Biofilm Components Research Articles

The long and the short of Periscope Proteins

Bacteria sense, interact with, and modify their environmental niche by deploying a molecular ensemble at the cell surface. The changeability of this exposed interface, combined with extreme changes in the functional repertoire associated with lifestyle switches from planktonic to adherent and biofilm states necessitate dynamic variability. Dynamic surface changes include chemical modifications to the cell wall; export of diverse extracellular biofilm components; and modulation of expression of cell surface proteins for adhesion, co-aggregation and virulence. Local enrichment for highly repetitive proteins with high tandem repeat identity has been an enigmatic phenomenon observed in diverse bacterial species. Preliminary observations over decades of research suggested these repeat regions were hypervariable, as highly related strains appeared to express homologues with diverse molecular mass. Long-read sequencing data have been interrogated to reveal variation in repeat number; in combination with structural, biophysical and molecular dynamics approaches, the Periscope Protein class has been defined for cell surface attached proteins that dynamically expand and contract tandem repeat tracts at the population level. Here, I review the diverse high-stability protein folds and coherent interdomain linkages culminating in the formation of highly anisotropic linear repeat arrays, so-called rod-like protein 'stalks', supporting roles in bacterial adhesion, biofilm formation, cell surface spatial competition, and immune system modulation. An understanding of the functional impacts of dynamic changes in repeat arrays and broader characterisation of the unusual protein folds underpinning this variability will help with the design of immunisation strategies, and contribute to synthetic biology approaches including protein engineering and microbial consortia construction.

Group II truncated haemoglobin YjbI prevents reactive oxygen species-induced protein aggregation in Bacillus subtilis.

Oxidative stress-mediated formation of protein hydroperoxides can induce irreversible fragmentation of the peptide backbone and accumulation of cross-linked protein aggregates, leading to cellular toxicity, dysfunction, and death. However, how bacteria protect themselves from damages caused by protein hydroperoxidation is unknown. Here, we show that YjbI, a group II truncated haemoglobin from Bacillus subtilis, prevents oxidative aggregation of cell-surface proteins by its protein hydroperoxide peroxidase-like activity, which removes hydroperoxide groups from oxidised proteins. Disruption of the yjbI gene in B. subtilis lowered biofilm water repellence, which associated with the cross-linked aggregation of the biofilm matrix protein TasA. YjbI was localised to the cell surface or the biofilm matrix, and the sensitivity of planktonically grown cells to generators of reactive oxygen species was significantly increased upon yjbI disruption, suggesting that YjbI pleiotropically protects labile cell-surface proteins from oxidative damage. YjbI removed hydroperoxide residues from the model oxidised protein substrate bovine serum albumin and biofilm component TasA, preventing oxidative aggregation in vitro. Furthermore, the replacement of Tyr63 near the haem of YjbI with phenylalanine resulted in the loss of its protein peroxidase-like activity, and the mutant gene failed to rescue biofilm water repellency and resistance to oxidative stress induced by hypochlorous acid in the yjbI-deficient strain. These findings provide new insights into the role of truncated haemoglobin and the importance of hydroperoxide removal from proteins in the survival of aerobic bacteria.

Strategies for dispersion of cariogenic biofilms: applications and mechanisms.

Bacteria residing within biofilms are more resistant to drugs than planktonic bacteria. They can thus play a significant role in the onset of chronic infections. Dispersion of biofilms is a promising avenue for the treatment of biofilm-associated diseases, such as dental caries. In this review, we summarize strategies for dispersion of cariogenic biofilms, including biofilm environment, signaling pathways, biological therapies, and nanovehicle-based adjuvant strategies. The mechanisms behind these strategies have been discussed from the components of oral biofilm. In the future, these strategies may provide great opportunities for the clinical treatment of dental diseases. Graphical

Formation and influencing factors of disinfection by-products from bacterial materials in drinking water distribution systems

Abstract In recent years, the contribution of bacterial materials in water distribution systems to DBPs has attracted widespread attention from researchers. The risks and influencing factors of bacterial DBPs are reviewed. Factors affecting the generation of bacterial DBPs include the characteristics of bacterial materials, disinfection process, piping materials, and water quality. Among the major components of biofilms, proteins or amino acids have the greatest risk of DBPs. Among the commonly used disinfection methods, chlorine is more suitable for continuous disinfection of pipe networks than chloramine, but chlorine dioxide may be a better substitute in the future. Pipes with good biological stability, such as polyethylene (PE) pipe, should be encouraged to be used in water supply networks. Periodic removal of biofilms, reduction of organic matter and bromide ion concentrations, and maintenance of high flow rates are effective means to reduce bacterial DBPs. The control, removal and monitoring devices of tube wall biofilm need to be further developed and promoted. In the future, it is urgent to develop new disinfectants that are not easy to react with biomolecules and natural organic matter, and produce fewer or no by-products.

Rapid-killing efficacy substantiates the antiseptic property of the synergistic combination of carvacrol and nerol against nosocomial pathogens.

Globally, new classes of synthetic and natural antibiotics and antivirulents have continuously been validated for their potential broad-spectrum antagonistic activity with the aim of identifying an effective active molecule to prevent the spread of infectious agents in both food industry and medical field. In view of this, present study is aimed at evaluating the rapid killing efficacy of bioactive molecules Carvacrol (C) and Nerol (N) through British Standard European Norm 1276: phase2/step1 (EN1276) protocol. Active molecules C and N showed broad-spectrum antimicrobial activity against the test strains Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli and Enterococcus hirae at concentration range of 78.125, 625, 156.25 and 312.5 μg/mL, respectively, for C, and 625 μg/mL for N. Whereas, combinatorial approach showed efficient activity with four times reduced concentration of C and N at 78.125 and 156.25 µg/mL, respectively, against test strains. Further, EN1276 results proved the rapid killing efficacy of test strains in 1 min of contact time with significant (> 5 log) growth reduction at 100X concentration of actives. SEM analysis and reduced concentration of protease, lipids and carbohydrate contents of treated group biofilm components ascertained preformed biofilm disruption potential of C + N on polystyrene and nail surfaces. C + N at synergistic concentration exhibited no adverse effect on HaCaT cells at 78.125 µg/mL (C) + 156.25 µg/mL (N). Taken together, based on the observed experimental results, present study evidence the antiseptic/disinfectant ability of C + N and suggest that the combination can preferentially be used in foam-based hand wash formulations.Supplementary InformationThe online version contains supplementary material available at 10.1007/s00203-022-03197-x.

Buffering Capacity and Effects of Sodium Hexametaphosphate Nanoparticles and Fluoride on the Inorganic Components of Cariogenic-Related Biofilms In Vitro.

Despite the remarkable effects of sodium hexametaphosphate nanoparticles (HMPnano) on dental enamel de-/re-mineralization processes, information on the effects of these nanoparticles on biofilms is scarce. This study assessed the effects of HMPnano, with or without fluoride (F), on the inorganic components and pH of Streptococcus mutans and Candida albicans dual-species biofilms. Solutions containing conventional/micro-sized HMP (HMPmicro) or HMPnano were prepared at 0.5% and 1%, with or without 1100 ppm F. A 1100 ppm F solution and pure artificial saliva were tested as positive and negative controls, respectively. The biofilms were treated three times and had their pH analyzed, and the concentrations of F, calcium, phosphorus, and HMP in the biofilm biomass and fluid were determined. In another set of experiments, after the last treatment, the biofilms were exposed to a 20% sucrose solution, and the biofilm pH and inorganic components were evaluated. The 1% HMPnano solution with F led to the highest biofilm pH, even after exposure to sucrose. The 1% HMPnano solution without F led to significantly higher phosphorus concentrations in comparison to all other groups. It can be concluded that 1% HMPnano and F influenced the biofilm pH, besides affecting most of the inorganic components of the dual-species biofilms.

Novel Experimental In-Office Bleaching Gels Containing Co-Doped Titanium Dioxide Nanoparticles.

The present study reports on the development and testing of novel bleaching agents containing co-doped metaloxide nanoparticles (NP; 0%, 5%, 10% v/w) and hydrogen peroxide (HP, 0%, 6%, 15%, and 35%). Bovine blocks (n = 200, A = 36 mm2) were obtained and randomly distributed into experimental groups (n = 10/group). NPs were incorporated into gels before bleaching (3 sessions, 7 days apart, 30 min/session, irradiated with violet light-LT). Color changes (ΔE00, ΔWID), mineral content (CO32−, PO43−), and topography were assessed (spectrophotometer, ATR-FTIR, and AFM) before and after bleaching procedures (14 days). Metabolic status and three-dimensional components of non-disrupted Streptococcus mutans biofilms were investigated using a multimode reader and confocal microscopy. The results indicate that ΔE00 and ΔWID significantly increased with NPs’ concentrations and LT. The enamel’s mineral ratio was adversely impacted by HP, but alterations were less pronounced when using NP-containing gels. The enamel’s topography was not damaged by the bleaching protocols tested. The bioluminescence results show that bleaching protocols do not render latent antibacterial properties to enamel, and the confocal microscopy results demonstrate that the 3-dimensional distribution of the components was affected by the protocols. The proposed nanotechnology improved the bleaching efficacy of experimental materials independent of hydrogen peroxide or irradiation and did not adversely impact the enamel’s surface properties or its chemical content.

Fluorescence lectin binding analysis of carbohydrate components in dental biofilms grown in situ in the presence or absence of sucrose.

Carbohydrate components, such as glycoconjugates and polysaccharides, are constituents of the dental biofilm matrix that play an important role in biofilm stability and virulence. Exopolysaccharides in Streptococcus mutans biofilms have been characterized extensively, but comparably little is known about the matrix carbohydrates in complex, in situ-grown dental biofilms. The present study employed fluorescence lectin binding analysis (FLBA) to investigate the abundance and spatial distribution of glycoconjugates/polysaccharides in biofilms (n=306) from 10 participants, grown in situ with (SUC) and without (H2O) exposure to sucrose. Biofilms were stained with 10 fluorescently labeled lectins with different carbohydrate specificities (AAL, ABA, ASA, HPA, LEA, MNA-G, MPA, PSA, VGA and WGA) and analyzed by confocal microscopy and digital image analysis. Microbial composition was determined by 16S rRNA gene sequencing. With the exception of ABA, all lectins targeted considerable matrix biovolumes, ranging from 19.3% to 194.0% of the microbial biovolume in the biofilms, which illustrates a remarkable variety of carbohydrate compounds in in situ-grown dental biofilms. MNA-G, AAL, and ASA, specific for galactose, fucose, and mannose, respectively, stained the largest biovolumes. AAL and ASA biovolumes were increased in SUC biofilms, but the difference was not significant due to considerable biological variation. SUC biofilms were enriched in streptococci and showed reduced abundances of Neisseria and Haemophilus spp., but no significant correlations between lectin-stained biovolumes and bacterial abundance were observed. In conclusion, FLBA demonstrates the presence of a voluminous biofilm matrix comprising a variety of different carbohydrate components in complex, in situ-grown dental biofilms.

Bioaccessibility of Microplastic-Associated Antibiotics in Freshwater Organisms: Highlighting the Impacts of Biofilm Colonization via an In Vitro Protocol.

Microplastics in the environment can be colonized by microbes capable of forming biofilms, which may act as reactive coatings to affect the bioaccessibility of pollutants in organisms. This study investigated the dynamic evolution of biofilm colonization on microplastics and its impacts and mechanisms on the bioaccessibility of microplastic-associated sulfamethazine (SMT) via microcosm incubation in surface water and sediment. After 60 days of incubation, the microbial communities formed in microplastics were distinct and more diverse than those untethered in surroundings, and photoaging treatment decreased the affinity of biofilms on microplastics due to decreased hydrophobicity. Biofilm formation further enhanced the desorption and bioaccessibility of microplastic-sorbed SMT in organisms. In vitro experiments indicated that the critical effects were mainly related to the stronger interaction of gastrointestinal components (i.e., pepsin, bovine serum albumin (BSA), and NaT) with biofilm components (e.g., extracellular polymer substances) than with the pure surface of microplastics, which competed for binding sites in microplastics for SMT more significantly. Photoaging decreased the enhancing effects of biofilms due to their lower accumulation in aged microplastics. This study is the first attempt to reveal the role of biofilms in the bioaccessibility of microplastics with associated antibiotics and provide insights into the combined risk of microplastics in the environment.

Effects of Sodium Hexametaphosphate and Fluoride on the pH and Inorganic Components of Streptococcus mutans and Candida albicans Biofilm after Sucrose Exposure

In order to improve the anticaries effects of fluoridated products, the supplementation of these products has been considered a promising alternative for caries control. This study evaluated the effects of sodium hexametaphosphate (HMP) and/or fluoride (F) on the inorganic components and pH of Streptococcus mutans and Candida albicans dual-species biofilms. The biofilms were treated 72, 78, and 96 h after the beginning of their formation with 0.25, 0.5, or 1% HMP-containing solutions with or without F (500 ppm, as sodium fluoride). F-containing solutions (500 ppm and 1100 ppm) and artificial saliva were used as controls. The biofilms were exposed to a 20% sucrose solution after the third treatment. Along with the biofilm pH, the concentrations of F, calcium, phosphorus (P), and HMP were determined. HMP, combined with F, increased F levels and decreased P levels in the biofilm fluid compared to that of the solution with 500 ppm F. Exposure to sucrose decreased the concentrations of all ions in the biomass, except for HMP; 1% HMP, combined with F, promoted the highest pH. It can be concluded that HMP affected the inorganic composition of the biofilm and exerted a buffering effect on the biofilm pH.

Sorption of Cellulases in Biofilm Enhances Cellulose Degradation by Bacillus subtilis.

Biofilm commonly forms on the surfaces of cellulosic biomass but its roles in cellulose degradation remain largely unexplored. We used Bacillus subtilis to study possible mechanisms and the contributions of two major biofilm components, extracellular polysaccharides (EPS) and TasA protein, to submerged biofilm formation on cellulose and its degradation. We found that biofilm produced by B. subtilis is able to absorb exogenous cellulase added to the culture medium and also retain self-produced cellulase within the biofilm matrix. The bacteria that produced more biofilm degraded more cellulose compared to strains that produced less biofilm. Knockout strains that lacked both EPS and TasA formed a smaller amount of submerged biofilm on cellulose than the wild-type strain and also degraded less cellulose. Imaging of biofilm on cellulose suggests that bacteria, cellulose, and cellulases form cellulolytic biofilm complexes that facilitate synergistic cellulose degradation. This study brings additional insight into the important functions of biofilm in cellulose degradation and could potentiate the development of biofilm-based technology to enhance biomass degradation for biofuel production.

RapD Is a Multimeric Calcium-Binding Protein That Interacts With the Rhizobium leguminosarum Biofilm Exopolysaccharide, Influencing the Polymer Lengths

Rhizobium leguminosarum synthesizes an acidic polysaccharide mostly secreted to the extracellular medium, known as exopolysaccharide (EPS) and partially retained on the bacterial surface as a capsular polysaccharide (CPS). Rap proteins, extracellular protein substrates of the PrsDE type I secretion system (TISS), share at least one Ra/CHDL (cadherin-like) domain and are involved in biofilm matrix development either through cleaving the polysaccharide by Ply glycanases or by altering the bacterial adhesive properties. It was shown that the absence or excess of extracellular RapA2 (a monomeric CPS calcium-binding lectin) alters the biofilm matrix’s properties. Here, we show evidence of the role of a new Rap protein, RapD, which comprises an N-terminal Ra/CHDL domain and a C-terminal region of unknown function. RapD was completely released to the extracellular medium and co-secreted with the other Rap proteins in a PrsDE-dependent manner. Furthermore, high levels of RapD secretion were found in biofilms under conditions that favor EPS production. Interestingly, size exclusion chromatography of the EPS produced by the ΔrapA2ΔrapD double mutant showed a profile of EPS molecules of smaller sizes than those of the single mutants and the wild type strain, suggesting that both RapA2 and RapD proteins influence EPS processing on the cell surface. Biophysical studies showed that calcium triggers proper folding and multimerization of recombinant RapD. Besides, further conformational changes were observed in the presence of EPS. Enzyme-Linked ImmunoSorbent Assay (ELISA) and Binding Inhibition Assays (BIA) indicated that RapD specifically binds the EPS and that galactose residues would be involved in this interaction. Taken together, these observations indicate that RapD is a biofilm matrix-associated multimeric protein that influences the properties of the EPS, the main structural component of the rhizobial biofilm.

Neutralization of ionic interactions by dextran-based single-chain nanoparticles improves tobramycin diffusion into a mature biofilm

The extracellular matrix protects biofilm cells by reducing diffusion of antimicrobials. Tobramycin is an antibiotic used extensively to treat P. aeruginosa biofilms, but it is sequestered in the biofilm periphery by the extracellular negative charge matrix and loses its efficacy significantly. Dispersal of the biofilm extracellular matrix with enzymes such as DNase I is another promising therapy that enhances antibiotic diffusion into the biofilm. Here, we combine the charge neutralization of tobramycin provided by dextran-based single-chain polymer nanoparticles (SCPNs) together with DNase I to break the biofilm matrix. Our study demonstrates that the SCPNs improve the activity of tobramycin and DNase I by neutralizing the ionic interactions that keep this antibiotic in the biofilm periphery. Moreover, the detailed effects and interactions of nanoformulations with extracellular matrix components were revealed through time-lapse imaging of the P. aeruginosa biofilms by laser scanning confocal microscopy with specific labeling of the different biofilm components.

Biofilm formation of Salmonella enterica serovar Enteritidis cocultured with Acanthamoeba castellanii responds to nutrient availability.

Acanthamoeba spp. and Salmonella share common habitats, and their interaction may influence the biofilm-forming ability of Salmonella. In this study, biofilm formation of Salmonella enterica serovar Enteritidis cocultured with Acanthamoeba castellanii was examined in nutrient-rich and nutrient-deficient media. Furthermore, transcript copy number of biofilm-related genes in the biofilm cells of S. Enteritidis in monoculture was compared to those in coculture with A. castellanii. Results demonstrated that the presence of A. castellanii in the culture media activates the genes involved in the biofilm formation of S. Enteritidis, regardless of the nutrient availability. However, biofilm formation of S. Enteritidis cocultured with A. castellanii was not consistent with the transcript copy number results. In nutrient-rich medium, the number of Salmonella biofilm cells and the contents of the three main components of the biofilms including eDNA, protein, and carbohydrates were higher in the presence of A. castellanii compared to monocultures. However, in nutrient-deficient medium, the number of biofilm cells, and the amount of biofilm components in coculture conditions were less than the monocultures. These results indicate that despite activation of relevant genes in both nutrient-rich and nutrient-deficient media, biofilm formation of S. Enteritidis cocultured with A. castellanii responds to nutrient availability.

Probing the growth and mechanical properties of Bacillus subtilis biofilms through genetic mutation strategies

Bacterial communities form biofilms on various surfaces by synthesizing a cohesive and protective extracellular matrix, and these biofilms protect microorganisms against harsh environmental conditions. Bacillus subtilis is a widely used experimental species, and its biofilms are used as representative models of beneficial biofilms. Specifically, B. subtilis biofilms are known to be rich in extracellular polymeric substances (EPS) and other biopolymers such as DNA and proteins like the amyloid protein TasA and the hydrophobic protein BslA. These materials, which form an interconnected, cohesive, three-dimensional polymer network, provide the mechanical stability of biofilms and mediate their adherence to surfaces among other functional contributions. Here, we explored how genetically-encoded components specifically contribute to regulate the growth status, mechanical properties, and antibiotic resistance of B. subtilis biofilms, thereby establishing a solid empirical basis for understanding how various genetic engineering efforts are likely to affect the structure and function of biofilms. We noted discrete contributions to biofilm morphology, mechanical properties, and survival from major biofilm components such as EPS, TasA and BslA. For example, EPS plays an important role in maintaining the stability of the mechanical properties and the antibiotic resistance of biofilms, whereas BslA has a significant impact on the resolution that can be obtained for printing applications. This work provides a deeper understanding of the internal interactions of biofilm components through systematic genetic manipulations. It thus not only broadens the application prospects of beneficial biofilms, but also serves as the basis of future strategies for targeting and effectively removing harmful biofilms.

Regulation of Biofilm Exopolysaccharide Biosynthesis and Degradation in Pseudomonas aeruginosa.

Microbial communities enmeshed in a matrix of macromolecules, termed as biofilms, are the natural setting of bacteria. Exopolysaccharide is a critical matrix component of biofilms. Here, we focus on biofilm matrix exopolysaccharides in Pseudomonas aeruginosa. This opportunistic pathogen can adapt to a wide range of environments and can form biofilms or aggregates in a variety of surfaces or environments, such as the lungs of people with cystic fibrosis, catheters, wounds, and contact lenses. The ability to synthesize multiple exopolysaccharides is one of the advantages that facilitate bacterial survival in different environments. P. aeruginosa can produce several exopolysaccharides, including alginate, Psl, Pel, and lipopolysaccharide. In this review, we highlight the roles of each exopolysaccharide in P. aeruginosa biofilm development and how bacteria coordinate the biosynthesis of multiple exopolysaccharides and bacterial motility. In addition, we present advances in antibiofilm strategies targeting matrix exopolysaccharides, with a focus on glycoside hydrolases.

Biofilm-inspired Amyloid-Polysaccharide Composite Materials

Natural materials can inspire the design of functional structures and sustainable materials. Bacterial biofilms formed by polymicrobial cultures typically comprise a robust interwoven fibrous network made of proteins and exopolysaccharides, protecting the cells trapped inside against various environmental stressors. Inspired by these structures, we report an amyloid-polysaccharide composite matrix using the building block of Escherichia coli biofilms (CsgA) and chitin, the second most abundant polysaccharide. We chose these two substances to mimic the main components of biofilms and combine the toughness of polysaccharide fibers and the functionality of amyloid fusion proteins to produce a new functional hybrid material. Due to the chitin-binding domain and other functional tags, the chitin-amyloid composite matrix exhibited good molecular interaction and functionality, which formed a coating on the material surface and was made into a freestanding film. With the help of this composite coating, we adhered inorganic materials onto various material surfaces. The ability to interact with inorganic materials allows this new material to be applied to electronics and organic-inorganic interfaces, and the freestanding films provide an opportunity for fabricating intelligent materials or wearable electronic devices. Additionally, these biopolymer ingredients may offer a low-cost and environmentally-friendly bio-based alternative to industrial polymers or plastics in consumer products.

Impedance Characteristics of Monolayer and Bilayer Graphene Films with Biofilm Formation and Growth.

Biofilms are the result of bacterial activity. When the number of bacteria (attached to materials’ surfaces) reaches a certain threshold value, then the bacteria simultaneously excrete organic polymers (EPS: extracellular polymeric substances). These sticky polymers encase and protect the bacteria. They are called biofilms and contain about 80% water. Other components of biofilm include polymeric carbon compounds such as polysaccharides and bacteria. It is well-known that biofilms cause various medical and hygiene problems. Therefore, it is important to have a sensor that can detect biofilms to solve such problems. Graphene is a single-atom-thick sheet in which carbon atoms are connected in a hexagonal shape like a honeycomb. Carbon compounds generally bond easily to graphene. Therefore, it is highly possible that graphene could serve as a sensor to monitor biofilm formation and growth. In our previous study, monolayer graphene was prepared on a glass substrate by the chemical vapor deposition (CVD) method. Its biofilm forming ability was compared with that of graphite. As a result, the CVD graphene film had the higher sensitivity for biofilm formation. However, the monolayer graphene has a mechanical disadvantage when used as a biofilm sensor. Therefore, for this new research project, we prepared bilayer graphene with high mechanical strength by using the CVD process on copper substrates. For these specimens, we measured the capacitance component of the specimens’ impedance. In addition, we have included a discussion about the possibility of applying them as future sensors for monitoring biofilm formation and growth.

Punicalagin inhibits biofilm formation and virulence gene expression of Vibrioparahaemolyticus

Vibrio parahaemolyticus is a major food-borne pathogen and the leading cause of seafood-associated enteric infections worldwide. In addition, imprudent use of antibiotics has contributed to increasing antibiotic resistance in V. parahaemolyticus. Therefore, there is an urgent need to develop novel control strategies to reduce V. parahaemolyticus contamination and infection. V. parahaemolyticus could form biofilm on different surfaces and contains several virulence factors, which together contribute to its persistence and pathogenicity. In this study, anti-biofilm and potential anti-virulence effects of punicalagin against V. parahaemolyticus were investigated. Punicalagin at sub-inhibitory concentrations (3.125–12.5 μg/mL) exerted significant anti-biofilm activity (20.9–74.4% reduction). It decreased bacterial motility, and attenuated metabolic activity of cells inside biofilm. Meanwhile, punicalagin inhibited EPS production and caused reduction of exopolysaccharides in biofilms, and decreased Raman peak intensities of different biofilm components. Moreover, punicalagin significantly inhibited V. parahaemolyticus adhesion to Caco-2 cells (23.8–68.1% reduction) and downregulated expression of genes involved in V. parahaemolyticus biofilm formation and virulence. These findings suggest that punicalagin could be potentially developed as an alternative strategy to control V. parahaemolyticus biofilms in food industry and mitigate diseases caused by this pathogen.

Characterization of biofilms formed on polystyrene microplastics (PS-MPs) on the shore of the Tuul River, Mongolia

Microplastic (MP) surfaces are common sites for microbial colonization and promote biofilm formation in aquatic environments, resulting in changes to the surface properties of MPs and their interaction with pollutants. Although the diversity of microbial communities adhering to MPs has been well documented in aquatic environments, surface changes in MPs due to microbial colonization are still poorly understood. In this study, we aimed to evaluate the variations in the chemical structure and components of biofilms on the surface of polystyrene microplastics (PS-MPs) collected from the shore of the Tuul River in Mongolia, using micro-Fourier transform infrared (micro-FTIR) spectroscopy. We applied a spectral subtraction approach, and the differences in spectra between peroxide-treated and untreated PS-MP particles enabled us to obtain the structural features of biofilms that developed on the plastic surface. In addition, the surface photooxidation status of the sampled PS-MPs was calculated from the subtracted spectra of peroxide-treated and pristine PS-MPs. Various functional groups of N-containing organic substances from bacterial and fungal communities were detected in the obtained biofilm spectra. Based on the spectral characteristics, biofilm spectra were classified into four groups by applying principal component analysis (PCA). A wide range of carbonyl indices (CIs: 0.00–1.40) was found in the subtracted spectra between peroxide-treated and pristine PS-MPs, revealing that different levels of surface oxidation progressed by physical influences such as solar radiation and freeze-thaw cycles. Furthermore, lignocellulose and silicate were found on the PS-MP surface as allochthonous attachments. Considering the variation in residence time of PS-MPs, they attract plant residues and mineral particles through the development of biofilms and travel together in the river environment. Given that the dynamic behavior of MPs can be greatly affected by changes in their surfaces, further studies are needed to emphasize their link to organic matter dynamics.