Maria Bagattini

Molecular Epidemiology of Sequential Outbreaks of Acinetobacter baumannii in an Intensive Care Unit Shows the Emergence of Carbapenem Resistance

ABSTRACT The molecular epidemiology of multidrug-resistant Acinetobacter baumannii was investigated in the medical-surgical intensive care unit (ICU) of a university hospital in Italy during two window periods in which two sequential A. baumannii epidemics occurred. Genotype analysis by pulsed-field gel electrophoresis (PFGE) of A. baumannii isolates from 131 patients identified nine distinct PFGE patterns. Of these, PFGE clones B and I predominated and occurred sequentially during the two epidemics. A. baumannii epidemic clones showed a multidrug-resistant antibiotype, being clone B resistant to all antimicrobials tested except the carbapenems and clone I resistant to all antimicrobials except ampicillin-sulbactam and gentamicin. Type 1 integrons of 2.5 and 2.2 kb were amplified from the chromosomal DNA of epidemic PFGE clones B and I, respectively, but not from the chromosomal DNA of the nonepidemic clones. Nucleotide analysis of clone B integron identified four gene cassettes: aacC1, which confers resistance to gentamicin; two open reading frames (ORFs) coding for unknown products; and aadA1a, which confers resistance to spectinomycin and streptomycin. The integron of clone I contained three gene cassettes: aacA4, which confers resistance to amikacin, netilmicin, and tobramycin; an unknown ORF; and blaOXA-20, which codes for a class D β-lactamase that confers resistance to amoxicillin, ticarcillin, oxacillin, and cloxacillin. Also, the blaIMP allele was amplified from chromosomal DNA of A. baumannii strains of PFGE type I. Class 1 integrons carrying antimicrobial resistance genes and blaIMP allele in A. baumannii epidemic strains correlated with the high use rates of broad-spectrum cephalosporins, carbapenems, and aminoglycosides in the ICU during the study period.

A Plasmid-Borne blaOXA-58 Gene Confers Imipenem Resistance to Acinetobacter baumannii Isolates from a Lebanese Hospital

ABSTRACT We investigated the basis of the carbapenem resistance of 17 multidrug-resistant Acinetobacter baumannii clinical isolates collected from 2004 to 2005 at the Saint George University Hospital in Beirut, Lebanon. A. baumannii isolates were clonally related and were susceptible to colistin and trimethoprim-sulfamethoxazole, susceptible or intermediate to ampicillin-sulbactam and meropenem, and resistant to all other antimicrobials. Conjugation experiments demonstrated that resistance to imipenem could be transferred along with a plasmid containing the carbapenem-hydrolyzing oxacillinase blaOXA-58 gene. The plasmid that we called pABIR was 29,823 bp in size and showed a novel mosaic structure composed of two origins of replication, four insertion sequence (IS) elements, and 28 open reading frames. The blaOXA-58 gene was flanked by IS18 and ISAba3 elements at the 5′ and 3′ ends, respectively. The production of the carbapenem-hydrolyzing oxacillinase OXA-58 was apparently the only mechanism for carbapenem resistance in A. baumannii isolates causing the outbreak at the Lebanese Hospital.

Molecular epidemiology of carbapenem resistant Enterobacteriaceae in Valle d’Aosta region, Italy, shows the emergence of KPC-2 producing Klebsiella pneumoniae clonal complex 101 (ST101 and ST1789)

BackgroundThe spread of carbapenem resistant Enterobacteriaceae (CRE) is an emerging clinical problem, of great relevance in Europe and worldwide. The aim of this study was the molecular epidemiology of CRE isolates in Valle d’Aosta region, Italy, and the mechanism of carbapenem resistance.ResultsSixty consecutive CRE samples were isolated from 52 hospital inpatients and/or outpatients from November 2013 to August 2014. Genotyping of microbial isolates was done by pulsed-field gel electrophoresis (PFGE) and multi-locus sequence typing (MLST), carbapenemases were identified by PCR and sequencing. Carbapenem resistance gene transfer was performed by filter mating, plasmids from parental and transconjugant strains were assigned to incompatibility groups by PCR-based replicon typing. Molecular characterization of CRE isolates assigned 25 Klebsiella pneumoniae isolates to PFGE types A1-A5 and sequencing type (ST) 101, 17 K. pneumoniae isolates to PFGE type A and ST1789 (a single locus variant of ST101), 7 K. pneumoniae isolates to PFGE types B or C and ST512, 2 K. pneumoniae isolates to PFGE type D and ST405, and 5 Escherichia coli isolates to PFGE type a and ST131. All K. pneumoniae ST101 and ST1789 isolates were extended-spectrum beta-lactamase (ESBL) producers and carried blaCTX-M-1 group gene; 4 K. pneumoniae ST101 isolates were resistant to colistin. Molecular analysis of beta-lactamase genes identified blaKPC-2 and blaCTX-M-group 1 into conjugative plasmid/s assigned to IncFII incompatibility group in ST101 and ST1789 K. pneumoniae isolates, blaKPC-3 into conjugative plasmid/s assigned to IncF incompatibility group in ST512 and ST405 K. pneumoniae isolates, blaVIM-1 into conjugative plasmid/s assigned to IncN incompatibility group in ST131 E. coli isolates.ConclusionsThe spread of CRE in Valle d’Aosta region was caused by the selection of KPC-2 producing K. pneumoniae ST101 and ST1789 epidemic clones belonging to clonal complex 101, KPC-3 producing K. pneumoniae epidemic clones assigned to ST512 and ST405, and VIM-1 producing E.coli ST131 epidemic clone. Carbapenem resistance, along with blaKPC-2, blaKPC-3 and blaVIM-1 carbapenemase genes, was transferred by conjugative plasmids assigned to IncFII, IncF, and IncN incompatibility groups, respectively, in filter mating experiments. The emergence of colistin resistance was observed in KPC-2 producing K. pneumoniae ST101 isolates.

Risk factors for Candida parapsilosis bloodstream infection in a neonatal intensive care unit: a case-control study

BackgroundCandida parapsilosis is increasingly responsible for invasive candidiasis in neonates. This study investigates phenotypic and genotypic features of C. parapsilosis microbial isolates and underlying clinical conditions associated with acquisition of C. parapsilosis in a neonatal intensive care unit (NICU) in Italy.MethodsIdentification of C. parapsilosis was performed by VITEK® 2 and MALDI TOF and confirmed by analysis of internal transcribed spacer ribosomal DNA sequences. Genotyping was performed by PCR fingerprinting. Antifungal susceptibility of strains was evaluated by microdilution. A case-control study was designed to identify risk factors for C. parapsilosis bloodstream infection.ResultsDuring the study period (April 2009- April 2012), C. parapsilosis was responsible for 6 umbilical catheter and 11 central catheter-associated bloodstream infection in 17 neonates in the NICU. Molecular typing identified identical fingerprinting profile in all C. parapsilosis isolates from neonates. Fifteen of 17 C. parapsilosis isolates were susceptible to all antifungal drugs, two isolates were resistant to fluconazole and intermediate susceptible to itraconazole. Low birthweight, gestational age and time to exposure to assisted ventilation were risk factors for C. parapsilosis infection in neonates in the NICU at univariate and multivariate analysis.ConclusionC. parapsilosis bloodstream infections in the NICU were caused by a single epidemic clone. Low birthweight, gestational age and time to exposure to invasive devices, with predominance of assisted ventilation, were the clinical conditions associated with C. parapsilosis bloodstream infection in the NICU.

Microbiological evaluation of the efficacy of two new biodetergents on multidrug-resistant nosocomial pathogens

BackgroundIn the last few years, several outbreaks of nosocomial infections caused by multidrug-resistant pathogenic agents have been observed, and various biocides products were developed in order to control this phenomenon. We investigated the efficacy of two natural biodetergents composed of plants and kelps extracts, BATT1 and BATT2, against multidrug-resistant strains.MethodsIn-vitro antibacterial efficacy of BATT1 and BATT2 against nosocomial multidrug-resistant isolates was assessed using a suspension-inhibition test, with and without bovine serum albumin (BSA). The test was also carried out on glass surfaces with and without BSA.ResultsIn vitro tests with both biocidal disinfectants at 25% concentration demonstrated an overall drop in bacterial, mould and yeast counts after 10 min of contact with or without organic substances. For Pseudomonas aeruginosa, it was necessary to use undiluted disinfectants with and without an organic substance. The same results were obtained in tests carried out on glass surfaces for all strains.ConclusionsThe natural products BATT1 and BATT2 behave like good biocides even in presence of organic substances. The use of both disinfectants may be beneficial for reducing hospital-acquired pathogens that are not susceptible to disinfectants.However, it has to be stressed that all these experiments were carried out in vitro and they still require validation from use in clinical practice.

Automated cleaning of fan coil units with a natural detergent-disinfectant product

BackgroundAir conditioning systems represent one important source of microbial pollutants for indoor air. In the past few years, numerous strategies have been conceived to reduce the contamination of air conditioners, mainly in hospital settings. The biocidal detergent BATT2 represents a natural product obtained through extraction from brown seaweeds, that has been tested previously on multidrug-resistant microorganisms.MethodsBATT2 has been utilized for the disinfection of fan coil units from four air conditioning systems located in hospital environments with a mean degree of risk. Samples were collected from the air supplied by the conditioning systems and from the surfaces of fan coil units, before and after sanitization procedures. Total microbial counts at 37°C and 22°C and mycotic count at 32°C were evaluated. Staphylococci and Pseudomonas aeruginosa were also detected on surfaces samples.ResultsThe biodetergent was able to reduce up 50% of the microbial pollution of fan coil units surfaces and air supplied by the air conditioners.ConclusionsBATT2 could be considered for cleaning/disinfection of air conditioning systems, that should be performed on the basis of accurate and verifiable sanitization protocols.

Acinetobacter Infections in Neonates

Purpose of ReviewMDR-Gram-negative bacteria are a great concern in the neonatal population, with a worldwide rise in the reported incidence and with very limited therapeutic options. Acinetobacter baumannii is responsible for many infections in neonates and outbreaks in neonatal intensive care unit (NICU); also, outbreaks caused by other Acinetobacter species have been reported. The aim of this review is to document the epidemiology of Acinetobacter spp. infections in neonates and risk factors for acquisition of Acinetobacter spp. in the NICU using data from published studies.Recent FindingsAcinetobacter spp. infections are increasing in neonates in NICU. Outbreak caused by multidrug resistant (MDR) or extensively drug resistant (XDR) A. baumannii but also outbreak caused by susceptible A. soli and A. septicus sp. nov., were reported in neonates. Acinetobacter spp. were responsible for bloodstream infections and respiratory tract infections in neonates. Risk factors for A. baumannii acquisition in neonates were low birthweight, length of NICU stay, umbilical catheterization, central-venous catheterization, assisted ventilation, and prior antibiotic use.SummaryThis review highlights the importance of surveillance of risk factors for healthcare-associated infections in NICU to control MDR and XDR A. baumannii infections in neonates.