Staphylococcal cassette chromosome mec typing and meca sequencing in methicillin-resistant staphylococci from Algeria: A highly diversified element with new mutations in mecA

Genetic mechanisms of methicillin resistance are still relevant in staphylococci. The aims of this study are to assess the possible exchanges of staphylococcal cassette chromosome mec (SCCmec) among isolates of methicillin-resistant staphylococci (MRS) and to check for known or new mutations in mecA DNA. A total of 35 MRS non-repetitive isolates were recovered, including 20 Staphylococcus haemolyticus, 7 Staphylococcus aureus, 4 Staphylococcus sciuri, 2 Staphylococcus saprophyticus and 1 isolate each of Staphylococcus xylosus and Staphylococcus lentus. Only 16 of the 35 strains were assigned to known SCCmec types: 7 SCCmec VII, 6 SCCmec IV and 3 SCCmec III, with possible horizontal transfer of the SCCmec VII from methicillin-resistant S. haemolyticus to methicillin-susceptible S. aureus. mecA gene sequencing in ten selected isolates allowed description of nine punctual mutations, seven of which were reported for the first time. The most frequent mutation was G246E, identified in isolates of methicillin-resistant S. aureus, S. sciuri, S. saprophyticus and S. lentus. These results emphasized the high degree of genetic diversity of SCCmec element in MRS and describe new missense mutations in mecA, which might be important in understanding the evolution of methicillin and new b-lactam resistance

Enhanced surveillance of invasive listeriosis in the Lombardy region, Italy, in the years 2006-2010 reveals major clones and an increase in serotype 1/2a.

Background Invasive listeriosis is a rare, life-threatening foodborne disease. Lombardy, an Italian region accounting for 16% of the total population, reported 55% of all listeriosis cases in the years 2006-2010. The aim of our study was to provide a snapshot of listeriosis epidemiology in this region after the implementation of a voluntary laboratory-based surveillance system. Methods We characterized by serotyping, pulsed-field gel electrophoresis, multilocus sequence typing and detection of epidemic clone markers, 134 isolates from 132 listeriosis cases, including 15 pregnancy-related cases, occurring in the years 2006-2010 in Lombardy. Demographic and clinical characteristics of cases have also been described. Results The mean age of non pregnancy-associated cases was 64.7\u2009years, with 55.9% of cases being older than 65\u2009years. Cases having no underlying medical conditions accounted for 11.6%. The all-cause fatality rate of 83 cases with a known survival outcome was 25.3%. Serotypes 1/2a and 4b comprised 52.2% and 38.8% of isolates, respectively. Seventy-three AscI pulsotypes and 25 sequence types assigned to 23 clonal complexes were recognized. Moreover, 53 (39.5%) isolates tested positive for the epidemic clone markers. Twelve molecular subtype clusters including at least three isolates were detected, with cluster 11 (1/2a/ST38) including 31 isolates identified during the entire study period. No outbreaks were notified to public health authorities during this period. Conclusions The findings of our study proved that epidemiology of listeriosis in Lombardy is characterized by a high prevalence of major clones and the increasing role of serotype 1/2a. Molecular subtyping is an essential tool in the epidemiology and surveillance of listeriosis. Rapid molecular cluster detection could alert about putative outbreaks, thus increasing the chance of detecting and inactivating routes of transmission

Typing methods used in the molecular epidemiology of microbial pathogens: a how-to guide

Microbial typing is often employed to determine the source and routes of infections, confirm or rule out outbreaks, trace cross-transmission of healthcare-associated pathogens, recognize virulent strains and evaluate the effectiveness of control measures. Conventional microbial typing methods have occasionally been useful in describing the epidemiology of infectious diseases. However, these methods are generally considered too variable, labour intensive and time-consuming to be of practical value in epidemiological investigations. Moreover, these approaches have proved to be insufficiently discriminatory and poorly reproducible. DNA-based typing methods rely on the analysis of the genetic material of a microorganism. In recent years, several methods have been introduced and developed for investigation of the molecular epidemiology of microbial pathogens. Each of them has advantages and limitations that make them useful in some studies and restrictive in others. The choice of a molecular typing method therefore will depend on the skill level and resources of the laboratory and the aim and scale of the investigation. This study reviews the most popular DNA-based molecular typing methods used in the epidemiology of bacterial pathogens together with their advantages and limitations

Knowledge, attitudes and self-reported practices of food service staff in nursing homes and long-term care facilities

The aim of this study was to investigate knowledge, attitudes and practices of food service staff in nursing homes and long-term care facilities for the elderly in Sicily, Italy. Association with some demographic and work-related variables was also investigated. This survey provides information and outlines many complex questions concerning the basics of food hygiene. Education level, length of service in the employment and attending courses on food hygiene influenced the knowledge, attitudes and practices of food service staff. This study has evidenced the need for continuous training among food service staff regarding food safety in LTCF and nursing homes

OXA-163-producing Klebsiella pneumoniae in Cairo, Egypt, in 2009 and 2010.

Two genetically unrelated OXA-163-carrying Klebsiella pneumoniae strains were identified from two infection cases in June 2009 and May 2010 in Cairo, Egypt. OXA-163-producing Enterobacteriaceae had been previously reported in Argentina only. Both patients had no history of travel abroad. The emergence of this newly recognized OXA-48-related \u3b2-lactamase able to hydrolyze cephalosporins and carbapenems is especially worrying in a geographic area where OXA-48 is endemic and effective surveillance for antibiotic resistance is largely unaffordable

Descriptive Epidemiology of Nasal Carriage of Staphylococcus aureus and Methicillin-Resistant Staphylococcus aureus Among Patients Admitted to Two Healthcare Facilities in Algeria

Aim: To evaluate nasal carriage rate and variables associated with Staphylococcus aureus and methicillin-resistant Staphylococcus aureus (MRSA) in patients admitted in two healthcare facilities. Results: S. aureus was isolated from 159 (26%) of the enrolled patients. Methicillin-susceptible S. aureus was isolated from 150 (24.5%) patients, and MRSA was isolated from 9 (1.5%). Cancer and previous hospitalization were associated with a significantly higher frequency of nasal S. aureus carriage among the patients admitted to the general hospital and the nephrology department, respectively. MRSA isolates were heterogeneous with respect to their staphylococcal cassette chromosome mec element (SCCmec) type, sequence type (ST), and toxin genes (pvl and tst1) content. Four isolates were attributed with the ST80-MRSA-IV clone, which is known to be predominant in Algeria. Conclusions: This is the first assessment of S. aureus and MRSA nasal carriage and associated variables in Algeria. Our findings provide also a picture of the MRSA strains circulating in the community in this geographic area. They can be useful as a guide for implementing screening and control procedures against S. aureus/MRSA in the Algerian healthcare facilities

Knowledge, risk perception and behaviours in swimming pool users of Palermo city, Sicily

To assess knowledge, risk perception and behaviours of a sample of swimming pool users in Palermo. A total of 498 subjects were interviewed by self-administered anonymous questionnaire including socio-demographic questions, knowledge/risk perception and behaviours. Overall, 289 subjects (58%) had a mean of 1.9\ub11.2 healthy behaviours. Age older than 30 years (OR=0.46; 95% CI=0.26\u20130.81), female sex (OR=0.52; 95% CI=0.35\u20130.76) and a high knowledge score (OR=0.15; 95% CI=0.03\u20130.69) were significantly associated with having healthy behaviours in both univariate and multivariate analysis. Unhealthy behaviours can be frequently found in swimming pool bathers and, particularly, in some target groups. Public health prevention strategies should be implemented to improve public awareness and information on a safe use of recreational water environments

Surveillance of enteric virus infections in a neonatal intensive care unit.

Objective. To investigate the epidemiology of neonatal viral gastroenteritis compared to the circulation of enteric viruses in children, 109 newborns in the NICU of Mother and Child Department and 214 children with enteritis admitted to the \u201cG. Di Cristina\u201d Children\u2019s Hospital in Palermo were monitored for Rotavirus, Adenovirus, Astrovirus and Norovirus infections. Methods. Stool samples were examined by EIA to detect viral antigens. Rotavirus strains were subjected to P- and G-typing. Results. A Norovirus strain was detected in one neonatal stool specimen whereas an astrovirus strain was dectected in two neonatal specimens. No Rotavirus or Adenovirus infection was identified among the newborn infants, while Rotavirus infections were detected in 24.8% of the symptomatic children. Type G4P[8] constituted 43.4% of the Rotavirus strains, followed by G2P[4] (18.9%), G3P[8] (17%), G1P[8] (13.2%) and G9P[8] (1.9%). Overall, Norovirus, Adenovirus and Astrovirus strains were responsible for 15.4% of infections in the paediatric population with diarrhoea. Conclusions. Viruses are diffuse agents of infection in children with enteritis. Virological tests have to be performed to diagnose enteric infections in the paediatric population. Maternal immunity to common Rotavirus strains combined with the limited circulation of the emerging G9 Rotavirus type among our population may account for the absence of Rotavirus infections in newborn infants

Double copies of blaKPC-3::Tn4401a on an IncX3 plasmid in Klebsiella pneumoniae successful clone ST512 from Italy

A carbapenem-resistant sequence type 512 (ST512) Klebsiella pneumoniae carbapenemase 3 (KPC-3)-producing K. pneumoniae strain showing a novel variant plasmid content was isolated in Palermo, Italy, in 2014. ST512 is a worldwide successful clone associated with the spread of blaKPC genes located on the IncFIIk pKpQIL plasmid. In our ST512 strain, the blaKPC-3 gene was unusually located on an IncX3 plasmid, whose complete sequence was determined. Two copies of blaKPC-3::Tn4401a caused by intramolecular transposition events were detected in the plasmid

Successful control of an outbreak of colonization by Klebsiella pneumoniae carbapenemase-producing K.\ua0pneumoniae sequence type 258 in a neonatal intensive care unit, Italy

This article reports an outbreak of colonization by Klebsiella pneumoniae carbapenemase-producing K. pneumoniae (KPC-Kp) sequence type (ST) 258 in a neonatal intensive care unit (NICU) in Palermo, Italy. KPC-Kp ST258 was detected by an active surveillance culture programme. Between 18th September and 14th November 2012, KPC-Kp was isolated from 10 out of 54 neonates admitted in the outbreak period. No cases of infection were recorded. Male sex was associated with colonization, whereas administration of ampicilline-sulbactam plus gentamicin was protective. Infection control interventions interrupted the spread of KPC-Kp without the need to close the NICU to new admissions. (C) 2013 The Healthcare Infection Society. Published by Elsevier Ltd. All rights reserved