Presence Of Extracellular Polymeric Substances Research Articles

Pseudomonas aeruginosa infection in cystic fibrosis patients

Cystic Fibrosis is an autosomal recessive genetic disease where the lung is heavily colonized by biofilm forming Pseudomonas aeruginosabacteria. Biofilm due to its thick wall and presence of extracellular polymeric substances confers bacteria with a much higher resistance against antimicrobial compounds and environmental stress. Research has revealed the presence of extracellular DNA (eDNA) in significant amounts in the biofilm walls of such bacteria. Although the exact role of eDNA has not been elucidated but research prompts that eDNA to have significant role in stabilizing the biofilm complex structure as well as providing the bacteria with resistance against antimicrobial and antibiotic compounds. The presence of eDNA prompted research to use DNase as a biofilm destabilizing agent. Under experimental conditions, DNase has shown to dissolve early forming biofilm walls especially when used in combination with antibiotics therefore suggesting the use of DNase as an effective treatment strategy in CF patients. But ambiguity arises when some recent studies showed that P. aeruginosa can opportunistically use DNase for its survival advantage by using the enzyme to access phosphate, carbon and nitrogen constituentsof the DNA in nutrient limited conditions; and also by using the DNase to degrade the DNA component of neutrophil extracellular traps. These apparent contradictions call for more thorough investigations into the role of eDNA and DNase in CF biofilm infection. DOI: http://dx.doi.org/10.3329/jdmc.v22i1.15689 J Dhaka Medical College, Vol. 22, No. 1, April, 2013, Page 77-86

Phosphatase activity and specific methanogenic activity in an anaerobic reactor treating sludge from a brackish recirculation aquaculture system

Anaerobic treatment of high salinity sludge from marine/brackish recirculation aquaculture systems is potentially limited by inhibition of enzymatic activities and cell lysis resulting from high osmotic pressures. To further address these limitations the following investigations were conducted: effect of salinity on phosphatase activity (PA), soluble microbial products (SMP) production, and presence of extracellular polymeric substances (EPS); effect of iron (III) chloride (FeCl3) on PA and specific methanogenic activity (SMA); effect of addition of the compatible solute glycine betaine (GB) and potassium on PA, as well as on SMP and EPS production, all under saline conditions. The results show that salinity has different effects on PA of anaerobes under starvation and feeding conditions. FeCl3 increased the SMA of the sludge by 22.5% at 100 mg FeCl3/L compared with a control group (0 mg FeCl3/L). Furthermore, results of analysis of variance tests show that betaine increased the polysaccharide content of EPS and polypeptide content of SMP. However, addition of 1 mM potassium chloride did not show a significant effect on EPS and SMP composition. In conclusion, anaerobic digestion of salty sludges from a brackish aquaculture recirculation system may not be negatively affected by FeCl3 addition to concentrate waste streams, whereas GB boosts the production of SMP and EPS.

Investigating why recycling gravity harvested algae increases harvestability and productivity in high rate algal ponds

It has previously been shown that recycling gravity harvested algae promotes Pediastrum boryanum dominance and improves harvestability and biomass production in pilot-scale High Rate Algal Ponds (HRAPs) treating domestic wastewater. In order to confirm the reproducibility of these findings and investigate the mechanisms responsible, this study utilized twelve 20 L outdoor HRAP mesocosms operated with and without algal recycling. It then compared the recycling of separated solid and liquid components of the harvested biomass against un-separated biomass. The work confirmed that algal recycling promoted P. boryanum dominance, improved 1 h-settleability by >20% and increased biomass productivity by >25% compared with controls that had no recycling. With regard to the improved harvestability, of particular interest was that recycling the liquid fraction alone caused a similar improvement in settleability as recycling the solid fraction. This may be due to the presence of extracellular polymeric substances in the liquid fraction. While there are many possible mechanisms that could account for the increased productivity with algal recycling, all but two were systematically eliminated: (i) the mean cell residence time was extended thereby increasing the algal concentration and more fully utilizing the incident sunlight and, (ii) the relative proportions of algal growth stages (which have different specific growth rates) was changed, resulting in a net increase in the overall growth rate of the culture.

Removal characteristics of engineered nanoparticles by activated sludge

Environmental release of engineered nanoparticles (NPs) has been on the rise due to the increased use of NPs in commercial products. In addition, the fate of NPs in sewage treatment processes may play an important role in determining the environmental release pathway of NPs. In this study, we investigated the removal of engineered NPs (AgNPs, TiO2NPs, and SiO2NPs) using activated sludge by evaluating the effects of several important factors of the NPs, including physicochemical properties, contact time between NPs and activated sludge, aquatic chemistry of sewage, and the presence of extracellular polymeric substances (EPS) in the activated sludge. For all three types of NPs tested, a considerable amount of NPs were removed after exposure to activated sludge in a time-dependent manner; nevertheless, the removal efficiencies depended on the type of NPs and seemed to be affected by the NP stability relative to the hydrodynamic diameter (HDD) and zeta potential. In addition, the presences of both ionic compounds and EPS significantly enhanced the NP removal efficiency, indicating that the instability of the NPs resulting from the ionic strength in sewage and entrapment of NP by EPS played an important role in NP removal by activated sludge. These results suggest that the removal efficiencies can be affected by the operating conditions of the activated sludge process and the conditions of the activated sludge; therefore, these factors should be considered when developing approaches to sufficiently remove NPs from sewage treatment plants.

Citrate-Capped Silver Nanoparticles Showing Good Bactericidal Effect against Both Planktonic and Sessile Bacteria and a Low Cytotoxicity to Osteoblastic Cells

A common problem with implants is that bacteria can form biofilms on their surfaces, which can lead to infection and, eventually, to implant rejection. An interesting strategy to inhibit bacterial colonization is the immobilization of silver (Ag) species on the surface of the devices. The aim of this paper is to investigate the action of citrate-capped silver nanoparticles (AgNPs) on clinically relevant Gram-positive (Staphylococcus aureus) and Gram-negative (Pseudomonas aeruginosa) bacteria in two different situations: (i) dispersed AgNPs (to assess the effect of AgNPs against planktonic bacteria) and (ii) adsorbed AgNPs on titanium (Ti) substrates, a material widely used for implants (to test their effect against sessile bacteria). In both cases, the number of surviving cells was quantified. The small amount of Ag on the surface of Ti has an antimicrobial effect similar to that of pure Ag surfaces. We have also investigated the capability of AgNPs to kill planktonic bacteria and their cytotoxic effect on UMR-106 osteoblastic cells. The minimum bactericidal concentration found for both strains is much lower than the AgNP concentration that leads to cytotoxicity to osteoblasts. Planktonic P. aeruginosa show a higher susceptibility to Ag than S. aureus, which can be caused by the different wall structures, while for sessile bacteria, similar results are obtained for both strains. This can be explained by the presence of extracellular polymeric substances in the early stages of P. aeruginosa biofilm formation. Our findings can be important to improving the performance of Ti-based implants because a good bactericidal action is obtained with very small quantities of Ag, which are not detrimental to the cells involved in the osseointegration process.

Enhanced resistance to nanoparticle toxicity is conferred by overproduction of extracellular polymeric substances

The increasing production and use of engineered nanoparticles, coupled with their demonstrated toxicity to different organisms, demands the development of a systematic understanding of how nanoparticle toxicity depends on important environmental parameters as well as surface properties of both cells and nanomaterials. We demonstrate that production of the extracellular polymeric substance (EPS), colanic acid by engineered Escherichia coli protects the bacteria against silver nanoparticle toxicity. Moreover, exogenous addition of EPS to a control strain results in an increase in cell viability, as does the addition of commercial EPS polymer analogue xanthan. Furthermore, we have found that an EPS producing strain of Sinorhizobium meliloti shows higher survival upon exposure to silver nanoparticles than the parent strain. Transmission electron microscopy (TEM) observations showed that EPS traps the nanoparticles outside the cells and reduces the exposed surface area of cells to incoming nanoparticles by inducing cell aggregation. Nanoparticle size characterization in the presence of EPS and xanthan indicated a marked tendency towards aggregation. Both are likely effective mechanisms for reducing nanoparticle toxicity in the natural environment.

Biofilm Formation in Milk Production and Processing Environments; Influence on Milk Quality and Safety

Abstract: Bacteria in milk have the ability to adhere and aggregate on stainless steel surfaces, resulting in biofilm formation in milk storage tanks and milk process lines. Growth of biofilms in milk processing environments leads to increased opportunity for microbial contamination of the processed dairy products. These biofilms may contain spoilage and pathogenic microorganisms. Bacteria within biofilms are protected from sanitizers due to multispecies cooperation and the presence of extracellular polymeric substances, by which their survival and subsequent contamination of processed milk products is promoted. This paper reviews the most critical factors in biofilm formation, with special attention to pseudomonads, the predominant spoilage bacteria originating from raw milk. Biofilm interactions between pseudomonads and milk pathogens are also addressed, as emerging risks and future research perspectives, specifically related to the milk processing environment.

Biofilm formation for organic matter and sulphate removal in gas-lift reactors

A start-up strategy was presented and evaluated to obtain a well-established biofilm in a gas lift-reactor capable both for the removal of organic matter and sulphate. Pumice stone was used as material support. The influence of shear forces, given by the biogas recirculation, the effect of the COD/SO 4 -2 ratio and the OLRs increase were evaluated on the reactor performance. From the first stages, cell colonization was observed along with the presence of extracellular polymeric substances. The COD and sulphate removal was over 70%, for all conditions. The increase of gas flow did not have an adverse effect on biofilm development even though there was some detachment. Specific methanogenic activity of the biofilm increased along the experiments. Operational parameters as alkalinity and alkalinity ratio were within the recommended values for the operation with sulphate-rich wastewater. For gas-lift reactors operation it becomes fundamental to have a suitable start-up strategy that takes into account the initial biofilm development from a non-acclimatized biomass.

Effect of EPS on adhesion of Acidithiobacillus ferrooxidans on chalcopyrite and pyrite mineral surfaces

Extracellular polymeric substances (EPS) were extracted from Acidithiobacillus ferrooxidans through sonication method associated with centrifugation, which was evaluated tentatively with 2-keto-3-deoxyoctonic acid (KDO) as the indicator of EPS by spectrophotometry. Then the effect of EPS of A. ferrooxidans on the adhesion on chalcopyrite and pyrite surfaces was studied through a series of comparative experiments. The untreated cells and EPS-free cells of A. ferrooxidans were mixed with EPS suspension, Fe 2+ or Fe 3+, respectively. The planktonic cells were monitored in 2 h during bioleaching. The results indicate that the presence of EPS on the cell is an important factor for the adhesion to chalcopyrite and pyrite. A decrease of attachment of A. ferrooxidans to minerals was produced by the deficiency of EPS, which can recover mostly when the EPS was re-added into the EPS-free cells. The restoring extent is more obvious in pyrite than in chalcopyrite. The extent of cell adhesion to chalcopyrite increased when EPS and Fe 3+ added, and decreased when Fe 2+ added, which imply the electrostatic interaction plays a main role in initial adhesion between bacteria and minerals and it is a driving force for bacteria to produce EPS probably as a result of regaining their attachment ability to copper sulphides.

Role of extracellular polymeric substances in Cu(II) adsorption on Bacillus subtilis and Pseudomonas putida

The effect of extracellular polymeric substances (EPS) of Gram-positive Bacillus subtilis and Gram-negative Pseudomonas putida on Cu(II) adsorption was investigated using a combination of batch adsorption, potentiometric titrations, Fourier transform infrared spectroscopy. Both the potentiometric titrations and the Cu(II) adsorption experiments indicated that the presence of EPS in a biomass sample significantly enhance Cu(II) adsorption capacity. Surface complexation modeling showed that the pKa values for the three functional groups (carboxyl, phosphate and hydroxyl) were very similar for untreated and EPS-free cells, indicating no qualitative difference in composition. However, site concentrations on the untreated cell surface were found to be significantly higher than those on the EPS-free cell surface. Infrared analysis provided supporting evidence and demonstrated that carboxyl and phosphate groups are responsible for Cu(II) adsorption on the native and EPS-free cells.

Microscopical Observation of the Marine Bacterium Vibrio Natriegeus Growth on Metallic Corrosion

Biofilm formation and the adsorption of nonsulfate-reducing marine bacterium Vibrio natriegens (V. natriegens) to different metallic surfaces have been studied by atomic force microscopy (AFM). The nature of extracellular polymeric substance (EPS) has been monitored through the force-distance curve. EPS may act as adhesive binding adjacent cells and thus further enhance the aggregation of microbes during cluster formation due to its adhesiveness in nature. The AFM images supplied the change of the morphology of microbial cells and colonies, the distribution of microbial colonies, and the presence of EPS on the surfaces with immersion. Results showed that the biofilms formed on the surfaces increased in thickness and heterogeneity with time and that 3–7 days was the proliferation period of the bacteria on metallic surfaces. The corrosion type of the metals by V. natriegens could be confirmed as typical pitting attack through scanning electron microscopy (SEM) observation.

Theory of Microbial Carbonate Precipitation and Its Application in Restoration of Cement‐based Materials Defects

AbstractBacterial induced carbonate mineralization has been demonstrated as a new potential method for restoration of limestones in historic buildings and monuments. We claim here the formation of calcium carbonate was controlled by extracellular polymeric substances (EPS) isolated from Bacillus pasteurii. The process of crystallization nucleation was accelerated in the presence of cells and inhibited in the presence of EPS. The CaCO3 film deposited on cement paste surface was about 100 µm after 7 d treatment. The results of various restoring methods showed that higher decrease of water absorption of cement paste was gained in brushing application in the presence of agar, which could maintain urease with high activity in long term compared to spraying method. The coefficient of capillary suction of cement paste treated with brushing method was reduced by 90%. Mixed media consisted of sands, urea, Ca2+ and concentrated biomass, was injected into artificial cracks of cement paste followed by continual nutrient supplement, and CaCO3 particles were precipitated gradually between sands particles which were combined with cement matrix. The results showed that the compressive strength of recovered specimens was restored to 84%, which demonstrated that this kind of bio‐restoration method is effective in repairing surface defects of cement‐based materials.

Contribution of Extracellular Polymeric Substances on Representative Gram Negative and Gram Positive Bacterial Deposition in Porous Media

The significance of extracellular polymeric substances (EPS) on cell transport and retained bacteria profiles in packed porous media (quartz sand) was examined by direct comparison of the overall deposition kinetics and retained profiles of untreated bacteria (with EPS) versus those of treated cells (without EPS) from the same cell type. Four representative cell types, Pseudomonas sp. QG6 (gram-negative, motile), mutant Escherichia coli BL21 (gram-negative, nonmotile), Bacillus subtilis (gram-positive, motile), and Rhodococcus sp. QL2 (gram-positive, nonmotile), were employed to systematically determine the influence of EPS on cell transport and deposition behavior. Packed column experiments were conducted for the untreated and treated cells in both NaCl (four ionic strength ranging from 2.5 mM to 20 mM) and CaCl(2) (5 mM) solutions at pH 6.0. The breakthrough plateaus of untreated bacteria were lower than those of treated bacteria for all four cell types under all examined conditions (in both NaCl and CaCl(2) solutions), indicating that the presence of EPS on cell surfaces enhanced cell deposition in porous media regardless of cell type and motility. Retained profiles of both untreated and treated cells for all four cell types deviated from classic filtration theory (log-linear decreases). However, the degree of deviation was greater for all four untreated cells, indicating that the presence of EPS on cell surfaces increased the deviation of retained profiles from classic filtration theory. Elution experiments demonstrated that neither untreated nor treated cells preferentially deposited in secondary energy minima. Furthermore, the release of previously deposited cells in the secondary energy minima did not change the shape of retained cell profiles, indicating that deposition in secondary energy minima did not produce the observed deviations of retained profiles from classic filtration theory.

Structure and Composition of Aggregates in Two Large European Rivers, Based on Confocal Laser Scanning Microscopy and Image and Statistical Analyses

Floating riverine aggregates are composed of a complex mixture of inorganic and organic components from their respective aquatic habitats. Their architecture and integrity are supplemented by the presence of extracellular polymeric substances of microbial origin. They are also a habitat for virus-like particles, bacteria, archaea, fungi, algae, and protozoa. In this study we present different confocal laser scanning microscopy strategies to examine aggregates collected from the Danube and Elbe Rivers. In order to collect multiple types of information, various approaches were necessary. Small aggregates were examined directly. To analyze large and dense aggregates, limitations of the technique were overcome by cryo-sectioning and poststaining of the samples. The staining procedure included positive staining (specific glycoconjugates and cellular nucleic acid signals) as well as negative staining (aggregate volume) and multichannel recording. Data sets of cellular nucleic acid signals (CNAS) and the structure of aggregates were visualized and quantified using digital image analysis. The Danube and Elbe Rivers differed in their aggregate composition and in the relative contribution of specific glycoconjugate and CNAS volume to the aggregate volume; these contributions also changed over time. We report different spatial patterns of CNAS inside riverine aggregates, depending on aggregate size and season. The spatial structure of CNAS inside riverine aggregates was more complex in the Elbe River than in the Danube River. Based on our samples, we discuss the strengths and challenges involved in scanning and quantifying riverine aggregates.

A vibrating membrane bioreactor (VMBR): Macromolecular transmission—influence of extracellular polymeric substances

The vibrating membrane bioreactor (VMBR) system facilitates the possibility of conducting a separation of macromolecules (BSA) from larger biological components (yeast cells) with a relatively high and stable macromolecular transmission at sub-critical flux. This is not possible to achieve for a static non-vibrating membrane module. A BSA transmission of 74% has been measured in the separation of 4 g/L BSA from 8 g/L dry weight yeast cells in suspension at sub-critical flux ( 20 L / ( m 2 h ) ) . However, this transmission is lower than the 85% BSA transmission measured for at pure 4 g/L BSA solution. This can be ascribed to the presence of extracellular polymeric substances (EPS) from the yeast cells. The initial fouling rate for constant sub-critical flux filtration of unwashed yeast cells is 3–4 times larger than for washed yeast cells ( 18 ( mbar / h ) / 5 ( mbar / h ) ) . At sub-critical flux, an EPS transmission of around 32% is measured for a pure yeast cell suspension. Thus, EPS and BSA are “competing” in being transmitted which might explain the lowered BSA transmission in the presence of yeast cells. Additionally, EPS heavily foul the membranes, leading to a 86% permeability drop and a fouling resistance 6 times larger than the membrane resistance after 5 1 2 h of constant sub-critical flux filtration ( 20 L / ( m 2 h ) ) of pure 8 g/L dry weight yeast cell suspensions. Thus, the addition of hydraulic resistance caused by EPS might also explain the lowered BSA transmission, in the presence of yeast cells, since the membrane pores might be narrowed or partly blocked. EPS is, furthermore, able to cause a relatively large permeability drop even on a membrane module pre-fouled by EPS.

The Influence of the Marine Bacterium <i>Vibrio Natriegens</i> Growth on Metallic Corrosion as Studied by Microscope Technologies

Biofilm formation and the adsorption of nonsulfate-reducing marine bacterium Vibrio natriegens (V. natriegen) to different metallic surfaces have been studied by atomic force microscopy (AFM). The nature of extracellular polymeric substance (EPS) has been monitored through the force-distance curve. EPS may act as adhesive binding adjacent cells and thus further enhance the aggregation of microbes during cluster formation due to its adhesiveness in nature. The AFM images supply the change of the morphology of microbial cells and colonies, the distribution of microbial colonies and the presence of EPS on the surfaces with immersion. Results show that the biofilms formed on the surfaces increase in thickness and heterogeneity with time and that 3~7 days is the proliferation period of the bacteria on metallic surfaces. The corrosion type of the metals by V. natriegens could be confirmed as typical pitting attack through scanning electron microscopy (SEM) observation.

Correlation betweenEnterococcus faecalisBiofilms Development Stage and Quantitative Surface Roughness Using Atomic Force Microscopy

Biofilms are assemblages of microorganisms and their associated extracellular products at an interface and typically with an abiotic or biotic surface. The study of the morphology of biofilms is important because they are associated with processes of biofouling, corrosion, catalysis, pollutant transformation, dental caries, drug resistance, and so forth. In the literature, biofilms have been examined by atomic force microscopy (AFM), which has proven to be a potent tool to study different aspects of the biofilm development on solid surfaces. In this work, we used AFM to investigate topographical changes during the development process of Enterococcus faecalis biofilms, which were generated on sterile cellulose nitrate membrane (CNM) filters in brain heart infusion (BHI) broth agar blood plates after 24, 36, 72, 192, and 360 h. AFM height images showed topographical changes due to biofilm development, which were used to characterize several aspects of the bacterial surface, such as the presence of extracellular polymeric substance, and the biofilm development stage. Changes in the development stage of the biofilm were shown to correlate with changes in the surface roughness as quantified through the mean roughness.

Influence of Extracellular Polymeric Substances onPseudomonas aeruginosaTransport and Deposition Profiles in Porous Media

The impact of cell surface extracellular polymeric substances (EPS) on bacterial transport and retention profiles was investigated in saturated columns packed with glass beads. Three genetically well-defined isogenic Pseudomonas aeruginosa strains with different EPS secretion capability and EPS composition were used to systematically examine their deposition behavior over a range of solution chemistry. The presence of EPS on nonmucoid strain PA01 and mucoid strain PD0300 significantly increased bacterial adhesion over the EPS deficient PA01 psl pel mutant strain despite their similar surface charge as indicated by the zeta potential measurements. Retained bacterial profiles show the deposition rate coefficients with various shapes and degrees of deviation from those expected from the classic filtration theory. Non-monotonic deviations from the log-linear deposition pattern with the majority of the bacteria retained downgradient of the column inlet were observed when bacterial cells were encapsuled by EPS under both high and low ionic strength conditions. In contrast, the EPS-deficient strain exhibited monotonic deviation from theory only under low ionic strength conditions. The results demonstrate that the non-monotonic deviation from filtration theory observed in this study was driven by steric interactions between extracellular polymers and glass beads. Analysis of the retained polysaccharides (carbohydrates and uronic acids) and protein profiles suggests that bacterial re-entrainment and re-entrapment may have contributed to the downgradient movement of the maximum retained bacteria. The detachment of bacteria may leave behind various constituents of EPS as their "footprints," which can be a valuable tool for tracking the trajectory of bacterial transport.

Closing the water loop in a maltery: reuse tests at pilot-scale

This paper reports on the potential for water reuse in the malting sector. Core unit of a treatment train to close the water loop was a membrane bioreactor (MBR). We compared three different commercial submerged membranes for their fouling potential in this application and related this to the presence of extracellular polymeric substances (EPS). In a second step, we subjected MBR permeate to reverse osmosis and several (advanced) oxidation processes to evaluate the water quality achieved. Finally we performed a set of water reuse tests with waters obtained through different scenarios. The optimal scenario was then tested in a closed water loop over several malting cycles at pilot scale and the effect on water and malt quality was investigated.

The Role of Bioflocculation on Suspended Solids and Particulate COD Removal in the Trickling Filter Process

Understanding that there is a significant presence of extracellular polymeric substances at the biofilm/wastewater interface and that the primary constituent of chemical oxygen demand (COD) in domestic wastewaters is organic particulates, this research describes the kinetics of particulate removal in a pilot-scale trickling filter (TF) and the role of bioflocculation in the removal process. Recent research has described the role of bioflocculation on particulate COD (PCOD) removal in suspended growth biological wastewater treatment systems. However, no research pertaining to PCOD removal by bioflocculation in attached growth systems was identified prior to this study. For this study, experiments were conducted using both bench- and pilot-scale biofilm reactors and provided evidence that the removal of organic and inorganic particulate matter in a TF bioreactor follows a first-order bioflocculation rate equation. The statistical analysis of data obtained from the pilot TF fits the dispersion model to suspended solids and PCOD remaining in the pilot TF.