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.

The contradictory roles of tightly bound and loosely bound extracellular polymeric substances of activated sludge in trimethoprim adsorption process.

Extracellular polymeric substances (EPS) of activated sludge are a mixture of high molecular weight polymers secreted by microorganisms, which have the double structure of tightly-bound EPS (TB-EPS) in inner layer and loosely-bound EPS (LB-EPS) in outer layer. The characteristic of LB- and TB-EPS were different, which would affect their adsorption of antibiotics. However, the adsorption process of antibiotics on LB- and TB-EPS was still unclear yet. Therefore, in this work, the roles of LB-EPS and TB-EPS in adsorption of a typical antibiotic-trimethoprim (TMP) at environmentally relevant concentration (25.0 µg/L) were investigated. The results showed the content of TB-EPS was higher than that of LB-EPS, which was 17.08 and 10.36 mg/g VSS, respectively. The adsorption capacity of raw, LB-EPS extracted and both LB- and TB-EPS extracted activated sludges for TMP were 5.31, 4.65 and 9.51 µg/g VSS, respectively, which indicated LB-EPS had positive effect on TMP removal, while TB-EPS had negative effect. The adsorption process can be well described by a pseudo-second-order kinetic model (R2 > 0.980). The ratio of different functional groups was calculated and the CO and C-O bond might be responsible for the adsorption capacity difference between LB- and TB-EPS. The fluorescence quenching results indicated that tryptophan protein-like substances in LB-EPS provided more binding sites (n = 0.36) than that of tryptophan amino acid in TB-EPS (n = 0.1). Furthermore, the extend DLVO results also demonstrated that LB-EPS promoted the adsorption of TMP, while TB-EPS inhibited the process. We hope the results of this study were helpful for understanding the fate of antibiotics in wastewater treatment systems.

Upgradation of sludge deep dewatering conditioners through persulfate activated by ferrous: Compatibility with sludge incineration, dewatering mechanism, ecological risks elimination and carbon emission performance.

Serious loss of organic substances and notable release of refractory intracellular organics and cell-free antibiotic resistance genes (ARGs) caused by cell lysis are found when quick lime, FeCl3, and cationic polyacrylamide (CPAM) were used as sludge conditioners, which is not feasible to sludge separate incineration and increases ecological risks. Therefore, persulfate oxidation through ferrous (Fe2+-Na2S2O8) activation was applied for the upgradation of sludge conditioner in China, the specific resistance to filtration (SRF) and capillary suction time (CST) significantly decreased and the removed water increased from 40% to 54%, implying that the persulfate activated by ferrous (PAF) conditioner presents good performance in sludge dewatering. Organic matter content and heating value of sludge merely decreased, and Cl- content in sludge simultaneously decreased with the use of the PAF conditioner, thereby effectively reducing the corrosion risk to the incinerator and showing good compatibility with sludge separate incineration. In accordance with ferrous activation, sulfate radical plays an important role in sludge dewatering process because remarkable decrease in polysaccharides and protein contents from tightly bound extracellular polymeric substances (TB-EPS) was discovered. Based on flow cytometry analysis, slight cell lysis presented better filtrate quality by the use of PAF conditioner, 49.3% of refractory intracellular organics was removed and the respective ermB, tetW and blaTEM decreased by factors of 37.3%, 54.5% and 63.6% due to the strong oxidizing property of sulfate radical. The intensive decrease in refractory intracellular organics and cell-free ARGs will reduce the ecological risks. The total carbon emission significantly decreases to 1771.1 kgCO2/tDS when PAF conditioner was employed, which is beneficial to the upgradation of sludge deep dewatering conditioners.

Flocculation with heterogeneous composition in water environments: A review.

Flocculation is a key process for controlling the fate and transport of suspended particulate matter (SPM) in water environments and has received considerable attention in the field of water science (e.g., oceanography, limnology, and hydrology), remaining an active area of research. The research on flocculation has been conducted to elucidate the SPM dynamics and to diagnose various environmental issues. The flocculation, sedimentation, and transportation of SPM are closely linked to the compositional and structural properties of flocs. In fact, flocs are highly heterogeneous in terms of composition. However, the lack of comprehensive research on floc composition and structure has led to misconceptions regarding the temporal and spatial dynamics of SPM. This review summarizes the current understanding of the heterogeneous composition of flocs (e.g., minerals, organic matter, metals, microplastic, engineered nanoparticles) and its effect on their structure and on their fate and transport within aquatic environments. Furthermore, the effects of human activities (e.g., pollutant discharge, construction) on floc composition are discussed.

[First Extended Anaerobic Phase Enhanced Nitrogen and Phosphorus Removal by Aerobic Granular Sludge Under Intermittent Gradient Aeration].

In this study, a SBR reactor was selected to explore the effect of nitrogen and phosphorus removal by aerobic granular sludge under different initial anaerobic time using intermittent gradient aeration (decreasing dissolved oxygen concentration in each aeration section) and actual domestic wastewater with low C/N ratio as the influent matrix. The results showed that the initial anaerobic stage of intermittent gradient aeration increased from 50 min to 90 min, which increased the carbon source reserves in granular sludge. This improved delayed anaerobic condition led to the improvement of nitrogen and phosphorus removal efficiency of granular sludge. When delayed to 70 and 90 min, the removal efficiency of COD, TN, and TP reached 84.74%, 70.05%, and 89.7% and 86.65%, 78.81%, and 85.5%, respectively. However, after the first anaerobic phase time was increased to 110 min, the sludge loss was more severe owing to the disintegration of some cells, leading to a decrease in internal carbon source reserves by about 13.6%. Owing to this, the pollutant removal efficiency decreased. In the process of prolonging the first anaerobic phase time from 50 min to 90 min, the content of PS in LB-EPS changed minimally; when it was delayed to 110 min, the content of PS increased to about 7.18 mg·g-1, and the PN content increased to about 5.56 mg·g-1. The contents of PN and PS in TB-EPS were stable, which indicated that the effect of internal carbon storage on LB-EPS was greater than that of TB-EPS, and the decrease in sludge settling performance was closely related to the increase in PS content in LB-EPS. The proportion of DPAOs in granular sludge increased as the first anaerobic phase time increased, and the proportion of DPAOs reached 51.5% when the first anaerobic phase time was 70 min.

Layered Extraction and Adsorption Performance of Extracellular Polymeric Substances from Activated Sludge in the Enhanced Biological Phosphorus Removal Process.

A large amount of phosphorus was found in the extracellular polymeric substances (EPS) of activated sludge used in enhanced biological phosphorus removal (EBPR), so the role of EPS and extracellular phosphorus in EBPR should not be neglected. The composition and properties of tightly bound EPS (TB-EPS) and loosely bound EPS (LB-EPS) were significantly different, and it was necessary to study the adsorption performance of EPS through the fractionating of activated sludge into LB-EPS, TB-EPS and microbial cells. In this study, the adsorption performance of LB-EPS and TB-EPS for phosphate was explored by extracting LB-EPS and TB-EPS via sonication and cation exchange resin (CER), respectively. The results indicated that the sonication-CER method was an efficient and reliable extraction method for EPS with a synergistic effect. The performance of EPS in the adsorption/complexing of phosphate was excellent because of its abundant functional groups. Specifically, the type and content of metal elements and functional groups in TB-EPS were much greater than those in LB-EPS, which led to the key role of TB-EPS in the adsorption/complexing of phosphate. Finally, a metabolic model for EBPR with consideration of the adsorption performance of LB-EPS and TB-EPS was proposed.

Deep dewatering process of sludge by chemical conditioning and its potential influence on wastewater treatment plants.

FeCl3, quick lime, and cationic polyacrylamide (CPAM) were used for excess sludge conditioning from wastewater treatment plant (WWTP) and the dewatering performance by different chemical conditioners was investigated. Experimental results showed that FeCl3 could make small and concentrated sludge particles. Furthermore, new mineral phase structures for building a dewatering framework were obtained by the addition of quick lime, and the coagulation capacity was enhanced by the formation of colloid hydroxyl polymer, which was induced due to the alkaline environment. In addition, the floc particle size significantly increased after the CPAM dosage. The bound water could be released with the stripping of tightly bound extracellular polymeric substance (EPS). Therefore, the dewatering performance and efficiencies were improved and subsequently the hypothetical sludge deep dewatering process was depicted in accordance with the variation of EPS. However, high-strength refractory organics in sludge filtrates caused by quick lime pyrolysis could lead to the unstable operation of the WWTP, because the relatively high concentrations of organic compounds with benzene were dominant in sludge dewatering filtrates.

Reactivation and pilot-scale application of long-term storage denitrification biofilm based on flow cytometry.

The work provides a method on the basis of flow cytometry to evaluate the performance of denitrification biofilm during the preservation, reactivation and pilot-scale operation process. The viable cell ratio of denitrification biofilm significantly reduced and further led to the decrease of denitrification capacity after long-term preservation for 5 months. Protein component in tightly bound extracellular polymeric substances (TB-EPS) could serve to enhance microbial adhesion and promote denitrification biofilm formation. With the significant correlation of viable cell ratio and microbial characteristics, 4 °C was more appropriate for preserving denitrification biofilm and conducive to maintain the relatively high denitrification capacity. A maximum denitrification rate of 5.80 gNO3--N/m2·d was obtained in pilot-scale anoxic-oxic (AO) process and Dechloromonas became greater prevalence in denitrification suspended carriers. Furthermore, the enrichment of Pseudomonas, Parcubacteria, Acidovorax, Aquabacterium and Unclassified_Flavobacteriaceae enhanced biofilm formation and nutrient conservation. The significantly positive correlation between viable cell ratio and the ratio of nitrate reduction to COD consumption was discovered, and the indices of Chao, ACE, Shannon and Simpson of denitrification biofilm were positively correlated with viable cell ratio, meaning that flow cytometry analysis was reasonable and suitable to evaluate the performances of denitrification biofilm.

The roles of loosely-bound and tightly-bound extracellular polymer substances in enhanced biological phosphorus removal.

Extracellular polymeric substances (EPS) have be founded to participate in the process of enhanced biological phosphorus removal (EBPR), but the exact role of EPS in EBPR process is unclear. In this work, the roles of loosely-bound EPS (LB-EPS), tightly-bound EPS (TB-EPS) and microbial cell in EBPR were explored, taking the activated sludge from 4 lab-scale A/O-SBR reactors with different temperatures and organic substrates as objects. It was founded that the P of EBPR activated sludge was mainly stored in TB-EPS, but the P of non-EBPR activated sludge was primarily located in microbial cell. The P release and uptake of EBPR activated sludge was attributed to the combined action of TB-EPS and microbial cell. Furthermore, TB-EPS played an more important role than microbial cell in EBPR process. With the analysis of 31P NMR spectroscopy, both polyP and orthoP were the main phosphorus species of TB-EPS in EBPR sludge, but only orthoP was the main phosphorus species of LB-EPS and microbial cell. During the anaerobic-aerobic cycle, the roles of LB-EPS, TB-EPS and microbial cell in transfer and transformation of P in EBPR sludge were obviously different. LB-EPS transported and retained orthoP, and microbial cell directly anaerobically released or aerobically absorbed orthoP. Importantly, TB-EPS not only transported and retained orthoP, but also participated in biological phosphorus accumulation. The EBPR performance of sludge was closely related with the polyp in TB-EPS, which might be synthesized and decomposed by extracellular enzyme.

Modeling optimization and evaluation of tightly bound extracellular polymeric substances extraction by sonication.

Tightly bound extracellular polymeric substances (TB-EPS) are important components of sludge, playing a crucial role in the behavior of activated sludge. They are located in the innermost layer of EPS which closely combine with the cell surface and are difficult to be extracted. To fully understand the role of TB-EPS, it is extremely important to find an appropriate TB-EPS extraction protocol, which provides maximum yields of TB-EPS under the premise of minimal contribution of cell lysis. Ultrasonic method has been widely applied for TB-EPS extraction due to its unique advantages, but no one has developed a systematic and scientifically optimized method for this protocol. In this study, a novel method based on response surface methodology (RSM) was successfully developed to optimize the conditions of TB-EPS extraction. The optimal conditions were determined at an ultrasound time of 1.00 min, ultrasonic density of 5.59 W/mL and a mixed liquor suspended solid (MLSS) of around 1700-1800 mg/L. Furthermore, combined analysis of microscopy, particle size, and excitation-emission matrix (EEM) was successfully applied to evaluate the optimal conditions. The result indicates that the optimal conditions are efficient, reliable, and reproducible which effectively solves the bottleneck problem of the conflict between cell viability and TB-EPS yield, and little effect on its chemical structure.