Extracellular Polysaccharide Content Research Articles

Influence of Biofilm Maturity on the Antibacterial Efficacy of Cold Atmospheric Plasma in Oral Microcosm Biofilms.

As biofilms mature, biomass and extracellular polysaccharide (EPS) content increases, enhancing pathogenicity. Therefore, this study aimed to evaluate the antibacterial efficacy of cold atmospheric plasma (CAP) against oral microcosm biofilms and the influence of biofilm maturity on treatment. Oral microcosm biofilms were cultured on hydroxyapatite disks for 2 and 6 days. Based on the treatment and biofilm maturity, these were subsequently allocated into six groups (N = 19 each): Groups 1 and 2 were incubated with distilled water for 1 min; Groups 3 and 4 were treated with CAP for 2 min, and Groups 5 and 6 were treated with 0.12% chlorhexidine gluconate for 1 min. Groups 1, 3, and 5 represent 2-day biofilms, and Groups 2, 4, and 6 represent 6-day biofilms. Treatments were repeated daily for 5 days. Antibacterial efficacy was analyzed by measuring oral biofilms' red fluorescence intensity (RatioR/G) and quantifying EPS content and bacterial viability. The RatioR/G was 1.089-fold and 1.104-fold higher in Groups 4 and 6 than in Groups 3 and 5 following antibacterial treatment, respectively (p < 0.001). EPS content increased by 1.71-fold in Group 6 than in Group 5 (p < 0.001). Bacterial survival rate was the lowest in Group 3 (p = 0.005). These findings underscore the relevance of CAP treatment in maintaining antibacterial efficacy regardless of the biofilm development stage, highlighting its potential utility in oral care.

Physiological and genetic responses of the benthic dinoflagellate Prorocentrum lima to polystyrene microplastics

Microplastics are well known as contaminants in marine environments. With the development of biofilms, most microplastics will eventually sink and deposit in benthic environment. However, little research has been done on benthic toxic dinoflagellates, and the effects of microplastics on benthic dinoflagellates are unknown. Prorocentrum lima is a cosmopolitan toxic benthic dinoflagellate, which can produce a range of polyether metabolites, such as diarrhetic shellfish poisoning (DSP) toxins. In order to explore the impact of microplastics on marine benthic dinoflagellates, in this paper, we studied the effects of polystyrene (PS) on the growth and toxin production of P. lima. The molecular response of P. lima to microplastic stress was analyzed by transcriptomics. We selected 100 nm, 10 μm and 100 μm PS, and set three concentrations of 1 mg L−1, 10 mg L−1 and 100 mg L−1. The results showed that PS exposure had limited effects on cell growth, but increased the OA and extracellular polysaccharide content at high concentrations. After exposure to PS MPs, genes associated with DSP toxins synthesis, carbohydrate synthesis and energy metabolism, such as glycolysis, TCA cycle and pyruvate metabolism, were significantly up-regulated. We speculated that after exposure to microplastics, P. lima may increase the synthesis of DSP toxins and extracellular polysaccharides, improve the level of energy metabolism and gene expression of ABC transporter, thereby protecting algal cells from damage. Our findings provide new insights into the effects of microplastics on toxic benthic dinoflagellates.

Impacts of microplastic and seawater acidification on unicellular red algae: Growth response, photosynthesis, antioxidant enzymes, and extracellular polymer substances

Microplastics (MPs) pollution and seawater acidification have increasingly become huge threats to the ocean ecosystem. Their impacts on microalgae are of great importance, since microalgae are the main primary producers and play a critical role in marine ecosystems. However, the impact of microplastics and acidification on unicellular red algae, which have a unique phycobiliprotein antenna system, remains unclear. Therefore, the impacts of polystyrene-MPs alone and the combined effects of MPs and seawater acidification on the typical unicellular marine red algae Porphyridium purpureum were investigated in the current study. The result showed that, under normal seawater condition, microalgae densities were increased by 17.75-41.67 % compared to the control when microalgae were exposed to small-sized MPs (0.1 μm) at concentrations of 5-100 mg L-1. In addition, the photosystem II and antioxidant enzyme system were not subjected to negative effects. The large-sized MPs (1 μm) boosted microalgae growth at a low concentration of MPs (5 mg L-1). However, it was observed that microalgae growth was significantly inhibited when MPs concentration increased up to 50 and 100 mg L-1, accompanied by the remarkably reduced Fv/Fm value and the elevated levels of SOD, CAT enzymes, phycoerythrin (PE), and extracellular polysaccharide (EPS). Compared to the normal seawater condition, microalgae densities were enhanced by 52.11-332.56 % under seawater acidification, depending on MPs sizes and concentrations, due to the formed CO2-enrichment condition and appropriate pH range. PE content in microalgal cells was significantly enhanced, but SOD and CAT activities as well as EPS content markedly decreased under acidification conditions. Overall, the impacts of seawater acidification were more pronounced than MPs impacts on microalgae growth and physiological responses. These findings will contribute to a substantial understanding of the effects of MPs on marine unicellular red microalgae, especially in future seawater acidification scenarios.

Antibacterial Activity and Mechanism of Three Root Exudates from Mulberry Seedlings against Ralstonia pseudosolanacearum.

Bacterial wilt is a significant soil-borne disease that poses a threat to mulberry production yield and quality of agricultural production worldwide. However, the disease resistance mechanisms dependent on root exudates are not well understood. In this present study, we investigated the antibacterial mechanisms of the main active substances (erucamide, oleamide, and camphor bromide) present in mulberry root exudates (MRE) against Ralstonia pseudosolanacearum (Rp), the causal agent of bacterial wilt. Our findings revealed that these three active substances inhibited the growth activity of Rp by affecting the cell morphology and extracellular polysaccharide content, as well as triggering a burst of reactive oxygen species. The active substances induced oxidative stress, leading to a decrease in Rp growth. Additionally, the expression levels of key genes in the hrp gene cluster (hrpB, hrpX, and hrpF) and other virulence-related genes (such as ripAW, ripAE, Rs5-4819, Rs5-4374, ace, egl3, and pehB) were significantly reduced upon treatment with the active substances. Further pathogenicity experiments demonstrated that root exudates (at a concentration of 1.5 mg·mL-1) delayed or slowed down the occurrence of bacterial wilt in mulberry. These findings provide valuable insight into the antimicrobial mechanisms of MRE against Rp and lay a theoretical foundation for the development and application of biocontrol agents to control mulberry bacterial wilt.

BolA promotes the generation of multicellular behavior in S. typhimurium by regulating the c-di-GMP pathway genes yeaJ and yhjH

The generation of multicellular behavior enhances the stress adaptability, antibiotic resistance, and pathogenic potential of Salmonella enterica serovar Typhimurium (S. typhimurium), which is challenging for its prevention and control. Therefore, determination of the mechanism of multicellular behavior development is urgently required. Accordingly, this study investigated BolA, a transcription factor that promotes bacterial survival under different stresses. We found that BolA promoted the generation of multicellular behavior. Furthermore, transcriptome analysis revealed that BolA affected the expression of numerous genes, including biofilm formation and motility-related genes. In terms of biofilm formation, compared with the wild-type strain, bolA overexpression (269BolA+) increased the extracellular matrix content (extracellular polysaccharide, extracellular protein, and extracellular DNA (eDNA) by upregulating gene expression, ultimately increasing the biofilm formation ability by 2.56 times. For motility, bolA overexpression inhibited the expression of flagella synthesis genes, resulting in a 91.15 % decrease in motility compared with the wild-type (6 h). Further mechanistic analysis demonstrated that BolA affected the expression of the C-di-GMP pathway genes yeaJ and yhjH, which influenced the generation of multicellular behavior. In terms of biofilms, the extracellular polysaccharide content of 269BolA + ∆Yeaj (bolA overexpression and yeaJ deletion) was reduced by 89.91 % compared with 269BolA+, resulting in a 71.1 % reduction in biofilm forming ability. The motility of the 269∆BolA∆Yhjh (bolA/yhjH double deletion) strain was significantly decreased compared with that of 269∆BolA. Finally, the LacZ gene reporting showed that BolA promoted and inhibited the expression of yeaJ and yhjH, respectively. In conclusion, BolA mainly improves the content of extracellular polysaccharide by promoting the expression of yeaJ, thus enhancing the formation of biofilms. BolA also restricts flagellar synthesis by inhibiting yhjH expression, therefore reducing motility, ultimately promoting multicellular behavior arises. These findings lay a theoretical foundation for the prevention and control of S. typhimurium.

Improvement of extracellular polysaccharides production from Cordyceps militaris immobilized alginate beads in repeated-batch fermentation

Cordyceps militaris (C. militaris) is a unique medicinal fungus of the genus Cordyceps. It has high economic value due to various biological functions from the extracellular polysaccharides (EPSs) it produces. The aim of this study was to establish a mycelium-immobilization system for EPS production in a bubble column system. First, EPS production was optimized by 90.2% (from 0.72 to 1.37 g/L) using response surface methodology under an alginate-immobilized mycelium system with 6 days of cultivation. The highest EPS production was obtained (5.3 g/L) in a flask system through elongation to 15 days of cultivation. EPS production subsequently further increased to 10.42 g/L utilizing a bubble column reactor. Furthermore, the system can be reused for at least three rounds. In the EPS characteristics analysis, the produced EPSs were mainly composed of glucose and galactose. They presented strong antibacterial activity against Staphylococcus aureus. In terms of antioxidant activity, respective the EPS concentration that caused 50% scavenging of DPPH and ABTS free radicals values (IC50) were 4.65 and 7.92 mg/mL. In summary, these findings should be helpful for establishing a suitable industrial fermentation system for C. militaris and can be studied in the food and medical industries in future work.

The extracellular polysaccharide determine the physico-chemical surface properties of Microcystis.

Microcystis possesses the capacity to form colonies and blooms in lakes and reservoirs worldwide, causing significant ecological challenges in aquatic ecosystems. However, little is known about the determining factors of physico-chemical surface properties that govern the competitive advantage of Microcystis. Here, The physico-chemical surface properties of Microcystis wesenbergii and Microcystis aeruginosa, including specific surface area (SSA), hydrophobicity, zeta potential, and functional groups were investigated. Additionally, the extracellular polysaccharide (EPS) were analyzed. Laboratory-cultured Microcystis exhibited hydrophilic, a negative zeta potential and negatively charged. Furthermore, no significant relationship was shown between these properties and the cultivation stage. Microcystis wesenbergii exhibited low free energy of cohesion, high surface free energy, high growth rate, and high EPS content during the logarithmic phase. On the other hand, M. aeruginosa displayed lower free energy of cohesion, high surface free energy, high EPS content, and high growth rate during the stationary phase. These characteristics contribute to their respective competitive advantage. Furthermore, the relationship between EPS and surface properties was investigated. The polysaccharide component of EPS primarily influenced the SSA and total surface energy of Microcystis. Likewise, the protein component of EPS influenced hydrophobicity and surface tension. The polysaccharide composition, including glucuronic acid, xylose, and fructose, mainly influenced surface properties. Additionally, hydrophilic groups such as O-H and P-O-P played a crucial role in determining hydrophobicity in Microcystis. This study elucidates that EPS influenced the SSA, hydrophobicity, and surface free energy of Microcystis cells, which in turn impact the formation of Microcystis blooms and the collection.

Bioinspired total synthesis and biological activity of Pegaharine A.

Phytopathogens cause various diseases by parasitizing crops, reducing crop yield and resulting in substantial economic losses in agricultural production. A novel type isolated from the perennial herbaceous Peganum harmala L. seeds, β-carboline alkaloids pegaharine A (PA), has become a hot topic in developing plant-originated green pesticides owing to their significant physiological activities. A scalable bioinspired total synthesis of PA is accomplished in the present work. The systematical biological assay study showed that PA exhibited moderate inhibitory activity against nine tested plant pathogenic fungi and showed significant inhibitory activity in vitro against the three tested plant pathogenic bacteria. Most noteworthy is the inhibitory rates of PA on Xanthomonas oryzae pv. oryzae (Xoo), X. oryzae pv. oryzicola (Xoc) and X. axonopodis pv. citri (Xac) of 93.6%, 92.1% and 86.1%, respectively, which are better than the control drug, bismerthiazol (63.4%, 61.2% and 53.7% at 100 μg mL-1 concentration). Furthermore, the EC50 value of PA against Xoo, Xoc and Xac was 52.2, 60.0 and 65.1 μg mL-1 , respectively, superior to 72.9, 64.2 and 70.1 μg mL-1 of the control drug. Moreover, the anti-Xoo mechanistic studies revealed that PA exerted its antibacterial effects by increasing the permeability of the bacterial membrane, reducing the extracellular polysaccharide content and inducing morphological changes in bacterial cells. A novel β-carboline alkaloid, PA, was prepared by biomimetic total synthesis. Its significant antibacterial activity was closely related to the permeation of bacterial cell membranes, which was confirmed by anti-Xoo mechanistic studies. More importantly, the structure could be regarded as a model for developing novel bactericides. © 2023 Society of Chemical Industry.

A novel antibacterial and anti-biofilm material: Biosynthetic silver nanoparticles mediated by extract from Oplopanax elatus adventitious roots

The search for plant resources involved in the biosynthesis of silver nanoparticles (AgNPs) is crucial for the diversity of AgNP materials. Oplopanax elatus is a valuable medicinal shrub and its adventitious root (AR) culture is a modern method for producing the raw material. To explore an efficient biosynthetic AgNPs, this study synthesized AgNPs using O. elatus AR extract (OE-AgNPs) and the characterization of OE-AgNPs was performed. Finally, the antibacterial and anti-biofilm effects of OE-AgNPs were investigated. The result showed that OE-AgNPs had an obvious absorption peak of nanosilver with a spherical shape and a silver-like face-centered cubic structure, which the OE-AgNPs were capped by compounds with aliphatic amine, amide, carbonyl, and hydroxyl groups of O. elatus AR extract, indicating a successful synthesis of OE-AgNPs. The 10–40 nm particles were the most abundant in the OE-AgNPs, accounting for 70%. OE-AgNPs had the strongest antibacterial activity against Escherichia coli and Pseudomonas aeruginosa among six bacterial species tested, with a minimum inhibitory concentration (MIC) of 0.75 μg/mL, and the cell contents of E. coli and P. aeruginosa leaked out by OE-AgNP treatment, and bacterial respiration was inhibited. After OE-AgNP treatment, biofilms of E. coli and P. aeruginosa were efficiently scavenged with a scavenging rate of more than 60%; the extracellular polysaccharide concentrations in biofilms were also ruduced. The findings of this study suggested that OE-AgNPs could be used as a potential antibacterial and antibiofilm substance to apply in the product production and indicated the feasibility of expanding the application of O. elatus ARs.

The role of extracellular polysaccharide produced by Bradyrhizobium strain in root growth, improvement of soil aggregate stability and reduction of soil detachment capacity

It is well demonstrated that the production of extracellular polysaccharide by bacteria, as biological binding agent, can increase soil aggregate stability. Also, plant roots have significant effects on the soil erosion rates, since they can strongly change the soil detachment capacity (Dc). This study quantifies Dc at different flow rates in soils treated with biofertilizer under four species (Alnus subcordata, Brachythecium plumose, Gleditsia caspica and Sambucus ebulus species) in the Saravan forest park of Iran and develops regression equations for predicting Dc. Undisturbed samples collected from soils with the four tree species and subjected to five slopes (from 4.2 to 38.3%) and five water discharges (from 0.28 to 0.71 L m−1 s−1) using a hydraulic flume. The results showed that Dc was significantly lower in soils with Alnus subcordata species compared to Brachythecium plumose, Gleditsia caspica and Sambucus ebulus species, as the consequence of the changes in the root characteristics, mean weight diameter (MWD) and macronutrients, so that Dc was negatively correlated with root weight density, root length and root biomass. A linear regression model was found for MWD as a function of extracellular polysaccharide concentration (R2 = 0.74). The unit stream power had high accuracy for predicting Dc for all of the studied species. The lowest value of rill erodibility (Kr) was obtained in soils with Alnus subcordata species using regression relationship between the Dc and the shear stress of the soil. This experiment helped to show the importance of microorganisms in reducing the soil detachment rates and provided a contribution in understanding the choice of appropriate species for soil conservation.

The potential use of glycosyl-transferase inhibitors for targeted reduction of S. mutans biofilms in dental materials

Streptococcus mutans is the primary oral caries-forming bacteria, adept at producing “sticky” biofilms via the synthesis of insoluble extracellular polysaccharides (EPS), catalyzed by glucosyltransferases (GTFs). To circumvent the use of broad-spectrum antibiotics to combat these bacteria, this study sought to modify existing EPS-targeting small molecules with the ultimate goal of producing anti-biofilm polymer surfaces specifically targeting S. mutans. To achieve this, a known GTF inhibitor (G43) was modified with methoxy or tetraethyleneglycol substitutions in different positions (nine derivatives, tested at 50-µM) to pinpoint potential sites for future methacrylate functionalization, and then assessed against single-species S. mutans biofilms. As expected, the compounds did not diminish the bacterial viability. In general, the compounds with methoxy substitution were not effective in reducing EPS formation, whereas the tetraethyleneglycol substitution (G43-C3-TEG) led to a decrease in the concentration of insoluble EPS, although the effect is less pronounced than for the parent G43. This aligns with the reduced GTF-C activity observed at different concentrations of G43-C3-TEG, as well as the consequent decrease in EPS formation, and notable structural changes. In summary, this study determined that G43-C3-TEG is non-bactericidal and can selectively reduce the biofilm formation, by decreasing the production of EPS. This molecule will serve to functionalize surfaces of materials to be tested in future research.

Enhancement of cyanobacterial blooms buoyancy by controlling extracellular polysaccharides content and cation concentration under light-limited condition

Enhancement of cyanobacterial blooms buoyancy by controlling extracellular polysaccharides content and cation concentration under light-limited condition

Effects of astragaloside IV on membrane biofouling in a membrane bioreactor from the perspective of extracellular polymeric substances and the bacteria community structure

At present, the control of membrane biofouling is mainly achieved by regulating quorum sensing. In this study, the quorum-sensing inhibitor astragaloside IV was used to prevent membrane biofouling in a membrane bioreactor. It was found that astragaloside IV at a concentration of 1 mg/L doubled the time taken for membrane contamination to occur, and decreased the energy required for filtration by 62% without affecting reactor’s processing performance. This is because the addition of astragaloside IV decreased the soluble microbial protein concentration and the loosely bound extracellular polysaccharide content. Biodiversity analysis indicated that astragaloside IV may have inhibited membrane contamination by interfering with the structure of the bacteria community, particularly the populations of quorum-sensing bacteria, bacteria related to extracellular polymeric substance production, and quorum-quenching bacteria. This is the first time the effects of astragaloside IV on membrane contamination in membrane reactors and the inhibition mechanisms involved were investigated. Using astragaloside IV could effectively decrease the energy consumed in operating a membrane bioreactor.

Antimicrobial behavior of titanium coating with chlorhexidine-doped thin film exposed to a biofilm supplemented with nicotine.

Because nicotine upregulates the growth of most oral bacteria, this in vitro study investigated the antimicrobial effect of chlorhexidine-doped thin film on commercially pure titanium against Fusobacterium nucleatum (F. nucleatum) biofilm supplemented with different concentrations of nicotine (0, 1, and 2 mg/mL). Biofilms were formed on a chlorhexidine-doped thin film on commercially-pure-titanium discs and compared to the control groups. Biofilm viability, total biofilm growth using a spectrophotometer, extracellular polysaccharide content, and pH variations were assessed as dependent variables. Data were submitted to ANOVA and Tukey honest significant difference tests (α=0.05). F. nucleatum biofilm growth was inhibited when exposed to chlorhexidine-doped thin film (p<0.05). Biofilm supplemented with nicotine did not impact the synthesis of EPS on the same type of treatment (p>0.05). The pH values were significantly increased with the increase of nicotine concentration (p<0.05). Chlorhexidine-doped thin film was effective in reducing F. nucleatum biofilm supplemented with nicotine.

Phosphorus level impacts luteolin effect on Microcystis aeruginosa growth and microcystin-pollution risk − Novel perspective from correlation between exopolymers substances fractions and microcystin-production/release

Luteolin as a phytogenic algicide can inhibit the growth and microcystins (MCs) release of Microcystis, a dominant genus during cyanobacterial blooms, but how phosphorus (P) level impacts luteolin effect on its growth and MC-pollution risk is unclear. By employing Microcystis aeruginosa as test alga, this study addressed this concern and explored response mechanisms from novel insights of relationship between extracellular polysaccharide (ex-poly) and protein (ex-pro) contents and MC-production/release. At each P level (0.05–5 mg/L), rising luteolin dose more greatly inhibited Microcystis growth and MC-pollution risk, with growth inhibition ratio of around 10%–30%, 20%–50% and 40%–90% for 3, 6 and 12 mg/L luteolin, respectively, but almost increasingly enhanced cellular ability of MC-production/conservation and total and bound ex-poly/ex-pro production. Rising P level promoted Microcystis growth and intracellular/extracellular MCs content (IMC, EMC) in test system at each luteolin dose, thus higher P level weakened algicidal and MC-removal effects of luteolin, indicating that P-decrease was required for stronger application outcome of luteolin. Total and bound ex-poly/ex-pro amount were positively correlated with cellular MC-production/conservation ability, IMC and EMC, which constituted cooperative stress-defense of Microcystis at each P level. Besides, rising luteolin dose posed stronger algicidal effect by inactivating gene expression involving peroxidase synthesis (especially at P-limitation), photosynthesis and P acquisition, while rising P level alleviated algicidal and MC-pollution inhibition effects of luteolin by enhancing gene expression involving N acquisition and peroxidase synthesis. This study shed novel insights for P-dependent effect and mechanisms of luteolin on toxigenic Microcystis growth and MC-pollution control, which guided to mitigating toxigenic Microcystis-dominated cyanobacterial blooms in different P-level water areas.

Transcriptomic analysis reveals the antibiofilm mechanism of Lacticaseibacillus rhamnosus MS1 against Vibrio parahaemolyticus

This study aimed to explore novel anti-biofilm agents from Lacticaseibacillus rhamnosus. We found that the ethyl acetate extract of L. rhamnosus MS1 effectively reduced the motility, extracellular polysaccharide (EPS) content and biofilm production of Vibrio parahaemolyticus and confirmed the anti-biofilm effect by light microscopy and scanning electron microscopy. Transcriptomic analysis revealed that there were 284 significantly up-regulated genes and 443 down-regulated genes in V. parahaemolyticus biofilm treated with the subinhibitory concentration of the extract of L. rhamnosus MS1. Quantitative real-time polymerase chain reaction (qRT-PCR) was conducted to verify the expression changes of the biofilm-related genes (cpxA, cysD, D0853_RS09350, D0853_RS15790, emrD, flgM, luxS, ompW, pyk and tdh), and the results indicated that L. rhamnosus MS1 regulated V. parahaemolyticus quorum sensing (QS), biofilm formation, virulence factors and related metabolic pathways. This study revealed the efficacy of L. rhamnosus MS1 as an anti-biofilm agent.

Achieving efficient biodegradation of hydrophobic toluene via the enhancement of β-cyclodextrin and atomization

Industrial production processes produce large amounts of toluene, endangering human health and air quality. The hydrophobicity of toluene limits the removal performance in biotrickling filters (BTFs). To improve the removal efficiency of toluene in BTFs, β-cyclodextrin (β-CD) coupled with atomization was employed in this study. The results showed that β-CD and atomization significantly enhanced the biodegradation of toluene and the maximum elimination capacity reached 90.9 g/(m3·h). At the optimized conditions that β-CD additional of 12.5 g/L and an atomizer with power of 19.8 W, the removal efficiency of toluene was up to 92.6 % (inlet concentration, 0.7 g/m3), which was 17 % higher than that of the control. The use of β-CD combined with atomization increased the extracellular protein and polysaccharide contents. The biofilm surface hydrophobicity of the experimental group reached 79 % which was 1.34 times that of the control. The abundance of toluene degrading bacteria, Pseudomonas sp. and Mycobacterium sp., significantly increased under β-CD with atomization. The addition of β-CD with atomization increased mass transfer, the relative hydrophobicity of biofilm, and the functional microbes so as to enhance treatment of toluene. This study may provide guidelines for the design and technological development of hydrophobic toluene control in industrial waste gas.

Shift of calcium-induced Microcystis aeruginosa colony formation mechanism: From cell adhesion to cell division

Colony formation is an essential stage of cyanobacterial blooms. High calcium concentration can promote Microcystis aeruginosa aggregation behavior, but the mechanism of colony formation caused by calcium has rarely been reported. In this study, high calcium-induced colony formation was identified as a shift from cell adhesion to cell division, rather than only cell adhesion as previously thought. Algae responded to this calcium-induced environmental pressure by aggregating and forming colonies. Algal cells initially secreted large quantities of extracellular polysaccharides (EPS) and rapidly aggregated by cell adhesion. The highest aggregation proportion was up to 68.93%. However, high calcium concentrations cannot completely inhibit algal cell growth, but only delay the algae into the rapid growth phase. With adaption to calcium and existing high EPS content, the daughter cells reduced EPS synthesis and the aggregation proportion decreased. The increasing growth rate was also responsible for the decreased xylose content in EPS. The mechanism of colony formation changed to cell division. The downregulation of genes related to EPS secretion also supported this hypothesis. Overall, these results can benefit for our understanding of cyanobacterial bloom formation.

Antibiofilm and Antiquorum Sensing Potential of Lactiplantibacillus plantarum Z057 against Vibrio parahaemolyticus.

Vibrio parahaemolyticus is a widespread foodborne pathogen that causes serious seafood-borne gastrointestinal infections. Biofilm and quorum sensing (QS) are critical in regulating these infections. In this study, first, the ability of Lactiplantibacillus plantarum Z057 to compete, exclude, and displace V. parahaemolyticus biofilm was evaluated. Then, the inhibitory effects of L. plantarum Z057 extract (Z057-E) on V. parahaemolyticus biofilm and QS were explored from the aspects of biofilm biomass, metabolic activity, physicochemical properties, extracellular polymer matrix content, QS signal AI-2 activity, biofilm microstructure, and the expression levels of biofilm and QS-related genes. Results showed that L. plantarum Z057 effectively inhibited biofilm formation of V. parahaemolyticus and interfered with the adhesion of V. parahaemolyticus on the carrier surface. In addition, the Z057-E could significantly reduce the biofilm biomass, metabolic activity, hydrophobicity, auto-aggregation ability, swimming and swarming migration diameter, AI-2 activity, extracellular polysaccharide (EPS), and extracellular protein content of V. parahaemolyticus. Fluorescence microscope and scanning electron microscope (SEM) images demonstrated that the Z057-E could efficiently inactivate the living cells, destroy the dense and complete biofilm architectures, and reduce the essential component of the extracellular polymer matrix. Real-time fluorescence quantitative PCR revealed that the Z057-E treatment down-regulated the expression of flagellum synthesis-related genes (flaA, flgM), EPS, and extracellular protein synthesis-related genes (cpsA, cpsQ, cpsR, ompW), QS-related genes (luxS, aphA, opaR), and hemolysin secretion-related genes (toxS, toxR) of V. parahaemolyticus. Thus, our results suggested that L. plantarum Z057 could represent an alternative biocontrol strategy against foodborne pathogens with anti-adhesive, antibiofilm, and antiquorum sensing activities.

Factors Influencing the Production of Extracellular Polysaccharides by the Green Algae Dictyosphaerium chlorelloides and Their Isolation, Purification, and Composition.

The freshwater green microalgae, Dictyosphaerium chlorelloides (CCALA 330), has the ability to produce extracellular polysaccharides (EPS). Conditions for optimum growth and EPS overproduction were determined in laboratory-scale tubular photobioreactors (PBR) with a working volume of 300 mL. Multiple limitations in nutrient supply were proven to be an effective method for EPS overproduction. Salinity stress was also applied to the culture, but no significant increase in EPS production was observed. The effects of different nitrogen sources were examined and the microalgae exhibited the fastest growth and EPS production in medium containing ammonium nitrate. Under determined optimal conditions, EPS concentration reached 10 g/L (71% of the total biomass) and a total biomass of 14 g/L at the end of 17 days cultivation. Pilot-scale cultivation was also carried out in a column type airlift photobioreactor (PBR) with a working volume of 60 L. A new and efficient methodology was developed for separating cells from the EPS-containing culture broth. Due to the strong attachment between cells and EPS, high-pressure homogenization was carried out before a centrifugation process. The EPS in the supernatant was subsequently purified using ultrafiltration. The green microalgae Dictyosphaerium chlorelloides may therefore be appropriate for the commercial production of EPS.