Proteins In Extracellular Polymeric Substances Research Articles

Biofilm formation and extracellular polymeric substance (EPS) production by Bacillus haynesii and influence of hexavalent chromium

Biofilm-forming bacteria play a key role in the removal of heavy metals including hexavalent chromium [Cr(VI)] from contaminated sites. In this study, biofilm-forming B. haynesii was examined for extracellular polymeric substances (EPS) production and hexavalent chromium [Cr(VI)] reduction potential. Exposure of B. haynesii with Cr(VI) (12.5–100 mg L-1) for 48 h enhanced pellicle dry weight (20–24%), cell-size (5.1–23.2%) and cell granularity (8.5–19.2%). Also, EPS production was increased by 10–35% by promoting the synthesis of protein (94–119%) and polysaccharide (2–33%) components in EPS. Further, the reduction (27.7 %) and distribution (15.87%) of Cr(VI) were mainly mediated by EPS than the other cellular fractions. Findings of the study suggest that the EPS from B. haynesii was efficiently reduced to Cr(VI) present in aqueous medium and the potential of the organism can be further explored for the mitigation of Cr(VI) contamination.

Biochar assisted cellulose anaerobic digestion under the inhibition of dodecyl dimethyl benzyl ammonium chloride: Dose-response kinetic assays, performance variation, potential promotion mechanisms

This study evaluated the inhibitory effect and mitigation strategy of dodecyl dimethyl benzyl ammonium chloride (DDBAC) suppression on anaerobic digestion. With the 12 h-suppression, only 16.64% of anaerobes were alive, and acetotrophic methanogens were significantly inhibited. As for batch test, DDBAC suppression significantly prolonged the start-up of systems and decreased the biogas production. In cellulose semi-continuous digestion process, the DDBAC suppression induced volatile fatty acids accumulation and pH decrease. However, the biochar amended reactor effectively mitigated the DDBAC suppression and achieved 370.5 mL/d·g-chemical-oxygen-demand biogas production. Moreover, 17.8% more protein in extracellular polymeric substances was secreted as the bio-barrier to defense the DDBAC suppression. Furthermore, microbial analysis showed that biochar addition selectively enriched directed interspecies electron transfer (DIET) participant bacteria (Anaerolineaceae and Syntrophomonas) and methanogens (Methanosaeta and Methanobacterium). Meanwhile, the potential metabolic pathway analysis showed that the abundance of amino acids and energy metabolism were increased 28% and 8%, respectively. The abundance of encoding enzyme related to hydrogenotrophic and acetotrophic methanogenesis enriched 1.88 times and 1.48 times, respectively. These results showed the performance and mechanisms involved in DIET establishment with ethanol stimulation biochar addition.

Steel pickling rinse wastewater treatment by two-stage MABR system: Reactor performance, extracellular polymeric substances (EPS) and microbial community

A bench-scale two-stage membrane-aerated biofilm reactor (MABR) system was applied to treat steel pickling rinse wastewater with high salinity and refractory organic. The effects of salinity and aeration pressure on the treatment efficiency, extracellular polymeric substances (EPS) characteristics and microbial community structure were studied. The optimal removal efficiencies of COD, NH+ 4-N and TN reached to 62.84%, 99.57% and 51.65%, respectively. Shortcut nitrification was achieved at low aeration, and the salinity less than 4% did not remarkable affect system performance. Colorimetric determination, three-dimensional exaction-emission matrix (3D-EEM) and Fourier transform infrared spectrum (FTIR) were employed to characterize the content and composition of proteins (PN) and polysaccharides (PS) in EPS of the biofilm. The results indicated that PN, not PS, response to changes of environmental conditions played a key role. Moreover, EPS might alleviate intracellular and extracellular osmotic pressure imbalance induced by high salinity, which imparted the biofilm in MABR with prominent salt-tolerant. High-throughput sequencing displayed that nitrifiers (Nitrosomonas, Nitrospira), denitrifiers (Dechloromonas, Hyphomicrobium, Denitromonas, Denitratisoma, Candidatus_Competibacter) and aerobic denitrifiers (Pseudomonas, Thauera) were predominant salt-tolerant bacteria.

Inhibitory activities of Typhonium trilobatum (L.) Schott on virulence potential of multi-drug resistant toxigenic Vibrio cholerae

Cholera is a serious epidemic disease caused by the toxigenic strains of Vibrio cholerae belonged to O1 or O139 serogroups. The emergence of antibacterial resistance in V. cholerae is an increasing concern. Natural product drug invention and Ethnopharmacology may demonstrate a considerable expectation under this circumstance. Traditionally, leaves of Typhonium trilobatum (L.) Schott (locally known as Ghatkanchu or Bengal Arum) are employed for treatment of gastrointestinal disorder in different region of India.The objective of the present study was to evaluate the antibacterial, and antibiofilm activities of methanol extract of T. trilobatum leaves (METTL) against the strains of multi-drug resistant (MDR) Vibrio cholerae (serotypes O1, O139, non-O1, and non-O139) which are responsible for watery diarrhea such as cholera.MIC, MBC and time-kill kinetic studies were used for evaluation of In vitro antibacterial activity of METTL. Microdilution method and Confocal laser scanning microscopy were used to evaluate biofilm-inhibitory activities. The gene expression was analyzed by performing Quantitative real-time PCR (qRT-PCR).METTL showed antibacterial activity with MIC and MBC at 1–32 mg/mL and 8–32 mg/mL, respectively against the clinical strains of Vibrio cholerae belonged to different serogroups. METTL showed significant (P < 0.05) inhibitory activity on the formation of biofilm by V. cholerae SG24, with 81.3, 75.8, and 69.6% of inhibition at MIC, ½ MIC and ¼ MIC, respectively. METTL showed also significant (P < 0.05) inhibitory activity on the formation of extracellular polymeric substances (EPS) formation by V. cholerae SG24, with 89.41, and 99.26% of inhibition of EPS protein and EPS carbohydrate at MIC, respectively.METTL significantly (p < 0.01) inhibited the Cholera toxin (CT) production by the V. cholerae strain SG24 evaluated by the CT - ELISA assay. The cholera toxin production was reduced by 76.26%, 48.76% and 29.93 at MIC (8 mg/mL), ½ MIC (4 mg/mL) and ¼ MIC (2 mg/mL), respectively. METTL was shown to repress ctxAB gene transcription 1.76 fold (p < 0.05) at sub-bactericidal concentration (¼ MIC). We also found that the expression of cholera toxin activator genes, toxT and tcpP was reduced by 11.56- fold (p < 0.001) and 23.52- fold (p < 0.001), respectively, at sub-bactericidal concentration (¼ MIC). Transcription of the following genes was repressed: vpsR (1.8-fold; p < 0.05), Bap1 (1.53-fold; p ≤ 0.05), and rmbA (2.89-fold) by METTL at sub-bactericidal concentration. The expression of vpsT was also repressed by 1.5-fold (p < 0.01) at sub-bactericidal concentration.The active Typhonium trilobatum (L.) leaves extract may be suggested as an substitute for the treatment of MDR V. cholerae infection and could be used as prospective source for the development of novel antimicrobial compound/s and biofilm-inhibitory drug/s useful for the treatment of cholera and diarrheal patients. The results obtained here also validate scientifically the traditional uses of Typhonium trilobatum (L.) in India employed for the treatment of gastrointestinal disorder. Further studies should be directed at purifying and characterizing these antibacterial principles against Vibrio cholerae.

Insight into the role of different extracellular polymeric substances components on trimethoprim adsorption by activated sludge

Adsorption is the primary mechanism of antibiotic removal in wastewater treatment plants, wherein the extracellular polymeric substances (EPS) of the activated sludge play an important role. Due to their complex characteristics, the effect of EPS components on antibiotic adsorption is unknown. Therefore, in this study, the role of main components of EPS in antibiotic adsorption was explored using enzymatic treatment. The results revealed that proteinase K and α-amylase can efficiently hydrolyse the proteins and polysaccharides of EPS. The protein content of EPS reduced from 31.25 mg/g VSS to 21.53, 18.75, and 10.76 mg/g VSS, after treatment with proteinase K, α-amylase and their combination, respectively; the polysaccharides content also observed a similar decrease from 15.20 mg/g VSS to 8.22, 7.83, and 6.03 mg/g VSS, respectively. The humic substance in EPS was stable during enzymatic treatment. The equilibrium adsorption capacity of activated sludge treated by enzyme for trimethoprim (TMP)- a typical antibiotic, was significantly increased from 2.19 μg/g VSS to 4.68, 5.34, and 8.36 μg/g VSS after treatment with proteinase K, α-amylase and their mixture. The adsorption process was adequately described by pseudo-second-order kinetic model. A multivariable linear regression model was subsequently used to quantify the adsorption capacity of activated sludge for TMP considering the concentration of EPS components. The modelling and validated results showed that the model could satisfactorily predict the TMP adsorption capacity. The results of this study can provide new insights into the role of EPS on antibiotic transformation in biological wastewater treatment systems.

Effects of Waste Activated Sludge Extracellular Polymeric Substances on Biosorption

Extracellular polymeric substances (EPS) reportedly make up approximately half of the organic matter in activated sludge (AS), and therefore strongly influence AS properties. This study evaluated the component fractions of EPS normalized to volatile suspended solids (VSS) in waste activated sludge (WAS) from a trickling-filter-solids contact process (TF/SC) and its ability to biosorb organic matter from raw wastewater with 30 min of contact time. Biosorption is the process in which organic matter (carbohydrates, proteins, humic acids, DNA, uronic acids, and lipids) in a sorbate, such as raw wastewater, sorbs onto a sorbent such as WAS. A statistically significant correlation was found between both the total concentration of EPS and the proteins component of the EPS and the biosorption removal of soluble chemical oxygen demand (sCOD) and truly soluble COD (ffCOD). Thus, the biosorption of soluble forms of COD can accurately be predicted by quantifying just the amount of proteins in WAS-associated EPS. No significant correlations were found for the biosorption of colloidal COD (cCOD). WAS biosorbed 45–75 mg L−1 of COD in 30 min. WAS absorbed or stored the proteins fraction of the soluble microbial products (SMP) during the biosorption process. Higher concentrations of humic acids were found in the biosorption process effluent than in the untreated wastewater, which warrants further study. Longer cation exchange resin (CER) extraction times yielded more total EPS from the sludge: 90 ± 9, 158 ± 3, and 316 ± 44 mg g−1 VSS, for 45-min, 4-h, and 24-h extraction times, respectively. Thus, EPS extracted represented only 9%, 15.8%, and 31.6% of the VSS, respectively, raising questions about whether the accurate characterization of EPS can be performed using the typical extraction time of 45 min due to different extraction rates for different components. It was found that the humic acids fraction was extracted much more slowly than the other fractions.

Contribution, Composition, and Structure of EPS by In Vivo Exposure to Elucidate the Mechanisms of Nanoparticle-Enhanced Bioremediation to Metals.

Bacterial extracellular polymeric substances (EPS) have been recently found to contribute most for metal removal in nanoenhanced bioremediation. However, the mechanism by which NPs affect EPS-metal interactions is not fully known. Here, Halomonas sp. was employed to explore the role of EPS after in vivo exposure to Cd/Pb and polyvinylpyrrolidone (PVP) coated iron oxide nanoparticles (IONPs, 20 mg L-1) for 72 h. Cd-IONPs produced the highest concentrations of EPS proteins (136.3 mg L-1), while Cd induced the most production of polysaccharides (241.0 mg L-1). IONPs increased protein/polysaccharides ratio from 0.2 (Cd) to 1.2 (Cd-IONPs). The increased protein favors the formation of protein coronas on IONPs surface, which would promote Cd adsorption during NP-metal-EPS interaction. FTIR analysis indicated that the coexistence of Cd and IONPs interacted with proteins more strongly than with polysaccharides. Glycosyl monomer analyses suggested mannose and glucose as target sugars for EPS complexation with metals, and IONPs reduced metal-induced changes in monosaccharide profiles. Protein secondary structures changed in all treatments, but we could not distinguish stresses induced by metals from those by IONPs. These findings provide greater understanding of the role of EPS in NP-metal-EPS interaction, providing a better underpinning knowledge for the application of NP-enhanced bioremediation.

Enhancing the formation of simultaneous nitrification and denitrification (SND) biofilm and nitrogen removal performance using two-units IFFAS process filled with surface-modified carriers

Moving-bed biofilm reactor (MBBR) and integrated floating-film activated sludge (IFFAS) are considered as well-established technologies for simultaneous nitrification and denitrification (SND). However, SND biofilm formation and nutrient removal performance are generally limited by the innate physico-chemical properties of conventional suspended carriers and/or insufficient carbon source available for denitrification. In this study, novel surface-modified carriers were developed via introducing mixed functional materials (natural zeolite, cationic polyquaternium-10, insoluble biodegradable organic polybutylene succinate/bamboo powder) into polyethylene. The modified carrier revealed enhanced hydrophilicity, positive surface charge, higher surface free energy and carbon source slow release capacity. Biofilm growth feature showed that the surface-modification resulted in more biofilm formation and more polysaccharide and protein in extracellular polymeric substances (EPS), which was structural and functional beneficial for substrate capture, transfer and intercellular interactions between nitrifiers and denitrifiers. IFFAS reactor filled with novel modified carriers was upgraded by dividing into two vertical units, where alternate feeding and aeration was conducted to create an anoxic unit to enhance the utilization of influent carbon source for denitrification as well as an aerobic unit to strengthen nitrification. The two-units IFFAS reactor exhibited highly efficient SND performance with TN removal efficiency of 59.9–65.9%, increased by 12.3–13.5% compared with control group. This study proposed an approach towards enhancement of functional biofilm formation and SND performance, and had fascinating potential in actual applications.

Effect of different D-amino acids on biofilm formation of mixed microorganisms.

This study aimed to determine the effects of D-tyrosine, D-aspartic acid, D-tryptophan and D-leucine on biofilm formation of mixed microorganisms. Results showed that, in the attachment stage, D-amino acids caused significant reduction in adhesion efficiency of mixed microorganisms to the membrane surface. Moreover, D-amino acids have a promoting effect on the reversible adhesion of mixed microorganisms. The addition of D-amino acid generally inhibited the biofilm biomass, of which D-tyrosine has the best inhibition effect. With the effect of D-tyrosine, D-aspartic acid, D-tryptophan and D-leucine, the protein in extracellular polymeric substance (EPS) decreased by 8.21%, 7.65%, 3.51% and 11.31%, respectively. The carbohydrates in EPS decreased by 29.53%, 21.44%, 14.60% and 10.54%, respectively. The results of excitation-emission matrix spectra (EEMs) suggested that the structural properties of the tyrosine-like proteins, tryptophan-like protein and humic-like acid might have changed by the D-amino acids.

Temperature-Dependent Influence of FliA Overexpression on PHL628 E. coli Biofilm Growth and Composition.

Biofilm growth and survival pose a problem in both medical and industrial fields. Bacteria in biofilms are more tolerant to antibiotic treatment due to the inability of antibiotics to permeate to the bottom layers of cells in a biofilm and the creation of altered microenvironments of bacteria deep within the biofilm. Despite the abundance of information we have about E. coli biofilm growth and maturation, we are still learning how manipulating different signaling pathways influences the formation and fitness of biofilm. Understanding the impact of signaling pathways on biofilm formation may narrow the search for novel small molecule inhibitors or activators that affect biofilm production and stability. Here, we study the influence of the minor sigma transcription factor FliA (RpoF, sigma-28), which controls late-stage flagellar assembly and chemotaxis, on biofilm production and composition at various temperatures in the E. coli strain PHL628, which abundantly produces the extracellular structural protein curli. We examined FliA’s influence on external cellular structures like curli and flagella and the biomolecular composition of the biofilm’s extracellular polymeric substance (EPS) using biochemical assays, immunoblotting, and confocal laser scanning microscopy (CLSM). At 37°C, FliA overexpression results in the dramatic growth of biofilm in polystyrene plates and more modest yet significant biofilm growth on silica slides. We observed no significant differences in curli concentration and carbohydrate concentration in the EPS with FliA overexpression. Still, we did see significant changes in the abundance of EPS protein using CLSM at higher growth temperatures. We also noticed increased flagellin concentration, a major structural protein in flagella, occurred with FliA overexpression, specifically in planktonic cultures. These experiments have aided in narrowing our focus to FliA’s role in changing the protein composition of the EPS, which we will examine in future endeavors.

Nitrogen removal, sludge activity and bacterial community in a sequencing batch reactor at different ciprofloxacin and ibuprofen concentrations

Nitrogen removal, sludge activity and bacterial community were examined at different ciprofloxacin and ibuprofen concentrations in a sequencing batch reactor. Ammonia nitrogen (NH 4 + -N) and chemical oxygen demand (COD) elimination rates decreased as the ciprofloxacin (belonged to antibiotics) and/or ibuprofen (belonged to nonsteroidal anti-inflammatory drugs) addition. Ciprofloxacin and ibuprofen had antagonistic inhibitions on the specific rates of nitrate reduction, nitrite oxidation and ammonium oxidation. The levels of protein and polysaccharide and the protein/polysaccharide ratio in extracellular polymeric substances rose as the increment of ibuprofen, ciprofloxacin and mixed ibuprofen/ciprofloxacin concentrations. Mixed ibuprofen/ciprofloxacin did not change the role of protein in extracellular polymeric substances in the bacterial defense to ibuprofen or ciprofloxacin, and tryptophan-like protein was the main protective response of bacteria to ibuprofen, ciprofloxacin and mixed ibuprofen/ciprofloxacin compared with aromatic protein. Both of nitrifiers and denitrifiers relative abundances were lower at the mixed ibuprofen/ciprofloxacin than at the alone ibuprofen stress, while were higher than at the single ciprofloxacin pressure. • Joint IBP and CIP had antagonistic impacts on SAOR, SNOR and SNRR. • Tryptophan-like PN in EPS was more important protective roles than aromatic PN. • Nitrifiers had a better tolerance to IBP, CIP and mixed IBP/CIP than denitrifiers.

Role of extracellular polymeric substances in the immobilization of hexavalent chromium by Shewanella putrefaciens CN32 unsaturated biofilms

Microbial remediation provides a promising avenue for the management and restoration of heavy metal-contaminated soils. Microorganisms in soils usually exist within unsaturated biofilms, however, their response to heavy metals is still limited compared to saturated biofilms. This work investigated the Cr(VI) immobilization by Shewanella putrefaciens CN32 unsaturated biofilms, and explored the underlying mechanisms of Cr(VI) complexation. Results reveal a dose-dependent toxicity of Cr(VI) to the growth of the unsaturated biofilms. During the early growth stage, the Cr(VI) addition stimulated more extracellular polymeric substances (EPS) production. In the meantime, the EPS were demonstrated to be the primary components for Cr(VI) immobilization, which accounted for more than 60% of the total adsorbed Cr(VI). The Fourier transform infrared spectra and X-ray photoelectron spectra corroborated that the binding sites for immobilizing Cr(VI) were hydroxyl, carboxyl, phosphoryl and amino functional groups of the proteins and polysaccharides in EPS. However, for the starved unsaturated biofilms, EPS were depleted and the EPS-bound Cr(VI) were released, which caused approximately 60% of the adsorbed Cr(VI) onto cell components and further aggravated the Cr(VI) stress to cells. This work extends our understanding about the Cr(VI) immobilization by unsaturated biofilms, and provides useful information for remediation of heavy metal-contaminated soils.

The hydrolysis and reduction of mixing primary sludge and secondary sludge with thermophilic bacteria pretreatment

Hydrolysis is widely used in sludge treatment as it enables to improve the biodegradability of sludge. However, there was little information about the hydrolysis efficiency with the mixture of primary and secondary sludge. In this study, the hydrolysis and reduction of mixed sludge after thermophilic bacteria (TB) pretreatment was investigated. Volume ratio of 1:3 (primary sludge: secondary sludge) had best release rate of organics, and the soluble chemical oxygen demand (SCOD), carbohydrate and protein in extracellular polymeric substances (EPS) decreased by 7.9, 4.1, 7.2 times, respectively. Meanwhile, the accumulation of SCOD, carbohydrate and protein in dissolved organic matter (DOM) was enhanced with mixed sludge. Excitation-emission matrix (EEM) spectroscopy proved that mixture of sludge was beneficial for the releasing of biodegradable substances. The sludge reduction of mixed sludge was higher than that of single sludge, and mixed sludge (1:3) had the best suspended solid (SS) and volatile suspended solids (VSS) reduction rate of 27.2% and 34.3%.

Effect of biofilm media application on biomass characteristics and membrane permeability in the biological spatiotemporal phase-separation process

In this study, the effect of moving-bed biofilm carrier, a membrane, and simultaneous employment to solve the settleability problem derived from readily biodegradable organics application in the biological spatiotemporal phase separation and variation of biomass characteristics by biofilm media application were investigated. No significant differences of the treatability among each case were observed. Although the moving-bed biofilm process is commonly used for the treatment of organics and nitrogen in wastewater, phosphorus was also removed perfectly owing to the detached biomass containing orthophosphate was discarded sufficiently. Additionally, the concentration of the suspended biomass decreased significantly, and the settled volume index was improved dramatically from 700 to 100 mL/g after the addition of the biofilm carrier. Although the extracellular polymeric substance (EPS) concentration increased with media addition, the back-flushing interval of the membrane was lengthened. The increment of polysaccharide in the EPS and the non-existence of protein in the soluble EPS were clearly observed. Thus, the suspended-biomass decrement due to media addition and the protein concentration in the EPS played important roles for fouling control. This study provides valuable information on state alternation of biomass by biofilm media application on biological suspended growth wastewater treatment process.

Impact of algal extracellular polymeric substances on the environmental fate and risk of molybdenum disulfide in aqueous media

Molybdenum disulfide (MoS2) poses great potential in water treatment as a popular transition metal dichalcogenide, arousing considerable concern regarding its fates and risk in aquatic environments. This study revealed that the interplay with extracellular polymeric substances (EPS) of freshwater algae significantly changed the properties and toxicity of MoS2 to aquatic fish. The predominant binding of aromatic compounds, polysaccharides, and carboxyl-rich proteins in EPS on the 1T polymorph of MoS2 via hydrophilic effects and the preferential adsorption of carboxylic groups contributed to morphological alterations, structural disorders (band gap and phase alterations), and the attenuated aggregation of MoS2 in aqueous solutions. Electron charge transfer and n-π* interactions with EPS decreased the catalytic activity of MoS2 by inhibiting its capability of generating reactive intermediates. The dissolution of MoS2 slowed down after interacting with EPS (from 0.089 to 0.045 mg/L per day) owing to rapid initial oxidation (i.e., forming Mo-O bond) and carbon grafting. Notably, the morphological and structural alterations after EPS binding alleviated the toxicity (e.g., malformation and oxidative stress) of MoS2 to infantile zebrafish. Our findings provide insights into the environmental fate and risk of MoS2 by ubiquitous EPS in natural waters, serving as valuable information while developing water treatment processes accordingly.

Effects of different exogenous cadmium compounds on the chemical composition and adsorption properties of two gram-negative bacterial EPS

Three different Cd(II) compounds were used to regulate Pseudomonas aeruginosa and Alcaligenes faecalis EPS (extracellular polymeric substances). The purpose of this study was to improve the content of EPS protein and the adsorption capacity of Cd(II) by different Cd(II) compounds. The results showed that Cd(NO3)2 had the best stress/induction effect on the two strains. Under the best stress/induction, the protein in EPS of the two strains increased most obviously, and the adsorption capacity of Cd(II) was increased by more than 40%. Under these conditions, the kinetics of the adsorption process of Cd(II) by Cd(NO3)2-EPSA. F (EPS produced by Alcaligenes faecalis under Cd(NO3)2 stress) could be well fitted by the Langmuir isotherm model, and the theoretical maximum adsorption amount of 1111.11 mg/g EPS could be obtained. The results of 3D-EEM, FTIR and XPS indicated that proteins, especially CO, CN and NH in proteins, played a major role in the removal of Cd(II) by Cd(NO3)2-EPSA. F. The results of this study show that the addition of Cd(NO3)2 can effectively regulate the content of chemical components, especially the content of protein, and thus greatly improve the removal efficiency of heavy metals, which shows great application prospects in the prevention and control of heavy metal pollution.

Cultivation of sulfide-driven partial denitrification granules for efficient nitrite generation from nitrate-sulfide-laden wastewater

Sulfide partial denitrification (SPD) is an alternative pathway for nitrite production accompanied with elemental sulfur (S0) production for nitrate removal from wastewater with anammox. In this study, the SPD granular sludge was cultivated for the first time in an upflow anaerobic sludge bed (UASB) reactor to reach the efficacy of maximum nitrate-to-nitrite transformation ratio of 92% and an in-situ maximum NO3−-N reduction rate of 2.46 kg-N/m3-d, both much higher than literature results. Mature granules had an average particle size of 2.52 mm and hold smooth surface with excess rod bacteria. The elements Ca and S, and proteins in extracellular polymeric substances contributed to granule structure's stability. Enriched Thiobacillus genus was proposed to accumulate nitrite at moderate HRT (2–6 h). The immobilized functional strains assist efficient partial nitrification reactions to be realized with formed S0 as byproduct.

Periodic chemical cleaning with urea: disintegration of biofilms and reduction of key biofilm-forming bacteria from reverse osmosis membranes

Biofouling is one of the major factors causing decline in membrane performance in reverse osmosis (RO) plants, and perhaps the biggest hurdle of membrane technology. Chemical cleaning is periodically carried out at RO membrane installations aiming to restore membrane performance. Typical cleaning agents used in the water treatment industry include sodium hydroxide (NaOH) and hydrochloric acid (HCl) in sequence. Rapid biofilm regrowth and related membrane performance decline after conventional chemical cleaning is a routinely observed phenomenon due to the inefficient removal of biomass from membrane modules. Since extracellular polymeric substances (EPS) make up the strongest and predominant structural framework of biofilms, disintegration of the EPS matrix should be the main target for enhanced biomass removal. Previously, we demonstrated at lab-scale the use of concentrated urea as a chemical cleaning agent for RO membrane systems. The protein denaturation property of urea was exploited to solubilize the proteinaceous foulants, weakening the EPS layer, resulting in enhanced biomass solubilization and removal from RO membrane systems. In this work, we investigated the impact of repeated chemical cleaning cycles with urea/HCl as well as NaOH/HCl on biomass removal and the potential adaptation of the biofilm microbial community. Chemical cleaning with urea/HCl was consistently more effective than NaOH/HCl cleaning over 6 cleaning and regrowth cycles. At the end of the 6 cleaning cycles, the percent reduction was 35% and 41% in feed channel pressure drop, 50% and 70% in total organic carbon, 30% and 40% in EPS proteins, and 40% and 66% in the peak intensities of protein-like matter, after NaOH/HCl cleaning and Urea/HCl cleaning, respectively. 16S ribosomal RNA (rRNA) gene sequencing of the biofilm microbial community revealed that urea cleaning does not select for key biofouling families such as Sphingomonadaceae and Xanthomonadaceae that are known to survive conventional chemical cleaning and produce adhesive EPS. This study reaffirmed that urea possesses all the desirable properties of a chemical cleaning agent, i.e., it dissolves the existing fouling layer, delays fresh fouling accumulation by inhibiting the production of a more viscous EPS, does not cause damage to the membranes, is chemically stable, and environmentally friendly as it can be recycled for cleaning.

Enhanced Secretions of Algal Cell-Adhesion Molecules and Metal Ion-Binding Exoproteins Promote Self-Flocculation of Chlorella sp. Cultivated in Municipal Wastewater.

The mechanism of self-flocculation remains unclear, partially impeding its efficiency enhancement and commercial application of microalgae-based municipal wastewater (MW) bioremediation technology. This study revealed the contributions of exoproteins [PN, proteins in extracellular polymeric substances (EPS)] to the separation of indigenous microalgae from treated MW. Compared to the low light intensity group, the high light intensity (HL) group produced Chlorella sp. with 4.3-fold higher self-flocculation efficiencies (SE). This was attributed to the enriched biological functions and positional rearrangement of increased PN within 2.9-fold higher EPS. Specifically, a total of 75 PN was over-expressed in the HL group among the 129 PN identified through label-free proteomics. The algal cell-adhesion molecules (Algal-CAMs) and metal-ion-binding PN were demonstrated as two dominant contributors promoting cell adhesion and bridging, through function prediction based on the contained domains. The modeled 3D structure showed that Algal-CAMs presented less hydrophilic α-helix abundance and were distributed in the outermost position of the EPS matrix, further facilitating microalgal separation. Moreover, the 10.1% lower hydrophily degree value, negative interfacial free energy (-19.5 mJ/m2), and 6.8-fold lower energy barrier between cells also supported the observed higher SE. This finding is expected to further fill the knowledge gap of the role of PN in microalgal self-flocculation and promote the development of biomass recovery from the microalgae-wastewater system.

Response of extracellular polymeric substances and microbial community structures on resistance genes expression in wastewater treatment containing copper oxide nanoparticles and humic acid

The extracellular polymeric substances (EPS) and microbial community structures were investigated in wastewater treatment containing copper oxide nanoparticles (CuO NPs) (reactor R1) and CuO NPs and humic acid (HA) (reactor R2) using both sequencing batch bioreactors (SBRs), and their response on resistance genes expression was analyzed. The removal of influent chemical oxygen demands (COD) and NH4+-N was moderately influenced under CuO NPs (5 mg/L) stress, while the function of HA (10 mg/L) was not reflected. However, the EPS production and microbial community were affected by the HA addition. The expression of different antibiotic resistance genes (ARGs), metal-resistance genes (MRGs), and intI1 was related to the primary compositions of polysaccharides and proteins in EPS and different microbial communities at the genus level. Furthermore, the expression of resistance genes was not stimulated under CuO NPs stress, and supplying HA was suggested to reduce their expression in wastewater treatment.