). Conversely, genes associated with metal efflux enzymes, like czcR, increased by 54.86 % at 2 mg/L Cd(II). Molecular docking revealed robust bindings of Cd(II) to HZS-α (-342.299 ± 218.165 kJ/mol) and Cu(II) to HZS-γ (-880.934 ± 55.526 kJ/mol). This study elucidated the inhibitory mechanisms of Cd(II) and Cu(II) on the anammox system, providing insights into the resistance of anammox bacteria to heavy metals., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Dissemination of antibiotic resistance genes (ARGs) has become a critical threat to public health. Activated sludge, rich in extracellular polymeric substances (EPS), is an important pool of ARGs. In this study, mechanisms of conjugation transfer of ARGs induced by EPS, including tightly bound EPS (TBEPS), soluble EPS (SEPS), and loosely bound EPS (LBEPS), were explored in terms of molecular diversities and electron transfer properties of EPS. Conjugation transfer frequency was increased by 9.98-folds (SEPS), 4.21-folds (LBEPS), and 15.75-folds (TBEPS) versus the control, respectively. Conjugation-related core genes involving SOS responses (9 genes), membrane permeability (18 genes), intercellular contact (17 genes), and energy metabolism pathways (13 genes) were all upregulated, especially in the presence of TBEPS. Carbohydrates and aliphatic substances in SEPS and LBEPS were contributors to ARG transfer, via influencing reactive oxygen species (ROS) formation (SEPS) and ROS and adenosine triphosphate (ATP) production (LBEPS). TBEPS had the highest redox potential and greatest lability and facilitated electron transfer and alternated respiration between cells, thus promoting ARG transfer by producing ATP. Generally, the chemical molecular characteristics and redox properties of EPS facilitated ARG transfer mainly by influencing lipid peroxidation and ATP, respectively., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Extracellular polymeric substances (EPS) secreted by organisms tend to encapsulate microplastics (MPs), forming an EPS-corona that affects the fate of MPs in marine ecosystems. However, the impact of the EPS-corona on the biotoxicity of MPs to marine organisms remains poorly understood. Herein, the effect of the EPS-corona on the toxicity of polystyrene (PS) MPs of different sizes (0.1 and 1 µm) to Skeletonema costatum (S. costatum) was investigated. The preferential adsorption of medium molecule weight (∼55 kDa) proteins onto PS MPs mainly contributed to the EPS-corona formation, decreasing the surface charge negativity of small-sized PS MPs (0.1 µm) by 72.4 %. Nitrogen (N) and oxygen (O) moieties in polysaccharides and proteins were identified as the preferential adsorption sites in the EPS-PS MPs interaction. Density functional theory (DFT) calculations confirmed the nuclear magnetic resonance spectroscopy (NMR) results, revealing that the binding mode between EPS and PS MPs was mainly hydrogen bonding. In addition, EPS-corona increased the cell density of S. costatum by 35.5-36.0 % when exposed to small-sized PS MPs (0.1 µm, 25-50 mg/L). These findings provide new insights into how EPS-corona affects the environmental fate and ecological risks associated with micro- and nano-sized plastics in marine ecosystems., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

This experiment aimed to study the effects of adding the exogenous signaling molecule N-hexanoyl-homoserine lactone (C6-HSL) on the anaerobic digestion of food wastewater at low temperature (15 °C). Daily addition of 0.4 μmol C6-HSL increased the average chemical oxygen demand removal from 45.98% to 94.92%, while intermittent addition (adding 2 μmol C6-HSL every five days) increased it from 45.98% to 72.44%. These two modes of C6-HSL addition increased protease and acetate kinase activity by 47.99%/8.04% and 123.26%/127.91% respectively, and increased coenzyme F 420 concentrations by 15.79% and 63.16%, respectively. The regulation of loosely bound extracellular polymeric substances synthesis was influenced by C6-HSL, which increased protein and polysaccharide content in sludge. The relative abundance of Firmicutes and Bacteroidetes increased following addition of C6-HSL. After continuous addition of C6-HSL, the relative abundance of related functional genes such as amy, apgM, aceE, and accC increased, indicating that methanogens obtained sufficient substrate. The abundance of glycolysis-related functional genes such as glk, pfk, pgi, tpiA, gap, pgk, gpmA, eno, and pyk increased after the addition of C6-HSL, ensuring the efficient transformation and absorption of organic matter by anaerobic sludge at low temperatures. This study provides new comprehensive insights into the mechanism behind the enhancement of food wastewater anaerobic digestion by C6-HSL at low temperature., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

The extremely slow growth rate of anaerobic ammonia oxidation (anammox) bacteria limits full-scale application of anammox process worldwide. In this study, extracellular polymeric substances (EPS)-coated polypropylene (PP) carriers were prepared for biofilm formation. The biomass adhesion rate of EPS-PP carrier was 12 times that of PP carrier, and EPS-PP achieved significant enrichment of E. coli BY63. The 120-day continuous flow experiment showed that the EPS-PP carrier accelerated the formation of anammox biofilm, and the nitrogen removal efficiency increased by 10.5 %. In addition, the abundance of Candidatus Kuenenia in EPS-PP biofilm was 27.1%. Simultaneously, amino acids with high synthesis cost and the metabolites of glycerophospholipids related to biofilm formation on EPS-PP biofilm were significantly up-regulated. Therefore, EPS-PP carriers facilitated the rapid formation of anammox biofilm and promoted the metabolic activity of functional bacteria, which further contributed to the environmental and economic sustainability of anammox process., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Microbial electrolysis cells (MEC) have emerged as a prominent technology for the treatment of antibiotics-containing wastewater in recent years. However, there remains a dearth of comprehensive exploration regarding the influence of extracellular polymers substances (EPS) on the distribution and transmission of antibiotic resistance genes (ARGs) in MEC. In this study, we quantified the distribution of ARGs in MEC by Fluorescence quantitative polymerase chain reaction and explored with emphasis on impact of EPS component on ARGs transmission at under different concentrations of roxithromycin. Results showed that the absolute abundance of ARGs in the electrode biofilm was 1-2 orders of magnitude higher than that in the anolyte. Specifically, EPS-associated ARGs accounted for 2.31%-11.18% of ARGs in electrode biofilm. The presence of elevated roxithromycin concentration led to electroactive microorganisms (Geobacter and Geothrix) as potential hosts of ARGs. In addition, both protein and polysaccharide content in the electrode biofilm increased with increasing roxithromycin concentration and showed positive correlations with EPS-associated ARGs. Fluorescence quenching experiments further elucidated that tryptophan and tyrosine residues in EPS could bind to ARGs effectively, contributing the hindering the ARGs transmission between hosts. Therefore, increased EPS content within electrode biofilm could reduce the concentration of ARGs present in anolyte while also influencing ARGs distribution throughout MEC. This study provides valuable insights into the distribution of ARGs in MEC systems and the role of EPS in regulating ARGs migration., Competing Interests: Declaration of competing interest We declare that we have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)