Lenka Černohorská

Antibiotic synergy against biofilm-forming Pseudomonas aeruginosa

Eight antibiotics (aztreonam, ceftazidim, cefoperazon, cefepim, netilmicin, amikacin, ofloxacin and ciprofloxacin) exhibited antimicrobial activity individually and/or in combinations against 20 wild-type biofilm-forming strains of Pseudomonas aeruginosa. The strains were less susceptible in biofilm; in 10 strains antibiotic synergy was observed for the combination of aztreonam and ciprofloxacin. Synergy was also demonstrated in the case of β-lactams and aminoglycosides, β-lactams and fluoroquinolones, aminoglycosides and fluoroquinolones, and for monobactams and β-lactams although the strains were resistant to the individual antibiotics. Synergism or partial synergism was found with one or more antibiotic combinations against 32.4% of isolates.

Determination of minimal regrowth concentration (MRC) in clinical isolates of various biofilm-forming bacteria

Based on the ability to attach to polymeric surfaces, the formation of biofilms was determined in 5 wild-type strains (Pseudomonas aeruginosa, Klebsiella pneumoniae, Acinetobacter baumanii, Escherichia coli, Staphylococcus warneri). Using modified Christensen method, minimum regrowth concentration (MRC) of piperacillin, piperacillin-tazobactam, cefoperazon, ceftazidim, cefepim, meronem, ciprofloxacin, netilmicin and amikacin for Gram-negative and of ampicillin-sulbactam, chloramphenicol, tetracycline, clindamycin, vancomycin and teicoplanin for Gram-positive bacteria was estimated in trypticase-soy broth medium after a 1-d growth on polystyrene microtiter plates. Adherent bacterial populations exhibited reduced antimicrobial susceptibility, which was not shown in submerged cultures. Our results indicate that MRC can predict therapeutic outcome of antibiotic treatment better than the minimum inhibitory concentration tests commonly used.

Characterization of Human Uropathogenic ESBL-Producing Escherichia coli in the Czech Republic: Spread of CTX-M-27-Producing Strains in a University Hospital

AIMS The purpose of this study was to characterize the extended-spectrum β-lactamase (ESBL)-producing uropathogenic Escherichia coli (UPEC) strains isolated in the South Moravia region of the Czech Republic. RESULTS Out of 109 ESBL-producing UPEC isolates, the CTX-M-15-producing E. coli O25b-ST131 was detected in 55 (50.5%) and the CTX-M-27-producing E. coli O25b-ST131 in 40 isolates (36.7%). Most isolates were distributed among three pulsed-field gel electrophoresis clusters and were characterized by low variability relative to antibiotic resistance patterns, in E. coli phylogroups and by the prevalence of virulence and bacteriocin determinants. Despite this, 14 groups of identical isolates (comprising a total of 41 isolates) were identified when all tested parameters of E. coli were combined. CONCLUSIONS Since the occurrence of E. coli B2-O25b-ST131 CTX-M-27 was only recently described in Asia, the frequent isolation of this lineage among patients in South Moravia suggests an efficient transfer of this clone from Asian countries. The limited variability of detected parameters of ESBL-producing UPEC strains is consistent with a common origin of the analyzed isolates, in which there is an ongoing process of genetic diversification.

Phenotypes of Escherichia coli isolated from urine: Differences between extended-spectrum β-lactamase producers and sensitive strains

BACKGROUND Escherichia coli is a frequent causative agent of urinary tract infections, and increasing resistance of E. coli to antimicrobials presents a growing challenge. METHODS Here we compare phenotypes of extended-spectrum β-lactamase (ESBL) producers (n = 220) with a control group of sensitive strains (non-ESBL producers; n = 150). For each strain, we assessed the presence of O25 antigen, hemolysis, biofilm production, sensitivity to antibiotics, and biochemical profile. RESULTS Compared to the control group, ESBL producers were more frequently O25 positive (6.0% vs. 42.3%) and less frequently hemolytic (34.7% vs. 6.4%). Comparison of biofilm production in brain-heart infusion (BHI) and in BHI with 4% glucose supplementation showed that ESBL-positive strains produced biofilm in BHI with glucose less intensely than the control group (p < 0.05). Most ESBL producers were ciprofloxacin-resistant (91.8%). Biochemical analyses revealed that ESBL producers more frequently utilized inositol, ornithine, sorbitol, melibiose, and saccharose, whereas the control group more frequently used esculin, lysine, arginine, and dulcitol. The control group strains with O25 antigen were more commonly resistant to ciprofloxacin (p < 0.05). Pulsed-field gel electrophoresis results showed higher variability among the control group of sensitive strains. CONCLUSION These findings suggest a potential to detect ESBL strains based on virulence factors and biochemical properties, which could be useful in shaping proper empiric antimicrobial therapy, and for initiating such therapy as soon as possible.