Soluble Extracellular Polymeric Substances Research Articles

Deciphering stratified structure and microbiota assembly of biofilms from a pyrite-based biofilter driven by mixotrophic denitrification

The precise structure and assembly process of pyrite-based biofilms remain poorly understood. The polysaccharides (PN), proteins (PS), and extracellular DNA were enriched in the soluble extracellular polymeric substance (EPS), loosely bound EPS, and tightly bound EPS, respectively, indicating a significant stratified structure of biofilms. The tryptophan facilitated mixotrophic metabolic processes. Both dominant (>1%) and rare species (<0.01 %) harbored core bacteria, including sulfur autotrophic bacteria, sulfate-reducing bacteria, and heterotrophic bacteria. Furthermore, partial least-squares path modeling quantified the contributions of total phosphorus (TP) (λ = 0.32), dissolved organic matter (DOC) (λ = 0.29), and NH4+–N (λ = 0.26) to variations in the microbial community. Nonmetric multidimensional scaling analysis revealed three distinct stages in biofilm development: colonization (0–36 d), succession (36–149 d), and maturation/old (149–215 d). Furthermore, neutral community model indicated that stochastic processes drove the colonization and maturation/old stages, while deterministic processes dominated the succession stage. This study offered valuable insights into the regulation of pyrite-based engineered ecosystems.

Response of nitrifiers to gradually increasing pH conditions in a membrane nitrification bioreactor: Microbial dynamics and alkali-resistant mechanism

Nitrification and nitrifiers are pH-sensitive especially under the alkaline environment in the activated sludge system. However, it is unclear how nitrifiers and nitrification respond to long-term alkaline environment. This study employed a continuous flow membrane nitrification bioreactor to investigate the dynamics of nitrification efficiency and microbial community adaptation under a 320-day alkaline operation. Results showed that activated sludge adapted remarkably to a progressive increase in pH from 7.5 to 10.0, achieving robust nitrification with average ammonia removal efficiencies of 96.6 ± 2.2%. Subsequently, an integrated alkali-resistant mechanism of nitrifiers was proposed. Specifically, under the long-term operation of pH 10.0, certain bacteria secreted enhanced extracellular acidic polysaccharides (i.e., up to 10.95 ± 0.27 mg·g−1 MLVSS in soluble extracellular polymeric substances (EPS)) and acidic organic compounds (e.g., humic acids increased by 1.47-fold in tightly bounded EPS) to neutralize external alkalinity. Moreover, significant enrichments in both the ammonia oxidizing bacteria Nitrosomonas (by 1.3%) and the nitrite oxidizing bacteria Nitrospira (by 5.4%) were observed in a 170-day operation of pH 10.0 condition. Meanwhile, norank_f__JG30-KF-CM45 (2.0%) and Rhodobacter (0.9%) also contributed to ammonia removal at pH 10.0. On the cellular-level, bacteria enabled to maintain intracellular pH stabilization primarily through cation/proton antiporters, evidenced by significant increases in NhaA, TrkA and KefB activities by 98.0%, 151.7% and 115.2%, respectively. A 43.1% increase in carbonic anhydrase activity also facilitated consumption of aqueous OH− ions through biomineralization, leading to CaCO3 deposition on microbial surface. These findings further enhanced understandings of physiological adaptation of nitrifiers in the long-term alkaline activated sludge system.

A comprehensive study on the effects of electrocoagulation integrated in a membrane bioreactor treating sunflower oil refinery wastewater on treatment performance, biological properties, and fouling behavior.

This study evaluated the effects of electrocoagulation integrated in a laboratory-scale membrane bioreactor (MBR), namely EC-MBR, on the treatment performance, activated sludge morphological characterization, and membrane fouling of MBR treating actual sunflower oil refinery wastewater. The EC-MBR system exhibited significantly higher chemical oxygen demand (COD) and oil and grease (O&G) removal efficiency compared to the MBR system. Additionally, both systems achieved excellent turbidity removal, with a percentage above 99%. The membrane fouling rate was higher in the EC-MBR system compared to the MBR system. Despite the decrease in the soluble microbial product (SMP) and extracellular polymeric substance (EPS) concentration in the EC-MBR system, especially their protein fraction, the significant increase in MLSS and carbohydrates/protein ratio, and the decrease in the mixed liquor and the cake layer particles size were the main membrane fouling factors. The membrane fouling resistance distribution showed that the EC-MBR system had a higher percentage of pore blocking resistance compared to the MBR. FTIR analysis identified a greater proportion of carbohydrate compounds in the cake layer of the EC-MBR system. SEM images revealed dense microbial clusters, mainly rod- and oval-shaped bacteria, in the EC-MBR system. Furthermore, EDX analysis detected elements such as Ca, K, O, Al, and P in both systems, with the EC-MBR system showing a higher Al content. The EC-MBR system showed low energy consumption (0.431kWhm-3) and total operating costs ($0.90m-3), showcasing its effectiveness and affordability for sustainable wastewater management.

Understanding Bacillus response to salt stress: Growth inhibition, enhanced EPS secretion, and molecular adaptation mechanisms

This study investigates the secretion pattern of extracellular polymeric substances (EPS) by Bacillus sp. under varying salt concentrations and elucidates the molecular mechanisms governing EPS synthesis and secretion. Salt stress inhibited cell proliferation, while optimal salt stimulation promoted EPS secretion, resulting in increased viscosity of the culture medium and the formation of bacterial clusters. Fourier infrared spectrum analysis revealed functional groups such as C-O-C and N-H within the EPS. Soluble-EPS (S-EPS) contained sulfur and phosphorus groups associated with heavy metal ions adsorption. The study also identified a novel polysaccharide formed through bonding EPS (B-EPS). High salt concentrations correlated with elevated levels of tryptophan protein and its derivatives, increased tyrosine polysaccharide derivatives, and decreased aromatic polysaccharides. B-EPS exhibited higher levels of aromatic polysaccharides, with Na+ promoting detachment of B-EPS from the cell surface. Transcriptome sequencing (RNA-seq) analysis under salt stress revealed significant expression of spore kinase (KinD) and response regulatory protein Spo0A in the phosphoric acid relay system. Key transcriptional regulatory factors, including OmpR and exopolysaccharide biosynthesis, were closely associated with EPS synthesis and secretion. This study establishes a theoretical foundation for the industrial production and practical application of EPS by elucidating the molecular mechanisms underlying Bacillus' response to salt stress.

Mechanism of membrane fouling mitigation by microalgae biofilm formation for low C/N mariculture wastewater treatment: EPS characteristics, composition and interfacial interaction energy

Membrane fouling is considered one of the main limitations of membrane bioreactor (MBR) in wastewater treatment applications, including low C/N aquaculture wastewater. Although much research has focused on the integration of MBR with bacterial biofilm, there has been limited exploration into the mitigation mechanism of membrane fouling by microalgae biofilm. In this study, extracellular polymeric substances (EPS) before and after the formation of microalgae biofilm were compared, leading to the conclusion that the microalgae biofilm formation could reduce membrane fouling potential of EPS, thereby mitigating membrane fouling. EPS content and fluorescence components analysis indicated that microalgae biofilm formation could effectively reduce protein to polysaccharide ratios (PN/PS) of three types of EPS (soluble EPS (S-EPS), loosely bound EPS (LB-EPS), and tightly bound EPS (TB-EPS)), lowering the membrane fouling potential associated with protein substances, especially tryptophan-like protein. Moreover, the analysis of molecular weight (MW) suggested that microalgae biofilm could considerably decrease the MW of both S-EPS and LB-EPS, thus mitigating the influence of high MW substances on membrane fouling. Meanwhile, the precise composition of EPS revealed a reduction in hydrophobic alkanes and recalcitrant aromatics, which often lead to membrane fouling. Furthermore, the extended Derjaguin-Landau-Verwey-Overbeek (XDLVO) theory further highlighted that microalgae biofilm weakened the interfacial interaction energy between the three EPS and membrane to mitigate membrane fouling. Therefore, this study provides a comprehensive elucidation of how microalgae biofilm formation mitigates membrane fouling, offering theoretical support for utilizing microalgae biofilm MBR in treating low C/N mariculture wastewater.

Tracking the transformation of extracellular polymeric substances during the ultraviolet/peracetic acid disinfection process: Emphasizing on molecular-level analysis and overlooked mechanisms

In this study, the transformation mechanisms of extracellular polymeric substances (EPS) during ultraviolet/peracetic acid (UV/PAA) disinfection were elucidated based on multiple molecular-level analyses. After UV/PAA disinfection, the contents of soluble EPS (S-EPS), loosely bound EPS (LB-EPS) and tightly bound EPS (TB-EPS) were reduced by 70.47 %, 57.05 % and 47.46 %, respectively. Fluorescence excitation-emission matrix-parallel factor and Fourier transform ion cyclotron resonance mass spectrometry analyses showed that during UV/PAA disinfection, EPS was transformed from the state characterized by high aromaticity, low saturation and low oxidation to the one with reduced aromaticity, increased saturation and higher oxidation. Specifically, sulfur-containing molecules (CHOS, CHONS, etc.) in EPS were converted into highly saturated and oxidized species (such as CHO), with the aromaticity index (AImod) decreasing by up to 53.84 %. Molecular characteristics analyses further indicated that saturation degree, oxidation state of carbon and molecular weight exhibited the most significant changes in S-EPS, LB-EPS and TB-EPS, respectively. Additionally, mechanistic analysis revealed that oxygen addition reaction was the predominant reaction for S-EPS (+O) and TB-EPS (+3O) (accounting for 31.78 % and 36.47 %, respectively), while the dealkylation was the main reaction for LB-EPS (29.73 %). The results were consistent with functional groups sequential responses analyzed by Fourier transform infrared and two-dimensional correlation spectroscopy, and were further verified by density functional theory calculations. Most reactions were thermodynamically feasible, with reaction sites predominantly located at functional groups such as CO, CO, CN and aromatic rings. Moreover, metabolomics analysis suggested that changes in metabolites in raw secondary effluent during UV/PAA disinfection were strongly correlated with EPS transformation. Our study not only provides a strong basis for understanding EPS transformation during UV/PAA disinfection at molecular-level but also offers valuable insights for the application this promising disinfection process.

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.

Interaction between microalgae and phycosphere bacteria in a binary cultivation system-based dairy farm wastewater treatment

The combination of microalgal culture and wastewater treatment is an emerging topic. This study investigated the use of different microalgae to treat different types of dairy farm wastewater. The results showed that the removal of ammonia nitrogen and total phosphorus by mixed microalgae was over 99% and 80%, respectively. The highest production of protein in biomass and extracellular polymeric substances was observed in high-concentration wastewater. In the phycosphere, the abundance of Proteobacteria and Cyanobacteria increased, while that of Bacteroidota decreased. Phycosphere bacteria were strongly correlated with microalgal growth and the composition of extracellular polymeric substances, especially with bound extracellular polymeric substances relative to soluble extracellular polymeric substances. Genes associated with photosynthesis and respiration in phycosphere bacteria were upregulated, contributing to the material exchange capacity in the microalgal-bacterial systems. The interaction between microalgae and phycosphere bacteria thus represents the core of the binary cultivation system-based wastewater treatment and requires further investigation.

Mitigation of fouling in membrane bioreactor using the integration of electrical field and nano chitosan adsorbent

The application of membrane bioreactors (MBRs) has emerged as an impressive solution to water scarcity. One of the main obstacles for MBR application is membrane fouling. This work studied the effect of the novel simultaneous application of electric field and positively-charged nano chitosan on membrane fouling. Four MBRs of 1 (control bioreactor), 2 (control bioreactor + electric filed), 3 (control bioreactor + nano chitosan) and 4 (control bioreactor + electric filed + nano chitosan) were evaluated. The results indicated that the concurrent use of voltage and nano chitosan better reduced the membrane fouling, decreased flux decline, and enhanced recovery ratios and membrane bioreactor performance index by nearly 43 % and 84 %, respectively. Meanwhile, the contents of soluble microbial products (SMP) and extracellular polymeric substances (EPS) were reduced from MBR1 to MBR4, respectively. Reduced zeta potential and increased particle sizes led to the change in the cake layer and fouling mitigation from MBR1 to MBR4, as observed through scanning electron microscope (SEM) images. Excitation-emission matrix (EEM) analysis showed that humic acid was adsorbed by both electro-coagulants and nano adsorbents, resulting in a substantial (approximately 97 %) reduction in membrane pore fouling for MBR4. The combination of electrocoagulation, electro-oxidation, and electrophoresis processes, along with adsorption by positively charged nano chitosan, effectively mitigated membrane fouling. The cathode induced a negative charge on sludge particles, subsequently facilitating their adsorption by the positively charged nano chitosan adsorbents. The results demonstrated that combining electric field and nano chitosan effectively suppresses membrane fouling, with relevance for forthcoming technological advancements.

Investigating the Dewatering Efficiency of Sewage Sludge with Optimized Ratios of Electrolytic Manganese Residue Components.

As an industrial waste residue, Electrolytic Manganese Residue (EMR) can greatly promote sludge dewatering and further particle-size optimization can significantly strengthen sludge dewaterability. In this study, the effects of ammonium sulfate, calcium sulphate dihydrate, and manganese carbonate in EMR on sludge dewatering performance were investigated using the response surface optimization method. It was found that the optimized ratio of three components in EMR was 1.0:1.6:2.2 based on capillary suction time (CST), specific resistance of filtration (SRF), and zeta potential of dewatered sludge. The composition ratio of particle-size optimized EMR was modified based on the above optimization, resulting in a significant increase in sludge dewatering performance (CST and SRF reduced by 8.7% and 11.2%, respectively). Compared with those in original sludge, the content of bound extracellular polymeric substances in the conditioned sludge with optimized ratio was drastically reduced while that of soluble extracellular polymeric substances was slightly increased, which was in accordance with the decline of fluorescence intensity. These findings indicated the disintegration of extracellular polymeric substances, the enhancement of hydrophobicity, and dewatering properties of the sludge. In summary, optimized EMR can effectively intensify the dewaterability of sludge, providing a competitive solution for dewatering and further disposal of sludge.

Mechanisms of conjugative transfer of antibiotic resistance genes induced by extracellular polymeric substances: Insights into molecular diversities and electron transfer properties

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.

Controlling of irreversible fouling and mechanism in a hybrid ceramic membrane bioreactor (CMBR)-reverse osmosis (RO) process for textile wastewater reclamation

In this study, the fouling propensity of a hybrid ceramic membrane bioreactor (CMBR)-reverse osmosis (RO) process was investigated over a prolonged period using real textile wastewater effluent. Additionally, the effectiveness of in-situ ozonation for fouling mitigation was investigated. The results revealed that the CMBR-RO system achieved approximately 99.8 % removal of chemical oxygen demand and 97.4 % removal of total nitrogen. Despite this, soluble extracellular polymeric substances, particularly biopolymers and humics-like substances, remained in the system, resulting in irreversible fouling on the ultrafiltration (UF) membrane. The implementation of in-situ ozonation notably reduced ceramic membrane fouling by enhancing the filterability of larger macromolecules and eliminating inorganic/organic foulants from the membrane surface. Multiple modified fouling indexes were also utilized to evaluate the fouling potential of the RO feed water through size fractionation. It is worth noting that ozonation could pose a potential risk for subsequent RO membrane fouling due to the formation of low molecular weight neutrals, acids, and calcium- or silica- related inorganic-organic complexes. Overall, the integrated CMBR-RO system with in-situ ozonation exhibited excellent performance in producing clean water, achieving nearly 100 % removal of 20 Aromatic Amines. These findings underscore the viability of the CMBR-RO system as a practical and sustainable choice for the reclamation of textile wastewater.

The effect of temperature on fouling in anaerobic membrane bioreactor: SMP- and EPS-membrane interactions

Biofouling is the main challenge in the operation of anaerobic membrane bioreactors (AnMBRs). Biofouling strongly depends on temperature; therefore, we hypothesize that the interactions and viscoelastic properties of soluble microbial products (SMP) and extracellular polymeric substances (EPS) vary with temperature, consequently influencing membrane permeability. This study compares the performance of an AnMBR operated at a similar permeate flux at two temperatures. The transmembrane pressure (TMP) rose rapidly after 5 ± 2 days at 25 °C but only after 18 ± 2 days at 35 °C, although the reactor's biological performance was similar at both temperatures, in terms of the efficiency of dissolved organic carbon removal and biogas composition, which were obtained by changing the hydraulic retention time. Using confocal laser scanning microscopy (CLSM), a higher biofilm amount was detected at 25 °C than at 35 °C, while quartz crystal microbalance with dissipation (QCM-D) showed a more adhesive, but less viscous and elastic EPS layer. In situ optical coherence tomography (OCT) of an ultra-filtration membrane, fed with the mixed liquor suspended solids (MLSS) at the two temperatures, revealed that while a higher rate of TMP increase was obtained at 25 °C, the attachment of biomass from MLSS was markedly less. Increased EPS adhesion to the membrane can accelerate TMP increase during the operation of both the AnMBR and the OCT filtration cell. EPS's reduced viscoelasticity at 25 °C suggests reduced floc integrity and possible increased EPS penetration into the membrane pores. Analysis of the structures of the microbial communities constituting the AnMBR flocs and membrane biofilms reveals temperature's effects on microbial richness, diversity, and abundance, which likely influence the observed EPS properties and consequent AnMBR fouling.

Preliminary tests on carbon and nitrogen emissions and nutrients availability upon application of algal-bacterial granules to arid and low fertility soil

To remediate arid and low fertility soil and explore the feasibility of controlling its carbon (C) and nitrogen (N) emissions, algal-bacterial aerobic granular sludge (AGS) was applied as soil additives and compared with bacterial AGS and cyanobacteria during 14 days’ incubation. Results show that algal-bacterial AGS as additives possesses a higher potential in terms of fertility recovery and soil physicochemical properties conservation. The additives may change the soil characteristics by decreasing soil pH (within 0.6) and water content (0.8–2.5 %) with increased electrical conductivity during the 14 days’ incubation. As for soil fertility, due to the high available phosphorus (P) content in algal-bacterial AGS, the soil available P content was significantly increased from 50 % to 170 % after incubation under different application rates or gaseous environments. N was also accumulated in the soil matrix with increased inorganic forms and less or acceptable N emission. The contents of soluble organic substances and extracellular polymeric substances (EPS) were significantly increased after incubation. Under ambient air and moderately elevated CO2 like 10 % conditions, carbon mineralization in the soil samples was lower than 15 % at low additives dosages; while under an elevated CO2 content (25 %) condition, a higher additives dosage may cause a rapid CO2 accumulation within 2 days with a substantial O2 consumption, leading to an anaerobic/anoxic environment corresponding to a higher C mineralization and/or N loss.

Predatory Bdellovibrio-and-like organism mixtures on efficient MBR in-situ membrane fouling diminishment and mechanisms

ABSTRACT Membrane fouling is a major hindrance that restricts the application of membrane bioreactors (MBRs). Bdellovibrio-and-like organisms (BALOs), as obligatory parasitic bacteria, prey upon various bacteria. In this study, the BALO mixtures were screened and found more effective in membrane fouling mitigation compared to the single BALO species and extended the membrane filtration period by as long as 33.3%. The higher BALO diversity reduced the potential foulants generation in the activated sludge by decreasing the sludge viscosity as high as 13.8 ± 0.6% than the pure culture of BALO. Meanwhile, the mixed BALOs demonstrated superior biofilm predation capabilities, with the content of soluble microbial products and extracellular polymeric substances on the biofilm decreasing by 26.1 ± 0.5% and 38.3 ± 0.2% as the most compared to the single BALO species involved system. Additionally, the BALO mixtures expanded the single strains’ host lysis spectrum of both the activated sludge and biofilm. The abundance of membrane-fouling-related bacteria such as Flavobacterium, Rhodobacter, and Labilithrix and pioneer bacteria such as Sphingorhabdus and Pseudomonas was significantly reduced. In summary, this study disclosed the significantly better membrane fouling mitigation effects of the BALOs with higher diversity, suggesting that the expansion of the host range is crucial for the further application of BALOs to enhance the anti-fouling performance of the MBR system.

Effect of ciprofloxacin on the one-stage partial nitrification and anammox biofilm system: A multivariate analysis focusing on size-fractionated organic components

The impact of ciprofloxacin (CIP) in the partial nitrification and anammox biofilm system was investigated by multivariate analysis, focusing on size-fractionated organic components. The CIP dose of 10 μg/L did not inhibit the total nitrogen (TN) removal efficiency, even though the abundance of antibiotic resistant genes (ARGs) (i.e., qnrD, qnrB, qnrA, qnrS, and arcA) was elevated. However, a gradual higher CIP dosing up to 100 μg/L inhibited the TN removal efficiency, while the abundance of ARGs was still increased. Moreover, both the TN removal efficiency and the abundant ARGs were dwindled at 470 μg/L of CIP. As the CIP dose increased from 0 to 100 μg/L, the abundance of high molecular weight (MW) fractions (14,000 to 87,000 Da; 1000 to 14,000 Da) and humic/fulvic acid-like components in the soluble extracellular polymeric substances (HSS) decreased, with more increases of low MW (84–1000 Da; less than 84 Da) fractions and soluble microbial by-products in soluble extracellular polymeric substances (SMPS). Continuously increasing the CIP dose till 470 μg/L, an inverse trend of the changes of these organic components was noted, along with clear reductions of the microbial diversity and richness, and the abundance of key functional genes responsible for nitrogen removal. The predominance of functional gene amoA (related with ammonia oxidizing bacteria) was more significantly with more distribution of SMPS with relatively low MW and less distribution of HSS with relatively high MW, as well as polymer decomposing microorganisms such as Bryobacteraceae and the unclassified Saprospirales.

Enhancing short-chain fatty acids production via acidogenic fermentation of municipal sewage sludge: Effect of sludge characteristics and peroxydisulfate pre-oxidation.

This study first employed a combined pretreatment of low-dose peroxy-disulfate (PDS) and initial pH 10 to promote short-chain fatty acids (SCFAs) production via acidogenic fermentation using different types of sewage sludge as substrates. The experimental results showed that the yield of maximal SCFAs and acetate proportion after the combined pretreatment were 1513.82±28.25mg chemical oxygen demand (COD)/L and 53.64%, and promoted by 1.28 and 1.56 times higher, respectively, compared to the sole initial pH 10 pretreatment. Furthermore, in terms of the disintegration degree of sewage sludge, it increased by more than 18% with the combined pretreatment compared to the pretreatment of sole initial pH 10. Waste-activated sludge (WAS) from A2/O and Bardenpho processes were more biodegradable, explained by the 1.47- and 1.35-times higher disintegration rate than those from oxidation ditch and they favored acetate dominant fermentation. Correlation analysis revealed a strong correlation (p ≤ 0.01) between SCFAs production and soluble COD, total proteins, proteins in soluble-extracellular polymeric substances (SEPS), total polysaccharides, and polysaccharides in SEPS. Mechanism explorations showed that preoxidation with PDS enhanced the solubilization and biodegradability of complex substrates, and altered the microbial community structure during the fermentation process. Firmicutes and Tetrasphaera were proven to play a key role in improving SCFA production, especially in promoting acetate production by converting additional SCFAs into acetate. Additionally, the addition of PDS greatly promoted sulfur and iron-related metabolic activities. Finally, the combined pretreatment was estimated to be a cost-effective solution for reutilizing and treating Fe-sludge.

A step towards more eco-friendly and efficient microalgal harvesting: Inducing flocculation in the non-naturally flocculating strain Chlorella vulgaris (13-1) without chemical additives

Flocculation is often regarded as a cost-effective and reliable method for microalgal harvesting. However, the traditional method often requires the addition of chemical agents to induce flocculation. This carries certain disadvantages including the chemical contamination of the biomass and the subsequent need to remove the flocculants from the medium. To address these issues, this study aimed to induce flocculation in a naturally non-flocculating strain (Chlorella vulgaris 13-1) without resorting to chemical additives, with the ultimate goal of increasing harvesting efficiency. Scanning electron microscopy showed that Scotelliopsis reticulata UFA-2, a naturally flocculating strain, produces extracellular polymeric substances (EPS) whereas 13-1 does not. As a result, two methods were used to induce flocculation in 13-1: co-cultivation of UFA-2 and 13-1, and insertion of EPS produced by UFA-2 into the growth medium of 13-1. The co-cultivation of 13-1 with UFA-2 significantly increased the flocculation efficiency compared to that of 13-1 alone (30 % higher flocculation efficiency after one hour of settling and 52 % higher after three hours of settling). Alternatively, the insertion of dry tightly-bound (TB) UFA-2 EPS into 13-1 cultures also improved flocculation efficiency (by 19 % compared to the control), while addition of soluble or loosely-bound (LB) EPS was less efficient (less than 1 % and 10 %, respectively). FTIR results showed that the composition of TB-EPS was different to that of LB and soluble EPS. TB-EPS contained higher proportions of proteins and different types of carbohydrates, potentially contributing to its increased efficacy in inducing flocculation in microalgae.

Interface Engineering of PdPt Ultrafine Ethanol Electro-Oxidation Nanocatalysts by Bacterial Soluble Extracellular Polymeric Substances (s-EPS) to Break through Sabatier Principle.

Unsatisfactory performance of ethanol oxidation reaction (EOR) catalysts hinders the application of direct ethanol fuel cells (DEFCs), while traditional alloy catalysts (like PdPt) is cursed by Sabatier principle due to countable active site types. However, bacterial soluble extracellular polymeric substances (s-EPS) owning abundent functional groups may help breacking through it by contrusting different active sites on PdPt and inducing them to play synergy effect, which is called interface engineering. Using s-EPS to engineer catalysts is more green and consumes lower energy compared to chemical reagents. Herein, PdPt alloy nanoparticles (≈2.1nm) are successfully in situ synthesized by/on s-EPS of Bacillus megaterium, an ex-holotype. Tryptophan residuals are proved as the main reductant. In EOR, PdPt@s-EPS shows higher activity (3.89mAcm-2) than Pd@s-EPS, Pt@s-EPS, Pt/C and most reported akin catalysts. Its stability and durability are excellent, too. DFT modelling further demonstrates that, interface engineering by s-EPS breaks through Sabatier principle, by the synergy of diverse sites owning different degrees of d-p orbital hybridization. This work not only makes DEFCs closer to practice, but provides a facile and green strategy to design more catalysts.

Biodegradation of phenanthrene-Cr (VI) co-contamination by Pseudomonas aeruginosa AO-4 and characterization of enhanced degradation of phenanthrene

Microorganisms capable of simultaneously remediating heavy metals (HMs) and polycyclic aromatic hydrocarbons (PAHs) pollution hold significant importance in bioremediation efforts. In this study, we investigated the ability of Pseudomonas aeruginosa AO-4 to simultaneously degrade phenanthrene (PHE) and reduce Cr (VI). Specifically, it has the ability to reduce 100 % of Cr (VI) (30 mg/L) while degrading 43.8 % of PHE (50 mg/L). In batch experiments, it was observed that the presence of Cr (VI) can enhance the degradation of PHE by strain AO-4. The solubility of PHE in soluble extracellular polymeric substances (S-EPS) was found to be related to the initial concentration of Cr (VI), which could explain why Cr (VI) promotes the degradation of PHE. Additionally, XPS analysis confirmed that Cr (VI) was reduced to Cr (III) with S-EPS produced by Pseudomonas aeruginosa AO-4. GC–MS analysis was conducted to analyze the degradation metabolites of phenanthrene, di(2-ethylhexyl) phthalate and 2TMS derivatives of salicylic acid were detected, indicating that Pseudomonas aeruginosa AO-4 is capable of degrading phenanthrene through two distinct pathways. These findings demonstrate the potential of Pseudomonas aeruginosa AO-4 in the treatment of co-contamination scenarios involving PAHs and HMs.