Influence of loosely bound extracellular polymeric substances (EPS) on the flocculation, sedimentation and dewaterability of activated sludge

Laboratory experiments on the activated sludge (AS) process were carried out to investigate the influence of microbial extracellular polymeric substances (EPS), including loosely bound EPS (LB-EPS) and tightly bound EPS (TB-EPS), on biomass flocculation, sludge settlement and dewaterability. The heat EPS extraction method was modified to include a mild step and a harsh step for extracting the LB-EPS and TB-EPS, respectively, from the sludge suspension. Six lab-scale AS reactors were used to grow AS with different carbon sources of glucose and sodium acetate, and different sludge retention times (SRTs) of 5, 10 and 20 days. The variation in the bioreactor condition produced sludge with different abundances of EPS and different flocculation and separation characteristics. The sludge that was fed on glucose had more EPS than the sludge that was fed on acetate. For any of the feeding substrates, the sludge had a nearly consistent TB-EPS value regardless of the SRT, and an LB-EPS content that decreased with the SRT. The acetate-fed sludge performed better than the glucose-fed sludge in terms of bioflocculation, sludge sedimentation and compression, and sludge dewaterability. The sludge flocculation and separation improved considerably as the SRT lengthened. The results demonstrate that the LB-EPS had a negative effect on bioflocculation and sludge-water separation. The parameters for the performance of sludge-water separation were much more closely correlated with the amount of LB-EPS than with the amount of TB-EPS. It is argued that although EPS is essential to sludge floc formation, excessive EPS in the form of LB-EPS could weaken cell attachment and the floc structure, resulting in poor bioflocculation, greater cell erosion and retarded sludge-water separation.     

Relationship between floc composition and flocculation and settling properties studied at a full scale activated sludge plant

The variation in activated sludge floc composition, flocculation and settling properties was studied at a full scale plant over a 2-year period. A comprehensive set of process parameters was analysed and related to the floc properties to increase the understanding of the factors affecting floc formation. The composition of the activated sludge showed a seasonal change with higher concentrations of extractable extracellular polymeric substances (EPS) during the winter months. The protein content of the total sludge and EPS increased significantly during the winter. This coincided with higher effluent suspended solids concentrations and increased shear sensitivity of the sludge flocs. Only poor correlations between EPS contents and stirred sludge volume index (SSVI) could be observed. High iron concentrations in the sludge due to dosage of iron salt to precipitate phosphorus were found to have a negative impact on the settling and compaction properties of the sludge, whereas it had a positive impact on floc stability. Higher organic loading due to by-passed primary settlers leads to improved settling and compaction properties.     

Metal removal in activated sludge: the role of bacterial extracellular polymers

A combination of flocculation and settling is the mechanism by which metal removal is achieved in activated sludge. Bacterial extracellular polymers appear to play an important role in flocculation; metal cations may also be involved in this process. Extracellular polymers in activated sludge are mainly of a polysaccharide nature, although protein and nucleic acid from autolysis are constituents of the polymer matrix. Precipitated metals may be removed either by independent settling or by physical trapping in the sludge floc matrix. Metals present in the ionic form may be removed from solution by adsorption to sites on bacterial extracellular polymers. Metal ions may also be accumulated in the cytoplasm of a bacterial cell, or adsorbed on to the cell wall.If activated sludge plants are overloaded with metals, toxic effects on bacteria and other microorganisms may severely inhibit the treatment process, resulting in poor quality effluents. Acclimated bacterial cultures can tolerate much higher metal concentrations than non-acclimated cultures; these are advantageous in the treatment of metal-laden wastes.     

Gravitational sedimentation of flocculated waste activated sludge

The sedimentation characteristics of flocculated wastewater sludge have not been satisfactorily explored using the non-destructive techniques, partially owing to the rather low solid content (ca. 1-2%) commonly noted in the biological sediments. This paper investigated, for the first time, the spatial-temporal gravitational settling characteristics of original and polyelectrolyte flocculated waste activated sludge using Computerized Axial Tomography Scanner. The waste activated sludge possessed a distinct settling characteristic from the kaolin slurries. The waste activated sludges settled more slowly and reached a lower solid fraction in the final sediment than the latter. Flocculation markedly enhanced the settleability of both sludges. Although the maximum achievable solid contents for the kaolin slurries were reduced, flocculation had little effects on the activated sludge. The purely plastic rheological model by Buscall and White (J Chem Soc Faraday Trans 1(83) (1987) 873) interpreted the consolidating sediment data, while the purely elastic model by Tiller and Leu (J. Chin. Inst. Chem. Eng. 11 (1980) 61) described the final equilibrated sediment. Flocculation produced lower yield stress during transient settling, thereby resulting in the more easily consolidated sludge than the original sample. Meanwhile, the flocculated activated sludge was stiffer in the final sediment than in the original sample. The data reported herein are valuable to the theories development for clarifier design and operation.     

Correlation of EPS content in activated sludge at different sludge retention times with membrane fouling phenomena

In this study, activated sludge characteristics were studied with regard to membrane fouling in membrane bioreactors (MBRs) for two pilot plants and one full-scale plant treating municipal wastewater. For the full-scale MBR, concentrations of extracellular polymeric substances (EPS) bound to sludge flocs were shown to have seasonal variations from as low as 17mgg(-1) dry matter (DM) in summer up to 51mg(gDM)(-1) in winter, which correlated with an increased occurrence of filamentous bacteria in the colder season. Therefore, it was investigated at pilot-scale MBRs with different sludge retention times (SRTs) whether different EPS contents and corresponding sludge properties influence membrane fouling. Activated sludge from the pilot MBR with low SRT (23d) was found to have worse filterability, settleability and dewaterability. Photometric analysis of EPS extracts as well as LC-OCD measurements showed that it contained significantly higher concentrations of floc-bound EPS than sludge at higher SRT (40d) The formation of fouling layers on the membranes, characterised by SEM-EDX as well as photometric analysis of EPS extracts, was more distinct at lower SRT where concentrations of deposited EPS were 40-fold higher for proteins and 5-fold higher for carbohydrates compared with the membrane at higher SRT. Floc-bound EPS and metals were suggested to play a role in the fouling process at the full-scale MBR and this was confirmed by the pilot-scale study. However, despite the different sludge properties, the permeability of membranes was found to be similar.     

A combined approach for a better understanding of wastewater treatment plants operation: statistical analysis of monitoring database and sludge physico-chemical characterization

Biological wastewater treatment plants (WWTP) are complex systems to assess. Many parameters are recorded daily in WWTP to monitor and control the treatment process, providing huge amounts of registered data. A combined approach of extracting information from the WWTP databases by statistical methods and from the sludge physico-chemical characterization was used here for a better understanding of the WWTP operation. The monitored parameters were analysed by multivariate statistical methods: Principal Components Analysis and multiple partial linear regression. The WWTP operational conditions determine the sludge characteristics. The bacterial activity of the sludge in terms of extra-cellular polymeric substances (EPS) production was assessed using size exclusion chromatography and the internal structure of sludge flocs was observed by confocal laser scanning microscopy. The diagnosis of three paper mill WWTP enabled the identification of an important EPS production, the presence of the nitrification process and the presence of PO₄³⁻ nutrient in WWTP-A. These three main characteristics of WWTP-A were related with a systematically good sludge settling. In WWTP-B and C with bad settling, the bacterial activity was weak.     

The influence of key chemical constituents in activated sludge on surface and flocculating properties

This paper examines the influence of the chemical constituents of activated sludge and extracted extracellular polymeric substances (EPS) on the surface properties, hydrophobicity, surface charge (SC) and flocculating ability (FA) of activated sludge flocs. Activated sludge samples from 7 different full-scale wastewater treatment plants were examined. Protein and humic substances were found to be the dominant polymeric compounds in the activated sludges and the extracted EPS, and they significantly affected the FA and surface properties, hydrophobicity and SC, of the sludge flocs. The polymeric compounds proteins, humic substances and carbohydrates in the sludge flocs and the extracted EPS contributed to the negative SC, but correlated negatively to the hydrophobicity of sludge flocs. The quantity of protein and carbohydrate within the sludge and the extracted EPS was correlated positively to the FA of the sludge flocs, while increased amounts of humic substances resulted in lower FA. In contrast, increased amounts of total extracted EPS had a negative correlation to FA. The results reveal that the quality and quantity of the polymeric compounds within the sludge flocs is more informative, with respect to understanding the mechanisms involved in flocculation, than if only the extracted EPS are considered. This is an important finding as it indicates that extracting EPS may be insufficient to characterise the EPS. This is due to the low extraction efficiency and difficulties involved in the separation of EPS from other organic compounds. Correlations were observed between the surface properties and FA of the sludge flocs. This confirms that the surface properties of the sludge flocs play an important role in the bioflocculation process but that also other interactions like polymer entanglement are important.     

Characterization of activated sludge exopolymers from various origins: a combined size-exclusion chromatography and infrared microscopy study

High-pressure size-exclusion liquid chromatography and infrared microscopy were coupled to investigate the molecular weight and nature of extracellular polymeric substances (EPS) from various activated sludges. Six main families of compounds (proteins, polysaccharides, organic acids, lipids, mineral phases) were found either as a single molecule or as associations. The molecular weight of proteins varied from small (10 kDa) to large (600 kDa) sizes, while all polysaccharides were smaller than 1 kDa. Association of different molecules implied the presence of species large in size. The EPS chromatographic fingerprints of sludge from various origins remained stable in normal operating conditions, but were drastically modified during settling crises. In poor settling conditions, the EPS with smaller molecular sizes always prevailed and large polymers were underrepresented. The EPS identified in activated sludge were collected in a chemical database which provides the basis for comparison of municipal and industrial wastewater treatment plants (WWTP).     

Extracellular proteins, polysaccharides and enzymes impact on sludge aerobic digestion after ultrasonic pretreatment

Ultrasonic pretreatment of excess sludge can improve its aerobic digestibility, leading to enhanced sludge reduction. In order to understand the mechanisms of this improvement, sludge flocs were divided into four layers, i.e. (1) slime, (2) loosely bound extracellular polymeric substances (LB-EPS), (3) tightly bound EPS (TB-EPS) and (4) pellet. Extracellular proteins, polysaccharides and five types of hydrolytic enzymes (protease, alpha-amylase, alpha-glucosidase, alkaline-phosphatase and acid-phosphatase) from sludge flocs were investigated to determine their influence on sludge aerobic digestion after ultrasonic pretreatment. Results suggested that most of the extracellular enzymes (except alpha-amylase) were present in pellet and TB-EPS layers, with minor quantities detected in LB-EPS and slime layers, and almost none detected in bulk solution. As for alpha-amylase in sludge flocs, most of it (52.6%) was also mainly bound with pellet; however, the rest of it was dispersed nearly uniformly throughout the sludge flocs. Ultrasonic pretreatment enhances enzymatic activities and promotes the shifts of extracellular proteins, polysaccharides and enzymes from inner layers of sludge flocs, i.e., pellet and TB-EPS, to outer layers, i.e., slime, to increase the contact and interaction among extracellular proteins, polysaccharides and enzymes that were originally embedded in the sludge flocs, resulting in improved efficiency in aerobic digestion. The optimum ultrasonic pretreatment conditions had a lasting time of 10min and density of 3 kWL(-1) at the frequency of 20 kHz. With the optimum ultrasonic pretreatment, the sludge reduction for TSS in aerobic digestion was 42.7% in which the part of 11.8% was removed by the ultrasonic pretreatment, compared with 20.9% for control, after an aerobic digestion time of 10.5d.     

Sludge settling and algal flocculating activity of extracellular polymeric substance (EPS) derived from bacillus cereus SK

Exopolysaccharides (EPS) of microbial origin is a promising alternative to chemical flocculants due to their sustainability and eco‐friendly nature. In the present study, EPS produced by Bacillus cereus SK was studied and their flocculating activity was investigated. On incubation for 48 h, B. cereus SK produced 0.919 ± 0.022 g/L of EPS from a cell mass of 3.59 ± 0.06 g/L. The isolated EPS was identified as glycoprotein and chemical characterization by FTIR spectroscopic analysis suggested the presence of hydroxyl, amino and carboxyl groups. The kaolin clay flocculating activity of 83.4% was obtained on treatment with 12 mg/L of EPS. At 30% (v/v) of EPS 31.22 ± 4.71 mL/g of Sludge volume index (SVI) and 34.44 ± 1.51 mL/g of algal flocculation was observed. The EPS produced by B. cereus SK was non‐toxic and did not exert any adverse effect on Danio rerio even at a concentration of 800 mg/mL.     

Novel insights into sludge dewaterability by fluorescence excitation-emission matrix combined with parallel factor analysis

Sludge dewatering is of major interest in sludge volume reduction and handling properties improvement. Here we report an approach of fluorescence excitation-emission matrix (EEM) combined with parallel factor (PARAFAC) analysis to elucidate the factors that influence sludge dewaterability. Sludge flocs from 11 full-scale wastewater treatment plants were collected to stratify into different extracellular polymeric substances (EPS) fractions and then to characterize their fluorescence EEMs. Both the normalized capillary suction time (CST) and specific resistance to filtration (SRF) were applied to determine sludge dewaterability. The results showed that fluorescence EEMs of tightly bound fractions were not affected by the wastewater sources. In contrast, fluorescence EEMs of loosely bound fractions were affected by the wastewater sources. All the fluorescence EEMs could be successfully decomposed into a six-component model by PARAFAC analysis. Both normalized CST and SRF were significantly correlated with component 1 [excitation/emission (Ex/Em) = (220, 275)/350] in the supernatant fraction, with components 5 [Ex/Em = (230, 280)/430] and 6 [Ex/Em = (250, 360)/460] in the slime and LB-EPS fraction. These results reveal that except for proteins-like substances (component 1), sludge dewaterability is also affected by humic acid-like and fulvic acid-like substances (components 5 and 6) in the slime and LB-EPS fractions. Furthermore, this paper presents a promising and facile approach (i.e., EEM-PARAFAC) for investigating sludge dewaterability.     

Lysis as a factor in sludge flocculation

A tentative relationship was established between the levels of extractable exopolymers and flocculation in activated sludge growing on a carbon limited system.

Deflocculation occurred in the logarithmic growth phase and flocculation predominated in the endogenous phase. There appeared to be a relationship between lysis of sludge and flocculation, as a decrease in cell dry weight accompanied by the release of polymers, e.g. nucleic acids, resulted in an increase in flocculation.

This relationship was further investigated using selected components released on lysis, i.e. DNA, which was shown to promote significant levels of flocculation when added to various microbial systems—a significant increase in the rate of flocculation in the case of addition to sludge and in increase in the rate and degree of flocculation when added to individual bacterial cultures. Thus DNA can act as a floc agent and can also survive breakdown or depolymerization. Thus polymers released on lysis of cells can enhance flocculation.     

Effects of temperature and dissolved oxygen on sludge properties and their role in bioflocculation and settling

Effects of temperature (mesophilic (35 °C) vs. thermophilic (55 °C)) and dissolved oxygen (DO) concentration (under thermophilic conditions) on sludge properties and their role in bioflocculation and settling were studied using well-controlled sequencing batch reactors fed with a synthetic wastewater comprised of glucose and inorganic nutrients. Under a similar DO level, thermophilic sludge had a poorer bioflocculating ability and settleability than that of mesophilic sludge. Under a thermophilic condition, an increase in DO level led to a poorer settleability and a slightly improved bioflocculating ability. A poorer settleability was related to a higher level of filaments. Analysis of bound extracellular polymeric substances (EPS) indicates that thermophilic sludge had a higher level of total bound EPS content than that of mesophilic sludge under a similar DO level, and an increase in DO resulted in an increase in total bound EPS content in thermophilic sludge. Surface analysis of sludge by X-ray photoelectron spectroscopy (XPS) suggests that significant differences in the surface concentrations of elements N, C, O were observed between thermophilic and mesophilic sludge, implying significant differences in bound EPS composition. The results of gel permeation chromatography indicate that the weight-averaged molecular weight (M_w) of bound EPS covered a range of 1159 Da to 13220 Da. The distribution of EPS “species” at floc surfaces was shown by transmission electron microscopy (TEM) to be uneven; different kinds of nanoscale materials were distributed in a patchy manner at the floc-water interface. The results suggest that it is the role of specific EPS molecules rather than the quantity of bound EPS that determine the difference in bioflocculation behavior between thermophilic and mesophilic sludge. The strategy of increasing the DO level could not solve the biomass separation problems associated with thermophilic sludge.     

Exopolymeric production by bacterial strains isolated from activated sludge of paper industry

The bacteria originated from the paper mill sludge were identified, isolated and cultured. Enterobacter (E) and Klebsiella (K) represented 70% of the culturable aerobic flora. Kinetics of EPS production in isolated culture and in arbitrary 1:1 mixture was investigated by high-pressure size exclusion chromatography (HPSEC). In isolated culture, Enterobacter produced regularly and gradually during 24 h of culture, while Klebsiella produced the total amount of EPS already after 4 h without any changes later. The chromatographic profiles of the EPS production of the 1:1 mixture after 4, 8 and 24 h of culture could be fitted as a linear combination of the profiles of exopolymers produced by pure Enterobacter and Klebsiella strains. After 24 h of culture, Enterobacter was predominant in term of population (82%) and produced about 77% EPS amount, in spite of competition phenomena observed during culture.     

Roles of extracellular polymeric substances (EPS) in the migration and removal of sulfamethazine in activated sludge system

The occurrences, transformation of antibiotics in biological wastewater treatment plants have attracted increasing interests. However, roles of extracellular polymeric substances (EPS) of activated sludge on the fate of antibiotics are not clear. In this study, the roles of EPS in the migration and removal of one typical antibiotic, sulfamethazine (SMZ), in activated sludge process were investigated. The interaction between EPS and SMZ was explored through a combined use of fluorescence spectral analysis, laser light scattering and microcalorimetry techniques. Results show that SMZ interacted with the proteins in EPS mainly with a binding constant of 1.91 × 10⁵ L/mol. The binding process proceeded spontaneously, and the driving force was mainly from the hydrophobic interaction. After binding, the structure of EPS was expanded and became loose, which favored the mass transfer and pollution capture. The removal of SMZ was influenced by interaction with EPS. SMZ could be effectively adsorbed on EPS, which accounted for up to 61.8% of total SMZ adsorbed by sludge at the initial adsorption stage and declined to around 35.3% at the subsequent biodegradation stage. The enrichment of SMZ by EPS was beneficial for SMZ removal and acquisition by microbes at the subsequent biodegradation stage.     

Morphological characterisation of ATAD thermophilic sludge; sludge ecology and settleability

Autothermal thermophilic aerobic digestion (ATAD) is a biological wastewater treatment process used for stabilisation of domestic, animal, food and pharmaceutical sludges, and wastewater. It produces a high-quality effluent due to thermophilic processing conditions, however the stabilised sludge has poor settling characteristics, a high water content, low compaction capacity and is difficult to dewater by mechanical processes alone. These factors impact transport and disposal of processed ATAD sludge. We have carried out a detailed morphological characterisation of ATAD sludge at all stages of the ATAD process in an attempt to determine key characteristics of the sludge that might be responsible for its poor dewatering and settleability. A number of microscopic techniques including electron, optical, wide field and laser scanning confocal microscopy were applied to fresh, fixed or embedded sludge taken at various stages during a full scale ATAD process treating domestic sludge. The spatial distributions of structural sludge matrix components were determined and suggested a highly dynamic sludge morphology during the overall process. Large amounts of fibres were observed in the feed sludge, whereas thermophilic sludge liquor with low settleability was shown to have a lower protein to polysaccharide ratio (1:0.9) compared to the easily settled fraction where ratio values were in the range of (1:1.14-1:1.7) with a prevalence of protein constituents. ATAD sludge was also shown to contain colloids, slime, cellulose micro-particles and multiple hydrophobic droplets in the bulk liquor, factors that may markedly impact on sludge dewaterability characteristics. Laser scanning confocal microscopy demonstrated a superior ability to identify composition and spatial localisation of structural constituents in such a dispersed, high water content sludge.     

改性聚苯板与传统聚苯板的性能特点分析

解释了膨胀聚苯乙烯泡沫板(EPS板)、石墨模塑聚苯乙烯泡沫塑料板(SEPS板)和热固性改性聚苯乙烯泡沫塑料板(热固性EPS板)的定义,对6个不同厂家生产的EPS板、SEPS板、热固性EPS板的主要物理性能进行了研究,得出了一些有实用价值的结论。     

Filtration properties of activated sludge in municipal MBR wastewater treatment plants are related to microbial community structure

In the conventional activated sludge process, a number of important parameters determining the efficiency of settling and dewatering are often linked to specific groups of bacteria in the sludge – namely floc size, residual turbidity, shear sensitivity and composition of extracellular polymeric substances (EPS). In membrane bioreactors (MBRs) the nature of solids separation at the membrane has much in common with sludge dewaterability but less is known about the effect of specific microbial groups on the sludge characteristics that affect this process.     

Enhanced sludge dewatering by dual polyelectrolytes conditioning

Sludge dewatering by dual polyelectrolytes conditioning was investigated in this study. Single polyelectrolyte is utilized in sludge conditioning conventionally, in which charge neutralization and bridging are involved in the reactions. In the current study, both cationic and non-ionic polyelectrolytes were utilized simultaneously in the conditioning. Waste activated sludge was sampled from a synthetic fiber plant, and used in the experiment. Capillary suction time (CST), specific resistance to filtration (SRF), and settling rate were used to evaluate the sludge dewatering behaviors. Zeta potentials were also measured. Sludge dewatering behaviors resulting from single and dual polyelectrolytes conditioning were compared. It was found that sludge conditioned with dual polyelectrolytes performed better in fine particle capture and in the formation of larger flocs that resulted in a better dewaterability and less chance of overdosing. The dosing sequence in dual polyelectrolytes also affected the sludge dewaterability. Reaction mechanisms in dual polyelectrolyte conditioning were proposed based on enhanced flocculation of sludge particles.     

Effect of extra polymeric substance removal on sludge reduction potential of Bacillus licheniformis at its optimised pH condition

The effect of extracellular polymeric substances (EPSs) on the solubilisation of municipal wastewater sludge by protease‐secreting bacteria was investigated. The untreated sludge is an environmental menace, so sludge management techniques are introduced to treat and dispose sludge in a proper way. Sludge pretreatment is needed to increase the solubilisation and the efficiency of treatment methods. The present study deals with sludge solubilisation by bacterial enzymatic pretreatment. The EPS present in the sludge was removed by sodium hydroxide and formaldehyde to enhance solubilisation and increase the bacterial enzymatic activity. After the removal of EPS with different normalities of NaOH, there was about 11–18% sludge solubilisation and about 8–16% sludge reduction during bacterial pretreatment. Bacterial enzymatic pretreated sludge was further treated using aerobic digesters. From the results obtained in aerobic digestion, 0.06 N NaOH was chosen as the optimal concentration to remove EPSs.