Identification of biofoulant of membrane bioreactors in soluble microbial products

To reveal primary biofoulant in soluble microbial products (SMP) and/or soluble extracellular polymeric substances (EPS), after removal of sludge particles, activated sludge samples were subjected to microfiltration tests in a submerged MBR. Filtration resistance directly correlates with the saccharide concentration. Saccharides in wastewater from several sources contained uronic acids, which increased the filtration resistance. When the microfiltration test liquids contained saccharides over 80 mg l⁻¹, a gelatinous mass remained on the membrane surface after filtration and contained concentrations of saccharides and uronic acids 50 times higher than the original test liquid while only trace amounts of these substances were contained in the filtrate. The gelatinous mass contained high molecular weight substances of 10⁶-10⁸ Da, suggesting the presence of polysaccharides. However, molecules of this size were calculated to be much smaller than the pore size of the membrane. Ethylenediaminetetraacetic acid decreased filtration resistance, suggesting that polysaccharides containing uronic acid units could undergo intermolecular or intramolecular ionic cross-linking by polyvalent cations and form the gel, thus clogging the membrane pores as an actual biofoulant.     

Complexity of ultrafiltration membrane fouling caused by macromolecular dissolved organic compounds in secondary effluents

Recent investigations indicate the relevance of extracellular polymeric substances (EPS) in terms of fouling of low-pressure membranes in advanced wastewater treatment. In this study, the high impact of the macromolecular fraction of effluent organic matter on fouling was confirmed in cross-flow ultrafiltration experiments using secondary effluent with and without autochthonous biopolymers. A method for the extraction of a natural mixture of EPS derived from the bacterium Sinorhizobium sp. is presented. Ultrafiltration of solutions of this bacterial EPS extract revealed a correlation between the concentration of EPS and the loss of permeate flux. However, in ultrafiltration tests using extracted bacterial EPS in a model solution as well as in secondary effluent without autochthonous biopolymers, the extent of membrane fouling was not identical with the fouling provoked by secondary effluent organic matter, although the biopolymer concentrations were comparable. The differences in the fouling behaviour of the extracted bacterial EPS and effluent organic matter are considered to be due to different compositions of the biopolymer fraction in terms of proteins, polysaccharides, and other organic colloids, indicating a particular impact of proteins on ultrafiltration membrane fouling.     

A comparative study of fouling-related properties of sludge from conventional and membrane enhanced biological phosphorus removal processes

The physical and biochemical properties of activated sludge mixed liquor, including floc size distribution, zeta potential, relative hydrophobicity, and bound and unbound (soluble) extracellular polymeric substances (EPS), were examined in this study to evaluate their relationship to membrane fouling. Mixed liquors from a membrane enhanced biological phosphorus removal (MEBPR) process and a conventional enhanced biological phosphorus removal (CEBPR) process were compared. It was found that the floc size distribution and the amount of soluble EPS in the mixed liquor were the most important properties that significantly influenced the fouling propensity of sludge. Contrary to the literature, the content of EPS bound in activated sludge flocs was not found to be directly associated with membrane fouling, and sludge surface properties such as zeta potential and relative hydrophobicity were not closely related to the observed differences in the fouling tendencies of the two types of sludge.     

Extracellular polymeric substances (EPS) properties and their effects on membrane fouling in a submerged membrane bioreactor

A pilot-scale submerged membrane bioreactor (MBR) for real municipal wastewater treatment was operated for over one year in order to investigate extracellular polymeric substances (EPS) properties and their role in membrane fouling. The components and properties of bound EPS were examined by the evaluation of mean oxidation state (MOS) of organic carbons, Fourier transform infrared (FT-IR) spectroscopy, three-dimensional excitation-emission matrix (EEM) fluorescence spectroscopy, and gel filtration chromatography (GFC), etc. Test results showed that MOS of organic carbons in the bound EPS was ranging from −0.14 to −0.51, and major components could be assessed as proteins and carbohydrates. FT-IR analysis confirmed the presence of proteins and carbohydrates in the bound EPS. The organic substances with fluorescence characteristics in the bound EPS were identified as proteins, visible humic acid-like substances and fulvic acid-like substances by EEM technology. GFC analysis demonstrated that EPS had part of higher MW molecules and a broader MW distribution than the influent wastewater. It was also found that a high shear stress imposed on mixed liquor could result in the release of EPS, which would in turn influence membrane fouling in MBRs. Bound EPS solution was observed to have a stronger potential of fouling than mixed liquor. During long-term operation of the MBR, bound EPS demonstrated positive correlations with membrane fouling while temperature was verified as a negative factor affecting EPS concentration. Compared to tightly bound EPS (TB-EPS), loosely bound EPS (LB-EPS) showed more significant correlations with membrane fouling. This critical investigation would contribute towards a better understanding of the behavior, composition and fouling potential of EPS in MBR operation.     

Effect of PAC addition on sludge properties in an MBR treating high strength wastewater

This paper examines the sludge characteristics in a submerged membrane bioreactor (MBR) operated on a high strength wastewater from an alcohol distillery. Two membrane bioreactors, each with a 30 μm mesh filter, were investigated with and without addition of powdered activated carbon (PAC). Experiments were conducted with varying organic loading rates ranging from 3.4 to 6.9 kgCOD m⁻³ day⁻¹ and the specific oxygen uptake rate (SOUR), sludge volume index (SVI), mixed liquor suspended solids (MLSS), particle size and extracellular polymeric substances (EPS) were monitored over a 180 day period. Respirometric experiments did not show enhancement in microbial activity with PAC supplementation. Addition of PAC decreased the SVI thereby perceptibly improving sludge dewaterability. The sludge particle size, which increased with time, appeared to be independent of PAC addition but was influenced by the aeration intensity. PAC also did not affect the sludge EPS concentration; however, the EPS composition, in terms of protein/carbohydrate (polysaccharide) ratio was altered resulting in a high P/C ratio. FTIR analysis of the sludge samples indicated that the functional groups associated with the sludge polysaccharides appear to be involved in its interaction with PAC.     

Sludge properties and their effects on membrane fouling in submerged anaerobic membrane bioreactors (SAnMBRs)

Two submerged anaerobic membrane bioreactors (SAnMBRs) (thermophilic vs. mesophilic) were operated for a period of 3.5 months with kraft evaporator condensate at a feed chemical oxygen demand of 10,000 mg/L. The results show that the filtration behavior of the two systems was significantly different. The filtration resistance in the thermophilic SAnMBR was about 5–10 times higher than that of the mesophilic system when operated under similar hydrodynamic conditions. Comparison of sludge properties and cake layer structure from the two systems was made to elucidate major factors governing the different filtration characteristics. There were more soluble microbial products (SMP) and biopolymer clusters (BPC) produced and a larger portion of fine flocs (<15 μm) in the thermophilic SAnMBR. Analysis of bound extracellular polymeric substances (EPS) showed that the thermophilic sludge had a higher protein/polysaccharide ratio in EPS, as compared to that in the mesophilic sludge. A series of analyses, including Fourier transform infrared (FTIR) spectroscopy, energy dispersive X-ray spectroscopy (EDX), confocal laser scanning microscopy (CLSM), scanning electron microscopy (SEM), atomic force microscopy (AFM) and particle size analyzer showed that the cake layer formed in the thermophilic SAnMBR contained higher levels of both organic and inorganic foulants, smaller particle sizes, and especially, a denser and more compact sludge cake structure. These results indicate that floc size, SMP, BPC, bound EPS as well as cake layer structure are the major factors governing membrane fouling in SAnMBR systems.     

Role of extracellular polymeric substances in bioflocculation of activated sludge microorganisms under glucose-controlled conditions

Extracellular polymeric substances (EPS) secreted by suspended cultures of microorganisms from an activated sludge plant in the presence of glucose were characterized in detail using colorimetry, X-ray photoelectron spectroscopy (XPS) and Fourier transform infrared (FTIR) spectroscopy. EPS produced by the multi-species community were similar to literature reports of pure cultures in terms of functionalities with respect to C and O but differed subtly in terms of N and P. Hence, it appears that EPS produced by different microorganisms maybe homologous in major chemical constituents but may differ in minor components such as lipids and phosphodiesters. The role of specific EPS constituents on microbial aggregation was also determined. The weak tendency of microorganisms to bioflocculate during the exponential growth phase was attributed to electrostatic repulsion when EPS concentration was low and acidic in nature (higher fraction of uronic acids to total EPS) as well as reduced polymer bridging. However, during the stationary phase, polymeric interactions overwhelmed electrostatic interactions (lower fraction of uronic acids to total EPS) resulting in improved bioflocculation. More specifically, microorganisms appeared to aggregate in the presence of protein secondary structures including aggregated strands, β-sheets, α- and 3-turn helical structures. Bioflocculation was also favored by increasing O-acetylated carbohydrates and overall C-(O,N) and functionalities.     

Polymeric compounds in activated sludge supernatant -- Characterisation and retention mechanisms at a full-scale municipal membrane bioreactor

In this study, for the first time a full-scale membrane bioreactor (MBR) was investigated with focus on organic compounds in activated sludge over a period of approximately 2 years. Soluble extracellular polymeric substances (EPS) in the sludge supernatant and permeate as well as bound EPS extracted from fouled membranes were determined photospectrometrically and revealed a typical composition of three main components in the order metals>humic acids>carbohydrates>proteins. Results showed an important influence on membrane fouling by soluble humic substances and carbohydrates in complexes with metal cations. It was found that Fe(2+) and Fe(3+) play a decisive role in natural organic matter (NOM) complexation and subsequent membrane blockage. The determination of molar mass distribution in supernatant and permeate by size exclusion chromatography (SEC) revealed a significant retention of macromolecular compounds by the porous membranes in the range of 10-50%.     

Relation between EPS adherence, viscoelastic properties, and MBR operation: Biofouling study with QCM-D

Membrane fouling is one of the main constraints of the wide use of membrane bioreactor (MBR) technology. The biomass in MBR systems includes extracellular polymeric substances (EPS), metabolic products of active microbial secretion that adversely affect the membrane performance. Solids retention time (SRT) in the MBR is one of the most important parameters affecting membrane fouling in MBR systems, where fouling is minimized at optimal SRT. Among the operating parameters in MBR systems, SRT is known to strongly influence the ratio of proteins to polysaccharides in the EPS matrix. In this study, we have direct evidence for changes in EPS adherence and viscoelastic properties due to changes in the sludge removal rate that strongly correlate with the membrane fouling rate and EPS composition. EPS were extracted from a UF membrane in a hybrid growth MBR operated at sludge removal rates of 59, 35.4, 17.7, and 5.9 L day^-1 (corresponding SRT of 3, 5, 10, and 30 days, respectively). The EPS adherence and adsorption kinetics were carried out in a quartz crystal microbalance with dissipation monitoring (QCM-D) technology in several adsorption measurements to a gold sensor coated with Polyvinylidene Fluoride (PVDF). EPS adsorption to the sensor surface is characterized by a decrease of the oscillation frequency and an increase in the dissipation energy of the sensor during parallel flow of aqueous media, supplemented with EPS, above the sensor surface. The results from these experiments were further modeled using the Voigt based model, in which the thickness, shear modulus, and shear viscosity values of the adsorbed EPS layers on the PVDF crystal were calculated. The observations in the QCM-D suggested that the elevated fouling of the UF membrane is due to higher adherence of the EPS as well as reduction in viscosity and elasticity of the EPS adsorbed layer and elevation of the EPS fluidity. These results corroborate with confocal laser scanning microscopy (CLSM) image analysis showing thicker EPS in close proximity to the membrane surface operated at reactor conditions which induced more fouling at elevated sludge removal rates.     

A comparison of membrane fouling under constant and variable organic loadings in submerge membrane bioreactors

The aim of this study is to compare the effect of constant and variable influent organic loadings on membrane fouling in submerged membrane bioreactors (sMBRs). Two identical lab-scale sMBRs were operated for 162 days at an SRT of 30 days, whereas the influent organic loading was kept constant in one MBR, and varied in another. The microbial characteristics of sludge in terms of MLSS, bound EPS, EPS in the supernatant and particle size distribution were investigated in order to evaluate their respective effect on membrane fouling. During the start-up period, membrane fouling in the MBR fed with variable loadings was more serious than that in the MBR with the constant loading. However, at the stable state, the fouling tendency was clearly reversed with less membrane fouling for variable feed strength. It was shown that the contents of polysaccharides in the supernatant and particle size of the bioflocs were responsible for the observed differences in the fouling tendencies of the two MBRs.     

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.     

Impact of colloidal and soluble organic material on membrane performance in membrane bioreactors for municipal wastewater treatment

Two parallel membrane bioreactors (2 m3 each) were operated over a period of 2 years. Both pilots were optimised for nitrification, denitrification, and enhanced biological phosphorous elimination, treating identical municipal wastewater under comparable operating conditions. The only constructional difference between the pilots was the position of the denitrification zone (pre-denitrification in pilot 1 and post-denitrification in pilot 2). Despite identical modules and conditions, the two MBRs showed different permeabilities and fouling rates. The differences were not related to the denitrification scheme. In order to find an explanation for the different membrane performances, a one-year investigation was initiated and the membrane performance as well as the operating regime and characteristics of the activated sludge were closely studied. MLSS concentrations, solid retention time, loading rates, and filtration flux were found not to be responsible for the different performance of the submerged modules. These parameters were kept identical in the two pilot plants. Instead, the non-settable fraction of the sludges (soluble and colloidal material, i.e. polysaccharides, proteins and organic colloids) was found to impact fouling and to cause the difference in membrane performance between the two MBR. This fraction was analysed by spectrophotometric and size exclusion chromatography (SEC) methods. In a second step, the origin of these substances was investigated. The results point to microbiologically produced substances such as extracellular polymeric substances (EPS) or soluble microbial products (SMP).     

The influence of aeration intensity on predation and EPS production in membrane bioreactors

Shear, in the form of vigorous aeration, is used to control fouling in membrane bioreactor (MBR) systems. However, shear also influences the physicochemical and biological properties of MBR biomass. The current study examines the relationship between the aeration intensity and extracellular polymeric substance (EPS) production in MBRs. Two identical submerged MBRs were operated in parallel but the aeration rate was three times greater in one of the MBRs. The concentrations of floc-associated and soluble EPS were monitored for the duration of the experiment. Microscopic images and floc-size measurements were also collected regularly. The membrane fouling potential of the biomass was quantified using the flux-step method. Increased aeration did not have a direct effect on soluble or floc-associated EPS production in the microfiltration MBRs. However, aeration intensity had a significant effect on predatory organisms. Large aquatic earthworms, Aeolosoma hemprichi, proliferated under lower shear conditions but were never observed in the high shear reactor. Predation by A. hemprichi resulted in increased floc-associated and soluble EPS production. Thus, the mixing conditions in the low shear MBR indirectly resulted in increased soluble EPS concentrations and higher fouling potential. This research suggests that predation can have a significant impact on the production rates of floc-associated and soluble EPS - key parameters driving membrane fouling in MBRs.     

Effect of temperature shocks on membrane fouling in membrane bioreactors

Temperature is known to influence the biological performance of conventional activated sludge systems. In membrane bioreactors (MBRs), temperature not only affects the bioconversion process but is also shown to have an effect on the membrane performance. Four phenomena are generally reported to explain the higher resistance for membrane filtration found at lower temperatures: (1) increased mixed liquor viscosity, reducing the shear stress generated by coarse bubbles, (2) intensified deflocculation, reducing biomass floc size and releasing EPS into the mixed liquor, (3) lower backtransport velocity and (4) reduced biodegradation of COD. Although the higher resistance at low temperatures has been reported in several papers, the relation with supernatant composition has not been investigated before. In this paper, the composition of the soluble fraction of the mixed liquor is related to membrane performance after exposing the sludge to temperature shocks. Flux step experiments were performed in an experimental system at 7, 15, and 25° Celsius with sludge that was continuously recirculated from a pilot-scale MBR. After correcting the permeate viscosity for temperature, higher membrane fouling rates were obtained for the lower temperature in combination with low fouling reversibility. The soluble fraction of the MBR mixed liquor was analysed for polysaccharides, proteins and submicron particle size distribution. At low temperature, a high polysaccharide concentration was found in the experimental system as compared to the MBR pilot. Upon decreasing the temperature of the mixed liquor, a shift was found in particle size towards smaller particles. These results show that the release of polysaccharides and/or submicron particles from sludge flocs could explain the increased membrane fouling at low temperatures.     

Comparison of sludge characteristics and performance of a submerged membrane bioreactor and an activated sludge process at high solids retention time

This work aims to compare biomass structure and performance of a submerged membrane bioreactor (SMBR) and an activated sludge process (ASP) treating the same domestic wastewater. The influence of the separation technique (membrane filtration or settling) and operation at high sludge-retention time (SRT) were investigated. Over the entire range of SRT (10-110 days), the SMBR achieved very good organic removal efficiencies, ranging from 90.8+/-0.2% to 94.2+/-1.6% based on total COD (TCOD), whereas those of ASP were between 87.4+/-1.8% and 90.3+/-0.8%. The contribution of the membrane in the increase in performance was due to total suspended solid retention and also partly due to retention of proteins and polysaccharides of the sludge supernatant. No significant difference in excess sludge production was observed between the two processes operated at the same SRT, but sludge production in SMBR decreased from 0.31 to 0.13 g(VSS)g(COD)(-1) as SRT increased from 9 to 110 days. The difference in sludge characteristics and performance was especially pronounced as SRT increased, resulting in deterioration of sludge settleability and effluent quality of the ASP (filamentous bacteria, increase of protein and polysaccharide release). Membrane filtration induced accumulation of soluble and colloidal proteins and polysaccharides which were progressively degraded in the supernatant as the SRT increased. At similar SRT, no significant difference was observed in the amount of extractable exocellular polymeric substances (bound EPS) from ASP and SMBR sludge. However as the SRT increased, the total specific amount of bound EPS in flocs decreased and the ratio proteins/polysaccharides also decreased. Concomitantly, laser diffraction analysis, microscopic observations, turbidity and DSVI measurement showed that the SRT increase induced significant modifications in sludge morphology in SMBR: decrease in floc size, densification of aggregates, and development of non-flocculating organisms.     

Characterization of biofilm structure and its effect on membrane permeability in MBR for dye wastewater treatment

Two membrane bioreactors were operated at aerobic (DO=6.0mg/L) and anoxic (DO<0.3mg/L) conditions for the treatment of synthetic dye wastewater to determine the effect of dissolved oxygen on membrane filterability. The rate of membrane fouling for the anoxic MBR was five times faster than that for the aerobic MBR. Differences in the nature of the biofilm that was formed on the membrane surface as the result of different DO level was the main factor in the different fouling rates. The biofilm structure was characterized using digital image analysis techniques. Biofilm images were obtained using confocal laser scanning microscopy (CLSM) at various operation points. Structural parameters were then computed from these images using an image analysis software (ISA-2). The structural parameters indicated that the anoxic biofilm was thinner than the aerobic biofilm but the anoxic biofilm was spread out on the membrane surface more uniformly and densely, resulting in the higher membrane fouling. Based on the extracellular polymeric substances (EPS) visualization and quantification, it was also found that EPS, key membrane foulants were spread out more uniformly in the anoxic biofilm in spite of lower amount of EPS compared to that in the aerobic biofilm.     

Influence of shear on the production of extracellular polymeric substances in membrane bioreactors

Shear is used to control fouling in membrane bioreactor (MBR) systems. However, shear also influences the physicochemical and biological properties of MBR biomass. The current study examines the relationship between the level of shear and extracellular polymeric substance (EPS) production in MBRs. Two identical MBRs were operated in parallel where the biomass in one reactor was exposed to seven times greater shear forces. The concentrations of floc-associated and soluble EPS were monitored for the duration of the experiment. The stickiness of extracted floc-associated EPS from each reactor was also characterized using atomic force microscopy. A mathematical model of floc-associated and soluble EPS production was applied to quantitatively describe changes in EPS production with shear. Biomass grown in a high shear environment has lower floc-associated EPS production compared to biomass grown in a lower shear environment. This decrease in floc-associated EPS production also corresponds to a decrease in soluble EPS production, which can be explained by both the lower concentration of floc-associated EPS and the production of stickier floc-associated EPS that is more erosion resistant in the high shear reactor. This research suggests that mechanical stresses can have a significant impact on the production rates of floc-associated and soluble EPS—key parameters governing membrane fouling in MBRs.     

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.     

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).     

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.