Colonization of residents and staff of an Italian long-term care facility and an adjacent acute care hospital geriatric unit by multidrug-resistant bacteria.

In 2016, we undertook a point prevalence screening study for Enterobacteriaceae with extended-spectrum ß-lactamases (ESBLs), high-level AmpC cephalosporinases and carbapenemases, and also methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin resistant enterococci (VRE) in a long-term care facility (LTCF) and the associated acute care hospital geriatric unit in Bolzano, Northern Italy. Urine samples and rectal, inguinal, oropharyngeal and nasal swabs were plated on selective agars. Demographic data were collected. ESBL and carbapenemase genes were sought by PCR. We found the following colonization percentages with multidrug-resistant (MDR) bacteria in 2016 in LTCF residents: all MDR organisms, 66.1%; ESBL producers, 53.0%; carbapenemase-producers, 1.7%; MRSA, 14.8%; VRE, 0.8%. Colonization by all MDR bacteria was 19.4% for LTCF staff and 26.0% for geriatric unit patients. PCR showed that 80.3% of Escherichia coli isolates from LTCF residents, all E. coli isolates from LTCF staff, 62.5% and 100% of Klebsiella pneumoniae from LTCF residents and geriatric unit patients, respectively, had a blaCTX-M-type gene. All carbapenemase-producing Enterobacteriaceae harboured a blaVIM-type gene. To conclude, the ongoing widespread diffusion of MDR bacteria in the LTCF suggests that efforts should be strengthened on MDR screening, implementation of infection control strategies and antibiotic stewardship programs targeting the unique aspects of LTCFs.

Carbapenemase-producing Enterobacteriaceae during 2011-12 in the Bolzano area (Northern Italy): increasing diversity in a low-endemicity setting.

The recent (2011-2012) distribution of carbapenemase determinants in Enterobacteriaceae was studied in the Bolzano area (Northern Italy). Low proportions of carbapenemase producers were found for Escherichia coli (0.2%), Citrobacter freundii (1.1%), Klebsiella pneumoniae (1.3%), Klebsiella oxytoca (1.6%) and Enterobacter spp (1.8%). Although VIM-1 remained the most common carbapenemase, the emergence of K. pneumoniae producing KPC-3 and of E. coli producing OXA-48 was observed. Of concern is the spread of the hyperepidemic strains E. coli ST131 producing VIM-1 and K. pneumoniae ST258 producing KPC-3. Low essential and category agreements between the reference broth microdilution and commercial methods were observed for carbapenems.

Evaluation of a "home-made" method to determine viral genotype and characterize mutations conferring drug resistance in chronic hepatitis B patients.

We compared a home-made sequencing system to analyze plasma samples from patients with chronic HBV infection with the commercial TRUGENE(®) HBV Genotyping Assay. A PCR and sequencing protocol based on published primers was applied to detect the viral genotypes as well as the major patterns of point mutations leading to resistance to lamivudine, adefovir and entecavir. For the determination of HBV genotypes the obtained sequences were aligned with a database created within the RIDOM TraceEdit program and publicly available reference sequences. Our results showed perfect correlation with the commercial system, with types D (72%) and A (22%) being the most frequent genotypes. The resistance loci were also reliably detected with mostly combined L180M and M204V/I mutations as the local patterns. M204I mutations were more frequent in genotype D, M204V in genotype A isolates. G173L mutations were not found. The only genotype C isolate tested revealed a different pattern (E263D and I269L). These data speak for the usability of this rapid amplification and sequencing approach for routine genotyping of HBV isolates and simultaneous determination of the drug resistance profile of the dominant viral species.

Generalized parapoxvirus infection associated with increased antibody titres for varicella zoster virus and measles.

Human parapoxvirus infections are rare, self-limiting, zoonotic diseases. A 35-year-old veterinarian presented with a generalized rash of large umbilicated vesicles that appeared after antibiotic treatment for erysipelas on the forearm. The erysipelas arose from an erupted pustular thumb lesion that appeared after examining a sheep. An outbreak of chickenpox in the village suggested parapoxvirus or varicella zoster virus (VZV) was the most likely agent. No poxvirus was detected by electron microscopy or in cell cultures from lesion material. PCR revealed parapoxvirus DNA with a sequence similar to orf-viruses from Finland. Orf-virus immunofluorescence showed a titre increase, supporting the parapoxvirus diagnosis. VZV was not detected by PCR, but varicella antibodies increased three-fold in serum samples drawn two weeks apart. In addition, the patient had high antibody titres for measles and reported recent contact with individuals exposed to an outbreak of measles in nearby Austria. To explain the unusually generalized symptoms in this young and healthy patient, these findings could be variously interpreted as: i) a booster by community VZV infections; ii) a subclinical VZV (re)infection that was superinfected by the parapoxvirus; iii) an orf-virus mediated immune stimulation; iv) a post-infectious syndrome; or v) a temporary immunosuppression by subclinical measles.