Identification and characterization of CTX-M-15 producing Klebsiella pneumoniae clone ST101 in a Hungarian university teaching hospital

We investigated the molecular epidemiology of extended spectrum β-lactamase (ESBL) producing Klebsiella pneumoniae isolates derived from the teaching hospitals of University of Pécs, Pécs, Hungary in the time period 2004–2008. Molecular typing, antimicrobial susceptibility testing, detection of common β-lactamase genes (blaCTX-M, blaTEM and blaSHV) and virulence associated traits (hypermucoviscosity, magA, k2a, rmpA, siderophores, type 1 and 3 fimbria, biofilm formation, serum resistance) were performed for 102 isolates. The results showed the presence of three major ciprofloxacin resistant CTX-M-15 producing clones (ST15 n = 69, ST101 n = 10, and ST147 n = 9), of which ST15 was predominant and universally widespread. Considering distribution in time and place, ST101 and ST147 were detected at fewer inpatient units and within a narrower time frame, as compared to ST15. Beside major clones, eleven minor clones were identified, and were shown to harbour the following β-lactamase genes: six clones carried blaCTX-M, four clones harboured blaSHV-5 and one clone possessed both blaCTX-M and ESBL type blaSHV. Among the SHV-5 producing K. pneumoniae clones a novel sequence type was found, namely ST1193, which harboured a unique infB allele. Different virulence factor content and peculiar antimicrobial susceptibility profile were characteristic for each clone. In contrast to major clone isolates, which showed high level resistance to ciprofloxacin, minor clone isolates displayed significantly lower MIC values for ciprofloxacin suggesting a role for fluoroquinolones in the dissemination of the major K. pneumoniae clones. This is the first description of the CTX-M-15 producing K. pneumoniae clone ST101 in Hungary

Genome sequences of three turkey orthoreovirus strains isolated in Hungary

We have investigated the genomic properties of three turkey reovirus strains—19831M09, D1246, and D1104—isolated in Hungary in 2009. Sequence identity values and phylogenetic calculations indicated genetic conservativeness among the studied Hungarian strains and a close relationship with strains isolated in the United States

MALDI-TOF MS versus 16S rRNA sequencing: Minor discrepancy between tools in identification of Bacteroides isolates

Members of the genus Bacteroides are important components of the normal microbiota of gastrointestinal tract; however, as opportunistic pathogens are also associated with severe or even life-threatening infections with significant mortality. Various species within Bacteroides fragilis group are phenotypically very similar; thus, their identifications with traditional-automated biochemical methods are frequently inaccurate. The identification of the newly discovered or reclassified bacteria can be doubtful because of the lack of biochemical profile in the database of these tests. The aim of this study was to determine the accuracy of matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) method by testing of 400 Hungarian Bacteroides clinical isolates. Inaccurate identification results with MALDI-TOF MS were confirmed by 16S rRNA gene sequencing and findings were compared with traditional-automated biochemical test rapid ID 32A method as well

A multicentre survey of the antibiotic susceptibility of clinical Bacteroides species from Hungary.

BACKGROUND: The species of the Bacteroides fragilis group are important components of human microbiota, but as opportunistic pathogens they can be the causative agents of severe infections. METHODS: The major aims of our investigation were the evaluation of the susceptibility of 400 different Hungarian B. fragilis group isolates to 10 antibiotics by the agar dilution method, the comparison of our resistance data with previous national and international antibiotic resistance data and the comparison of present data in regional aspect. The MIC-values on 10 antibiotics of all the strains were determined with the agar dilution method by CLSI. The presence of the cfiA gene in Division II B. fragilis strains was confirmed by RT-PCR. RESULTS: We detected a relatively high resistance rate of ampicillin, moxifloxacin, clindamycin and tetracycline, but amoxicillin/clavulanic acid, metronidazole, tigecycline and chloramphenicol showed excellent activity. In this study, we found that 6.75% of the isolates were resistant to cefoxitin and 7% to meropenem, while 8.58% of our B. fragilis strains harboured the cfiA gene. Most of the meropenem resistant strains were isolated in one of the participating centres. In the case of meropenem, cefoxitin, clindamycin and high-level-ampicillin-resistant strains, we found significant regional differences. DISCUSSION: Most of the results of our study were concordant with previous national and international data, with the exception of amoxicillin/clavulanic acid, cefoxitin and meropenem. CONCLUSIONS: Our study highlighted the importance of the periodic monitoring of the antimicrobial susceptibility of Bacteroides species providing important information for the appropriate therapy

Detection of enterotoxin and protease genes among Hungarian clinical Bacteroides fragilis isolates.

Bacteroides fragilis as a commensal bacterium is a member of the human intestinal flora, but as an opportunistic pathogen it can cause serious infections as well. Some of them, harbouring an enterotoxin gene (bft), may cause diarrhoea mainly in young children. Recently it has been shown that a member of C11 proteases called fragipain (fpn) can activate the enterotoxin, while C10 protease (bfp) is suspected of playing an important role in the invasiveness of the B. fragilis isolates. The objective of this study was to investigate the prevalence and distribution of the bft isotypes in 200 Hungarian B. fragilis isolates collected recently; and in a subset of 72 strains, we wanted to determine the prevalence of bfp1-4 and fpn genes in bft-positive and bft-negative strains. Using the MALDI-TOF MS cfiA identification project file, 19 B. fragilis strains belonging to Division II were identified and the presence of the cfiA gene was confirmed by RT-PCR. Twenty two (13.0%) B. fragilis isolates turned out to be bft gene positive by RT-PCR; 20 isolates harboured bft-1 and six bft-2 isotypes, but no bft-3 isotype containing strains were found. A melting curve analysis and the PCR-RFLP were performed to differentiate between the bft-1 and bft-2 isotypes confirmed by sequencing. Thirty eight strains harboured bfp1, 58 isolates contained bfp2 gene, while 17 isolates proved positive for bfp3. Morever, no bfp4 positive isolate was found, and some of the B. fragilis strains tested harboured two or three bfp isotypes simultaneously. Among the 26 bft-positive strains, 24 contained the fpn gene, which confirms the role of fragipain in the activation of B. fragilis enterotoxin. In experiments, a significant negative correlation between fpn and cfiA was demonstrated (p < 0.000), a positive correlation was found between bfp2 and fpn genes (p = 0.0000803), and a negative correlation between bfp2 and cfiA genes (p = 0.011)