Viability Of Methicillin-resistant Staphylococcus Aureus Research Articles

Flotillin-mediated stabilization of unfolded proteins in bacterial membrane microdomains

The function of many bacterial processes depends on the formation of functional membrane microdomains (FMMs), which resemble the lipid rafts of eukaryotic cells. However, the mechanism and the biological function of these membrane microdomains remain unclear. Here, we show that FMMs in the pathogen methicillin-resistant Staphylococcus aureus (MRSA) are dedicated to confining and stabilizing proteins unfolded due to cellular stress. The FMM scaffold protein flotillin forms a clamp-shaped oligomer that holds unfolded proteins, stabilizing them and favoring their correct folding. This process does not impose a direct energy cost on the cell and is crucial to survival of ATP-depleted bacteria, and thus to pathogenesis. Consequently, FMM disassembling causes the accumulation of unfolded proteins, which compromise MRSA viability during infection and cause penicillin re-sensitization due to PBP2a unfolding. Thus, our results indicate that FMMs mediate ATP-independent stabilization of unfolded proteins, which is essential for bacterial viability during infection.

Inhibition mechanisms of secretome proteins from Paenibacillus polymyxa Kp10 and Lactococcus lactis Gh1 against methicillin-resistant Staphylococcus aureus and vancomycin-resistant Enterococcus

Objective: To determine the inhibition mechanisms of secretome protein extracted from Paenibacillus polymyxa Kp10 (Kp10) and Lactococcus lactis Gh1 (Gh1) against methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin-resistant Enterococcus (VRE).Methods: The sensitivity and viability of MRSA and VRE treated with secretome proteins of Kp10 and Gh1 were determined using minimal inhibitory concentration, minimum bactericidal concentration, and time-to-kill assays. The morphological changes were observed using scanning electron microscopy and transmission electron microscopy. To elucidate the antimicrobial mechanism of secretome protein of Kp10 and Gh1 against MRSA and VRE, 2D gel proteomic analysis using liquid chromatography-mass spectrometry was run by comparing upregulated and downregulated proteins, and the proton motive force study including the efflux of ATP, pH gradient, and the membrane potential study were conducted.Results: MRSA and VRE were sensitive to Kp10 and Gh1 secretome protein extracts and displayed apparent morphological and internal composition changes. Several proteins associated with cellular component functions were either downregulated or upregulated in treated MRSA and VRE by changing the membrane potential gradient.Conclusions: Kp10 and Gh1 secretome proteins reduce the growth of VRE and MRSA by damaging the cell membrane. Cell division, cell wall biosynthesis, and protein synthesis are involved in the inhibition mechanism.

Inhibition of Bacterial Efflux Pumps by Crude Extracts and Essential Oil from Myristica fragrans Houtt. (Nutmeg) Seeds against Methicillin-Resistant Staphylococcus aureus.

Myristicafragrans Houtt. (Nutmeg) is a widely known folk medicine across several parts of Asia, particularly used in antimicrobial treatment. Bacterial resistance involves the expression of efflux pump systems (chromosomal norA and mepA) in methicillin-resistant Staphylococcus aureus (MRSA). Crude extract (CE) and essential oil (EO) obtained from nutmeg were applied as efflux pump inhibitors (EPIs), thereby enhancing the antimicrobial activity of the drugs they were used in. The major substances in CE and EO, which function as EPIs, in a descending order of % peak area include elemicin, myristicin, methoxyeugenol, myristicin, and asarone. Here, we investigated whether the low amount of CE and EO used as EPIs was sufficient to sensitize MRSA killing using the antibiotic ciprofloxacin, which acts as an efflux system. Interestingly, synergy between ciprofloxacin and CE or EO revealed the most significant viability of MRSA, depending on norA and mepA, the latter being responsible for EPI function of EO. Therefore, CE and EO obtained from nutmeg can act as EPIs in combination with substances that act as efflux systems, thereby ensuring that the MRSA strain is susceptible to antibiotic treatment.

Polymeric antibiofilm coating comprising synergistic combination of citral and thymol prevents methicillin-resistant Staphylococcus aureus biofilm formation on titanium

Biomaterial associated microbial infections are complicated and mostly lead to revision surgery or removal which are painful to the patients and quite expensive. These infections are difficult to treat with antibiotics as it is often related to biofilm formation. Methicillin resistant Staphylococcus aureus (MRSA) is the leading pathogen in biomaterial associated infections and well known to form biofilm on foreign materials. To reduce the risk of biomaterial associated infections, recent treatment strategies focus on modification of the implant surface to prevent the adhesion of bacteria. Antibiofilm coating is the effective approach than coating with antimicrobials as antibiofilm agents will not create selective pressure thereby excludes possibility of drug resistance. The current study identified and validated the synergistic antibiofilm activity of citral (CIT) and thymol (THY) by crystal violet quantification and microscopic analysis without alteration in growth and metabolic viability of MRSA. Polymeric antibiofilm coating with CIT + THY as active ingredients was formulated and coated on titanium surface by the process of spin coating. Fourier-transform infrared spectroscopy (FTIR) analysis confirmed the effective blending of polymeric formulation and the presence of CIT and THY. Atomic force microscopy (AFM) images revealed the homogenous coating and reduced surface roughness and thickness of the coating was measured by surface profilometer. Antibiofilm coating released CIT and THY in a sustained manner for 60 days. Antibiofilm coating effectively inhibited MRSA adherence in vitro and antibiofilm activity of coating was not affected by plasma conditioning. In addition, antibiofilm coating was non-hemolytic and non-toxic to PBMC. Thus, the current study demonstrated the effectual strategy to prevent biomaterial associated infections and proposes the prospective role of antibiofilm coating in clinical applications.

Indoor Finish Material Influence on Contamination, Transmission, and Eradication of Methicillin-Resistant Staphylococcus aureus (MRSA).

Objective:The purpose of this study was to evaluate environmental surface materials used in healthcare environments for material composition, methicillin-resistant Staphylococcus aureus (MRSA) viability, and a comparison of two disinfectants, a bleach germicidal cleaner and Decon7, a novel disinfectant.Background:Contaminated environmental surfaces have been associated with outbreaks of healthcare-associated illness (HAIs). One in every 20 patients in U.S. acute care hospitals acquire a healthcare-associated illness, leading to consequences such as elevated morbidity, mortality, and a decrease in quality of life. In the patient environment, MRSA can remain viable from hours to up to 14 days.Methods:Environmental surface materials were evaluated as new and worn. Material composition and properties were assessed to evaluate surface integrity and the influence on the disinfection of MRSA. Inoculated materials were used to assess MRSA viability over time and the efficacy of a manufacturer’s recommended cleaning and disinfection product compared to a novel disinfectant.Results:Environmental surface materials respond differently in appearance and roughness, when mechanically worn. When measuring MRSA survival, at 24 hr, MRSA colony forming unit (CFU) counts were reduced on the copper sheet surface and solid surface with cupric oxide. By 72 hr, all MRSA counts were zero. Bleach and the novel disinfectant were equally effective at disinfecting MRSA from all surface types.Conclusions:This study highlights a gap in knowledge about the impact of type and wear of environmental surface materials used in healthcare environments on contamination with epidemiologically important organisms. In conclusion, environmental surface material wear, properties, and cleaning and disinfection efficacy are important factors to consider when addressing HAIs.

Predictors of Surgical Site Infections Among Patients With Diabetes Mellitus Post Coronary Artery Bypass Graft Surgery: A Quasi-experimental Study

Biofilm in chronic wounds impedes the wound healing process. Each biofilm has differing characteristics requiring a multifaceted approach for removal while maintaining a surrounding environment conducive to wound healing. In this study, 3 of the components in a wound cleanser are tested to determine synergy in eradicating biofilms of methicillin-resistant Staphylococcus aureus (MRSA) and Pseudomonas aeruginosa in vitro. The 3 components assessed for synergy were ethylenediamine tetraacetic acid sodium salts (EDTA), vicinal diols (VD; ethylhexylglycerin and octane-1,2-diol), and polyhexamethylene biguanide (PHMB). Each component was assessed individually and in combination while dissolved in a base solution. The Calgary assay method was used for biofilm growth and treatment. Kull Equation analysis for synergy was conducted using viable count results. Synergy is defined as the interaction of components to produce a combined effect greater than the sum of their separate effects. The base solution containing all 3 components (EDTA, VD, and PHMB) reduced biofilm viability by more than 5 logs, demonstrating statistically significant synergy. The 3 components tested individually in the base solution resulted in the following: EDTA did not reduce bacteria viability; VD reduced viability by about 1 log; and PHMB reduced P aeruginosa viability by about 2.5 logs and MRSA viability by about 4 logs. Of importance, the MRSA biofilm failed to regrow in the recovery plates after combined treatment, indicating complete elimination of the biofilm bacteria. The experimental and calculated results indicate the 3 components (VD, EDTA, and PHMB) when used together act synergistically to eradicate MRSA and P aeruginosa biofilms in vitro.

PH-Responsive Micelles From an Oleic Acid Tail and Propionic Acid Heads Dendritic Amphiphile for the Delivery of Antibiotics

The aim of this study was to synthesize a novel biocompatible pH-responsive oleic acid-based dendritic lipid amphiphile (OLA-SPDA) which self-assembled into stable micelles (OLA-SPDA -micelles) with a relatively low critical micelle concentration (CMC) of 5.6 × 10−6 M. The formulated micelles had particle size, polydispersity index (PDI) and zeta potential (ZP) of 84.16 ± 0.184 nm, 0.199 ± 0.011 and −42.6 ± 1.98 mV, respectively, at pH 7.4. The vancomycin (VCM) encapsulation efficiency was 78.80 ± 3.26%. The micelles demonstrated pH-responsiveness with an increase in particle size to 141.1 ± 0.0707 nm and a much faster release profile at pH 6.0, as compared to pH 7.4. The minimum inhibitory concentration (MIC) of VCM-OLA-SPDA-micelle against methicillin-resistant staphylococcus aureus (MRSA) was 8-fold lower compared to bare VCM, and the formulation had a 4-fold lower MIC at pH 6.0 when compared to the formulation's MIC at pH 7.4. MRSA viability assay showed the micelles had a percentage killing of 93.39% when compared bare-VCM (58.21%) at the same MIC (0.98 μg/mL). In vivo mice (BALB/c) skin infection models showed an 8-fold reduction in MRSA burden after treatment with VCM-OLA-SPDA-micelles when compared with bare VCM. The above results suggest that pH-responsive VCM-OLA-SPDA-micelles has the potential to be an effective carrier to enhance therapeutic outcomes against infections characterised by low pH.

The Fate of Methicillin-Resistant Staphylococcus aureus in a Synthetic Turf System.

Methicillin-resistant Staphylococcus aureus (MRSA) is responsible for one-third of infectious outbreaks reported among competitive athletes at high school and collegiate levels of competition, while the frequency of infections among National Football League players is nearly 400-fold higher than in the general population. The increased prevalence of MRSA infections among such athletes necessitates the study of football-specific environments as MRSA reservoirs. The recovery and viability of MRSA will differ between turf infill and turf fibers. Descriptive laboratory study. Level 4. Each of 6 infill types and 3 turf fiber types were inoculated with MRSA, sacrificed over time, and enumerated after bacteria extraction and dilution plating to determine the incubation time at which 50% of inoculated MRSA are recoverable and viable (here termed A50). The role of infill toxicity on MRSA survival was assessed using a dialysis assay. On average, MRSA was available for up to 96 hours on infill (average A50: 13 hours) and 24 hours on turf fibers (A50: 4 hours) (P < 0.001). The A50 for each infill type was also variable among infill groups (P < 0.001), averaging 2 hours (ethylene propylene diene monomer [EPDM] rubber), 7 hours (cork-based material), 9 hours (polymer-coated materials), 12 hours (crumb rubber), 13 hours (thermoplastic elastomer [TPE] rubber), and 27 hours (sands). MRSA remained available on slit-film and nylon fiber types for 12 hours postdeposition and for 24 hours on monofilament fibers. Toxicity assays showed that over 90% of MRSA cells remained viable after 6 hours of exposure to cork infill and sands, while 79%, 71%, 68%, and 17% of MRSA remained viable after exposure to polymer-coated materials, crumb rubber, TPE rubber, and EPDM rubber, respectively. Our data also indicated that organic and sand infills exhibited minimal toxicity to MRSA, while high relative toxicity drives limited MRSA availability in EPDM rubber infill. MRSA recovery and viability varied among infill types but not among differing turf blades. The results of this study can inform appropriate athlete and field management practices.

Resurrection of antibiotics that methicillin-resistant Staphylococcus aureus resists by silver-doped bioactive glass-ceramic microparticles

This work describes a novel strategy to combat methicillin-resistant Staphylococcus aureus (MRSA) via the reactivation of inert antibiotics. This strategy exploits a multifunctional system consisting of bioactive glass–ceramic microparticles with antibacterial properties combined with various antibiotics to kill MRSA. Specifically, sol–gel derived silver-doped bioactive glass–ceramic microparticles (Ag-BG) combined with antibiotics that MRSA resists such as oxacillin or fosfomycin, significantly decreased the viability of MRSA. Ag-BG also potentiated the activity of vancomycin on static bacteria, which are typically resistant to this antibiotic. Notably, the synergistic activity is restricted to cell-envelope acting antibiotics as Ag-BG supplementation did not increase the efficacy of gentamicin. Bacteria viability assays and electron microscopy images demonstrate that Ag-BG synergizes to restore antibacterial activity to antibiotics that MRSA resists. The low cytotoxicity previously studied against oral bacteria, together with the known regenerative properties presented in previous studies, and the unique antibacterial properties observed in this work when they are combined with antibiotics, make this multifunctional system a promising approach for healing infected tissue. Statement of SignificanceThis study addresses a very significant issue in the field of antibiotic resistance presenting an innovative way to clear MRSA, by utilizing bioactive glass–ceramic microparticles in combination with antibiotics. Multifunctional glass–ceramic microparticles doped with silver ions (Ag-BG) have been previously observed to exhibit bioactive and antibacterial properties. In this study Ag-BG microparticles were observed to synergize with antibiotics restoring their sensitivity against MRSA. This research work presents a novel approach to resurrect ineffective antibiotics and render them effective against MRSA. Cytotoxicity to eukaryotic cells is not anticipated, as it has been previously observed that these microparticles can trigger hard and soft dental tissue regeneration, when they are utilized in certain concentrations. This study opens a new avenue in the treatment of multidrug resistance bacteria.

Targeted laser therapy synergistically enhances efficacy of antibiotics against multi-drug resistant Staphylococcus aureus and Pseudomonas aeruginosa biofilms

The growing prevalence of biofilm-associated multi-drug resistant (MDR) bacteria necessitates the innovation of non-traditional approaches to improve the effectiveness of mainstay antibiotics. Here, we evaluated the use of gold nanoparticle (GNP)-targeted pulsed laser therapy to enhance antibiotic efficacy against in vitro methicillin-resistant Staphylococcus aureus (MRSA) and MDR Pseudomonas aeruginosa biofilms. Treatment with antibody-conjugated GNPs followed by nanosecond-pulsed laser irradiation at 532 nm (~1.0 J/cm2) dispersed 96–99% of the biofilms relative to controls. GNP-targeted laser therapy combined with gentamicin or amikacin caused a synergistic 4- and 5-log reduction in the viability of MRSA and P. aeruginosa biofilms, respectively, whereas GNP-targeted laser therapy or antibiotics alone decreased biofilm viability by only ~1 log. Notably, GNP-targeted laser therapy was able to increase the antibiotic susceptibility of the biofilms to the level of drug sensitivity observed in planktonic MRSA and P. aeruginosa cultures, further indicating effective biofilm dispersal via this novel approach.

Peroxisome Proliferator-Activated Receptor γ Is Essential for the Resolution of Staphylococcus aureus Skin Infections

Skin/soft tissue infections (SSTIs) caused by methicillin-resistant Staphylococcus aureus (MRSA) represent serious healthcare burdens worldwide. The host initially controls these infections with a pro-inflammatory infiltrate. However, once established, MRSA viability remains constant. To clear established MRSA SSTIs, the host must transition into the post-inflammatory resolution phase marked by infiltration of alternatively activated macrophages. Here we show that the host nuclear receptor, peroxisome proliferation activator receptor γ (PPARγ), is essential for this transition and MRSA clearance. Chemical PPARγ inhibition or genetic ablation of PPARγ in myeloid cells results in an extended inflammatory phase and exacerbated MRSA SSTIs. Conversely, treating mice with PPARγ agonists hastens the onset of the resolution phase and improves MRSA clearance in a myeloid-dependent fashion. The resolving fibrotic abscess lacks abundant glucose and oxygen but is replete with antimicrobial peptides, which together contribute to MRSA clearance. Thus, PPARγ agonists may serve as viable treatment options for complicated MRSA SSTIs.

Cinnamaldehyde Inhibits MRSA Biofilm Formation and Reduces Cell Viability

The NIH identified that most microbial infections are biofilm-associated. Bacterial biofilm formation in human infection is of great concern to public health, as it has been associated with increased antimicrobial resistance, decreased effectiveness of host response, chronicity of infection, and medical device-associated disease. The pathogen, methicillin-resistant Staphylococcus aureus (MRSA), warrants special attention since it has been a frequent culprit in hospital- and community-acquired infections, is known to form biofilms in vivo, and is notoriously resistant to antimicrobics. This study sought to inhibit biofilm formation and/or reduce MRSA viability using the phytochemical cinnamaldehyde, which has been widely studied as an antimicrobial agent as well as a quorum sensing inhibitor. Clinical MRSA isolates from area hospital laboratories were assessed for cinnamaldehyde effect using a: (i) microplate assay for quantitative spectrophotometric evaluation of crystal violet-stained biofilm adherent to microwells; and (ii) viable bacterial count assay for colony forming unit (CFU/ml) enumeration. Results indicated that cinnamaldehyde inhibited MRSA biofilm formation in a concentration-dependent manner with significance (p ABBREVIATIONS:MRSA – methicillin-resistant S. aureus,MSSA – methicillin-susceptible S. aureus,TSB – tryptic soy broth

Inhibition of Neutrophil Adhesion and Antimicrobial Activity by Diluted Hydrosol Prepared from Rosa damascena.

Hydrosol prepared from the flowers of Rosa damascena (rose water) has been traditionally used for various health-related issues, including skin troubles such as erythema, itchiness, swelling. For the care of these skin troubles caused by microbial infection, both antimicrobial and antiinflammatory effects are required. Here, we investigated the effects of rose water on the growth of Candida albicans and methicillin-resistant Staphylococcus aureus (MRSA), which cause skin infections, and on the function of neutrophils, which play a major role in the regulation of inflammatory reactions. To assess its modulatory effects on neutrophils, the effects of rose water against neutrophil adhesion response were evaluated. Rose water inhibited mycelial growth of C. albicans at a concentration of ca. 2.2%, and reduced viability of MRSA within 1 h. Rose water suppressed neutrophil activation induced by lipopolysaccharide (LPS), tumor necrosis factor alpha (TNF-α), and N-formyl-Met-Leu-Phe (fMLP) at 5-15%. It also reduced the LPS- and TNF-α-induced cell surface expression of the adhesion-related molecule, cluster of differentiation (CD) 11b, but did not affect the migratory capacity of neutrophils with or without chemoattractant. These results suggest that rose water may reduce the pathogenicity of microbes, and attenuate neutrophil stimulation, which is involved in inflammatory responses. These findings suggest that rose water has a potential effect to inhibit skin inflammation caused by microbes.

The Mechanism Underlying the Antibacterial Activity of Shikonin against Methicillin-Resistant Staphylococcus aureus.

Shikonin (SKN), a highly liposoluble naphthoquinone pigment isolated from the roots of Lithospermum erythrorhizon, is known to exert antibacterial, wound-healing, anti-inflammatory, antithrombotic, and antitumor effects. The aim of this study was to examine SKN antibacterial activity against methicillin-resistant Staphylococcus aureus (MRSA). The SKN was analyzed in combination with membrane-permeabilizing agents Tris and Triton X-100, ATPase inhibitors sodium azide and N,N′-dicyclohexylcarbodiimide, and S. aureus-derived peptidoglycan; the effects on MRSA viability were evaluated by the broth microdilution method, time-kill test, and transmission electron microscopy. Addition of membrane-permeabilizing agents or ATPase inhibitors together with a low dose of SKN potentiated SKN anti-MRSA activity, as evidenced by the reduction of MRSA cell density by 75% compared to that observed when SKN was used alone; in contrast, addition of peptidoglycan blocked the antibacterial activity of SKN. The results indicate that the anti-MRSA effect of SKN is associated with its affinity to peptidoglycan, the permeability of the cytoplasmic membrane, and the activity of ATP-binding cassette (ABC) transporters. This study revealed the potential of SKN as an effective natural antibiotic and of its possible use to substantially reduce the use of existing antibiotic may also be important for understanding the mechanism underlying the antibacterial activity of natural compounds.

Antimicrobial effect of an ultrasonic levitation washer disinfector with silver electrolysis and ozone oxidation on methicillin‐resistant Staphylococcus aureus

Methicillin-resistant Staphylococcus aureus (MRSA) has rapidly emerged as a cause of severe and intractable skin infection. At present, there are no effective topical treatments, and infection or colonization by MRSA of the skin raises serious medical problems. We developed an ultrasonic levitation washer that generates silver ions (Ag(+)) and ozone (O3) to clean and sterilize medical devices. We report the effect of ultrasonic levitation (levitation) with Ag(+) and O3 on MRSA in vitro and in vivo. Antimicrobial effect against six MRSA strains of all agr types was examined under three in vitro conditions; cells floating in a water tank, cells infiltrating-, and cells forming a biofilm on an atelocollagen membrane. In the in vivo studies, we assayed the number of MRSA organisms that survived treatment on murine skin ulcers and evaluated the ulcer size. Levitation with Ag(+) dramatically decreased the survival of MRSA floating in a water tank. Levitation with Ag(+) and O3 significantly decreased the viability of MRSA that had infiltrated or formed a biofilm on atelocollagen membranes regardless of the level of biofilm production. In vivo studies showed that the number of MRSA on murine skin ulcers was significantly decreased when 15-min treatment was performed for 7 consecutive days and that the ulcer size was significantly decreased after the seventh treatment course. Levitation with Ag(+) and O3 may be a valuable tool for treating MRSA infestation of the skin and for accelerating wound healing.

Laser light combined with a photosensitizer may eliminate methicillin-resistant strains of Staphylococcus aureus

Methicillin-resistant Staphylococcus aureus (MRSA) is a major cause of hospital acquired infection throughout the world especially in wound and burn infections, pneumonia, septicaemia and endocarditis. We describe the effect of a HeNe laser in combination with a TBO dye on the viability of MRSA. A total of 34 isolates of S. aureus were obtained from 100 patients suffering from burns or wounds and from the nasal vestibulum of medical and nonmedical staff as carriers; eight isolates were methicillin-resistant. The isolates were exposed for 5, 10 and 15 min to a HeNe laser at a wavelength of 632.8 nm and 7.5 mW output power in the presence of 50 microg/ml toluidune blue O photosensitizer. The viable count was substantially decreased as determined by the plate count method for the three exposure times, with 100% killing with the 15-min exposure time. No significant effect was observed on MRSA isolates exposed to the laser alone. So MRSA was completely eradicated following 15 min exposure to a 632.8-nm HeNe laser in the presence of 50 microg/ml toluidune blue O photosensitizer under in vitro conditions.

Evaluation of the Copan ESwab transport system for the detection of methicillin-resistant Staphylococcus aureus: a laboratory and clinical study

Methicillin-resistant Staphylococcus aureus (MRSA) has emerged as an important pathogen. Successful detection starts with the selection of an appropriate specimen collection device. The Copan ESwab, a new swab, was compared with a commonly used Copan Venturi swab for the recovery of MRSA. In vitro assessment was performed according to the Clinical and Laboratory Standards Institute (CLSI) M40-A protocol. For the clinical evaluation, 24 patients with known MRSA carriage were included. The ESwab fulfilled the CLSI acceptance criteria for MRSA viability. In the clinical study, both swabs performed equally in qualitative terms of positive and negative. However, the in vitro and in vivo evaluation revealed at least 3.6-fold higher recovery of viable MRSA with the ESwab as compared with the Venturi swab. In conclusion, the ESwab may contribute to improve the quality of a MRSA screening protocol.

Garlic extract and two diallyl sulphides inhibit methicillin-resistant Staphylococcus aureus infection in BALB/cA mice.

The inhibitory effect of garlic extract, diallyl sulphide and diallyl disulphide against methicillin-resistant Staphylococcus aureus (MRSA) infection in BALB/cA mice was studied. The influence of these agents upon the levels of fibronectin, interleukin-6 and lipid oxidation in MRSA-infected mice was examined. Garlic extract at 100% and 50%; diallyl sulphide (DAS) at 10% and 5%; diallyl disulphide (DADS) at 1% and 0.5% were used in this study. Sixteen clinical MRSA isolates obtained from infected patients were used in this study (n=16). Mice were infected by injecting 200 microL MRSA-PBS solution, which contained 10(7) cfu, via the tail vein. At 16 h post-infection (p.i.), garlic extract, DAS or DADS at 200 microL was administrated orally. At 24 h p.i., mice were killed and blood, liver, kidney and spleen of each mouse were collected. Plasma and the filtrate from each organ and serial dilutions were used to determine colony count. Plasma fibronectin level was determined by rabbit anti-rat fibronectin antibody and quantified by ELISA. Interleukin-6 levels were determined by commercial kit. Lipid oxidation was determined by measuring malondialdehyde levels. The oral administration of these agents significantly decreased the viability of MRSA, in plasma, liver, kidney and spleen (P<0.05). MRSA infection significantly increased fibronectin and interleukin-6 levels in plasma of MRSA-infected mice (P<0.05); however, the oral administration of garlic extract and two diallyl sulphides significantly reduced both fibronectin and interleukin-6 levels (P<0.05). MRSA infection also significantly enhanced lipid oxidation in plasma and three organs (P<0.05). The treatments of garlic extract and two diallyl sulphides significantly decreased the malondialdehyde level and showed antioxidant protection (P<0.05). These data strongly supported the conclusion that garlic extract, diallyl sulphide and diallyl disulphide possessed multiple protective functions against MRSA infection, in which diallyl sulphide and diallyl disulphide could be considered as novel therapeutic agents for the treatment of MRSA infection.