Polyclonal outbreak of bacteremia caused by Burkholderia cepacia complex and the presumptive role of ultrasound gel.

A nosocomial polyclonal outbreak associated to bacteremia caused by different Burkholderia cepacia complex (BCC) species and clones is reported. Molecular characterization identified Burkholderia stabilis, Burkholderia contaminans, and Burkholderia ambifaria among BCC isolates obtained from patients in neonatal and adult intensive care units. BCC was also isolated from an intrinsically contaminated ultrasound gel, which constituted the presumptive BCC source. Prior BCC outbreak related to contaminated ultrasound gels have been described in the setting of transrectal prostate biopsy. Outbreak caused strains and/or clones of BCC have been reported, probably because BCC are commonly found in the natural environment; most BCC species are biofilm producers, and different species may contaminate an environmental source. The finding of multiple species or clones during the analysis of nosocomial BCC cases might not be enough to reject an outbreak from a common source.

Comparative time-kill study of doxycycline, tigecycline, cefazolin and vancomycin against several clones of Staphylococcus aureus.

BACKGROUND: We present herein, a comparative study assessing the bactericidal kinetics of tigecycline, doxycycline, cefazolin and vancomycin against several methicllin-susceptible (MSSA) and -resistant (MRSA) Staphylococcus aureus isolates recovered from patients of 24 different cities in Argentina. METHODS: After genotypic characterization, 20 strains (10 MRSA and 10 MSSA) were selected for time-kill studies. RESULTS: Vancomycin showed bactericidal effect (i.e. ≥3-log(10) CFU/mL decrease) against 50% and 10% of the MRSA strains at 4 x Minimal Inhibitory Concentration (MIC) and 2xMIC, respectively, after 24 h of incubation and displayed bactericidal activity against all MSSA isolates at 4xMIC. Cefazolin was bactericidal against 30% of MSSA strains at the higher concentration (4xMIC) and against 10% at 2 x MIC and MIC dose concentrations. The bactericidal magnitude of cefazolin observed after 24 h of incubation was lower than the vancomycin one. Albeit bacteriostactic, tigecycline at 2xMIC exerted a -1 to2-log decrease in the viable cell counts after 24-h incubation against 19 of the 20 S. aureus strains. Doxycycline was the least inhibitory of the antibiotics tested against both MRSA and MSSA, displaying no bactericidal activity in any of the cases and showing regrowth after 24 h of incubation at MIC level. CONCLUSION: Vancomycin at high concentrations showed the best activity. Cefazolin did not show the activity expected for a beta-lactam antibiotic against MSSA. Tigecycline may be a useful option in infections caused by MRSA, where bactericidal activity is not an exclusive requirement and doxycycline does not seem an attractive alternative in serious infections.

In vitro susceptibility of Achromobacter spp. isolates: comparison of disk diffusion, Etest and agar dilution methods.

In this study, we analysed the antimicrobial susceptibility of 92 strains of Achromobacter spp. isolated from clinical samples to 18 antimicrobial agents. The disk diffusion method and Etest were compared with the agar dilution method, and the breakpoints of susceptibility and resistance for the disk diffusion method for the antimicrobials tested were determined. The most active antibiotics were piperacillin, piperacillin/tazobactam and the carbapenems. By applying the linear least-squares regression method, breakpoints could be established for antibiotics active against this genus such as imipenem, meropenem, ertapenem and trimethoprim/sulfamethoxazole (SXT). Other active antibiotics, such as piperacillin and minocycline, could be tested by the Etest method. The less active antibiotics such as gentamicin, doxycycline and tetracycline could be tested by the disk diffusion method. For the rest of the antimicrobial agents tested, breakpoints could not be established owing to the high percentage of errors and/or the poor linear regression coefficient obtained. Therefore, these antimicrobial agents should be tested by minimal inhibitory concentration determination. In summary, we recommend the following zone diameter breakpoints for resistant and susceptible, respectively: < or = 11 mm and > or = 22 mm for imipenem; < or = 13 mm and > or = 24 mm for meropenem; < or = 17 mm and > or = 24 mm for ertapenem; < or = 15 mm and > or = 21 mm for gentamicin; < or = 27 mm and > or = 28 mm for SXT; < or = 20 mm and > or = 29 mm for tetracycline; and < or = 20 mm and > or = 24 mm for doxycycline.

A convenient microbiological assay employing cell-free extracts for the rapid characterization of Gram-negative carbapenemase producers.

OBJECTIVES: The dissemination of metallo and serine carbapenem-hydrolysing beta-lactamases among Gram-negative nosocomial bacteria represents an acute problem worldwide. Here, we present a rapid and sensitive assay for the characterization of carbapenemase producers to aid in infection control and prevention. METHODS: The assay involves a rapid disruption of bacterial isolates with silicon dioxide microbeads, followed by the testing in cell-free extracts of hydrolytic activity towards various beta-lactams including two carbapenems (imipenem and meropenem) and a cephalosporin (ceftazidime). A parallel testing of the effects of selective beta-lactamase inhibitors such as EDTA and clavulanic acid allows differentiation of metallo carbapenemases from serine carbapenemases, and also clavulanic-acid-sensitive from -resistant enzymes among the latter. RESULTS: The efficiency of bacterial disruption using silicon dioxide microbeads was identical to that of ultrasonic treatment. The subsequent microbiological assay aimed to evaluate both substrate specificity and inhibitor profile of carbapenem-hydrolysing enzymes present in the extracts and allowed an accurate differentiation of A, B and D types, as judged by the analysis of 24 well-characterized clinical strains that included metallo-beta-lactamase producers (i.e. VIM-, IMP- and SPM-type Pseudomonas producers; an L1 Stenotrophomonas maltophilia producer; and a GOB-18 Elizabethkingia meningoseptica producer) as well as serine carbapenemase producers (i.e. an SME-type Serratia marcescens producer, a GES-2 Pseudomonas aeruginosa producer, Klebsiella pneumoniae and Citrobacter freundii KPC-2 producers and OXA-type Acinetobacter baumannii producers). CONCLUSIONS: We have developed a convenient microbiological assay aimed to more accurately and in a short time characterize carbapenem-hydrolysing enzymes produced by Gram-negative bacteria. The assay possesses broad applicability in the clinical setting.

Comparative time-kill study of doxycycline, tigecycline, sulbactam, and imipenem against several clones of Acinetobacter baumannii.

To assess potential alternative options for the treatment of infections caused by Acinetobacter baumannii, we performed time-kill studies of doxycycline and tigecycline using several isolates recovered from patients residing in 10 different cities in Argentina. Imipenem and sulbactam were also included for comparison purposes. Eleven isolates representing 5 distinctive clones, or isolates with different susceptibility patterns within the same clone, were selected. Tubes containing cation-supplemented Mueller-Hinton broth with and without antibiotics were seeded with a log-phase inoculum of roughly 5 x 10(5) CFU/mL. By using the viable counts determined at 2-, 4-, 6-, 8-, and 24-h intervals after inoculation, a 24-h time-kill curve was constructed for each isolate. No bactericidal activity (defined as a >or=3-log(10) CFU/mL decrease in the viable cell counts with respect to the original inoculum) was observed at any time with sulbactam (4 microg/mL) or tigecycline (1 microg/mL), whereas low bactericidal rate (18% of the isolates) was shown for doxycycline (1 microg/mL) and sulbactam (16 microg/mL) after 24 h of incubation. Doxycycline (4 microg/mL) and tigecycline (8 microg/mL) displayed bactericidal activity at 24 h of incubation against 36% and 54% of the isolates, respectively, including the carbapenem-resistant isolate. Corresponding values for imipenem (1 and 4 microg/mL) against the 10 carbapenem-susceptible isolates were 60% and 90%, respectively. The present study confirms the in vitro efficacy of imipenem against A. baumannii, suggests that doxycycline could be a suitable, cost-effective, alternative option in some instances, and sheds light on the potential role of tigecycline in the treatment of infections with this organism.

Sensitive EDTA-based microbiological assays for detection of metallo-{beta}-lactamases in nonfermentative gram-negative bacteria.

The worldwide spread of metallo-beta-lactamase (MBL)-producing gram-negative bacilli represents a great concern nowadays. Sensitive assays for their specific detection are increasingly demanded to aid infection control and to prevent their dissemination. We have developed a novel microbiological assay employing crude bacterial extracts, designated EDTA-imipenem microbiological assay (EIM), to identify MBLs in nonfermentative gram-negative clinical strains. We also evaluated the ability of EIM to detect MBLs in comparison to those of other currently employed screening methods, such as the EDTA disk synergy test (EDS) with imipenem as a substrate and the Etest method. The sensitivities of EIM and Etest were similar (1 versus 0.92, respectively) and much higher than that of EDS (0.67). Moreover, both EIM and Etest displayed the maximum specificity. Modifications were introduced to EDS, including the simultaneous testing of three different beta-lactams (imipenem, meropenem, and ceftazidime) and two different EDTA concentrations. This resulted in a sensitivity improvement (0.92), albeit at a cost to its specificity. A simple strategy to accurately detect MBL producers is proposed; this strategy combines (i) an initial screening of the isolates by the extended EDS assay to select the potential candidates and (ii) confirmation of the true presence of MBL activity by EIM.