The present study introduced T. daenensis and S. hortensis EOs as new antibiofilm, and S. hortensis EO as anti-QS herbal agents with natural origin against Staph. aureus.

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Cuminum cyminum L. (cumin) is valued for its aromatic and medicinal properties. There are several reports of antibacterial activity of C. cyminum essential oil (CcEO). Accordingly, the present study was conducted to investigate the mechanism(s) of action of the CcEO against multidrug-resistant (MDR) Staphylococcus aureus. Therefore, 10 S. aureus MDR isolates, obtained from different sources, were selected based on the antibiotic susceptibility patterns and the Clinical and Laboratory Standards Institute definition and subjected to the examinations. Our results exhibited promising bacteriostatic and bactericidal properties of the CcEO. The minimum inhibitory concentration (MIC) and the minimum bactericidal concentration values ranged from 5 to 10 and 10 to 20 μL ⋅ mL–1, respectively. Scanning electron microscope was used to assess the bacterial cell structure and morphology after the induction with 1/2 MIC concentration of the CcEO. The observed morphological changes appeared to be deformation of the cell membrane and destruction of the cells. In the case of quorum sensing inhibitory potential, treatment of S. aureus isolates with the sub-MIC concentrations (1/2 MIC) of the CcEO significantly reduced the hld expression (3.13-fold downregulation), which considerably controls S. aureus quorum-sensing accessory regulator system. Another virulence factor influenced by the CcEO was the polysaccharide intercellular adhesion production system, as an important component of cell–cell adhesion and biofilm formation. Consequently, the expression level of the intercellular adhesion (ica) locus in the S. aureus cells was examined following treatment with CcEO. The results showed significant decrease (−3.3-fold) in ica expression, indicating that the CcEO could potentially interfere with the process of biofilm formation. Using the ethidium bromide efflux inhibition assay, the S. aureus NorA efflux pump was phenotypically but not genotypically (in quantitative polymerase chain reaction assay) affected by the CcEO treatment. Using gas chromatography–mass spectrometry analysis, cuminic aldehyde (38.26%), α,β-dihydroxyethylbenzene (29.16%), 2-caren-10-al (11.20%), and γ-terpinene (6.49%) were the most detected compounds. The antibacterial and antivirulence action of the CcEO at sub-MIC concentrations means that no microbial resistance will be promoted and developed after the treatment with this agent. These findings revealed that the CcEO is a promising antibacterial agent to control infections caused by the MDR S. aureus strains.

Significance and Impact of the Study: We investigated the antibacterial, anti-biofilm and quorum sensing inhibitory properties of Thymus daenensis and Satureja hortensis essential oils against food-borne pathogen Escherichia coli O157:H7 using phenotypic and genotypic methods. Accordingly, the present study introduces T. daenensis and S. hortensis essential oils as new antimicrobial agents against E. coli O157:H7.

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Background: Despite the clinical importance of ventilator-associated pneumonia (VAP) as the most common nosocomial infection in ICU, there are few studies in Iran evaluating the bacterial causative agents involving VAP. Objectives:The aim of the present study was to determine the prevalence of bacterial agents of VAP, and to evaluate the presence of S. pneumoniae in VAP-confirmed ICU patients by real-time PCR. Methods: In this cross-sectional study, during March 2016 to March 2017, 50 tracheal aspirates were collected from VAP-confirmed ICU admitted patients in Tehran. The number of epithelial cells and white blood cells (WBC) were determined by direct microscopy. Bacterial identification from VAP samples was done by routine biochemical tests and culturing on differential media. DNA was extracted from samples, and based on lytA gene amplification, a quantitative real-time PCR was performed for S. pneumoniae detection and quantification. Results: In culture, a pure bacterium was isolated from 40 out of 50 samples (80%), with Klebsiella pneumoniae (26%) and Acinetobacter ssp. (18%) being the most common isolates, respectively; however, all cultures were negative for S. pneumoniae. By real-time PCR, two samples (4%) were positive for S. pneumoniae with 4×10 4 and 1.6×10 5 CFU/mL bacterial load. These two samples contained 10 and 12 WBC/lpf, respectively. Conclusions: Although with only a with few clinical samples, this is the first study reporting pneumococcal VAP in Iran. Furthermore, in regards to the importance of VAP in ICU patients, more studies to optimize cultural method and evaluate applicable diagnostic molecular methods could be appreciated.

Streptococcus pneumoniae (pneumococcus) is recognized as one of the major cause of infections in communities and hospitals. In this study, anti-pneumococcal and anti-efflux pump activity of two medicinal plants (Thymus daenensis and Origanum vulgare) essential oils were evaluated. Checkerboard assay test was performed for investigation of the effects of selected EOs on ciprofloxacin (CIP) and ethidium bromide (EtBr) uptake in pmrA-overexpressed fluoroquinolone-resistant pneumococcus. Using quantitative real-time RT-PCR the PmrA efflux pump gene (pmrA) expression was evaluated following treatment with selected EOs. Gas chromatography-mass spectrometry analysis was performed for identifying the major components of the tested EOs. The minimum inhibitory concentration (MIC) values for pneumococcus isolates were 0·625-2·5 μl ml for T. daenensis and 1·25-5 μl ml for O. vulgare EOs. We confirmed that in all strains T. daenensis and O. vulgare have a total or partial synergistic effects with CIP and EtBr (FICI from 0·14 to 0·75). In other hand MIC/2 concentration of T. daenensis and O. vulgare EOs caused a significant downregulation of pmrA gene (P < 0·05) in seven of eight strains. This study showed that T. daenensis and O. vulgare EOs have strong antimicrobial and anti-efflux pump activity against clinical isolates of S. pneumoniae and might be useful in controlling pneumococcal infections. SIGNIFICANCE AND IMPACT OF THE STUDY: This study introduced Thymus daenensis and Origanum vulgare essential oil as new antibacterial and anti-efflux pump agents against fluoroquinolone-resistant Streptococcus pneumoniae clinical isolates. These findings indicate that combination of these two essential oils with fluoroquinolone antibiotics may provide alternative methods to overcome the fluoroquinolone-resistant S. pneumoniae.

The aim of this study was to investigate the effect of Thymus daenensis L., Satureja hortensis L., and Origanum vulgare L. essential oils (EOs) on the planktonic growth, biofilm formation, quorum sensing (QS), and competence system (CS) of Streptococcus pneumoniae. The anti-biofilm activity of EOs was determined by Microtiter-Plate Test (MtP) and scanning electron microscope (SEM). The QS and CS inhibitory activities were determined on the pre-grown biofilm by gene expression analysis using quantitative real-time RT-PCR. Using gas chromatography-mass spectrometry analysis, the major components of the tested EOs were detected. The MtP and SEM detected a significant inhibitory effect of the three EOs on biofilm formation at sub-minimum inhibitory concentrations (MICs). The most anti-biofilm activity was seen for T. daenensis. LuxS and pfs genes (genes involved in QS) downregulated the following treatment with MIC/2 of Thymus and Satureja EOs. Thymol, carvacrol, p-cymene, pulegone, and 1,8-cineole were the major components of the tested EOs. The used EOs seem to be good candidates for preventing biofilm formation and subsequent colonization of S. pneumoniae. This study introduced T. daenensis and S. hortensis as new anti-biofilm and QS inhibitor agents with a natural origin.