Total Extracellular Polymeric Substances Research Articles

Enhancing sulfide and methane control in sewers: Unveiling the impact and mechanisms of free nitrous acid pre-exposure coupled with calcium peroxide dosing

Although biocides have proven highly efficient in controlling hazardous emissions and mitigating organic carbon loss in sewers, the resistance of biofilm and extracellular polymeric substances (EPS) to biocides has emerged as a significant impediment to addressing these challenges. In this study, a promising strategy of calcium peroxide (CaO2) dosing assisted by free nitrous acid (FNA) pre-exposure overcame these limitations. Prior to a 6-hour exposure to 0.1 % (w/v) CaO2, a 2-hour pre-exposure to a low dose of 0.26 mg-N/L FNA demonstrated a significant enhancement in sulfide and methane control efficiency by 267.14 % and 390.47 %, respectively, compared to direct exposure to 0.2 % CaO2 for 12 h. Moreover, there was a notable increase of 40.05 % in soluble COD level in the effluent. FT-IR and XPS analyses discovered the disruption, migration, and dissolution of biofilm, accompanied by a substantial increase in membrane permeability (up to 2.89 folds). Additional analysis revealed that the synergistic effects of various reactive nitrogen/oxygen species destabilized the structure of EPS, e.g., declining total EPS yield, disintegrating humic acid-like and tryptophan- and protein-like substances, reducing hydrogen bond in β-sheet of proteins. Further down-regulation of node genes associated with sulfidogenesis and methanogenesis verified the toxic mechanism at the genetic level. Microbial community analysis revealed a significant reduction in the abundance of sulfate-reducing bacteria and methanogenic archaea, leading to an extended recovery period for sulfide and methane production in sewer biofilm. This study provides an effective strategy and in-depth mechanisms for biofilm control, applicable beyond sewer biofilm.

Mechanisms of partial denitrification in an integrated fixed-film activated sludge (IFAS) system: From progressive startup to stabilization

The rapid startup and long-term stability of partial denitrification (PD) processes are essential prerequisites for successful PD/Anammox systems. Bioaggregates play a significant role in impacting mass transfer efficiency and shaping the microbial community structure within the PD process. Thus, the startup and stable operation of the PD process were investigated in an integrated fixed-film activated sludge (IFAS) by inoculating activated sludge and biofilm from anoxic tanks of a wastewater treatment plant. During the initial stage of startup, a sudden increase in NO3−-N concentration induced a rapid enzyme stress response, leading to a significant increase in total NaR activity (1.38 ± 0.0 μmol·(L·min)−1 to 25.94 ± 0.23 μmol·(L·min)−1), concomitant with the inhibition of total NiR activity (13.93 ± 0.48 μmol·(L·min)−1 to 4.25 ± 0.26 μmol·(L·min)−1), thereby inducing the rapid accumulation of NO2−-N (13.39 mg·L−1) within 6 days. During the later stage of startup, enzyme stress response was subsided but the partial denitrifiers were gradually enriched, consequently leading to continuous accumulation of NO2−-N. The results of ex-situ activity showed that flocs exhibit superior PD performance in contrast to biofilm throughout the entire operational period. Meanwhile, the total extracellular polymeric substances (EPS) content and community structure in both flocs and biofilm indicated that the migration of EPS-secreted partial denitrifiers (Thauera) from flocs to biofilm was obliged by substrate deficiency during the stable operating stage (days 35–100). Therefore, the startup of PD may be sequentially mediated by two distinct mechanisms, enzyme stress response and functional bacterial enrichment, with microbial community migration occurring during the stable phase.

Low-biofouling membrane bioreactor: Effects of cis-2-Decenoic acid addition on EPS and biofouling mitigation

Biofouling is inevitable in the membrane process, particularly in membrane bioreactors (MBR) combined with activated sludge processes. Regulating microbial signaling systems with diffusible signal factors such as cis-2-Decenoic acid (CDA) can control biofilm formation without microbial death or growth inhibition. This study assessed the effectiveness of CDA in controlling biofouling in membrane bioreactors (MBRs), essential for wastewater treatment. By modulating microbial signaling, CDA mitigated biofilm formation without hindering microbial growth. Analysis using Confocal Laser Scanning Microscopy (CLSM) revealed structural alterations in the biofilm, reducing biomass and thickness upon CDA application. Moreover, examination of extracellular polymeric substances (EPS) highlighted a decrease in total EPS, particularly effective polysaccharides. In addition, the possibility of shifting from high molecular weight EPS to low molecular weight EPS was revealed through the change in dispersion activity. The 56% extension of MBR operational lifespan resulting from the reduction in EPS is anticipated to offer potential cost savings and improved performance. Despite these results, further investigation is crucial to validate any potential environmental risks associated with CDA and to comprehend its long-term effects at various conditions.

Differentiation and quantification of extracellular polymeric substances from microalgae and bacteria in the mixed culture

Extracellular polymeric substances (EPS) play significant roles in the formation, function, and interactions of microalgal-bacteria consortia. Understanding the key roles of EPS depends on reliable extraction and quantification methods, but differentiating of EPS from microalgae versus bacteria is challenging. In this work, cation exchange resin (CER) and thermal treatments were applied for total EPS extraction from microalgal-bacteria mixed culture (MBMC), flow cytometry combined with SYTOX Green staining was applied to evaluate cell disruption during EPS extraction, and auto-fluorescence-based cell sorting (AFCS) was used to separate microalgae and bacteria in the MBMC. Thermal extraction achieved much higher EPS yield than CER, but higher temperature and longer time reduced cell activity and disrupted the cells. The highest EPS yield with minimal loss of cell activity and cell disruption was achieved using thermal extraction at 55℃ for 30 min, and this protocol gave good results for MBMC with different microalgae:bacteria (M:B) mass ratios. AFCS combined with thermal treatment achieved the most-efficient biomass differentiation and low EPS loss (<4.5 %) for the entire range of M:B ratios. EPS concentrations in bacteria were larger than in microalgae: 42.8 ± 0.4 mg COD/g TSS versus 9.19 ± 0.38 mg COD/g TSS. These findings document sensitive and accurate methods to extract and quantify EPS from microalgal-bacteria aggregates.

Glycine betaine modulates extracellular polymeric substances to enhance microbial salinity tolerance

High salinity inhibits microbial activity in the bioremediation of saline wastewater. To alleviate osmotic stress, glycine betaine (GB), an osmoprotectant, is added to enhance the secretion of extracellular polymeric substances (EPS). These EPS are pivotal in withstanding environmental stressors, yet the intricate interplay between GB supplementation and microbial responses through EPS modifications—encompassing composition, molecular architecture, and electrochemical features—remains elusive in hypersaline conditions. Here we show microbial strategies for salinity endurance by investigating the impact of GB on the dynamic alterations of EPS properties. Our findings reveal that GB supplementation at 3.5% salinity elevates the total EPS (T-EPS) content from 12.50 ± 0.05 to 24.58 ± 0.96 mg per g dry cell weight. The observed shift in zeta potential from −28.95 to −6.25 mV at 0% and 3.5% salinity, respectively, with GB treatment, indicates a reduction in electrostatic repulsion and compaction. Notably, the EPS protein secondary structure transition from β-sheet to α-helix, with GB addition, signifies a more compact protein configuration, less susceptible to salinity fluctuations. Electrochemical analyses, including cyclic voltammetry (CV) and differential pulse voltammetry (DPV), reveal GB's role in promoting the release of exogenous electron shuttles, such as flavins and c-type cytochromes (c-Cyts). The enhancement in DPV peak areas (QDPV) with GB addition implies an increase in available extracellular electron transfer sites. This investigation advances our comprehension of microbial adaptation mechanisms to salinity through EPS modifications facilitated by GB in saline habitats.

Operating performance of sequential batch system with calcium-modified basalt fiber bio-nest exposed to two typical perfluoroalkyl compounds (PFOA and PFBA)

The effects of per- and polyfluoroalkyl substances (PFASs) on the application of sequential batch system with basalt fiber as a biological carrier material have been rarely reported. This study explored the decontamination performance, extracellular polymeric substances (EPS) and microbial community structure of sequencing batch system with calcium-modified basalt fiber (Ca-MBF) bio-nest exposed to perfluorooctanoic acid (PFOA) and perfluorobutyric acid (PFBA). The results showed that 0.5 mg/L PFOA and PFBA both inhibited nitrification by 24.67 % and 19.47 %, while promoted denitrification by 16.54 % and 11.15 %, respectively. Overall, compared to PFBA, PFOA inhibited the removal of total nitrogen (TN) and total phosphorus (TP) by 11.27 % and 9.49 %, respectively. Further analysis of EPS showed that under PFOA exposure, the total EPS and polysaccharide (PS) content decreased by 34.57 % and 40.78 %, respectively. In addition, PFOA and PFBA exhibited a phenomenon of initially promoting by 89.21 % and 55.29 %, and subsequently inhibiting on protein (PN) content by 50.22 % and 51.37 %, respectively. Analyses of Illumina Miseq sequencing showed that PFOA and PFBA exposure for 50 days both inhibited the richness and diversity of microbial communities, and the inhibitory effect of PFOA was greater than that of PFBA. In addition, PFOA and PFBA respectively domesticated the resistant bacteria Meganema and Acinetobacter. These results may provide valuable information for ecological restoration and biodegradation of PFOA and PFBA.

Larger hydroxyapatite aggregation from Ca2+ adhesion in ANAMMOX granular sludge caused by high dissolved oxygen

Anaerobic ammonia oxidation (ANAMMOX), a sustainable biological process, is promising to remove NH4+-N from municipal sewage. In this study, results showed that the anammox granular sludge morphology changes with the alternation of dissolved oxygen (DO), mainly attributing to the adhesion of calcium ions (Ca2+) to the surface of sludge particles. Diverse characterization methods revealed that gray adhesions in the form of hydroxyapatite covered the original holes on the anammox granular sludge surface, including scanning Electron Microscopy (SEM), digital camera images, Energy Dispersive Spectrometer (EDS), and X-ray diffraction (XRD). Ex-situ degradation of NH4+-N and NO2−-N yielded diverse outcomes. The protein to polysaccharide ratio (PN/PS) in the total extracellular polymeric substances (EPS) across 4 size groups demonstrated a decrease under O2 exposure. Microbial community analysis indicated norank_f_A4b and Nitrolancea being the most abundant genus under O2 exposure at day 1 and day 100, respectively. These findings offer an effective strategy to prevent size-larger granular sludge from deteriorating through changing DO and Ca2+ in municipal wastewater in ANAMMOX.

Impacts of Polylactic Acid Microplastics on Performance and Microbial Dynamics in Activated Sludge System

A large number of microplastics (MPs) have been found in various stages of wastewater treatment plants, which may affect the functional microbial activity in activated sludge and lead to unstable pollutant removal performance. In this study, the effects of different concentrations of polylactic acid microplastics (PLA MPs) on system performance, nitrification and phosphorus (P) removal activities, and extracellular polymeric substances (EPS) were evaluated. The results showed that under the same influent conditions, low concentrations (50 particles/(g TS)) of PLA MPs had no significant effect on effluent quality. The average removal efficiencies of chemical oxygen demand, phosphate, and ammonia were all above 80%, and the average removal efficiencies of total nitrogen remained above 70%. High concentrations (200 particles/(g TS)) of PLA MPs inhibited the activities of polyphosphate-accumulating organisms (PAOs) and nitrifying bacteria. The specific anaerobic P release rate decreased from 37.7 to 23.1 mg P/(g VSS·h), and the specific aerobic P uptake rate also significantly decreased. The specific ammonia oxidation rate decreased from 0.67 to 0.34 mg N/(g VSS·h), while the change in the specific nitrite oxidation rate was not significant. The dosing of PLA MPs decreased the total EPS and humic acid content. As the concentration of PLA MPs increased, microbial community diversity increased. The relative abundance of potential PAOs (i.e., Acinetobacter) increased from 0.08 to 12.57%, while the relative abundance of glycogen-accumulating organisms (i.e., Competibacter and Defluviicoccus) showed no significant changes, which would lead to improved P removal performance. The relative abundance of denitrifying bacteria (i.e., Pseudomonas) decreased from 95.43 to 58.98%, potentially contributing to the decline in denitrification performance.

Enhanced disintegration mechanism of surplus activated sludge to improve dewatering by thermally activated persulfate oxidation under mild temperature.

The dewatering treatment is an essential process for the treatment and disposal of surplus activated sludge (SAS), and improving sludge dewatering performance is still a challenge and has become a research hotspot in recent years. The oxidation and disintegration of bacterial cells and extracellular polymeric substances (EPS) by active radicals produced by advanced oxidation processes (AOPs) were extremely promising to achieve deep sludge dewatering. This paper systematically studied the efficiency and mechanism of thermally activated persulfate (TAP) oxidation technology to the improvement of SAS dewatering performance. The results showed that the relative filterability (CST0/CST) was increased 2.52 times with 2.0 mmol/g VSS potassium peroxydisulfate (PDS) at 80 °C in 90 min. Under this condition, the Zeta potential of SAS significantly decreased from - 14.8 to - 1.44 mV, while the average particle size (dp50) decreased from 52.981 to 48.259 μm. Thermal treatment disrupted the sludge structure to release large amounts of EPS including polysaccharides and protein. Meanwhile, the results of three-dimensional excitation-emission matrix (3D-EEM) fluorescence spectra showed that the TAP treatment could expedite the disintegration of sludge, facilitating the decrease of total EPS content and conversion of tightly bound EPS (TB-EPS) to loosely bound EPS (LB-EPS) and soluble EPS (S-EPS). The mechanism of TAP process to improve SAS dewatering performance was revealed, which could contribute to breaking the bottleneck of sludge depth dewatering and provide a theoretical and technical basis for its practical application.

Sequential adaptation strategies of SPDA systems to low temperature: EPS mediation and community structure evolution

Temperature stands as a vital factor in the practical application of simultaneous partial-denitrification/anammox (SPDA) systems in mainstream wastewater treatment, thus it is necessary to understand the adaptation strategy of SPDA systems to low temperature. To address this, the potential adaptation of different coupling forms to low temperatures was investigated in a moving bed biofilm reactor (MBBR) and an integrated fixed-film activated sludge (IFAS) reactor for 182 days. The results showed that IFAS reactor achieved higher total nitrogen removal rates (TNRRs) at 15 °C (89.67 ± 0.01%) than MBBR system (77.28 ± 0.02%). Nitrogen conversion in the typical cycles showed that PD process was more sensitive to low temperatures than anammox process in SPDA systems. Meanwhile, the significant increase of total extracellular polymeric substances (EPS) content in each biophase and the change of community structure indicated that EPS mediation and community structure evolution in different biophases are two adaptive strategies of SPDA system to temperature decreasing. Biofilm phase tends to produce more EPS to resist the temperature decrease than floc phase. The highly negative relationship between EPS content at 15 °C and the initial temperature of change in community structure (p = 0.036＜0.05, r = -0.964) suggested that microorganisms always tend to resist the adverse environment through EPS mediation first, and then adapt to the low-temperature environment through community structure evolution with the temperature further decreased. This study guided the selection of SPDA coupling system in practical engineering applications, and laid a foundation for efficient biological nitrogen removal at low temperatures.

The effect of crude oil on hydrolyzed polyacrylamide-containing wastewater treatment using microbial fuel cell biosystem

One important way to recover valuable energy from the waste streams is to convert organic wastewater into bioelectricity. Microbial fuel cell (MFC) served as the green and sustainable biological treatment can simultaneously treat wastewater and produce moderate amounts of electricity. In the actual hydrolyzed polyacrylamide (HPAM)-containing wastewater of oilfield, crude oil is served as the important active components that affects HPAM bioconversion in biological treatment system. The effect of different crude oil levels on the bioelectric conversion of HPAM in MFC biosystem has not been explored. This study is the first to explore the bioelectric transformation of HPAM-containing wastewater in MFC biosystem under the stress of different initial crude oil levels. The output voltage, current density, coulombic efficiency and power density showed an upward tendency as crude oil level increased (0–300 mg·L−1), and their maximums reached 22.7 V, 267 mA·m−2, 59.6% and 2.42 W·m−2, respectively. The elevation of crude oil level improved HPAM biodegradation, total nitrogen (TN) removal, metabolite production and extracellular polymeric substance (EPS) intensity, and enhanced carbon/nitrogen bioconversion and electron transfer. Geobacter, Pseudomonas, Flavobacterium, Desulfosalsimonas and Acinetobacter exhibited both substrate degradation and bioelectrochemical activity, dominating the biosystem functions associated with bioelectricity generation, substrate bioconversion, metabolite production and EPS intensity. The elevation of crude oil level stimulated the abundance and function of functional bacteria. This study is of dual practical significance for bioenergy recovery and environmentally friendly bioconversion of HPAM-containing oilfield wastewater, promoting the ecological management of oilfields.

Metagenomics and proteomics profiling of extracellular polymeric substances from municipal waste sludge and their application for soil and water bioremediation

This study assessed the components of anaerobically digested sludge, activated sludge, and microbial and extracellular polymeric substance (EPS) enzymes to identify the mechanisms underlying nitrogen removal and soil regeneration. 16S rRNA gene amplicon-based sequencing was used to determine the microbial community composition and the related National Center for Biotechnology Information (NCBI) protein database was used to construct a conventional library from the observed community. EPS components were identified using gel-free proteomic (Liquid Chromatography with tandem mass spectrometry-LC/MS/MS) methods. Alginate-like EPS from aerobically activated sludge have strong potential for soil aggregation and water-holding capacity, whereas total EPS from anaerobic sludge have significant potential for ammonia removal under salt stress. Fourier transform infrared spectroscopy (FTIR) revealed that both EPS may contain proteins, carbohydrates, humic compounds, uronic acid, and DNA and determined the presence of O–H, N–H, C–N, CO, and C–H functional groups. These results demonstrate that the overall enzyme activity may be inactivated at 30 g L-1 of salinity. An annotation found in Kyoto Encyclopedia of Genes and Genomes (KEGG)- KEGG Automatic Annotation Server (KAAS) revealed that the top two metabolic activities in the EPS generated from the anaerobic sludge were methane and nitrogen metabolism. Therefore, we focused on the nitrogen metabolism reference map 00910. EPS from the anaerobically digested sludge exhibited nitrate reductase, nitrite reductase, and dehydrogenase activities. Assimilatory nitrate reduction, denitrification, nitrification, and anammox removed ammonia biochemically. The influence of microbial extracellular metabolites on water-holding capacity and soil aggregation was also investigated. The KAAS-KEGG annotation server was used to identify the main enzymes in the activated sludge-derived alginate-like extracellular EPS (ALE-EPS) samples. These include hydrolases, oxidoreductases, lyases, ligases, and transporters, which contribute to soil fertility and stability. This study improves our understanding of the overall microbial community structure and the associated biochemical processes, which are related to distinct functional genes or enzymes involved in nitrogen removal and soil aggregation. In contrast to conventional methods, microbial association with proteomics can be used to investigate ecological relationships, establishments, key player species, and microbial responses to environmental changes. Linking the metagenome to off-gel proteomics and bioinformatics solves the problem of analyzing metabolic pathways in complex environmental samples in a cost-effective manner.

Understanding the cometabolic degradation of sulfadiazine by an enriched ammonia oxidizing bacteria culture from both extracellular and intracellular perspectives

Antibiotics are widely used drugs in the world and pose serious threats to ecosystems and human health. Although it has been reported that ammonia oxidizing bacteria (AOB) can cometabolize antibiotics, little has been reported on how AOB would respond to the exposure of antibiotics on extracellular and enzymatic levels, as well as the impact of antibiotics on the bioactivity of AOB. Therefore, in this study, a typical antibiotic, sulfadiazine (SDZ), was selected, and a series short-term batch tests using enriched AOB sludge were conducted to investigate the intracellular and extracellular responses of AOB along the cometabolic degradation process of SDZ. The results showed the cometabolic degradation of AOB made the main contribution to SDZ removal. When the enriched AOB sludge was exposed to SDZ, ammonium oxidation rate, ammonia monooxygenase activity, adenosine triphosphate concentration and dehydrogenases activity were negatively affected. The amoA gene abundance increased 1.5 folds within 24 h, which may enhance the uptake and utilization of substrates and maintain stable metabolic activity. In the tests with and without ammonium, the concentration of total EPS increased from 264.9 to 231.1 mg/gVSS to 607.7 and 538.2 mg/gVSS, respectively, under the exposure to SDZ, which was mainly contributed by the increase of proteins in tightly bound extracellular polymeric substances (EPS) and polysacharides in tightly bound EPS and soluble microbial products. The proportion of tryptophan-like protein and humic acid-like organics in EPS also increased. Moreover, SDZ stress stimulated the secretion of three quorum sensing signal molecules, C4-HSL (from 140.3 to 164.9 ng/L), 3OC6-HSL (from 17.8 to 42.4 ng/L) and C8-HSL (from 35.8 to 95.9 ng/L) in the enriched AOB sludge. Among them, C8-HSL may be a key signal molecule that promoted the secretion of EPS. The findings of this study could shed more light on the cometabolic degradation of antibiotics by AOB.

How does Microcystis aeruginosa respond to elevated temperature?

Cyanobacteria and their toxins widely exist in freshwater ecosystems. Microcystis aeruginosa is among dominant bloom-forming cyanobacteria. Water temperature is a key factor influencing the life cycle of M. aeruginosa. We simulated elevated temperature (4–35 °C) experiment and cultured M. aeruginosa during the overwintering, recruitment and rapid growth phases. The results showed that M. aeruginosa recovered growth after overwintering at 4–8 °C and recruited at 16 °C. The total extracellular polymeric substance (TEPS) concentration increased rapidly at 15 °C. The actual quantum yield of photosystem II (Fv′/Fm′) peaked at 20 °C during the rapid growth phase, and the optimum temperature of M. aeruginosa growth was 20–25 °C. Additionally, TEPS and microcystins (MCs) secretion peaked at 20–25 °C. The cell density accumulated rapidly from 26 °C to 35 °C. Furthermore, enzymes of RuBisCO and FBA related to photosynthetic activity were confirmed to contribute to the metabolism, as well as mcyB gene was affected by elevated temperature. Our results provide insights of the physiological effects and metabolic activity during annual cycle of M. aeruginosa. And it is predicted that global warming may promote the earlier recruitment of M. aeruginosa, extend the optimum growth period, enhance the toxicity, and finally intensify M. aeruginosa blooms.

Promoting vivianite recovery: Crucial role of tightly-bound extracellular polymeric substances

Vivianite is an important bio-induced product of dissimilated iron reduction (DIR), which exists widely in activated sludge and digested sludge of wastewater treatment plants (WWTPs). Extracellular polymeric substances (EPS) surrounding dissimilatory iron-reducing bacteria (DIRB) were reported to be transient media for microbial extracellular electron transfer (EET) and DIR. However, what happens when a layer of EPS envelops microbial cells during vivianite recovery? This study tried to assess the relationship between iron reduction and electron transfer by stripping EPS layer-by-layer, then clarified the delamination of different EPS layers on the performance of vivianite recovery. After stripping total EPS (PC-EPS batches) of DIRB, the Fe (III) reduction efficiencies (RFe), Fe (III) reduction rate (VFe), and vivianite formation efficiency (RV) decreased by 33%, 37%, and 30%, respectively. Tightly bound-EPS (TB-EPS) was confirmed to play a crucial role in vivianite formation, which contributed 28–51% to vivianite recovery and 34–55% to iron reduction. Electronic shuttle substances were contained in TB-EPS, which enhanced the EET rate and efficiency between cells and Fe (III). The polysaccharide content of TB-EPS was positively related to adhesiveness between cells and mineral, as well as the iron reduction efficiency. Furthermore, redox proteins (PpcA, OmcB) were embedded in EPS, which shortened the distance of electron hopping to across the EPS layer and further accelerated vivianite recovery from wastewater. Long-term operation of anaerobic reactors confirmed that TB-EPS played critical roles in bio-induced vivianite formation.

Chemical extraction and quantification of extracellular polymeric substances in unspiked-metaldehyde and spiked-metaldehyde of rubber aerobic granular sludge and molasses aerobic granular sludge

The building blocks of biofilm are called extracellular polymeric substances (EPSs), and they are composed of biopolymers. The optimal properties of EPSs include chemical interactions, resistance to dehydration, and resilience in the face of hazardous shock loads. It is necessary to have an understanding of EPSs extraction, composition, and the biomatrix's reaction to contaminants. This research aims to conduct chemical extraction to determine the amounts of EPSs found in unspiked-metaldehyde and spiked-metaldehyde rubber aerobic granular sludge (&lt;i&gt;r&lt;/i&gt;AGS) and newly invented molasses aerobic granular sludge (&lt;i&gt;m&lt;/i&gt;AGS). The &lt;i&gt;r&lt;/i&gt;AGS was produced by feeding synthetic wastewater into sequencing batch reactors (SBRs). The &lt;i&gt;m&lt;/i&gt;AGS was developed by adding the &lt;i&gt;r&lt;/i&gt;AGS with different volumes of molasses wastewater. The EPSs were successfully extracted and identified by quantifying the number of proteins and carbohydrates present in each &lt;i&gt;r&lt;/i&gt;AGS and &lt;i&gt;m&lt;/i&gt;AGS condition. Hydroxyl, carboxyl, and amino groups showed that &lt;i&gt;r&lt;/i&gt;AGS and &lt;i&gt;m&lt;/i&gt;AGS EPSs contained carbohydrates and proteins. The compositions of the EPSs showed a strong correlation with the concentrations of metaldehyde in the spiked-metaldehyde &lt;i&gt;r&lt;/i&gt;AGS and &lt;i&gt;m&lt;/i&gt;AGS samples. When the total EPSs are taken into consideration, both &lt;i&gt;r&lt;/i&gt;AGS and &lt;i&gt;m&lt;/i&gt;AGS that are formed are effective treatments for metaldehyde treatment in wastewater.

Impacts and mechanism of total organic sulfur in extracellular polymeric substances on fouling of the anaerobic self-forming dynamic membrane bioreactor

A side-stream cross-flow anaerobic self-forming dynamic membrane bioreactor (AnSFDMBR) connected with an up-flow anaerobic sludge blanket (UASB) reactor was used to replace the UASB and middle settling tank in sulfate reduction, autotrophic denitrification, and nitrification integrated (SANI) process. To reduce the water and energy consumption for AnSFDMBR backwashing, the optimum condition of trans-membrane pressure (TMP)-based backwashing were investigated under sequential batch operations. Based on the optimum condition, a lab-scale AnSFDMBR was operated continuously for 243 days without a drop in flux, with good effluent quality. In the sequential batch operation study, AnSFDMBR showed no significant accumulation of total extracellular polymeric substances (EPSs) during 40 days of operation without backwashing. Polysaccharides slightly accumulated in the dynamic layer regardless of the final TMP and operation duration, indicating that TMP-based backwashing is suitable for the studied AnSFDMBR system. In long-term operation with TMP-based backwashing, total EPSs in dynamic layer to sludge ratio dramatically dropped during the fouling phase. According to the evaluation of total organic sulfur (TOS) in dynamic layer and mesh EPSs, it was found that TOS, mainly poly-sulfur generated in UASB reactor, played a critical role in fouling in saline wastewater treatment. TOS deposited on the mesh over 40% by weight of the total EPS in the fouling phase clearly led to system fouling. Additional sequential batch operations with different seawater percentages (%V/V) (control, 10%, 20%, 30% and 50%) demonstrated that the TOS accumulation on the mesh increased with time and salinity which also aligned with the long-term operation.

The mechanism of extracellular polymeric substances in the formation of activated sludge flocs

In this study, activated sludge (AS) presented high aggregation ability of 56.9% to form flocs. To elucidate the effects of extracellular polymeric substances (EPS) on the AS flocs formation, four EPS fractions, including soluble EPS (S-EPS), loosely bound EPS (LB-EPS), ionic tightly bound EPS (ionic TB-EPS) and hydrophobic tightly bound EPS (hydrophobic TB-EPS), were extracted and characterized. The ionic TB-EPS were the largest fraction, accounting for 51.3% of the total EPS. By extracting EPS fractions, it was shown that ionic TB-EPS followed by hydrophobic TB-EPS played positive contribution to the formation of AS flocs, while both S-EPS and LB-EPS had negative effects. Furthermore, chemical reagents (Urea, SDS and EDTA) assays, zeta potential and spectroscopic assays were conducted to explore the role of hydrogen bonds, electrostatic interaction and hydrophobic interaction in the formation of AS flocs. The result showed that ionic TB-EPS contained sufficient cation-binding sites, such as -OH and CO-O- groups, and promoted AS aggregation through ion bridging. Hydrophobic TB-EPS contained hydrophobic groups and promoted AS aggregation through hydrophobic action. A high proportion of uronic acids resulted in high surface negative charge, which might be the reason that LB-EPS and S-EPS inhibited AS aggregation through electrostatic repulsion. From the perspective of molecule structure, the proteins in ionic TB-EPS had a loose structure, which fully exposed the inner cation-binding sites. This might be the critical reason why ion bridging acted as the main force for the formation of AS flocs. This study provided a new sight for the extraction of EPS fractions and expanded the understanding of EPS in the formation of AS flocs.

Effect of different forms and components of EPS on sludge aggregation during granulation process of aerobic granular sludge.

The characteristics of three forms of extracellular polymeric substances (EPS) and their component proteins (PN) and polysaccharides (PS) during the granulation process of aerobic granular sludge (AGS) and their effects on the aggregation performance of AGS were studied. Meanwhile, Pearson correlation analysis was conducted to explore the correlations between different forms of EPS and their components and sludge properties in the granulation process. The results showed that the AGS reactor could be started within 40 days by using the strategy of decreasing sedimentation time, and the sludge granulation degree was 90.22%. It was found that the content of tightly bound EPS (TB-EPS) accounted for 72.81-93.11% of the total EPS. Compared with dissolved EPS (S-EPS) and loosely bound EPS (LB-EPS), TB-EPS had a great impact on the zeta potential and hydrophobicity of sludge surface. During the granulation process, the content and relative hydrophobicity of PN in TB-EPS increased from 30.38±0.12mg/g VSS to 83.99±2.05mg/g VSS and 37.63%-71.25%, respectively. Through the flocculation experiment, it showed that the contribution rate of TB-EPS to the flocculation capacity of sludge cells increased from 13.30% to 35.37% and that of PN to flocculation capacity was 76.93%, which was 6.8 times that of PS. It indicated that TB-EPS played the most critical role in the formation of AGS and the PN in EPS was the major contributor to promoting the formation of AGS and enhance its aggregation performance. This research could further understand the role of various forms of EPS and its components in the process of sludge granulation, reveal the sludge granulation mechanism, and provide theoretical support for the rapid start-up of AGS reactor.

Performance and mechanism of Chlorella in swine wastewater treatment: Roles of nitrogen-phosphorus ratio adjustment and indigenous bacteria

The effects of adjusting the nitrogen-phosphorus (N/P) ratio of wastewater and indigenous bacteria on swine wastewater treatment by Chlorella sp. HL were investigated. The optimal N/P ratio of Chlorella in swine wastewater was 20 by adjusting the phosphorus concentration. The participation of indigenous bacteria increased total extracellular polymeric substances content, which was beneficial to maintain the stability of the algal-bacterial consortium, and improved the algal density and the removal rate of total nitrogen, total phosphorus, and chemical oxygen demand by 47.8%, 24.0%, 30.7%, and 326.7%, respectively. Proteobacteria was the dominant phylum with the relative abundance of 71.58% in the algal-bacterial system at optimal N/P ratio, and Brevundimonas, Chryseobacterium, and Pseudomonas played positive roles in the establishment of symbiotic systems at the genus level. These results provide a theoretical basis for the construction of an efficient algal-bacterial symbiotic system in swine wastewater treatment and support for commercial scale-up.