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We evaluated several EDTA-based combined-disk tests to detect 25 mcr producers among 48 Enterobacteriaceae. Colistin disks plus EDTA (292/584 μg) on MH and CAMH agar were used. Results were positive if with chelator there was an inhibition zone increase ≥3 mm compared to colistin alone. All tests resulted unreliable (sensitivity ≤68%)., (Copyright © 2018 Elsevier B.V. All rights reserved.)

The in-house Carba-NP and Blue-Carba tests were compared using 30 carbapenemase- and 33 non-producing Enterobacteriaceae. Tests were read by three operators. 100% sensitivity was reported for both tests, but Carba-NP was slightly more specific than Blue-Carba (98.9% vs. 91.7%). We describe potential sources of error during tests' preparation and reading., (Copyright © 2016 Elsevier B.V. All rights reserved.)

The aim of this study was to characterize the resistance mechanisms and genetic relatedness of 21 carbapenem-resistant Acinetobacter baumannii blood isolates collected in Italy during a 1-year multicenter prospective surveillance study. Genes coding for carbapenemase production were identified by polymerase chain reaction (PCR) and sequencing. Pulsed-field gel electrophoresis (PFGE), multiplex PCRs for group identification, and multilocus sequence typing (MLST) were used to determine genetic relationships. Carbapenem resistance was consistently related to the production of oxacillinases, mostly the plasmid-mediated OXA-58 enzyme. Strains producing the OXA-23 enzyme (chromosomally mediated) were also detected. Seven PFGE clones were identified, some of which being related to international (ICL- I and ICL-II) or national clonal lineages. Multiplex PCRs identified 4 different groups (group 2 being dominant), further distinguishable in 6 sequence types by MLST. The heterogeneity of profiles highlights the diffusion of international and national clonal lineages in Italy. Continuous surveillance is needed for monitoring the spread of these worrisome strains equipped with multiple drug resistance mechanisms., (Copyright © 2013 Elsevier Inc. All rights reserved.)

Beginning on April 2007, a prospective multicenter study was performed to investigate prevalence and epidemiology of microbial pathogens causing bloodstream infections (BSIs). Twenty microbiology laboratories participated to the survey over a 1-year period. A total of 11,638 episodes of BSI occurred in 11 202 patients, with 8.5% (n=985) of episodes being polymicrobial. Of 12 781 causative organisms, aerobic Gram-negative bacteria were 47.4% (n=6058), whereas Gram-positives accounted for 43.9% (n=5608). The remaining organisms included fungal species (n=924, 7.2%) and anaerobes (n=191, 1.5%). The most prevalent agents were Escherichia coli (21.7%), Staphylococcus aureus (14.9%), Staphylococcus epidermidis (8.2%), Pseudomonas aeruginosa (7.0%), and Enterococcus faecalis (6.3%). Isolates recovered from patients admitted to medical, surgical, and intensive care units accounted for 62.9%, 17.7%, and 19.4% of cases, respectively. BSIs were classified as hospital-acquired in 67.2% of cases. Compared with previous studies, our data show an increasing role of Gram-negative bacteria among both hospital- and community-acquired blood isolates., (Copyright © 2011 Elsevier Inc. All rights reserved.)

We evaluated the ability of 60 Italian clinical microbiology laboratories in detecting and reporting beta-lactam resistance phenotypes in Enterobacteriaceae. Laboratories received 5 well-characterized isolates producing extended-spectrum beta-lactamases (ESBLs), 2 hyperproducers of chromosomal enzymes, and 3 quality control strains. The performances in antimicrobial susceptibility testing (AST) were different depending on the species and type of ESBL produced. High rates of very major errors (up to 56%) were observed for ESBL producers when testing cephalosporins and aztreonam, especially in the case of CTX-M-1-producing Escherichia coli and TEM-52-producing Proteus mirabilis. Isolates hyperproducing chromosomal enzymes were erroneously reported as ESBL producers in approximately 20% of cases. Detection of ESBLs is still a problem for clinical microbiology laboratories. Overall, performances in AST appear to be better with Klebsiella spp. producing well-known enzymes (e.g., SHV type) than with strains producing emerging enzymes (e.g., CTX-M type) or organisms not well recognized as ESBL producers (e.g., P. mirabilis).

The prevalence and the type(s) of metallo-beta-lactamases (MBLs) produced by isolates of Pseudomonas aeruginosa were investigated. During 2001, 506 nonduplicate isolates were obtained from hospitalized patients. Eighty-two strains were selected because of resistance to carbapenems and/or ceftazidime. Screening for MBL production was performed in the latter isolates by the Etest MBL strips (AB Biodisk, Solna, Sweden) and by a broth microdilution method measuring minimum inhibitory concentrations (MICs) of imipenem alone and in the presence of metal-chelating agents (EDTA and o-phenanthroline). Specific DNA probes were used to investigate the presence of genes coding for IMP- or VIM-type enzymes. Overall, four isolates of P. aeruginosa (obtained from independent patients) were found to carry a blaVIM gene. Polymerase chain reaction (PCR) experiments and DNA sequencing revealed that the VIM-2 determinant was present in three cases, whereas VIM-1 was detected in one isolate. Surveillance programs should be adopted to avoid the spread of these worrisome resistance genes.