Epidermidis Biofilm Research Articles

Staphylococcus epidermidis biofilms undergo metabolic and matrix remodeling under nitrosative stress.

Staphylococcus epidermidis is a commensal skin bacterium that forms host- and antibiotic-resistant biofilms that are a major cause of implant-associated infections. Most research has focused on studying the responses to host-imposed stresses on planktonic bacteria. In this work, we addressed the open question of how S. epidermidis thrives on toxic concentrations of nitric oxide (NO) produced by host innate immune cells during biofilm assembly. We analyzed alterations of gene expression, metabolism, and matrix structure of biofilms of two clinical isolates of S. epidermidis, namely, 1457 and RP62A, formed under NO stress conditions. In both strains, NO lowers the amount of biofilm mass and causes increased production of lactate and decreased acetate excretion from biofilm glucose metabolism. Transcriptional analysis revealed that NO induces icaA, which is directly involved in polysaccharide intercellular adhesion (PIA) production, and genes encoding proteins of the amino sugar pathway (glmM and glmU) that link glycolysis to PIA synthesis. However, the strains seem to have distinct regulatory mechanisms to boost lactate production, as NO causes a substantial upregulation of ldh gene in strain RP62A but not in strain 1457. The analysis of the matrix components of the staphylococcal biofilms, assessed by confocal laser scanning microscopy (CLSM), showed that NO stimulates PIA and protein production and interferes with biofilm structure in a strain-dependent manner, but independently of the Ldh level. Thus, NO resistance is attained by remodeling the staphylococcal matrix architecture and adaptation of main metabolic processes, likely providing in vivo fitness of S. epidermidis biofilms contacting NO-proficient macrophages.

Antibacterial activities of water extract of Vernonia amygdalina Delile leaves

Staphylococci are common bacterial colonizers of the skin and mucous membranes of humans and other mammals. Staphylococcus epidermidis in particular is the most frequently isolated species from human epithelia. It colonizes predominantly the axillae, head, and nares. One type of medicinal plant that is nutritious for health is African leaves (Vernonia amygdalina Delile). Vernonia amygdalina extract showed the presence of some compounds which are rich in those constituents. The phytochemical screening of the extracts showed variation in their phytochemical constituents with the presence and or absence of some components. The presence of glycosides, alkaloids, and flavonoids was believed to exhibit the antibiotic properties of V. amygdalina leaves and confirmed their antimicrobial efficacy against selected pathogens. The concentration of 50 mg/mL up to a concentration of 1.56 mg/mL can inhibit the growth of S. epidermidis, the Minimum Inhibitory Concentration (MIC) itself is at the smallest concentration that has been able to inhibit bacterial growth and the number of bacteria is < 10 colonies. Staphylococcus epidermidis at a concentration of 25 mg/mL, absorbance values at wavelengths of 260 nm and 280 nm were higher than the absorbance values of S. epidermidis at a concentration of 12.5 mg/mL. The absorbance value at wavelengths of 260 nm and 280 nm will increase with the higher concentration of the extract, this indicates the presence of cell leakage observed in the presence of protein and nucleic acid leakage. S. epidermidis biofilm inhibition test shows that the results of the absorbance or OD (Optical Density) measurement in the biofilm inhibition test using the ethanol extract of V. amygdalina Delile, the OD value at a concentration of 25 mg/mL ie 0.149±0.004 showed a biofilm inhibition result of 62.37% greater than the concentration of 12.5 mg/mL which is 0.318±0.003 with a biofilm inhibition result of 19.52%, this indicates that the higher the concentration of the extract, the smaller the biofilm formation of S. epidermidis bacteria.

In vitro and in vivo evaluation of diethyldithiocarbamate with copper ions and its liposomal formulation for the treatment of Staphylococcus aureus and Staphylococcus epidermidis biofilms

Surgical site infections (SSIs) are mainly caused by Staphylococcus aureus (S. aureus) and Staphylococcus epidermidis (S. epidermidis) biofilms. Biofilms are aggregates of bacteria embedded in a self-produced matrix that offers protection against antibiotics and promotes the spread of antibiotic-resistance in bacteria. Consequently, antibiotic treatment frequently fails, resulting in the need for alternative therapies. The present study describes the in vitro efficacy of the Cu(DDC)2 complex (2:1 M ratio of diethyldithiocarbamate (DDC−) and Cu2+) with additional Cu2+ against S. aureus and S. epidermidis biofilms in models mimicking SSIs and in vitro antibacterial activity of a liposomal Cu(DDC)2 + Cu2+ formulation. The in vitro activity on S. aureus and S. epidermidis biofilms grown on two hernia mesh materials and in a wound model was determined by colony forming unit (CFU) counting. Cu2+-liposomes and Cu(DDC)2-liposomes were prepared, and their antibacterial activity was assessed in vitro using the alamarBlue assay and CFU counting and in vivo using a Galleria mellonella infection model. The combination of 35 μM DDC− and 128 μM Cu2+ inhibited S. aureus and S. epidermidis biofilms on meshes and in a wound infection model. Cu(DDC)2-liposomes + free Cu2+ displayed similar antibiofilm activity to free Cu(DDC)2 + Cu2+, and significantly increased the survival of S. epidermidis-infected larvae. Whilst Cu(DDC)2 + Cu2+ showed substantial antibiofilm activity in vitro against clinically relevant biofilms, its application in mammalian in vivo models is limited by solubility. The liposomal Cu(DDC)2 + Cu2+ formulation showed antibiofilm activity in vitro and antibacterial activity and low toxicity in G. mellonella, making it a suitable water-soluble formulation for future application on infected wounds in animal trials.

Surfaces modified with small molecules that interfere with nucleotide signaling reduce Staphylococcus epidermidis biofilm and increase the efficacy of ciprofloxacin

Staphylococcus epidermidis are common bacteria associated with biofilm related infections on implanted medical devices. Antibiotics are often used in combating such infections, but they may lose their efficacy in the presence of biofilms. Bacterial intracellular nucleotide second messenger signaling plays an important role in biofilm formation, and interference with the nucleotide signaling pathways provides a possible way to control biofilm formation and to increase biofilm susceptibility to antibiotic therapy. This study synthesized small molecule derivates of 4-arylazo-3,5-diamino-1 H-pyrazole (named as SP02 and SP03) and found these molecules inhibited S. epidermidis biofilm formation and induced biofilm dispersal. Analysis of bacterial nucleotide signaling molecules showed that both SP02 and SP03 significantly reduced cyclic dimeric adenosine monophosphate (c-di-AMP) levels in S. epidermidis at doses as low as 25 µM while having significant effects on multiple nucleotides signaling including cyclic dimeric guanosine monophosphate (c-di-GMP), c-di-AMP, and cyclic adenosine monophosphate (cAMP) at high doses (100 µM or greater). We then tethered these small molecules to polyurethane (PU) biomaterial surfaces and investigated biofilm formation on the modified surfaces. Results showed that the modified surfaces significantly inhibited biofilm formation during 24 h and 7-day incubations. The antibiotic ciprofloxacin was used to treat these biofilms and the efficacy of the antibiotic (2 µg/mL) was found to increase from 94.8% on unmodified PU surfaces to > 99.9% on both SP02 and SP03 modified surfaces (>3 log units). Results demonstrated the feasibility of tethering small molecules that interfere with nucleotide signaling onto polymeric biomaterial surfaces and in a way that interrupts biofilm formation and increases antibiotic efficacy for S. epidermidis infections.

Eradication of Staphylococcus epidermidis within Biofilms: Comparison of Systemic versus Supratherapeutic Concentrations of Antibiotics.

Biofilm-forming bacterial infections result in clinical failure, recurring infections, and high health care costs. The antibiotic concentrations needed to eradicate biofilm require further research. We aimed to model an in vitro prosthetic joint infection (PJI) to elucidate the activity of traditional systemic concentrations versus supratherapeutic concentrations to eradicate a Staphylococcus epidermidis biofilm PJI. We evaluated S. epidermidis high-biofilm-forming (ATCC 35984) and low-biofilm-forming (ATCC 12228) isolates in an in vitro pharmacodynamic biofilm reactor model with chromium cobalt coupons to simulate prosthetic joint infection. Vancomycin, daptomycin, levofloxacin, and minocycline were used alone and combined with rifampin to evaluate the effect of biofilm eradication. We simulated three exposures: (i) humanized systemic dosing alone, (ii) supratherapeutic doses (1,000× MIC), and (iii) and dosing in combination with rifampin. Resistance development was monitored throughout the study. Simulated humanized systemic doses of a lipoglycopeptide (daptomycin), a fluoroquinolone (levofloxacin), a tetracycline (minocycline), and a glycopeptide (vancomycin) alone failed to eradicate a formed S. epidermidis biofilm. Supratherapeutic doses of vancomycin (2,000 μg/mL) and minocycline (15 μg/mL) with or without rifampin (15 μg/mL) failed to eradicate biofilms. However, a levofloxacin supratherapeutic dose (125 μg/mL) with rifampin eradicated the high-biofilm-producing isolate by 48 h. Interestingly, supratherapeutic-dose exposures of daptomycin (500 μg/mL) alone eradicated high- and low-biofilm-forming isolates in established biofilms. The concentrations needed to eradicate biofilms on foreign materials are not obtained with systemic dosing regimens. The failure of systemic dosing regimens to eradicate biofilms validates clinical findings with recurring infections. The addition of rifampin to supratherapeutic dosing regimens does not result in synergy. Supratherapeutic daptomycin dosing may be effective at the site of action to eradicate biofilms. Further studies are needed.

ROUTINE SURGICAL ANTISEPTIC AGENTS ARE INEFFECTIVE IN THE ERADICATION OF TYPICAL PJI BIOFILM FORMED ON ORTHOPAEDIC IMPLANT MATERIAL IN VITRO AND THE ABSENCE OF AN ANTISEPTIC NEUTRALIZATION STAGE GIVES THE FALSE IMPRESSION OF EFFICACY

Prosthetic joint infection (PJI) is a serious complication following joint replacement. Antiseptic solutions are often used for intraoperative wound irrigation particularly in cases of revision for PJI. Antiseptic irrigation is intended to eradicate residual bacteria which may be either free floating or in residual biofilm although there is no clear clinical efficacy for its use. Also, reviewing the scientific literature there is discordance in in vitro results where some studies questions antiseptic efficacy whilst others suggest that even at low concentration antiseptic agents are effective at eradicating bacterial biofilms.The aim of this in vitro study was to establish the efficacy of undiluted antiseptic agents at eradication of a typical PJI forming biofilm and determine the importance of an antiseptic neutralisation step in this assessment.Mature Staphylococcus epidermidis biofilms grown on TiAl6V4 discs were submerged in chlorohexidine (CHL) gluconate 4%, povidone-iodine (PI) 10% or phosphate-buffered saline (PBS) control solution. The discs were then rinsed, the biofilm bacteria suspended in solution using sonication and vortexing, and the viable count (CFU/ml) of the bacterial suspensions determined. The rinse/suspension solution was either (a) PBS or (b) Dey-Engley neutralization broth (NB).When PBS was used to rinse/suspend the biofilm a highly significant, 7.5 and 4.1, mean log reduction in biofilm vitality was observed from the control, for CHL 4% and PI 10%, respectively. However, when NB was the rinse/suspension solution the apparent antiseptic biofilm eradication efficacy was replaced with a statistically significant but clinically irrelevant less the one log-reduction in biofilm vitality.Clinical antiseptic agents are ineffective at eradicating S. epidermidis biofilm in an in vitro PJI model and absence of a neutralisation step gives the false impression of efficacy. Antiseptics alone are an ineffective treatment for biofilm related PJI and no substitute for meticulous debridement.

Assessment of the antibacterial effect of Barium Titanate nanoparticles against Staphylococcus epidermidisadhesion after addition to maxillofacial silicone.

Background: Maxillofacial silicones are the most popular and acceptable material for making maxillofacial prostheses, but they are not perfect in every sense. To enhance their effectiveness, more improvements to their properties are required, such as their antimicrobial efficiency. This study assess the antibacterial effect of barium titanate nanoparticles in various percentages against staphylococcus epidermidis biofilm adhesion after addition to maxillofacial silicone. Methods: Barium titanate nanoparticles were added into VST-50 platinum silicone elastomer in four weight percentages (0.25wt%, 0.5wt%, 0.75wt% and 1wt%). 50 specimens were prepared and categorized into five groups; one control group and four experimental groups. All conducted data was statistically analyzed using (one-way ANOVA) analysis of variance, and Games-Howell multiple comparison test (significant level at p < 0.05). Shapiro-Wilk and Levene's tests were used, respectively, to evaluate the normal distribution and homogeneity of the data. Result: One-way ANOVA test revealed a highly significant difference between all groups, and Games-Howell test revealed a highly significant difference between the control group and the four experimental groups. The 0.25wt% and 0.5wt% groups revealed a highly significant difference between them and with the (0.75%wt and 0.1%wt) groups. While the 0.75wt% group revealed a significant difference with 1wt% group. Conclusions: The addition of barium titanate to VST-50 maxillofacial silicone enhanced the antibacterial activity of silicon against Staphylococcus epidermidis, and this activity seems to be concentration dependent. FTIR analysis demonstrated no chemical interaction between the Barium Titanate and the VST-50 maxillofacial silicone elastomer. SEM pictures show that the barium titanate nanopowder was effectively dispersed inside the maxillofacial silicone matrix.

Rejuvenating the [1, 2, 3]-triazolo [1,5-a]quinoxalin-4(5H)-one scaffold: Synthesis and derivatization in a sustainable guise and preliminary antimicrobial evaluation.

The [1,2,3]-triazolo [1,5-a] quinoxalin-4(5H)-one scaffold and its analogues triazole-fused heterocyclic compounds are relevant structural templates in both natural and synthetic biologically active compounds. However, their medicinal chemistry applications are often limited due to the lack of synthetic protocols combining straightforward generation of the central core while also allowing extensive decoration activity for drug discovery purposes. Herein, we report a "refreshed" synthesis of the [1,2,3]-triazolo [1,5-a]quinoxalin-4(5H)-one core, encompassing the use of eco-compatible catalysts and reaction conditions. We have also performed a sustainable and extensive derivatization campaign at both the endocyclic amide nitrogen and the ester functionality, comprehensively exploring the reaction scope and overcoming some of the previously reported difficulties in introducing functional groups on this structural template. Finally, we unveiled a preliminary biological investigation for the newly generated chemical entities. Our assessment of the compounds on different bacterial species (two S. aureus strains, three P. aeruginosa strains, K. pneumonia), and two fungal C. albicans strains, as well as the evaluation of their activity on S. epidermidis biofilm formation, foster further optimization for the retrieved hit compounds 9, 14, and 20.

Design, Synthesis, and Evaluation of Carbonate-Linked Halogenated Phenazine-Quinone Prodrugs with Improved Water-Solubility and Potent Antibacterial Profiles.

Pathogenic bacteria have devastating impacts on human health as a result of acquired antibiotic resistance and innate tolerance. Every class of our current antibiotic arsenal was initially discovered as growth-inhibiting agents that target actively replicating (individual, free-floating) planktonic bacteria. Bacteria are notorious for utilizing a diversity of resistance mechanisms to overcome the action of conventional antibiotic therapies and forming surface-attached biofilm communities enriched in (non-replicating) persister cells. To address problems associated with pathogenic bacteria, our group is developing halogenated phenazine (HP) molecules that demonstrate potent antibacterial and biofilm-eradicating activities through a unique iron starvation mode of action. In this study, we designed, synthesized, and investigated a focused collection of carbonate-linked HP prodrugs bearing a quinone trigger to target the reductive cytoplasm of bacteria for bioactivation and subsequent HP release. The quinone moiety also contains a polyethylene glycol group, which dramatically enhances the water-solubility properties of the HP-quinone prodrugs reported herein. We found carbonate-linked HP-quinone prodrugs 11, 21-23 to demonstrate good linker stability, rapid release of the active HP warhead following dithiothreitol (reductive) treatment, and potent antibacterial activities against methicillin-resistant Staphylococcus aureus (MRSA), methicillin-resistant Staphylococcus epidermidis, and Enterococcus faecalis. In addition, HP-quinone prodrug 21 induced rapid iron starvation in MRSA and S. epidermidis biofilms, illustrating prodrug action within these surface-attached communities. Overall, we are highly encouraged by these findings and believe that HP prodrugs have the potential to address antibiotic resistant and tolerant bacterial infections.

A multi-targeting bionanomatrix coating to reduce capsular contracture development on silicone implants

BackgroundCapsular contracture is a critical complication of silicone implantation caused by fibrotic tissue formation from excessive foreign body responses. Various approaches have been applied, but targeting the mechanisms of capsule formation has not been completely solved. Myofibroblast differentiation through the transforming growth factor beta (TGF-β)/p-SMADs signaling is one of the key factors for capsular contracture development. In addition, biofilm formation on implants may result chronic inflammation promoting capsular fibrosis formation with subsequent contraction. To date, there have been no approaches targeting multi-facted mechanisms of capsular contracture development.MethodsIn this study, we developed a multi-targeting nitric oxide (NO) releasing bionanomatrix coating to reduce capsular contracture formation by targeting myofibroblast differentiation, inflammatory responses, and infections. First, we characterized the bionanomatrix coating on silicon implants by conducting rheology test, scanning electron microcsopy analysis, nanoindentation analysis, and NO release kinetics evaluation. In addition, differentiated monocyte adhesion and S. epidermidis biofilm formation on bionanomatrix coated silicone implants were evaluated in vitro. Bionanomatrix coated silicone and uncoated silicone groups were subcutaneously implanted into a mouse model for evaluation of capsular contracture development for a month. Fibrosis formation, capsule thickness, TGF-β/SMAD 2/3 signaling cascade, NO production, and inflammatory cytokine production were evaluated using histology, immunofluorescent imaging analysis, and gene and protein expression assays.ResultsThe bionanomatrix coating maintained a uniform and smooth surface on the silicone even after mechanical stress conditions. In addition, the bionanomatrix coating showed sustained NO release for at least one month and reduction of differentiated monocyte adhesion and S. epidermidis biofilm formation on the silicone implants in vitro. In in vivo implantation studies, the bionanomatrix coated groups demonstrated significant reduction of capsule thickness surrounding the implants. This result was due to a decrease of myofibroblast differentiation and fibrous extracellular matrix production through inhibition of the TGF-β/p-SMADs signaling. Also, the bionanomatrix coated groups reduced gene expression of M1 macrophage markers and promoted M2 macrophage markers which indicated the bionanomatrix could reduce inflammation but promote healing process.ConclusionsIn conclusion, the bionanomatrix coating significantly reduced capsular contracture formation and promoted healing process on silicone implants by reducing myfibroblast differentiation, fibrotic tissue formation, and inflammation.Graphical A multi-targeting nitric oxide releasing bionanomatrix coating for silicone implant can reduce capsular contracture and improve healing process. The bionanomatrix coating reduces capsule thickness, α-smooth muscle actin and collagen synthesis, and myofibroblast differentiation through inhibition of TGF-β/SMADs signaling cascades in the subcutaneous mouse models for a month.

Do Lactobacillus rhamnosus-Originated Probiotic and Parabiotic Have Inhibitory Effects on Intraocular Lens Biofilm?

Objective: Our aim was to investigate the formation of Staphylococcus (S.) epidermidis biofilm on hydrophobic acrylic lenses and whether the inhibition of the formed biofilm is possible with probiotic Lactobacillus (L.) rhamnosus 312 and parabiotic prepared from it. Materials and Methods: The probiotic bacteria L. rhamnosus 312 and intercellular adhesion (ICA) gene-positive tested bacteria S. epidermidis KA15.8 were used in the study from stock. To obtain the parabiotic the cultures were developed in De Man Rogosa and Sharpe (MRS) broth for 48 hours and autoclaved at 121ºC for 15 minutes. Biofilms on hydrophobic acrylic intraocular lenses and the antibiofilm effects of parabiotic and probiotic L. rhamnosus were evaluated. Scanning electron microscopy photos of biofilms produced on intraocular lenses (IOLs) were taken. Results: Probiotic L. rhamnosus 312 and the parabiotic test showed antibacterial activity on test bacteria, ICA positive S. epidermidis KA15.8. However, the probiotic L. rhamnosus 312 zone diameter was found to be wider. After the biofilm was formed, the addition of parabiotic inhibited the biofilm formed by S. epidermis KA15.8 by 58.29%. The number of S. epidermidis KA15.8 in the biofilm also decreased. Conclusion: Parabiotic and probiotic L. rhamnosus 312 was found effective for its antibiofilm effect. However, further studies with different concentrations are needed.

Cec4-Derived Peptide Inhibits Planktonic and Biofilm-Associated Methicillin Resistant Staphylococcus epidermidis.

Staphylococcus epidermidis is part of the normal microbiota that colonizes the skin and mucosal surfaces of human beings. Previous studies suggested that S. epidermidis possessed low virulence, but recent studies confirmed that it can acquire high virulence from Staphylococcus aureus and with the increasing detection of methicillin-resistant S. epidermidis. It has become a major pathogen of graft-associated and hospital-acquired infections. In previous studies, we modified the antimicrobial peptide Cec4 (41 amino acids) and obtained the derived peptide C9 (16 amino acids) showing better antimicrobial activity against S. epidermidis with an MIC value of 8 μg/mL. The peptide has rapid bactericidal activity without detectable high-level resistance, showing certain inhibition and eradication ability on S. epidermidis biofilms. The damage of cell membrane structures by C9 was observed by scanning emission microscopy (SEM) and transmission electron microscopy (TEM). In addition, C9 altered the S. epidermidis cell membrane permeability, depolarization levels, fluidity, and reactive oxygen species (ROS) accumulation and possessed the ability to bind genomic DNA. Analysis of the transcriptional profiles of C9-treated cells revealed changes in genes involved in cell wall and ribosome biosynthesis, membrane protein transport, oxidative stress, and DNA transcription regulation. At the same time, the median lethal dose of C9 in mice was more than 128 mg/kg, and the intraperitoneal administration of 64 mg/kg was less toxic to the liver and kidneys of mice. Furthermore, C9 also showed a certain therapeutic effect on the mouse bacteremia model. In conclusion, C9 may be a candidate drug against S. epidermidis, which has the potential to be further developed as an antibacterial therapeutic agent. IMPORTANCE S. epidermidis is one of the most important pathogens of graft-related infection and hospital-acquired infection. The growing problem of antibiotic resistance, as well as the emergence of bacterial pathogenicity, highlights the need for antimicrobials with new modes of action. Antimicrobial peptides have been extensively studied over the past 30 years as ideal alternatives to antibiotics, and we report here that the derived peptide C9 is characterized by rapid bactericidal and antibiofilm activity, avoiding the development of resistance by acting on multiple nonspecific targets of the cell membrane or cell components. In addition, it has therapeutic potential against S. epidermidis infection in vivo. This study provides a rationale for the further development and application of C9 as an effective candidate antibiotic.

CATASAN Is a New Anti-Biofilm Agent Produced by the Marine Antarctic Bacterium Psychrobacter sp. TAE2020.

The development of new approaches to prevent microbial surface adhesion and biofilm formation is an emerging need following the growing understanding of the impact of biofilm-related infections on human health. Staphylococcus epidermidis, with its ability to form biofilm and colonize biomaterials, represents the most frequent causative agent involved in infections of medical devices. In the research of new anti-biofilm agents against S. epidermidis biofilm, Antarctic marine bacteria represent an untapped reservoir of biodiversity. In the present study, the attention was focused on Psychrobacter sp. TAE2020, an Antarctic marine bacterium that produces molecules able to impair the initial attachment of S. epidermidis strains to the polystyrene surface. The setup of suitable purification protocols allowed the identification by NMR spectroscopy and LC-MS/MS analysis of a protein-polysaccharide complex named CATASAN. This complex proved to be a very effective anti-biofilm agent. Indeed, it not only interferes with cell surface attachment, but also prevents biofilm formation and affects the mature biofilm matrix structure of S. epidermidis. Moreover, CATASAN is endowed with a good emulsification activity in a wide range of pH and temperature. Therefore, its use can be easily extended to different biotechnological applications.

Effect Of Shikimic Acid And Acetylsalicylic Acid On Biofilm Formation In Staphylococcus Epidermidis

The aim of the current study was to evaluate the activity of Shikimic acid (SA) and Acetylsalicylic acid (ASA) against S. epidermidis biofilms formation in different conditions. A total of (231) samples were collected from patients who attended different hospitals in Ramadi and Faluja cities during the period from December(2020) to the end of July (2021). Samples were cultured on blood, and manitol salt agar and identification was performed by the VITEK 2 system. Antibiofilm activity of SA was evaluated by two methods, modified crystal violet, and MTT assay. The SA was tested in five MIC (0.25 MIC, 0.5 MIC, MIC2, MIC, and 4 MIC) and was tested during different incubation periods (2 hr, 4 hr, 6 hr, 12 hr, and 24 hr), Shikimic acid gave highest percentage inhibition of biofilm formation was 93% at 12hr and ,94% at 24hr , while Acetyl salicylic acid was gave 85% at 12hr and 91% at 24hr incubation period . Then evaluated the synergistic activity of SA and ASA acid with ceftriaxone and azithromycin against biofilm formation also then determination of the antibiofilm activity of SA and ASA acid against bacteria adherence to Foley balloon latex catheter (FBLC) pieces by REMA assay.In addition, determine the efficacy of SA and ASA acid against cell adherence using murine mammary adenocarcinoma cell line (amn3)cells. The results showed high percentage inhibition of biofilm formation as the concentration of SA and ASA increased . In conclusion, SA and ASA have the potential to be further developed as a new antitumor agent due to their impressive antibiofilm action against bacteria. shikimic acid , acetyle salicylic acid were gave up regulation of atlE gene , 2 ( −ΔΔCT) was (2)for shikimic acid while (3)for acetyl salicylic acid , aap,icaA genes, all antibiofilm agents were gave down regulation result 2 ( −ΔΔCT) was (0.2)for shikimic acid while (0.1)for acetyl salicylic acid.

Ultrasonic preparation of Tween-essential oil (Zanthoxylum schinifolium Sieb. et Zucc) oil/water nanoemulsion: Improved stability and alleviation of Staphylococcus epidermidis biofilm

Staphylococcus epidermidis has been identified as a pathogen in ready-to-eat foods, and its biofilm can cause devastating chronic infections in affected hosts. For the demand of natural antibacterial reagents and overcoming their inherent properties, nanoemulsions are increasingly being used in the delivery of natural drugs. The nanoemulsion obtained by ultrasonic technology has the advantages of small droplet size , low polydispersity index (PDI), high stability and small amount of surfactant. Therefore, ultrasonic technology was used to prepare Zanthoxylum schinifolium Sieb. et Zucc essential oil nanoemulsion (ZEO-NE), a novel antibiofilm nanoemulsion with a mean particle size of 105.6 nm, polydispersity index of 0.278, and Zeta potential of − 22.3 mV. Assessment of antibacterial activity defined the minimum inhibitory concentration (MIC) and minimum bactericidal concentration of ZEO-NE against S. epidermidis as 2.5 and 5.0 mg/mL, respectively. Meanwhile, the antibiofilm assays revealed that the inhibition rate of ZEO-NE at sub-MIC levels (1/16–1/2 MIC) against S. epidermidis biofilm formation was 4.65–81.38 %. Further studies showed that sub-MIC ZEO-NE can effectively inhibit the formation of biofilms by extracellular polymeric substances and regulate gene expression of biofilm initial adhesion, formation, and quorum-sensing systems. Meanwhile, the inhibition rates of ZEO-NE (1/2–4 MIC) on preformed biofilm biomass and cell metabolic activity were 45.06–72.40 % and 30.72–62.54 %, respectively. In addition, electron microscopy analysis showed that ZEO-NE can destroy the established biofilms to a great extent. Finally, cytotoxicity evaluation experiments showed that ZEO-NE could damage human immortalized keratinocytes . These findings indicate that nanotechnology is an effective strategy to increase ZEO stability and solubility, and the prepared ZEO-NE is a viable candidate to inhibit S. epidermidis and its biofilms. • A high-quality ZEO-NE with a size of 105.6 nm, PDI of 0.278 and Zeta potential of − 22.3 mV was obtained. • The ZEO-NE can strongly inhibit Staphylococcus epidermidis and its biofilms. • The ZEO-NE can down-regulate the expression of virulence genes of S. epidermidis . • The ZEO-NE can effectively destroy the stable structure of the preformed biofilm.

The endoglycosidase activity of Dispersin B is mediated through electrostatic interactions with cationic poly-β-(1→6)-N-acetylglucosamine.

Bacterial biofilms consist of bacterial cells embedded within a self-produced extracellular polymeric substance (EPS) composed of exopolysaccharides, extra cellular DNA, proteins and lipids. The enzyme Dispersin B (DspB) is a CAZy type 20 β-hexosaminidase enzyme that catalyses the hydrolysis of poly-N-acetylglucosamine (PNAG), a major biofilm polysaccharide produced by a wide variety of biofilm-forming bacteria. Native PNAG is partially de-N-acetylated, and the degree of deacetylation varies between species and dependent on the environment. We have previously shown that DspB is able to perform both endo- and exo-glycosidic bond cleavage of PNAG depending on the de-N-acetylation patterns present in the PNAG substrate. Here, we used a combination of synthetic PNAG substrate analogues, site-directed mutagenesis and in vitro biofilm dispersal assay to investigate the molecular basis for the endo-glycosidic cleavage activity of DspB and the importance of this activity for dispersal of PNAG-dependent Staphylococcus epidermidis biofilms. We found that D242 contributes to the endoglycosidase activity of DspB through electrostatic interactions with cationic substrates in the -2 binding site. A DspBD242N mutant was highly deficient in endoglycosidase activity while maintaining exoglycosidase activity. When used to disperse S. epidermidis biofilms, this DspBD242N mutant resulted in an increase in residual biofilm biomass after treatment when compared to wild-type DspB. These results suggest that the de-N-acetylation of PNAG in S. epidermidis biofilms is not uniformly distributed and that the endoglycosidase activity of DspB is required for efficient biofilm dispersal.

Correlation notice on the electrochemical dealloying and antibacterial properties of gold-silver alloy nanoparticles.

Galvanic replacement reaction was used in the synthesis of bimetallic gold-silver alloy nanoparticles (Au-Ag NPs), where pre-synthesized Ag nanoparticles-polyvinylpyrrolidone (AgNPs-PVP) were used to reduce the aryldiazonium tetrachloroaurate(III) salt in water. TEM images and EDS elemental analysis showed the formation of spherical Au-Ag NPs with sizes of 12.8 ± 4.9nm and 25.6 ± 14.4nm for corresponding Au-Ag ratios and termed as Au0.91Ag0.09 and Au0.79Ag0.21, respectively, with different concentrations of the gold precursor. The hydrodynamic sizes measured using dynamic light scattering are 46.4nm and 74.8nm with corresponding zeta potentials of - 44.56 and - 25.09mV in water, for Au0.91Ag0.09 and Au0.79Ag0.21 respectively. Oxidative leachability of Ag ion studies from the starting AgNPs-PVP in 1M NaCl showed a significant decrease in the plasmon peak after 8h, indicating the complete dissolution of Ag ions, however, there is enhanced oxidation resistivity of Ag from Au-Ag NPs even after 24h. Electrochemical studies on glassy carbon electrodes displayed a low oxidation peak in aqueous solutions of 20mM KCl at 0.16V and KNO3 at 0.33V vs. saturated calomel electrode (SCE). We studied the antibacterial activity of Au-Ag alloy nanoparticles against gram-positive Staphylococcus aureus, Staphylococcus epidermidis, Enterococcusfaecalis, and gram-negative Escherichia coli, Salmonella typhimurium, and Pseudomonas aeruginosa. Our findings demonstrated superior antibacterial activity of Au-Ag NPs compared with AgNPs-PVP. Moreover, the nanoparticles inhibited the S. epidermidis biofilm formation.

Role of RNAIII in Resistance to Antibiotics and Antimicrobial Agents in Staphylococcus epidermidis Biofilms.

Staphylococcus epidermidis is a known opportunistic pathogen and is one of the leading causes of chronic biofilm-associated infections. Biofilm formation is considered as a main strategy to resist antibiotic treatment and help bacteria escape from the human immune system. Understanding the complex mechanisms in biofilm formation can help find new ways to treat resistant strains and lower the prevalence of nosocomial infections. In order to examine the role of RNAIII regulated by the agr quorum sensing system and to what extent it influences biofilm resistance to antimicrobial agents, deletion mutant S. epidermidis RP62a-ΔRNAIII deficient in repressor domains with a re-maining functional hld gene was created. A deletion strain was used to examine the influence of oxacillin in combination with vanillin on biofilm resistance and cell survival was determined. Utilizing real-time qPCR, confocal laser scanning microscopy (CLSM), and crystal violet staining analyses, we found that the RNAIII-independent controlled phenol soluble modulins (PSMs) and RNAIII effector molecule have a significant role in biofilm resistance to antibiotics and phenolic compounds, and it protects the integrity of biofilms. Moreover, a combination of antibiotic and antimicrobial agents can induce methicillin-resistant S. epidermidis biofilm formation and can lead to exceedingly difficult medical treatment.

The combination of diethyldithiocarbamate and copper ions is active against Staphylococcus aureus and Staphylococcus epidermidis biofilms in vitro and in vivo.

Staphylococcus aureus and Staphylococcus epidermidis are associated with life-threatening infections. Despite the best medical care, these infections frequently occur due to antibiotic resistance and the formation of biofilms of these two bacteria (i.e., clusters of bacteria embedded in a matrix). As a consequence, there is an urgent need for effective anti-biofilm treatments. Here, we describe the antibacterial properties of a combination treatment of diethyldithiocarbamate (DDC) and copper ions (Cu2+) and their low toxicity in vitro and in vivo. The antibacterial activity of DDC and Cu2+ was assessed in vitro against both planktonic and biofilm cultures of S. aureus and S. epidermidis using viability assays, microscopy, and attachment assays. Cytotoxicity of DDC and Cu2+ (DDC-Cu2+) was determined using a human fibroblast cell line. In vivo antimicrobial activity and toxicity were monitored in Galleria mellonella larvae. DDC-Cu2+ concentrations of 8 μg/ml DDC and 32 μg/ml Cu2+ resulted in over 80% MRSA and S. epidermidis biofilm killing, showed synergistic and additive effects in both planktonic and biofilm cultures of S. aureus and S. epidermidis, and synergized multiple antibiotics. DDC-Cu2+ inhibited MRSA and S. epidermidis attachment and biofilm formation in the xCELLigence and Bioflux systems. In vitro and in vivo toxicity of DDC, Cu2+ and DDC-Cu2+ resulted in > 70% fibroblast viability and > 90% G. mellonella survival. Treatment with DDC-Cu2+ significantly increased the survival of infected larvae (87% survival of infected, treated larvae vs. 47% survival of infected, untreated larvae, p < 0.001). Therefore, DDC-Cu2+ is a promising new antimicrobial with activity against planktonic and biofilm cultures of S. epidermidis and S. aureus and low cytotoxicity in vitro. This gives us high confidence to progress to mammalian animal studies, testing the antimicrobial efficacy and safety of DDC-Cu2+.

What is the best technic to dislodge Staphylococcus epidermidis biofilm on medical implants?

BackgroundBacterial biofilm can occur on all medical implanted devices and lead to infection and/or dysfunction of the device.In this study, artificial biofilm was formed on four different medical implants (silicone, piccline, peripheral venous catheter and endotracheal tube) of interest for our daily clinical and/or research practice. We investigated the best conventional technic to dislodge the biofilm on the implants and quantified the number of bacteria. Staphylococcus epidermidis previously isolated from a breast implant capsular contracture on a patient in the university hospital of Dijon was selected for its ability to produce biofilm on the implants. Different technics (sonication, Digest-EUR®, mechanized bead mill, combination of sonication plus Digest-EUR®) were tested and compared to detach the biofilm before quantifying viable bacteria by colony counting.ResultsFor all treatments, the optical and scanning electron microscope images showed substantial less biofilm biomass remaining on the silicone implant compared to non-treated implant.This study demonstrated that the US procedure was statistically superior to the other physical treatment: beads, Digest-EUR® alone and Digest-EUR® + US (p < 0.001) for the flexible materials (picc-line, PIV, and silicone). The number of bacteria released by the US is significantly higher with a difference of 1 log on each material. The result for a rigid endotracheal tube were different with superiority for the chemical treatment dithiothreitol: Digest-EUR®. Surprisingly the combination of the US plus Digest-EUR® treatment was consistently inferior for the four materials.ConclusionsDepending on the materials used, the biofilm dislodging technique must be adapted. The US procedure was the best technic to dislodge S. epidermidis biofilm on silicone, piccline, peripheral venous catheter but not endotracheal tube. This suggested that scientists should compare themselves different methods before designing a protocol of biofilm study on a given material.