Biofilm Inhibition Research Articles

Manganese Complex-Gold Nanoparticle Hybrid for Biofilm Inhibition and Eradication.

Biofilms, which are resistant to conventional antimicrobial treatments, pose significant challenges in medical and industrial environments. This study introduces manganese complex-gold nanoparticles (Mn-DPA-AuNPs) as a hybrid strategy for biofilm inhibition and eradication. Upon exposure to green light, these nanoparticles significantly enhance the generation of reactive oxygen species (ROS), including hydrogen peroxide and superoxide. This activity substantially reduces the regrowth potential of the surviving bacteria within the biofilm, with marked efficacy noted in Pseudomonas aeruginosa PAO1. This study highlights the potential of integrating manganese complexes with gold nanoparticles to develop advanced antimicrobial agents against resistant biofilms, offering a promising approach to enhance microbial control in diverse settings.

The Potential of Thymus serpyllum Essential Oil as an Antibacterial Agent against Pseudomonas aeruginosa in the Preservation of Sous Vide Red Deer Meat.

Foodborne infections caused by microbes are a serious health risk. Regarding this, customer preferences for "ready-to-eat" or minimally processed (MP) deer meat are one of the main risk factors. Given the health dangers associated with food, essential oil (EO) is a practical substitute used to decrease pathogenic germs and extend the shelf-life of MP meals. Nonetheless, further data regarding EO use in MP meals are required. In order to evaluate new, safer alternatives to chemicals for disease control and food preservation, this research was carried out in the following areas to assess the antibacterial and antibiofilm characteristics of Thymus serpyllum (TSEO) essential oil, which is extracted from dried flowering stalks. Furthermore, this study applied an essential oil of wild thyme and inoculated the sous vide deer meat with Pseudomonas aeruginosa for seven days at 4 °C in an effort to prolong its shelf-life. Against P. aeruginosa, the essential oil exhibited potent antibacterial action. The findings of the minimal biofilm inhibition concentration (MBIC) crystal violet test demonstrated the substantial antibiofilm activity of the TSEO. The TSEO modified the protein profiles of bacteria on glass and plastic surfaces, according to data from MALDI-TOF MS analysis. Moreover, it was discovered that P. aeruginosa was positively affected by the antibacterial properties of TSEO. The anti-Pseudomonas activity of the TSEO was marginally higher in vacuum-packed sous vide red deer meat samples than in control samples. The most frequently isolated species from sous vide deer meat, if we do not consider the applied bacteria Pseudomonas aeruginosa, were P. fragi, P. lundensis, and P. taetrolens. These results highlight the antibacterial and antibiofilm qualities of TSEO, demonstrating its potential for food preservation and extending the shelf-life of deer meat.

Antibacterial and Anti-Inflammatory Activity of Branched Peptides Derived from Natural Host Defense Sequences.

Antibiotic resistance is a major global health threat, necessitating the development of new treatments and diverse molecules to combat severe infections and preserve the efficacy of existing drugs. Antimicrobial peptides (AMPs) offer a versatile arsenal against bacteria, and peptide structure branching can enhance their resistance to proteases and improve their overall efficacy. A small library of peptides derived from natural host defense peptides and synthesized in a tetrabranched form was selected against E. coli. Six selected branched peptides were further studied for antibacterial activity against a panel of strains, biofilm inhibition, protease resistance, and cytotoxicity. Their structure was predicted computationally and their mechanism of action was investigated by electron microscopy and by using fluorescent dyes. The peptide BAMP2 showed promise in a mouse skin infection model, indicating the potential for local infection treatment.

Antimicrobial Peptide Octoprohibitin-Encapsulated Chitosan Nanoparticles Enhanced Antibacterial Activity against Acinetobacter baumannii

Background: This study focused on evaluating the physiochemical characteristics and antibacterial activity of Octoprohibitin-encapsulated CNPs (Octoprohibitin-CNPs) against Acinetobacter baumannii. Methods: Octoprohibitin was encapsulated into CNPs via ionotropic gelation with carboxymethyl chitosan (CMC) and low molecular weight chitosan (CS). Octoprohibitin-CNPs were dispersed in phosphate-buffered saline and the release kinetic profile was determined. Then Octoprohibitin-CNPs were examined using field-emission transmission electron microscopy and physicochemical characterization was performed. Antibacterial activity of Octoprohibitin-CNPs against A. baumannii was evaluated. Biofilm inhibition and eradication assays were performed using the crystal violet (CV) staining-based method for biofilm quantification. Results: The average diameter, zeta potential, encapsulation efficiency, and loading capacity of Octoprohibitin-CNPs were 244.5 ± 21.97 nm, +48.57 ± 0.38 mV, and 85.7% and 34.2%, respectively. TEM analysis imaging revealed that Octoprohibitin-CNPs are irregularly shaped, with fewer aggregates than CNPs. Octoprohibitin-CNPs exhibited a biphasic release pattern, characterized by an initial rapid phase followed by a sustained release over time, extending up to 93.68 ± 6.48% total release until 96 h. In vitro, Octoprohibitin-CNPs showed lower cytotoxicity compared to Octoprohibitin alone. Time-kill kinetic and bacterial viability reduction assays showed Octoprohibitin-CNPs exhibited slightly higher antibacterial activity against A. baumannii than Octoprohibitin. Conclusions: Octoprohibitin-CNP-treated A. baumannii exhibited higher levels of morphological deviation, increased membrane permeability, and the production of reactive oxygen species, as well as antibiofilm activity with greater biofilm inhibition and eradication than Octoprohibitin. These findings show that Octoprohibitin-CNPs perform better against A. baumannii compared to Octoprohibitin alone.

Phytosynthesis of Silver Nanoparticles Using Mansoa alliacea (Lam.) A.H. Gentry (Bignoniaceae) Leaf Extract: Characterization and Their Biological Activities

Background. Mansoa alliacea is a native plant renowned for its medicinal properties in traditional healing in the Amazon Region. This plant is rich in polyphenols, flavonoids, anthocyanins, phenolic acids, tannins, ketones, triterpenes, as well as other bioactive compounds. Objectives. This study aims to develop an innovative, eco-friendly method for synthesizing silver nanoparticles using an aqueous extract of M. alliacea (Ma-AgNPs), enhancing the biological activities of AgNPs by leveraging the therapeutic potential of the plant’s bioactive compounds. Methods. Silver nanoparticles were synthesized using the aqueous extract of M. alliacea. The biological activities of Ma-AgNPs were assessed, including antibacterial, anti-inflammatory, antioxidant, antitumor, and anti-biofilm effects, along with evaluating their hemolytic activity. Results. Quantitative analysis revealed that Ma-AgNPs exhibit potent antibacterial activity against multidrug and non-multidrug-resistant bacteria, with MIC values ranging from 1.3 to 10.0 µg/mL. The Ma-AgNPs significantly reduced NO production by 86.9% at 4 µg/mL, indicating strong anti-inflammatory effects. They demonstrated robust antioxidant activity with an IC50 of 5.54 ± 1.48 µg/mL and minimal hemolytic activity, with no hemolysis observed up to 20 µg/mL and only 4.5% at 40 µg/mL. Their antitumor properties were notable, with IC50 values between 2.9 and 5.4 µg/mL across various cell lines, and they achieved over 50% biofilm inhibition at concentrations of 30–40 µg/mL. Conclusions. These findings underscore the potential of Ma-AgNPs for biomedical applications, particularly in developing new antimicrobial agents and bioactive coatings with reduced toxicity. This research highlights a sustainable approach that not only preserves but also amplifies the inherent biological activities of plant extracts, paving the way for innovative therapeutic solutions.

Exploring diflunisal as a synergistic agent against Staphylococcus aureus biofilm formation.

Staphylococcus aureus is a bacterial pathogen of considerable significance in public health, capable of inducing a diverse range of infectious diseases. One of the most notorious mechanisms used by S. aureus to survive and colonize the site of infection is its ability to form biofilms. Diflunisal, a non-steroidal anti-inflammatory drug (NSAID), is a known inhibitor of the Agr system in S. aureus, which is key in regulating biofilm formation. This study evaluated the effect of broad-spectrum antibiotics in combination with diflunisal on S. aureus biofilm density. Eight antibiotics were tested independently at different concentrations and in combination with diflunisal to assess their effect on S. aureus biofilm formation. When using the antibiotics alone and with diflunisal, a significant control effect on biofilm formation was observed (p < 0.05), irrespective of diflunisal presence, but did not achieve a complete biofilm growth inhibition. Over time, diflunisal influenced biofilm formation; however, such an effect was correlated with antibiotic concentration and exposure time. With amikacin treatments, biofilm density increased with extended exposure time. In the case of imipenem, doripenem, levofloxacin, and ciprofloxacin, lower doses and absence of diflunisal showed higher control over biofilm growth with longer exposure. However, in all cases, diflunisal did not significantly affect the treatment effect on biofilm formation. In the absence of antibiotics, diflunisal significantly reduced biofilm formation by 53.12% (p < 0.05). This study suggests that diflunisal could be a potential treatment to control S. aureus biofilms, but it does not enhance biofilm inhibition when combined with antibiotics.

Identification and preclinical evaluation of MMV676558 as a promising therapeutic candidate against Clostridioides difficile

Clostridioides difficile, a gram-positive, toxin-producing, spore-forming anaerobe, is a major cause of antibiotic-associated diarrhoea. The bacterium's intrinsic drug resistance limits current treatment options to fidaxomicin and vancomycin for initial episodes, with anti-toxin B monoclonal antibody or faecal microbiota transplantation recommended for complicated or recurrent cases. This underscores the urgent need for novel therapeutics. In this study, we screened the MMV Pathogen Box at a 10 µM concentration against C. difficile R20291. Primary hits were evaluated for minimum inhibitory concentrations (MIC), killing kinetics, and biofilm inhibition. Bacterial cytological profiling (BCP) and transmission electron microscopy (TEM) were employed to study the mode of action. MMV676558 was further tested in a mouse model to assess survival, histopathology, and gut microbiota effects. We identified nineteen hits that inhibited over 50 % of C. difficile growth. MIC assays revealed three hits with MICs below 16 µg/mL: MMV676558, MMV688755, and MMV690027. These hits were effective against various C. difficile ribotypes. Killing kinetics were comparable or superior to vancomycin and fidaxomicin, and biofilm assays showed inhibitory effects. BCP and TEM analyses suggested membrane function disruption as the mode of action. Furthermore, MMV676558 demonstrated a protective effect in mice, with favourable histopathology and gut microbiota profiles. Given the urgent threat posed by C. difficile antibiotic resistance, discovering new treatments is a top priority. Our study identified three promising hits from the MMV Pathogen Box, with MMV676558 showing significant in vivo potential for further evaluation.

Green-synthesized silver nanoparticles from Zataria multiflora as a promising strategy to target quorum sensing and biofilms in Pseudomonasaeruginosa

The global challenge to human health is significantly heightened by the resistance of harmful bacteria to antimicrobial treatments. Given the limited advancement in developing new antimicrobial medications, exploring innovative strategies is imperative to tackle the challenge of resistance to multiple drugs. Furthermore, there is a growing emphasis on the environmentally friendly synthesis of nanoparticles with potent medicinal attributes, specifically those targeting virulence, to combat the rise of multidrug resistance. Focusing on the inhibition of virulence factors and biofilms influenced by quorum sensing has become a promising and novel strategy in the development of anti-infective drugs. An aqueous extract of Zataria multiflora leaves was used to create green-synthesized silver nanoparticles, or AgNPs. X-ray diffraction (XRD), transmission electron microscopy (TEM), scanning electron microscopy (SEM), Fourier-transform infrared spectroscopy (FTIR), and UV–visible absorption spectroscopy were used to characterize the AgNPs. The impact of AgNPs on the virulence factors and biofilms of Pseudomonas aeruginosa PAO1, mediated by quorum sensing, was assessed at concentrations below the minimum inhibitory concentration (sub-MIC). Sub-MIC concentrations of Green-synthesized AgNPs inhibited various P. aeruginosa virulence factors, including bacterial motility (89 % inhibition), pyocyanin production (81.48 % inhibition), pyoverdin production (55.80 % inhibition), elastase activity (87.43 % inhibition), exoprotease activity (75.60 % inhibition), and rhamnolipid production (71.28 % inhibition). Additionally, these AgNPs demonstrated 80 % inhibition of P. aeruginosa biofilms. The in vitro efficacy of green-synthesized AgNPs against P. aeruginosa can be utilized for the creation of alternative therapeutic agents for managing bacterial infections, particularly for topical application in cases such as wound infections. Additionally, they can be used for surface coating to inhibit the attachment of bacteria to medical devices.

Potential of silver nanoparticles synthesized from Justicia adhatoda metabolites for inhibiting biofilm on urinary catheters

In the present study, we investigated the anti-biofilm effect of urinary catheters fabricated with biogenic nanoparticles synthesized from metabolites of Justicia adhatoda under in vitro conditions against human pathogenic bacteria. Silver nanoparticles were synthesized in the reaction mixture composed of 2 % w/v of 0.1 M of precursor (silver nitrate) and 0.2 g of the metabolites obtained from ethanolic extract of Justicia adhatoda. Characterization of the nanoparticles was done by UV visible spectroscopy, fourier infrared spectroscopy (FTIR), scanning electron microscopy (SEM) and X ray diffraction (XRD) to confirm the structural and functional properties. Primary conformation of nanoparticles synthesis by UV visible spectroscopy revealed the notable absorption spectra at 425 nm with a wavelength shift around 450 nm, likely due to surface plasmon resonance excitation. SEM analysis showed spherical, monodisperse, nano scale particles with a size range of 50–60 nm. Crystaline phase of the synthesized nanoparticles was confirmed by x ray diffraction studies which showed the distinct peaks at (2θ) 27.90, 32.20, 46.30, 54.40, and 67.40, corresponding to (111), (200), (220), (222), and (311) planes of nano scale silver. The biocompatibility of these nanoparticles was assessed through zebrafish embryonic toxicity study which showed more than 90 % of embryos were alive and healthy. No marked changes on the blood cells also confirmed best hemocompatibility of the nanoparticles. Synthesized nanoparticles thus obtained were fabricated on the urinary catheter and the fabrication was confirmed by FTIR and SEM analysis. Notable changes in the absorption peaks, uniform coating and embedding of silver nanoparticles studied by FTIR and SEM analysis confirmed the fabrication of silver nanoparticles. The coated catheters demonstrated significant antibacterial activity against pathogenic bacterial strains, including E. coli ATCC 25922 and Pseudomonas aeruginosa ATCC 27853. Anti-biofilm studies, conducted using a modified microtiter plate crystal violet assay, revealed effective inhibition of both bacterial adhesion and biofilm development. 85 % of biofilm inhibition was recorded against both the tested strains. The coating method presented in this study shows promise for enhancing infection resistance in commonly used medical devices like urinary catheters, thus addressing device-associated infections.

Synergetic inhibitory effect of isopropyl methylphenol-based agents on biofilm formation by Streptococcus mutans.

Dental caries and periodontitis are the most common oral diseases in humans and the main causes of tooth loss. Streptococcus mutans is primarily responsible for dental caries and dental plaque, which are triggered by biofilm formation on the tooth surface. In this study, biofilm inhibition by 4-isopropyl-3-methylphenol (IPMP)-based agents, consisting of IPMP and polyoxyethylene-hydrogenated castor oil (POEHCO), was investigated in vitro. Notably, the use of POEHCO in addition to IPMP inhibited S. mutans biofilms more drastically than IPMP alone. Moreover, the effects of IPMP on the expression of biofilm-related genes (gtfB, gtfC, and gtfD) were examined using quantitative real-time PCR. IPMP at sub-minimum inhibitory concentrations significantly downregulated the expression of these genes. These results suggested that the inhibitory effects on biofilm formation were synergistically enhanced by the surfactant and antibiofilm activities of IPMP. Therefore, IPMP-based agents as dentifrices may be useful to prevent oral diseases originating from biofilms.

Chirality differences of three borneols contribute to controlling Escherichia coli biofilm formation by chemotaxis mediating movement

SummaryEscherichia coli (E. coli) biofilm is the major cause of its high lethality in foodborne illness. Controlling E. coli biofilm formation is a critical way to reduce foodborne diseases. Here, chiral borneol isomers (L‐borneol, D‐borneol, and isoborneol) were identified as E. coli chemorepellents, which reduced reversible adhesion at the initial stage of biofilm formation. Borneols directly inhibited biofilm production and their inhibitory capacities were associated with the chiral stereochemical structures (L‐borneol &gt; D‐borneol &gt; isoborneol). A transcriptomic study revealed the inhibitory mechanism of borneols was attributed to the bacterial motility intention and motility energy system. The most effective isomer, L‐borneol, reduced bacterial energy metabolism and increased E. coli chemotaxis motility intention through the up‐regulation of genes associated with flagellar assembly and bacterial chemotaxis pathways. This work demonstrates that borneols are biofilm inhibitors and supports the potential of borneols as a stereochemical antimicrobial strategy to reduce foodborne pathogenic bacterial contamination.

Cellulose acetate-based electrospun nanocomposites improved by mussel-inspired polydopamine coatings and copper iodide-decorated graphene oxide: A self-disinfecting nanofibrous membrane with potential biomedical applications

Personal protective equipment and fabrics have proven their efficient use in the SARS-CoV-2 pandemic, but because germs and viruses may survive on them, they can also operate as a source of disease transmission. Self-disinfecting textiles with the potential to kill different pathogens can be used as a substitute and prevent infections. In this research, copper iodide (CuI) decorated graphene oxide nanocomposite was synthesized through a green water-based procedure, and a novel nanofibrous membrane with improved antibacterial and antiviral characteristics was prepared by a mussel-inspired method, in which polydopamine (DOPA) and CuI-decorated graphene oxide (GO-CuI) nanocomposite were coated on cellulose acetate electrospun nanofibers (CA-DOPA-GOCuI). CA-DOPA-GOCuI NFM displayed extremely good antibacterial/antiviral properties with biofilm inhibition of 50 %, and very good cell viability and proliferation even though the virus was introduced to the cell culture. Furthermore, because it accelerated the healing process, this NFM showed great promise for use as a wound dressing. This may be because it inhibits infection and has an extremely high ROS scavenging capability of 80 %. Additionally, it was shown that CA-DOPA-GOCuI NFM was non-toxic to cells and blood compatible, with cell viability that was even higher than that of the control group of untreated cells. One possible explanation for this could be the inclusion of GO, a highly biocompatible member of the graphene family, which, when combined with CuI, reduced its toxicity to normal cells. Thus, the safety of CA-DOPA-GOCuI to normal cells, favorable cell proliferation and viability, and its high toxicity to bacteria and viruses indicate its great potential to be used in a wide range of biomedical applications, specifically, self-disinfecting fabrics and wound dressings.

Bacteriophage P2-71: a promising therapeutic against multidrug-resistant Proteus mirabilis in urinary tract infections.

Proteus mirabilis is a Gram-negative, rod-shaped bacterium widely found in natural environments. It is known for causing a range of severe illnesses in mammals, particularly urinary tract infections (UTIs). This study evaluates the therapeutic efficacy of phage P2-71 against Proteus mirabilis in vivo and in vitro environments. The in vitro therapeutic potential of bacteriophage P2-71 was assessed through the ability of phage to kill Proteus mirabilis by using a plate counting assay, and biofilm inhibition and biofilm lysis assays using a microtitre plate method. Additionally, an in vivo UTI model in C57BL/6Jmice was developed via urethral inoculation of the bacterium. Phage therapy was administered through urethral injection over a period of 5 days. Therapeutic outcomes were measured by analyzing bacterial load, phage titer, inflammatory markers, and histopathological changes in the urine, urogenital tissues, and spleen. In vitro, bacteriophage P2-71 achieved significant reductions in P. mirabilis concentrations, with log reductions of 1.537 and 0.7009 CFU/mL in laboratory and urine environments, respectively (p < 0.001). The phage also decreased biofilm formation by 34-49% and lysed 15-25% of mature biofilms at various multiplicities of infection (MOIs) (p < 0.001). In vivo, phage treatment significantly lowered bacterial concentrations in the urine on Days 1 and 3 (p < 0.0001), achieving a maximum reduction of 4.602 log₁₀ CFU/mL; however, its effectiveness diminished by Day 5 (p > 0.05). Concurrently, phage titers decreased over time. Importantly, phage treatment notably reduced bacterial load in the bladder, kidneys, and spleen (p < 0.001). Inflammatory markers such as IL-6, IL-1β, and TNF-α were significantly lower in the treatment group, especially in the bladder (p < 0.0001), indicating an effective reduction in inflammation. Histopathological analysis showed significant mitigation of tissue damage. The results demonstrated that bacteriophage P2-71 is a promising alternative therapy for UTIs caused by MDR Proteus mirabilis. This bacteriophage therapy offers a viable strategy for managing infections where traditional antimicrobials fail, highlighting its potential in clinical applications.

Hybrids of 3-Hydroxypyridin-4(1H)-ones and Long-Chain 4-Aminoquinolines as Potent Biofilm Inhibitors of Pseudomonas aeruginosa Potentiate Tobramycin and Polymyxin B Activity.

The biofilm formation of Pseudomonas aeruginosa involves multiple complex regulatory pathways; thus, blocking a single pathway is unlikely to achieve the desired antibiofilm efficacy. Herein, a series of hybrids of 3-hydroxypyridin-4(1H)-ones and long-chain 4-aminoquinolines were synthesized as biofilm inhibitors against P. aeruginosa based on a multipathway antibiofilm strategy. Comprehensive structure-activity relationship studies identified compound 30b as the most valuable antagonist, which significantly inhibited P. aeruginosa biofilm formation (IC50 = 5.8 μM) and various virulence phenotypes. Mechanistic studies revealed that 30b not only targets the three quorum sensing systems but also strongly induces iron deficiency signals in P. aeruginosa. Furthermore, 30b demonstrated a favorable in vitro and in vivo safety profile. Moreover, 30b specifically enhanced the antibacterial activity of tobramycin and polymyxin B in in vitro and in vivo combination therapy. Overall, these results highlight the potential of 30b as a novel anti-infective candidate for treating P. aeruginosa infections.

In Vitro Probiotic Characterization of Yeasts with their Postbiotics' Antioxidant Activity and Biofilm Inhibition Capacity.

This study evaluated the in vitro probiotic potential and postbiotic properties of yeast strains isolated from traditional fermented foods, emphasizing antioxidant activity (AOA) and biofilm inhibition capacity (BIC). The yeasts were molecularly confirmed using start codon targeted polymorphisms as Kluyveromyces lactis (n = 17), Saccharomyces cerevisiae (n = 9), Pichia kudriavzevii (n = 6), P. fermentans (n = 4), Wickerhamomyces anomalus (n = 2), and Torulaspora delbrueckii (n = 1). The probiotic assessment of live cells included viability in simulated gastric and pancreatic juices, autoaggregation, hydrophobicity, and AOA, using S. boulardii MYA-796 as reference. Additionally, cell-free supernatants (CFS) were tested for AOA and BIC against Cronobacter sakazakii, Listeria monocytogenes, Pseudomonas aeruginosa, and Staphylococcus aureus. Several strains exhibited significantly higher in vitro probiotic characteristics compared to S. boulardii MYA-796 (P < 0.05), particularly in gastric and pancreatic survival, hydrophobicity, and AOA. Notably, CFS exhibited greater AOA than live cells and strong BIC, especially against L. monocytogenes and S. aureus. Multivariate analysis identified K. lactis TC11, S. cerevisiae M33T1-2, P. kudriavzevii S96, W. anomalus OB7Y1, and T. delbrueckii KY31 as having superior probiotic properties, attributed to enhanced gastric survival, autoaggregation, and AOA. CFS of S. cerevisiae M33T1-2 and T. delbrueckii KY31 demonstrated significant BIC, with over 60% inhibition across all tested pathogens.

Enhanced Antibacterial Activity of Carbon Dots: A Hybrid Approach with Levofloxacin, Curcumin, and Tea Polyphenols

Bacterial infections and their increasing resistance to antibiotics pose a significant challenge in medical treatment. This study presents the synthesis and characterization of novel carbon dots (CDs) using levofloxacin (Lf), curcumin (Cur), and tea polyphenols (TP) through a facile hydrothermal method. The synthesized curcumin-tea polyphenol@carbon dots (Cur-TP@CDs) and levofloxacin-tea polyphenol@carbon dots (Lf-TP@CDs) were characterized using transmission electron microscopy (TEM), Fourier transform infrared spectroscopy (FT-IR), X-ray diffraction (XRD), X-ray photoelectron spectroscopy (XPS), and Raman spectroscopy, confirming their unique structural and chemical properties. Cur-TP@CDs exhibited an average particle size of 1.32 nanometers (nm), while Lf-TP@CDs averaged 1.58 nm. Both types demonstrated significant antibacterial activity, with Lf-TP@CDs showing superior effectiveness against Staphylococcus aureus (S. aureus) and Escherichia coli (E. coli) in broth dilution and disc diffusion assays. Biofilm inhibition assays revealed a significant reduction in biofilm formation at higher concentrations. The ultraviolet-visible (UV-vis) and photoluminescence (PL) spectral analyses indicated efficient photon emission, and electron paramagnetic resonance (EPR) analysis showed increased singlet oxygen generation, enhancing bactericidal effects. Live and dead bacterial staining followed by scanning electron microscopy (SEM) analysis confirmed dose-dependent bacterial cell damage and morphological deformities. These findings suggest that Cur-TP@CDs and Lf-TP@CDs are promising antibacterial agents, potentially offering a novel approach to combat antibiotic-resistant bacterial infections.

Actinomycin D reduces virulence factors and biofilms against Aeromonas hydrophila.

Aeromonas hydrophila, a Gram-negative bacterium, is ubiquitously found in many aquatic habitats, causing septicemia in humans and fishes. Attributed to abuse or misuse of conventional antimicrobial drug usage, antimicrobial resistance is at an alarming rise. There is an available alternative strategy to bacterial resistance to antimicrobials, which is inhibition of virulence and pathogenicity employing quorum sensing inhibitors (QSIs). Hence, actinomycin D's effectiveness against A. hydrophila SHAe 115 as a QSI was investigated in decreasing virulence factors and preventing biofilm formation. Actinomycin D, belongs to the QSI combating Pseudomonas aeruginosa PAO1 originally isolated from an entophytic actinomycete (Streptomyces cyaneochromogenes RC1) in Areca catechu L. In the present work, further investigations were carried out to assess the effect of actinomycin D at subminimal inhibitory concentrations (sub-MICs), QS-regulated virulence factors, and biofilm inhibition strategies. Intrinsic properties encompassing inhibition of the production of protease and hemolysin and subsequent activities on biofilm formation and eradication of mature biofilm were established along with weakened swimming and swarming motilities in A. hydrophila SHAe 115. In the Tenebrio molitor survival assay, actinomycin D effectively reduced the virulence and pathogenicity of A. hydrophila, resulting in elimination of mortality. However, the hydrolysate of actinomycin D, 2-hydroxy-4,6-dimethyl-3-oxo-3H-phenoxazine-1,9-dicarboxylic acid (HDPD), had lost the QSI activity in A. hydrophila. Actinomycin D was proved as a viable QSI in lessening A. hydrophila's the virulence and pathogenicity, as evident from our research findings.

Assessment of the potential susceptibility of planktonic cells and bacterial biofilms by diosgenin

Bacterial biofilms are formed by a complex community of microorganisms, held together by a matrix that provides structural support and stability to microbial communities, making infections associated with biofilms more difficult to treat. In the face of growing research in the health area in search of antibiotics with action against biofilms, natural products present an effective alternative strategy. In this context, the aim of this study was to evaluate bacterial biofilm formation, assess the susceptibility of planktonic cells to diosgenin, and examine its efficacy on inhibiting and treating biofilms. The broth microdilution method was used to determine the Minimum Inhibitory Concentration (MIC). Six bacterial strains were tested: Staphylococcus aureus ATCC 25923, Pseudomonas aeruginosa ATCC 9027, Pseudomonas aeruginosa 24, Enterococcus faecalis INCQS 0017, Enterococcus faecalis INCQS 00018, and Enterococcus faecalis INCQS 654. The crystal violet method was employed to evaluate the potential of diosgenin in biofilm inhibition and eradication. Chlorhexidine gluconate served as a reference standard for the antibiofilm assays, along with the antibiotics norfloxacin, ampicillin, and gentamicin. MIC determination revealed that diosgenin exhibited intrinsic antimicrobial activity, with an MIC of 406 μg/mL against S. aureus ATCC 25923. The antibiofilm activity showed significant inhibition for E. faecalis 17 and moderate inhibition against E. faecalis 654 and S. aureus 25923. Furthermore, diosgenin effectively eradicated preformed biofilms of S. aureus 25923 and P. aeruginosa 9027, indicating its ability to interfere with biofilm biomass. These results suggest that diosgenin not only inhibits bacterial biofilm formation but also eradicates established biofilms, highlighting it as a promising candidate for the development of products aimed at combating bacterial biofilms.

Exploring Biofilm-Related Traits and Bile Salt Efficacy as Anti-Biofilm Agents in MDR Acinetobacter baumannii.

Acinetobacter baumannii has been designated as a critical priority pathogen by the World Health Organization for the development of novel antimicrobial agents. This study aimed to investigate both the phenotypic and genotypic traits of multidrug-resistant (MDR) A. baumannii strains, along with the effects of natural bile salts on biofilm formation. The research analyzed phenotypic traits, including autoaggregation, hydrophobicity, twitching motility, lectin production, and biofilm formation, as well as genotypic traits such as the presence of bap and blaPER-1 genes in twenty wound and eight environmental MDR A. baumannii isolates. While all strains were identified as good biofilm producers, no statistically significant correlation was detected between the examined traits and biofilm formation. However, differences in biofilm production were observed between environmental and wound isolates. The natural bile salts Na-cholate, Na-deoxycholate, and Na-chenodeoxycholate demonstrated effective anti-A. baumannii activity (MIC = 0.25-10 mg mL-1), with significant anti-biofilm effects. Na-deoxycholate and Na-chenodeoxycholate inhibited 94-100% of biofilm formation at super-MIC concentrations (8-32 mg mL-1). This study underscores the urgent need for innovative strategies to combat antibiotic resistance and biofilm formation in A. baumannii, highlighting the potential of natural bile salts as promising biofilm inhibitors and encouraging further research into their modification and combination with other antimicrobials.

Mining for antifungal agents to inhibit biofilm formation of Candida albicans: A study on green synthesis, antibiofilm, cytotoxicity, and in silico ADME analysis of 2-amino-4H-pyran-3-carbonitrile derivatives

Candida albicans (C. albicans) biofilm infections are quite difficult to manage due to their resistance against conventional antifungal drugs. To address this issue, there is a desperate need for new therapeutic drugs. In the present study, a green and efficient protocol has been developed for the synthesis of 2-amino-4H-pyran-3-carbonitrile scaffolds 4a-i, 6a-j, and 8a-g by Knoevenagel-Michael-cyclocondensation reaction between aldehydes, malononitrile, and diverse enolizable C-H activated acidic compounds using guanidinium carbonate as a catalyst either under grinding conditions or by stirring at room temperature. This protocol is operationally simple, rapid, inexpensive, has easy workup and column-free purification. A further investigation of the synthesized compounds was conducted to examine their antifungal potential and their ability to inhibit the growth and development of biofilm-forming yeasts like fungus C. albicans. According to our findings, 4b, 4d, 4e, 6e, 6f, 6g, 6i, 8c, 8d, and 8g were found to be active and potential inhibitors for biofilm infection causing C. albicans. The inhibition of biofilm by active compounds were observed using field emission scanning electron microscopy (FESEM). Biofilm inhibiting compounds were also tested for in vitro toxicity by using 3T3-L1 cell line, and 4b, 6e, 6f, 6g, 6i, 8c, and 8d were found to be biocompatible. Furthermore, the in silico ADME descriptors revealed drug-like properties with no violation of Lipinski's rule of five. Hence, the result suggested that synthesized derivatives could serve as a useful aid in the development of novel antifungal compounds for the treatment of fungal infections and virulence in C. albicans.