Microphytobenthos,their Extracellular Polymeric Substances,and the Morphogenesis of Intertidal Sediments

Intertidal sediments are important areas that separate the land from the sea and form natural coastal defenses. They are known as highly productive ecosystems, fueling the coastal food web. It is also conceived that microphytobenthos contribute to the stability of intertidal sediments by increasing the erosion threshold and that they are major players in coastal morphodynamics. Depending on the sedimentary composition of intertidal flats, different types of microphytobenthos colonize the sediment surface. Fine sand sediment is often colonized by cyanobacteria, prokaryotic algae, which form dense and rigid microbial mats. Mudflats on the other hand are characterized by the development of thin biofilms of epipelic diatoms. Both groups of phototrophic microorganisms excrete extracellular polymeric substances (EPS), but they do so in different ways and for different reasons. Two operationally defined fractions, water- and EDTA-extractable EPS, have been obtained from intertidal diatom biofilms and from cultures. They differ in composition and their production seems to be under different metabolic control. Water-extractable EPS are considered to be closely associated with the diatoms and are rich in neutral sugars, notably glucose. These EPS show a dynamic relationship with the microphytobenthic biomass. EDTA-extractable EPS are tightly bound to the sediment, probably through bridging by divalent ions. This material is rich in uronic acids and other acid sugars and is weakly related to chlorophyll. These EPS have been conceived to be a major factor in the structuring and diagenesis of coastal sediments and essential for increasing the sediment erosion threshold. However, this relationship is now questioned.     

Bacteria Associated with Benthic Diatoms from Lake Constance: Phylogeny and Influences on Diatom Growth and Secretion of Extracellular Polymeric Substances

The composition of diatom-associated bacterial communities was studied with 14 different unialgal xenic diatom cultures isolated from freshwater epilithic biofilms of Lake Constance, Germany. A clear dominance of Alphaproteobacteria was observed, followed by Betaproteobacteria, Gammaproteobacteria, Bacteroidetes, and Verrucomicrobia. Pure cultures of the diatom Cymbella microcephala, which was found to be dominant in epilithic biofilms in Lake Constance, were cocultivated with six associated bacterial strains. All these bacterial strains were able to grow in C. microcephala cultures in the absence of organic cosubstrates. Diatom growth was generally enhanced in the presence of bacteria, and polysaccharide secretion was generally increased in the presence of Proteobacteria. The monomer composition of extracellular polysaccharides of C. microcephala changed in relation to the presence of different bacteria, but the dominant monomers were less affected. Our results indicate that these changes were caused by the diatom itself rather than by specific bacterial degradation. One Bacteroidetes strain strongly influenced carbohydrate secretion by the alga via extracellular soluble compounds. Biofilms were formed only in the presence of bacteria. Phylogenetic analysis and coculture studies indicate an adaptation of Proteobacteria and Bacteroidetes to the microenvironment created by the diatom biofilm.     

Interactions between Benthic Copepods,Bacteria and Diatoms Promote Nitrogen Retention in Intertidal Marine Sediments

The present study aims at evaluating the impact of diatoms and copepods on microbial processes mediating nitrate removal in fine-grained intertidal sediments. More specifically, we studied the interactions between copepods, diatoms and bacteria in relation to their effects on nitrate reduction and denitrification. Microcosms containing defaunated marine sediments were subjected to different treatments: an excess of nitrate, copepods, diatoms (Navicula sp.), a combination of copepods and diatoms, and spent medium from copepods. The microcosms were incubated for seven and a half days, after which nutrient concentrations and denitrification potential were measured. Ammonium concentrations were highest in the treatments with copepods or their spent medium, whilst denitrification potential was lowest in these treatments, suggesting that copepods enhance dissimilatory nitrate reduction to ammonium over denitrification. We hypothesize that this is an indirect effect, by providing extra carbon for the bacterial community through the copepods'' excretion products, thus changing the C/N ratio in favour of dissimilatory nitrate reduction. Diatoms alone had no effect on the nitrogen fluxes, but they did enhance the effect of copepods, possibly by influencing the quantity and quality of the copepods'' excretion products. Our results show that small-scale biological interactions between bacteria, copepods and diatoms can have an important impact on denitrification and hence sediment nitrogen fluxes.     

Microbial Extracellular Polymeric Substances (EPS) in Fresh Water Sediments

Microbially produced extracellular polymeric substances (EPS) have been linked with many important ecological functions in natural sediments; yet, most information has been derived from marine systems. The present paper is the first comprehensive study on EPS (i.e., carbohydrates and proteins) dynamics in riverine sediments addressing spatial (six reservoirs and four groyne fields across three European rivers), temporal (all seasons in 2003–2005), and vertical (over a 50-cm sediment depth transect) pattern. The variation in hydrodynamic regime found in the reservoirs and groyne fields was reflected in the biomass and composition of the benthic microorganisms that produce EPS. The microphytobenthic communities consisted mainly of diatoms and a higher algal biomass (up to 248 μg g⁻¹ dry weight, DW) seemed to be indicative for higher amounts of secreted colloidal carbohydrates. Consequently, the model proposed by Underwood and Smith (1998) for the relation chlorophyll–colloidal carbohydrates was also applicable for upper riverine sediment layers. The close relation between algal biomass and bacterial cell counts (10⁸–10⁹ cells g⁻¹ DW) supports the idea of bacterial use of the secreted EPS. However, the data also suggest a contribution to the EPS pool through bacterial secretion of proteins/extracellular enzymes and possibly carbohydrates. Over depth, the relationships between microorganisms and EPS became increasingly decoupled along with increasing ratios of bound (refractory) to colloidal (labile) EPS. These data suggest fresh production of polymeric substances in upper sediment layers and mainly accumulation of refractory, biodegraded material in deeper layers. The high contents of EPS colloidal and bound carbohydrates (0.1–1.8 and 1.3–6.7 mg g⁻¹ DW, respectively) and EPS proteins (0.4–12.9 mg g⁻¹ DW) at the freshwater study sites might indicate an important role in sediment ecology.     

Nanoscale Observations of Extracellular Polymeric Substances Deposition on Phyllosilicates by an Ectomycorrhizal Fungus

Microorganisms colonizing surfaces can exude a wide range of substances, generally called Extracellular Polymeric Substances (EPS). While EPS has often been visualized as thick mature strata embedding microbes, the initial phases of EPS production, its structure at the micro- and nanoscale and the microbial wall areas involved in its exudation are less known. In this work we use Atomic Force Microscopy to image EPS produced by the fungus Paxillus involutus on phyllosilicate surfaces. Hyphal tips initially deposit EPS which assumes the shape of a “halo” surrounding hyphae. The fusion of adjacent EPS halos is likely responsible for the creation of EPS monolayers covering mineral surfaces. It is also proposed that a specific region of hyphae initiates the formation of mineral channels produced by fungi. The results presented here permit for the first time to propose a model for the initial stages of EPS accumulation in fungi and filamentous microorganisms in general.     

Extraction of Structural Extracellular Polymeric Substances from Aerobic Granular Sludge

To evaluate and develop methodologies for the extraction of gel-forming extracellular polymeric substances (EPS), EPS from aerobic granular sludge (AGS) was extracted using six different methods (centrifugation, sonication, ethylenediaminetetraacetic acid (EDTA), formamide with sodium hydroxide (NaOH), formaldehyde with NaOH and sodium carbonate (Na₂CO₃) with heat and constant mixing). AGS was collected from a pilot wastewater treatment reactor. The ionic gel-forming property of the extracted EPS of the six different extraction methods was tested with calcium ions (Ca²⁺). From the six extraction methods used, only the Na₂CO₃ extraction could solubilize the hydrogel matrix of AGS. The alginate-like extracellular polymers (ALE) recovered with this method formed ionic gel beads with Ca²⁺. The Ca²⁺-ALE beads were stable in EDTA, formamide with NaOH and formaldehyde with NaOH, indicating that ALE are one part of the structural polymers in EPS. It is recommended to use an extraction method that combines physical and chemical treatment to solubilize AGS and extract structural EPS.     

Dynamic Membrane Formation in Anaerobic Dynamic Membrane Bioreactors: Role of Extracellular Polymeric Substances

Dynamic membrane (DM) formation in dynamic membrane bioreactors plays an important role in achieving efficient solid-liquid separation. In order to study the contribution of extracellular polymeric substances (EPS) to DM formation in anaerobic dynamic membrane bioreactor (AnDMBR) processes, EPS extraction from and re-addition to bulk sludge were carried out in short-term filtration tests. DM formation behaviors could be well simulated by cake filtration model, and sludge with EPS re-addition showed the highest resistance coefficient, followed by sludge after EPS extraction. The DM layers exhibited a higher resistance and a lower porosity for the sludge sample after EPS extraction and for the sludge with EPS re-addition. Particle size of sludge flocs decreased after EPS extraction, and changed little with EPS re-addition, which was confirmed by interaction energy analysis. Further investigations by confocal laser scanning microscopy (CLSM) analysis and batch tests suggested that the removal of in-situ EPS stimulated release of soluble EPS, and re-added EPS were present as soluble EPS rather than bound EPS, which thus improved the formation of DM. The present work revealed the role of EPS in anaerobic DM formation, and could facilitate the operation of AnDMBR processes.     

The Fate of Nitrate in Intertidal Permeable Sediments

Coastal zones act as a sink for riverine and atmospheric nitrogen inputs and thereby buffer the open ocean from the effects of anthropogenic activity. Recently, microbial activity in sandy permeable sediments has been identified as a dominant source of N-loss in coastal zones, namely through denitrification. Some of the highest coastal denitrification rates measured so far occur within the intertidal permeable sediments of the eutrophied Wadden Sea. Still, denitrification alone can often account for only half of the substantial nitrate (NO₃ ⁻) consumption. Therefore, to investigate alternative NO₃ ⁻ sinks such as dissimilatory nitrate reduction to ammonium (DNRA), intracellular nitrate storage by eukaryotes and isotope equilibration effects we carried out ¹⁵NO₃ ⁻ amendment experiments. By considering all of these sinks in combination, we could quantify the fate of the ¹⁵NO₃ ⁻ added to the sediment. Denitrification was the dominant nitrate sink (50–75%), while DNRA, which recycles N to the environment accounted for 10–20% of NO₃ ⁻ consumption. Intriguingly, we also observed that between 20 and 40% of ¹⁵NO₃ ⁻ added to the incubations entered an intracellular pool of NO₃ ⁻ and was subsequently respired when nitrate became limiting. Eukaryotes were responsible for a large proportion of intracellular nitrate storage, and it could be shown through inhibition experiments that at least a third of the stored nitrate was subsequently also respired by eukaryotes. The environmental significance of the intracellular nitrate pool was confirmed by in situ measurements which revealed that intracellular storage can accumulate nitrate at concentrations six fold higher than the surrounding porewater. This intracellular pool is so far not considered when modeling N-loss from intertidal permeable sediments; however it can act as a reservoir for nitrate during low tide. Consequently, nitrate respiration supported by intracellular nitrate storage can add an additional 20% to previous nitrate reduction estimates in intertidal sediments, further increasing their contribution to N-loss.     

Influences of Extracellular Polymeric Substances on the Dewaterability of Sewage Sludge during Bioleaching

Extracellular polymeric substances (EPS) play important roles in regulating the dewaterability of sludge. This study sought to elucidate the influence of EPS on the dewaterability of sludge during bioleaching process. Results showed that, in bioleaching system with the co-inoculation of Acidithiobacillus thiooxidans TS6 and Acidithiobacillus ferrooxidans LX5 (A. t+A. f system), the capillary suction time (CST) of sludge reduced from 255.9 s to 25.45 s within 48 h, which was obviously better than the controls. The correlation analysis between sludge CST and sludge EPS revealed that the sludge EPS significantly impacted the dewaterability of sludge. Sludge CST had correlation with protein content in slime and both protein and polysaccharide contents in TB-EPS and Slime+LB+TB layers, and the decrease of protein content in slime and decreases of both protein and polysaccharide contents in TB-EPS and Slime+LB+TB layers improved sludge dewaterability during sludge bioleaching process. Moreover, the low sludge pH (2.92) and the increasing distribution of Fe in the solid phase were another two factors responsible for the improvement of sludge dewaterability during bioleaching. This study suggested that during sludge bioleaching the growth of Acidithiobacillus species resulted in the decrease of sludge pH, the increasing distribution of Fe in the solid phase, and the decrease of EPS content (mainly including protein and/or polysaccharide) in the slime, TB-EPS, and Slime+LB+TB layers, all of which are helpful for sludge dewaterability enhancement.     

Linking Intertidal and Subtidal Food Webs: Consumer-Mediated Transport of Intertidal Benthic Microalgal Carbon

We examined stable carbon and nitrogen isotope ratios for a large variety of consumers in intertidal and subtidal habitats, and their potential primary food sources [i.e., microphytobenthos (MPB), phytoplankton, and Phragmites australis] in a coastal bay system, Yeoja Bay of Korea, to test the hypothesis that the transfer of intertidal MPB-derived organic carbon to the subtidal food web can be mediated by motile consumers. Compared to a narrow δ¹³C range (−18 to −16‰) of offshore consumers, a broad δ¹³C range (−18 to −12‰) of both intertidal and subtidal consumers indicated that ¹³C-enriched sources of organic matter are an important trophic source to coastal consumers. In the intertidal areas, δ¹³C of most consumers overlapped with or was ¹³C-enriched relative to MPB. Despite the scarcity of MPB in the subtidal, highly motile consumers in subtidal habitat had nearly identical δ¹³C range with many intertidal foragers (including crustaceans and fish), overlapping with the range of MPB. In contrast, δ¹³C values of many sedentary benthic invertebrates in the subtidal areas were similar to those of offshore consumers and more ¹³C-depleted than motile foragers, indicating high dependence on phytoplankton-derived carbon. The isotopic mixing model calculation confirms that the majority of motile consumers and also some of subtidal sedentary ones depend on intertidal MPB for more than a half of their tissue carbon. Finally, although further quantitative estimates are needed, these results suggest that direct foraging by motile consumers on intertidal areas, and thereby biological transport of MPB-derived organic carbon to the subtidal areas, may provide important trophic connection between intertidal production and the nearshore shallow subtidal food webs.     

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-μm 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.     

Evaluation of Fluorescently Labeled Lectins for Noninvasive Localization of Extracellular Polymeric Substances in Sphingomonas Biofilms

Three strains of Sphingomonas were grown as biofilms and tested for binding of five fluorescently labeled lectins (Con A-type IV-TRITC or -Cy5, Pha-E-TRITC, PNA-TRITC, UEA 1-TRITC, and WGA-Texas red). Only ConA and WGA were significantly bound by the biofilms. Binding of the five lectins to artificial biofilms made of the commercially available Sphingomonas extracellular polysaccharides was similar to binding to living biofilms. Staining of the living and artificial biofilms by ConA might be explained as binding of the lectin to the terminal mannosyl and terminal glucosyl residues in the polysaccharides secreted by Sphingomonas as well as to the terminal mannosyl residue in glycosphingolipids. Staining of the biofilms by WGA could only be explained as binding to the Sphingomonas glycosphingolipid membrane, binding to the cell wall, or nonspecific binding. Glycoconjugation of ConA and WGA with the target sugars glucose and N-acetylglucosamine, respectively, was used as a method for evaluation of the specificity of the lectins towards Sphingomonas biofilms and Sphingomonas polysaccharides. Our results show that the binding of lectins to biofilms does not necessarily prove the presence of specific target sugars in the extracellular polymeric substances (EPS) in biofilms. The lectins may bind to non-EPS targets or adhere nonspecifically to components of the biofilm matrix.     

Migratory Responses of Benthic Diatoms to Light and Temperature Monitored by Chlorophyll Fluorescence

As an important adaptation for survival in the sediments of intertidal flats, benthic diatoms move up and down in response to a wide range of environmental stimuli. We investigated the vertical migration of two diatoms—Cylindrotheca closterium (Agradh) Kützing (B-25) and Nitzschia sp. (B-3)—under different combinations of light intensity and temperature conditions. An imaging pulse amplitude modulated (PAM) fluorometer was used to measure the minimum fluorescence (F ₀) in order to monitor variations in diatom biomass in surface sediments. Rapid light curves (RLCs) were applied to assess their photosynthetic activities. Both species had increased motility under higher temperatures, with the longer valved C. closterium being twice as fast as the shorter valved Nitzschia sp. The former was also influenced by exposure to light intensities of 100 or 250 μmol m⁻² s⁻¹, whereas the latter was not. Consequently, no light/temperature interaction effect was associated with the vertical migration of Nitzschia sp., perhaps because of its lower photosynthetic capacity and smaller cell size. Therefore, we conclude that motile benthic diatoms exhibit species-specific responses to light and temperature due to differences in their photosynthetic capability and morphological characteristics.     

The Impact of Artisanal Fishery on a Tropical Intertidal Benthic Fish Community

We examined the benthic fishes and artisanal fishery in the intertidal flats of Inhaca Island, Mozambique. Results of a questionnaire indicated that catches had decreased, and that piscivorous fish have disappeared. Results of a catch sampling study indicated that current catch rates are low, < 2 kg person^–1 fishing trip^–1. Use of fishing gear was significantly related to season, diel and lunar tidal phase, and habitat. Forty-eight fish species were observed in the catches with eight species comprising 80% of the catch of 1814 specimens. The annual catch was estimated at 26.2t for the whole bay. Highest fishing pressure was observed in the central section of the bay. A demersal fish survey was carried out with a 2-m beam trawl to sample the fish community. Two different areas were sampled, one area with a low, and one with a high fishing pressure. A total of 19889 fishes were caught comprising 93 species. Gobies dominated the catches and accounted for 56% of all specimens. Fishes were small with a mean standard length of 29mm. The Saco area exhibited the highest catch rates and biomass (maximum of 1040 fish 1000 m^–2 and 1490g 1000 m^–2), and the highest species richness and evenness values. Catch composition was different between the two sampling areas, and was strongly affected by season, but less by habitat. Total fish biomass was estimated at 5.6t for the whole area. Stomach content varied with habitat, and season, and was dominated by benthic invertebrates. The largest estimates of consumption were obtained in the tidal channel and the Zostera beds. Mean consumption of benthic organisms was 1.3g AFDW m^–2 yr^–1. The area seemed to be overfished. The heavily fished areas exhibited lower catch rates, lower proportion of piscivorous fish, increased proportion of small fish, and a decrease in species diversity.     

Isolation of New Chlorosis-inducing Substances Produced by Fungus

Five analogs of the insecticidal lignan of the Phryma were synthesized by modifying its 3,4-methylenedioxyphenyl group with a phenyl, 4-methoxyphenyl, 3-methoxyphenyl, 3,4-dimethoxyphenyl, or 4-chlorophenyl group to evaluate the contribution of the methylenedioxy function to the activity. The assay results revealed that the 4-oxymethylene part of the 3,4-methylenedioxyphenyl group collaborated more for strengthening the insecticidal activity than the 3-oxymethylene part did. Unexpectedly, the 3,4-dimethoxyphenyl analog was totally inactive, even at a high dose level.     

Quantitative Importance,Composition, and Seasonal Dynamics of Protozoan Communities in Polyhaline versus Freshwater Intertidal Sediments

The quantitative importance and composition of protozoan communities was investigated in sandy and silty intertidal sediments of a polyhaline and a freshwater site in the Schelde estuary. Total biomass of the protozoans studied, integrated over the upper 4 cm of the sediment, ranged from 41 to 597 mg C m^–2 and was in the same order of magnitude at the polyhaline and the freshwater intertidal site. Nanoheterotrophs were the dominant protozoans, in terms of both abundance and biomass. Ciliate abundances appeared to be largely determined by physical constraints, namely, the amount of interstitial space and hydrodynamic disturbances. It remains unclear which factors control nanoheterotrophic abundances and biomasses, which showed comparatively little seasonal and between-site fluctuations. Salinity differences were clearly reflected in the protozoan community composition. The dominant role of sessile ciliates is a unique feature of sediments in the freshwater tidal reaches, which can be attributed to the dynamic nature of sedimentation and resuspension processes associated with the maximum turbidity zone. Based on biomass ratios and estimated weight-specific metabolic rates, protozoa possibly accounted for ~29 to 96% of the estimated combined metabolic rate of protozoan and metazoan consumers at our sampling stations in late spring/early autumn. The contribution of protozoa to this combined metabolic rate was higher at the sandy than at the silty stations and was mainly accounted for by the nanoheterotrophs. These data emphasize the potential importance of small protozoa in sediments and suggest that protozoa are important components of benthic food webs.     

Role of Extracellular Polymeric Substances in the Surface Chemical Reactivity of Hymenobacter aerophilus,a Psychrotolerant Bacterium

Bacterial surface layers, such as extracellular polymeric substances (EPS), are known to play an important role in metal sorption and biomineralization; however, there have been very few studies investigating how environmentally induced changes in EPS production affect the cell''s surface chemistry and reactivity. Acid-base titrations, cadmium adsorption assays, and Fourier transform infrared spectroscopy (FT-IR) were used to characterize the surface reactivities of Hymenobacter aerophilus cells with intact EPS (WC) or stripped of EPS (SC) and purified EPS alone. Linear programming modeling of titration data showed SC to possess functional groups corresponding to phosphoryl (pK_a ∼6.5), phosphoryl/amine (pK_a ∼7.9), and amine/hydroxyl (pK_a ∼9.9). EPS and WC both possess carboxyl groups (pK_a ∼5.1 to 5.8) in addition to phosphoryl and amine groups. FT-IR confirmed the presence of polysaccharides and protein in purified EPS that can account for the additional carboxyl groups. An increased ligand density was observed for WC relative to that for SC, leading to an increase in the amount of Cd adsorbed (0.53 to 1.73 mmol/liter per g [dry weight] and 0.53 to 0.59 mmol/liter per g [dry weight], respectively). Overall, the presence of EPS corresponds to an increase in the number and type of functional groups on the surface of H. aerophilus that is reflected by increased metal adsorption relative to that for EPS-free cells.Acid-base titrations are frequently used to characterize microbial cell surface reactivity, in particular, the ability of the cell to adsorb and desorb protons (19, 21, 47). This ability is conferred by the presence of proton-reactive surface functional groups that are also responsible for the surface adsorption of other cations, including dissolved metals. Thus, a microbe''s ability to immobilize metals and influence metal transport is largely dependent on the nature of the reactive sites found at the cell-water interface, namely, their concentrations and chemical affinities (in terms of equilibrium surface stability constants) for cations such as protons and metals.Both Gram-negative and Gram-positive bacteria have been characterized extensively using acid-base titration to determine their reactivity with respect to geochemical processes (18, 22, 33). To date, most work has focused on mesophilic and strictly heterotrophic model organisms; however, some work has also been done with cyanobacteria (29, 44) and thermophiles (19, 47). While proton sorption assays provide information on surface site densities and acidity constants, a more direct assessment of a microbe''s ability to interact with aqueous metals is the metal adsorption assay, where a cell''s ability to adsorb metal ions from solution is measured over a range of pH values. Metal adsorption assays have been used to characterize microbes from a wide variety of environments to determine their potential for bioremediation of heavy metal contamination (21, 26), their influence on geochemical cycling (5, 16), and their ability to serve as nucleation sites for mineral authigenesis (3, 43). Although more than 80% of the Earth''s biosphere is cold (37), to our knowledge there have been no published studies of acid-base surface chemistry for psychrotolerant bacteria, although recent studies examining metal adsorption have been published (53, 54).Extracellular polymeric substances (EPS) are produced by both prokaryotes and eukaryotes in a wide variety of environments (15). Although the relative quantities of EPS components are highly variable, polysaccharides are usually dominant, with proteins and, to a lesser extent, nucleic acids and lipids also present (15). The production of EPS can be important in mediation of environmental interactions, such as adhesion to surfaces and aggregation (35, 49); mineral weathering (28, 51); microbial tolerance of toxic metals through sequestration of metal ions outside the cell (1, 11); and biomineralization (27). Indeed, the stability of metal-surface complexes is great enough to affect metal mobility in many aqueous systems (14), which can, in turn, affect the distribution of metals in the environment (32).The physical and chemical characteristics of EPS have usually been studied using cells with intact EPS or on purified EPS (7, 8, 17, 34, 41, 51). Interestingly, few studies have compared cells with and without surface layers such as EPS (44, 47), despite the fact that EPS and other external layers alter the cell surface presented to the environment, potentially changing both the type and the quantity of functional groups available for environmental interactions. Accordingly, the purpose of this study was to determine the changes in cell surface reactivity resulting from the production of EPS by Hymenobacter aerophilus, a psychrotolerant bacterium. Acid-base titrations and cadmium adsorption assays were used to compare the numbers and types of functional groups on the surfaces of bacterial cells presenting intact EPS and those from which EPS had been removed mechanically.     

Extracellular Polymeric Substances (EPS) of Freshwater Biofilms Stabilize and Modify CeO2 and Ag Nanoparticles

Streams are potential receiving compartments for engineered nanoparticles (NP). In streams, NP may remain dispersed or settle to the benthic compartment. Both dispersed and settling NP can accumulate in benthic biofilms called periphyton that are essential to stream ecosystems. Periphytic organisms excrete extracellular polymeric substances (EPS) that interact with any material reaching the biofilms. To understand the interaction of NP with periphyton it is therefore crucial to study the interaction of NP with EPS. We investigated the influence of EPS on the physicochemical properties of selected NP (CeO₂, Ag) under controlled conditions at pH 6, 7.6, 8.6 and light or dark exposure. We extracted EPS from five different periphyton communities, characterized the extracts, and exposed CeO₂ and carbonate-stabilized Ag NP (0.5 and 5 mg/L, both 25 nm primary particle size) and AgNO₃ to EPS (10 mg/L) over two weeks. We measured NP size distribution, shape, primary particle size, surface plasmon resonance, and dissolution. All EPS extracts were composed of biopolymers, building blocks of humic substances, low molecular weight (M_r) acids, and small amphiphilic or neutral compounds in varying concentrations. CeO₂ NP were stabilized by EPS independent of pH and light/dark while dissolution increased over time in the dark at pH 6. EPS induced a size increase in Ag NP in the light with decreasing pH and the formation of metallic Ag NP from AgNO₃ at the same conditions via EPS-enhanced photoreduction. NP transformation and formation were slower in the extract with the lowest biopolymer and low M_r acid concentrations. Periphytic EPS in combination with naturally varying pH and light/dark conditions influence the properties of the Ag and CeO₂ NP tested and thus the exposure conditions within biofilms. Our results indicate that periphytic organisms may be exposed to a constantly changing mixture of engineered and naturally formed Ag NP and Ag⁺.     

Composition of an Extracellular Polysaccharide Produced by Sphaerotilus natans

The capsular polysaccharide of Sphaerotilus natans has been isolated, purified, and analyzed. Chromatographic and chemical analyses performed on acid hydrolyzates of the purified material have shown that the major components are fucose, galactose, glucose, and glucuronic acid in approximately equimolar amounts. Glucose and glucuronic acid are believed to occur as an aldobiuronic acid unit.     

Plant Growth Substances Produced by Micro-organisms of Soil and Rhizosphere

S ummary : Micro-organisms isolated from rhizospheres and rhizoplanes of wheat plants, and from root-free soil, produced growth regulating substances with the properties of gibberellins and indolyl-3-acetic acid (IAA). Substances inhibiting extensions of pea plant internodes and lettuce hypocotyls were also produced, especially by bacteria from the root region of seedlings 6 days old. Bacteria producing growth promoting substances were most abundant on roots of older plants. Seedlings grown aseptically with added gibberellic acid (GA₃) and IAA, or grown with a soil inoculum, developed similarly and differed in their morphology from those grown aseptically without additives.