Virulence and resistance pattern of a novel sequence type of linezolid-resistant Enterococcus faecium identified by whole-genome sequencing.

Empirical use of linezolid has been advocated in neutropenic febrile patients colonised by vancomycin-resistant enterococci (VRE) because of the risk of bloodstream infection (BSI). This study aimed to genetically describe a vancomycin-resistant Enterococcus faecium (VREfm) BSI isolate resistant to linezolid (VRLRE) in a patient previously colonised by VREfm and to determine the incidence of colonisation and infection by VREfm in a bone marrow transplant unit over a 10-year period. Data for VREfm colonisation and infection were evaluated. PCR for the vanA and vanB genes, pulsed-field gel electrophoresis (PFGE) and microdilution antimicrobial susceptibility testing (vancomycin, teicoplanin, linezolid and aminoglycosides) were performed. Three isolates, including the VRLRE, were selected for whole-genome sequencing by Ion Torrent™, with E. faecium CP006620-Aus0085 used as a reference. Eighty-seven VREfm were analysed; all were linezolid-susceptible and harboured vanA, except for one blood isolate from a febrile neutropenic patient colonised by VREfm who received linezolid for 12 days and developed a BSI by VRLRE (linezolid MIC≥8µg/mL). Linezolid resistance was associated with a G2576T mutation in the 23SrRNA gene. PFGE analysis demonstrated that the 87 isolates belonged to four major clusters; however, the VRLRE presented only 50% similarity. Three sequence types (STs) were identified: ST412 (the predominant clone, which was more virulent compared with the other isolates); ST478 (linezolid-susceptible VREfm); and a novel ST named ST987 (VRLRE). SNP analysis showed a higher similarity between linezolid-susceptible VREfm and the predominant clone compared with VRLRE. VRLRE presented a G2576T mutation and belonged to a novel ST (ST987).

Pseudooutbreak of rapidly growing mycobacteria due to Mycobacterium abscessus subsp bolletii in a digestive and respiratory endoscopy unit caused by the same clone as that of a countrywide outbreak.

BACKGROUND: The nontuberculous mycobacteria (NTM) are widely spread. In Brazil, 2,520 cases of rapidly growing mycobacteria (RGM) infections after medical procedures were reported, with 5.4% of cases related to nonsurgical invasive procedures and with an occurrence of 1 clone (BRA100) of Mycobacterium abscessus subsp bolletii. OBJECTIVE: To describe a pseudooutbreak of M abscessus subsp bolletii in an endoscopy and bronchoscopy unit. METHODS: The alert for a pseudooutbreak was given when 3 patients, in the same week, had a positive bronchoalveolar lavage culture for M abscessus subsp bolletii. The patients had no symptoms/signs of mycobacterial infection; thus, contamination of bronchoscopes was suspected. Samples for culturing were collected from bronchoscopes, digestive endoscopes, automated disinfection machines, and the water supply. Clinical samples were identified by polymerase chain reaction restriction-enzyme analysis (PRA) of the hsp65 gene and their pulsed-field gel electrophoresis pattern was compared with environmental samples. RESULTS: The investigation demonstrated a contamination of bronchoscopes, digestive endoscopes, and disinfection machines. Molecular typing demonstrated that all strains belonged to the same clone (MAB01), identical to clone BRA100. DISCUSSION: Cross-transmission due to poor disinfection as well as resistance to glutaraldehyde may play roles in the spread of MAB01 M abscessus subsp bolletii, which may have a unique resistance to the environment and adaption to human hosts. However the water supply may have played a role. Attention is needed to ensure the quality of water used to rinse disinfected equipment.

Polyclonal outbreak of bloodstream infections caused by Burkholderia cepacia complex in hematology and bone marrow transplant outpatient units.

AIM: The objective was to describe an outbreak of bloodstream infections by Burkholderia cepacia complex (Bcc) in bone marrow transplant and hematology outpatients. METHODS: On February 15, 2008 a Bcc outbreak was suspected. 24 cases were identified. Demographic and clinical data were evaluated. Environment and healthcare workers' (HCW) hands were cultured. Species were determined and typed. Reinforcement of hand hygiene, central venous catheter (CVC) care, infusion therapy, and maintenance of laminar flow cabinet were undertaken. 16 different HCWs had cared for the CVCs. Multi-dose heparin and saline were prepared on counter common to both units. FINDINGS: 14 patients had B. multivorans (one patient had also B. cenopacia), six non-multivorans Bcc and one did not belong to Bcc. Clone A B. multivorans occurred in 12 patients (from Hematology); in 10 their CVC had been used on February 11/12. Environmental and HCW cultures were negative. All patients were treated with meropenem, and ceftazidime lock-therapy. Eight patients (30%) were hospitalized. No deaths occurred. After control measures (multidose vial for single patient; CVC lock with ceftazidime; cleaning of laminar flow cabinet; hand hygiene improvement; use of cabinet to store prepared medication), no new cases occurred. CONCLUSIONS: This polyclonal outbreak may be explained by a common source containing multiple species of Bcc, maybe the laminar flow cabinet common to both units. There may have been contamination by B. multivorans (clone A) of multi-dose vials.