Composition Of Extracellular Polymeric Substances Research Articles

Integrated induction of silver resistance determinants and production of extracellular polymeric substances in Cupriavidus metallidurans BS1 in response to silver ions and silver nanoparticles

Although the antimicrobial mechanisms of nanomaterials have been extensively investigated, bacterial defense mechanisms associated with AgNPs have not been fully elucidated. We here report that dissolved Ag+ (>0.05 μg·mL-1) displayed higher toxicity on cell growth of strain Cupriavidus metallidurans BS1 (GCA_003260185.2) in comparison to 2 and 20 nm AgNPs. The genes necessary for synthesis of distinct abundance and composition of extracellular polymeric substances (EPS) were induced in strain BS1 exposed to Ag stress. This resulted in 20.1% (Ag(I)-EPS) and 24.2% (2 nm AgNPs-EPS) of the C=O band integrated intensities being converted into C-OH/C-O-C group vibrations and the Ag-O bond was formed between EPS and 20 nm AgNPs. Meanwhile, the expression of primary resistance genes of the cus, sil and cup operon encoding HME-RND-driven efflux systems as well as a PIB1-type ATPase (CupA) were significantly induced after exposure to Ag(I), 2 and 20 nm AgNPs, respectively. Furthermore, distinct genes involved in biosynthesis pathways responsible for production of EPS were induced to relieve the toxicity of Ag(I), 2 nm and 20 nm AgNPs. This combined action is one potential reason why strain BS1 displayed distinct resistances in response to Ag(I) compared to 2 and 20 nm AgNPs. This work will help in understanding processes important in bacterial defensive mechanisms to AgNPs.

Coaggregation dynamics in drinking water biofilms and implications for chlorine disinfection

Biofilms in drinking water (DW) systems persistently challenge traditional disinfection methods due to intricate microbial interactions, with coaggregation playing a crucial role in forming multispecies biofilms. This study examined the implications of coaggregation on tolerance towards sodium hypochlorite (NaOCl) disinfection. Dual-species biofilms were formed for seven days on polyvinyl chloride coupons, comprising a strain of the emerging pathogen Stenotrophomonas maltophilia and the coaggregating strain Delftia acidovorans 005 P. For comparison, dual-species biofilms were also formed with a non-coaggregation strain (D. acidovorans 009 P). The minimum bactericidal concentration (MBC) for each planktonic strain varied (D. acidovorans: 1 mg/L, S. maltophilia: 1.5 mg/L) below the safe DW treatment limits. However, high NaOCl doses (10 ×MBC and 100 ×MBC,) showed low efficacy against dual-species biofilms, indicating significant biofilm tolerance to disinfection. Membrane damage occurred at sub-MBC without culturability loss, underscoring biofilm resilience. The biofilm analysis revealed a complex interplay between the composition of extracellular polymeric substances and the architecture, which was influenced by the presence of the coaggregating strain. Overall, coaggregation significantly influenced biofilm formation and resilience, impacting NaOCl disinfection. These findings underscore the challenges of microbial interactions in biofilms, emphasizing the need for improved disinfection strategies to control biofilms in drinking water systems.

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.

Crucial role of extracellular polymeric substances from anammox sludge in wastewater phosphorus recovery via magnesium phosphate crystallization

The magnesium phosphate (Mg-P) crystallization based on anaerobic ammonia oxidation is of interest for simultaneous nitrogen and phosphorus removal in wastewater treatment due to cost-effectiveness. Understanding the role of extracellular polymeric substances (EPS) in crystal growth is pivotal for bio-induced Mg-P crystallization. In this work, the effects of EPS in Mg-P crystallization were thoroughly investigated. Results indicated that EPS decreased the initial crystallization rate but slightly improved the final crystallization yield. The maximum removal efficiencies of Mg and P ions increased by 2.8 % and 3.1 %, respectively, when EPS concentration was 80 mg/L. Additionally, the presence of EPS enhanced Mg3(PO4)2·10 H2O production and facilitated the formation of larger rhombic plate-like structures crystals (maximum particle size increased by 215 %). Analysis of EPS composition changes during the crystallization process and characterization of the final crystals suggested that tryptophan-like proteins preferentially involved in the Mg-P crystallization process, while β-sheet proteins potentially influencing crystal morphology. Furthermore, the formation of phosphate ester groups, hydrogen bonding, and COOH-Mg2+ complexes were considered triggering factors for the interaction between EPS and Mg-P crystals. This study elucidated the underlying mechanism of EPS on Mg-P crystallization, further enhancing the understanding of the interaction between inorganic mineralization processes and microorganisms.

Sweet Secrets: Exploring Novel Glycans and Glycoconjugates in the Extracellular Polymeric Substances of "Candidatus Accumulibacter".

Biological wastewater treatment relies on microorganisms that grow as flocs, biofilms, or granules for efficient separation of biomass from cleaned water. This biofilm structure emerges from the interactions between microbes that produce, and are embedded in, extracellular polymeric substances (EPS). The true composition and structure of the EPS responsible for dense biofilm formation are still obscure. We conducted a bottom-up approach utilizing advanced glycomic techniques to explore the glycan diversity in the EPS from a highly enriched "Candidatus Accumulibacter" granular sludge. Rare novel sugar monomers such as N-Acetylquinovosamine (QuiNAc) and 2-O-Methylrhamnose (2-OMe-Rha) were identified to be present in the EPS of both enrichments. Further, a high diversity in the glycoprotein structures of said EPS was identified by means of lectin based microarrays. We explored the genetic potential of "Ca. Accumulibacter" high quality metagenome assembled genomes (MAGs) to showcase the shortcoming of top-down bioinformatics based approaches at predicting EPS composition and structure, especially when dealing with glycans and glycoconjugates. This work suggests that more bottom-up research is necessary to understand the composition and complex structure of EPS in biofilms since genome based inference cannot directly predict glycan structures and glycoconjugate diversity.

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.

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

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

Exploring the role of phosphorus in the clogging formation of subsurface flow constructed wetland by two-dimensional visualization technique

Phosphorus (P) is associated with microbial activity and can be retained in constructed wetland (CW) through adsorption and chemical precipitation. However, its potential impact on clogging remains poorly studied. In this study, a two-dimensional (2D) visualization technique was utilized to investigate the effect of P on clogging process inside subsurface flow constructed wetland (SSF-CW) during the operation. Tracer images analysis showed that the preferential flow pathway proportions of P-containing group (P-CW) and P-free group (C-CW) were 30.38 ± 1.07 % and 9.72 ± 0.47 %, respectively. The fractal dimension of preferential flow pathways in C-CW (1.57 ± 0.02) was lower than in P-CW (1.70 ± 0.03), indicating that the preferential flow pathways in P-CW exhibited more diversion and tortuosity. The removal efficiencies of both CWs exhibited an initial increase followed by a decrease, with P-CW generally demonstrating higher pollutant removal efficiencies. In P-CW, the primary pathway for P removal was chemical precipitation dominated by apatite P (CaP). Furthermore, the addition of P altered the composition of extracellular polymeric substances (EPS) and reduced the stability of microbial aggregates, resulting in higher effective porosity. This study provides new insights into the role of P in the clogging of SSF-CW and would contribute to optimizing CW clogging management.

Effect of long-term dosage of hydrazine on mainstream anammox process: Biofilm characteristics and microbial community

The impact of the long-term trace hydrazine (N2H4) exogenous supplementation on activity of the anaerobic ammonium oxidation (anammox) biofilm was investigated in a moving bed biofilm reactor (MBBR) for mainstream wastewater treatment. The results of this study demonstrated that the addition of 2–5 mg/L N2H4 enhanced anammox biofilm activity, as evidenced by the augmented nitrogen removal rate (NRR), which increased from 113.4 g/(m3·d) to 126.7 g/(m3·d) with the introduction of 2 mg/L N2H4. However, a higher concentration of N2H4 (10 mg/L) suppressed anammox activity, leading to a reduced NRR of 91.5 g/(m3·d). Bioindicators revealed that the long-term addition of 2 mg/L N2H4 fostered the accumulation of anammox bacteria (AnAOB) biomass, elevating the volatile suspended solids (VSS) content by 12%. Moreover, the structural composition of extracellular polymeric substances (EPS) within the biofilm was altered, resulting in enhanced biofilm strength within the reactor. The protective mechanism of the biofilm was activated, and EPS secretion was stimulated by the continuous N2H4 supplementation. The introduction of an excess dosage of N2H4 led to alterations in the microbial communities, ultimately resulting in a decline in the performance of the reactor. These findings collectively illustrate that N2H4, as an intermediate product, can effectively enhance anammox activity within the MBBR for mainstream wastewater treatment. This study contributes to the understanding of the optimization strategies for anammox processes in wastewater treatment systems.

Effect of microwaves combined with peracetic acid to improve the dewatering performance of residual sludge.

The activated sludge process plays a crucial role in modern wastewater treatment plants. During the treatment of daily sewage, a large amount of residual sludge is generated, which, if improperly managed, can pose burdens on the environment and human health. Additionally, the highly hydrated colloidal structure of biopolymers limits the rate and degree of dewatering, making mechanical dewatering challenging. This study investigates the impact and mechanism of microwave irradiation (MW) in conjunction with peracetic acid (PAA) on the dewatering efficiency of sludge. Sludge dewatering effectiveness was assessed through capillary suction time (CST) and specific resistance to filtration (SRF). Examination of the impact of MW-PAA treatment on sludge dewatering performance involved assessing the levels of extracellular polymeric substances (EPS), employing three-dimensional excitation-emission matrix (3D-EEM), Fourier transform-infrared spectroscopy (FT-IR), and scanning electron microscopy. Findings reveal that optimal dewatering performance, with respective reductions of 91.22% for SRF and 84.22% for CST, was attained under the following conditions: microwave power of 600 W, reaction time of 120s, and PAA dosage of 0.25g/g MLSS. Additionally, alterations in both sludge EPS composition and floc morphology pre- and post-MW-PAA treatment underwent examination. The findings demonstrate that microwaves additionally boost the breakdown of PAA into •OH radicals, suggesting a synergistic effect upon combining MW-PAA treatment. These pertinent research findings offer insights into employing MW-PAA technology for residual sludge treatment.

Extracellular Polymeric Substance Composition Effects on Biosorption for Primary Carbon Diversion

This paper characterizes the composition of extracellular polymeric substances (EPSs) in different types of activated sludge (AS) processes and analyzes the biosorption of soluble organics when waste AS (WAS) is mixed with raw wastewater for primary carbon diversion. The fraction of AS organics identified as EPSs was 26% in a membrane bioreactor (MBR), 54% in conventional AS (CAS), and 51% in a trickling filter/solids contact (TF/SC) process. EPSs were found to be approximately 15% carbohydrates, 40% proteins, 40% humics, and 5% uronics in CAS and MBR AS. Biosorption was not correlated to the organic portion (VSS) of the WAS; however, statistically significant correlations were found for the total amount of EPSs (for TF/SC and CAS) and the protein fraction (for TF/SC and MBR) in the VSS. EPSs from different types of AS biosorbed the same amount of soluble organics, removing 1.43 ± 0.15 (n = 16) mg of soluble chemical oxygen demand (sCOD), and 1.20 ± 0.18 (n = 16) mg of truly soluble COD (ffCOD), per mg of cation exchange resin (CER) total extracted EPSs. Utilizing multiple extraction methods in series (CER–base–sulfide) increased EPS extraction yields by nearly 100% relative to CER alone and indicated different EPS fractionization for CAS (a smaller fraction of carbohydrates and a larger fraction of humics).

Evaluation of autotrophic process influencing extracellular polymeric substances in aerobic membrane bioreactor with expanded ASM model

A mathematical model was developed to predict the formation of both the autotrophic and heterotrophic extracellular polymeric substances (EPS) in the aerobic membrane bioreactor (MBR). Batch experimental results and 45-day operation data on a pilot MBR at a sludge retention time (SRT) of 20 d were used to calibrate and validate the model. Simulated MBR setup results demonstrated the key role of the influent COD and NH4+-N in governing the composition of heterotrophic and autotrophic EPS in the MBR. These results also revealed that the autotrophic EPS process was non-ignorable in the system. According to the autotrophic EPS simulation in the MBR, the EPS yield increased with increasing influent COD/NH4+-N ratio towards a constant level. The EPS yield was significantly influenced by the SRT, attributed to the autotrophic process's impact on EPS. Simulation results revealed a slight increase in EPS yield with an SRT of up to 5 days, followed by a rapid decrease beyond that threshold.

Dissolved organic carbon spurs bacterial-algal competition and phosphorus-paucity adaptation: Boosting Microcystis' phosphorus uptake capacity

Dissolved organic carbon (DOC) can alter the availability of background nutrients by affecting the proliferation of heterotrophic bacteria, which exerts a notable influence on algal growth and metabolism. However, the mechanism of how allochthonous DOC (aDOC) precipitates shifts in bacterial-algal interactions and modulates the occurrence of cyanobacteria blooms remains inadequately elucidated. Therefore, this study investigated the relationship between bacteria and algae under aDOC stimulation. We found that excess aDOC triggered the breakdown and reestablishment of the equilibrium between Microcystis and heterotrophic bacteria. The rapid proliferation of heterotrophic bacteria led to a dramatic decrease in soluble phosphorus and thereby resulted in the inhibition of the Microcystis growth. When the available DOC was depleted, the rapid death of heterotrophic bacteria released large amounts of dissolved phosphorus, which provided sufficient nutrients for the recovery of Microcystis. Notably, Microcystis rejuvenated and showed higher cell density compared to the carbon-absent group. This phenomenon can be ascribed that Microcystis regulated the compositions of extracellular polymeric substances (EPS) and the expression of relevant proteins to adapt to a nutrient-limited environment. Using time of flight secondary ion mass spectrometry (TOF-SIM) and proteomic analysis, we observed an enhancement of the signal of organic matter and metal ions associated with P complexation in EPS. Moreover, Microcystis upregulated proteins related to organic phosphorus transformation to increase the availability of phosphorus in various forms. In summary, this study emphasized the role of DOC in algal blooms, revealing the underestimated enhancement of Microcystis nutrient utilization through DOC-induced heterotrophic competition and providing valuable insights into eutrophication management and control.

The potential effects of N-Acyl homoserine lactones on aerobic sludge granulation during phenolic wastewater treatment

The formation of aerobic granular sludge (AGS) is relatively difficult during the treatment of refractory wastewater, which generally shows small granular sizes and poor stability. The formation of AGS is regulated by N-Acyl homoserine lactones (AHLs)-mediated quorum sensing (QS). However, the potential role of AHLs in AGS formation under the toxic stress of refractory pollutants and the heterogeneity in the distribution and function of AHLs across different aggregates are not well understood. This study investigated the potential effects of AHLs on the formation of AGS during phenolic wastewater treatment. The distribution and succession of AHLs across varying granular sizes and development stages of AGS were investigated. Results showed that AGS was successfully formed in 13 days with an average granular size of 335 ± 39 μm and phenol removal efficiency of >99%. The levels of AHLs initially increased and then decreased. C4-HSL and 3-oxo-C10-HSL were enriched in large granules, suggesting they may play a pivotal role in regulating the concentration and composition of extracellular polymeric substances (EPS). The content of EPS constantly increased to 149.4 mg/gVSS, and protein (PN) was enriched in small and large granules. Luteococcus was the dominant genus constituting up to 62% after the granulation process, and exhibited a strong association with C4-HSL. AHLs might also regulate the bacterial community responsible for EPS production, and pollutant removal, and facilitate the proliferation of slow-growing microorganisms, thereby enhancing the formation of AGS. The synthesis and dynamics of AHLs were mainly governed by AHLs-producing bacterial strains of Rhodobacter and Pseudomonas, and AHLs-quenching strains of Flavobacterium and Comamonas. C4-HSL and 3-oxo-C10-HSL might be the major contributors to promoting sludge granulation under phenol stress and play critical roles in large granules. These findings enhance our understanding of the roles that AHLs play in sludge granulation under toxic conditions.

Mechanistic insight into the aggregation ability of anammox microorganisms: Roles of polarity, composition and molecular structure of extracellular polymeric substances

The chemical characteristics of extracellular polymeric substances (EPS) of anammox bacteria (AnAOB) play a crucial role in the rapid enrichment of AnAOB and the stable operation of wastewater anammox processes. To clarify the influential mechanisms of sludge EPS on AnAOB aggregation, multiple parameters, including the polarity distribution, composition, and molecular structure of EPS, were selected, and their quantitative relationship with AnAOB aggregation was analyzed. Compared to typical anaerobic sludge (anaerobic floc and granular sludge), the anammox sludge EPS exhibited higher levels of tryptophan-like substances (44.82–56.52 % vs. 2.57–39.81 %), polysaccharides (40.02–53.49 mg/g VSS vs. 30.22–41.69 mg/g VSS), and protein structural units including α-helices (20.70–23.98 % vs. 16.48–19.32 %), β-sheets (37.43–42.98 % vs. 25.78–36.72 %), and protonated nitrogen (Npr) (0.065–0.122 vs. 0.017–0.061). In contrast, it had lower contents of β-turns (20.95–27.39 % vs. 28.17–39.04 %). These biopolymers were found to originate from different genera of AnAOB. Specifically, the α-helix-rich proteins were mainly derived from Candidatus Kuenenia, whereas the extracellular proteins related to tryptophan and Npr were closely associated with Candidatus Brocadia. Critically, these EPS components could drive anammox aggregation through interactions. Substantial amounts of tryptophan-like substances facilitated the formation of β-sheet structures and the exposure of internal hydrophobic clusters, which benefited the anammox aggregation. Meanwhile, extracellular proteins with high Npr content played a pivotal role in the formation of mixed protein-polysaccharide gel networks with the electronegative regions of polysaccharides, which could be regarded as the key component in the maintenance of anammox sludge stability. These findings provide a comprehensive understanding of the multifaceted roles of EPS in driving anammox aggregation and offer valuable insights into the development of EPS regulation strategies aimed at optimizing the anammox process.

Extracellular polymeric substances in electroactive biofilms play a crucial role in improving the efficiency of microbial fuel and electrolysis cells.

In microbial electrochemical cells (MECs), electroactive microbial biofilms can transmit electrons from organic molecules to anodes. To further understand the production of anodic biofilms, it is essential to investigate the composition and distribution of extracellular polymeric substance (EPS) in the MECs. In this study, the structure of EPS was examined in microbial electrolysis cells from mixed cultures forming biofilm using carbon fiber fabric anode. EPS was extracted from the anode biofilm of microbial electrolysis cells inoculated with mixed microbial culture. The anode biofilm yielded 0.4mg of EPS, of which 51.2% was humic substance, 16.2% was protein, 12.6% was carbohydrates, and 20% consisted of undetermined substances. Using epifluorescence microscopy, the composition of bacterial cells and their location inside EPS were studied, and the distribution of microbial communities was compared based on current density results in the presence of various carbohydrates. On the electrode surface, bacteria and EPS gathered or overlapped in various locations can affect microbial electrochemical performance. Our findings showed that EPS formation in electroactive biofilms would be important for enhanced efficiency of electricity- or hydrogen-producing microbial electrolysis cells.

Harnessing the potential of ginkgo biloba extract: Boosting denitrification performance through accelerated electron transfer

Ginkgo biloba extract (GBE) had several effects on the human body as one of the widely used phytopharmaceuticals, but it had no application in microbial enhancement in the environmental field. The study focused on the impact of GBE on denitrification specifically under neutral conditions. At the identified optimal addition ratio of 2% (v/v), the system exhibited a noteworthy increase in nitrate reduction rate (NRR) by 56.34%, elevating from 0.71 to 1.11 mg-N/(L·h). Moreover, the extraction of microbial extracellular polymeric substance (EPS) at this ratio revealed changes in the composition of EPS, the electron exchange capacity (EEC) was enhanced from 87.16 to 140.4 μmol/(g C), and the transfer impedance was reduced within the EPS. The flavin, fulvic acid (FA), and humic acid (HA) provided a π-electron conjugated structure for the denitrification system, enhancing extracellular electron transfer (EET) by stimulating carbon source metabolism. GBE also improved electron transfer system activity (ETSA) from 0.025 to 0.071 μL O2/(g·min·prot) and the content of NADH enhanced by 22.90% while significantly reducing the activation energy (Ea) by 85.6% in the denitrification process. The synergy of improving both intracellular and extracellular electron transfer, along with the reduction of Ea, notably amplified the initiation and reduction rates of the denitrification process. Additionally, GBE demonstrated suitability for denitrification across various pH levels, enhancing microbial resilience in alkaline conditions and promoting survival and proliferation. Overall, these findings open the door to potential applications of GBE as a natural additive in the environmental field to improve the efficiency of denitrification processes, which are essential for nitrogen removal in various environmental contexts.

Adaptive mechanism of the marine bacterium Pseudomonas sihuiensis-BFB-6S towards pCO2 variation: Insights into synthesis of extracellular polymeric substances and physiochemical modulation

Marine bacteria can adapt to various extreme environments by the production of extracellular polymeric substances (EPS). Throughout this investigation, impact of variable pCO2 levels on the metabolic activity and physiochemical modulation in EPS matrix of marine bacterium Pseudomonas sihuiensis - BFB-6S was evaluated using a fluorescence microscope, excitation-emission matrix (EEM), 2D-Fourier transform infrared correlation spectroscopy (2D-ATR-FTIR-COS), FT-NMR and TGA-DSC. From the results at higher pCO2 levels, there was a substantial reduction in EPS production by 58–62.8 % (DW). In addition to the biochemical composition of EPS, reduction in carbohydrates (8.7–47.6 %), protein (7.1–91.5 %), and lipids (16.9–68.6 %) content were observed at higher pCO2 levels. Functional discrepancies of fluorophores (tyrosine and tryptophan-like) in EPS, speckled differently in response to variable pCO2. The 2D-ATR-FTIR-COS analysis revealed functional amides (CN, CC, CO bending, -NH bending in amines) of EPS were preferentially altered, which led to the domination of polysaccharides relevant functional groups at higher pCO2. 1H NMR analysis of EPS confirmed the absence of chemical signals from H-C-COOH of proteins, α, β anomeric protons, and acetyl group relevant region at higher pCO2 levels. These findings can contribute new insights into the influence of pCO2 on the adaptation of marine microbes in future ocean acidification scenarios.

Synthesizing cationic polymers and tuning their properties for microalgae harvesting

This study reports a facile technique to synthesize and tune the cationic polymer, poly(3-acrylamidopropyl)trimethylammonium chloride (PAPTAC), in terms of molecular weight and surface change for harvesting three microalgae species (Scenedesmus sp., P.purpureum, and C. vulgaris). The PAPTAC polymer was synthesised by UV-induced free-radical polymerisation. Polymer tuning was demonstrated by regulating the monomer concentration (60 to 360 mg/mL) and UV power (36 and 60 W) for polymerisation. The obtained PAPTAC polymer was evaluated for harvesting three different microalgae species and compared to a commercially available polymer. The highest flocculation efficiency for Scenedesmus sp. and P. purpureum was observed at a dosage of 25 mg-polymer/g of dry biomass by using PAPTAC-90, resulting in higher flocculation efficiency than the commercial polymer. Results in this study show evidence of effective neutralisation of the negative charge surface of microalgae cells by the produced cationic PAPTAC polymer and polymer bridging for effective flocculation. The obtained PAPTAC polymer was less effective for harvesting C. vulgaris, possibly due to other factors such as cell morphology and composition of extracellular polymeric substances of at the cell membrane that may also influence harvesting performance.

Interrelation between extracellular polymer substances (EPSs) and MPs in an MBR

Nowadays, microplastics (MPs), as ubiquitous pollutants in aquatic ecosystems, present a major emerging threat. Wastewater treatment plants (WWTPs) are considered a hotspot of MP contamination. In this context, membrane bioreactors (MBRs), which combine conventional activated sludge processes with filtration, serve as an effective technology to remove MPs from wastewater. One of the main factors that affect MBR performance is the amount and composition of extracellular polymeric substances (EPSs) in the mixed liquor (ML). In previous studies carried out at lab-scale with model MPs, it has been found that the presence of MPs in wastewater treatment processes affect the EPS characteristics. In this work, the inter-relationship between EPSs and MPs has been investigated in a MBR at an industrial scale. Samples of the ML were taken from a WWTP during a six-month period and EPSs and MPs were analysed. It was observed that the higher the concentration of MPs, the higher the amount of EPSs were secreted by microorganisms, which seems to indicate that MPs act as an environmental stressor. Additionally, ML samples were centrifuged and the retention of MPs in the centrifuged solid (CS), which mainly depends on the size and morphology of MPs, was evaluated in relation to the amount and composition of EPSs. Remarkably, the concentration of humic acids showed a linear correlation with respect to the percentage of MPs retained in the CS. According to this, a possible mechanism based on hydrophobic interactions has been proposed as being responsible for the retention of MPs in sludge flocs.