Excretion Of Extracellular Polymeric Substances Research Articles

Multispecies biofilms on reverse osmosis membrane dictate the function and characteristics of the bacterial communities rather than their structure

The main reason for the deterioration of membrane operation during water purification processes is biofouling, which has therefore been extensively studied. Biofouling was shown to reduce membrane performance reflected by permeate flux decline, reduced selectivity, membrane biodegradation, and consequently, an increase in energy consumption. Studies of biofouling focused on the identification of the assembled microbial communities, the excretion of extracellular polymeric substances (EPS), and their combined role in reduced membrane performance and lifetime. However, the link between the structure and function of biofouling communities has not been elucidated to date. Here, we provide a novel insight, suggesting that bacterial functions rather than composition control biofouling traits on reverse osmosis (RO) membranes. We studied the potential activity of RO biofilms at metatranscriptome resolution, accompanied by the morphology and function of the biofouling layer over time, including microscopy and EPS composition, adhesion, and viscoelastic properties. To that end, we cultivated natural multispecies biofilms in RO membranes under treated wastewater flow and extracted RNA to study their taxonomies and gene expression profiles. Concomitantly, the biofilm structure was visualized using both scanning electron microscopy and laser scanning confocal microscopy. We also used quartz crystal microbalance with dissipation to characterize the affinity of EPS to membrane-mimetic sensors and evaluated the viscoelasticity of the Ex-Situ EPS layer formed on the sensor. Our results showed that different active bacterial taxa across five taxonomic classes were assembled on the RO membrane, while the composition shifted between 48 and 96 h. However, regardless of the composition, the maturation of the biofilm resulted in the expression of similar gene families tightly associated with the temporal kinetics of the EPS composition, adhesion, and viscoelasticity. Our findings highlight the temporal selection of specific microbial functions rather than composition, featuring the adhesion kinetics and viscoelastic properties of the RO biofilm.

Effects of plastisphere on phosphorus availability in freshwater system: Critical roles of polymer type and colonizing habitat

Biofilm covered microplastics (BMPs) can act as vectors for the transport of exogenous microbial groups to aquatic ecosystem. However, a consensus regarding the formation and development of BMPs and their effect on phosphorus (P) availability has not been reached. Herein, plastic particles made of fuel-based (PET) and biobased polymers (PLA) were deployed in water and hyporheic zones of an urban river for biofilm colonization. Then, BMPs were transferred to lab incubation to study their effects on the P availability. The results showed that different microplastic biofilms had various bacteria and phytoplankton compositions. Additionally, BMPs induced a shift in the microbial co-occurrence patterns co-differentiated by polymer type and colonizing habitats. Network analyses revealed that the structure of PLA BMPs was more robust, while PET colonized in the hyporheic zone reduced network complexity with looser connections between species, and stronger negatively correlated interactions. However, PET formed denser biofilms by the excretion of extracellular polymeric substances from microalgae, which contributed to the better capacity of P utilization. PET colonized in the water/hyporheic zone significantly decreased soluble reactive phosphate by 42.5 % and 30.8 %, respectively. The abovementioned results indicated that BMPs have the potential to disrupt nutrient availability. This study broadens our perspectives for the ecological effects of BMPs in the aquatic environment.

Evaluation of Antibacterial and Antibiofilm Properties of Kojic Acid against Aeromonas sobria and Staphylococcus saprophyticus

Biofilms composed of microbes and extracellular polymeric substances (EPSs) pose a significant risk to human health and lead to economic loss in the food industry. In this study, the antimicrobial and antibiofilm properties of kojic acid (KA) against Aeromonas sobria (A. sobria) and Staphylococcus saprophyticus (S. saprophyticus) were investigated by determining the leakage of DNA and protein, cell morphology, biofilm formation, the metabolic activity of biofilms, excretion of EPS, and biofilm architecture. The results indicated that the values of minimum inhibitory concentration (MIC) of A. sobria and S. saprophyticus after KA treatment were 0.4 mg/mL and 1.6 mg/mL, respectively. 1 × MIC KA showed unignorable antimicrobial activity against the two bacteria, leading to alterations in the bacterial physicochemical characteristics and cell death. Sub-MICs of KA can inhibit biofilm formation and decrease the metabolic activity and excretion of EPS, and these inhibition effects were in a dose-dependent manner. These results were further confirmed by the visual images obtained from scanning electron microscopy (SEM) and confocal laser scanning microscopy (CLSM). Moreover, S. saprophyticus is more susceptible to KA in inhibiting biofilm formation, and for A. sobria, changes in the cell structure and the permeability of the cell membrane were more obvious. This research highlighted the antibacterial and antibiofilm activity of KA against A. sobria and S. saprophyticus.

Nutrient starvation intensifies chlorine disinfection-stressed biofilm formation

Bacterial surface attachment and subsequent biofilm expansion represent an essential adaptation to environmental signals and stresses, which are of great concern for many natural and engineered ecosystems. Yet the underlying mechanisms and driving forces of biofilm formation in a chlorinated and nutrient-restricted system remain sketchy. In this study, we coupled an experimental investigation and modeling simulation to understand how chlorination and nutrient limitation conspire to form biofilm using Pseudomonas aeruginosa as a model bacterium. Experimental results showed that moderate chlorination at 1.0 mg/L led to biofilm development amplified to 2.6 times of those without chlorine, while additional nutrient limitation (of 1/50-diluted or 0.4 g/L LB broth culture) achieved 4.6 times increment as compared to those of undiluted scenarios (of 20 g/L LB broth culture) with absence of chlorination after 24 h exposure. Meanwhile, intermediate chlorination stimulated instant flagellar motility and subsequently extracellular polymeric substances (EPS) secretion, particularly under limited nutrient condition (of 1/50-diluted or 0.4 g/L LB broth culture) that retarded chlorine consumption and provoked bacterial nutrient-limitation response. From our simulations, chlorine and resource levels along with associated spatio-temporal variations collectively drove bacterial cell movement and EPS excretion. Our results demonstrated that restraining nutrient intensified chlorination-excited cell movement and EPS production that reinforced biological and cell-surface interactions, thereby encouraging bacterial surface attachment and subsequent biofilm development. The findings provide the insights into the linkage of disinfectant and nutrient-regulated bacterial functional responses with consequent micro-habitats and biofilm dynamics.

Effect of stepwise or one-time illumination strategy on the development of algal-bacterial aerobic granular sludge in sequencing batch reactor

Algal-bacterial aerobic granular sludge (AGS) as a promising wastewater treatment biotechnology has attracted more attention in recent years. Still, limited information is available on the effect of stepwise illumination on algal-bacterial AGS. In this study, two illumination strategies, i.e., stepwise (R1) and one-time (R2) daily, were applied to cultivate algal-bacterial AGS from bacterial AGS. Results indicated that R1 holds more advantages over R2 in terms of algal-bacterial AGS formation, biomass growth, extracellular polymeric substances excretion and phosphorus (P) accumulation. Specifically, algal-bacterial AGS in R1 contained higher chlorophyll a content of 5.84 mg/g-MLVSS with the biomass yield of 0.50 g-MLVSS/g-COD in comparison to 5.04 mg/g-MLVSS and 0.43 g-MLVSS/g-COD in R2. Moreover, stepwise illumination effectively alleviated the negative effects of microalgae on anaerobic P release and P accumulation. However, no significant difference was detected between R1 and R2 regarding granule diameter and morphology in addition to organics and nutrients removal.

Phthalate Esters Released from Plastics Promote Biofilm Formation and Chlorine Resistance.

Phthalate esters (PAEs) are commonly released from plastic pipes in some water distribution systems. Here, we show that exposure to a low concentration (1-10 μg/L) of three PAEs (dimethyl phthalate (DMP), di-n-hexyl phthalate (DnHP), and di-(2-ethylhexyl) phthalate (DEHP)) promotes Pseudomonas biofilm formation and resistance to free chlorine. At PAE concentrations ranging from 1 to 5 μg/L, genes coding for quorum sensing, extracellular polymeric substances excretion, and oxidative stress resistance were upregulated by 2.7- to 16.8-fold, 2.1- to 18.9-fold, and 1.6- to 9.9-fold, respectively. Accordingly, more biofilm matrix was produced and the polysaccharide and eDNA contents increased by 30.3-82.3 and 10.3-39.3%, respectively, relative to the unexposed controls. Confocal laser scanning microscopy showed that PAE exposure stimulated biofilm densification (volumetric fraction increased from 27.1 to 38.0-50.6%), which would hinder disinfectant diffusion. Biofilm densification was verified by atomic force microscopy, which measured an increase of elastic modulus by 2.0- to 3.2-fold. PAE exposure also stimulated the antioxidative system, with cell-normalized superoxide dismutase, catalase, and glutathione activities increasing by 1.8- to 3.0-fold, 1.0- to 2.0-fold, and 1.2- to 1.6-fold, respectively. This likely protected cells against oxidative damage by chlorine. Overall, we demonstrate that biofilm exposure to environmentally relevant levels of PAEs can upregulate molecular processes and physiologic changes that promote biofilm densification and antioxidative system expression, which enhance biofilm resistance to disinfectants.

Response of anaerobic granular sludge to long-term loading of roxarsone: From macro- to micro-scale perspective

Extensive use of organoarsenic feed additives such as roxarsone has caused organoarsenicals to occur in livestock wastewater and further within anaerobic wastewater treatment systems. Currently, information on the long-term impacts of roxarsone on anaerobic granular sludge (AGS) activity and the underlying mechanisms is very limited. In this study, the response of AGS to long-term loading of roxarsone was investigated using a laboratory up-flow anaerobic sludge blanket reactor spiked with 5.0 mg L−1 of roxarsone. Under the effect of roxarsone, methane production decreased by ∼40% due to the complete inhibition on acetoclastic methanogenic activity on day 260, before being restored eventually. Over 30% of the influent arsenic was accumulated in the AGS and the capability of AGS to prevent intracellular As(III) accumulation increased with time. The AGS size was reduced by ∼30% to 1.20‒1.26 mm. Based on morphology and confocal laser scanning microscopy analysis, roxarsone exposure stimulated the excretion of extracellular polymeric substances and the surface spalling of AGS. High-throughput sequencing analysis further indicated roxarsone initially altered the acidogenic pathway and severely inhibited the acetoclastic methanogen Methanothrix. Acetogenic bacteria and Methanothrix were finally enriched and became the main contributor for a full restoration of the initial methane production. These findings provide a deeper understanding on the effect of organoarsenicals on AGS, which is highly beneficial for the effective anaerobic treatment of organoarsenic-bearing wastewater.

Extracellular polymeric substances in green alga facilitate microplastic deposition

Microplastics (MPs) are globally ubiquitous in sediments and surface waters. Interactions between biota and MPs are complex and influence their fate and effects in the environment. Once MPs enter aquatic systems, they are colonized by biofilms that may form from the excretion of extracellular polymeric substances (EPS) from microalgae. Biofilm accumulation may change the density of the MPs, contributing to their transport to the sediments. Furthermore, benthic plantivores may consume biofilm laden MPs allowing them to enter the food web. Thus, it is crucial to understand the role algae plays in the vertical transport of MPs in the aquatic environment. In this study, Chlamydomonas was cultured with MPs at different concentrations (0–0.4 mg/mL), and temperatures ranging from 2.5 to 32.5 °C to understand the deposition dynamics and impacts of MPs on EPS production and algal density. Temperatures ranging up to 25 °C increased algal density and MPs deposition. However, at 32.5 °C, algal density and MPs deposition declined. The quantity of MPs also affected algal cell density and EPS production. MPs concentration from 0 to 0.4 mg/mL increased EPS production at all temperatures. Similarly, an increase in algal cell density and MPs deposition occurred when MPs concentration was raised to 0.3 mg/mL. Algal cultures exposed to 0.3–0.4 mg/mL of MPs had a decrease in algal cell density, with no corresponding decline in EPS production. At certain conditions, MPs can facilitate biofilm formation by stimulating EPS production, which can increase cell density thereby expediting MPs transport to the sediment.

Inhibition mitigation of methanogenesis processes by conductive materials: A critical review

Methanogenesis can be promoted by the addition of conductive materials. Although stimulating effects of conductive materials on methane (CH4) production has been extensively reported, the crucial roles on recovering methanogenic activities under inhibitory conditions have not been systematically discussed. This critical review presents the current findings on the effects of conductive materials in methanogenic systems under volatile fatty acids (VFAs), ammonia, sulfate, and nano-cytotoxicity stressed conditions. Conductive materials induce fast VFAs degradation, avoiding VFAs accumulation during anaerobic digestion. Under high ammonia concentrations, conductive materials may ensure sufficient energy conservation for methanogens to maintain intracellular pH and proton balance. When encountering the competition of sulfate-reducing bacteria, conductive materials can benefit electron competitive capability of methanogens, recovering CH4 production activity. Conductive nanomaterials stimulate the excretion of extracellular polymeric substances, which can prevent cells from nano-cytotoxicity. Future perspectives about unraveling mitigation mechanisms induced by conductive materials in methanogenesis processes are further discussed.

Distinction between Cr and other heavy–metal–resistant bacteria involved in C/N cycling in contaminated soils of copper producing sites

For typical copper producing provinces of Heilongjiang, Henan, Inner Mongolia, Jiangxi, Shandong, Tibet, and Yunnan in China, 90 % of sampling sites were heavily polluted with multiple heavy metals. Soil heterogeneity and mutual interference of multimetals are obstacles to explore bacterial resistance pathways in contaminated field soils. Through analyses of contamination indices and bioindicators, combined with multivariate statistical models, the antioxidant enzyme activity, urease–induced precipitation of heavy metals, excretion of extracellular polymeric substances (EPS) were attributed to different types of heavy metals. Furthermore, through redundancy analysis combined with phylogenetic analysis of metal–resistant bacteria, we identified that Verrucomicrobia, Acidobacteria, and Planctomycetes secreted EPS–polysaccharides and EPS–proteins to detoxify Cr, a metal with lower concentrations and lower ecological risk as compared to other metals. The pathway was innovatively differentiated from the multimetal resistance pathways in urease and/or catalase–producing bacteria such as Proteobacteria, Firmicutes, BRC1, Bacteroidetes, Dadabacteria, Entotheonellaeota, Nitrospirae, and Gemmatimonadetes using field studies and high–throughput sequencing. Moreover, these metal–resistant bacteria were linked to C/N cycling processes of urea hydrolysis, nitrification, denitrification, EPS production, and calcite precipitation. It will provide new insight into soil bacterial resistance to multimetals in field studies.

Tetracyclines lead to ammonium accumulation during nitrification process

The effect of tetracyclines used for swine food-production (tetracycline and oxytetracycline) on enriched nitrifying bacteria cultures over time was investigated in this study. Short-term exposure assays were performed in different concentrations of each antibiotic, using ammonia oxidizing bacteria (AOB) culture and nitrifying bacteria. The results pointed out a higher inhibitory effect of tetracycline on both bacterial communities. The AOB was more sensitive to antibiotic exposure when compared to the nitrifying culture. Although high antibiotic concentrations were applied, the half maximal inhibitory concentration (IC50) was achieved only for the AOB culture exposed to tetracycline at a concentration of 273 mg L−1. Nonetheless, the long-term exposure assay demonstrated a reduction of the tetracycline inhibition effect against AOB. The exposure to 100 mg L−1 of tetracycline (TC) did not show relevant influence over ammonium conversion efficiency; however, at 128 mg L−1 of TC, the efficiency decreased from 94% to 72%. Further investigation revealed that TC reduced the final effluent quality due to the development of a resistance mechanism by AOB culture against this antibiotic. This mechanism involves increasing the excretion of extracellular polymeric substances (EPS) and soluble microbial products (SMP), which probably increases BOD, and reduces ammonia consumption by the bacterial culture.

Enhanced excretion of extracellular polymeric substances associated with nonylphenol tolerance in Dictyosphaerium sp

Dictyosphaerium sp. is tolerant to nonylphenol (NP); however, knowledge regarding the mechanisms involved in NP tolerance is limited. In this study, a batch of algal culture experiments were carried out to elucidate the underlying mechanisms by investigating the production and composition of extracellular polymeric substances (EPS) in algae exposed to NP. The excretion of EPS was significantly enhanced (P < 0.001) in algae exposed to 4 and 8 mg/L of NP. The polysaccharides in soluble EPS and the proteins in bound EPS were specifically overproduced. The three-dimensional excitation and emission matrix fluorescence spectra analyses indicated that tyrosine- and tryptophan-like substances were the main functional compositions in the proteins of EPS. In addition, enhanced EPS secretion significantly alleviated the toxicity of NP to the algae by the reduction of cell internalization, as indicated by the higher IC50, biomass, and cell growth rate in the algae with EPS. These discoveries along with the characterizations by algal cell surface hydrophobicity analysis, scanning electron microscopy, and Fourier transform infrared spectra spectroscopy demonstrated the vital role of EPS in the algal resistance to NP.

Cyanobacterial-algal crusts from Late Ediacaran paleosols of the East European Craton

The top surface of the Ediacaran flood basalts of West Ukraine (Volyn), extruded during the break-up of Rodinia supercontinent, is weathered into several meters thick paleosol, covered by late Ediacaran (ca. 550 Ma) sediments. The paleosol grades from unaltered basaltic material into saprock and finally clay-dominated saprolite with a minor content of the organic matter. The upper parts are dominated by kaolinite and hematite. SEM images and SEM-EDS analyses revealed in one of the paleosol profiles a horizon with significant amount of morphologically recognizable organic structures, mineralized by silicates and iron oxides. HF-etching of the bulk rock samples revealed lithic fragments covered irregularly by a biocrust of colonial coccoid cyanobacteria associated with irregularly distributed unicellular algae. The cyanobacteria are preserved as mineralized common mucilage sheaths (glycocalyx), which reproduce the original shape typical of sheaths of modern coccoidal cyanobacterial biofilms, whereas the heavily mineralized algae show features characteristic of modern chlorococcaleans. The preservation of morphologically distinguishable remnants of cyanobacterial biofilms and aggregates of unicellular algae in terrestrial weathering deposits is a rare preservational phenomenon requiring rapid post-mortem permineralization of the dead microbiota. One of the main factors facilitating such a unique fossilization was supposedly a copious excretion of extracellular polymeric substances (EPS) by the biocrust forming biota in which early mineral nucleation occurred. The colonization of the Late Ediacaran land by the presumably desiccation-resistant cyanobacteria-dominated crust communities associated with algae was of importance for creating fertile soils and a soil-forming microbial ecosystem that later allowed a successful land invasion by the ancestors of vascular plants.

Enhanced stable long-term operation of biotrickling filters treating VOCs by low-dose ozonation and its affecting mechanism on biofilm

For long-term operation of highly loaded biotrickling filters (BTFs), the prevention of excess biomass accumulation was essential for avoiding BTF failure. In this study, we proposed low-dose ozonation as a biomass control strategy to maintain high removal efficiencies of volatile organic compounds (VOCs) over extended operation of BTFs. To obtain an optimized biomass control strategy, the relative performance of five parallel BTFs receiving different ozone doses was determined, and the affecting mechanism of ozonation on biofilm was elucidated. Experimental results showed that the decline in ozone-free BTF performance began from day 150, which was correlated with excess biomass accumulation, abundant excretion of extracellular polymeric substances (EPS) and a decline in metabolic activity of biofilm over extended operation. Ozone of 5–10 mg m−3 was effective in preventing excessive growth and uneven distribution of biomass, and eventually maintaining long-term stable operations. Ozone of over 20 mg m−3 possibly inhibited microorganism growth severely, thereby deteriorating the elimination performance instead. Comparison of the biofilm EPS indicated that the presence of ozone reduce EPS contents to different extents, which was possibly beneficial for mass transfer and metabolic activity. Comparative community analysis showed that ozonation resulted in different microbial communities in the BTFs. Dyella was found to be the most abundant bacterial genera in all BTFs regardless of ozonation, indicating strong resistance to ozonation. Chryseobacterium and Burkholderia members were markedly enriched in the ozone-added biofilm, implying good adaptation to ozone presence. These findings provided an improved understanding of low-dose ozonation in maintaining a stable long-term operation of BTF.

Light Regimes Shape Utilization of Extracellular Organic C and N in a Cyanobacterial Biofilm.

ABSTRACTAlthough it is becoming clear that many microbial primary producers can also play a role as organic consumers, we know very little about the metabolic regulation of photoautotroph organic matter consumption. Cyanobacteria in phototrophic biofilms can reuse extracellular organic carbon, but the metabolic drivers of extracellular processes are surprisingly complex. We investigated the metabolic foundations of organic matter reuse by comparing exoproteome composition and incorporation of 13C-labeled and 15N-labeled cyanobacterial extracellular organic matter (EOM) in a unicyanobacterial biofilm incubated using different light regimes. In the light and the dark, cyanobacterial direct organic C assimilation accounted for 32% and 43%, respectively, of all organic C assimilation in the community. Under photosynthesis conditions, we measured increased excretion of extracellular polymeric substances (EPS) and proteins involved in micronutrient transport, suggesting that requirements for micronutrients may drive EOM assimilation during daylight hours. This interpretation was supported by photosynthesis inhibition experiments, in which cyanobacteria incorporated N-rich EOM-derived material. In contrast, under dark, C-starved conditions, cyanobacteria incorporated C-rich EOM-derived organic matter, decreased excretion of EPS, and showed an increased abundance of degradative exoproteins, demonstrating the use of the extracellular domain for C storage. Sequence-structure modeling of one of these exoproteins predicted a specific hydrolytic activity that was subsequently detected, confirming increased EOM degradation in the dark. Associated heterotrophic bacteria increased in abundance and upregulated transport proteins under dark relative to light conditions. Taken together, our results indicate that biofilm cyanobacteria are successful competitors for organic C and N and that cyanobacterial nutrient and energy requirements control the use of EOM.

Effect of TiO2 nanoparticles on aerobic granulation of algal–bacterial symbiosis system and nutrients removal from synthetic wastewater

The influence of TiO2 nanoparticles (TiO2-NPs) (10–50mg/L) on aerobic granulation of algal–bacterial symbiosis system was investigated by using two identical sequencing batch reactors (SBRs). Although little adverse effect was observed on their nitritation efficiency (98–100% in both reactors), algal–bacterial granules in the control SBR (Rc) gradually lost stability mainly brought about by algae growth. TiO2-NPs addition to RT was found to enhance the granulation process achieving stable and compact algal–bacterial granules with remarkably improved nitratation thus little nitrite accumulation in RT when influent TiO2-NPs⩾30mg/L. Despite almost similar organics and phosphorus removals obtained in both reactors, the stably high nitratation efficiency in addition to much stable granular structure in RT suggests that TiO2-NPs addition might be a promising remedy for the long-term operation of algal–bacterial granular system, most probably attributable to the stimulated excretion of extracellular polymeric substances and less filamentous TM7.

Long-term effect of the antibiotic cefalexin on methane production during waste activated sludge anaerobic digestion

Long-term experiments herein were conducted to investigate the effect of cefalexin (CLX) on methane production during waste activated sludge (WAS) anaerobic digestion. CLX exhibited a considerable inhibition in methane production during the initial 25days while the negative effect attenuated subsequently and methane production recovered depending on CLX doses used (600 and 1000mg/L). The highest methane yield reached 450mL at 1000mg-CLX/L after 157days of digestion, 63.8% higher than CLX-free one. Stimulated excretion of extracellular polymeric substances (EPS) by CLX served as microbial protecting layers, creating a suitable environment for microbes’ growth and fermentation. Further examination via ultraviolet visible (UV–Vis) spectra also verified the elevated slime EPS, LB-EPS and TB-EPS indicated by UV-254 in the presence of CLX. Unlike the commonly accepted adverse effect, this study demonstrated the beneficial role of CLX in methane production, providing new insights into its true environmental impacts.

Evolution of bioaggregate strength during aerobic granular sludge formation

This work investigated the modification of aggregate properties during the formation of granular sludge in a sequencing batch airlift reactor (SBAR). The cohesion of biological aggregates was quantified by subjecting sludge samples to two different controlled shear stresses in a stirred reactor. For reference sludge (without granules), flocs broke and reformed easily, indicating that floc size was controlled by the turbulence micro-scale (Kolmogorov scale, here from 17 μm to 62 μm). In contrast, granules showed high strength which enabled them to resist turbulence and their size was no longer imposed by the Kolmogorov micro-scale. Different steps were observed during the granulation process: a first increase of aggregate cohesion associated with a decrease in sludge volume index (SVI), a growth of aggregates with detachment of fragile particles from the surface and, finally, an increase in the sizes of small and large granules to reach a pseudo-stable size distribution. Results suggest that small particles could have formed the seeds for new granules, as they were maintained in the bioreactor. Here, granular sludge was formed in an SBAR with a conventional settling time (30 min), i.e. without particle washout, and with a low superficial air velocity (SAV = 0.6 cm s −1): it is thus demonstrated that high SAV and low settling time are not necessary to produce granules, but probably only accelerate the accumulation of granules. It is shown that the increase of cohesion is the initial phenomenon explaining the granule formation concomitantly with bacterial aggregates densification. It seems important, in the future, to investigate the reasons for this cohesion increase, which is possibly explained either by bacterial bounding interactions or the excretion of extracellular polymeric substances (EPS).

Acoustic sensing of the bacterium–substratum interface using QCM-D and the influence of extracellular polymeric substances

It is commonly assumed that bacterial presence on a QCM sensor-surface is associated with a negative frequency shift according to conventional mass-loading theory. Here, we demonstrate that bacteria adhering to QCM sensor-surface may yield positive frequency shifts up to 1.9 × 10 −6 Hz per bacterium according to a coupled-oscillator theory. Furthermore, it is demonstrated that the excretion of extracellular polymeric substances (EPS) by adhering bacteria can change the frequency shift in the negative direction by 1.7 × 10 −6 Hz per bacterium, according to conventional mass-loading theory. The difference in frequency shifts between an EPS-producing and a non-EPS producing staphylococcal strain correlated with the excretion of 3 × 10 −14 g EPS per bacterium, representing only a few percent of the weight of a bacterium. Thus an adsorbed molecular mass as low as a few percent of the mass of an adhering bacterium significantly alters the QCM-signal. Since adhesion of many different bacterial strains is accompanied by molecular adsorption of EPS, with potentially opposite effects on the QCM-signal, a combination of the coupled-oscillator and normal mass-loading theory has to be applied for proper interpretation of QCM-frequency shifts in bacterial detection.

Physical characterization and diel dynamics of different fractions of extracellular polysaccharides in an axenic culture of a benthic diatom

The excretion of extracellular polymeric substances (EPS) by an axenic culture of the benthic diatom Cylindrotheca closterium was investigated. Two sequential extraction steps proved to be suficient to remove the bulk of the EPS present. Soluble EPS was recovered by a simple centrifugation step and represented a fraction that was not or was only loosely associated with diatom cells. For the extraction of bound EPS, different procedures were compared. The best results were obtained using distilled water as extraction solvent (1 h, 30 °C). The sugars that were recovered using this procedure were typically associated with aggregates of diatoms. In addition to the distinct differences in localization of the different types of EPS, their temporal dynamics differed in relation to the light–dark cycle. Soluble EPS were continuously released into the medium at a rate of 1.6 pg cell−1 day−1. In contrast, the production of bound EPS was highly light-dependent. In the dark, this bound EPS rapidly disappeared, probably as the result of its utilization by the diatoms.