Tightly Bound EPS Research Articles

Enhancement of microalgae co-cultivation self-settling performance and water purificationcapacity of microalgae biofilm

Cultivating microalgae for the remediation of aquaculture wastewater provides a promising solution for pollution control. However, the economic viability of this approach faces challenges due to the high costs associated with microalgal biomass harvesting. This study aims to address this issue by immobilizing microalgae onto coral velvet carriers, enhancing the efficiency of biomass recovery. Four types of microalgae were screened: Chlorella sp., Isochrysis galbana, Chaetoceros sp., and Nannochloropsis sp. Among them, Isochrysis galbana exhibited the best self sedimentation rate, achieving a self sedimentation rate of 94.36%. Chlorella sp. demonstrated the best denitrification rate, with a nitrate removal rate of 100% and an inorganic nitrogen removal rate of 79.13%. In addition, this study found that extracellular polymeric substances(EPS) affects the self-settling performance of microalgae, and the results emphasize the key role of tightly-bound EPS(TB-EPS) content in determining self settling efficiency. Furthermore,the assessments of the purification of simulated aquaculture wastewater were conducted, comparing the outcomes of co-cultivation with mono-culture. The co-cultivation strategy showed exceptional efficacy, achieving a 100% removal rate for NO3−-N by the 5th day. In contrast, mono-cultures of Chlorella sp. and Isochrysis galbana showed removal rates of 77.76% and 45.72%, respectively, at the same interval. Applying of the co-cultivation microalgal biofilm to treat the actual aquaculture wastewater showed remarkable denitrification performance, attaining a 100% removal rate for NO3−-N by the 7th day. The study proposes the co-cultivation of Chlorella sp. and Isochrysis galbana for treating aquaculture wastewater and explores the potential application of immobilization technology to remove nitrogen-containing pollutants.

Different nitrogen conditions regulating extracellular polymeric substances and erosion resistance of sewer sediment: Mechanism of microbial metabolism and molecular response

Nitrogen biotransformation plays a vital role in the metabolism of microbial communities in sewers. Extracellular polymeric substances (EPS) secreted by microbial communities can form gel-like sewer sediments, causing clogging of the sewer. However, knowledge on the effects of varying nitrogen conditions on the erosion resistance of sewer sediments and EPS produced by sewer microorganisms is limited. In this study, two typical organic/inorganic nitrogen ratios of sewage were reproduced in simulated sewer reactors, i.e., 3/7 (R1 group) and 7/3 (R2 group). Higher organic nitrogen (ON) concentrations were found to increase the critical erosion shear stress by 26.43%; this was ascribed to increased particle diameter, weakened electrostatic repulsion of sediments and stimulated EPS secretion in the R2 group. The protein and polysaccharide contents of the R2 group were 48.84% and 34.25% higher than those of the R1 group, respectively, which was supported by increased gene abundances for aromatic amino acid synthesis, general secretory pathways of protein, and synthesis of precursors and polysaccharides. Tightly-bound EPS in R2 group exhibited increased contents of hydrophobic protein secondary structures and intermolecular hydrogen bonds, thereby promoting the formation of gel-like sediment structures with enhanced erosion resistance. However, the microbial diversity and the abundance of key genes involved in EPS generation and secretion (e.g., tyrB, yajC, secB, gumF, and gumH) obviously decreased in the R1 group. Moreover, high ON concentrations increased microbial diversity and enhanced microbial glycolysis, tricarboxylic acid cycle and ammonium assimilation. This study reveals the formation mechanisms of EPS in sewer sediments under different nitrogen conditions and their effects on sediment erosion resistance, which contributed to improved sewer system operation and sewer sediment control.

Exploring the regulation mechanism of signaling molecules on algal-bacterial granular sludge through different N-acyl-homoserine lactones

As a recently emerging wastewater treatment technology, Algal-bacterial granular sludge (ABGS) process shows significant advantages. However, current research on the ABGS system is a lack of a clear and complete understanding of the potential mechanism of signal molecules on the growth of ABGS. This study comprehensively explores the variations in the ABGS under different N-acyl-homoserine lactone (AHL)conditions by constructing three sequencing batch reactor (SBR) systems. The results indicate that N-hexanoyl-L-homoserine lactone (C6-HSL) accelerates the granulation process in the early stages by promoting the loosely bound extracellular polymeric substances (LB-EPS) secretion and filamentous bacteria growth, thereby shortening required time for initial granule formation. On the other hand, N-(3-oxodecanoyl)-L-homoserine lactone (3-oxo-C12-HSL) expedites the granulation process by promoting the tightly bound extracellular polymeric substances (TB-EPS) and aromatic protein secretion, benefiting structural stability and nitrogen and phosphorus removal efficiency of mature ABGS.

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.

Mixed microalgal culture enhances biofilm adhesion during treatment of livestock wastewater: The role of extracellular polymeric substances

Microalgal biofilm can remove nutrients and refractory pollutants from livestock wastewater, transforming them into biomass or fuel. However, these applications are hampered by biofilm shedding. Coculture enhances effects, and its influence on the adhesion of attached microalgal biofilms requires exploration. In this study, two unicellular algae, Euglena (E) and Chlorella pyrenoidosa (C). and two filamentous algae, Klebsodium sp. (K) and Phormidium sp. (P), were paired two by two, aiming to investigate the effect of mixing on the adhesion of microalgal biofilm. The results showed that suitable microalgal combinations significantly enhanced the performance of the co-cultured microalgal biofilm. Biomass enhancement of 35 % and 21.9 % was observed in the PC group compared to the individual P and C groups, respectively. The content of tightly bound extracellular polymeric substances (TB-EPS) and its protein and polysaccharide content significantly correlating with the net biomass increase in all treatments, p < 0.05. Furthermore, measurement of zeta potentials and water contact angles indicated that alterations in protein secondary structures contributed to a lower intermolecular repulsion and greater hydrophobicity of the TB-EPS, which together with the stability provided by the higher β-glucan content supported the adhesion of the co-cultured microalgal biofilm.

Understanding Partial Denitrification-hydroxyapatite (PD-HAP) coupled granules to enhance process stability and phosphorus removal by investigating the size effect of granules

The innovative Partial Denitrification-hydroxyapatite (PD-HAP) coupled granule offers a promising biotechnology for supplying nitrite to Anammox while simultaneously removing phosphorus. This study systematically examined the characteristics, biomass activity, and microbial community of PD-HAP granules of various sizes to enhance system stability and phosphorus removal efficiency. The results showed that larger granules had higher inorganic mineral and extracellular polymeric substances (EPS) content. These factors contributed to their denser structure, smoother surfaces, and better settling velocities compared to smaller granules. In contrast, smaller granules exhibited higher denitrification activity due to their greater biomass content and more efficient mass transfer. Calcium and phosphorus were identified as the primary inorganic constituents, mainly in the form of HAP, and were especially abundant in larger granules. Granules larger than 3.0 mm demonstrated a higher capacity for phosphorus removal. This was due to their significantly greater tightly bound EPS (T-EPS) content, which led to increased HAP and phosphorus levels. All granule sizes maintained a nitrate-to-nitrite transformation ratio (NTR) above 80 %, primarily due to the dominance of Flavobacterium (65.8 %-74.9 %). Although larger granules had lower denitrification activity, they still achieved an excellent NTR (above 85 %). They also showed superior settling velocity (174.9 m/h) and stronger phosphorus removal capacity (54.1 mg/g SS) compared to smaller granules. Maintaining a dominant proportion of granules larger than 3.0 mm could be an effective strategy to enhance both system stability and phosphorus removal. This study provides valuable insights into the PD-HAP coupled granular process, promoting efficient nitrite production and effective phosphorus removal.

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

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

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

Dissemination of antibiotic resistance genes (ARGs) has become a critical threat to public health. Activated sludge, rich in extracellular polymeric substances (EPS), is an important pool of ARGs. In this study, mechanisms of conjugation transfer of ARGs induced by EPS, including tightly bound EPS (TBEPS), soluble EPS (SEPS), and loosely bound EPS (LBEPS), were explored in terms of molecular diversities and electron transfer properties of EPS. Conjugation transfer frequency was increased by 9.98-folds (SEPS), 4.21-folds (LBEPS), and 15.75-folds (TBEPS) versus the control, respectively. Conjugation-related core genes involving SOS responses (9 genes), membrane permeability (18 genes), intercellular contact (17 genes), and energy metabolism pathways (13 genes) were all upregulated, especially in the presence of TBEPS. Carbohydrates and aliphatic substances in SEPS and LBEPS were contributors to ARG transfer, via influencing reactive oxygen species (ROS) formation (SEPS) and ROS and adenosine triphosphate (ATP) production (LBEPS). TBEPS had the highest redox potential and greatest lability and facilitated electron transfer and alternated respiration between cells, thus promoting ARG transfer by producing ATP. Generally, the chemical molecular characteristics and redox properties of EPS facilitated ARG transfer mainly by influencing lipid peroxidation and ATP, respectively.

Mechanisms of denitrifying granular sludge disintegration and calcium ion-enhanced re-granulation in acidic wastewater treatment

Granular sludge is an alternative technology for the direct treatment of acidic nitrate-containing wastewater. Rapid remediation of disintegrated granules is essential to achieve efficient nitrogen removal. In this study, denitrifying granules were inactivated and disintegrated when the influent nitrate-nitrogen concentration was elevated from 240 to 360 mg L−1 in acidic wastewater (pH = 4.1) in a sequencing batch reactor. Tightly bound extracellular polymeric substances (TB-EPS) decreased by 60%, and extracellular protein (PN) was the main component of the reduced EPS. The three-dimensional excitation emission matrices (3D-EEM) results confirmed that the PNs that decreased were mainly tryptophan-like, tyrosine-like, and aromatic. This study further confirmed that the decrease in PN was mainly from the destruction of C=O (amide I) and N–H functional groups. Overloading of nitrogen-inhibited denitrifying activity and the destruction and dissolution of TB-EPS by acidic pH were responsible for granule disintegration, with PNs playing a major role in maintaining granule stability. Based on this, new granules with an average particle size of 454.4 μm were formed after calcium chloride addition; EPS nearly doubled during granule formation with PN as the dominant component, accounting for 64.7–78.4% of the EPS. Atomic force microscopy (AFM) revealed that PN-PN adhesion increased by 1.6–4.9 times in the presence of calcium ions, accelerating the re-granulation of disintegrated particles. This study provides new insights into the disintegration and remediation of granular sludge under acidic conditions.

Comparative study of iron and aluminium coagulants in conditioning sludge: Sludge dewatering performance, physicochemical properties, and risk of heavy metal migration

Inorganic coagulants are essential agents that can facilitate the sludge deep dewatering, but comprehensive comparative research of commonly used coagulants is lacking. The systematic comparison of six coagulants, including Ferric Chloride (FeCl3), Polymeric Ferrous Sulfate (PFS), Ferrous Sulfate (Fe2(SO4)3), Aluminium Chloride (AlCl3), Aluminium Sulfate (Al2(SO4)3), and Polymeric Aluminium Chloride (PAC) regarding their performance in conditioning sludge dewatering, physicochemical properties, the risk of heavy metals migration in sludge cake and the mechanism of sludge dewatering, was investigated. Within a dosage range of 20% dry solids (DS), the PFS and FeCl3 showed a better dewatering effect among the six coagulants, and the water content (Wc) of the sludge cake decreased from 81.2%±1.1% to 57.7%±1.0% and 59.2%±1.6%, respectively. There was a correlation between the pH, zeta potential, viscosity, and bound water content of the sludge and the Wc of the sludge cake during dewatering conditioned by six coagulants. However, there was no correlation between sludge particle size and Wc of the sludge cake. The PFS and FeCl3 reduced the protein content of tightly bound extracellular polymeric substances (TB-EPS) by 66.1% and 62.6%, respectively. Moreover, the iron coagulants significantly reduced the content of As, Cu, Ni, and Zn in the sludge cake, where the risk levels of As and Ni were largely weakened. The economic analysis shows that iron coagulants were more valuable than aluminium coagulants for engineering applications. Meanwhile, the dewatering mechanisms of the two types of coagulants were revealed. This study provides significant insights into the application of coagulants in engineering practice.

New insights into algal-bacterial sludge granulation based on the tightly-bound extracellular polymeric substances regulation in response to N-Methylpyrrolidone

Algal-bacterial granular sludge (ABGS) system is promising in wastewater treatment for its potential in energy-neutrality and carbon-neutrality. However, traditional cultivation of ABGS poses significant challenges attributable to its long start-up period and high energy consumption. Extracellular polymeric substances (EPS), which could be stimulated as a self-defense strategy in cells under toxic contaminants stress, has been considered to contribute to the ABGS granulation process. In this study, photogranulation of ABGS by EPS regulation in response to varying loading rates of N-Methylpyrrolidone (NMP) was investigated for the first time. The results indicated the formation of ABGS with a maximum average diameter of ∼3.3 mm and an exceptionally low SVI5 value of 67 ± 2 mL g−1 under an NMP loading rate of 125 mg L−1 d−1, thereby demonstrating outstanding settleability. Besides, almost complete removal of 300 mg L−1 NMP could be achieved at hydraulic retention time of 48 h, accompanied by chemical oxygen demand (COD) and total nitrogen (TN) removal efficiencies higher than 90 % and 70 %, respectively. Moreover, possible degradation pathway and metabolism mechanism in the ABGS system for enhanced removal of NMP and nitrogen were proposed. In this ABGS system, the mycelium with network structure constituted by filamentous microorganisms was a prerequisite for photogranulation, instead of necessarily leading to granulation. Stress of 100–150 mg L−1 d−1 NMP loading rate stimulated tightly-bound EPS (TB-EPS) variation, resulting in rapid photogranulation. The crucial role of TB-EPS was revealed with the involved mechanisms being clarified. This study provides a novel insight into ABGS development based on the TB-EPS regulation by NMP, which is significant for achieving the manipulation of photogranules.

Quantitative image analysis for assessing extracellular polymeric substances in activated sludge under atrazine exposure

Extracellular polymeric substances (EPS) play a vital role in biological wastewater treatment systems. This study investigates the impact of herbicide atrazine (ATZ) on the overall performance, EPS yield, composition, and sludge morphology in an activated sludge (AS) system operated in a sequencing batch reactor (SBR). Since conventional methods for analyzing EPS are time-consuming and releases residues, a new approach was developed in this work to evaluate the EPS fractions and components, based on the morphological characterization of the biomass using quantitative image analysis (QIA) technique coupled with multivariate statistics.Results showed that exposure to ATZ inhibit biomass activity in terms of organic matter (COD) and nitrogen removal. Moreover, both tightly bound EPS (TB-EPS) and loosely bound EPS (LB-EPS) increased under ATZ, indicating that microorganisms release EPS as a defense mechanism against environmental changes. The PN/PS ratio also increases, indicating likely increased hydrophobicity in ATZ phases. Furthermore, ATZ phases exhibit a predominance of large aggregates compared to intermediate and small ones, indicating a change in aggregate morphological structure associated with EPS production. The new approach using QIA coupled with partial least squares (PLS) modeling provides accurate predictions of EPS content. The increase in TB-EPS is closely related to the rise of large aggregates in phases exposed to higher ATZ concentrations. The PLS models demonstrate high accuracy for EPS prediction (coefficients of determination, R2 above 0.86), showcasing the feasibility of using QIA for EPS assessment in AS systems. This approach offers significant potential for regular process monitoring and management, providing a more environmentally friendly methodology by eliminating the need for chemical usage.

Rapid conversion of heterotrophic denitrification to autotrophic denitrification in mariculture wastewater treatment: Denitrification performance and microbial communities under antibiotic stress

Mariculture wastewater, characterized by a low COD/Nitrogen (C/N) ratio and the occurrence of antibiotics, is commonly treated by a heterotrophic denitrification (HD) process with supplying substantial carbon sources. Compared to HD, sulfur-based autotrophic denitrification (AD) requires less external carbon sources and is more favorable for low-carbon wastewater treatment. However, the AD process often consumes more startup time. Thus, the present study was designed to explore a rapid startup strategy for the AD process by switching from HD to AD, treating mariculture wastewater. A rapid transformation from HD to AD was achieved by inoculating mixotrophic denitrifying sludge, reducing carbon sources and adding sulfur sources under the antibiotic sulfamethoxazole (SMX). From the functional microbial community point of view, our results showed that autotrophic bacteria exhibited greater adaptability to sulfamethoxazole than heterotrophic bacteria. During the transition from HD to AD, nitrate removal improved from 53.13 % to 68.57 %, and the secretion of tightly bound extracellular polymeric substances (TB-EPS) increased by 29.94 mg/g VSS. Meanwhile, in the heterotrophic phase, the presence of sulfate in mariculture wastewater maintained a high abundance of sulfate-reducing bacteria such as Desulfuromusa (0.8 %) and sulfur-based autotrophic denitrifying bacteria such as Sedimenticola (36.7 %) and Thiogranum (15.2 %). This increases the activity of sulfur metabolism during the heterotrophic phase and ensures a rapid transition from HD to AD. The rapid shift from HD to AD in seawater under antibiotic stress provides theoretical support for the improvement of mariculture wastewater treatment processes.

Deep dewatering of municipal sludge by combining alternating electric field electro-osmosis and nano-CaO2 oxidation

To improve the effect of traditional electroosmotic sludge dewatering, combining alternating electric field electro-osmosis and nano-calcium peroxide (CaO2) oxidation, a homemade vertical electro-osmosis dewatering device was employed in this work to study the effects of different parameter on the dewatering of primary dewatered sludge from municipal sewage treatment facilities with a moisture content of around 85%. The sludge water content (Wr) was used as an indicator, and concentration of extracellular polymeric substances (EPS) in the dewatered sludge was measured, and its structure was analyzed to investigate the effect of EPS changes on sludge dewatering.The response surface method was used to optimize the parameters. Wr decreased to 36.23 % at a CaO2 amount of 14.79 mg·g−1 dry sludge, a time ratio of 7.98:1, and a voltage gradient of 19.71 V·cm−1. Analysis of zeta potential and infrared characterization indicates that under acidic circumstances, CaO2 reacts more positively with water to produce hydrogen peroxide, and EPS is more easily destabilized and degraded, due to this, the problem of unequal water content in the cathode and anode sludge cakes is also solved. During the sludge dewatering process, the synergistic effect of alternating electric field and calcium peroxide together destroys the cell structure and EPS of sludge, especially the tightly bound EPS (TB-EPS), enabling the migration of substances and the formation of loosely bound EPS (LB-EPS), slime EPS (S-EPS), or filtrate, which can improve the sludge dewaterability.

Optimizing carbon sources regulation in the biochemical treatment systems for coal chemical wastewater: Aromatic compounds biodegradation and microbial response strategies

The complex composition of coal chemical wastewater (CCW), marked by numerous highly toxic aromatic compounds, induces the destabilization of the biochemical treatment system, leading to suboptimal treatment efficacy. In this study, a biochemical treatment system was established to efficiently degrade aromatic compounds by quantitatively regulating the dosage of co-metabolized substrates (specifically, the chemical oxygen demand (COD) Glucose: COD Sodium acetate = 3:1, 1:3, and 1:1). The findings demonstrated that the system achieved optimal performance under the condition that the ratio of COD Glucose to COD Sodium acetate was 3:1. When the co-metabolized substrate was added to the system at an optimal ratio, examination of pollutant removal and cumulative effects revealed that the removal efficiencies for COD and total organic carbon (TOC) reached 94.61 % and 86.40 %, respectively. The removal rates of benzene series, nitrogen heterocyclic compounds, polycyclic aromatic hydrocarbons, and phenols were 100 %, 100 %, 63.58 %, and 94.12 %, respectively. Research on the physiological response of microbial cells showed that, under optimal ratio regulation, co-metabolic substrates led to a substantial rise in microbial extracellular polymeric substances (EPS) secretion, particularly extracellular proteins. When the system reached the end of its operation, the contents of loosely bound EPS (LB-EPS) and tightly bound EPS (TB-EPS) for proteins in the optimal group were 7.12 mg/g-SS and 152.28 mg/g-SS, respectively. Meanwhile, the ratio of α-Helix / (β-Sheet + Random coil) and the proportion of intermolecular interaction forces were also increased in the optimal group. At system completion, the ratio of α-Helix / (β-Sheet + Random coil) reached 0.717 (LB-EPS) and 0.618 (TB-EPS), respectively. Additionally, the proportion of intermolecular interaction forces reached 74.83 % (LB-EPS) and 55.03 % (TB-EPS). An in-depth analysis of the metabolic regulation of microorganisms indicated that the introduction of optimal ratios of co-metabolic substrates contributed to a noteworthy upregulation in the expression of Catechol 2,3-dioxygenase (C23O) and Dehydrogenase (DHA). The expression levels of C23O and DHA were measured at 0.029 U/mg Pro·g MLSS and 75.25 mg TF·(g MLSS·h)−1 (peak value), respectively. Correspondingly, enrichment of aromatic compound-degrading bacteria, including Thauera, Saccharimonadales, and Candidatus_Competibacter, occurred, along with the upregulation of associated functional genes such as Catechol 1,2-dioxygenase, Catechol 2,3-dioxygenase, Protocatechuate 3,4-dioxygenase, and Protocatechuate 4,5-dioxygenase. Considering the intricate system of multiple coexisting aromatic compounds in real CCW, this study not only obtained an optimal ratio for carbon source addition but also enhanced the efficient utilization of carbon sources and improved the capability of the system to effectively degrade aromatic compounds. Additionally, this paper established a theoretical foundation for metabolic regulation and harmless treatment within the biochemical treatment of intricate systems, exemplified by real CCW.

Metabolomic reveals the responses of sludge properties and microbial communities to high nitrite stress in denitrifying phosphorus removal systems

Nitrite, as an electron acceptor, plays a good role in denitrifying phosphorus removal (DPR); however, high nitrite concentration has adverse affects on sludge performance. We investigated the precise mechanisms of responses of sludge to high nitrite stress, including surface characteristics, intracellular and extracellular components, microbial and metabolic responses. When the nitrite stress reached 90 mg/L, the sludge settling performance was improved, but the activated sludge was aging. FTIR and XPS analysis revealed a significant increase in the hydrophobicity of the sludge, resulting in improve settling performance. However, the intracellular carbon sources synthesis was inhibited. In addition, the components in the tightly bound extracellular polymeric substances (TB-EPS) of sludge were significantly reduced and indicated the disturb of metabolism. Notably, Exiguobacterium emerged as a new genus when face high nitrite stress that could maintaining survival in hostile environments. Moreover, metabolomic analysis demonstrated strong biological response to nitrite stress further supported above results that include the inhibited of carbohydrate and amino acid metabolism. More importantly, some lipids (PS, PA, LysoPA, LysoPC and LysoPE) were significantly upregulated that related enhanced membrane lipid remodeling. The comprehensive analyses provide novel insights into the high nitrite stress responses mechanisms in activated sludge systems.

Evaluating the role of carbon sources on the development of algal-bacterial granular sludge: From sludge characteristics, extracellular polymer properties, quorum sensing, and microbial communities

This study investigated the effect of dissolved organic matter (DOM) and particulate organic matter (POM) on the formation process of algal-bacterial granular sludge (ABGS). The results showed that compared with DOM, POM accelerated biomass growth and improved the settling performance of the sludge. However, concerning pollutant removal, ABGS cultured with DOM exhibited high TN and TP removal efficiency. The analysis of extracellular polymeric substances (EPS), indicated that the presence of POM primarily facilitated the production of loosely bound EPS (LB-EPS), whereas the presence of DOM promoted the secretion of tightly bound EPS (TB-EPS). Notably, carbon sources influenced the secretion of different signal molecules. ABGS cultured with POM exhibited higher concentrations of 3-oxo-C8-HSL than that cultivated with DOM, while DOM resulted in the production of more C8-HSL and C6-HSL. Microbial communities analysis revealed that the dominant bacteria in the mature ABGS cultured with different carbon sources were Plasticicumulans, while the predominant algal genus displayed notable diversity. Further investigations into functional genes suggested that the nitrification process (hao) and the denitrification process (nir, nar) were greatly enhanced in the presence of DOM. This study reveals the mechanism of carbon source types and ABGS formation, providing valuable guidance for the treatment of municipal wastewater with high POM proportion.

Concurrent removal of carbon and nutrients in a one-stage dual internal circulation airlift A2O bioreactor from milk processing industrial wastewater: Process optimization, sludge characteristics and operating cost evaluation

In this work, a one-stage dual internal circulation airlift anaerobic/anoxic/aerobic (DCAL-A2O) bioreactor was continuously operated for concurrent removal of nutrients and organics from milk processing wastewater (MPW). Special configuration of the airlift A2O bioreactor created possibility of the formation of desired anaerobic, anoxic and aerobic zones in a single unit. The process functionality of the bioreactor was examined under three influential operating variables i.e. hydraulic retention time (HRT; 7–15 h), air flow rate (AFR; 1–3 L/min) and aerobic volume ratio (AVR; 0.324–0.464). The optimum region was identified at HRT of 13h, AFR of 2L/min and AVR of 0.437, leading to TCOD, TN and TP removal efficiency of 94.5 %, 59.6 %, and 62.2 %, respectively, and effluent turbidity of 8 NTU. The impact of feed biodegradability on the process performance of the bioreactor treating the MPW, soft drink wastewater (SDW) and soybean oil plant wastewater (SOW) was also assessed. From the results, the feed characteristics affected significantly the nutrients removal. Moreover, the feeding location played an effective role in the nutrient removal while treating the MPW at optimum operating conditions. In this study, the change in residual organic matters as soluble microbial products (SMP) was monitored at various operating conditions. In addition, the impact of SMP extracted from sludge, extracellular polymeric substances (EPS) comprising of loosely bound EPS (LB-EPS) and tightly bound EPS (TB-EPS) was analyzed on sludge characteristics as bio-flocculation and settleability properties. According to the obtained data, the increase in operating variables led to the reduction in contents of effluent SMP, sludge SMP, LB-EPS, turbidity, and SVI, thereby, the enhancement in the sludge characteristics. Meanwhile, analysis of microbial communities verified the presence of various functional bacterial species. The cost operating evaluation confirmed the cost effectiveness of the airlift A2O bioreactor in reduction of energy consumption for the MPW treatment.

Effect of extracellular polymeric substances on the colony size and morphological changes of Microcystis.

Surface blooms of colony-forming Microcystis are increasingly occurring in aquatic ecosystems on a global scale. Recent studies have found that the Microcystis colonial morphology is a crucial factor in the occurrence, persistence, and dominance of Microcystis blooms, yet the mechanism driving its morphological dynamics has remained unknown. This study conducted a laboratory experiment to test the effect of extracellular polymeric substances on the morphological dynamics of Microcystis. Ultrasound was used to disaggregate colonies, isolating the cells and of the Microcystis suspension. The single cells were then re-cultured under three homologous EPS concentrations: group CK, group Low, and group High. The size, morphology, and EPS [including tightly bound EPS (TB-EPS), loosely bound EPS (LB-EPS), bound polysaccharides (B-polysaccharides), and bound proteins (B-proteins)] changes of colonies were closely monitored over a period of 2 months. It was observed that colonies were rapidly formed in group CK, with median colony size (D50) reaching 183 µm on day 12. The proportion of colonies with a size of 150-500 µm increased from 1% to more than 50%. Colony formation was also observed in both groups Low and High, but their D50 increased at a slower rate and remained around 130 µm after day 17. Colonies with a size of 50-150 µm account for more than 50%. Groups CK and Low successively recovered the initial Microcystis morphology, which is a ring structure formed of several small colonies with a D50 of 130 µm. During the recovery of the colony morphology, the EPS per cell increased and then decreased, with TB-EPS and B-polysaccharides constituting the primary components. The results suggest that colony formation transitioned from adhesion driven to being division driven over time. It is suggested that the homologous EPS released into the ambient environment due to the disaggregation of the colony is a chemical cue that can affect the formation of a colony. This plays an important but largely ignored role in the dynamics of Microcystis and surface blooms.

Effect of electrolytic zero-valent iron activated sodium hypochlorite on sludge dewatering performance.

Using electrolytic zero-valent iron-activated sodium hypochlorite (EZVI-NaClO) to pretreat sludge, the capillary suction time (CST) was utilized to evaluate sludge dewaterability. Ammonia nitrogen (NH4-N), dissolved phosphorus, and total phosphorus in the supernatant were used to analyze sludge disintegration. This approach aimed to evaluate the effectiveness of the pretreatment process and its impact on the sludge composition. The migration and transformation of extracellular polymeric substances (EPS), including dissolved EPS (S-EPS), loosely boundEPS, and tightly bound-EPS (TB-EPS), were analyzed by detecting protein and polysaccharide concentrations and three-dimensional fluorescence excitation-emission spectroscopy (3D-EEM). The sludge particle properties, including sludge viscosity and particle size, were also analyzed. The results suggested that the optimal pH value, NaClO dosage, current, and reaction time were 2, 100 mg/gDS (dry sludge), 0.2A, and 30 min, respectively, with a CST reduction of 43%. Protein and polysaccharide contents in TB-EPS were significantly reduced in the EZVI-NaClO group. Conversely, protein and polysaccharides contents in S-EPS increased, suggesting that EZVI-NaClO treatment could disrupt the EPS. Besides, the viscosity of the treated sludge decreased from 195.4 to 54.9 mPa·S, indicating that sludge fluidity became better. ZEVI-NaClO could enhance sludge dewaterability by destructing protein and polysaccharide structure and improving sludge hydrophobicity.