Amount Of Extracellular Polymeric Substances Research Articles

The short- and long-term effects of Mn2+ on biogranule-based anaerobic ammonium oxidation (anammox)

The short- and long-term effects of Mn2+ on the performance of anaerobic ammonium oxidation (anammox) granules were investigated in the present study. Anammox activity was determined at various Mn2+ concentrations in batch assays, and the 50% inhibition concentration value was determined to be 7.33mgL−1. However, no obvious deterioration of reactor performance was observed during the 150-day continuous-flow operation, and the nitrogen removal efficiency of the test reactor (R1) fluctuated between 91% and 92% as the Mn2+ concentration was increased from 1 to 200mgL−1. Additionally, the specific anammox activity, heme c content and the amounts of extracellular polymeric substances in the anammox biomass increased and then subsequently decreased. The results demonstrated that short-term exposure to Mn2+ has a negative effect on anammox biomass, but the biomass could tolerate Mn2+ stress after acclimation to a high concentration of 200mgL−1 at the end of the continuous-flow experiment.

The changes of proteins and polysaccharides in extracellular polymeric substance for Spirogyra fluviatilis under different salinity

Spirogyra is a genus of widely distributed, large green fresh water algae. This study discovered that changes in salinity can induce Spirogyra fluviatilis to produce amounts of extracellular polymeric substance (EPS) when controlling other environmental conditions. If culturing S. fluviatilis with salinity greater than a 3.0‰ medium for 4 hours, the secretion EPS will be changed. And the level of polysaccharides and proteins, the primary components of EPS, is slightly increased in accordance with the increase in the salinity. But the proteins to polysaccharides ratio changes are not significantly

Comparison of two chemical extraction methods for proteins and polysaccharides of Spirogyra fluviatilis in extracellular polymeric substances

Under natural circumstances, Spirogyra fluviatilis excretes large amounts of extracellular polymeric substances (EPS) and forms a mucilage layer. This study used two algal EPS extraction methods such as NH4OH and formaldehyde-NaOH to compare the extraction yields of EPS and the levels of its primary components of polysaccharides and proteins. The results indicated that the extraction yields of EPS using formaldehyde-NaOH and NH4OH are 8.05 mg EPS/g-fresh algae and 5.26 mg EPS/g-fresh algae, respectively. The ratio of proteins to polysaccharides in EPS is around 0.24 to 0.34, which is not a significant difference between the two extraction methods. In other words, the level of polysaccharides is always higher than the level of proteins in EPS extraction with both two methods

Effects of single and combined copper/perfluorooctane sulfonate on sequencing batch reactor process and microbial community in activated sludge

Long-term exposure experiments with single and combined pollutants of copper (Cu)/perfluorooctane sulfonate (PFOS) were conducted to explore the influence on activated sludge in SBRs. Compared with the control, the removal of organics, nitrogen and phosphorus in the presence of PFOS exhibited no apparent difference, but reduced in different degrees when Cu and Cu/PFOS existed. PFOS exposure deteriorated the settling performance of activated sludge with SVI value and amount of extracellular polymeric substance (EPS) increasing, but posed little impacts on microbial activity (dehydrogenase, protease) and antioxidant activity (SOD, CAT). Under Cu and Cu/PFOS loading, dehydrogenase and protease activity were observed to decrease as well as SOD and CAT activity. The sequencing results revealed that bacterial richness and community diversity reduced under Cu and Cu/PFOS exposure. Overall, adverse effect of combined pollution was lower than that of single Cu in long-time due to antagonistic effect existed between Cu and PFOS.

Different Types of Diatom-Derived Extracellular Polymeric Substances Drive Changes in Heterotrophic Bacterial Communities from Intertidal Sediments.

Intertidal areas support extensive diatom-rich biofilms. Such microphytobenthic (MPB) diatoms exude large quantities of extracellular polymeric substances (EPS) comprising polysaccharides, glycoproteins and other biopolymers, which represent a substantial carbon pool. However, degradation rates of different EPS components, and how they shape heterotrophic communities in sediments, are not well understood. An aerobic mudflat-sediment slurry experiment was performed in the dark with two different EPS carbon sources from a diatom-dominated biofilm: colloidal EPS (cEPS) and the more complex hot-bicarbonate-extracted EPS. Degradation rate constants determined over 9 days for three sediment fractions [dissolved organic carbon (DOC), total carbohydrates (TCHO), and (cEPS)] were generally higher in the colloidal-EPS slurries (0.105–0.123 d−1) compared with the hot-bicarbonate-extracted-EPS slurries (0.060–0.096 d−1). Addition of hot-bicarbonate-EPS resulted in large increases in dissolved nitrogen and phosphorous by the end of the experiment, indicating that the more complex EPS is an important source of regenerated inorganic nutrients. Microbial biomass increased ~4–6-fold over 9 days, and pyrosequencing of bacterial 16S rRNA genes revealed that the addition of both types of EPS greatly altered the bacterial community composition (from 0 to 9 days) compared to a control with no added EPS. Bacteroidetes (especially Tenacibaculum) and Verrucomicrobia increased significantly in relative abundance in both the hot-bicarbonate-EPS and colloidal-EPS treatments. These differential effects of EPS fractions on carbon-loss rates, nutrient regeneration and microbial community assembly improve our understanding of coastal-sediment carbon cycling and demonstrate the importance of diverse microbiota in processing this abundant pool of organic carbon.

Anthropogenically driven habitat formation by a tube dwelling diatom on the Northern Patagonian Atlantic coast

The tube dwelling diatom Berkeleya rutilans (Trentepohl) Grunow plays a key role as early colonizer and bloom former in coastal zones. Exuding large quantities of extracellular polymeric substances (EPS), it can form dense colonies in mucilaginous macroscopic branches, containing thousands of cells. Due to their pronounced three dimensional growths of its mucilaginous structures, it supports a variety of organisms and traps grains and detritus, which makes it an important habitat former and ecosystem engineer, contributing to sediment stabilization, which is a crucial issue in sedimentary areas. In the present study we investigated the identity and structural morphology of B. rutilans, blooming in a tidal channel in Northern Patagonia (S40° 43′ W64° 56′) and experimentally tested its potential physiological responses (e.g. growth rate) to nutrient elevation. The observed morphological plasticity and measured high growth rates under nutrient exposure make B. rutilans a likely indicator for eutrophication in sedimentary marine habitats. As to our knowledge the present study provides the first record of B. rutilans for Argentinean waters, we discussed the potential reasons for its occurrence and evaluated the ecological impacts of its presence. Due to the observed high colonization capability and rapid response to environmental alterations (e.g. eutrophication, substrate changes) it seems to benefit from human activities, which will consequently favor its further expansion within the disturbed area.

Removal of nitrogen from wastewaters by anaerobic ammonium oxidation (ANAMMOX) using granules in upflow reactors

Nitrogen pollution of waters has sometimes caused severe eutrophication, leading to the death of fishes and most aquatic life. There is therefore a need for efficient and cost-effective methods to remove nitrogen from ammonium-rich wastewaters. Anaerobic ammonium oxidation (ANAMMOX) is a promising process to remove nitrogen because this process directly oxidizes ammonium (NH4 +) to dinitrogen gas (N2) under anoxic condition. Nonetheless, a challenge of this process is that chemolithoautotrophic Anammox bacteria grow slowly at the beginning, thus resulting in low Anammox biomass and instability of reactors. Such issues can be overcome by granulation of the Anammox sludge. Here, we review the characteristics of the Anammox bacteria, and the formation, structure and flotation of Anammox granules under high hydraulic loadings. We also evaluate the performances of full-scale granular Anammox processes. The major points are: 1) Anammox bacteria secrete a large amount of extracellular polymeric substances (EPS), up to 415 mg g−1 of volatile suspended solids (VSS), containing many hydrophobic functional groups that facilitate biomass granulation. 2) Granulation enhances the sludge settling property and retention time, which contributes to the extremely high nitrogen removal rate of 77 kg m−3 d−1 of Anammox upflow reactors. 3) Flotation of Anammox granules frequently occurs under nitrogen removal rate higher than 10 kg m−3 d−1, which is mainly due to the overproduction of EPS under high hydraulic conditions.

Rapid Formation of Aerobic Granular Sludge and Its Mechanism in a Continuous-Flow Bioreactor.

Based on the principle of self-coagulation of microorganisms, the flocculant-producing denitrifying bacterial TN-14 sludge was added to the continuous-flow reactor for treating domestic sewage. The bacterial TN-14 sludge acted as the main seed sludge to promote the rapid formation of aerobic granular sludge. The sludge morphology, sludge volume index (SVI) values, amounts of extracellular polymeric substances (EPS), and the role of calcium in the granulation process of the sludge were investigated. Results showed that brown aerobic granules with the particle size of 0.5 ~ 2.0mm was successfully cultivated at 40days, and its SVI30 decreased from 122.62mLg-1 initially to 46.61mLg-1 and remained at 44.28 ~ 60.51mLg-1 afterwards. The protein (PN) content in sludge EPS increased from 76.4mgg-1 initially to 512.3mgg-1. Compared with PN, the polysaccharide (PS) content did not change much throughout the operation process of the bioreactor. Energy-dispersive spectrum (EDS) showed that Ca elements were deposited inside the granular sludge, and X-ray diffraction (XRD) showed that Ca elements existed in the granular sludge in the forms of CaCO3, K2CaP2O7, Ca2P2O7, and Ca4O(PO4)2. The formation mechanism of continuous-flow aerobic granular sludge was that bacterial TN-14 sludge could promote the EPS content of sludge, and PN content of EPS increases the hydrophobicity and settling performance of the sludge. Calcium mainly exists in the granular sludge in the form of inorganic calcium phosphate, and therefore plays the role of nucleation in sludge granulation.

Effect of increase in salinity on ANAMMOX–UASB reactor stability

ABSTRACTThe effect of salinity on the anaerobic ammonium oxidation (ANAMMOX) process in a UASB reactor was investigated by analysing ammonium, nitrite, nitrate and TN concentrations, and TN removal efficiency. Extracellular polymeric substances (EPSs) and specific ANAMMOX activity (SAA) were evaluated. Results showed the effluent deteriorated after salinity was increased from 8 to 13 g/L and from 13 to 18 g/L, and TN removal efficiency decreased from 80% to 30% and 80% to 50%, respectively. However, ANAMMOX performance recovered and TN removal efficiency increased to 80% after 40 days when the influent concentrations of and were 200 mg/L and salinity levels were at 13 and 18 g/L, respectively. The amount of EPSs decreased from 58.9 to 37.1 mg/g volatile suspended solids (VSS) when the reactor was shocked by salinity of 13 g/L, and then increased to 57.2 mg/g VSS when the reactor recovered and ran stably at 13 g/L. The amount of EPSs decreased from 57.2 to 49.1 mg/g VSS when the reactor was shocked by salinity of 18 g/L, and then increased to 60.7 mg/g VSS when the reactor recovered and ran stably at 18 g/L. The amount of EPS and the amounts of polysaccharide, protein and humus showed no evident difference when the reactor recovered from different levels of salinity shocks. Batch tests showed salinity shock load from 8 to 38 g/L inhibited the SAA. However, when the reactor recovered from salinity shocks, SAA was higher compared to that when the reactor was subjected to the same level of salinity shock.

Harvesting of Scenedesmus obliquus using dynamic filtration with a perforated disk

Dynamic filtration adopting the rotation of a perforated disk was applied to harvest Scenedesmus obliquus. The highest rotation speed of the disk was found to give rise to 464 and 454 L/m2/h of plateau permeate flux for microfiltration (MF) membrane and ultrafiltration (UF) membrane, respectively, which were approximately 9 and 11 times higher than those with no rotation. This marked performance improvement for both filtration methods, with the UF exhibiting lower irreversible fouling, was attributed to the reduction of not only cake fouling (up to 97%) but also adsorption fouling (up to 71%) by way of high shear stress on the membrane surface. At the high rotation speed in which fouling was effectively suppressed, flux was linearly increased by trans-membrane pressure; at low speeds where fouling formation remained less disrupted, on the other hand, it reached a certain limit. Besides, fouling built and worsened particularly by high amounts of extracellular polymeric substance (EPS), which was as in the case of the seawater-supplemented culture, was effectively alleviated by fast rotation. The dynamic UF based on the rotation of the perforated disk was indeed a promising and suited means of harvesting S. obliquus, especially cells grown under economical yet EPS-inducing cultivation conditions.

Is biofilm removal properly assessed? Comparison of different quantification methods in a 96-well plate system.

Various methods have been reported to quantify total biofilm or different components of biofilm; however, these methods are often confusedly used, leading to discrepancies and misleading results. In this study, different methods for quantification of biofilm, including those for total biomass, total amount of bacterial cells, viable cell number, and amount of extracellular polymeric substances, were systematically compared in microtiter plates. To evaluate which method is suitable for assessment of biofilm removal and for bacterial killing, biofilm samples were treated with various cleaners possessing removing and/or killing capacities. It was found that most of the methods tested in this study in general exhibited high reproducibility and repeatability. Crystal Violet staining was a simple but reliable method for total biomass quantification. Total bacteria cell numbers could be reliably quantified by the fluorescent DNA-binding dye Acridine Orange. Viable cells could be quantified by either an ATP-based assay or a proliferation assay. Both of these viability methods showed a broad detection range and led to precise measurement. For quantification of proteins in the biofilm, staining with fluorescein isothiocyanate was most suitable. Furthermore, it was revealed that a combination of different methods is required to determine if a cleaner kills or removes biofilm.Electronic supplementary materialThe online version of this article (doi:10.1007/s00253-016-7396-9) contains supplementary material, which is available to authorized users.

A Comparative Study of the Biodesulfurization Efficiency of Acidithiobacillus ferrooxidans LY01 Cells Domesticated with Ferrous Iron and Pyrite

ABSTRACTThe high-sulfur coal desulfurization process completed by A. ferrooxidans LY01 cells domesticated with either ferrous iron [Fe(II)] or pyrite (FeS2) was investigated in this article. The desulfurization rate for 13 d was as high as 67.8% for the LY01 cells domesticated with pyrite but was only 45.6% for the LY01 cells domesticated with Fe(II). Bacterial adsorption experiments indicated that the bacterial adsorption quantity onto the pyrite particles was similar to the desulfurization efficiency. FTIR analysis showed that chemical composition of the two cell types was similar, but the LY01 cells domesticated with pyrite had higher levels of hydrophobic aromatic R-O groups than cells domesticated with Fe(II). The amount of extracellular polymeric substances (EPS) from the pyrite-domesticated LY01 cells was 1820 μg C/1010 cells, which was five times more than the amount of EPS in the Fe(II)-domesticated cells; the EPS readily bound Fe(III) with a maximum binding capacity of 0.21 mg Fe(III) per mg C EPS. Strains of pyrite-domesticated LY01 with a high amount of Fe(III) in their EPS possess greater oxidation activity than Fe(II)-domesticated strains with fewer Fe(III). These experiments showed the importance of the substrate-specific differences in the oxidative activity of A. ferrooxidans LY01. In addition, this study provides theoretical guidance for the future optimization of the biodesulfurization process.

Understanding of aerobic granulation enhanced by starvation in the perspective of quorum sensing.

Three sequencing batch reactors (M1, M2, and M3) were set up to investigate the influence of different lengths of starvation time (3, 5, and 7h) on aerobic granulation in the perspective of quorum sensing (QS). Autoinducer-2 (AI-2) level was quantified to evaluate the QS ability of aerobic granules. The results indicated that AI-2 level increased steadily during a cycle of sequencing batch reactors, suggesting that starvation was closely related to AI-2 secretion. In the long-term operation, aerobic granules cultivated using a prolonged starvation period had a better integrity and a higher level of cell adhesiveness despite a slower formation speed. With the extension of the starvation period, the total amount of extracellular polymeric substances (EPS) displayed an increasing tendency. EPS with large molecular weight (MW) also reached a higher level using a prolonged starvation period. However, a higher level of AI-2 and cell adhesiveness was observed in M2, which might be related to more stable granules. The results implied that the starvation period could trigger AI-2 secretion and promoted the production of large MW EPS, leading to cell adhesiveness enhancement and granule formation. Therefore, a combination of different starvation periods was proposed in this study in order to improve aerobic granulation.

Mucilaginibacter ginsengisoli sp. nov., isolated from a ginseng-cultivated soil.

A dark-pink-coloured bacterial strain, B4Y-8T, was isolated from a soil cultivated with ginseng. The 16S rRNA gene sequence of this strain showed highest similarity with Mucilaginibacter litoreus BR-18T (96.8 %), Mucilaginibacter lutimaris BR-3T (96.6 %) and Mucilaginibacter defluvii A5T (96.2 %) among the type strains of species of the genus Mucilaginibacter. Strain B4Y-8T was a strictly aerobic, Gram-stain-negative, non-motile, short-rod-shaped bacterium producing a large amount of extracellular polymeric substance. The strain grew at 10-35 °C (optimum, 25 °C), at pH 3.0-11.0 (optimum, pH 7.0) and in the presence of 0-1 % (w/v) NaCl (optimum, 0 %). The DNA G+C content of strain B4Y-8T was 49.0 mol%. It contained menaquinone 7 (MK-7) as the major isoprenoid quinone, and summed feature 3 (C16 : 1ω6c and/or C16 : 1ω7c) and iso-C15 : 0 as the major fatty acids. On the basis of evidence from the present polyphasic taxonomic study, strain B4Y-8T should be classified as representing a novel species of the genus Mucilaginibacter, for which the name Mucilaginibacter ginsengisoli sp. nov. is proposed. The type strain is B4Y-8T ( = KACC 18152T = JCM 30759T).

Biofouling in forward osmosis and reverse osmosis: Measurements and mechanisms

We have investigated biofouling behavior in forward osmosis (FO) and reverse osmosis (RO) membrane systems. Analysis of these two systems was done via comparison of biofilm structure and membrane permeate water flux decline. Experiments were performed using a model wastewater solution inoculated with Pseudomonas aeruginosa in laboratory-scale FO and RO test cells. For a meaningful comparison, biofouling runs were carried out using the same thin-film composite polyamide FO membrane, identical hydrodynamic conditions, and an initial permeate water flux of 19Lm−2h−1. Water flux decline after 600mL of cumulative permeate volume (or ~17–18h of fouling) was significantly lower in FO (~10%) compared to RO (~30%). FO biofilms grew in a loosely organized thick layer, with eminent mushroom-shaped structures, while RO biofilms grew in tightly organized mats encased in larger amounts of extracellular polymeric substances (EPS) per cell. The more compact biofilms in RO induced greater biofilm-enhanced osmotic pressure and hydraulic resistance to water flow compared to FO, which resulted in higher flux decline. We attribute the differences in biofouling behaviors in FO and RO to the different driving forces: osmotic pressure in FO and hydraulic pressure in RO.

Dual Roles of Capsular Extracellular Polymeric Substances in Photocatalytic Inactivation of Escherichia coli: Comparison of E. coli BW25113 and Isogenic Mutants.

The dual roles of capsular extracellular polymeric substances (EPS) in the photocatalytic inactivation of bacteria were demonstrated in a TiO2-UVA system, by comparing wild-type Escherichia coli strain BW25113 and isogenic mutants with upregulated and downregulated production of capsular EPS. In a partition system in which direct contact between bacterial cells and TiO2 particles was inhibited, an increase in the amount of EPS was associated with increased bacterial resistance to photocatalytic inactivation. In contrast, when bacterial cells were in direct contact with TiO2 particles, an increase in the amount of capsular EPS decreased cell viability during photocatalytic treatment. Taken together, these results suggest that although capsular EPS can protect bacterial cells by consuming photogenerated reactive species, it also facilitates photocatalytic inactivation of bacteria by promoting the adhesion of TiO2 particles to the cell surface. Fluorescence microscopy and scanning electron microscopy analyses further confirmed that high capsular EPS density led to more TiO2 particles attaching to cells and forming bacterium-TiO2 aggregates. Calculations of interaction energy, represented by extended Derjaguin-Landau-Verwey-Overbeek (XDLVO) potential, suggested that the presence of capsular EPS enhances the attachment of TiO2 particles to bacterial cells via acid-base interactions. Consideration of these mechanisms is critical for understanding bacterium-nanoparticle interactions and the photocatalytic inactivation of bacteria.

Removal of trace metals and improvement of dredged sediment dewaterability by bioleaching combined with Fenton-like reaction

Bioleaching by Aspergillus niger strain SY1 combined with Fenton-like reaction was optimized to improve trace metal removal and dewaterability of dredged sediments. The major optimized parameters were the duration of bioleaching and H2O2 dose in Fenton-like process (5 days and 2g H2O2/L, respectively). Bioleaching resulted in the removal of ≈90% of Cd, ≈60% of Zn and Cu, ≈20% of Pb, and in decrease of sediment pH from 6.6 to 2.5 due to organic acids produced by A. niger. After addition of H2O2, Fenton-like reaction was initiated and further metal removal occurred. Overall efficiency of the combined process comprised: (i) reduction of Cd content in sediment by 99.5%, Cu and Zn by >70% and Pb by 39% as a result of metal release bound in all mobilizable fractions; (ii) decrease of sediment capillary suction time (CST) from 98.2s to 10.1s (by 89.8%) and specific resistance to filtration (SRF) from 37.4×1012m/kg to 6.2×1012m/kg (by 83.8%), due to reducing amount of extracellular polymeric substances (EPS) by 68.7% and bound water content by 79.1%. The combined process was found to be an efficient method to remove trace metals and improve dewaterability of contaminated dredged sediments.

Temperature and Concentration Effects on Biodegradation Kinetics and Surface Properties of Phenol Degrading Rhodotorula glutinis

Biotoxicity and low temperature are among the challenges of biodegradation as well as microbial transport in in-situ bioremediation applications. This study investigates the effect of low temperature, high and low concentrations of phenol, on biodegradability and the surface properties of Rhodotorula glutinis which may be instrumental to develop control mechanisms in engineering applications. Results showed that Rhodotorula glutinis is able to grow at low phenol (< 0.5 g/L) and glucose (1.0 g/L) concentrations. Microbial growth rates of Rhodotorula glutinis is the highest at 25°C for phenol fed reactors. Temperature and type of carbon source affected the total amount of extracellular polymeric substances (EPS) and the relative amounts of EPS constituents. Phenol fed Rhodotorula glutinis, at 4°C, were found to be hydrophobic whereas Rhodotorula glutinis showed less hydrophobic characteristics at 25°C, regardless of the carbon source. Surface hydrophobicity of Rhodotorula glutinis is related to the distribution of carbohydrate and proteins in the EPS.

Possible causes of excess sludge reduction adding metabolic uncoupler, 3,3′,4′,5-tetrachlorosalicylanilide (TCS), in sequence batch reactors

Two parallel sequence batch reactors (SBRs) were operated, with and without TCS addition, to research the causes of sludge reduction by uncouplers. Three possible mechanisms of sludge reduction by TCS were studied: (1) occurrence of metabolic uncoupling, (2) consumption of more energy to resist the infection of TCS, (3) promotion of lysis–cryptic growth by TCS addition. Results showed the remarkable reduction of electronic transport system (ETS) activity and specific cellular ATP (SATP) in TCS reactor, which proved the occurrence of metabolic uncoupling. The increasing amounts of extracellular polymeric substances (EPS), as measured by chemical methods and excitation–emission matrix (EEM) fluorescence spectra, implied microorganisms consumed more energy to resist TCS. The similar DNA concentrations of the effluents in two reactors indicated sludge lysis was not intensified by TCS. Therefore, uncoupler might not only cause metabolic uncoupling but also induce more energy consumption in the production of some substances to resist uncoupler.

Biofouling of reverse-osmosis membranes under different shear rates during tertiary wastewater desalination: Microbial community composition

We investigated the influence of feed-water shear rate during reverse-osmosis (RO) desalination on biofouling with respect to microbial community composition developed on the membrane surface. The RO membrane biofilm's microbial community profile was elucidated during desalination of tertiary wastewater effluent in a flat-sheet lab-scale system operated under high (555.6 s−1), medium (370.4 s−1), or low (185.2 s−1) shear rates, corresponding to average velocities of 27.8, 18.5, and 9.3 cm s−1, respectively. Bacterial diversity was highest when medium shear was applied (Shannon–Weaver diversity index H' = 4.30 ± 0.04) compared to RO-membrane biofilm developed under lower and higher shear rates (H′ = 3.80 ± 0.26 and H′ = 3.42 ± 0.38, respectively). At the medium shear rate, RO-membrane biofilms were dominated by Betaproteobacteria, whereas under lower and higher shear rates, the biofilms were dominated by Alpha- and Gamma- Proteobacteria, and the latter biofilms also contained Deltaproteobacteria. Bacterial abundance on the RO membrane was higher at low and medium shear rates compared to the high shear rate: 8.97 × 108 ± 1.03 × 103, 4.70 × 108 ± 1.70 × 103 and 5.72 × 106 ± 2.09 × 103 copy number per cm2, respectively. Interestingly, at the high shear rate, the RO-membrane biofilm's bacterial community consisted mainly of populations known to excrete high amounts of extracellular polymeric substances. Our results suggest that the RO-membrane biofilm's community composition, structure and abundance differ in accordance with applied shear rate. These results shed new light on the biofouling phenomenon and are important for further development of antibiofouling strategies for RO membranes.