Antibiofilm Agents Research Articles

Chemical composition, cytotoxicity, antimicrobial, antibiofilm, and anti-quorum sensing potential of Mentha Piperita essential oil against the oral pathogen Streptococcus mutans

ABSTRACT Dental caries is a highly prevalent oral disease affecting billions of individuals globally. The disease occurs chemically as a result of breakdown of the tooth surface attributed to metabolic activity in colonizing biofilm. Biofilms, composed of exopolysaccharides and proteins, protect bacteria like Streptococcus mutans, which is notable for its role in tooth decay due to its acid-producing abilities. While various antimicrobial agents may prevent biofilm formation, these drugs often produce side effects including enamel erosion and taste disturbances. This study aimed to examine utilization of the Mentha piperita essential oil as a potential antibiofilm activity agent against S. mutans. M. piperita oil significantly (1) reduced bacterial biofilm, (2) exhibited a synergistic effect when combined with chlorhexidine, and (3) did not induce cell toxicity. Chemical analysis identified the essential oil with 99.99% certainty, revealing menthol and menthone as the primary components, constituting approximately 42% and 26%, respectively. Further, M. piperita oil eradicated preformed biofilms and inhibited biofilm formation at sub-inhibitory concentrations. M. piperita oil also interfered with bacterial quorum sensing communication and did not produce any apparent cell toxicity in immortalized human keratinocytes (HaCaT). M. piperita represented an alternative substance for combating S. mutans and biofilm formation and a potential combination option with chlorhexidine to minimize side effects. An in-situ performance assessment requires further studies.

Mentha longifolia assisted nanostructures: An approach to obliterate microbial biofilms.

Microbes maneuver strategies to become incessant and biofilms perfectly play a role in scaling up virulence to cause long-lasting infections. The present study was designed to assess the use of an eco-friendly formulation of functionalized silver nanoparticles generated from Mentha longifolia leaf extract (MℓE) for the treatment of biofilm-producing microbes. Nanoparticles synthesized using MℓE as a reducing agent were optimized at different strengths of AgNO3 (1 mM, 2 mM, 3 mM, and 4 mM). Synthesis of M. longifolia silver nanoparticles (MℓAgNPs) was observed spectrophotometrically (450 nm) showing that MℓAgNPs (4 mM) had the highest absorbance. Various techniques e.g., Fourier transforms Infrared spectroscopy (FTIR), Dynamic light scattering (DLS), zeta potential (ZP), X-ray Diffraction (XRD), scanning electron microscope (SEM), and transmission electron microscope (TEM) were used to characterize MℓAgNPs. In the present study, the Kirby-Bauer method revealed 4mM was the most detrimental conc. of MℓAgNPs with MIC and MBC values of 0.62 μg/mL and 1.25 μg/mL, 0.03 μg/mL and 0.078 μg/mL, and 0.07 μg/mL and 0.15 μg/mL against previously isolated and identified clinical strains of Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumoniae, and Staphylococcus aureus, respectively. Moreover, the MℓAgNP antibiofilm activity was examined via tissue culture plate (TCP) assay that revealed biofilm inhibition of up to 87.09%, 85.6%, 83.11%, and 75.09% against E. coli, P. aeruginosa, K. pneumonia, and S. aureus, respectively. Herbal synthesized silver nanoparticles (MℓAgNPs) tend to have excellent antibacterial and antibiofilm properties and are promising for other biomedical applications involving the extrication of irksome biofilms. For our best knowledge, it is the first study on the use of the green-synthesized silver nanoparticle MℓAgNP as an antibiofilm agent, suggesting that this material has antibiotic, therapeutic, and industrial applications.

D-Methionine-induced DNases disperse established Burkholderia pseudomallei biofilms and promotes ceftazidime susceptibility

Burkholderia pseudomallei biofilm is correlated with pathogenesis, antibiotic resistance, and relapsing cases of melioidosis, leading to challenges in clinical management. There is increasing interest in employing biofilm dispersal agents as adjunctive treatments for biofilm-associated infections. Methionine (Met) has shown promise as an anti-biofilm agent by inducing bacterial DNase production, resulting in the degradation of extracellular DNA (eDNA) and dispersion of bacterial biofilm. In this study, we investigated the impact of 0.05–50 μM D-Met and L-Met on the 24-h established biofilm of a clinical isolate, B. pseudomallei H777. Our findings revealed the ability of D-Met and L-Met to disperse the established biofilm in a non-dose-dependent manner accompanied by eDNA depletion. Real-time PCR analysis further identified an up-regulation of bacterial nuclease genes, including recJ, eddB, nth, xth, and recD, in the presence of 0.05 μM D-Met. Similarly, recJ and eddB in B. pseudomallei were up-regulated in response to the presence of 0.05 μM L-Met. Notably, D-Met enhanced the susceptibility of B. pseudomallei H777 biofilm cells to ceftazidime. Our findings indicate a correlation between methionine supplementation and the up-regulation of nuclease genes, leading to eDNA depletion and the dispersal of preformed B. pseudomallei H777 biofilm. This enhances the susceptibility of biofilm cells to ceftazidime, showing promise in combating biofilm-associated B. pseudomallei infections.

FTY720 Reduces the Biomass of Biofilms in Pseudomonas aeruginosa in a Dose-Dependent Manner.

Pseudomonas aeruginosa, a nosocomial pathogen, has strong biofilm capabilities, representing the main source of infection in the human body. Repurposing existing drugs has been explored as an alternative strategy to combat emerging antibiotic-resistant pathogens. Fingolimod hydrochloride (FTY720), an immunomodulatory drug for multiple sclerosis, has shown promising antimicrobial effects against some ESKAPE pathogens. Therefore, the effects of FTY720 on the biofilm capabilities of Pseudomonas aeruginosa were investigated in this study. It was determined that FTY720 inhibited the growth of P. aeruginosa PAO1 at 100 µM. The significant reduction in PAO1 cell viability was observed to be dose-dependent. Additional cytotoxicity analysis on human cell lines showed that FTY720 significantly reduced viabilities at sub-inhibitory concentrations of 25-50 µM. Microtiter assays and confocal analysis confirmed reductions in biofilm mass and thickness and the cell survivability ratio in the presence of FTY720. Similarly, virulence production and biofilm-related gene expression (rhlA, rhlB, pilA, pilI, fliC, fliD and algR) were determined. The results demonstrate that pigment production was affected and quantitative real-time PCR analysis showed a variable degree of reduced gene expression in response to FTY720 at 12.5-50 µM. These findings suggest that FTY720 could be repurposed as an alternative antibiofilm agent against Pseudomonas aeruginosa.

Biogenic amine tryptamine in human vaginal probiotic isolates mediates matrix inhibition and thwarts uropathogenic E. coli biofilm

Probiotics offer a promising prophylactic approach against various pathogens and represent an alternative strategy to combat biofilm-related infections. In this study, we isolated vaginal commensal microbiota from 54 healthy Indian women to investigate their probiotic traits. We primarily explored the ability of cell-free supernatant (CFS) from Lactobacilli to prevent Uropathogenic Escherichia coli (UPEC) colonization and biofilm formation. Our findings revealed that CFS effectively reduced UPEC’s swimming and swarming motility, decreased cell surface hydrophobicity, and hindered matrix production by downregulating specific genes (fimA, fimH, papG, and csgA). Subsequent GC–MS analysis identified Tryptamine, a monoamine compound, as the potent bioactive substance from Lactobacilli CFS, inhibiting UPEC biofilms with an MBIC of 4 µg/ml and an MBEC of 8 µg/ml. Tryptamine induced significant changes in E. coli colony biofilm morphology, transitioning from the Red, Dry, and Rough (RDAR) to the Smooth and White phenotype, indicating reduced extracellular matrix production. Biofilm time-kill assays demonstrated a four-log reduction in UPEC viability when treated with Tryptamine, highlighting its potent antibacterial properties, comparable to CFS treatment. Biofilm ROS assays indicated a significant elevation in ROS generation within UPEC biofilms, suggesting a potential antibacterial mechanism. Gene expression studies with Tryptamine-treated samples showed a reduction in expression of curli gene (csgA), consistent with CFS treatment. This study underscores the potential of Tryptamine from probiotic Lactobacilli CFS as a promising antibiofilm agent against UPEC biofilms.

Anti-biofilm effect of enzymatic hydrolysates of ovomucin in Listeria monocytogenes and Staphylococcus aureus

Despite modern advances in food hygiene, food poisoning due to microbial contamination remains a global problem, and poses a great threat to human health. Especially, Listeria monocytogenes and Staphylococcus aureus are gram-positive bacteria found on food-contact surfaces with biofilms. These foodborne pathogens cause a considerable number of food poisoning and infections annually. Ovomucin (OM) is a water-insoluble gel-type glycoprotein in egg whites. Enzymatic hydrolysis can be used to improve the bioactive properties of OM. This study aimed to investigate whether ovomucin hydrolysates (OMHs) produced using five commercial enzymes (Alcalase®, Bromelain, α-Chymotrypsin, Papain, and Pancreatin) can inhibit the biofilm formation of L. monocytogenes ATCC 15313, L. monocytogenes H7962, S. aureus KCCM 11593, and S. aureus 7. Particularly, OMH prepared with papain (OMPP; 500 μg/mL) significantly inhibited biofilm formation in L. monocytogenes ATCC 15313, L. monocytogenes H7962, S. aureus KCCM 11593, and S. aureus 7 by 85.56 %, 80.28 %, 91.70 %, and 79.00 %, respectively. In addition, OMPP reduced the metabolic activity, exopolysaccharide production (EPS), adhesion ability, and gene expression associated with the biofilm formation of these bacterial strains. These results suggest that OMH, especially OMPP, exerts anti-biofilm effects against L. monocytogenes and S. aureus. Therefore, OMPP can be used as a natural anti-biofilm agent to control food poisoning in the food industry.

Comparison of the Proteome of Staphylococcus aureus Planktonic Culture and 3-Day Biofilm Reveals Potential Role of Key Proteins in Biofilm

Staphylococcus aureus and coagulase-negative staphylococci account for about 80% of infections associated with medical devices and are associated with increased virulence due to their ability to form biofilm. In this study, we aimed to construct a comprehensive reference map followed by significant pathway analysis in the proteome of S. aureus biofilm grown for 3 days compared with 24 h of planktonic culture using a high-resolution Tandem Mass Tag (TMT)-based MS. We identified proteins associated with secondary metabolites, ABC transporters, biosynthesis of amino acids, and response to stress, and amino sugar and nucleotide sugar metabolism were significantly upregulated in 3-day biofilm. In contrast, proteins associated with virulence factors, microbial metabolism in diverse environments, secondary metabolites, translation, and energy metabolism were significantly downregulated. GO functional annotation indicated that more proteins are involved in metabolic processes, catalytic activity, and binding in biofilm, respectively. Among the significantly dysregulated proteins, hyaluronidase (hysA) in conjunction with chitinase may play a significant role in the elimination and/or prevention of biofilm development. This study advances the understanding of the S. aureus subproteome, identifying potential pathways significant to biofilm biology. The insights gained may aid in developing new therapeutic strategies, including antibiofilm agents, for treating biofilm-related infections associated with implantable medical devices.

The Promising Effect of Ascorbic Acid and Paracetamol as Anti-Biofilm and Anti-Virulence Agents against Resistant Escherichia coli.

Escherichia coli is a major cause of serious infections, with antibiotic resistance rendering many treatments ineffective. Hence, novel strategies to combat this pathogen are needed. Anti-virulence therapy is a promising new approach for the subsequent era. Recent research has examined the impact of sub-inhibitory doses of ascorbic acid and paracetamol on Escherichia coli virulence factors. This study evaluated biofilm formation, protease production, motility behavior, serum resistance, expression of virulence-regulating genes (using RT-PCR), and survival rates in a mouse model. Ascorbic acid significantly reduced biofilm formation, protease production, motility, and serum resistance from 100% in untreated isolates to 22-89%, 10-89%, 2-57%, and 31-35% in treated isolates, respectively. Paracetamol also reduced these factors from 100% in untreated isolates to 16-76%, 1-43%, 16-38%, and 31-35%, respectively. Both drugs significantly down-regulated virulence-regulating genes papC, fimH, ompT_m, stcE, fliC, and kpsMTII. Mice treated with these drugs had a 100% survival rate compared with 60% in the positive control group control inoculated with untreated bacteria. This study highlights the potential of ascorbic acid and paracetamol as anti-virulence agents, suggesting their use as adjunct therapies alongside conventional antimicrobials or as alternative treatments for resistant Escherichia coli infections.

Characterization of Lemongrass Extract (Cymbopagon citratus) Nanoemulsion and Its Application as an Antibiofilm Agent in Acrylic Resin.

An antimicrobial agent is needed for denture cleaning, such as lemongrass (LG), which has a bioactive antimicrobial component. This research analyzed LG extract nanoparticles with a particle size analyzer, ZPA, and biofilm formation inhibition on resin acrylic surfaces. We found that there is high stability in nanoparticle size, while other concentrations, including chlorhexidine as a positive control, did not show any statistical differences. Lemongrass oil nanoemulsion has proved to be an antibiofilm and effective as a denture cleaning agent because of its ability to inhibit Streptococcus mutans and Candida albicans growth.

Mycobacterium tuberculosis Biofilms: Immune Responses, Role in TB Pathology, and Potential Treatment.

Tuberculosis (TB) is a major public health problem worldwide, and the burden of drug-resistant TB is rapidly increasing. Although there are literatures about the Mtb biofilms, their impact on immune responses has not yet been summarized. This review article provides recent knowledge on Mycobacterium tuberculosis (Mtb) biofilm-immunity interactions, their importance in pulmonary TB pathology, and immune-based therapy targeting Mtb biofilms. Pellicle/biofilm formation in Mtb contributes to drug resistance, persistence, chronicity, surface attachment, transfer of resistance genes, and modulation of the immune response, including reduced complement activation, changes in the expression of antigenic proteins, enhanced activation of T-lymphocytes, elevated local IFNγ+ T cells, and strong antibody production. The combination of anti-TB drugs and anti-biofilm agents has recently become an effective strategy to improve TB treatment. Additionally, immune-targeted therapy and biofilm-based vaccines are crucial for TB prevention.

Self-Assembled Borneol-Guanidine-Based Amphiphilic Polymers as an Efficient Antibiofilm Agent.

Biofilm-associated infections remain a tremendous obstacle to the treatment of microbial infections globally. However, the poor penetrability to a dense extracellular polymeric substance matrix of traditional antibacterial agents limits their antibiofilm activity. Here, we show that nanoaggregates formed by self-assembly of amphiphilic borneol-guanidine-based cationic polymers (BGNx-n) possess strong antibacterial activity and can eliminate mature Staphylococcus aureus (S. aureus) biofilms. The introduction of the guanidine moiety improves the hydrophilicity and membrane penetrability of BGNx-n. The self-assembled nanoaggregates with highly localized positive charges are expected to enhance their interaction with negatively charged bacteria and biofilms. Furthermore, nanoaggregates dissociate on the surface of biofilms into smaller BGNx-n polymers, which enhances their ability to penetrate biofilms. BGNx-n nanoaggregates that exhibit superior antibacterial activity have the minimum inhibitory concentration (MIC) of 62.5 μg·mL-1 against S. aureus and eradicate mature biofilms at 4 × MIC with negligible hemolysis. Taken together, this size-variable self-assembly system offers a promising strategy for the development of effective antibiofilm agents.

Optimizing chitosan derived from Metapenaeus affinis: a novel anti-biofilm agent against Pseudomonas aeruginosa

Pseudomonas aeruginosa is a commonly found Gram-negative bacterium in healthcare facilities and is renowned for its ability to form biofilms and its virulence factors that are controlled by quorum sensing (QS) systems. The increasing prevalence of multidrug-resistant strains of this bacterium poses a significant challenge in the field of medicine. Consequently, the exploration of novel antimicrobial agents has become a top priority. This research aims to optimize chitosan derived from white shrimp (Metapenaeus affinis) using the Response Surface Methodology (RSM) computational approach. The objective is to investigate chitosan’s potential as a solution for inhibiting QS activity and biofilm formation in P. aeruginosa ATCC 10,145. Under optimized conditions, chitin was treated with NaOH (1.41 M) for 15.75 h, HCl (7.49% vol) for 2.01 h, and at a deacetylation temperature of 81.15 °C. The resulting chitosan exhibited a degree of deacetylation (DD%) exceeding 93.98%, as confirmed by Fourier-transform infrared (FTIR) spectral analysis, indicating its high purity. The extracted chitosan demonstrated a significant synergistic antibiotic effect against P. aeruginosa when combined with ceftazidime, enhancing its bactericidal activity by up to 15-fold. In addition, sub-MIC (minimum inhibitory concentration) concentrations of extracted chitosan (10 and 100 µg/mL) successfully reduced the production of pyocyanin and rhamnolipid, as well as the swimming motility, protease activity and biofilm formation ability in comparison to the control group (P < 0.05). Moreover, chitosan treatment downregulated the RhlR and LasR genes in P. aeruginosa when compared to the control group (P < 0.05). The optimized chitosan extract shows significant potential as a coating agent for surgical equipment, effectively preventing nosocomial infections caused by P. aeruginosa pathogens.

Antibacterial and Antibiofilm Activity of Zinc Oxide Quantum Dots against Methicillin‐resistant Staphylococcus aureus

AbstractAntimicrobial resistance occurs mostly through the ineffective and unauthorized use of antibiotics in both the environment and health care. Biofilms are a new target in the search for new antibacterial agents. Here, the sol‐gel method is used to produce zinc oxide quantum dots (ZnO‐QDs). The ZnO‐QDs show yellow emission at 526 nm under 360 nm excitation. After that, the ability of ZnO‐QDs as an antibacterial and antibiofilm agent against methicillin‐resistant Staphylococcus aureus (MRSA) is examined. The 100 samples are collected from patients with hospital‐acquired infections between December 2020 and October 2021 from Imamen Kadhmiyan Teaching Hospital in Baghdad (Iraq). Out of these 100 samples, 63 are S. aureus and 20 out of 63 are MRSA. The minimum inhibitory concentration assay demonstrates the antibacterial activity of ZnO‐QDs on MRSA strains. Also, results show that 80, 5, and 15% of isolated MRSA strains produce strong, moderate, and weak or very weak biofilm, respectively. The effect of ZnO‐QDs on strong and moderate biofilms reveal that QDs could also inhibit and destroy biofilm produced by MRSA. In addition, the antibiofilm effect of ZnO‐QDs is greater than their antibacterial effect. Generally, this work offers new insights into the development of antibacterial and antibiofilm nanomaterials.

Enhanced applications in dentistry through autoclave-assisted sonochemical synthesis of Pb/Ag/Cu trimetallic nanocomposites

In recent years, researchers have increasingly focused on the development of multiphase trimetallic nanocomposites (TMNC) incorporating ternary metals or metal oxides, which hold significant potential as alternatives for combatting biofilms and bacterial infections. Enhanced oral health is ensured by the innovative techniques used to effectively prevent bacterial adherence and formation of biofilm on dental sutures. In this investigation, TMNC, which consists of Pb, Ag, and Cu, was synthesized using an autoclave-assisted sonochemical technique. Following synthesis, TMNC were characterized using FTIR, XRD, BET, XPS, TGA, and Raman spectroscopy to analyze their shape and microstructure. Subsequent evaluations, including MTT assay, antibacterial activity testing, and biofilm formation analysis, were conducted to assess the efficiency of the synthesized TMNC. Cytotoxicity and anti-human oral squamous cell carcinoma activities of TMNC were evaluated using the Human Oral Cancer cell line (KB) cell line through MTT assay, demonstrating a dose-dependent increase in anti-human oral squamous cell carcinoma activity against the KB cell line compared to the normal cell line, resulting in notably high cell viability. Furthermore, an ultrasonic probe was employed to incorporate TMNC onto dental suturing threads, with different concentrations of TMNC, ultrasonic power levels, and durations considered to determine optimal embedding conditions that result in the highest antibacterial activity. The inhibitory effects of TMNC, both in well diffusion assays and when incorporated into dental suturing threads, against gram-positive (Staphylococcus aureus) and gram-negative (Escherichia coli) bacteria on Mueller-Hinton agar (MHA) were assessed using various concentrations of TMNC. The results of the study indicated that the efficacy of TMNC in inhibiting bacterial growth on dental suturing threads remained impressive, even at low concentrations. Moreover, an evaluation of their potential to destabilize biofilms formed by S. aureus and E. coli, the two pathogens in humans, indicated that TMNC would be a promising anti-biofilm agent.

Nanoparticles of Cs0.33WO3 as Antibiofilm Agents and Photothermal Treatment to Inhibit Biofilm Formation.

Metal oxide nanoparticles with photothermal properties have attracted considerable research attention for their use in biomedical applications. Cesium tungsten oxide (Cs0.33WO3) nanoparticles (NPs) exhibit strong absorption in the NIR region due to localized surface plasmon resonance, through which they convert light to heat; hence, they can be applied to photothermal treatment for bacteria and biofilm ablation. Herein, Cs0.33WO3 NPs were synthesized through solid-phase synthesis, and their physical properties were characterized through Zetasizer, energy dispersive X-ray spectroscopy, Fourier transform infrared spectrometer, and scanning and transmission electron microscopy (SEM and TEM, respectively). Burkholderia cenocepacia isolates were cultured in tryptic soy broth supplemented with glucose, and the biofilm inhibition and antibiofilm effects of the NPs were determined using a crystal violet assay and the Cell Counting Kit-8 (CCK-8) assay. The biofilm morphology and viability of NP-treated cultures after NIR irradiation were evaluated through SEM and confocal microscopy, respectively. The cytotoxicity of NPs to human macrophages was also assessed using the CCK-8 assay. The NPs effectively inhibited biofilm formation, with a formation rate of <10% and a viability rate of <50% at the concentration of ≥200 μg/mL. The confocal analysis revealed that NIR irradiation markedly enhanced biofilm cytotoxicity after treatment with the NPs. The assay of cytotoxicity to human macrophages demonstrated the biocompatibility of the NPs and NIR irradiation. In sum, the Cs0.33WO3 NPs displayed effective biofilm inhibition and antibiofilm activity at 200 μg/mL treatment concentration; they exhibited an enhancement effect under the NIR irradiation, suggesting Cs0.33WO3 NPs are a potential candidate agent for NIR-irradiated photothermal treatment in bacterial biofilm inhibition and antibiofilm.

Ice nucleation active bacteria metabolites as antibiofilm agent to control Aeromonas hydrophila and Streptococcus agalactiae infections in Aquaculture

ObjectivesThe aim of this study was to quantify and identify metabolites of Ice Nucleation Active (INA) bacteria as an anti-biofilm agent against biofilms of fish pathogens such as Aeromonas hydrophila and Streptococcus agalactiae.ResultsIce nucleation active bacteria, which have the ability to catalyze ice nucleation, isolated from rainwater in previous studies, were used. All INA isolates were tested in several assays, including the antimicrobial test, which uses streptomycin as the positive control and none of the isolates were found positive in the antimicrobial test. As for the quorum quenching assay, it was found that four out of ten isolates were able to disturb the communication system in Chromobacterium violaceum wild type, which was used as the indicator bacteria. On the next assay, all ten isolates were tested for Biofilm Inhibition and Destruction and showed anti-biofilm activity with the highest percentage inhibition of 33.49% by isolate A40 against A. hydrophila and 77.26% by isolate A19 against S. agalactiae. C1 performed the highest destruction against A. hydrophila and S. agalactiae, with percentages of 32.11% and 51.88%, respectively. As for the GC-MS analysis, supernatants of INA bacteria contain bioactive compounds such as sarcosine and fatty acids, which are known to have antibiofilm activity against several biofilm-forming bacteria. Through 16s rRNA sequencing, identified bacteria are from the Pantoea, Enterobacter, and Acinetobacter genera. As for the conclusion, ice nucleation active bacteria metabolites tested showed positive results against pathogenic bacteria Aeromonas hydrophila and Streptococcus agalactiae in destructing and inhibiting biofilm growth.

Antibiofilm activity of d-limonene against spoilage Bacillus amyloliquefaciens

This study aimed to determine the efficacy of d-limonene in preventing biofilm formation by Bacillus amyloliquefaciens, a soy food spoilage bacterium. The minimal biofilm inhibitory concentrations (MBICs) of d-limonene against two strains of B. amyloliquefaciens were determined using a twofold dilution method. Furthermore, the ability of d-limonene to eradicate preformed biofilms was evaluated using the TTC reduction method. Subsequent analyses investigated the effects of d-limonene on biofilm morphology, cell motility, cell surface characteristics, and the composition of the biofilm matrix. The MBICs of d-limonene for B. amyloliquefaciens strains DY1a and DY1b were determined to be 24 and 36 μL/mL, respectively. At 2 MBIC, d-limonene eradicated 63.67% of the DY1a biofilms and 52.63% of the DY1b biofilms. Significantly, d-limonene markedly reduced swimming and swarming motility at concentrations ranging from 1/2 to 1 MBIC. In particular, at 1/2 MBIC, d-limonene decreased swimming and swarming abilities by 85.88% and 37.84%, respectively, for strain DY1a and by 75.00% and 95.07%, respectively, for strain DY1b. Furthermore, d-limonene suppressed the biosynthesis of polysaccharides and proteins in the extracellular biofilm matrix and reduced cell surface hydrophobicity, surface charge, and autoaggregation. These findings suggest that d-limonene could be an effective antibiofilm agent for the control of B. amyloliquefaciens in the soybean food industry.

Create artilysins from a recombinant library to serve as bactericidal and antibiofilm agents targeting Pseudomonas aeruginosa

Pseudomonas aeruginosa is a critical pathogen and novel treatments are urgently needed. The out membrane of P. aeruginosa facilitates biofilm formation and antibiotic resistance, and hinders the exogenous application against Gram-negative bacteria of endolysins. Engineered endolysins are investigated for enhancing antimicrobial activity, exemplified by artilysins. Nevertheless, existing research predominantly relies on laborious and time-consuming approaches of individually artilysin identification. This study proposes a novel strategy for expedited artilysin discovery using a recombinant artilysin library comprising proteins derived from 38 antimicrobial peptides and 8 endolysins. In this library, 19 colonies exhibited growth inhibition against P. aeruginosa exceeding 50 %, and three colonies were designated as dutarlysin-1, dutarlysin-2 and dutarlysin-3. Remarkably, dutarlysin-1, dutarlysin-2 and dutarlysin-3 demonstrated rapid and enhanced antibacterial activity, even minimum inhibitory concentration of them killed approximately 4.93 lg units, 6.75 lg units and 5.36 lg units P. aeruginosa, respectively. Dutarlysins were highly refractory to P. aeruginosa resistance development. Furthermore, 2 μmol/L dutarlysin-1 and dutarlysin-3 effectively eradicated over 76 % of the mature biofilm. These dutarlysins exhibited potential broad-spectrum activity against hospital susceptible Gram-negative bacteria. These results supported the effectiveness of this artilysins discovery strategy and suggested dutarlysin-1 and dutarlysin-3 could be promising antimicrobial agents for combating P. aeruginosa.

Diclofenac sodium effectively inhibits the biofilm formation of Staphylococcus epidermidis.

Staphylococcus epidermidis is an opportunistic pathogen commonly implicated in medical device-related infections. Its propensity to form biofilms not only leads to chronic infections but also exacerbates the issue of antibiotic resistance, necessitating high-dose antimicrobial treatments. In this study, we explored the use of diclofenac sodium, a non-steroidal anti-inflammatory drug, as an anti-biofilm agent against S. epidermidis. In this study, crystal violet staining and confocal laser scanning microscope analysis showed that diclofenac sodium, at subinhibitory concentration (0.4 mM), significantly inhibited biofilm formation in both methicillin-susceptible and methicillin-resistant S. epidermidis isolates. MTT assays demonstrated that 0.4 mM diclofenac sodium reduced the metabolic activity of biofilms by 25.21-49.01% compared to untreated controls. Additionally, the treatment of diclofenac sodium resulted in a significant decrease (56.01-65.67%) in initial bacterial adhesion, a crucial early phase of biofilm development. Notably, diclofenac sodium decreased the production of polysaccharide intercellular adhesin (PIA), a key component of the S. epidermidis biofilm matrix, in a dose-dependent manner. Real-time quantitative PCR analysis revealed that diclofenac sodium treatment downregulated biofilm-associated genes icaA, fnbA, and sigB and upregulated negative regulatory genes icaR and luxS, providing potential mechanistic insights. These findings indicate that diclofenac sodium inhibits S. epidermidis biofilm formation by affecting initial bacterial adhesion and the PIA synthesis. This underscores the potential of diclofenac sodium as a supplementary antimicrobial agent in combating staphylococcal biofilm-associated infections.

Essential oils of indigenous citrus varieties of Northeast India as potential antibiofilm agents against foodborne pathogens: An in vitro and in silico study

Essential oils of indigenous citrus varieties of Northeast India as potential antibiofilm agents against foodborne pathogens: An in vitro and in silico study