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Enterococcus faecalis and Enterococcus faecium are among the main agents associated with nosocomial infections with high mortality in immunocompromised patients. Antibiotic resistance, especially against gentamicin and vancomycin among Enterococci, is a risk factor that could increase the morbidity and mortality rate. 179 Enterococci isolates from burn patients were included in this study. Antibiotic susceptibility testing was done using the disk diffusion test and minimum inhibitory concentration (MIC) was evaluated by agar microdilution. Vancomycin and gentamicin resistance associated genes including vanA, vanB, vanC, aac (6’)-Ie aph(2’’), aph(3’)-IIIa and ant(4’)-Ia were detected by PCR and their statistical relation with antibiotic resistance was evaluated. E. faecalis was the more prevalent strain among our local isolates and showed a higher antibiotic resistance in comparison to E. faecium. Vancomycin had a good antibacterial effect on the Enterococcus spp. isolates; however, resistance to this antibiotic and a high-level gentamicin resistance (HLGR) phenotype were observed. Among van operon genes, vanA was the most prevalent gene and among the gentamicin resistance genes, aph (3’)-IIIa was more frequent. The HLGR Enterococci are a real challenge in nosocomial infections. Vancomycin is a key antibiotic to treat such infections but emergence of VRE in our region could be a real concern and, therefore, phenotypic and molecular surveillance must be considered.
Pheromone-responsive plasmids constitute a unique group of ~ 20 plasmids identified, as yet, only among enterococcal species. Several of their representatives, e.g. pAD1, pCF10, pPD1 and pAM373 have been extensively studied. These plasmids posses a sophisticated conjugation mechanism based on response to sex pheromones - small peptides produced by plasmid-free recipient cells. Detailed analysis of regulation and function of the pheromone response process revealed its great complexity and dual role - in plasmid conjugation and modulation of enterococcal virulence. Among other functional modules identified in pheromone plasmids, the stabilization/partition systems play a crucial role in stable maintenance of the plasmid molecule in host bacteria. Among them, the par locus of pAD1 is one of the exceptional RNA addiction systems. Pheromone-responsive plasmids contribute also to enterococcal phenotype being an important vehicle of antibiotic resistance in this genus. Both types of acquired vancomycin resistance determinants, vanA and vanB, as well many other resistant phenotypes, were found to be located on these plasmids. They also encode two basic agents of enterococcal virulence, i.e. aggregation substance (AS) and cytolysin. AS participates in mating-pair formation during conjugation but can also facilitate the adherence of enterococci to human tissues during infection. The second protein, cytolysin, displays hemolytic activity and helps to invade eukaryotic cells. There are still many aspects of the nature of pheromone plasmids that remain unclear and more detailed studies are needed to understand their uniqueness and complexity.
The activity of ciprofloxacin, sparfloxacin and moxifloxacin was determined for 205 Enterococcus faecalis isolates from patients of five hospitals (Warsaw, Poland; collected from 2000 to 2002). Ciprofloxacin resistant and intermediate isolates were numerous (53.7%). Among them, highly resistant (MIC≥16 mg/l) isolates predominated (98%). Isolates resistant to ciprofloxacin were also resistant to sparfloxacin and moxifloxacin. The parC and gyrA QRDRs (quinolone-resistance-determining region) of 11 isolates with ciprofloxacin MICs from 1 to 256 mg/l were analysed by DNA sequencing. In ParC one kind of amino acid substitution (of Ser-85 to Ile) in 9 E. faecalis strains with MICs from 16 to 256 mg/l was observed. In GyrA Ser-84 was changed to one of four different amino acids: Arg, He, Cys or Tyr, however no association between the amino acid type and MIC value was found. The last two substitutions have not been reported to date for E. faecalis. Moreover, our results may suggest that mutations within parC and gyrA are associated with development of a high-level of ciprofloxacin resistance.
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