Presence Of Extracellular Polymeric Substances Research Articles

Immobilised antimicrobial peptides in downregulation of biofilm.

Colonisation of sessile bacterial species on biotic and abiotic surfaces is responsible for the development of various infections in humans. At present, biofilm-associated chronic infections have been a prime concern among the healthcare practitioners since they are impermeable to drugs, resulting in the development of antibiotic resistance or multi-drug resistance. For a few decades, a lot of research activity has been performed in the development of alternative therapeutics to combat biofilm-associated chronic infections. The presence of extracellular polymeric substance (EPS) prevents the permeation of most of the drugs rendering drug failures. The use of small molecules has been necessary to penetrate easily through the EPS and act on the targeted cells. In present days, the use of antimicrobial peptides (AMPs) has gained immense importance as alternative therapeutics since they exhibit a novel class of antibiotics exhibiting a wide spectrum of activity and possess a low rate of development of resistance. In the last few decades, a large number of AMPs have been identified from varied groups of organisms as effector molecules for innate immune system acting as an important line of defence. In this review, we will discuss the use of AMPs as effective agents to combat various biofilm-associated chronic infections.

Dissolved Mn2+ promotes microbially-catalyzed protodolomite precipitation in brackish oxidized water

Manganese (Mn) is a common trace element in dolomite, yet its precise role in dolomite precipitation, if any, has not been explored. This study aims to investigate carbonate mineral formation in the presence of extracellular polymeric substances produced by Bacillus licheniformis Y1 in 14 cross-over experiments within the MgCl2-CaCl2-MnCl2 system. The study involved the analysis and comparison of water chemistry, mineralogy and bacterial metabolites. The findings indicate that B. licheniformis Y1 induces the formation of Mg-free calcite and low-Mg calcite in Mn-free solutions (Mg/Ca < 2). The addition of 0.001 mol/L Mn2+ results in the precipitation of Mg-rich calcite. In the presence of a relatively high Mg2+ content in the solution (Mg/Ca > 6), B. licheniformis Y1 in a Mn-free solution induces the formation of aragonite and dypingite, whereas the addition of 0.001 mol/L Mn2+ leads to the formation of Mn-bearing dolomite in the precipitate. Notably, at Mg/Ca = 6, an increase in Mn2+ content of the fluid to 0.002 and 0.003 mol/L yields Mn-rich dolomite. The Mn-bearing dolomite exhibits a regular spherical aggregate morphology with diameters ranging from 15 to 60 μm. The 2θ position of the (104) crystal face is close to that of dolomite, and the thermal decomposition temperature is lower than that of calcite. XPS results show that MnO bonds exist on the mineral surface. Acidic amino acids such as glutamic acid as well as humic acid in EPS, especially fulvic acid analogues, actually rise with increasing Mn2+ concentration and contain a large number of functional groups including carboxyl groups. Molecular dynamics simulations show that fulvic acid can effectively promote the dehydration of Mg-water complexes, thus promoting the formation of dolomite. These experiments indicate that the presence of Mn ions in the solution, along with bacteria and EPS, promote the precipitation of protodolomite.

Fabric Fiber as a Biofilm Carrier for Halomonas sp. H09 Mixed with Lactobacillus rhamnosus GG.

Biofilm bacteria have stronger resistance to the adverse external environment compared to planktonic bacteria, and biofilms of non-pathogenic bacteria have strong potential for applications in food. In this experiment, Halomonas sp. H09 and Lactobacillus rhamnosus GG, which have film-forming ability in monoculture and better film-forming ability in mixed culture than the two strains alone, were selected as the target strains for mixed culture. According to SEM observation and bacterial dry weight measurement, the target strain formed a dense biofilm on a 0.1g/L chitosan-modified cellulose III carrier. Furthermore, the presence of extracellular polymeric substances in biofilms was verified by EDS and FTIR. The results showed that 0.1g/L chitosan-modified cellulose III was an ideal carrier material for immobilization of Halomonas sp. H09 with Lactobacillus rhamnosus GG biofilm. This research provided a basis for the selection of non-pathogenic mixed-bacteria biofilm carriers.

Investigation and prediction of the biotoxicity of Cu2+ to Chlorella vulgaris: modification of the biotic ligand model.

The increased copper ion (Cu2+) concentrations in aquatic ecosystem significantly influence the environmental quality and ecosystem safety, while information on the Cu2+ biotoxicity to aquatic microorganisms and the models for biotoxicity prediction are still unclear. In this study, the toxicities of Cu2+ to Chlorella vulgaris under different environmental conditions (e.g., Na+, K+, Ca2+, Mg2+, pH, and dissolved organic matter) were explored, with the experimental results in comparison with those predicted by the biotic ligand model (BLM). Results showed that increased Cu2+ concentration caused obvious toxicities to C. vulgaris, whereas the commonly occurring cations and dissolved organic matters can protect the metabolism system of C. vulgaris. The presence of extracellular polymeric substances (EPS) matrix can alleviate the biotoxicity via increasing the surface biosorption but decreasing cell internalization of Cu2+ in C. vulgaris. Due to the presence of EPS matrix, the experimental biotoxicity results under each condition were significantly lower than those predicted by the BLM model, which was thus modified via taking the EPS matrix as the supplement of allochthonous organic matters. After that, the modified BLM was characterized with a higher degree of precision and can be used in natural waters for biotoxicity prediction. Results obtained can enhance our insights into the ecological effects and biotoxicity prediction of heavy metals in natural aquatic ecosystems.

Production and composition of extracellular polymeric substances by a unicellular strain and natural colonies of Microcystis: Impact of salinity and nutrient stress.

The transfer of toxic cyanobacterial Microcystis blooms from freshwater to estuaries constitutes a serious environmental problem worldwide that is expected to expand in scale and intensity with anthropogenic and climate change. The formation and maintenance of Microcystis in colonial form is conditioned to the presence of extracellular polymeric substances (EPS). In this study, we attempted to better understand how the mucilaginous colonial form of Microcystis evolves under environmental stress conditions. In particular, we studied and compared the production and the composition of EPS fractions (attached and free) from natural colonies of a Microcystis bloom and from a unicellular M. aeruginosa strain under salinity and nutrient stress (representing a land-sea continuum). Our results highlighted a greater production of EPS from the natural colonies of Microcystis than the unicellular one under nutrient and combined stress conditions dominated by the attached form. In comparison to the unicellular Microcystis, EPS produced by the colonial form were characterized by high molecular weight polysaccharides which were enriched in uronic acids and hexosamines, notably for the free fraction in response to increased salinities. This complex extracellular matrix gives the cells the ability to aggregate and allows the colonial cyanobacterial population to cope with osmotic shock.

Microbially influenced formation of anhydrite at low temperature

Calcium sulfate minerals are abundant in nature – on Earth and on Mars – and important in several fields of material sciences. With respect to gypsum and bassanite, anhydrite represents the anhydrous crystalline phase in the CaSO4–H2O system. Despite years of research, the formation of anhydrite in the laboratory at low temperature remains challenging and, in the geological record, this mineral is mostly interpreted as a secondary phase that form through metamorphic dehydration of gypsum. Here, we present the results of laboratory precipitation experiments showing that anhydrite can form at 35 °C from evaporated seawater through a microbially influenced mineralization process. The experiments were conducted in the presence of extracellular polymeric substances (EPS) produced by bacterial strains isolated from a modern evaporitic environment, the Dohat Faishakh sabkha in Qatar. Without organic molecules, only gypsum formed in parallel control experiments. This finding provides a possible explanation for the origin of several natural occurrences of anhydrite that cannot be satisfactorily explained by existing models and reveals a new precipitation pathway that may have industrial applications.

Roles of extracellular polymeric substances on Microcystis aeruginosa exposed to different sizes of polystyrene microplastics

Extracellular polymeric substances (EPS) are important shields for microalgae when confronting with external stresses. However, the underlying roles of EPS in the interactions between microplastics (MPs) and microalgae remain poorly understood. In this study, three sizes of polystyrene (PS) MPs (20 nm, 100 nm, and 1 μm) were chosen for evaluating the compositions of EPS, secreted by Microcystis aeruginosa during exposure. The results indicated that the EPS compositions were different when M. aeruginosa was exposed to PS MPs of different sizes. The presence of EPS is helpful for alleviating the adverse effects of PS MPs on M. aeruginosa cell growth, photosynthesis, and oxidative stress. With the exception of the shading effect, insufficient EPS cause direct adsorption of unstable 1 μm PS MPs to the algal surface, which could destroy the cell wall. In contrast, aromatic proteins and fulvic acids are representative EPS components stimulated by 100 nm PS MPs, contributing to the self-aggregation and encapsulation of algal cells and availability of nutrients for algal growth, respectively. High amounts of polysaccharides were secreted by M. aeruginosa along with humic acids during exposure to 20 nm PS MPs, both of which are crucial in the homo-aggregation of 20 nm PS MPs toward minimize its adverse effects on M. aeruginosa. Together, these findings revealed the differences in EPS under the stimulation of PS MPs of different sizes and clarified the roles of different EPS components in resisting the adverse effects of PS MPs on M. aeruginosa.

Exploring the role of extracellular polymeric substances in the antimony leaching of tailings by Acidithiobacillus ferrooxidans.

The concentration of Sb bearing tailings in water located in abandoned antimony mines was found to be a big problem, as they contaminate other water resources and entire food chain. Microorganisms were found to be key in tailing leaching and reaction speeding in the presence of extracellular polymeric substances (EPS) produced by bacteria. Herein, we investigated the pattern of the Sb leaching from Sb bearing tailings using Acidithiobacillus ferrooxidans, and analyzed the mechanism of EPS in the leaching process of Sb. To completely and deeply understand the functions of EPS in the bioleaching of antimony tailings, the generation behavior of EPS produced by Acidithiobacillus ferrooxidans (A. ferrooxidans) during bioleaching was characterized by three-dimensional excitation-emission matrix (3D-EEM). Meanwhile, X-ray photoelectron spectroscopy (XPS) and Fourier transform infrared (FT-IR) were used to show the changes of EPS functional groups before and after leaching. Compared with the functional groups in EPS produced by A. ferrooxidans before leaching, the content of hydroxyl and amino groups that reduce high-valent metals to low-valent metals in EPS decreases after leaching, and the carbonyl content increases, corresponding to the ratio of trivalent antimony increased, indicating that EPS could reduce the risk of pentavalent antimony to trivalent one. At the same time, with biological scanning electron microscopy and energy spectrum scanning, the observation of EPS on the mineral surface showed that Sb was adsorbed in the EPS, and the XPS of Sb was fine. Spectral analysis showed that the Sb adsorbed in EPS contained both Sb(III) and Sb(V). Besides, for revealing the influence of EPS in the leaching process of Sb from tailings, this work provided an in-depth understanding of the mechanism of Sb released from tailings under the action of A. ferrooxidans and further provides a basis for the biogeochemical cycle of Sb.

Determination of Compressibility and Relaxation Behavior of Yeast Cell Sediments by Analytical Centrifugation and Comparison with Deposit Formation on Membrane Surfaces.

Separation of cells from produced biomolecules is a challenging task in many biotechnological downstream operations due to deposit formation of the retained cells, affecting permeation of the target product. Compression and relaxation behavior of cell deposits formed during filtration are important factors affecting operational performance. The determination of these factors by flux or pressure stepping experiments is time- and labor-intensive. In this work, we propose a screening method by analytical centrifugation, which is capable of detecting small differences in compression and relaxation behavior induced by milieu changes, using a model system comprised of washed and unwashed yeast cells in the presence or absence of bovine serum albumin as a model target protein. The main effects observed were firstly the impact of pH value, affecting interaction of bovine serum albumin and yeast cells especially close to the isoelectric point, and secondly the effect of washing the yeast cells prior to analysis, where the presence of extracellular polymeric substances led to higher compressibility of the deposited cells. By comparing and validating the obtained results with dead-end filtration trials, the stabilizing role of bovine serum albumin in deposits formed at low pH values due to interactions with the yeast cells was underlined.

Magnesium isotope fractionation during carbonate precipitation associated with bacteria and extracellular polymeric substances

Magnesium (Mg) is abundant on Earth's surface aquatic environments and plays an important role during the precipitation of biogenic CaCO3 minerals. Moreover, Mg isotopes are increasingly used to study a number of bio-induced mineralization processes of calcite. In this study, Mg isotope signatures during calcite and aragonite precipitation in the presence of Curvibacter sp. HJ-1 and of their extracellular polymeric substances (EPS) were investigated and the evolution of mineralogical and liquid composition were tracked over time. The results showed that light Mg isotopes were preferentially incorporated into the precipitated solids in all the experiments with and without bacteria. Significant Mg isotope fractionation was associated with the transformation of amorphous calcium carbonate (ACC) to crystal carbonate (Δ26Mgcrystals-ACC = −0.9‰ and −0.7‰ in biotic and EPS experiments, respectively), and obvious Mg isotope fractionation was noted between solids and liquid (Δ26Mgsolid-liquid achieved −2.2‰ and −2.0‰ in biotic and EPS experiments, respectively). Besides, δ26Mg values were significantly correlated with pH, Mg content in liquid and solid, bacterial density and presence of EPS in biotic experiments. These findings indicated that strain HJ-1 had an effect on Mg isotope fractionation during calcite and aragonite precipitation. The difference in Mg isotope fractionation may be a new tool for understanding the biologically mediated effects on Mg-bearing carbonate precipitation, and serves as a useful alternative for aqueous Mg isotope, which is vital for reconstructing past environmental changes.

Enriched sewage sludge from anaerobic pre-treatment in spurring valorization potential of black soldier fly larvae

The valorization of sewage sludge by black soldier fly larvae (BSFL) has gained attentions for sewage sludge management since the sludge can be reduced securely as well as larval biomass can be used for biorefineries application. Nevertheless, the BSFL growth was impeded while assimilating nutrition from sewage sludge due to the presence of extracellular polymeric substances (EPS) that had entrapped the essential nutrients inside. Accordingly, the pre-treatment of sewage sludge via anaerobic digestion at different pH was employed in this work to rupture the EPS structure and release more nutrients for larval growth. The results showed that larvae fed with raw sewage sludge had attained the lowest final larval weight (2.05 ± 0.38 mg/larva) as opposed to batches fed with pre-treated sewage sludges. This was because the soluble carbohydrate (more than 6.81 ± 1.31 mg of glucose/g sewage sludge) in EPS was released after anaerobic pre-treatment, facilitating larval assimilation for growth. Furthermore, it was observed that further increasing of pH for sewage sludge pre-treatment had led to lower final larval weight gained due to the inhibitory effect stemming from ammonia production at higher pH. The anaerobic pre-treatment of sewage sludge being executed at pH 3 for 8 days had achieved the highest final larval weight at 7.34 ± 0.97 mg/larva. The still low quality of sewage sludges after the pre-treatment also offered benefit, where high sewage sludge reduction and waste reduction index were recorded due to the necessity of BSFL to consume more sewage sludge in compensating the nutrients destitution in sludge. Lastly, the possibility of predicting final larval weight was successfully materialized via a statistical model derived from the multiple linear regression method. The derived model incorporated the interactive parameters of anaerobic pre-treated pH and durations at various combinations could predict the final larval weight.

Production of Hydroxyl Radicals from Oxygenation of Simulated Acid Mine Drainage in Presence of Extracellular Polymers Substances.

The reaction mechanisms Fe(II) abiotic oxidation produce ·OH by CaCO3-induced in acid mine drainage (AMD) are well-documented, but little is known about the influence of extracellular polymeric substances (EPS) secreted by microorganisms on Fe(II) oxidation in AMD. In this study, ·OH production was experimently measured from oxygenation of simulated AMD in the presence of EPS. The cumulative ·OH increased from 56.75 to 158.70 µM within 24h at pH 3 with the increase in EPS concentration from 0 to 12mg/L. An appropriate pH (about 6) and EPS (6mg/L) concentration were required for the moderate rate of Fe(II) oxidation. Besides, the yield of ·OH increased remarkably with the addition of Fe3+. In the presence of EPS, ·OH production is attributed mainly the complexation of Fe(II) with EPS, of which is rich of carboxyl and hydroxyl groups. The findings provide fundamental supplement of ·OH production from Fe(II) oxidation by microorganisms in natural AMD.

Correlating black soldier fly larvae growths with soluble nutrients derived from thermally pre-treated waste activated sludge

Black soldier fly larvae (BSFL) have been deployed to valorize various organic wastes. Nonetheless, its growth rate whilst being offered with waste activated sludge (WAS) is not promising, likely by virtue of the presence of extracellular polymeric substances’ structure in WAS. In this work, the WAS were first thermally pre-treated under different treatment temperatures and durations before being administered as the feeding substrates for BSFL. The results showed the thermal pre-treatment could improve WAS palatability and subsequently, enhance the growth of BSFL especially after the pre-treatments at 75 °C and above. The highest larva weight gained was recorded at 2.16 mg/larva for the WAS sample being pre-treated at 90 °C and 16 h. Furthermore, the samples pre-treated above 75 °C also achieved higher degradation rates, indicating that the 75 °C was a threshold temperature to effectively hydrolyze the WAS. The changes of WAS characteristics, namely, (i) soluble chemical oxygen demand (SCOD), (ii) soluble carbohydrate, (iii) soluble protein, (iv) humic substances and (v) total soluble protein and humic substances, after the thermal pre-treatments were also studied in correlating with the BSFL growth. Accordingly, a model was successfully developed with the highest R2 value attained at 0.95, evidencing the SCOD was the most suitable WAS characteristic to accurately predict the BSFL growth behavior.

Brevundimonas diminuta isolated from mines polluted soil immobilized cadmium (Cd2+) and zinc (Zn2+) through calcium carbonate precipitation: Microscopic and spectroscopic investigations

The toxic metal(loid)s TMs resistant bacterium Brevundimonas diminuta was isolated for the first time from mines polluted soil in Fengxian, China, and assessed for its potential for Cd and Zn precipitation in Cd and Zn co-contaminated aqueous solution at various Cd and Zn levels (20, 40, 80, 160, and 200 mg L−1), pH values (5, 6, 7, 8, and 9), and temperatures (20, 25, 30, and 35 °C). B. diminuta showed a high resistance to both Cd and Zn and was able to precipitate up to 99.2 and 99.7% of dissolved Cd and Zn respectively, at a pH of 7 and temperature of 30 °C. B. diminuta reduced the dissolved concentrations of Cd and Zn below the threshold levels in water. The 3D-EEM analysis revealed the presence of extracellular polymeric substances (EPS) such as tryptophan indicating bacterial growth under Cd/Zn stress. FTIR showed polysaccharides, CO32−, CaCO3, PO43−, and proteins, which may enhance bacterial growth and metal precipitation. SEM-EDS confirmed the leaf-like and granular shape of the biological precipitation and reduction in the percent weight of TMs, which promoted the adhesion/adsorption of Cd2+, Zn2+, and Ca2+. Moreover, XRD analysis confirmed the precipitation of Cd, Zn, and Ca in the form of CdCO3/Cd3(PO4)2, ZnCO3/ZnHPO4/Zn2(OH)PO4/Zn3(PO4)2, and CaCO3/Ca5(PO3)4OH, respectively. These findings indicate that Brevundimonas diminuta can be used for the bioremediation of TMs-contaminated aquatic environments.

RETRACTED: Synergism between goethite size and extracellular polymeric substances (EPS) in the formation of mineral–mineral and organo-mineral complexes of soil microaggregates

Soil microaggregates composed of mineral–mineral and organo-mineral complexes are crucial for controlling many soil ecosystem functions such as soil stability and persistence of organic matter. However, mechanisms of how microbial-derived organic matter (e.g., extracellular polymeric substances, EPS) and particle size of the mineral shape mineral–mineral and organo-mineral complexes in multiphase system are unclear. Here, batch experiments (i.e., mineral mixtures of quartz, kaolinite, and three different sizes of goethite) were used to determine the effect of goethite size and EPS on the formation of mineral–mineral and organo-mineral complexes. Our results revealed that the multiphase system containing micro-sized goethite had a higher proportion of 50–250 μm aggregates than these containing nano-sized goethite in the presence of EPS. When EPS was absent, the particle distribution was not related to goethite size, and the mineral suspension was homogeneous. The mechanisms of mineral–mineral and organo-mineral complexes formation were further investigated by measuring zeta potential. Negatively charged EPS adsorbed onto positively charged goethite surface through electrostatic interactions, resulting in stable organo-mineral complexes and inhibition of mineral–mineral complexes. EPS-goethite interaction resulted in a marked decrease of phosphorus in the suspension, suggesting preferred adsorption of EPS-P compounds to goethite. This interaction led to EPS fractionation which was affected by goethite size due to the particle size-dependent surface interactions. The information obtained in this study is of fundamental significance for understanding the initial formation of microaggregates in the edaphic microenvironment.

Influence of extracellular polymeric substances on the heteroaggregation between CeO2 nanoparticles and soil mineral particles

Interaction with soil mineral particles (SMPs) and organic matters can significantly determine the fate of nanoparticles (NPs) in the environment such as waters, sediments, and soils. In this study, the heteroaggregation of CeO2 NPs with different soil minerals (kaolinite, montmorillonite, goethite and hematite) and the influence of extracellular polymeric substance (EPS) were studied. The obvious heteroaggregation between CeO2 NPs with different SMPs were demonstrated via co-settling and aggregation kinetics experiments. The variety in the heteroaggregation between CeO2 NPs with different SMPs is mainly induced by the difference in their surface properties, such as surface charge, specific surface areas and surface complexation. The presence of EPS can result in great inhibition on the heteroaggregation between CeO2 NPs with the positive charged goethite by enhancing the electrostatic repulsion between NPs and mineral colloids. However, the influence of EPS on the interaction between CeO2 NPs with negative charged SMPs is more dependent on the steric stabilization. The presence of EPS may promote the migration of CeO2 NPs in environment and then increase their risks to human health and ecosystems. These findings contribute to better understanding interactions between NPs and SMPs and have important implications on predicting the behaviors and risks of NPs in the natural environment.

Insight into the role of extracellular polymeric substances in denitrifying biofilms under nitrobenzene exposure

Denitrifying biofilm promises to be very useful for remediation of nitro-aromatic compounds (NACs) and nitrates in wastewater. Little is known about the role of extracellular polymeric substances (EPS) in nitrobenzene (NB, a typical NAC) remediation, despite the indispensability of EPS for biofilm formation. Herein, the significance of the mechanistic role of EPS in the response of denitrifying biofilms to various levels of NB was investigated. The removal of NB was predominantly controlled via absorption, with little biodegradation during the short-term exposure. Specifically, NB was adsorbed by EPS, as shown by a total adsorption of 40.06% at the initial step, which declined to around 10.52% in the equilibrium stage, while sorption via cells gradually increased from 59.93% to 89.47% over the same period. The results suggested that EPS might act as an important reservoir for NB, which endows inner cells with increased adsorption ability. The presence of EPS might also alleviate the negative impacts of NB toxicity on inner cells, thus protecting microorganisms. This was indicated by the difference in denitrification performance and cell integrity between intact and EPS-free biofilms. High-throughput sequencing data demonstrated that EPS could maintain the stability of microbial communities under NB stress. The fluorescence quenching analysis further indicated that EPS formed stable complexes with NB mainly through hydrophobic interactions with protein-like fractions (tryptophan and tyrosine). Moreover, Fourier transform infrared spectroscopy identified that the hydroxyl, amino, carboxyl, and phosphate groups of EPS were the candidate functional groups binding with NB. Protein secondary structures were also significantly affected, resulting in a loose structure and enhanced hydrophobic performance for EPS. These results provide insights into the role of EPS in alleviating NB-caused cellular stress and the underlying binding mechanisms between NB and EPS.

Metal pollutant pathways in cohesive coastal catchments: Influence of flocculation and biopolymers on partitioning and flux

The impacts of the partitioning of potentially toxic metals (PTM) within the estuarine environment is highly complex, but is of key significance owing to increases in populations living within such sensitive environments. Although empirical data exist for the partitioning of metals between the dissolved and particulate phases, little is known regarding the impacts of extracellular polymeric substances (EPS) upon the flocculation of particles within such a dynamic system nor the resultant influence on the distribution of metals between the particulate and dissolved phases. This prevents regulators from fully understanding the fate and risks associated with metals in estuaries. This study provides data associated with the simulation of 3 settlings typical of the turbulent mixing found in estuaries and partitioning of copper, cadmium, nickel, arsenic, lead and zinc for 3 salinities (0, 15, 30 PSU) reflecting the full salinity range from freshwater to seawater. Experiments were completed with and without the presence of EPS, using kaolin as the mineral particulate. The results showed significant differences between salinity, PTMs and turbulence for the experiments with and without EPS present. Overall, salinity was the main factor controlling the PTM partitioning to sediment, however the flocculation process did impact on the PTM distribution and with the addition of EPS the impact was more pronounced. The data highlighted the importance of taking account of EPS within any estuarine sediment process modelling, for relying on simple partitioning with corrections for salinity would likely lead to significant bias.

Transport and Retention of Free-Living Amoeba Spores in Porous Media: Effects of Operational Parameters and Extracellular Polymeric Substances.

Amoebas are protists that are widespread in water and soil environments. Some species are pathogenic, inducing potentially lethal effects on humans, making them a major threat to public health. Nonpathogenic amoebas are also of concern because they have the potential to carry a mini-microbiome of bacteria, either transiently or via more long-term stable transport. Due to their resistance to disinfection processes, the physical removal of amoeba by filtration is necessary to prevent their propagation throughout drinking water distribution networks and occurrence in tap water. In this study, a model amoeba species Dictyostelium discoideum was used to study the transport and retention behavior of amoeba spores in porous media. The key factors affecting the transport behavior of amoeba spores in fully saturated media were comprehensively evaluated, with experiments performed using a quartz crystal microbalance with dissipation monitoring (QCM-D) and parallel plate chamber system. The effects of ionic strength (IS) on the deposition of spores were found to be in contrast to the predicted Derjaguin-Landau-Verwey-Overbeek (DLVO) theory that more deposition is observed under lower-IS conditions. The presence of extracellular polymeric substances (EPS) was found to be the main contributor to deposition behavior. Overall, these results provide plausible evidence for the presence of amoeba in tap water. Furthermore, this is one of the first studies to examine the mechanisms affecting the fate of amoeba spores in porous media, providing a significant baseline for future research to minimize the safety risk presented by amoeba in drinking water systems.

Extracellular Polymeric Substances Drive Symbiotic Interactions in Bacterial‒Microalgal Consortia

The importance of several factors that drive the symbiotic interactions between bacteria and microalgae in consortia has been well realised. However, the implication of extracellular polymeric substances (EPS) released by the partners remains unclear. Therefore, the present study focused on the influence of EPS in developing consortia of a bacterium, Variovorax paradoxus IS1, with a microalga, Tetradesmus obliquus IS2 or Coelastrella sp. IS3, all isolated from poultry slaughterhouse wastewater. The bacterium increased the specific growth rates of microalgal species significantly in the consortia by enhancing the uptake of nitrate (88‒99%) and phosphate (92‒95%) besides accumulating higher amounts of carbohydrates and proteins. The EPS obtained from exudates, collected from the bacterial or microalgal cultures, contained numerous phytohormones, vitamins, polysaccharides and amino acids that are likely involved in interspecies interactions. The addition of EPS obtained from V. paradoxus IS1 to the culture medium doubled the growth of both the microalgal strains. The EPS collected from T. obliquus IS2 significantly increased the growth of V. paradoxus IS1, but there was no apparent change in bacterial growth when it was cultured in the presence of EPS from Coelastrella sp. IS3. These observations indicate that the interaction between V. paradoxus IS1 and T. obliquus IS2 was mutualism, while commensalism was the interaction between the bacterial strain and Coelastrella sp. IS3. Our present findings thus, for the first time, unveil the EPS-induced symbiotic interactions among the partners involved in bacterial‒microalgal consortia.