PurposeCarbapenemases-producing Klebsiella pneumoniae are challenging antimicrobial therapy of hospitalised patients, which is further complicated by colistin resistance. This study describes molecular epidemiological insights into colistin-resistant and carbapenemases-producing clinical K. pneumoniae.Patients and methodsCultures collected from 26 hospitalised patients during 2014–2017 in the main hospital in Molise Region, central Italy, were characterized. The minimum inhibitory concentration for 19 antibiotics was determined, including carbapenems and colistin. Prevalence of resistance-associated genes was investigated through PCR, detecting blaKPC, blaGES, blaVIM, blaIMP, blaNDM, blaOXA-48, blaCTX-M, blaTEM, blaSHV, and mcr-1,2,3,4,5,6,7,8. The mgrB gene was also analysed in colistin-resistant strains by PCR and sequencing assays. K. pneumoniae were typed by pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST).ResultsTwenty out of 26 K. pneumoniae were phenotypically resistant to carbapenems and 19 were resistant to colistin. All isolates harbored blaKPC, and blaSHV, blaTEM and blaVIM were further the most common resistance-associated genes. In colistin-resistant strains, mcr-1,2,3,4,5,6,7,8 variants were not detected, while mutations and insertion elements in mgrB were observed in 68.4% (n=13) in 31.6% (n=6) isolates, respectively. PFGE revealed 12 clusters and 18 pulsotypes at 85% and 95% cut-off, while the Sequence Types ST512 (n=13, 50%), ST101 (n=10, 38.5%), ST307 (n=2, 7.7%) plus a novel ST were detected using MLST.ConclusionAll K. pneumoniae showed a multidrug-resistant phenotype, particularly to carbapenems and colistin. According to national data, blaKPC was the prevailing carbapenemase, followed by blaVIM, while blaTEM and blaSHV were among the most frequent beta-lactamases. Consistent with previous reports in Italy, ST512 was the most common clone, particularly during 2014–15, whilst ST101 became dominant in 2016–17. Colistin resistance was mainly associated with deleterious mutations and transposon in the mgrB gene. Improvements of surveillance, compliance with infection prevention procedures and antimicrobial stewardship are essential to limit the spread of resistant K. pneumoniae.
Antimicrobial-resistant Klebsiella pneumoniae represent a global public health concern. K. pneumoniae strains isolated during 2010 and 2014-2016 within a single hospital of Molise Region, Central Italy, were analyzed testing antimicrobial susceptibility, clonality by pulsed-field gel electrophoresis (PFGE) and random amplified polymorphic DNA (RAPD)-PCR, and prevalence of carbapenem resistance genes by PCR. Forty isolates (23 wild-type in 2010 and 17 non-wild-type in 2014-2016) were collected from hospitalized patients (65.2 ± 18.1 years old, 75% male, 80% from intensive care unit-ICU). K. pneumoniae showed multidrug-resistant profiles and 15 resistotypes were identified (discriminatory power D = 0.88). The 69.6 and 17.4% of isolates in 2010 resulted intermediate and resistant to imipenem, respectively, and 91.3% was sensitive to meropenem, while 88.2% of isolates of 2014-2016 were resistant to both antibiotics. PFGE identified 16 clusters versus 23 by RAPD, 26 pulsotypes versus 33 RAPD patterns (D ≥ 0.97). PFGE separated strains according to isolation period and identified an outbreak occurred in the ICU during December 2014 and January 2015. No strains harbored bla, bla, bla, and bla genes, as well as AmpC plasmid-mediated beta-lactamases genes. Only K. pneumoniae isolated during 2014-2016 were bla positive. Prevalence of bla was 87 and 76.5% during 2010 and 2014-2016, respectively. No strains colistin-resistant harbored mcr-1 plasmid-mediated resistance gene. The study findings underline an increased circulation of multidrug-resistant K. pneumoniae within the hospital, and the acquisition of carbapenem resistance mechanism. The implementation of surveillance and molecular characterization of isolates are needed to identify outbreaks, reduce the spread of resistance, and guide empirical therapy.
Italy was one of the most affected nations by coronavirus disease 2019 outside China. The infections, initially limited to Northern Italy, spread to all other Italian regions. This study aims to provide a snapshot of severe acute respiratory syndrome coronavirus‐2 (SARS‐CoV‐2) epidemiology based on a single‐center laboratory experience in Rome. The study retrospectively included 6565 subjects tested for SARS‐CoV‐2 at the Laboratory of Virology of Sapienza University Hospital in Rome from 6 March to 4 May. A total of 9995 clinical specimens were analyzed, including nasopharyngeal swabs, bronchoalveolar lavage fluids, gargle lavages, stools, pleural fluids, and cerebrospinal fluids. Positivity to SARS‐CoV‐2 was detected in 8% (527/6565) of individuals, increased with age, and was higher in male patients ( P < .001). The number of new confirmed cases reached a peak on 18 March and then decreased. The virus was detected in respiratory samples, in stool and in pleural fluids, while none of gargle lavage or cerebrospinal fluid samples gave a positive result. This analysis allowed to gather comprehensive information on SARS‐CoV‐2 epidemiology in our area, highlighting positivity variations over time and in different sex and age group and the need for a continuous surveillance of the infection, mostly because the pandemic evolution remains unknown.
Background: Hand hygiene is considered as the most important strategy for preventing healthcare infections. Objective: In this single-arm study, skin tolerance, effectiveness and prolonged efficacy, and adherence to use an alcohol-based hand rub (ABHR) solution among hospital nurses was evaluated. Methods: Nurses were recruited in the main hospital of Molise Region, Central Italy. Skin reactions during 12-week follow-up were self-reported by nurses and, when occurring examined by hospital dermatologist. Samples were collected from palms and fingertips before and after ABHR, also at random times during work. Results: Dermatological reactions were not reported by 20 nurses (15 female and 5 male, aged 30–59 years) after product use. Microbial counts decreased by 99% (2Log10) in 75%, with higher reduction in palm than fingertips (99% vs 70% respectively). Analysis of six randomly collected samples after 10–20 and 30–40 min from the last use showed a satisfactory prolonged efficacy. Discussion: Beyond high effectiveness to reduce transient microbiota, no skin reactions were observed, likely due to the camomile, thyme and eucalyptus extracts contained within the gel. This study, addressing technical questions of a commercialised product, provides useful information for public health authorities faced with a choice of hand disinfectants, evaluating cost-effectiveness and cost-benefit in the light of the huge amount of these products needed at hospital level.
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