Extracellular Polymeric Substances Layer Research Articles

Impact of sludge cation distribution pattern on its filterability in membrane bioreactor

The distributions of cations of various valences (Na+, Ca2+ and Fe3+) in the outer layers of extracellular polymeric substances (EPSs) to pellet have a significant impact on the stratification structure of polysaccharides (PS) or proteins (PN) in activated sludge. Comparison with the control showed that the monovalent Na+ reduced flocculability slightly (about 9.75%), whereas Ca2+ and Fe3+ increased flocculability significantly. The modified fouling index (MFI) had a significant correlation with PN in the supernatant (rp=0.8593), slime (rp=0.7218) and loosely bound EPS (LB, rp=0.8012). However, it had a moderate correlation with PS in supernatant (rp=0.5842), and weak correlation to slime (rp=0.3785) or LB (rp=0.3219). There was an ignored correlation with PN or PS in the tightly bound EPS (TB) or pellet. The lower amount of PN or PS in the supernatant would have positive impact on improving the activated sludge filterability.

Isolation, characterization and localization of extracellular polymeric substances from the cyanobacterium Arthrospira platensis strain MMG-9

Arthrospira platensis is a cyanobacterium known for its nutritional value and secondary metabolites. Extracellular polymeric substances (EPS) are an important trait of most cyanobacteria, including A. platensis. Here, we extracted and analysed different fractions of EPS from a locally isolated strain of A. platensis. Three different fractions of EPS were distinguished. These were EPS released into the medium (REPS), EPS loosely bound to the organism (LEPS) and EPS tightly bound to the organism (TEPS), which were extracted by different procedures. The LEPS fraction was smaller than the other two fractions. The EPS of A. platensis exhibited high diversity. Total protein and carbohydrate content was determined in each of these fractions. The largest amount of total carbohydrates and total proteins was in the TEPS fraction. Eight sugar moieties were detected and analysed in all EPS fractions using HPAE-PAD. Fructose, mannose and ribose were rare sugar residues in all fractions of EPS. With the exception of fructose, all sugars tested for were detected in TEPS. The amount of sugars detected was significantly higher in TEPS compared with the two other fractions, especially for galactose, xylose and glucose. The EPS were localized by confocal laser scanning microscopy (CLSM) after staining with different fluorescent dyes and it was found that A. platensis possessed a thick and smooth layer of EPS around the spiral trichomes.

Effect of Exogenous Galactose on EPS Production during Bioleaching of Pyrite by <i>Acidithiobacillus ferrooxidans</i>

Extracellular polymeric substances (EPS) play an important role in the attachment of bacteria to sulphide minerals, biofilm formation and efficiency of the bioleaching process. Previous studies have suggested a potential connection between galactose and EPS formation. In this context, the influence of exogenous galactose on EPS formation during the bioleaching of pyrite was studied. In order to fully adapt the microorganism to bioleaching conditions it was performed a total of five consecutive sub cultures, one every fifteen days, taking for each one inocula from previous culture in shake flasks with 200 ml of fermentation medium at 30°C, 200 rpm, 40 gL-1 mineral and an initial pH of 1,8. Assays were performed in a medium supplemented with exogenous galactose (0.25% w/v) and without exogenous galactose (control), both with an initial concentration of ferric sulphate in the first three sub cultures of 5 gL-1, decreasing in the last two sub cultures to 2.5 gL-1. Samples of three cultures in both conditions were analyzed using confocal laser scanning microscopy (CLSM) labelling the cells with propidium iodide and EPS carbohydrates with Wheat Germ Agglutinin (WGA). Samples obtained on the last day of the fifth culture showed that the EPS layer on the particle surface was 5.00 μm3/μm2 in the case of the control condition and 6.10 μm3/μm2 when bioleaching was carried out in the presence of exogenous galactose. Also it was observed that in the fifth sub culture the volumetric productivity of total iron in the control experiment was 0.0065 gL-1.h-1 compared with 0.0076 gL-1.h-1 obtained in presence of galactose. The results reveal that the presence of galactose in the bioleaching solution stimulates EPS's formation and apparently also favour the pyrite bioleaching process.

Probing the fungicidal property of CdS quantum dots on Saccharomyces cerevisiae and Candida utilis using MALDI-MS

For the first time, we report the successful application of inhouse synthesized CdS quantum dots (QDs) with particle sizes between 1 and 7 nm exhibiting excellent fungicidal activity based on the interactions with Saccharomyces cerevisiae and Candida utilis. The growth curves and the growth rates of both fungi were established in the presence of three varying concentrations of CdS QDs. It was observed that the CdS QDs were highly inhibitory even at the lowest concentration of 10 mg/L used in this study, while the untreated control cells followed a normal growth pattern in the cases of both Saccharomyces and Candida. MALDI-MS was applied to substantiate the observations obtained by direct cell count method. It was observed that the trend observed in the case of Saccharomyces and Candida was well-represented in the MALDI-MS spectra. This study proposes a mechanism for the first time based on MALDI-MS results, that the CdS QDs interact with the extracellular polymeric substances (EPS) and remove small molecules from EPS layer; on the other hand, it was observed that CdS QDs at all concentrations lead to enrichment of protein signals in MALDI-MS. We have substantiated these results by quantifying the EPS in the control and treated cells and also using TEM to further confirm the results.

The surface properties of Shewanella putrefaciens 200 and S. oneidensis MR-1: the effect of pH and terminal electron acceptors.

BackgroundWe investigated the surface characteristics of two strains of Shewanella sp., S. oneidensis MR-1 and S. putrefaciens 200, that were grown under aerobic conditions as well as under anaerobic conditions with trimethylamine oxide (TMAO) as the electron acceptor. The investigation focused on the experimental determination of electrophoretic mobility (EPM) under a range of pH and ionic strength, as well as by subsequent modeling in which Shewanella cells were considered to be soft particles with water- and ion-permeable outermost layers.ResultsThe soft layer of p200 is significantly more highly charged (i.e., more negative) than that of MR-1. The effect of electron acceptor on the soft particle characteristics of Shewanella sp. is complex. The fixed charge density, which is a measure of the deionized and deprotonated functional groups in the soft layer polymers, is slightly greater (i.e., more negative) for aerobically grown p200 than for p200 grown with TMAO. On the other hand, the fixed charge density of aerobically grown MR1 is slightly less than that of p200 grown with TMAO. The effect of pH on the soft particle characteristics is also complex, and does not exhibit a clear pH-dependent trend.ConclusionsThe Shewanella surface characteristics were attributed to the nature of the outermost soft layer, the extracellular polymeric substances (EPS) in case of p200 and lypopolysaccharides (LPS) in case of MR1 which generally lacks EPS. The growth conditions (i.e., aerobic vs. anaerobic TMAO) have an influence on the soft layer characteristics of Shewanella sp. cells. Meanwhile, the clear pH dependency of the mechanical and morphological characteristics of EPS and LPS layers, observed in previous studies through atomic force microscopy, adhesion tests and spectroscopies, cannot be corroborated by the electrohydrodynamics-based soft particle characteristics which does not exhibited a clear pH dependency in this study. While the electrohydrodynamics-based soft-particle model is a useful tool in understanding bacteria’s surface properties, it needs to be supplemented with other characterization methods and models (e.g., chemical and micromechanical) in order to comprehensively address all of the surface-related characteristics important in environmental and other aqueous processes.

Engineered biosealant strains producing inorganic and organic biopolymers

Microbiologically induced calcium carbonate precipitation (MICCP) is a naturally occurring biological process that has shown its potential in remediation of a wide range of structural damages including concrete cracks. In this study, genetically engineered microorganisms, capable of producing extracellular polymeric substances (EPSs) as well as inducing MICCP, were developed based on the assumption that the complex of inorganic CaCO3 and organic EPS would provide a stronger matrix than MICCP alone as biosealant. In order to develop a recombinant biosealant microorganism, the entire Sporosarcina pasteurii urease gene sequences including ureA, ureB, ureC, ureD, ureE, ureF, and ureG from plasmid pBU11 were sub-cloned into the shuttle vector, pUCP18. The newly constructed plasmid, pUBU1, was transformed into two Pseudomonas aeruginosa strains, 8821 and PAO1, to develop recombinants capable of inducing calcite precipitation in addition to their own ability to produce EPS. Nickel-dependent urease activities were expressed from the recombinant P. aeruginosa 8821 (pUBU1) and P. aeruginosa PAO1 (pUBU1), at 99.4% and 60.9% of the S. pasteurii urease activity, respectively, in a medium containing 2mM NiCl2. No urease activities were detected from the wild type P. aeruginosa 8821 and P. aeruginosa PAO1 under the same growth conditions. Recombinant Pseudomonas strains induced CaCO3 precipitation at a comparable rate as S. pasteurii and scanning electron microscopy evidenced the complex of CaCO3 crystals and EPS layers surrounding the cells. The engineered strains produced in this study are expected to serve as a valuable reference to future biosealants that could be applied in the environment. However, the pathogenic potential of P. aeruginosa, used here only as a model system to show the proof of principle, prevents the use of this recombinant organism as a biosealant. In practical applications, other recombinant organisms should be used.

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.

The role of alginate in Pseudomonas aeruginosa EPS adherence, viscoelastic properties and cell attachment

Among various functions, extracellular polymeric substances (EPS) provide microbial biofilms with mechanical stability and affect initial cell attachment, the first stage in the biofilm formation process. The role of alginate, an abundant polysaccharide in Pseudomonas aeruginosa biofilms, in the viscoelastic properties and adhesion kinetics of EPS was analyzed using a quartz crystal microbalance with dissipation (QCM-D) monitoring technology. EPS was extracted from two P. aeruginosa biofilms, a wild type strain, PAO1, and a mucoid strain, PAOmucA22 that over-expresses alginate production. The higher alginate content in the EPS originating from the mucoid biofilms was clearly shown to increase both the rate and the extent of attachment of the EPS, as well as the layer's thickness. Also, the presence of calcium and elevated ionic strength increased the thickness of the EPS layer. Dynamic light scattering (DLS) showed that the presence of calcium and elevated ionic strength induced intermolecular attractive interactions in the mucoid EPS molecules. For the wild type EPS, in the presence of calcium, an elevated shift in the distribution of the diffusion coefficients was observed with DLS due to a more compacted conformation of the EPS molecules. Moreover, the alginate over-expression effect on EPS adherence was compared to the effect of alginate over-expression on P. aeruginosa cell attachment. In a parallel plate flow cell, under similar hydraulic and aquatic conditions as those applied for the EPS adsorption tests in the QCM-D flow cell, reduced adherence of the mucoid strain was clearly observed compared to the wild type isogenic bacteria. The results suggest that alginate contributes to steric hindrance and shielding of cell surface features and adhesins that are known to promote cell attachment.

Generalized Relationship between Numbers of Bacteria and Their Viability in Biofilms

Bacterial biofilms are confined communities that are encapsulated in protective layers of extracellular polymeric substances. Microscopic evaluation of biofilms of diverse bacterial strains on various substrata reveals that, in general, the percentage of viable bacteria decreases with the total number of bacteria in a biofilm.

Influence of EPS isolated from thermophilic sulphate-reducing bacteria on carbon steel corrosion

Extracellular polymeric substances (EPS) were isolated by centrifugation of thermophilic sulphate-reducing bacteria (SRB) grown in API-RP38 culture medium. The protein and polysaccharide fractions were quantified and the highest concentrations were extracted from a 14-day old culture. The effect of EPS on carbon steel corrosion was investigated by electrochemical techniques. At 30°C, a small amount of EPS in 3% NaCl solution inhibited corrosion, whilst excessive amounts of EPS facilitated corrosion. In addition, the inhibition efficiency of EPS decreased with temperature due to thermal desorption of the EPS. The results suggest that adsorbed EPS layers could be beneficial to anti-corrosion by hindering the reduction of oxygen. However, the accumulation of an EPS film could stimulate the anodic dissolution of the underlying steel by chelation of Fe2+ ions.

A mathematical model of quorum sensing regulated EPS production in biofilm communities

BackgroundBiofilms are microbial communities encased in a layer of extracellular polymeric substances (EPS). The EPS matrix provides several functional purposes for the biofilm, such as protecting bacteria from environmental stresses, and providing mechanical stability. Quorum sensing is a cell-cell communication mechanism used by several bacterial taxa to coordinate gene expression and behaviour in groups, based on population densities.ModelWe mathematically model quorum sensing and EPS production in a growing biofilm under various environmental conditions, to study how a developing biofilm impacts quorum sensing, and conversely, how a biofilm is affected by quorum sensing-regulated EPS production. We investigate circumstances when using quorum-sensing regulated EPS production is a beneficial strategy for biofilm cells.ResultsWe find that biofilms that use quorum sensing to induce increased EPS production do not obtain the high cell populations of low-EPS producers, but can rapidly increase their volume to parallel high-EPS producers. Quorum sensing-induced EPS production allows a biofilm to switch behaviours, from a colonization mode (with an optimized growth rate), to a protection mode.ConclusionsA biofilm will benefit from using quorum sensing-induced EPS production if bacteria cells have the objective of acquiring a thick, protective layer of EPS, or if they wish to clog their environment with biomass as a means of securing nutrient supply and outcompeting other colonies in the channel, of their own or a different species.

In situ imaging of Sulfobacillus thermosulfidooxidans on pyrite under conditions of variable pH using tapping mode atomic force microscopy

Sulfobacillus thermosulfidooxidans cells attached to pyrite surfaces were imaged using in situ tapping mode atomic force microscopy (AFM). The attached bacteria were dispersed in small groups without preferred orientation or site specific preference. The importance of extracellular polymeric substances (EPS) production by cells upon changing pH was examined. Bacteria detached or were dislodged and no EPS was detected for a rapid decrease from pH 2.2 to pH 1.0 in the solution. In contrast, a layer of EPS with increasing thickness was observed for a decrease from pH 2.2 to pH 1.4. Upon further acidification (pH 0.9) the cells were not dislodged but the thickness of the EPS layer decreased immediately, which is attributed to structural re-arrangements of the polymeric chains within the EPS network. This suggests that the bacteria respond to stress caused by increased acidity by expressing EPS, which acts as a buffer against unfavourable conditions in the environment. Secondary mineral formation (‘scale’) on the pyrite surface was found to form only in the presence of microorganisms and eventually covered the pyrite surface. Bacterial cells did not attach to the scale surface. Upon scale removal, etch pits with similar shape to bacterial cells were observed in the underlying pyrite surface.

Extracellular Polymeric Substances (EPS) in a Hybrid Growth Membrane Bioreactor (HG-MBR): Viscoelastic and Adherence Characteristics

Extracellular polymeric substances (EPS) comprising the microbial biofilms in membrane bioreactor (MBR) systems are considered the most significant factor affecting sludge viscoelastic properties as well as membrane fouling. Understanding the water chemistry effects on EPS viscoelastic, conformational, and adherence properties are critical for defining the microbial biofilm's propensity of fouling the membrane surface. In this study, EPS extracted from a hybrid growth membrane bioreactor (HG-MBR) were analyzed for their adherence, viscoelastic properties and size distribution using quartz crystal microbalance with dissipation monitoring (QCM-D) and dynamic light scattering (DLS), respectively. Also, adsorption characteristics of EPS extracted from different locations in the HG-MBR (bioreactor liquor, fluidized carriers, and membrane surface) were defined and linked to the extent of the total polysaccharide content in the EPS. In accordance with the Derjaguin-Landau-Verwey-Overbeek (DLVO) theory, more EPS were adsorbed at higher ionic strength, lower pH and in the presence of calcium cations. Based on the QCM-D results, the calculated thickness of the EPS adsorbed layer was increased at lower ionic strength, higher pH, and only had a minor increase in the presence of calcium cations. The calculated shear modules and shear viscosity suggest that at lower pH and in the presence of calcium, EPS becomes more viscous and elastic, respectively. DLS analysis correlated to the QCM-D results: A decrease in the hydrodynamic radius of the EPS colloids was observed at lower pH, and in the presence of calcium, most likely attributed to intermolecular attraction forces. Based on this study, low pH and presence of calcium may induce flocs' stability that resist erosion in the MBRs, while on the other hand, these conditions may induce the formation of an elastic and viscous EPS layer fouling the ultrafiltration (UF) membrane.

The effect of ionic strength on the electrophoretic mobility and protonation constants of an EPS-producing bacterial strain

The production of extracellular polymeric substances (EPS) by bacteria significantly alters the physical structure of the cell wall-solution interface, potentially affecting cell surface reactivity and colloidal (transport) properties of the cells. This study investigated the effect of the EPS layer on the electrostatic properties of the cell surface by conducting electrophoretic mobility (EM) analysis and potentiometric titrations at ionic strengths of 0.001 M, 0.01 M and 0.1 M, using both native cells, with the EPS-layer intact, and EPS-free cells of the gram-positive thermophile, Bacillus licheniformis S-86. Electrophoretic mobility measurements indicated that the isoelectric point for both cell types was below pH 2. At low ionic strength (0.001 M), expansion of the EPS layer causes the EM of the native cells to have a smaller magnitude than that of the EPS-free cells. As ionic strength increases, this effect diminishes and at 0.1 M the native cells have a higher EM than the EPS-free cells, which indicates that there is a relatively high charge density in the EPS layer. Higher total site concentrations obtained using potentiometric titration data modelling for the native cells are in agreement with the electrophoretic mobility data, and indicate a higher proton-active site density in the EPS layer compared to the EPS-free cell wall. The results of data modelling for titrations conducted at all three ionic strength values showed that there are no systematic changes in deprotonation constants or site concentrations as a function of ionic strength, indicating that the use of a non-electrostatic model is justified for this EPS-producing bacterial strain.

Mechanism of bioleaching chalcopyrite by Acidithiobacillus ferrooxidans in agar-simulated extracellular polymeric substances media

The mechanism of leaching chalcopyrite by Acidithiobacillus ferrooxidans (A. ferrooxidans) in agar-simulated extracellular polymeric substances (EPS) media was investigated. The results indicate that bacterial EPS can release H+ and concentrate Fe3+; Fe2+ is movable between agar-simulated EPS phase and bulk solution phase, but it is difficult for Fe3+ to move due to its hydroxylation and EPS complex action; A. ferrooxidans first prefer Fe2+ as energy to metabolize compared with chalcopyrite, and a suitable simulated EPS environment for bacterial living is at about pH 1.8; the iron precipitates and jarosites formed by a lot of biologically oxidized Fe3+ cover the simulated EPS easily and form an impermeable deposit acting as a limited barrier of ion transport that attenuates the aggressiveness of the bioleaching attack. The EPS layer blocked by iron precipitates or jarosites is responsible for the chalcopyrite passivation.

Role of Extracellular Polymeric Substances (EPS) in Biofouling of Reverse Osmosis Membranes

This study elucidates the mechanisms by which extracellular polymeric substances (EPS) impact permeate water flux and salt rejection during biofouling of reverse osmosis (RO) membranes. RO fouling experiments were conducted with Pseudomonas aeruginosa PAO1, EPS extracted from PAO1 biofilms, and dead PAO1 cells fixed in formaldehyde. While a biofouling layer of dead bacterial cells decreases salt rejection and permeate flux by a biofilm-enhanced osmotic pressure mechanism, the EPS biofouling layer adversely impacts permeate flux by increasing the hydraulic resistance to permeate flow. During controlled fouling experiments with extracted EPS in a simulated wastewater solution, polysaccharides adsorbed on the RO membranes much more effectively than proteins (adsorption efficiencies of 61.2-88.7% and 11.6-12.4% for polysaccharides and proteins, respectively). Controlled fouling experiments with EPS in sodium chloride solutions supplemented with 0.5 mM calcium ions (total ionic strength of 15 mM) indicate that calcium increases the adsorption efficiency of polysaccharides and DNA by 2- and 3-fold, respectively. The increased adsorption of EPS onto the membrane resulted in a significant decrease in permeate water flux. Corroborating with these calcium effects, atomic force microscopy (AFM) measurements demonstrated that addition of calcium ions to the feed solution results in a marked increase in the adhesion forces between a carboxylated particle probe and the EPS layer. The increase in the interfacial adhesion forces is attributed to specific EPS-calcium interactions that play a major role in biofouling of RO membranes.

Effect of water-swollen organic materials on crossflow filtration performance

Studies have demonstrated that cakes formed on filter media when filtering microorganisms behave differently from cakes of inorganic particles. It has been suggested that this is due to the water-swollen layer of extracellular polymeric substances on the cell walls of the microorganisms. This idea was tested by preparing simple core–shell particles with a hard polystyrene core and a water-swollen polyacrylic acid shell. The same filtration phenomena were observed when crossflow filtering core–shell particles as reported for microorganisms, i.e., a compressible cake was formed, the compression was reversible, and a linear dependency between resistance and pressure was observed. Thus, the water-swollen layer greatly affected the filtration. From the slope obtained by plotting resistance vs. pressure, the pressure required to double the initial resistance was calculated. This pressure was used as a measure of cake compressibility and determined to be 0.35 bar, in the same range as reported for the microorganisms.

Bacterial extracellular polymeric substances (EPS) mediate CaCO 3 morphology and polymorphism

The effect of bacterial extracellular polymers (EPS) on calcium carbonate morphology and polymorphism was investigated by means of free-drift mineralisation experiments, conducted using cells of the EPS-producing thermotolerant Bacillus licheniformis S-86. The cells either had the EPS layer intact (native cells) or had the EPS layer removed (EPS-free cells). Experiments were also undertaken in the absence of cells, but with the addition of the extracted EPS solution. It was found that the abiotic control and EPS-free cell experiments contained vaterite and calcite when sampled after 12 h and 48 h, whereas the native cell and EPS-solution experiments contained only calcite. These results suggest that the presence of EPS inhibits vaterite precipitation. It is proposed that dissolved organic carbon (DOC) released from the EPS complexes Ca 2+ ions in solution, reducing the calcium carbonate saturation and favouring calcite precipitation over vaterite. After 7 days, all experiments contained only calcite. However, distinct morphological differences between the calcite crystals precipitated in the four experiments were preserved after a period of 7 days, suggesting that crystal morphology may indicate the biological conditions under which calcite precipitated.

Atomic Force Microscopy of Mechanically Trapped Bacterial Cells

This article presents a study on the influence of the protocol used for immobilization of bacterial cells onto surfaces by mechanically trapping them into a filter. In this sense, the surface and structure of trapped cells are analyzed. Bacteria can be present solely or with extracellular polymeric substances (EPS). To test the behavior of the EPS layer duing the filtering process, different strains of a well-known EPS-producer bacteria (Staphylococcus epidermidis), which produce an extracellular matrix clearly visible in AFM images, have been used. Results show that this immobilization method can cause severe structural and mechanical deformation to the cell membrane. This altered mechanical state may possibly influence the parameters derived from AFM force curves (which are micro/nano-mechanical tests). Also, our results suggest that the EPS layer might move during the filtering process and could accumulate at the upper part of the cell, thus favoring distorted data of adhesion/pull-off forces as measured by an AFM tip, especially in the case of submicron-sized microbial cells such as bacteria.

Interaction between Calcite and Phosphorus in Biomineralization Processes in Tufa Carbonates

Using electron microscopy techniques (SEM, LTSEM) coupled with analytical methods (XRD and EDS) the role of phosphorus has been assessed in the formation of freshwater calcite deposits (tufa) in a small pond of the Ruidera Lakes (Spain). Differences between the cell walls and sheaths of bacteria and eukaryotic algae as well as the existence of additional layers of extracellular polymeric substances (EPS) were features that lead to differences in the process of induced calcite biomineralization. Phosphorus has influence in the biomineralization of the EPS, sheaths and cell walls of cyanobacteria allowing for fossil preservation whereas does not participate in the calcite precipitation around algae and mosses. This variability may explain the different positive or negative roles played by natural or artificial inputs of phosphorus in hard water lakes and the different morphological features of calcite precipitates associated with eukaryotic and cyanobacteria picoplankton found in natural environments. The biomineralization observed is in agreement with the isotopic composition of the tufa layers that reflect the variations in environmental conditions around biological communities. (© 2006 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim)