Methicillin-resistant Staphylococcus Aureus Growth Research Articles

Multi-omics revealed antibacterial mechanisms of licochalcone A against MRSA and its antimicrobic potential on pork meat

Licorice flavonoids (LFs) exhibit potent antibacterial activities against Gram-positive bacteria. However, the related mechanism remains unclear. This study aims to illustrate the mechanisms of licochalcone A (LA), a main flavonoid in LFs, against methicillin-resistant Staphylococcus aureus (MRSA). The anti-MRSA effect of LA was comprehensively investigated by a combination of proteomics and metabolomics studies. Meanwhile, LA was loaded in glycyrrhizin (GA) micelles (GA@LA micelles) to improve its water solubility. The results demonstrated that LA could disrupt the arginine metabolism and cause the accumulation of intracellular ROS in MRSA. In addition, LA could inhibit the expression of glucokinase in MRSA, which affect the synthesis of ATP, fatty acids, and peptidoglycan. GA@LA micelles have the latent ability to inhibit the growth of MRSA on fresh pork.

Hyaluronic Acid-Modified Micelles of Azithromycin and Quercetin Against Infections Caused by Methicillin-Resistant Staphylococcus Aureus.

Resistance of intracellular pathogens is a challenge in microbial therapy. Methicillin-resistant Staphylococcus aureus (MRSA), which is able to persist inside the cells of infected tissues, is protected from attack by the immune system and many antimicrobial agents. To overcome these limitations, nano-delivery systems can be used for targeted therapy of intracellular MRSA. Hyaluronic acid-modified azithromycin/quercetin micelles (HA-AZI/Qe-M) were synthesized by thin film hydration. The micelles were characterized by transmission electron microscopy (TEM), dynamic light scattering (DLS) and Fourier transform infrared spectroscopy (FTIR), and the drug loading (DL) and encapsulation efficiency (EE) were detected by high performance liquid chromatography (HPLC). The uptake ability of RAW264.7 cells was investigated, and its distribution in mice was evaluated by in vivo imaging. The inhibitory effect of the micelles against MRSA in vitro and its ability to eliminate intracellular bacteria were evaluated. Bacterial muscle-infected mice were constructed to evaluate the therapeutic effect of the micelles on bacterial infections in vivo and the biocompatibility of the micelles was investigated. HA-AZI/Qe-M had suitable physical and chemical properties and characterization. In vitro antibacterial experiments showed that HA-AZI/Qe-M could effectively inhibit the growth of MRSA, inhibit and eliminate the biofilm formed by MRSA, and have an excellent therapeutic effect on intracellular bacterial infection. The results of RAW264.7 cells uptake and in vivo imaging showed that HA-AZI/Qe-M could increase the cellular uptake, target the infection site, and prolong the treatment time. The results of in vivo antibacterial infection experiments showed that HA-AZI/Qe-M was able to ameliorate the extent of thigh muscle infections in mice and reduce the expression of inflammatory factors. HA-AZI/Qe-M is a novel and effective nano-drug delivery system that can target intracellular bacterial infection, and it is expected to be safely used for the treatment of MRSA infection.

Evaluation of Antibacterial Activity of Fruit Juices on Clinical and non-Clinical Enterotoxigenic, biofilm-producing Methicillin Resistance Staphylococcus aureus

The drug-resistance behavior of Methicillin-resistant Staphylococcus aureus (MRSA) has made it difficult to treat. This study aimed to discover some fruit juice's anti-MRSA potential and the presence of enterotoxin genesin MRSA.MRSA strains were confirmed phenotypically by the disc diffusion method. The well-diffusion method and Minimum inhibitory concentration (MIC)were used to determine the antibacterial effects of fruit juices. Congo red test and ring tests were performed to analyze the biofilm-forming ability. PCR detected staphylococcal enterotoxin genes Sea and Seg. A total of 90strains, of which 26 nonclinical and 64 clinical samples were processed. Ciprofloxacin (CIP) was highly resistant to MRSA in both groups. In clinical isolates, Citrus reticulate has shown the maximum antibacterial activity against 67.1% MRSA, whereas, Punicagranatum was most effective for 46.1% strains isolated from non-clinical sources. Punicagranatum and Citrus reticulate were inhibiting the growth of MRSA at the concentration of >64μl/ml in both groups. Both Congo red and ring tests determined the biofilm-forming ability in 57.60% and 65.30% of non-clinical strains, respectively. Enterotoxin-producing gene Sea was detected in 8% of MRSA in the non-clinical group and only 2% in the clinical group. Seg and the co-existence of both genes were found in the same ratio in both groups. This study showed that fruit juices, especially pomelo, orange, and pomegranate, have high antibacterial activity. Enterotoxin genes play a role in the spread of infections caused by MRSA.

Ginkgo biloba L. exocarp petroleum ether extract inhibits methicillin-resistant Staphylococcus aureus by modulating ion transport, virulence, and biofilm formation in vitro and in vivo

Ethnopharmacological relevanceAs reported in the Ancient Chinese Medicinal Books, Ginkgo biloba L. fruit has been used as a traditional Chinese medicine for the treatment asthma and cough or as a disinfectant. Our previous study demonstrated that G. biloba exocarp extract (GBEE), an extract of a traditional Chinese herb, inhibits the formation of methicillin-resistant Staphylococcus aureus (MRSA) biofilms. However, GBEE is a crude extract that contains many components, and the underlying mechanisms of purified GBEE fractions extracted with solvents of different polarities are unknown. Aim of the studyThis study aimed to investigate the different components in GBEE fractions extracted with solvents of different polarities and their antibacterial effects and mechanisms against MRSA and Staphylococcus haemolyticus biofilms both in vitro and in vivo. MethodsThe components in different fractions were detected by high-performance liquid chromatography–high resolution mass spectrometry (HPLC–HRMS). Microbroth dilution assays and time growth curves were used to determine the antibacterial effects of the fractions on 15 clinical bacterial isolates. Crystal violet staining, scanning electron microscopy (SEM) and transmission electron microscopy (TEM) were utilized to identify the fractions that affected bacterial biofilm formation. The potential MRSA targets of the GBEE fraction obtained with petroleum ether (PE), denoted GBEE-PE, were screened by transcriptome sequencing, and the gene expression profile was verified by quantitative polymerase chain reaction (qPCR). ResultsHPLC–HRMS analysis revealed that the four GBEE fractions (extracted with petroleum ether, ethyl acetate, n-butanol, and water) contained different ginkgo components, and the antibacterial effects decreased as the polarity of the extraction solvent increased. The antibacterial activity of GBEE-PE was greater than that of the GBEE fraction extracted with ethyl acetate (EA). GBEE-PE improved H. illucens survival and reduced MRSA colonization in model mouse organs. Crystal violet staining and SEM and TEM analyses revealed that GBEE-PE inhibited MRSA and S. haemolyticus biofilm formation. Transcriptional analysis revealed that GBEE-PE inhibits MRSA biofilms by altering ion transport, cell wall metabolism and virulence-related gene expression. In addition, the LO2 cell viability and H. illucens toxicity assay data showed that GBEE-PE at 20 mg/kg was nontoxic. ConclusionThe GBEE fractions contained different components, and their antibacterial effects decreased with increases in the polarity of the extraction solvent. GBEE-PE limited MRSA growth and biofilm formation by affecting ion transport, cell wall synthesis, and virulence-related pathways. This research provides a more detailed overview of the mechanism by which GBEE-PE inhibits MRSA both in vitro and in vivo and suggests that GBEE-PE is a new prospective antimicrobial with the potential to be used in MRSA therapeutics in the future.

Antibacterial and antibiofilm activities of protocatechualdehyde and its synergy with ampicillin against methicillin-resistant Staphylococcus aureus.

Protocatechualdehyde (PA) is a phenolic acid present in many plants and has many biological activities. Herein, the antagonistic effects and the action mechanism of PA against methicillin-resistant Staphylococcus aureus (MRSA) were studied. The results showed that PA had both significant antibacterial and anti-biofilm activities against MRSA. Additionally, PA had synergy with ampicillin against MRSA. It was elucidated that PA was prominent in destroying cell membranes, increasing cell membrane permeability and intracellular ROS production, thus leading to bacterial cell damage. Transcriptome analysis showed that PA disrupts many physiological pathways, including increasing cell membrane permeability, inhibiting biofilm formation, decreasing resistance to antimicrobial agents, and impairing DNA replication. Finally, the antimicrobial preservation test showed that PA could inhibit the growth of MRSA and prevent the corruption of beef. In summary, PA is an effective natural antibacterial substance and has a good application potential in food preservation, even in tackling antibiotic resistance problems.

Endophytic fungi of Tradescantia pallida mediated targeting of Multi-Drug resistant human pathogens

Antimicrobial resistance (AMR) has emerged as one of the most serious worldwide public health issues of the twenty-first century. The expeditious rise of AMR has urged the development of new, natural effective therapeutic strategies against drug-resistant pathogens. Endophytic fungi, which inhabit distinctive environments like endosymbiotic relationships with plants, are gaining interest as alternative reservoirs for novel compounds that exhibit a broad range of chemical diversity and unique modes of action by releasing a variety of secondary metabolites with antimicrobial properties. The objective of the current research was to isolate and identify endophytic fungal species from leaves of Tradescantia pallida and to investigate their antagonistic effects on Multi-Drug-Resistant human pathogens. Endophytic fungus TPL11 and TPL14 showed maximum inhibition in agar plug and agar well diffusion assay. The ethyl acetate crude extract effectively suppressed growth of MRSA (Methicillin-resistant Staphylococcus aureus) ATCC 43300,700699 strains and VRE (Vancomycin-resistant Enterococcus) with the Inhibition zone of 22 ± 0.05, 23 ± 0.11 and 24 ± 0.11 mm respectively with minimum inhibitory concentration (MIC) of 3.125 µg/mL. Whereas TPL11 fungus revealed antibiosis of 22 ± 0.05 and 21 ± 0.15 mm against MRSA(ATCC 43300,700699) and 24 ± 0.05 mm for VRE with MIC of 6.25,3.125 and 1.56 μg/mL respectively. The MIC (Minimum inhibitory concentration) index further confirmed that both the extracts were bacteriostatic against MRSA and bactericidal against VRE. The isolates TPL11 and TPL14 were identified as Fusarium oxysporum and Nigrospora sphaerica by 18S rRNA internal transcribed spacer (ITS) sequencing. To our insight, it is the first report to reveal the presence of F.oxysporum and N.sphaerica in T.pallida and their antibacterial activity.

Design and synthesis of vancomycin-functionalized ZnFe2 O4 nanoparticles as an effective antibacterial agent against methicillin-resistant Staphylococcus aureus.

The emergence of antibiotic-resistant bacterial infections is a principal threat to global health. Functionalization of nanomaterial with antibiotics is known as a useful method for increasing the effectiveness of existing antibiotics. In this study, vancomycin-functionalized ZnFe2 O4 nanocomposite (ZnFe2 O4 @Cell@APTES@Van) was synthesized, and its functional groups and particle size were characterized using Fourier-transform infrared spectroscopy, thermogravimetric analysis, dynamic light scattering, scanning electron microscope, and transmission electron microscopy. The antibacteria activity of the synthesized nanocomposite was evaluated using minimum inhibitory concentration and minimum bactericidal concentration against Escherichia coli, Staphylococcus aureus, and methicillin-resistant Staphylococcus aureus (MRSA). Cytotoxicity assay was done by 2,3-bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilidemethod. Characterization analyses of synthesized nanocomposite confirmed the binding of vancomysin on the surface of ZnFe2 O4 @Cell@APTES. Nanocomposite exhibited an aggregated semi-spherical structure with hydrodynamic radii of ∼382 nm. In vitro antibacterial activity test showed that vancomycin and vancomycin functionalized ZnFe2 O4 have no antibacterial effect against E. coli. S. aureus was sensitive to vancomycin and ZnFe2 O4 @Cell@APTES@Van NPs and ZnFe2 O4 NPs did not improve vancomycin antibacterial activity against these bacteria. MRSA is resistant to vancomycin while ZnFe2 O4 @Cell@APTES@Van NPs was efficient in inhibiting MRSA growth. In summary, this study showed that attachment of vancomycin to ZnFe2 O4 NPs was increased its antibacterial activity against MRSA.

COMBATTING METHICILLIN-RESISTANT STAPHYLOCOCCUS AUREUS (MRSA) IN THE FOOD INDUSTRY BY HARNESSING THE POWER OF NATURE: A SYSTEMATIC REVIEW

Antibiotic resistance is a critical global health concern, with Methicillin-resistant Staphylococcus aureus (MRSA) posing a significant challenge due to its resistance to commonly used antibiotics. Recent research has revealed the potential of natural compounds and microorganisms in combatting MRSA and other antibiotic-resistant bacteria. In this systematic review, we studied the effect of essential oils, bacteriophages, bacteriocins, and probiotics on S. aureus, including MRSA in particular, in the food industry. Essential oils (EOs) have gained significant attention because of their antimicrobial properties, inhibiting MRSA growth by damaging bacterial cells and inhibiting essential enzymes and compounds. Cinnamon oil liposomes caused the most significant decrease in MRSA populations among our reviewed essential oils. Bacteriophages can lyse the bacterial host. They encode peptidoglycan hydrolases called endolysins that target the bacterial cell wall. In our study, S. aureus phage (containing CHAPLysGH15 and LysGH15), and phage SA11 endolysin (LysSA11) were the most effective against S. aureus. Bacteriocins, antimicrobial peptides produced by bacteria, also show potential in combatting MRSA, mainly by generating organic acids that interfere with bacterial metabolism. According to our review, the most effective bacteriocins against S. aureus were Enterocin AS-48 with phenolic compounds or with 2NPOH, Bacteriocin isolated from Lactobacillus pentosus - Pentocin JL-1, and bacteriocin produced by S. pasteuri RSP-1, respectively. Probiotics can compete with pathogens by producing antimicrobial compounds that disrupt MRSA cell production and ultimately lead to bacterial death. In our review, the most effective probiotics were Streptomyces griseus, Pediococcus acidilactici strains A11 and C12, Lactococcus lactis, and Lactobionic acid respectively. A multi-hurdle approach combining these natural agents has shown promising results in targeting and eliminating MRSA cells. By harnessing the power of nature, we can potentially overcome the challenges posed by MRSA and other antibiotic-resistant bacteria.

Growth of methicillin-resistant Staphylococcus aureus during raw milk soft cheese-production and the inhibitory effect of starter cultures

The consumption of raw milk or raw milk products might be a potential risk factor for the transmission of methicillin-resistant Staphylococcus aureus (MRSA). Therefore, we studied MRSA growth during raw milk soft cheese-production. Furthermore, we investigated the inhibitory effect of four starter cultures (Lactococcus lactis, Lacticaseibacillus rhamnosus, Lactiplantibacillus plantarum, Lactobacillus helveticus) on the growth of MRSA in a spot-agar-assay and in raw milk co-culture following a cheesemaking temperature profile. During the initial phases of raw milk cheese-production, MRSA counts increased by 2 log units. In the ripening phase, MRSA counts only dropped slightly and remained high up to the end of the storage. Comparable MRSA counts were found in the rind and core and strain-specific differences in survival were observed. In the spot-agar-assay, all four starter cultures showed strong or intermediate inhibition of MRSA growth. In contrast, in raw milk, only Lactococcus lactis strongly inhibited MRSA, whereas all other starter cultures only had minor inhibitory effects on MRSA growth. Our results indicate that MRSA follow a similar growth pattern as described for other S. aureus during raw milk soft cheese-production and illustrate the potential use of appropriate starter cultures to inhibit MRSA growth during the production of raw milk cheese.

Bacterial pneumonia-induced shedding of epithelial heparan sulfate inhibits the bactericidal activity of cathelicidin in a murine model.

Bacterial pneumonia is a common clinical syndrome leading to significant morbidity and mortality worldwide. In the current study, we investigate a novel, multidirectional relationship between the pulmonary epithelial glycocalyx and antimicrobial peptides in the setting of methicillin-resistant Staphylococcus aureus (MRSA) pneumonia. Using an in vivo pneumonia model, we demonstrate that highly sulfated heparan sulfate (HS) oligosaccharides are shed into the airspaces in response to MRSA pneumonia. In vitro, these HS oligosaccharides do not directly alter MRSA growth or gene transcription. However, in the presence of an antimicrobial peptide (cathelicidin), increasing concentrations of HS inhibit the bactericidal activity of cathelicidin against MRSA as well as other nosocomial pneumonia pathogens (Klebsiella pneumoniae and Pseudomonas aeruginosa) in a dose-dependent manner. Surface plasmon resonance shows avid binding between HS and cathelicidin with a dissociation constant of 0.13 μM. These findings highlight a complex relationship in which shedding of airspace HS may hamper host defenses against nosocomial infection via neutralization of antimicrobial peptides. These findings may inform future investigation into novel therapeutic targets designed to restore local innate immune function in patients suffering from primary bacterial pneumonia.NEW & NOTEWORTHY Primary Staphylococcus aureus pneumonia causes pulmonary epithelial heparan sulfate (HS) shedding into the airspace. These highly sulfated HS fragments do not alter bacterial growth or transcription, but directly bind with host antimicrobial peptides and inhibit the bactericidal activity of these cationic polypeptides. These findings highlight a complex local interaction between the pulmonary epithelial glycocalyx and antimicrobial peptides in the setting of bacterial pneumonia.

Antibacterial and antibiofilm activity of Abroma augusta stabilized silver (Ag) nanoparticles against drug-resistant clinical pathogens.

Infectious diseases remain among the most pressing concerns for human health. This issue has grown even more complex with the emergence of multidrug-resistant (MDR) bacteria. To address bacterial infections, nanoparticles have emerged as a promising avenue, offering the potential to target bacteria at multiple levels and effectively eliminate them. In this study, silver nanoparticles (AA-AgNPs) were synthesized using the leaf extract of a medicinal plant, Abroma augusta. The synthesis method is straightforward, safe, cost-effective, and environment friendly, utilizing the leaf extract of this Ayurvedic herb. The UV-vis absorbance peak at 424nm indicated the formation of AA-AgNPs, with the involvement of numerous functional groups in the synthesis and stabilization of the particles. AA-AgNPs exhibited robust antibacterial and antibiofilm activities against methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin-resistant Enterococci (VRE). The MIC values of AA-AgNPs ranged from 8 to 32μg/mL. Electron microscopic examination of the interaction of AA-AgNPs with the test bacterial pathogens showed a deleterious impact on bacterial morphology, resulting from membrane rupture and leakage of intracellular components. AA-AgNPs also demonstrated a dose-dependent effect in curtailing biofilm formation below inhibitory doses. Overall, this study highlights the potential of AA-AgNPs in the successful inhibition of both the growth and biofilms of MRSA and VRE bacteria. Following studies on toxicity and dose optimization, such AgNPs could be developed into effective medical remedies against infections.

In Vitro Synergistic Inhibitory Effects of Plant Extract Combinations on Bacterial Growth of Methicillin-Resistant Staphylococcus aureus.

Methicillin-resistant Staphylococcus aureus (MRSA) is one of the most common pathogens of healthcare-associated infections. Medicinal plants have long been used in the traditional treatment of diseases or syndromes worldwide. Combined use of plant extracts could improve the effectiveness of pharmacological action by obtaining synergism, acting on multiple targets simultaneously, reducing the doses of individual components, and minimizing side effects. We aimed to investigate the synergistic inhibitory effects of selected medicinal plants (Caesalpinia sappan L. (CS), Glycyrrhiza uralensis Fisch. (GU), Sanguisorba officinalis L. (SO), and Uncaria gambir Roxb. (UG)) on the bacterial growth of MRSA and its clinical isolates. SO and UG extracts generated the best synergistic interaction as adjudged by checkerboard synergy assays. MICs of the individual extracts decreased 4-fold from 250 to 62.5 μg/mL, respectively. The SO + UG combination was further evaluated for its effects on bacterial growth inhibition, minimum bactericidal/inhibitory concentration (MBC/MIC) ratio, and time-kill kinetics. The results indicate that the SO + UG combination synergistically inhibited the bacterial growth of MRSA strains with bactericidal effects. SO + UG combination also exhibited more potent effects against clinical isolates. In multistep resistance selection experiments, both standard and isolates of MRSA showed no resistance to the SO + UG combination even after repeated exposure over fourteen passages. Our data suggest that using plant extract combinations could be a potential strategy to treat MRSA infections.

Longitudinal intravital imaging of the bone marrowfor analysis of the race for the surface in a murine osteomyelitis model.

Critical knowledge gaps of orthopedic infections pertain to bacterial colonization. The established dogma termed the Race for the Surface posits that contaminating bacteria compete with host cells for the implant post-op, which remains unproven without real-time in vivo evidence. Thus, we modified the murine longitudinal intravital imaging of the bone marrow (LIMB) system to allow real-time quantification of green fluorescent protein(GFP+) host cells and enhanced cyan fluorescent protein(ECFP+) or red fluorescent protein(RFP+) methicillin-resistant Staphylococcus aureus (MRSA) proximal to a transfemoral implant. Following inoculation with ~105 CFU, an L-shaped metal implant was press-fit through the lateral cortex at a 90° angle ~0.150 mm below a gradient refractive index (GRIN) lens. We empirically derived a volume of interest (VOI) = 0.0161 ± 0.000675 mm3 during each imaging session by aggregating the Z-stacks between the first (superior) and last (inferior) in-focus LIMB slice. LIMB postimplantation revealed very limited bacteria detection at 1 h, but by 3 h,56.8% of the implant surface was covered by ECFP+ bacteria, and the rest were covered by GFP+ host cells. 3D volumetric rendering of the GFP+ and ECFP+ or RFP+ voxels demonstrated exponential MRSA growth between 3 and 6 hin the Z-plane, which was validated with cross-sectional ex vivo bacterial burden analyses demonstrating significant growth by ~2 × 104 CFU/hon the implant from 2 to 12 hpost-op (p < 0.05; r2 > 0.98). Collectively, these results show the competition at the surface is completed by 3 hin this model and demonstrate the potential of LIMB to elucidate mechanisms of bacterial colonization, the host immune response, and the efficacy of antimicrobials.

In Situ Photoacoustic Visualization of Pneumonia Induced by MRSA and Specific Identifying Tumor-Homing Bacteria.

Optical imaging holds great promise for monitoring bacterial infectious processes and drug resistance with high temporal-spatial resolution. Currently, the diagnosis of deep-seated bacterial infections in vivo with fluorescence imaging, including near-infrared (NIR) fluorescence imaging technology, remains a significant challenge due to its limited tissue penetration depth. In this study, we developed a highly specific targeting probe, Cy7-Neo-NO2, by conjugating a bacterial 16S rRNA-targeted moiety, neomycin, with a bacterial nitroreductase (NTR)-activated NIR photoacoustic (PA) scaffold using our previously developed caged photoinduced electron transfer (a-PeT) approach. This conjugation effectively resolved probe aggregation issues in physiological conditions and substantially enhanced its reactivity toward bacterial NTR. Notably, Cy7-Neo-NO2 enabled the first in situ photoacoustic imaging of pneumonia induced by methicillin-resistant Staphylococcus aureus (MRSA), as well as the detection of bacteria within tumors. Furthermore, upon NIR irradiation, Cy7-Neo-NO2 successfully inhibited MRSA growth through a synergistic effect combining photothermal therapy and photodynamic therapy. Our results provided an effective tool for obtaining exceptional PA agents for accurate diagnosis, therapeutic evaluation of deep-seated bacterial infections in vivo, and intratumoral bacteria-specific recognition.

Liposome-loaded polymeric microneedles for enhanced skin deposition of rifampicin

Methicillin-resistant Staphylococcus aureus (MRSA) is a prevailing bacterial pathogen linked to superficial skin and soft tissue infections (SSTIs). Rifampicin (RIF), a potent antibiotic against systemic and localised staphylococcal infections, faces limitations due to its low solubility. This constraint hampers its therapeutic potential for MRSA-induced SSTIs. To address this, an advanced liposomal system was designed for efficient dermal RIF delivery. Rifampicin-loaded liposomes (LipoRIF) were embedded within polymeric dissolving microneedles (DMNs) to enable targeted intradermal drug delivery. A robust Design of Experiment (DoE) methodology guided the systematic preparation and optimisation of LipoRIF formulations. The optimal LipoRIF formulation integrated within polymeric DMNs. These LipoRIF-DMNs exhibited favourable mechanical properties and effective skin insertion characteristics. Notably, in vitro assays on skin deposition unveiled a transformative result – the DMN platform significantly enhanced LipoRIF deposition within the skin, surpassing LipoRIF dispersion alone. Moreover, LipoRIF-DMNs displayed minimal cytotoxicity toward cells. Encouragingly, rigorous in vitro antimicrobial evaluations demonstrated LipoRIF-DMNs' capacity to inhibit MRSA growth compared to the control group. LipoRIF-DMNs propose a potentially enhanced, minimally invasive approach to effectively manage SSTIs and superficial skin ailments stemming from MRSA infections.

In Vitro and In Vivo Antimicrobial Activity of an Oxidative Stress-Mediated Bicyclic Menaquinone Biosynthesis Inhibitor against MRSA.

Menaquinone (MK) is an essential component in the oxidative phosphorylation pathway of Gram-positive bacteria. Drugs targeting enzymes involved in MK biosynthesis can prevent electron transfer, which leads to ATP starvation and thereby death of microorganisms. Previously, we reported a series of MenA inhibitors and demonstrated their antimicrobial activity against Gram-positive bacteria, including Methicillin-resistant Staphylococcus aureus (MRSA) and mycobacteria. These inhibitors were developed by mimicking demethylmenaquinone, a product of MenA enzymatic reaction in MK biosynthesis. In this study, compound NM4, MK biosynthesis inhibitor, inhibited the formation of MRSA biofilm and it was screened against 1952 transposon mutants to elucidate mechanisms of action; however, no resistant mutants were found. Also, compound NM4 induced the production of reactive oxygen species (ROS) by blocking electron transfer in the oxidative phosphorylation pathway as observed by MRSA growth recovery using various ROS scavengers. An oxygen consumption assay also showed that NM4 blocks the oxygen consumption by MRSA, but the addition of menaquinone (MK) restores growth of MRSA. The NM4-treated MRSA induced the expression of catalase by more than 25%, as quantified by the native gel. A pulmonary murine model exhibited that NM4 significantly reduced bacterial lung load in mice without toxicity. An NM4-resistant USA300 strain was developed to attempt to identify the targets participating in the mechanism of resistance. Our results support that respiration and oxidative phosphorylation are potential targets for developing antimicrobial agents against MRSA. Altogether, our findings suggest the potential use of MK biosynthesis inhibitors as an effective antimicrobial agent against MRSA.

Inhibitory Effect of Non-Saccharomyces Starmerella bacillaris CC-PT4 Isolated from Grape on MRSA Growth and Biofilm.

Methicillin-resistant Staphylococcus aureus (MRSA) is a notorious pathogen with biofilm-forming and drug-resistant properties that make it difficult to eradicate. In this study, the inhibition of MRSA (ATCC 43300) by Starmerella bacillaris CC-PT4 (CGMCC No. 23573) was evaluated. The results showed that the inhibition of MRSA growth and biofilm was caused by S. bacillaris CC-PT4 cell-free supernatant (CFS). The CFS of S. bacillaris CC PT4 at different times can effectively inhibit the formation of MRSA biofilm, remove the preformed biofilm, and down-regulate the related genes that promote the formation of biofilm. Afterwards, untargeted metabolomics was performed to analyze the CFS of S. bacillaris CC-PT4. Several molecules with antibacterial and inhibitory biofilm effects from the CFS were found, one of which, 2-amino-1-phenylethanol (APE), has not been reported to have antiMRSA ability before. In this study, molecular docking analysis and in vitro experiments were used to verify the function of APE to inhibit MRSA. These results indicate that S. bacillaris CC-PT4 CFS can effectively inhibit MRSA which has potential application value in controlling MRSA.

Mathematical modeling of the combined action of Origanum grosii essential oil and sodium chloride on the growth of methicillin-resistant Staphylococcus aureus

Methicillin-resistant Staphylococcus aureus (MRSA) is one of the emerging opportunistic pathogens that can cause food poisoning and is responsible for several infectious diseases, including hospital-acquired infections. The main objective of this work was to assay the antibacterial effect of Origanum grosii essential oil (OGEO) and NaCl, alone and in combination against the MRSA growth. For this purpose, response surface methodology (RSM) was adopted to generate a mathematical model representing this effect. O. grosii was harvested from the north of Morocco. GC-FID and GC/MS techniques were used in order to determine the chemical profile of OGEO. The main compounds of OGEO were carvacrol (49.29%), thymol (13.12%), p-cymene (12.68%), and γ-terpinene (8.78%). The method of micro-dilution was used in order to determine the minimal inhibitory concentration (MIC) and the minimal bactericidal concentration (MBC). The MIC value of OGEO was 0.5% (v/v), while the MBC value was equal to 1%(v/v). The OGEO showed a strong bactericidal effect against MRSA. RSM applied at two different incubation times, 6 h and 11 h, and at the total time average value (TAV), allowed to create a mathematical model representing the kinetics of MRSA growth depending to the concentrations of OGEO and salt. The interaction between these two variables was significant. The quadratic polynomial regression was sufficient to adequately represents the observed results with a high coefficient of determination R2 ranging from 95.8 to 96.4%. A synergy at first order was observed between the two factors, but it was not present at all time points; a slight antagonistic effect was observed at time t = 6 h. These results show that RSM provided a useful and accurate mathematical method by combining low sub-inhibitory concentrations of OGEO with NaCl that can be used as a natural product against MRSA infections or as an additive in the food system.

Tannin extracted from Penthorum chinense Pursh, a potential drug with antimicrobial and antibiofilm effects against methicillin-sensitive Staphylococcus aureus and methicillin-resistant Staphylococcus aureus.

Staphylococcus aureus is an opportunistic pathogen. Due to the widespread use and abuse of antibiotics, various drug-resistant strains of S. aureus have emerged, with methicillin-resistant Staphylococcus aureus (MRSA) being the most prevalent. Bacterial biofilm is a significant contributor to bacterial infection and drug resistance. Consequently, bacterial biofilm formation has emerged as a therapeutic strategy. In this study, the chemical constituents, antimicrobial and antibiofilm properties of tannins isolated from Penthorum chinense Pursh (TPCP) were investigated. In vitro, TPCP exhibited antimicrobial properties. The minimum inhibitory concentrations (MIC) and minimum bactericidal concentrations (MBC) for methicillin-sensitive Staphylococcus aureus (MSSA) and MRSA were 156.25 and 312.5 μg/mL, and 312.5 and 625 μg/mL, respectively. According to the growth curves, TPCP significantly inhibited the growth of MSSA and MRSA. The results of the crystal violet biofilm assay in conjunction with confocal laser scanning and scanning electron microscopy demonstrated that TPCP destroyed preformed MSSA and MRSA biofilms. TPCP significantly decreased the secretion of exopolysaccharides and extracellular DNA. Subsequently, the mechanism was investigated using RT-PCR. Examining the expression of icaA, cidA, sigB, agrA, and sarA genes in MRSA, we discovered that TPCP inhibited biofilm formation by affecting the quorum-sensing system in bacteria. Our study demonstrates that TPCP exerts antibacterial effects by disrupting the formation of bacterial biofilms, suggesting that TPCP has clinical potential as a novel antibacterial agent for the prevention and treatment of MSSA and MRSA infections.

Harnessing the power of PLA-PEG nanoparticles for Linezolid delivery against methicillin-resistant Staphylococcus aureus

This study deals with the development of novel poly(lactic acid)-poly(ethylene glycol) nanoparticles (PLA-PEG NPs) for the efficient and prolonged delivery of Linezolid (LNZ), a synthetic antibacterial agent used against methicillin-resistant Staphylococcus aureus (MRSA). A two-step synthetic strategy based on carbodiimide coupling and copper-catalyzed azide-alkyne cycloaddition was first exploited for the conjugation of PLA with PEG. The encapsulation of LNZ into medium-molecular-weight PLA-PEG NPs was carried out by different methods including nanoprecipitation and dialysis. The optimal PLA-PEG@LNZ nanoformulation resulted in 3.5% LNZ payload (15% encapsulation efficiency, with a 10:3 polymer to drug mass ratio) and sustained release kinetics with 65% of entrapped antibiotic released within 80 h. Moreover, the zeta potential values (from −31 to −39 mV) indicated a good stability without agglomeration even after freeze-drying and lyophilization. The PLA-PEG@LNZ NPs exerted antimicrobial activity against a panel of Gram-positive bacteria responsible for human infections, such as Staphylococcus aureus including MRSA, Staphylococcus epidermidis, Staphylococcus lugdunensis and vancomycin-resistant Enterococcus faecium (VREfm). Moreover, PLA-PEG@LNZ NPs showed inhibitory activity on both planktonic growth and preformed biofilm of MRSA. The antibacterial activity of LNZ incorporated in polymeric NPs was well preserved and the nanosystem served as an antibiotic enhancer with a potential role in MRSA-associated infections management.