Bacterial Biofilm Formation Research Articles

Designing Impact Resistance and Robustness into Slippery Lubricant Infused Porous Surfaces.

Slippery lubricant infused porous surfaces (SLIPS) have the potential to address daunting challenges such as undesirable surface fouling/biofouling, icing, etc. However, the depletion of lubricants hampers their practical utility. As a solution, here a rational strategy is introduced that operates synergistically in three parts. First, ultra-high capillary pressure is exploited from the reticular structure of metal-organic frameworks (MOFs) with sub-nanometer pores. Second, the need for geometric compatibility is demonstrated; the lubricant chain diameter must be smaller than the MOF pore to enable lubricant chain intercalation. Lastly, the MOF pore chemistry is tailored to achieve a strong intermolecular interaction for any given MOF/lubricant combination. The strategy is investigated through experiments and quantum/molecular simulations, which show that the approach helps lock the intercalated lubricant chains inside the MOF pores and forms a non-conventional supramolecular structure. The resulting SLIPS (including those with fluorine-free chemistry) not only show typical low wetting hysteresis, friction, and ice adhesion but are uniquely resistant to more stringent tests such as continuous dripping and sliding of water droplets (up to 50hrs), repeated impacts of high-speed water jets (liquid impact Weber number >4×104) and prevent bacterial biofilm formation even in dynamic flow conditions. The findings may widen the practical applications of SLIPS.

Synergistic effect of antibiotics, α-linolenic acid and solvent type against Staphylococcus aureus biofilm formation.

A promising approach to the treatment of bacterial infections involves inhibiting the quorum sensing (QS) mechanism to prevent the formation and growth of bacterial biofilm. While antibiotics are used to kill remaining bacteria, QS inhibitors (QSIs) allow for antibiotic doses to be reduced. This study focuses on evaluating the synergy between gentamicin sulphate (GEN), tobramycin (TOB), or azithromycin (AZM) with linolenic acid (LNA) against the formation of an early Staphylococcus aureus biofilm. Minimum biofilm inhibitory concentration (MBIC) was determined using the resazurin reduction assay for all antibiotics and LNA. The reduction of biofilm mass was assessed using the crystal violet (CV) assay. We have also evaluated the effect of dimethyl sulfoxide with TWEEN (DMSO_T) on early biofilm formation. Synergy was determined by metabolic activity assay and fractional biofilm inhibitory concentration (FBIC). DMSO_T at a concentration of 1% enhanced early biofilm formation, but also decreased the doses of antibiotic needed to reduce the biofilm by up to 8 times. Adding LNA at a concentration of 32µg/ml or 64µg/ml allowed up to a 32-fold reduction of antibiotic doses for GEN and TOB and a 4-fold reduction for AZM. LNA's use in combination with various antibiotics could reduce their doses and help fight drug-resistant bacteria in the biofilm.

Impact of Biofilms on Surface Properties of Polymethyl Methacrylate (PMMA) Resins.

Poly(methyl methacrylate) (PMMA) resins are widely used in medical and dental applications. Their susceptibility to bacterial biofilm formation poses significant challenges related to material degradation and infection risk. This study investigated the effects of Pseudomonas aeruginosa (P. aeruginosa) and Staphylococcus aureus (S. aureus) biofilms on PMMA resin surface properties over a 45-day period at 35°C. The study examined various parameters including biofilm adhesion, morphology, surface roughness, hydrophobicity, solid fraction, and zeta potential. PMMA resin specimens were inoculated with bacteria and incubated for 45 days. Biofilm adhesion was visually assessed, while surface characterization was conducted using scanning electron microscopy (SEM), atomic force microscopy (AFM), roughness analysis, contact angle measurements, solid fraction determination, and zeta potential analysis. The P. aeruginosa and S. aureus isolates were selected based on their biofilm-positive characteristics, which were further confirmed using Congo red and biofilm formation assays through crystal violet staining and spectrophotometric analysis. The results demonstrated robust biofilm adhesion on PMMA surfaces. SEM and AFM imaging revealed textured surfaces with elevated structures and depressions within the biofilm matrix. Biofilm-exposed resins exhibited significantly increased roughness (Ra = 164.5 nm, Rq = 169.5 nm) and hydrophobicity (mean angle = 85.5°-90.5°) compared to control samples (Ra = 38-50 nm, angle = 55°). Solid fraction measurements indicated a denser biofilm matrix on exposed resins (0.908) compared to controls (0.65). Additionally, zeta potential values were more negative for biofilm-exposed resins (mean = -84.2 mV) than controls (-45.0 mV). These findings underscore the substantial alterations in PMMA resin surface properties induced by bacterial biofilms, emphasizing the critical need for strategies to prevent biofilm formation and mitigate associated risks in healthcare settings. Future research should focus on developing anti-biofilm coatings or treatments to preserve the integrity and functionality of PMMA materials.

A bifunctional nanocomplex with remineralizing and antibacterial activities to interrupt dental caries

Dental caries result from an imbalance between the demineralization and remineralization of dental hard tissues, primarily caused by biofilm accumulation. According to the theory of interrupting dental caries, effective anticaries materials and techniques should possess both remineralizing and antibacterial properties. However, current anticaries materials cannot replicate amelogenesis, which is crucial for remineralization and for preventing bacterial adhesion and biofilm formation. In this study, silk fibroin (SF) loaded with benzalkonium chloride (BZC) successfully formed an SF-BZC composite. This composite stabilized amorphous calcium phosphate (ACP), creating an ACP@SF-BZC dual-functional nanocomplex with both remineralizing and antibacterial properties. ACP@SF-BZC demonstrated significant anti-adhesion and biofilm inhibitory effects against Streptococcus mutans and Streptococcus sobrinus. Moreover, compared to fluoride, ACP@SF-BZC significantly enhanced the remineralization of demineralized enamel surfaces, forming a stable remineralized layer with improved mechanical properties, both in vitro and in vivo. In summary, the dual-function ACP@SF-BZC nanocomplex, with its remineralizing and antibacterial effects, offers a promising alternative for preventing and arresting enamel caries.

Evaluation of bacterial anti-adhesion and sustained antimicrobial abilities of chlorhexidine gluconate-loaded phase-transited lysozyme nanoparticle suspension on dentine: An exvivo study.

To evaluate the effect of chlorhexidine gluconate-loaded phase-transited lysozyme (CHG@PTL) coating on inhibiting bacterial adhesion and biofilm formation in an exvivo root canal dentine model. The physicochemical and structural characteristics of CHG@PTL nanoparticle suspension and its coating formed on the dentine surface were analysed by thioflavin T fluorescence assay, transmission electron microscopy and confocal laser scanning microscopy (CLSM). The sustained chlorhexidine release profile of the CHG@PTL coating on the dentine surface was compared with that of the 2% CHG solution. By comparing with phosphate-buffered saline, 1% sodium hypochlorite and 2% CHG solutions, the sustained antibacterial ability of the CHG@PTL coating and its effects on adhesion and biofilm formation of three types of bacteria (E. faecalis, S. mutans, and A. viscous) were analysed in exvivo root canal dentine models using the serial plate transfer test (SPTT) and CLSM with live/dead bacterial staining, respectively. CHG promoted the lysozyme protein to form a higher proportion of β-sheet structure during phase transition. In the CHG@PTL nanoparticle suspension, characteristic drug-loaded nanospheres with a high concentration of CHG molecules inside and an outer PTL nanofilm were observed, and they formed a thinner and tighter coating on the dentine surface. The CHG@PTL coating on the dentine surface showed a significantly higher cumulative release amount of chlorhexidine than that of 2% CHG (p < .05). The results of SPTT showed that the CHG@PTL coating had a longer antibacterial duration than the control groups. After 12 h of incubation, a higher number of bacteria were agglutinated on the CHG@PTL coating surface compared to the control groups (p < .05). After 7 days of incubation, the number of agglutinated bacteria significantly decreased. At two time points, the percentage of dead bacteria on the CHG@PTL coating surface was the highest among all experimental groups based on CLSM observation (over 99.9% for all three bacteria, p < .001). CHG@PTL nanoparticle suspension could form an antimicrobial coating on the surface of dentine with a novel 'agglutinating bacteria and sterilizing' mode.

Multifunctional Carbon Dots Derived from Human Hair for Fast Healing Wounds Together with Oleogels

In the complex process of wound healing, oleogels (OG) are suitable as the primary component of dressing materials, but they cannot meet the diverse requirements at different healing stages. In this study, a new kind of carbon dots (CrCi‐CDs) prepared by carbonizing human hair, exhibits excellent hemostatic, antibacterial, anti‐inflammatory, and pro‐angiogenic properties and thus supports the various stages of wound healing effectively. Such CrCi‐CDs are incorporated into OG to produce a CrCi‐CD/OG composite material with enhanced multifunctional capabilities, significantly outperforming OG alone. Various experiments in vitro and in vivo confirm that the CrCi‐CDs/OG can rapidly achieve hemostasis at the initial stage of wound formation, subsequently inhibit bacterial proliferation and biofilm formation, improve the complex microenvironment surrounding the wound, and promote neovascularization, ultimately accelerating the wound repair.

Dual-action defense: A photothermal and controlled nitric oxide-releasing coating for preventing biofilm formation

Biofilms formed by pathogenic bacteria on biomedical devices and implants pose a considerable challenge due to their resistance to conventional treatments and their role in severe infections. Preventing biofilm formation is strategically more advantageous than attempting to eliminate the mature biofilms, particularly in addressing the persistence of such formations. In this context, a dual-action antibiofilm coating is developed, utilizing S-nitrosothiols functionalized candle soot (CS), which capitalizes on CS’s strong light absorption for photothermal therapy and the controlled release of nitric oxide (NO) from S-nitrosothiols to inhibit biofilm formation. This coating exhibits stable and efficient light-to-heat conversion, along with the ability to release NO gradually at physiological temperatures and to rapidly release NO on demand when triggered by a near-infrared (NIR) laser. Under NIR irradition, the coating generates heat swiftly and releases substantial amounts of NO, which synergistically disrupts bacterial membranes, leading to the leakage of intracellular components and the effective eradication of surface-adhered bacteria. In the absence of NIR irradiation, the coating continuously releases low concentrations of NO, which depletes exopolysaccharides and impedes biofilm formation. The antibiofilm efficacy of this coating is assessed against Staphylococcus aureus and Pseudomonas aeruginosa, demonstrating marked reductions in bacterial viability and biofilm formation in vitro. Additionally, the coating exhibits minimal cytotoxicity and can be easily applied to diverse substrates. This study underscores the potential of this coating as a broad-spectrum, non-toxic approach for preventing biofilm-related complications in biomedical applications.

Iron Single-Atom Nanozyme with Inflammation-Suppressing for Inhibiting Multidrug-Resistant Bacterial Infection and Facilitating Wound Healing.

Infection with drug-resistant bacteria and the formation of biofilms are the main factors contributing to wound healing insufficiency. Antibacterial agents with enzyme-like properties have exhibited considerable potential for efficient eradication of drug-resistant microorganisms due to their superior sensitivities and minimal side effects. In this work, we prepared a kind of Fe-centered single-atom nanozyme (Fe-SAzyme) with high biocompatibility and stability via a facile one-pot hydrothermal method, which was suitable for the treatment of wounds infected with drug-resistant bacteria. The Fe-SAzyme exhibited remarkable peroxidase-like catalytic activities, catalyzing the conversion of hydrogen peroxide (H2O2) to highly toxic hydroxyl radicals (•OH), which could not only damage bacterial cells but also inhibit, disrupt, and eradicate the formation of bacterial biofilms. Thus, Fe-SAzyme demonstrated a broad-spectrum antibacterial performance capable of effectively eliminating multidrug-resistant bacteria. The coexistence of ferrous (Fe2+) and ferric (Fe3+) ions in Fe-SAzyme conferred the nanozyme with anti-inflammatory activity, effectively suppressing excessive inflammation. Meanwhile, Fe-SAzyme could significantly downregulate inflammatory cytokines tumor necrosis factor-α and interleukin-1β and upregulate growth factors VEGF and epidermal growth factor, which can prevent bacterial infection, mitigate inflammation, promote fibroblast proliferation, and improve wound closure. Thus, Fe-SAzyme had shown favorable therapeutic efficiency in promoting bacteria-infected wound healing. This study provides Fe-SAzyme as a promising candidate for the development of new strategies to treat multidrug-resistant bacterial infections.

Novel compound, pleuropyronine, and other polyketides isolated from the edible mushroom Pleurotus ostreatus suppress bacterial biofilm formation.

An increase in the number of drug-resistant microbes is a major threat to human health. Bacterial drug resistance is mostly mediated by biofilm formation. In this study, the culture filtrate from the edible mushroom, Pleurotus ostreatus, was fractionated to isolate compounds that inhibit the biofilm formation of six pathogenic bacteria. Notably, we isolated compounds 16 using bioassay-guided chromatographic separations. Spectroscopic and X-ray diffraction analyses identified 1 as a novel fused bicyclic pyrone-furan, named pleuropyronine, whereas 2-6 were known polyketides. Pleuropyronine inhibited biofilm formation in four gram-negative bacteria, with IC50 values ranging from 5.4 to 8.7 µg/mL, whereas 2-6 exhibited IC50 values between 1.0 and 5.3 µg/mL against five bacteria. Additionally, pleuropyronine bioactivity was confirmed by the inhibition of exopolysaccharide and biofilm formation induced by C6-homoserine lactone. Thus, this may serve as a pioneering study on the pharmacological potential of isolated compounds, offering valuable insights for future research.

D-[5-11C]-Glutamine Positron Emission Tomography Imaging for Diagnosis and Therapeutic Monitoring of Orthopedic Implant Infections.

Orthopedic implant infections (OIIs) present diagnostic and therapeutic challenges, owing to the lack of methods to distinguish between active infection and sterile inflammation. To address this unmet need, d-amino-acid-based radiotracers with unique metabolic profiles in microorganisms have emerged as a novel class of infection-specific imaging agents. Given the pivotal role of d-glutamine in bacterial biofilm formation and virulence, herein, we explored the potential of positron emission tomography (PET) imaging with d-[5-11C]-Glutamine (d-[5-11C]-Gln) for early detection and treatment monitoring of OIIs. In vitro studies confirmed an active uptake of d-[5-11C]-Gln by Staphylococcus aureus (S. aureus) biofilm commonly associated with OIIs. In vivo evaluations included PET imaging comparisons with d-[5-11C]-Gln vs l-[5-11C]-Gln or 2-deoxy-2-[18F]-fluoroglucose ([18F]-FDG) in a rat OII model with tibial implantation of sterile or S. aureus-colonized stainless-steel screws before and after treatment. These studies demonstrated that the uptake of d-[5-11C]-Gln was significantly higher in the infected screws than that in sterile screws (∼3.4-fold, p = 0.008), which displayed significantly higher infection-to-background muscle uptake ratios (∼2-fold, p = 0.011) with d-[5-11C]-Gln as compared to l-[5-11C]-Gln. Following a 3 week vancomycin treatment, imaging with d-[5-11C]-Gln showed a significant reduction in uptake at the infected sites (∼3-fold, p = 0.0008). Further regression analyses revealed a superior correlation of residual infection-associated radiotracer uptake with the postimaging ex vivo bacterial counts for d-[5-11C]-Gln (k = 0.473, R2 = 0.796) vs [18F]-FDG (k = 0.212, R2 = 0.434), suggesting that d-[5-11C]-Gln PET had higher sensitivity for detecting residual bacterial burden than [18F]-FDG PET. Our results demonstrate the translational potential of d-[5-11C]-Gln PET imaging for noninvasive detection and treatment monitoring of OIIs.

Holliday junction resolvase RuvC targets biofilm eDNA and confers plant resistance to vascular pathogens.

A biofilm lifestyle is critical for bacterial pathogens to colonize and protect themselves from host immunity and antimicrobial chemicals in plants and animals. The formation and regulation mechanisms of phytobacterial biofilm are still obscure. Here we found that the protein Ralstonia solanacearum resistance to ultraviolet C (RuvC) is highly abundant in biofilm and positively regulates pathogenicity by controlling systemic movement in tomato xylem. RuvC protein accumulates at the later stage of biofilm development and specifically targets Holliday junction (HJ)-like structures to disrupt the biofilm extracellular DNA (eDNA) lattice, thus facilitating biofilm dispersal. Recombinant RuvC protein can resolve extracellular HJ to prevent bacterial biofilm formation. Heterologous expression of R. solanacearum or Xanthomonas oryzae pv. oryzae RuvC with plant secretion signal in tomato or rice confers resistance to bacterial wilt or bacterial blight disease, respectively. Plant chloroplast-localized HJ resolvase monokaryotic chloroplast 1 (MOC1), which shares structural similarity with bacterial RuvC, shows a strong inhibitory effect on bacterial biofilm formation. Relocalization of SlMOC1 to apoplast in tomato roots leads to increased resistance to bacterial wilt. Our novel finding reveals a critical pathogenesis mechanism of R. solanacearum and provides an efficient biotechnology strategy to improve plant resistance to bacterial vascular disease.

Gum Arabic as a potential candidate in quorum quenching and treatment of periodontal diseases.

Periodontal diseases are chronic inflammatory conditions influenced by bacterial biofilm formation and host immune responses, affecting millions worldwide. Traditional treatments like mechanical debridement and systemic antibiotics often face limitations, including biofilm resilience and antibiotic resistance. Gum Arabic (GA), a natural exudate from Acacia trees, presents a promising alternative with its anti-biofilm and anti-inflammatory properties. This review highlights the role of GA in periodontal therapy, particularly its ability to interfere with quorum sensing (QS) pathways, specifically the AI-2 signaling system used by key periodontal pathogens such as Porphyromonas gingivalis, Aggregatibacter actinomycetemcomitans, and Fusobacterium nucleatum. By disrupting QS, GA inhibits biofilm formation, reduces bacterial virulence, and promotes a balanced oral microbiome. GA's prebiotic properties also encourage the growth of beneficial bacteria, enhancing the host's immune response while preserving the systemic microbiome. Clinical studies demonstrate GA's effectiveness as an adjunct in periodontal therapy, with significant reductions in plaque accumulation, gingival inflammation, and bleeding. This highlights GA's potential as a natural therapeutic agent, offering an effective, antibiotic-sparing option in managing periodontal disease. However, further research is warranted to fully establish GA's role in comprehensive periodontal care and its long-term benefits.

Engineering the novel azobenzene-based molecular photoswitches for suppressing bacterial infection through dynamic regulation of biofilm formation.

Bacterial biofilm is a strong fortress for bacteria to resist harsh external environments, which can enhance their tolerance and exacerbate the drug/pesticide resistance risk. Currently, photopharmacology provides an advanced approach via precise spatiotemporal control for regulating biological activities by light-controlling the molecular configurations, thereby having enormous potential in the development of drug/pesticides. To further expand the photopharmacology application for discovering new antibiofilm agents, we prepared a series of light-controlled azo-active molecules and explored their photo isomerization, fatigue resistance, and anti-biofilm performance. Furthermore, their mechanisms of inhibiting biofilm formation were systematically investigated. Overall, designed azo-derivative A11 featured excellent anti-Xoo activity with an half-maximal effective concentration (EC50) value of 5.45 μg mL-1, and the EC50 value could be further elevated to 2.19 μg mL-1 after ultraviolet irradiation (converted as cis-configuration). The photo-switching behavior showed that A11 had outstanding anti-fatigue properties. An in-depth analysis of the action mechanism showed that A11 could effectively inhibit biofilm formation and the expression of relevant virulence factors. This performance could be dynamically regulated via loading with private light-switch property. In this work, designed light-controlled azo molecules provide a new model for resisting bacterial infection via dynamic regulation of bacterial biofilm formation. © 2024 Society of Chemical Industry.

Synergistic effects of thermally reduced graphene oxide/zinc oxide composite material on microbial infection for wound healing applications

Infections originating from pathogenic microorganisms can significantly impede the natural wound-healing process. To address this obstacle, innovative bio-active nanomaterials have been developed to enhance antibacterial capabilities. This study focuses on the preparation of nanocomposites from thermally reduced graphene oxide and zinc oxide (TRGO/ZnO). The hydrothermal method was employed to synthesize these nanocomposites, and their physicochemical properties were comprehensively characterized using X-ray diffraction analysis (XRD), High-resolution transmission electron microscopy (HR-TEM), Fourier-transform infrared (FT-IR), Raman spectroscopy, UV-vis, and field-emission scanning electron microscopy (FE-SEM) techniques. Subsequently, the potential of TRGO/ZnO nanocomposites as bio-active materials against wound infection-causing bacteria, including Staphylococcus aureus, Pseudomonas aeruginosa, and Escherichia coli, was evaluated. Furthermore, the investigated samples show disrupted bacterial biofilm formation. A reactive oxygen species (ROS) assay was conducted to investigate the mechanism of nanocomposite inhibition against bacteria and for further in-vivo determination of antimicrobial activity. The MTT assay was performed to ensure the safety and biocompatibility of nanocomposite. The results suggest that TRGO/ZnO nanocomposites have the potential to serve as effective bio-active nanomaterials for combating pathogenic microorganisms present in wounds.

Co-selective effect of dissolved organic matter and chlorine on the bacterial community and their antibiotic resistance in biofilm of drinking water distribution pipes

The proliferation of pathogenic bacteria and antibiotic resistance genes (ARGs) in the biofilm of drinking water distribution pipes poses a serious threat to human health. This work adopted 15 polyethylene (PE) pipes to study the co-selective effect of dissolved organic matter (DOM) and chlorine on the bacterial community and their antibiotic resistance in biofilm. The results indicated that ozone and granular activated carbon (O3-GAC) filtration effectively removed lignins and proteins from DOM, and chlorine disinfection eliminated carbohydrate and unsaturated hydrocarbons, which both contributed to the inhibition of bacterial growth and biofilm formation. After O3-GAC and disinfection treatment, Porphyrobacter, unclassified_d_bacteria, and Sphingopyxis dominated in the biofilm bacterial community. Correspondingly, the bacterial metabolism pathways, including the phosphotransferase system, phenylalanine, tyrosine and tryptophan biosynthesis, ABC transporters, and starch and sucrose metabolism, were downregulated significantly (p < 0.05), compared to the sand filtration treatment. Under such a situation, extracellular polymeric substances (EPS) secretion was inhibited in biofilm after O3-GAC and disinfection treatment, postponing the interaction between EPS protein and pipe surface, preventing bacteria, especially pathogens, from adhering to the pipe surface to form biofilm, and restraining the spread of ARGs. This study revealed the effects of various water filtration and disinfection processes on bacterial growth, metabolism, and biofilm formation on a molecular level, and validated that the O3-GAC filtration followed by chlorine disinfection is an effective and promising pathway to control the microbial risk of drinking water.

Nicotine promotes pathogenic bacterial growth and biofilm formation in peri-implant.

Introduction. Peri-implantitis is a plaque-associated disease that leads to implant loss and arises from bacterial biofilms on the surface of the implant. Smoking is a risk factor for peri-implantitis and impedes treatment effectiveness. Additionally, aryl hydrocarbon receptor (AHR), IL-6, and IL-22 levels are related to peri-implantitis.Aim. We aimed to investigate the effects of nicotine on inflammatory response, bacterial growth and biofilm formation.Hypothesis/Gap Statement. We hypothesized that nicotine promoted pathogenic bacterial growth and biofilm formation, thereby aggravating inflammation.Methodology. The expression of AHR, IL-6 and IL-22 was measured in peri-implant sulci fluid using quantitative PCR and Western blot analyses. The cementum was incubated with bacterial suspension including Porphyromonas gingivalis, Streptococcus sanguinis and Fusobacterium nucleatum and treated with 100, 200, 250 and 300 µg ml-1 nicotine, and then, the absorbance and number of colony-forming units were detected. Biofilm formation was evaluated using the tissue culture plate method and safranin O staining. Carbohydrates and proteins were measured by the phenol-sulfuric acid method and the bicinchoninic acid method, respectively.Results. The results indicated that smoking increased the levels of AHR, IL-6 and IL-22. Functionally, nicotine promoted the growth of P. gingivalis, S. sanguinis and F. nucleatum. Additionally, it promoted the biofilm formation of these bacteria and increased the contents of carbohydrates and proteins.Conclusion. Nicotine promoted bacterial growth and biofilm build-up, suggesting that smoking may aggravate the progression of peri-implantitis.

Synthesis, Antimicrobial and Antibiofilm Activities of Silver(I) Complexes with N-Alkylbenzimidazole Derivatives and Their Molecular Docking Study

This study investigates the antimicrobial and antibiofilm potential of three silver(I) complexes derived from N-alkylbenzimidazole derivatives: bis[(Z)-1-styryl-benzimidazole]silver(I) nitrate (4), bis[(E)-1-styryl-benzimidazole]silver(I) nitrate (5), and bis(1-cinnamyl-benzimidazole)silver(I) nitrate (6). Detailed synthesis and characterization of these complexes are followed by assessments of their efficacy against Staphylococcus aureus, Enterococcus faecalis, Pseudomonas aeruginosa, Escherichia coli, and Candida albicans. Among the tested compounds, complex 6 exhibited the highest antimicrobial activity, with a minimal inhibitory concentration of 6 µmol/L against P. aeruginosa, and significant antibiofilm activity, achieving 67.72% inhibition at a concentration of 3 µmol/L. These interesting findings led us to conduct a molecular docking study to understand the mechanisms of action by investigating the interactions between the silver(I) complexes and some key protein targets involved in bacterial and fungal biofilm formation, including (p)ppGpp synthetase/hydrolases, FtsZ proteins, and pyruvate kinases. This comprehensive approach, combining experimental and computational analyses derived from N-alkylbenzimidazole derivatives, particularly complex 6, which exhibits remarkable efficacy against various pathogens, reveals promising therapeutic applications of these silver(I) complexes and advances our understanding of their potential mode of action against biofilm-associated infections.

Identification of a cellular role of hemolysin co-regulatory protein (Hcp) in Vibrio alginolyticus modulating substrate metabolism and biofilm formation by cAMP-CRP

Cyclic AMP (cAMP) and cAMP receptor protein (CRP) system controls catabolic enzyme expression based on metabolite concentrations in bacteria. Hemolysin co-regulatory protein (Hcp) is well known as a molecular chaperone for virulence factor secretion of the type VI secretion system (T6SS). However, the intracellular role of Hcp involving in bacterial physiological processes remains unknown. To clarify that, we constructed a single hcp mutant strain and analyzed their effects on the physiological processes of Vibrio alginolyticus. The omics results revealed the extensive involvement of Hcp in the catabolic metabolism in bacteria. Simultaneously, Hcp1 and Hcp2 played opposing regulatory roles on the bacterial growth, biofilm formation, and intracellular cAMP-CRP levels during cultivation in a glucose medium. Furthermore, the interacting protein screening and co-immunoprecipitation (Co-IP) assays confirmed that the glucose-specific phosphoenolpyruvate (PEP)-phosphotransferase system (PTS) enzyme IIA component (EIIAglc) was a key interacting partner with Hcp proteins as well as class I adenylyl cyclase (AC-I) in Vibrio alginolyticus. These results indicated that, to achieve cellular homeostasis, Hcp1 and Hcp2 might exert antagonistic and synergistic effects, respectively, on the interaction between EIIAglc and AC thus cooperatively regulating intracellular cAMP-CRP production.

Development of erythromycin loaded PLGA nanoparticles for improved drug efficacy and sustained release against bacterial infections and biofilm formation

Bacterial infections are a common cause of sepsis, often leading to high patient mortality. Such infections are challenging to treat due to bacterial resistance to many existing drugs. Erythromycin (Ery) is a macrolide antibiotic used against bacterial infections with reported resistance. Recently, synthetic poly-lactide co-glycolic acid (PLGA) polymer nanoparticles (NPs) have displayed improved drug delivery characteristics and biocompatibility. In this study, PLGA-Ery NPs were synthesized by the o/w emulsion diffusion method, having a particle size of 159 ± 23 nm and displayed 71.89 % of encapsulation efficiency. The PLGA-Ery NPs showed 1.5, 2.1 and 1.5-fold improved MIC and antibacterial efficacy against E. coli, S. aureus, and P. aeruginosa, respectively than the pure drug. As illustrated by scanning electron microscopy, PLGA-Ery NPs caused damage to the bacterial cell walls. Furthermore, a surface coating with PLGA-Ery NPs on a glass surface showed efficient inhibition (>90 %) of the biofilm formation by P. aeruginosa, as determined by fluorescence microscopy and MTT assay. This study demonstrates that PLGA-Ery NPs can increase the efficiency of erythromycin and can suppress the growth and biofilm formation of P. aeruginosa. Such polymeric nanoparticles drug nanoformulations have potential as an antimicrobial and as a surface coating for medical devices.

Investigating the antimicrobial and antibiofilm properties of marine halophilic Bacillus species against ESKAPE pathogens.

Antimicrobial resistance (AMR), known as the "silent pandemic," is exacerbated by pathogenic bacteria's ability to form biofilms. Marine compounds hold promise for novel antibacterial drug discovery. Two isolates from preliminary saltwater environment screening demonstrated antimicrobial activity and were subsequently identified as Bacillus subtilis MTUA2 and Bacillus velezensis MTUC2. Minimum inhibitory concentrations (MICs), minimum biofilm inhibition concentrations (MBICs) and minimum biofilm eradication concentrations (MBECs) required to prevent and/or disrupt bacterial growth and biofilm formation were established for MRSA, Staphylococcus aureus, Acinetobacter baumannii and Escherichia coli. The metabolic activity within biofilms was determined by the 2,3,5-triphenyltetrazolium chloride assay. Both Bacillus species exhibited unique antimicrobial effects, reducing MRSA and S. aureus planktonic cell growth by 50% and sessile cell growth for S. aureus and E. coli by 50% and 90%, respectively. No effect was observed against A. baumannii. Significant MBIC and MBEC values were achieved, with 99% inhibition and 90% reduction in MRSA and S. aureus biofilms. Additionally, 90% and 50% inhibition was observed in E. coli and A. baumannii biofilms, respectively, with a 50% reduction in E. coli biofilm. These findings suggest that the mode of action employed by B. subtilis MTUA2 and B. velezensis MTUC2 metabolites should be further characterized and could be beneficial if used independently or in combination with other treatments.