Satureja montana L. essential oil and its antimicrobial activity alone or in combination with gentamicin.

Many essential oils (EOs) are screened as potential sources of antimicrobial compounds. EOs from the genus Satureja have recognized biological properties, including analgesic, anti-inflammatory, immunomodulatory, anticancer, and antimicrobial activity. This study aimed to obtain a metabolite profile of commercial essential oil of S. montana L. (SEO) and to evaluate its antimicrobial properties, both alone and combined with gentamicin towards Gram-negative and Gram-positive bacterial strains. Untargeted analyses based on direct infusion Fourier-transform ion cyclotron resonance mass spectrometry (FT-ICR MS) and on GC-MS have provided a high metabolome coverage, allowing to identify carvacrol, cymene and thymol as the major components of commercial SEO. SEO exerted an antimicrobial activity and induced a synergistic interaction with gentamicin against both reference and clinical bacterial strains. A significant reduction of Escherichia coli, Staphylococcus aureus and Listeria monocytogenes biofilm formation was induced by SEO. As a result of SEO treatment, clear morphological bacterial alterations were visualized by scanning electron microscopy: L. monocytogenes and S. aureus showed malformed cell surface or broken cells with pores formation, whereas E. coli displayed collapsed cell surface. These results encourage further studies about bactericidal and antibiotic synergistic effect of SEO for combined therapy in clinical setting as well as in agricultural systems.

Rapid Detection of Antibiotic Resistance in Gram-Negative Bacteria Through Assessment of Changes in Cellular Morphology.

Rapid antimicrobial susceptibility testing has the potential to improve patient outcomes and reduce healthcare-associated costs. In this study, a novel assay based on bacterial cell elongation after exposure to an antibiotic (ceftazidime) was evaluated for its ability to rapidly detect resistance in Gram-negative bacteria. The assay was used to detect resistance in a large collection of strains containing 320 clinical isolates of Acinetobacter baumannii, 171 clinical isolates of Klebsiella pneumoniae, and 212 clinical isolates of Pseudomonas aeruginosa, and the results were compared to those obtained using standard antimicrobial susceptibility testing methods. The assay identified ceftazidime-resistant strains with 100% sensitivity and 100% specificity for A. baumannii, 100% sensitivity and 97.2% specificity for K. pneumoniae, and with 82.3% sensitivity and 100% specificity for P. aeruginosa. Importantly, results were obtained in 1 hour 15 minutes from exponentially growing cultures. This study demonstrates that changes in cell length are highly correlated with phenotypic antibiotic susceptibility determined using standard susceptibility testing methods. This study therefore provides proof-of-concept that changes in cell morphology can be used as the basis for rapid detection of antibiotic resistance and provides the basis for the development of novel rapid diagnostics for the detection of antibiotic resistance.

The efflux inhibitor phenylalanine-arginine beta-naphthylamide (PAßN) permeabilizes the outer membrane of gram-negative bacteria.

Active efflux of antimicrobial agents is a primary mechanism by which bacterial pathogens can become multidrug resistant. The combined use of efflux pump inhibitors (EPIs) with pump substrates is under exploration to overcome efflux-mediated multidrug resistance. Phenylalanine-arginine ß-naphthylamide (PAßN) is a well-studied EPI that is routinely combined with fluoroquinolone antibiotics, but few studies have assessed its utility in combination with ß-lactam antibiotics. The initial goal of this study was to assess the efficacy of ß-lactams in combination with PAßN against the opportunistic pathogen, Pseudomonas aeruginosa. PAßN reduced the minimal inhibitory concentrations (MICs) of several ß-lactam antibiotics against P. aeruginosa; however, the susceptibility changes were not due entirely to efflux inhibition. Upon PAßN treatment, intracellular levels of the chromosomally-encoded AmpC ß-lactamase that inactivates ß-lactam antibiotics were significantly reduced and AmpC levels in supernatants correspondingly increased, potentially due to permeabilization of the outer membrane. PAßN treatment caused a significant increase in uptake of 8-anilino-1-naphthylenesulfonic acid, a fluorescent hydrophobic probe, and sensitized P. aeruginosa to bulky antibiotics (e.g. vancomycin) that are normally incapable of crossing the outer membrane, as well as to detergent-like bile salts. Supplementation of growth media with magnesium to stabilize the outer membrane increased MICs in the presence of PAßN and restored resistance to vancomycin. Thus, PAßN permeabilizes bacterial membranes in a concentration-dependent manner at levels below those typically used in combination studies, and this additional mode of action should be considered when using PAßN as a control for efflux studies.

Studies on the biocidal and cell membrane disruption potentials of stem bark extracts of Afzelia africana (Smith).

We had recently reported antibacterial activity in the crude extract of the stem bark of Afzelia africana (Akinpelu et al., 2008). In this study, we assessed the biocidal and cell membrane disruption potentials of fractions obtained from the crude extract of the plant. The aqueous (AQ) and butanol (BL) fractions exhibited appreciable antibacterial activities against the test bacteria. The minimum inhibitory concentrations of the AQ and BL fractions ranged between 0.313 and 2.5 mg/ml, while their minimum bactericidal concentrations varied between 0.625 and 5.0 mg/ml. Also, the AQ fraction killed about 95.8% of E. coli cells within 105 min at a concentration of 5 mg/ml, while about 99.1% of Bacillus pumilus cells were killed by this fraction at the same concentration and exposure time. A similar trend was observed for the BL fraction. At a concentration of 5 mg/ml, the butanol fraction leaked 9.8 microg/ml of proteins from E. coli cells within 3 h, while the aqueous fraction leaked 6.5 microg/ml of proteins from the same organisms at the same concentration and exposure time. We propose that the stem bark of Afzelia africana is a potential source of bioactive compounds of importance to the pharmaceutical industry.

Studies on the biocidal and cell membrane disruption potentials of stem bark extracts of Afzelia africana (Smith)

We had recently reported antibacterial activity in the crude extract of the stem bark of Afzelia africana (Akinpelu et al., 2008). In this study, we assessed the biocidal and cell membrane disruption potentials of fractions obtained from the crude extract of the plant. The aqueous (AQ) and butanol (BL) fractions exhibited appreciable antibacterial activities against the test bacteria. The minimum inhibitory concentrations of the AQ and BL fractions ranged between 0.313 and 2.5 mg/ml, while their minimum bactericidal concentrations varied between 0.625 and 5.0 mg/ml. Also, the AQ fraction killed about 95.8 percent of E. coli cells within 105 min at a concentration of 5 mg/ml, while about 99.1 percent of Bacillus pumilus cells were killed by this fraction at the same concentration and exposure time. A similar trend was observed for the BL fraction. At a concentration of 5 mg/ml, the butanol fraction leaked 9.8 μg/ml of proteins from E. coli cells within 3 h, while the aqueous fraction leaked 6.5 μg/ml of proteins from the same organisms at the same concentration and exposure time. We propose that the stem bark of Afzelia africana is a potential source of bioactive compounds of importance to the pharmaceutical industry.

[Effect of water pollution by oil and oil products on barrier functions of bacterial cell cytoplasmic membranes]. / Vliianie zagriazneniia vodnoi sredy neft'iu i nefteproduktami na bar'ernye svoistva tsitoplazmaticheskikh membran bakterial'nykh kletok.

The effects of oil, diesel fuel, and kerosene on the electroorientational spectra and osmo-optical characteristics of bacterial cells were studied. Electroorientational spectra were found to be affected over the entire frequency range studied; changes in low-frequency (< 100 kHz) electroorientation were related to alterations in the cell surface, and those in high-frequency electroorientation, to the impairment of the barrier function of the plasma membrane. The membranotropic activity of petroleum products was also demonstrated by the osmo-optical method. Of nine bacterial species studied, Pseudomonas fluorescens VKM B-894, P. oleovorans VKM B-1522, and P. stutzeri VKM B-903 were most susceptible to the membranotropic action of kerosene; P. putida VKM B-1292 was the most resistant. Other bacterial strains studied were moderately sensitive.

[Spheroplast formation in clinical isolates of gram-negative bacteria by beta-lactam antibiotic in the presence of Ca2+ or Mg2+].

Induction to cell wall-deficient bacteria has been suggested to be a cause of intractable and opportunistic infection after chemotherapy. Spheroplast formation by beta-lactam antibiotic in not so high osmotic pressure was investigated in six species of gram-negative bacteria. Some species of gram-negative bacteria were induced to form spheroplast at a high rate by 1:10 concentration of ceftizoxime in the presence of Ca2+ or Mg2+. Especially in 38% of Proteus mirabilis and P. vulgaris, more than 40% of the original cells were induced to form spheroplast by ceftizoxime in a medium supplemented with 40 mM Ca2+. The same formation rate was also found in 22% of Serratia marcescens. Formation rates in the drug sensitive strains of S. marcescens were enhanced as the drug concentration increased. Ca2+ was more effective in spheroplast formation than Mg2+.