Membrane biofouling by extracellular polymeric substances or soluble microbial products from membrane bioreactor sludge

This study extracted the soluble microbial products and loosely bound and tightly bound extracellular polymeric substances (EPS) from suspended sludge from a membrane bioreactor, original and aerobically/anaerobically digested, and compared their fouling potentials on a microfiltration membrane. The resistance of cake layer accounts for 95–98% of the total filtration resistances when filtering the whole sludges, with anaerobically digested sludge presenting the highest resistance among the three tested sludges. The tightly bound EPS has the highest potential to foul the membrane; however, the loosely bound EPS contribute most of the filtration resistances of the whole sludges. The foulants corresponding to the irreversible fouling have chemical fingerprints similar to those from loosely bound EPS, which have a greater predilection to proteins and humic substances than to polysaccharides.     

Characteristics of flux and gel layer on microfilter and non-woven fabric filter surface based on anoxic–aerobic MBRs

Non-woven fabric filter- (NWFF) and microfilter-MBR modules were made using 100?μm polypropylene and 0.25?μm polyethylene materials, respectively. The performances and mechanisms of the two processes were investigated, including additional batch filtration tests to find the function of the dynamic gel layer on the membrane surface. The HRT of both MBRs was 9?h and the operating permeate flux was 13?L/m(2)/h. The two MBRs consisted of an anoxic and aerobic reactor. The NWFF or microfilter (MF) was submerged in each of the aerobic reactors. The two MBRs showed similar performances for the removal of organic matters, suspended solids and nitrogen. Cake formation on the NWFF contributed to major resistance, while the gel layer on the microfilter or internal fouling of the pores played a key role in the fouling of the membrane surface. The amount of soluble extracellular polymer substances (EPS) (13?mg/L) of the attached sludge on the NWFF surface was larger than that (11?mg/L) of that suspended sludge. Consequently, the functional gel layer for the coarse and microfilter is established based on the relationship among the EPS, transmembrane pressure and MLSS.     

New insights into membrane fouling in a submerged anaerobic membrane bioreactor based on characterization of cake sludge and bulk sludge

A laboratory-scale submerged anaerobic membrane bioreactor (SAnMBR) treating thermomechanical pulping whitewater was operated for over 7 months to investigate and compare the characteristics of cake sludge and bulk sludge during stable state operation period. Serial analysis showed that cake sludge had a smaller particle size distribution (PSD), much higher specific filtration resistance (1.34 × 10¹⁴ m/kg), 1.5 times higher bound EPS and significantly different microbial community as compared with bulk sludge. Further analysis indicated that small flocs, bound EPS and inorganic materials play important role in cake formation process. The formed cake layer was found to have a heterogeneous structure. The results obtained in this study indicated that cake formation process started from attachment of small flocs and/or specific bacterial clusters which colonize the surface of the membrane and provide enhanced conditions that allow for cake formation to progress.     

Membrane fouling in a membrane bioreactor (MBR): sludge cake formation and fouling characteristics

A submerged membrane bioreactor (MBR) with a working volume of 1.4 L and a hollow fiber microfiltration membrane was used to treat a contaminated raw water supply at a short hydraulic retention time (HRT) of approximately 1 h. Filtration flux tests were conducted regularly on the membrane to determine various fouling resistances, and confocal laser scanning microscopy (CLSM) and scanning electron microscopy (SEM) were employed to characterize the biofouling development and sludge cake formation on the membrane. The experimental results demonstrate that the MBR is highly effective in drinking water treatment for the removal of organic pollutants, ammonia, and UV absorbance. During the MBR operation, the fouling materials were not uniformly distributed on the entire surface of all of the membrane fibers. The membrane was covered partially by a static sludge cake that could not be removed by the shear force of aeration, and partially by a thin sludge film that was frequently washed away by aeration turbulence. The filtration resistance coefficients were 308.4 x 10(11) m(-1) on average for the sludge cake, 32.5 x 10(11) m(-1) on average for the dynamic sludge film, and increased from 10.5 x 10(11) to 59.7 x 10(11) m(-1) for the membrane pore fouling after 10 weeks of MBR operation at a filtration flux of 0.5 m3/m2 x d. Polysaccharides and other biopolymers were found to accumulate on the membrane, and hence decreased membrane permeability. More important, the adsorption of biopolymers on the membrane modified its surface property and led to easier biomass attachment and tighter sludge cake deposition, which resulted in a progressive sludge cake growth and serious membrane fouling. The sludge cake coverage on the membrane can be minimized by the separation, with adequate space, of the membrane filters, to which sufficient aeration turbulence can then be applied.     

Correlating membrane fouling with sludge characteristics in membrane bioreactors: an especial interest in EPS and sludge morphology analysis

Two submerged membrane bioreactors were operated for a period of 3 months to study the filtration behavior of normal sludge and bulking sludge. Comparison of sludge morphology and bound extracellular polymeric substances (EPS) from the two systems was made to elucidate the different filtration characteristics. Experimental results showed that the membrane fouling behavior induced by bulking sludge was more severe than normal sludge. Concomitantly, the adsorption tests and atomic force microscopy observation confirmed that the EPS properties played an important role in membrane adsorption, eventually causing the different fouling behavior. Correlations between image analysis information and diluted sludge volume index (DSVI) have been identified. The combinations of EFLI/FAI (the ratio between extended filamentous lengths and floc area), from factor and floc elongation related parameters (aspect ratio or roundness) were the preferred input candidates in autoregressive exogenous model to describe the filamentous bulking phenomena, which aided in predicting membrane fouling.     

Membrane fouling properties under different filtration modes in a submerged membrane bioreactor

Four flat-sheet membrane modules, which were operated under four different filtration modes but with the same treatment capacity, were used to treat synthetic wastewater in a submerged membrane bioreactor (MBR). Particle size distribution (PSD), gel filtration chromatography (GFC), capillary suction time (CST), and three-dimensional excitation–emission matrix (EEM) fluorescence spectroscopy were used to characterize membrane fouling properties. The high instantaneous flux induced faster fouling rate and continuous filtration mode was the most applicable filtration mode in this study. The average particle size of all foulants was smaller than that of bulk sludge; and the higher the instantaneous flux was adopted, the larger the average particle size of foulants would be. Only macromolecule substances were detected in all the foulants. The macromolecule substances in the influent were degraded by microorganism and retained by membrane, and small molecular substances could pass through membrane pores to enter the effluent. The membrane foulants had poorer dewaterability compared to the mixed liquor confirmed by CST measurement. Although there were several peaks associated with protein-like fluorophores, fulvic acid-like substances and humic acid-like organics in soluble microbial products (SMP) and extracellular polymeric substances (EPS) sample, it was found that the dominant fluorescence substances in membrane foulants were protein-like substances.     

Start-Up of an Anaerobic Dynamic Membrane Digester for Waste Activated Sludge Digestion: Temporal Variations in Microbial Communities

An anaerobic dynamic membrane digester (ADMD) was developed to digest waste sludge, and pyrosequencing was used to analyze the variations of the bacterial and archaeal communities during the start-up. Results showed that bacterial community richness decreased and then increased over time, while bacterial diversity remained almost the same during the start-up. Proteobacteria and Bacteroidetes were the major phyla. At the class level, Betaproteobacteria was the most abundant at the end of start-up, followed by Sphingobacteria. In the archaeal community, richness and diversity peaked at the end of the start-up stage. Principle component and cluster analyses demonstrated that archaeal consortia experienced a distinct shift and became stable after day 38. Methanomicrobiales and Methanosarcinales were the two predominant orders. Further investigations indicated that Methanolinea and Methanosaeta were responsible for methane production in the ADMD system. Hydrogenotrophic pathways might prevail over acetoclastic means for methanogenesis during the start-up, supported by specific methanogenic activity tests.     

Microbial communities in an anaerobic dynamic membrane bioreactor (AnDMBR) for municipal wastewater treatment: Comparison of bulk sludge and cake layer

454 high-throughput pyrosequencing was employed to investigate microbial communities in bulk sludge (BS) and cake layer (CL) in an anaerobic dynamic membrane bioreactor (AnDMBR). Pyrosequencing produced 7402 and 7026 high-quality V1–V3 tags of the 16S rRNA-gene for the BS and CL samples, and the number of total OTUs estimated by Chao1 was 2906 (BS) and 2532 (CL) at a 3% distance. Phylogenetic spectrum and Venn diagrams revealed that the bacterial phyla in the two samples were nearly identical, and that the predominant microorganisms of them exhibited a high homology. Nevertheless, population distribution was different between them. Proteobacteria were the most dominant in the BS, while Bacteroidetes, Firmicutes and Spirochaetes were highly enriched in the CL, accounting for 57.6% of the total microbes. The main microbial groups in the CL were highly functionally organized. Bacteroides, Clostridia and Delta-proteobacteria were likely the pioneers for surface colonization on membranes in the AnDMBR.     

Extraction of extracellular polymeric substances (EPS) from anaerobic granular sludges: comparison of chemical and physical extraction protocols

The characteristics of the extracellular polymeric substances (EPS) extracted with nine different extraction protocols from four different types of anaerobic granular sludge were studied. The efficiency of four physical (sonication, heating, cationic exchange resin (CER), and CER associated with sonication) and four chemical (ethylenediaminetetraacetic acid, ethanol, formaldehyde combined with heating, or NaOH) EPS extraction methods was compared to a control extraction protocols (i.e., centrifugation). The nucleic acid content and the protein/polysaccharide ratio of the EPS extracted show that the extraction does not induce abnormal cellular lysis. Chemical extraction protocols give the highest EPS extraction yields (calculated by the mass ratio between sludges and EPS dry weight (DW)). Infrared analyses as well as an extraction yield over 100% or organic carbon content over 1 g g⁻¹ of DW revealed, nevertheless, a carry-over of the chemical extractants into the EPS extracts. The EPS of the anaerobic granular sludges investigated are predominantly composed of humic-like substances, proteins, and polysaccharides. The EPS content in each biochemical compound varies depending on the sludge type and extraction technique used. Some extraction techniques lead to a slightly preferential extraction of some EPS compounds, e.g., CER gives a higher protein yield.     

Step-by-step strategy for protein enrichment and proteome characterisation of extracellular polymeric substances in wastewater treatment systems

Extracellular polymeric substances (EPS) are keys in biomass aggregation and settleability in wastewater treatment systems. In membrane bioreactors (MBR), EPS are an important factor as they are considered to be largely responsible for membrane fouling. Proteins were shown to be the major component of EPS produced by activated sludge and to be correlated with the properties of the sludge, like settling, hydrophobicity and cell aggregation. Previous EPS proteomic studies of activated sludge revealed several problems, like the interference of other EPS molecules in protein analysis. In this study, a successful strategy was outlined to identify the proteins from soluble and bound EPS extracted from activated sludge of a lab-scale MBR. EPS samples were first subjected to pre-concentration through lyophilisation, centrifugal ultrafiltration or concentration with a dialysis membrane coated by a highly absorbent powder of polyacrylate-polyalcohol, preceded or not by a dialysis step. The highest protein concentration factors were achieved with the highly absorbent powder method without previous dialysis step. Four protein precipitation methods were then tested: acetone, trichloroacetic acid (TCA), perchloric acid and a commercial kit. Protein profiles were compared in 4–12 % sodium dodecyl sulphate polyacrylamide gel electrophoresis gels. Both acetone and TCA should be applied for the highest coverage for soluble EPS proteins, whereas TCA was the best method for bound EPS proteins. All visible bands of selected profiles were subjected to mass spectrometry analysis. A high number of proteins (25–32 for soluble EPS and 17 for bound EPS) were identified. As a conclusion of this study, a workflow is proposed for the successful proteome characterisation of soluble and bound EPS from activated sludge samples.     

Characterization and fouling properties of exopolysaccharide produced by Klebsiella oxytoca

Klebsiella oxytoca produced a type of exopolysaccharide (EPS) with the average molecular weight (Mw) of 116,018 Da and the average size of 260 nm. The EPS monosaccharide components contained rhamnose, fucose, arabinose, xylose, mannose, galactose and glucose and the molar ratio among them was 0.033:0.0411:0.0147:0.0051:0.2393:0.0986:0.1304. Typical EPS absorption peaks in FT-IR spectrum and pseudoplastic properties were also revealed. The polyvinylidenefluoride (PVDF) membrane showed a relatively larger flux decline resulted from the EPS fouling. The EPS filtration was dominated by more than one mechanism at the beginning phase and mainly by the cake formation at the later phase for both membranes. The pore blocking resistance had a predominant contribution to the filtration resistance and the cake resistance played a secondary role for both the membranes. The EPS adsorption resulted in a weak membrane fouling. The PVDF membrane exhibited a larger adsorption resistance than the polypropylene (PP) membrane.     

The critical flux method for reduced filter membrane fouling when monitoring high‐solids digesters

Membrane fouling currently makes filtration of high‐solids anaerobic sludges difficult and this is discouraging online monitoring of volatile fatty acids and control of high‐solids digesters. The present study tests the critical flux approach to reduce membrane fouling. Filtration tests are performed on two sludges, filtered via a side‐stream off two full‐scale digesters. Sub‐critical flux operating conditions (for minimal cake layer formation) are identified for each of the sludges and the filtration units are operated at these conditions to assess longer term performance. Results for one of the sludges (co‐digested primary and secondary sludge) is found to be encouraging, showing that sufficient flux rates (up to 40 L m^?2 h^?1) can be readily sustained to allow longer term digester monitoring and control. Filtration performance for this sludge did not deteriorate significantly over the test period. Results for the other test sludge (digested thermally hydrolyzed waste activated sludge) were not as favorable and indicated that application may be limited for very high solids digesters (>5% total solids concentration). Differences in filtration behavior for the two test sludges were ascribed to the presence of complex soluble organics, the concentration of sludge solids and their particle size. © 2013 American Institute of Chemical Engineers Biotechnol. Prog., 29:1059–1063, 2013     

Treatment of wastewater from abattoirs before land application--a review

Pre-treatments are screening, catch basins, flotation, equalization, and settlers for recovering proteins and fats from abattoir wastewater. With chemical addition, dissolved air flotation (DAF) units can achieve chemical oxygen demand (COD) reductions ranging from 32% to 90% and are capable of removing large amounts of nutrients. Aerobic trickling towers reduced soluble COD by additional 27% but did not reduced total COD. Chemical-DAF reduced 67% of total COD and soluble COD. About 40-60% of the solids or approximately 25-35% of the biological oxygen demand (BOD) load can be separated by pre-treatment screening and sedimentation. Anaerobic systems are lagoon, anaerobic contact (AC), up-flow anaerobic sludge blanket (UASB), anaerobic sequence batch reactor (ASBR), and anaerobic filter (AF) processes. Abattoir wastewater is well suited to anaerobic treatment because it is high in organic compounds. Typical reductions of up to 97% BOD, 95% SS and 96% COD are reported. UASB's average COD removal efficiencies are of 80-85%. UASB seems to be a suitable process for the treatment of abattoir wastewater, due to its ability to maintain a sufficient amount of viable sludge. Wastewater in abattoirs can be reduced by treatment of immersion chiller effluent by membrane filtration which can produce recyclable water. Total organic C can be reduced below 100mg/L, and bacteria can not pass through the membrane pores. The abattoir waste minimization options are also discussed.     

Formation of extracellular polymeric substances from acidogenic sludge in H<Subscript>2</Subscript>-producing process

In this study, the formation of extracellular polymeric substances (EPS) and surface characteristics of an acidogenic sludge in anaerobic H₂-producing process was investigated. Results show that carbohydrates, proteins, and humic substances were the dominant components in bound EPS (BEPS), while in soluble EPS (SEPS), carbohydrates were the main component. The total content of BEPS initially increased but then kept almost unchanged during fermentation from 25 to 35 h; after that, it slightly decreased. The total content of SEPS increased to 172.5 ± 0.05 mg C g⁻¹ volatile suspended solid with the time that increased to 23.5 h, and then rapidly decreased until 43 h; thereafter, it kept almost unchanged. The SEPS had good correlations with the specific H₂ production rate, substrate degradation rate, and specific aqueous products formation rate, but the BEPS seemed to have no such correlations with these specific rates. Results also confirm that part of EPS could be utilized by the H₂-producing sludge. As the substrate was in short supply, the EPS would be hydrolyzed to sever as carbon and energy source.     

Influence of elevated pH shocks on the performance of a submerged anaerobic membrane bioreactor

The effects of elevated pH shocks on the performance and membrane fouling of a submerged anaerobic membrane bioreactor (SAnMBR) treating thermomechanical pulping (TMP) whitewater was studied over a 120-day period. Changes in chemical oxygen demand (COD) removal, biogas production, sludge and cake layer properties, and their correlations to membrane fouling – before and after pH shocks – were systematically studied using various analytical tools. The results showed that a pH 8.0 shock had a minor impact, while pH 9.1 and 10.0 shocks exerted significant long-lasting negative impacts on COD removal, biogas production and membrane filtration performance of the SAnMBR. When the normal pH (7.0) was resumed, it took approximately 1, 6, and 30 days for the performance to recover for pH 8.0, 9.1 and 10.0 shocks, respectively. The elevated pH shocks induced the dispersion of sludge flocs and resulted in the accumulation of colloids and solutes or biopolymers in the sludge suspension, and thus deteriorated membrane performance. Statistical analysis showed that the ratio of proteins (PN) to polysaccharides (PS) in extracellular polymeric substances (EPS) had a strongly negative effect on the membrane fouling rate. There were smaller size particles deposited on the membrane surface and a more compact and denser cake layer was formed after being exposed to an alkaline shock at pH 10, resulting in higher membrane fouling rates.     

Changes in the composition of extracellular polymeric substances in activated sludge during anaerobic storage

Changes in the chemical composition of organic compounds in total activated sludge, activated sludge extracellular polymeric substances (EPS), and sludge bulk water during anaerobic storage (12 days) were studied. The background for the study was that anaerobic storage of activated sludge, which often takes place at wastewater treatment plants before dewatering, causes a deterioration of the dewaterability. The reasons are not known at present, but may be related to changes in exopolymer composition of the flocs. The results showed that a fast decrease in total sludge protein and carbohydrate took place within 3 days of anaerobic storage as a result of degradation processes, which accounted for approximately 20% of the organic fraction. The amount of uronic acids and humic compounds remained almost constant in the sludge. The EPS were extracted from the floc matrix using a cationexchange resin. In the EPS matrix a similar initial (2–3 days) degradation of proteins and carbohydrate took place, whereas the content of DNA and uronic acids showed minor changes. The extractability of humic compounds increased during the first 3 days of storage. No changes in extractability of the carbohydrate were observed. A fraction of the EPS protein was found to be difficult to extract but was observed to be degraded during the anaerobic storage. The EPS composition was further characterized by high-performance size-exclusion chromatography analysis obtained by on-line UV detection and post-column detection of proteins, carbohydrates, humic compounds and DNA. Four fractions of polysaccharides were found, of which only one was responsible for the decrease in the carbohydrate content observed with storage time. The fraction was presumably of low molecular mass. Humic compounds and volatile fatty acids (acetate and propionate) were released to the bulk water from the flocs during the storage. A possible mechanism to explain the reduced dewaterability developed during anaerobic storage, partly because of the observed changes in EPS, is discussed.     

Soluble microbial products (SMP) and soluble extracellular polymeric substances (EPS) from wastewater sludge

Laspidou and Rittmann (Water Research 36:2711–2720, 2002) proposed that the soluble extracellular polymeric substances (EPS) are identical to soluble microbial products (SMP) in sludge liquor. In this paper, we compared the physicochemical characteristics of the SMP and soluble EPS from original and aerobically or anaerobically digested wastewater sludge. The surface charges, particle sizes, residual turbidities of polyaluminum chloride (PACl) coagulated supernatant, and chemical compositions of the SMP and soluble EPS containing suspensions were used as comparison index. Experimental results revealed that the particles in SMP and soluble EPS fractions extracted from original wastewater sludge, before and after digestion, were not identical in all physicochemical characteristics herein measured. The current test cannot support the proposal by Laspidou and Rittmann (Water Research 36:2711–2720, 2002) that SMP is identical to the soluble EPS from a wastewater sludge.     

Bonding Form Analysis of Metals and Sulfur Fractionation in Methanol‐Grown Anaerobic Granular Sludge

This study investigates the metal and sulfur bonding form distribution in mesophilic (30 °C, pH 7) methanol‐grown anaerobic granular sludge from upflow anaerobic sludge bed reactors operating at an organic loading rate of 3.8 g CH₃OH‐COD/L d. This was achieved by applying a modified Tessier sequential extraction scheme to investigate the metal bonding forms and a sequential extraction scheme for sulfur and simultaneously extracted metals to granular sludge samples of the reactors after 0, 22, 35 and 43 days of operation. Metals were also determined in the sulfur extracts. Co and Ni predominated in their oxidizable bonding forms, which increased together with the pseudo‐total content during reactor operation. An omission of Co and Ni from the influent led to only a minor decline of the pseudo‐total content in the sludge, mainly from the acid‐soluble fraction. The ratio of simultaneously extracted metals (Co, Fe, Mn, Ni) to acid‐volatile sulfides was lower than 1, indicating that the sludge contained sufficient sulfide to bind the metals as metal monosulfides. The bioavailability of metals in the methanol‐grown anaerobic granular sludge investigated is therefore mainly controlled by sulfide formation/dissolution.     

Comparison between a moving bed membrane bioreactor and a conventional membrane bioreactor on membrane fouling

A membrane bioreactor filled with carriers instead of activated sludge named a moving bed membrane bioreactor (MBMBR) was investigated to minimize the effect of suspended solids on membrane fouling. The MBMBR and a conventional membrane bioreactor (CMBR) were operated in parallel for about two months. Unexpectedly, the rate of membrane fouling in MBMBR was about three times of that in CMBR. MBMBR showed a higher cake layer resistance than CMBR due to plenty of filamentous bacteria inhabited in suspended solids in MBMBR. Protein and polysaccharide contents of soluble EPS in MBMBR were obviously larger than those in CMBR. It could be speculated that the overgrowth of filamentous bacteria in MBMBR resulted in severe cake layer and induced a large quantity of EPS, which deteriorated the membrane fouling.     

Composition and distribution of extracellular polymeric substances in aerobic flocs and granular sludge

Extracellular polymeric substances (EPS) were quantified in flocculent and aerobic granular sludge developed in two sequencing batch reactors with the same shear force but different settling times. Several EPS extraction methods were compared to investigate how different methods affect EPS chemical characterization, and fluorescent stains were used to visualize EPS in intact samples and 20-mum cryosections. Reactor 1 (operated with a 10-min settle) enriched predominantly flocculent sludge with a sludge volume index (SVI) of 120 +/- 12 ml g(-1), and reactor 2 (2-min settle time) formed compact aerobic granules with an SVI of 50 +/- 2 ml g(-1). EPS extraction by using a cation-exchange resin showed that proteins were more dominant than polysaccharides in all samples, and the protein content was 50% more in granular EPS than flocculent EPS. NaOH and heat extraction produced a higher protein and polysaccharide content from cell lysis. In situ EPS staining of granules showed that cells and polysaccharides were localized to the outer edge of granules, whereas the center was comprised mostly of proteins. These observations confirm the chemical extraction data and indicate that granule formation and stability are dependent on a noncellular, protein core. The comparison of EPS methods explains how significant cell lysis and contamination by dead biomass leads to different and opposing conclusions.