Highly epidemic strains of methicillin-resistant Staphylococcus aureus (MRSA) do not differ from other MRSA or methicillin-sensitive strains in capsule formation, Protein A content or adherence to HEp-2 cells

During the 1990s, two strains of epidemic methicillin-resistant Staphylococcus aureus, designated ‘phage types EMRSA-15 and EMRSA-16, have emerged as significant hospital pathogens. They have resisted standard methods of control and spread widely amongst in the UK, often becoming endemic, while the incidence of other epidemic types of MRSA has either declined or not changed. This suggests that EMRSA-15 and EMRSA-16 possess special properties that favour their dissemination and survival. In order to investigate this hypothesis, a study was undertaken that examined methicillin-sensitive and methicillinresistant strains of Staphylococcus aureus, including EMRSA types 1, 2, 3, 15 and 16, for capsule formation, the amount of bound protein A produced, and quantitative adherence to the human continuous epithelial cell line HEp-2. Although all these properties varied amongst the strains examined, there was no relationship between any of them and methicillin resistance or epidemic type, and, incidentally, no relationship between cell-wall bound protein A content and adherence

Device-independent, real-time identification of bacterial pathogens with a metal oxide-based olfactory sensor

A novel olfactory method for bacterial species identification using an electronic nose device called the MonoNose was developed. Differential speciation of micro-organisms present in primary cultures of clinical samples could be performed by real-time identification of volatile organic compounds (VOCs) produced during microbial replication. Kinetic measurements show that the dynamic changes in headspace gas composition are orders of magnitude larger than the static differences at the end of fermentation. Eleven different, clinically relevant bacterial species were included in this study. For each of the species, two to eight different strains were used to take intra-species biodiversity into account. A total of 52 different strains were measured in an incubator at 37°C. The results show that the diagnostic specificities varied from 100% for Clostridium difficile to 67% for Enterobacter cloacae with an overall average of 87%. Pathogen identification with a MonoNose can be achieved within 6–8 h of inoculation of the culture broths. The diagnostic specificity can be improved by broth modification to improve the VOC production of the pathogens involved

Prevalence and role of efflux pump activity in ciprofloxacin resistance in clinical isolates of Klebsiella pneumoniae

International audienceWe investigated the prevalence and role of efflux pump activity and possible drug influx resistance in ciprofloxacin susceptibility amongst 26 distinct clinical isolates of of varying ciprofloxacin susceptibilities and known quinolone resistance-determining region (QRDR) genotypes. Cellular [C]ciprofloxacin accumulation patterns and the amount of cell-associated [C]ciprofloxacin of mid-logarithmic phase cells were determined before and after challenging with the efflux pump inhibitor carbonyl cyanide 3-chlorophenylhydrazone (CCCP). Most isolates (24/26), and all with ciprofloxacin minimum inhibitory concentrations (MICs) >1 µg/ml, had efflux activity that could extrude up to 90% of cell-associated [C]ciprofloxacin; none had significant influx resistance. In isolates with no QRDR mutations, efflux alone reduced ciprofloxacin susceptibility. In isolates with QRDR mutations, the efflux activity varied: in one isolate with no efflux activity, the most common fluoroquinolone resistance-causing QRDR mutation did not bring about clinically significant ciprofloxacin resistance; isolates with multiple mutations had high MICs and, usually, high levels of efflux activity. Fluoroquinolone efflux activity is much more common in clinical isolates of than previously reported and it can contribute to decreased ciprofloxacin susceptibility

Hypermutability in clinical isolates of Klebsiella pneumoniae is uncommon and is unrelated to ciprofloxacin resistance

International audienceWe investigated hypermutability in and its association with ciprofloxacin resistance and mutations in the quinolone resistance-determining region (QRDR). Sixty-four strains of isolated in London, UK, between 1995 and 2002 with widely differing ciprofloxacin minimum inhibitory concentrations (MICs) and known and sequences were tested for mutation frequencies by selection with rifampicin. Only three hypermutable (frequency ≥10) strains were identified, with ciprofloxacin MICs of 0.25, 8 and 64μg/mL. There was no relationship between hypermutation and the ciprofloxacin MIC or QRDR mutations. Screening selected strains with streptomycin did not reveal any hypermutators, and screening with ciprofloxacin identified only two of the three hypermutators identified by rifampicin. Hypermutation in is uncommon and does not contribute to accumulation of QRDR mutations or directly to ciprofloxacin resistance

A novel plasmid pIJB1 possessing a putative 2,4-dichlorophenoxyacetate degradative transposon Tn5530 in Burkholderia cepacia strain 2a

A 102-kb plasmid, pIJB1, was isolated from Burkholderia cepacia strain 2a, which is able to use 2,4-dichlorophenoxyacetate (2,4-D) as a sole carbon source, and a physical map of the plasmid has been established. It was observed that spontaneous loss of a 37-kb fragment of the plasmid after growth in nonselective medium occurred, generating a plasmid of diminished size, pIJB2. The deletion event is concomitant with the loss of the 2,4-D dissimilatory phenotype, indicating that at least some of the 2,4-D degradative genes are on the missing fragment. The missing fragment is flanked by two identical 4.3-kb insertion sequences (IS) and shows a typical composite transposon structure of 41-kb in size, designated Tn5530. The mutant plasmid pIJB2 possesses a single copy of the IS element