Cystic and non-cystic fibrosis Pseudomonas aeruginosa isolates are not differentiated by the quorum-sensing signaling and biofilm production.

The capability for biofilm and quorum-sensing (QS) signaling production among Pseudomonas aeruginosa isolates were evaluated. A total of 231 isolates were recovered from sputa of cystic fibrosis (CF, n = 104) and non-CF (non-CF, n = 127) patients. One hundred ninety-seven (85.3%; 95% CI 80.1-89.3%) were biofilm producers and 157 (68%; 95% CI 61.7-73.6%) were weak QS-producing. No difference was observed between CF and non-CF isolates regarding the ability to produce biofilm and QS-signaling. Interestingly, the degree of QS production appears to be related to the degree of biofilm production. Thus, blocking QS pathways may be crucial in the prevention and treatment of biofilm-related infections.

Nutritional requirement among Pseudomonas aeruginosa isolates recovered from respiratory clinical specimens at a tertiary hospital from South of Brazil.

We screened 349 isolates of P. aeruginosa from cystic fibrosis (CF+) and non-cystic fibrosis (CF-) patients for the auxotrophy. Fourteen (4.0%) were auxotrophic and among them only one was recovered from CF-patient showing that this characteristic is strongly associated with cystic fibrosis. In total, a requirement for 5 different compounds (or combination) was verified and, of these, methionine was the most common single amino acid required. Only one auxotrophic isolate was no able to produce biofilm in vitro.

Identification, antimicrobial resistance and genotypic characterization of Enterococcus spp. isolated in Porto Alegre, Brazil.

In the past two decades the members of the genus Enterococcus have emerged as important nosocomial pathogens worldwide. In the present study, we evaluated the antimicrobial resistance and genotypic characteristics of 203 Enterococcus spp. recovered from different clinical sources from two hospitals in Porto Alegre, Rio Grande do Sul, Brazil. The species were identified by conventional biochemical tests and by an automated system. The genetic diversity of E. faecalis presenting high-level aminoglycoside resistance (HLAR) was assessed by pulsed-field gel electrophoresis of chromosomal DNA after SmaI digestion. The E. faecalis was the most frequent specie (93.6%), followed by E. faecium (4.4%). The antimicrobial resistance profile was: 2.5% to ampicillin, 0.5% to vancomycin, 0.5% teicoplanin, 33% to chloramphenicol, 2% to nitrofurantoin, 66.1% to erythromycin, 66.5% to tetracycline, 24.6% to rifampicin, 30% to ciprofloxacin and 87.2% to quinupristin-dalfopristin. A total of 10.3% of the isolates proved to be HLAR to both gentamicin and streptomycin (HLR-ST/GE), with 23.6% resistant only to gentamicin (HLR-GE) and 37.4% only to streptomycin (HLR-ST). One predominant clonal group was found among E. faecalis HLR-GE/ST. The prevalence of resistance among beta-lactam antibiotics and glycopeptides was very low. However, in this study there was an increased number of HLR Enterococcus which may be spreading intra and inter-hospital.

Feasible identification of Staphylococcus epidermidis using desferrioxamine and fosfomycin disks.

Coagulase-negative Staphylococcus spp. (CoNS) have emerged as predominant pathogens in hospital-acquired infections, as well as reservoirs of antimicrobial resistance, increasing the necessity of developing reliable methods for identification of the most frequent species. The aim of this study was to propose a simplified method for identification of Staphylococcus epidermidis. A total of 490 isolates of CoNS were identified by Bannerman's method. Taking into account distinct approaches for identification of S. epidermidis, among CoNS, we proposed the use of only two disks: desferrioxamine for the initial trial, and fosfomycin to match the final identification. Of the 320 isolates susceptible to desferrioxamine, Bannerman's method identified 238 S. epidermidis and 73 S. hominis, while we achieved identification of 239 S. epidermidis and 76 S. hominis. Compared to Bannerman's method, the method proposed here obtained a sensitivity of 99.5%, and had a positive predictor value of 99.2%. We also used a genotypic method for identification of S. epidermidis by polymerase chain reaction (PCR) targeting the tuf gene. In conclusion, the method proposed here has proved to be useful for the identification of S. epidermidis, the most frequent species of CoNS isolated from blood cultures in clinical microbiology laboratories.

Antibiotic resistance and molecular typing of Pseudomonas aeruginosa: focus on imipenem.

Susceptibility tests by disk diffusion and by E-test and molecular typing by macrorestriction analysis were performed to determine the relatedness of Pseudomonas aeruginosa isolates from three distinct hospitals. The resistance profile of 124 isolates to 8 antimicrobial agents was determined in three different hospitals, in Porto Alegre, Brazil. Frequencies of susceptibility ranged from 43.9% for carbenicillin to 87.7% for ceftazidime. Cross-resistance data of imipenem-resistant isolates indicated that most (70%) were also resistant to carbenicillin, although 30% remained susceptible to ceftazidime and cefepime. In general, susceptibility profiles were not able to determine relatedness among isolates of P. aeruginosa. On the other hand, molecular typing by macrorestriction analysis demonstrated high discriminatory power and identified 66 strains among 72 isolates of P. aeruginosa. Imipenem-susceptible isolates were all different. However, identical clones of imipenem-resistant isolates were found in two of the hospitals, despite variable response to other antibiotics. No clustering of infection among the different medical centers was observed. In conclusion, clones of P. aeruginosa did not spread among the different hospitals in our city even though related isolates of imipenem-resistant P. aeruginosa were found.