Biofilm Matrix Research Articles

Chemical Composition and Phytotoxic and Antibiofilm Activity of the Essential Oils of Two Moroccan Retama Species.

This study reports chemical composition, phytotoxic and antibiofilm activities of essential oils (EOs) of R. dasycarpa and R. sphaerocarpa from Morocco. EOs were analyzed by GC/MS and their phytotoxicities were evaluated against germination and seedling growth of Lolium multiflorum, Sinapis alba and Raphanus sativus. The antimicrobial and anti-biofilm activities were studied against Gram-negative (Pseudomonas aeruginosa, Escherichia coli and Acinetobacter baumannii) and Gram-positive bacteria (Staphylococcus aureus and Listeria monocytogenes). Both EOs were abundant in oxygenated monoterpenes (40.01% and 23.57 %, respectively). Carvacrol is the predominant component in R. dasycarpa EO (17.80 %), and it also represents an appreciable amount in R. sphaerocarpa (8.96 %). R. sphaerocarpa showed total inhibition at high doses against all seeds. S. alba seeds were the most sensitive to all EOs. Minimum inhibitory concentration (MIC) values indicated significant inhibition for R. sphaerocarpa, between 24 and 30 μg/mL, with a remarkable antibacterial potential and biofilm formation inhibition. R. sphaerocarpa EO showed significant biofilm inhibition with variable efficacy depending on the strain and concentration, except for S. aureus. R. dasycarpa exhibited activity against all bacterial strains and effect on metabolism with activity also on mature biofilms. Results suggest that Retama EOs could have potential applications in the fields of food and health.

A new model of endotracheal tube biofilm identifies combinations of matrix-degrading enzymes and antimicrobials able to eradicate biofilms of pathogens that cause ventilator-associated pneumonia.

Ventilator-associated pneumonia is defined as pneumonia that develops in a patient who has been on mechanical ventilation for more than 48 hours through an endotracheal tube. It is caused by biofilm formation on the indwelling tube, which introduces pathogenic microbes such as Pseudomonas aeruginosa, Klebsiella pneumoniae and Candida albicans into the patient's lower airways. Currently, there is a lack of accurate in vitro models of ventilator-associated pneumonia development. This greatly limits our understanding of how the in-host environment alters pathogen physiology and the efficacy of ventilator-associated pneumonia prevention or treatment strategies. Here, we showcase a reproducible model that simulates the biofilm formation of these pathogens in a host-mimicking environment and demonstrate that the biofilm matrix produced differs from that observed in standard laboratory growth medium. In our model, pathogens are grown on endotracheal tube segments in the presence of a novel synthetic ventilated airway mucus medium that simulates the in-host environment. Matrix-degrading enzymes and cryo-scanning electron microscopy were employed to characterize the system in terms of biofilm matrix composition and structure, as compared to standard laboratory growth medium. As seen in patients, the biofilms of ventilator-associated pneumonia pathogens in our model either required very high concentrations of antimicrobials for eradication or could not be eradicated. However, combining matrix-degrading enzymes with antimicrobials greatly improved the biofilm eradication of all pathogens. Our in vitro endotracheal tube model informs on fundamental microbiology in the ventilator-associated pneumonia context and has broad applicability as a screening platform for antibiofilm measures including the use of matrix-degrading enzymes as antimicrobial adjuvants.

Comparative evaluation of antibacterial efficacy of nitrofurantoin, chitosan, and calcium hydroxide in combination with propylene glycol as an intracanal medicament against endodontic pathogen - An in vitro study.

The objective is to evaluate and compare the antibacterial efficacy of nitrofurantoin, chitosan, and calcium hydroxide (Ca(OH)2) in combination with propylene glycol (PG) as an intracanal medicament against endodontic pathogens. Fifty-two extracted single-rooted maxillary and mandibular anterior teeth and premolars were selected. The root canals were enlarged using Protaper universal rotary files. Clinical isolates of microorganisms collected from retreatment cases were used. Bacterial isolates obtained from infected root canals were introduced into brain-heart infusion (BHI) broth. Incubation of samples for 14 days was carried out to facilitate the development of mature biofilms. Intracanal medicaments were divided into four groups: Group 1 - Nirofurantoin+20% PG, Group 2 - Chitosan+20% PG, Group 3 - Ca(OH)2 + 20% PG, and Group 4 - 20% PG. The prepared root samples were incubated for 7 days. After collecting dentin samples, they were placed in a phosphate-buffered saline solution. Serial dilutions were then performed, and each dilution was plated on BHI agar. The plates were incubated for 24 h at 37°C. The antibacterial efficacy was assessed by calculating the percentage of remaining colony-forming units. Antibacterial efficacy of chitosan paste was significantly higher followed by nitrofurantoin as compared to other groups when used as an intracanal medicament. In the root-canal biofilm model, the combination of chitosan and PG demonstrated a significant reduction in the viability of endodontic pathogens when employed as intracanal medication for 7 days. This suggests its potential as an effective intracanal medicament for endodontic retreatment.

Enhancing Outcomes in Prosthetic Joint Infections: The Significance of the Periprosthetic Joint Infection Tumor, Node, and Metastasis (PJI-TNM) Classification and Biodegradable Antibiotic Beads.

Periprosthetic joint infections (PJIs) that occur after hip and knee arthroplasty have a major influence on patient outcomes and healthcare expenses. This study assesses the effectiveness of the PJI tumor, node, and metastasis (PJI-TNM) categorization system and the latest developments in local antibiotic delivery methods for the treatment of PJIs. The study involved a retrospective analysis of 23 patients who received treatment for septic hip or knee prostheses at the SUUB Orthopedics and Traumatology Clinic between January 1, 2022, and February 10, 2024. Approval was gained following ethical considerations. Patients were categorized using the PJI-TNM system, and their therapy was customized based on the severity of the infection. The surgical procedures involved either one-stage or two-stage revisions, utilizing vancomycin and gentamicin antibiotic-loaded calcium sulfate beads to administer antibiotics locally. Data pertaining to demographics, clinical characteristics, and microbiology were gathered and examined. The study comprised 14 male and 9 female patients, with an average age of 68 years. The presence of chronic infections was mostly seen, indicating the development of mature biofilm. Prevalent coexisting medical conditions included diabetes, obesity, and heart failure. The duration of infection control measures was, on average, six months, and 65% of patients reported experiencing enhanced mobility. Acute infections with positive antibiotic responses underwent one-stage modifications. For the majority of patients, a treatment approach involving two-stage modifications, which includes the use of antibiotic-loaded spacers followed by the installation of a prosthesis, proved to be beneficial. The PJI-TNM classification system improves the management of PJI by offering a systematic method for customized therapy. Calcium sulfate beads, which are biodegradable carriers for antibiotics, provide notable advantages, especially for individuals with severe comorbidities. Continuous progress in diagnostic techniques and localized administration of antibiotics is essential for enhancing the therapy of PJI and improving patient outcomes.

Antimicrobial impact of a propolis/PVA/chitosan composite and its prospective application against methicillin resistance bacterial infection

Seriously damaged skin could be infected by methicillin-resistant bacteria, which delays restoration. Propolis has bioactivity linked with its minor components, such as antimicrobials and antioxidants. Active sites in polyvinyl alcohol (PVA) and chitosan (CS) can enhance the nano-loading of natural extracts with activity amelioration. Korean propolis extract (KPE) loading to a nanocomposite possibly enhances its antimicrobial and anti-inflammatory potency. Composites were formed using two PVA/CS structures (1:1; 2:1), and their skin-application appropriateness was determined by mechanical properties, moisture content, water activity, and color. The composite of PVA/CS (1:1) was more practicable for KPE-loading. Increasing KPE concentrations (50, 100, 150, and 200 ng/mL) alters composite bioactivity measured by Fourier transmission infrared (FT-IR). Antibacterial potency of 200 ng KPE/mL was the most effective concentration, followed by 150 ng KPE/mL, against Staphylococcus aureus (MRSA) and Clostridium perfringens. The composite activity was measured as minimum inhibition (MIC) and minimum bacterial concentrations (MBC). At 200 ng KPE/mL, MIC and MBC against MRSA were 14.93 ± 1.21 and 20.21 ± 1.97 mg composite/mL, respectively. Significant inhibition was also recorded for antibiofilm formation, where MRSA growth was not detected after 4 hours of time intervals to the stainless-steel coupon. Compared to planktonic bacteria, the formed barrier of PVA/CS restrained the biofilm matrix formation and supported KPE antimicrobial. The impact of inhibition against biofilm formation depends on two parallel mechanisms (PVA barrier with hydrogen bonds, besides nano-KPE particle penetration into bacterial cells). The KPE-composite application to rats’ wounds shows significantly reduced MRSA infection. The results demonstrate the capability of KPE composite in reducing infection, healing correctly, and restoring hair. The wound swabbed test emphasizes this capacity, in which bacterial growth rate restriction was evaluated using a plate count assay. The results recommended 150 ng KPE/mL loading into CS/PVA (1:1) as an effective anti-pathogenic treatment, particularly against the MRSA infection of wounds.

Revision of ex vivo endodontic biofilm model using computer aided design

ObjectiveMost endodontic diseases are bacterium-mediated inflammatory or necrotic process induced by contaminated dental pulp. Although great advances are being performed to obtain more efficient antibacterial strategies for persistent infections, most studies lack of representative models to test their antibacterial effects and their outcomes cannot be promptly translated to clinical practice. Therefore, this study aimed to refine an ex vivo endodontic biofilm model combining human tooth, computer guided design and 3D printing to obtain a more reproducible and predictable model. MethodsMonoradicular teeth were cut using three different methods: hand-held (HCC), mechanical precision (MPC) and computer aid guided cutting (CGC). Then, blocks were reassembled. The different model preparations were assessed in terms of dimensional tolerance, surface analysis, liquid tightness and Enterococcus faecalis biofilm development for 21 days, which was studied by metabolic assays and confocal microscopy. Then, the proposed model was validated using different commercial disinfecting treatments. ResultsCGC exhibited significantly lower deviation and surface without defects compared to HHC and MPC, leading to superior liquid tightness. Similarly, mature biofilms with high metabolic activity and vitality were observed in all conditions, CGC showing the lowest variation. Regarding the model validation, all antibacterial treatments resulted in the complete eradication of bacteria in the standard 2D model, whereas commercial treatments exhibited varying levels of efficacy in the proposed ex vivo model, from moderately reduction of metabolic activity to complete elimination of biofilm. ConclusionsThe novel guided approach represents a more reliable, standardized, and reproducible model for the evaluation of endodontic disinfecting therapies. Clinical SignificanceDuring antibacterial treatment development, challenging 3D models using teeth substrates to test antibacterial treatments novel guided approach represents a more reliable, standardized, and reproducible model for the evaluation of endodontic disinfecting therapies.

Se-functionalized ZIF-8 nanoparticles: synthesis, characterization and disruption of biofilms and quorum sensing in Serratia marcescens

The majority of research on nanomaterials has been concentrated on metal nanoparticles since they are easily made and manipulated. Nanomaterials have shown a wide range of applications in biology. Nevertheless, their bioactivity declines due to their extreme susceptibility to and novel Se@ZIF-8 by chemical method. The sizes and morphologies of Se (0) and Se@ZIFchemical and physical stimuli. The goal of encapsulating these nanomaterials in a matrix is gradually being pursued, which boosts their affordability, stability, and usability. Metal-organic frameworks, often known as MOFs, have the potential to be the best platforms for encapsulating metal nanoparticles due to their well-defined frameworks, persistent porosity, and flexibility in modification. In this investigation, we report the synthesis and optimization of polyvinylpyrrolidone-stabilized Se(0) nanoparticles -8 were affected by the ratios of Se/Zn2+and [hmim]/Zn2+used. The optimized Se@ZIF-8 nanoparticles exhibited a particle size and zeta potential of 319 nm and -34 mv respectively. Transmission electron microscopy displayed spherical morphology for Se(0) nanoparticles, whereas the surface morphology of novel Se@ZIF-8 nanoparticles was drastically changed to hexagonal shaped structures with smooth surface morphologies in scanning electron microscopy (SEM). The DTA, TG/DTG, XRD analysis confirmed the presence of novel Se incorporated ZIF-8 nanoparticulate framework. The synthesized novel Se@ZIF-8 nanoparticles showed efficient antibacterial activity as evidenced by low MIC values. Interestingly, these Se@ZIF-8 NPs not only inhibited biofilm formation inS. marcescens,but also effectively eradicated mature biofilms by degrading the eDNA of the EPS layer. It was validated by confocal laser scanning microscopy and SEM analysis. It was observed that Se@ZIF-8 targeted the Quroum Sensing pathway and reduced its associated virulence factors production. This work opens up a different approach of Se@ZIF-8 nanoparticles as novel antibiotics to treat biofilm-associated infections caused byS. marcescensand offer a solution for antimicrobial resistance.

Neutrophil Extracellular Traps-Inspired Bismuth-Based Polypeptide Nanonets for Synergetic Treatment of Bacterial Infections.

Excessive use of antibiotics and the formation of bacterial biofilms can lead to persistent infections caused by drug-resistant bacteria, rendering ineffective immune responses and even life-threatening. There is an urgent need to explore synergistic antibacterial therapies across all stages of infection. Drawing inspiration from the antibacterial properties of neutrophil extracellular traps (NETs) and integrating the bacterial biofilm dispersal mechanism involving boronic acid-catechol interaction, the multifunctional bismuth-based polypeptide nanonets (PLBA-Bi-Fe-TA) are developed. These nanonets are designed to capture bacteria through a coordination complex involving cationic polypeptides (PLBA) with boronic acid-functionalized side chains, alongside metal ions (bismuth (Bi) and iron (Fe)), and tannic acid (TA). Leveraging the nanoconfinement-enhanced high-contact network-driven multiple efficiency, PLBA-Bi-Fe-TA demonstrates the excellent ability to swiftly capture bacteria and their extracellular polysaccharides. This interaction culminates in the formation of a highly hydrophilic complex, effectively enabling the rapid inhibition and dispersion of antibiotic-resistant bacterial biofilms, while Fe-TA shows mild photothermal ability to further assist fluffy mature biofilm. In addition, Bi is beneficial to regulate the polarization of macrophages to pro-inflammatory phenotype to further kill escaping biofilm bacteria. In summary, this novel approach offers a promising bionic optimization strategy for treating bacterial-associated infections at all stages through synergetic treatment.

Influence of carbonyl cyanide m-chlorophenyl hydrazone on biofilm dynamics, protease, and siderophore production by Burkholderia pseudomallei

Efflux pump inhibitors are a potential therapeutic strategy for managing antimicrobial resistance and biofilm formation. This article evaluated the effect of carbonyl cyanide m-chlorophenyl hydrazone (CCCP) on the biofilm growth dynamics and the production of virulence factors by Burkholderia pseudomallei. The effects of CCCP on planktonic, growing, and mature biofilm, interaction with antibacterial drugs, and protease and siderophore production were assessed. CCCP MICs ranged between 128 and 256 µM. The CCCP (128 µM) had a synergic effect with all the antibiotics tested against biofilms. Additionally, CCCP reduced (p < .05) the biomass of biofilm growth and mature biofilms at 128 and 512 µM, respectively. CCCP also decreased (p < .05) protease production by growing (128 µM) and induced (p < .05) siderophore release by planktonic cells (128 µM) growing biofilms (12.8 and 128 µM) and mature biofilms (512 µM). CCCP demonstrates potential as a therapeutic adjuvant for disassembling B. pseudomallei biofilms and enhancing drug penetration.

New insights into the mechanism of azo dye biodegradation by Lactococcus lactis

Azo dye reduction by syntrophic microbial communities is still unclear, with conflicting observations reported in the literature. In this study, the biodegradation mechanism of a model azo dye by Lactococcus lactis strain LLSP-01 isolated from an acidogenic reactor system was investigated. Proteomics and RT-PCR analysis results showed that LLSP-01 azoreductase (AzoL) was not expressed by the isolate during exposure to Direct Black 22. In contrast, the mechanism appears to involve biosorption by glycoconjugates, particularly exopolysaccharides (EPS) and rhamnolipids, as proteins from the LPS O-antigen metabolism were statistically higher expressed in cells challenged with the target compound. Based on the proteomic observations, it is hypothesized that Direct Black 22 is adsorbed into the biofilm matrix and indirectly reduced by an SDR family oxidoreductase, in a mechanism mediated by riboflavin carriers. Induction of a ring-cleaving dioxygenase in the presence of azo dye indicates further degradation of the resulting aromatic amines. The collected results show that azo dye reduction by L. lactis is mediated by enzymes with broad range specificity, and not the typical azoreductases. This information can assist in the design of new strategies for the bioremediation of textile azo dyes.

Biofilm Production and Its Implications in Pediatrics.

Biofilms, aggregates of bacteria enclosed in a self-produced matrix, have been implicated in various pediatric respiratory infections, including acute otitis media (AOM), otitis media with effusion (OME), adenoiditis, protracted bacterial bronchitis, and pulmonary exacerbations in cystic fibrosis. These infections are prevalent in children and often associated with biofilm-producing pathogens, leading to recurrent and chronic conditions. Biofilms reduce antibiotic efficacy, contributing to treatment failure and disease persistence. This narrative review discusses biofilm production by respiratory pathogens such as Streptococcus pneumoniae, non-typeable Haemophilus influenzae, Pseudomonas aeruginosa, and Staphylococcus aureus. It examines their mechanisms of biofilm formation, antibiotic resistance, and the challenges they present in clinical treatment. Various antibiofilm strategies have shown promise in vitro and in animal studies, including the use of N-acetylcysteine, enzymes like dispersin B, and agents disrupting quorum sensing and biofilm matrix components. However, their clinical application, particularly in children, remains limited. Traditional treatments for biofilm-associated diseases have not significantly evolved, even with biofilm detection. The transition from experimental findings to clinical practice is complex and requires robust clinical trials and standardized biofilm detection protocols. Addressing biofilms in pediatric respiratory infections is crucial for improving treatment outcomes and managing recurrent and chronic diseases effectively.

Recent Progress in Multifunctional Stimuli-Responsive Combinational Drug Delivery Systems for the Treatment of Biofilm-Forming Bacterial Infections.

Drug-resistant infectious diseases pose a substantial challenge and threat to medical regimens. While adaptive laboratory evolution provides foresight for encountering such situations, it has inherent limitations. Novel drug delivery systems (DDSs) have garnered attention for overcoming these hurdles. Multi-stimuli responsive DDSs are particularly effective due to their reduced background leakage and targeted drug delivery to specific host sites for pathogen elimination. Bacterial infections create an acidic state in the microenvironment (pH: 5.0-5.5), which differs from normal physiological conditions (pH: 7.4). Infected areas are characterized by the overexpression of hyaluronidase, gelatinase, phospholipase, and other virulence factors. Consequently, several effective stimuli-responsive DDSs have been developed to target bacterial pathogens. Additionally, biofilms, structured communities of bacteria encased in a self-produced polymeric matrix, pose a significant challenge by conferring resistance to conventional antimicrobial treatments. Recent advancements in nano-drug delivery systems (nDDSs) show promise in enhancing antimicrobial efficacy by improving drug absorption and targeting within the biofilm matrix. nDDSs can deliver antimicrobials directly to the biofilm, facilitating more effective eradication of these resilient bacterial communities. Herein, this review examines challenges in DDS development, focusing on enhancing antibacterial activity and eradicating biofilms without adverse effects. Furthermore, advances in immune system modulation and photothermal therapy are discussed as future directions for the treatment of bacterial diseases.

Preparation of porous plate from municipal solid waste incineration fly ash and its application in a biofilm batch reactor

AbstractThe reutilization of municipal solid waste incineration (MSWI) fly ash is a prominent area of research. This study focused on creating fly ash porous plate filler (FAPPF) by using techniques, such as water extraction, milling, component adjustment, and sintering. The produced FAPPF was then used to cultivate a biofilm for wastewater treatment. The key parameters included a two‐stage water extraction process with a 5:1 liquid‐to‐solid ratio; milling for 1, 2, and 4 h; component adjustment using waste glass powder, milled fly ash, palygorskite powder, and peanut shell powder at a 7:1:1:1 mass ratio; and sintering temperatures ranging from 700 to 1000°C. For the biofilm cultivation and treatment, this study employed semisimulated sewage in a sequencing biofilm batch reactor system. The results revealed the FAPPF had no heavy metal leaching, with a porosity of 48.53%–54.68%. Approximately 90% of its composition was derived from waste materials. Furthermore, scanning electron microscopy microanalysis revealed an internally stable liquid‐phase sintering structure. Finally, a mature biofilm developed in 21 days, achieving maximum removal rates of 95.48% for chemical oxygen demand and 78.4% for ammonia nitrogen. This article confirms the sustainable recycling potential of MSWI fly ash.

Activity of Synthetic Peptide KP and Its Derivatives against Biofilm-Producing Escherichia coli Strains Resistant to Cephalosporins.

Bacterial resistance to β-lactam antibiotics, particularly new generation cephalosporins, is a major public health concern. In Escherichia coli, resistance to these antibiotics is mainly mediated by extended-spectrum β-lactamases (ESBL), which complicates a range of health-threatening infections. These infections may also be biofilm-related, making them more difficult to treat because of the higher tolerance to conventional antibiotics and the host immune response. In this study, we tested as potential new drug candidates against biofilm-forming ESBL-producing E. coli four antimicrobial peptides previously shown to have antifungal properties. The peptides proved to be active in vitro at micromolar concentrations against both sensitive and ESBL-producing E. coli strains, effectively killing planktonic cells and inhibiting biofilm formation. Quantitative fluorescence intensity analysis of three-dimensional reconstructed confocal laser scanning microscopy (CLSM) images of mature biofilm treated with the most active peptide showed significant eradication and a reduction in viable bacteria, while scanning electron microscopy (SEM) revealed gross morphological alterations in treated bacteria. The screening of the investigated peptides for antibacterial and antibiofilm activity led to the selection of a leading candidate to be further studied for developing new antimicrobial drugs as an alternative treatment against microbial infections, primarily associated with biofilms.

Cutibacterium acnes biofilm formation is influenced by bone microenvironment, implant surfaces and bacterial internalization

BackgroundThe bacterial persistence, responsible for therapeutic failures, can arise from the biofilm formation, which possesses a high tolerance to antibiotics. This threat often occurs when a bone and joint infection is diagnosed after a prosthesis implantation. Understanding the biofilm mechanism is pivotal to enhance prosthesis joint infection (PJI) treatment and prevention. However, little is known on the characteristics of Cutibacterium acnes biofilm formation, whereas this species is frequently involved in prosthesis infections.MethodsIn this study, we compared the biofilm formation of C. acnes PJI-related strains and non-PJI-related strains on plastic support and textured titanium alloy by (i) counting adherent and viable bacteria, (ii) confocal scanning electronic microscopy observations after biofilm matrix labeling and (iii) RT-qPCR experiments.ResultsWe highlighted material- and strain-dependent modifications of C. acnes biofilm. Non-PJI-related strains formed aggregates on both types of support but with different matrix compositions. While the proportion of polysaccharides signal was higher on plastic, the proportions of polysaccharides and proteins signals were more similar on titanium. The changes in biofilm composition for PJI-related strains was less noticeable. For all tested strains, biofilm formation-related genes were more expressed in biofilm formed on plastic that one formed on titanium. Moreover, the impact of C. acnes internalization in osteoblasts prior to biofilm development was also investigated. After internalization, one of the non-PJI-related strains biofilm characteristics were affected: (i) a lower quantity of adhered bacteria (80.3-fold decrease), (ii) an increase of polysaccharides signal in biofilm and (iii) an activation of biofilm gene expressions on textured titanium disk.ConclusionTaken together, these results evidenced the versatility of C. acnes biofilm, depending on the support used, the bone environment and the strain.

Trash into Treasure: Nano-coating of Catheter Utilizes Urine to Deprive H2S Against Persister and Rip Biofilm Matrix.

Bacteria-derived hydrogen sulfide (H2S) often contributes to the emergence of antibiotic-recalcitrant bacteria, especially persister (a sub-population of dormant bacteria), thus causing the treatment failure of Catheter-associated urinary tract infection (CAUTI). Here, an H2S harvester nanosystem to prevent the generation of persister bacteria and disrupt the dense biofilm matrix by the self-adaptive ability of shape-morphing is prepared. The nanosystem possesses a core-shell structure that is composed of liquid metal nanoparticle (LM NP), AgNPs, and immobilized urease. The nanosystem decomposes urea contained in urine to generate ammonia for eliminating bacteria-derived H2S. Depending on the oxidative layer of liquid metal, the nanosystem also constitutes a long-lasting reservoir for temporarily storing bacteria-derived H2S, when urease transiently overloads or in the absence of urine in a catheter. Depriving H2S can prevent the emergence of persistent bacteria, enhancing the bacteria-killing efficiency of Ga3+ and Ag+ ions. Even when the biofilm has formed, the urine flow provides heat to trigger shape morphing of the LM NP, tearing the biofilm matrix. Collectively, this strategy can turn trash (urea) into treasure (H2S scavengers and biofilm rippers), and provides a new direction for the antibacterial materials application in the medical field.

The influence of different methods of decontamination of microbial biofilms formed on eggshells

According to "food legislation" requirements, all eggs entering the production of egg products must be disinfected. Therefore, developing technologies for decontaminating chicken eggs before use for food purposes is a promising work direction in chicken egg production and storage. This research aimed to identify the microbiota of chicken eggs with varying degrees of shell contamination and determine the influence of different methods of decontaminating microbial biofilms formed on eggshells. It was set up that the quantitative content of microorganisms on the surface of chicken eggs ranged from 103 CFU to 106 CFU/ml of washing and depended on the contamination of the shell with droppings. Lactobacillus spp., Bacillus spp., Corynebacterium, Staphylococcus were among the genera of bacteria that prevailed on the clean chicken shell, which were isolated in 30-50% of cases, and gram-negative microbiota was practically absent. The constant release of gram-positive bacteria is noted on the contaminated eggshell, and the frequency of identification of gram-negative microbiota of the Enterobacteriaceae genus and non-fermenting genera Pseudomonas and Psychrobacter increases. That is, the microbial scape of the microbiota of the chicken shell depends on its cleanliness, and the presence of a dirty surface increases the frequency of allocation of the resident microflora of the gastrointestinal tract. It was found that the working solution of the disinfectant Vircon S destroyed planktonic bacteria applied to the eggshell in an average of 2 minutes of exposure, stabilised water ozone for 1 minute, gaseous ozone for 3 minutes, and the action of ultraviolet rays with a length of 253.7 nm for 25-30 min. At the same time, using these disinfection methods on bacteria formed in a biofilm on the eggshell did not cause a bactericidal action during this time. To significantly reduce bacteria in the biofilm using these methods, it is necessary to increase the exposure time of the biocide by 2-3 times. Therefore, the complex structure of the eggshell and the multi-layered matrix of biofilms provide better protection for bacteria against the influence of the investigated disinfection methods.

Effect of Novel and Traditional Intracanal Medicaments on Biofilm Viability and Composition

IntroductionThe aim of this study was to test the hypothesis that a combination of D-amino acids (DAAs) and trans-cinnamaldehyde (TC) demonstrates superior antibiofilm activity to calcium hydroxide (CH) and untreated controls. MethodsIn this 3-part in vitro study, the concentration of DAAs (D-methionine, D-leucine, D-tyrosine, and D-tryptophan) that would significantly decrease Enterococcus faecalis and Actinomyces naeslundii biofilm biomass was first determined. Then, the effect of TC + selected DAAs on polymicrobial biofilms was characterized by quantifying the biomass and biofilm viability. Finally, the antibiofilm effects of TC + DAA was compared with CH and untreated controls by (i) determining bacterial viability and (ii) quantifying biofilm matrix composition using selective fluorescence-binding analysis. Statistical analysis was performed using one-way ANOVA and appropriate multiple comparisons test, with P < .05 considered as statistically significant. ResultsTC (0.06%) + D-tyrosine (1 mM) + D-tryptophan (25 mM) significantly reduced the biomass and biofilm viability compared to the control (P < .05). While no significant difference was observed between TC + DAA and CH in the cultivable bacterial counts (P > .05), confocal microscopy demonstrated a significantly greater percentage of dead bacteria in TC + DAA-treated biofilms compared to CH and the control (P < .05). TC + DAA significantly decreased the biovolume and all the examined components of the biofilm matrix quantity compared to the control, while CH significantly reduced only the exopolysaccharide quantity (P < .05). ConclusionThe combination of TC + D-tyrosine + D-tryptophan demonstrated superior antibiofilm activity (biofilm bacterial killing and reduction of matrix quantity) to CH and has potential to be developed as an intracanal medicament.

Stimuli-Responsive Release-Active Dressing: A Promising Solution for Eradicating Biofilm-Mediated Wound Infections.

Biofilm-mediated wound infections pose a significant challenge due to the limitations of conventional antibiotics, which often exhibit narrow-spectrum activity, fail to eliminate recurrent bacterial contamination, and are unable to penetrate the biofilm matrix. While the search for alternatives has explored the use of metal nanoparticles and synthetic biocides, these solutions often suffer from unintended toxicity to surrounding tissues and lack controlled administration and release. In this study, we engineered a pH-responsive release-active dressing film based on carboxymethyl cellulose, incorporating a synthetic antibacterial molecule (SAM-17). The dressing film exhibited optimal mechanical stability for easy application and demonstrated excellent fluid absorption properties, allowing for prolonged moisturization at the site of injury. The film exhibited pH-dependent release of cargo, with 78% release within 24 h at acidic pH, enabling targeted antibacterial drug delivery within the wound microenvironment. Furthermore, the release-active film effectively eliminated repeated challenges of bacterial contamination. Remarkably, the film demonstrated a minimal toxicity profile in both in vitro and in vivo models. The film eliminated preformed bacterial biofilms, achieving a reduction of 2.5 log against methicillin-resistant Staphylococcus aureus (MRSA) and 4.1 log against vancomycin-resistant S. aureus (VRSA). In a biofilm-mediated MRSA wound infection model, this release-active film eradicated the biofilm-embedded bacteria by over 99%, resulting in accelerated wound healing. These findings highlight the potential of this film as an effective candidate for tackling biofilm-associated wound infections.

Previously unknown regulatory role of extracellular RNA on bacterial directional migration.

Bacterial directional migration plays a significant role in bacterial adaptation. However, the regulation of this process, particularly in young biofilms, remains unclear. Here, we demonstrated the critical role of extracellular RNA as part of the Universal Receptive System in bacterial directional migration using a multidisciplinary approach, including bacterial culture, biochemistry, and genetics. We found that the destruction or inactivation of extracellular RNA with RNase or RNA-specific antibodies in the presence of the chemoattractant triggered the formation of bacterial "runner cells» in what we call a "panic state" capable of directional migration. These cells quickly migrated even on the surface of 1.5% agar and formed evolved colonies that were transcriptionally and biochemically different from the ancestral cells. We have also shown that cell-free DNA from blood plasma can act as a potent bacterial chemoattractant. Our data revealed a previously unknown role of bacterial extracellular RNA in the regulation of bacterial migration and have shown that its destruction or inhibition triggered the directional migration of developing and mature biofilms towards the chemoattractant.