Diverse sequence types of Klebsiella pneumoniae contribute to the dissemination of bla NDM-1 in India, Sweden, and the United Kingdom

Clinical isolates of Klebsiella pneumoniae producing NDM-1 carbapenemase from India (n = 22), the United Kingdom (n = 13), and Sweden (n = 4) were subjected to multilocus sequence typing (MLST), automated repetitive sequence-based PCR (rep-PCR), serotyping, virulence gene screening, and plasmid replicon typing. The most frequently detected MLST sequence types (STs) were ST14 (n = 13; all serotype K2), ST11, ST149, ST231, and ST147. The correlation between MLST and automated rep-PCR was excellent. IncA/C was the most frequently detected plasmid replicon type (n = 14). ST14, ST11, and other successful clones may be important for the dissemination of bla . Copyrigh

Prevalence and morphological variability of torus palatanus and torus mandibularis

Background: The oral tori are non-pathological and benign exostosis of the cortical and limited amount of bone marrow, covered with a thin and poorly vascularized mucosa. The purpose of this study was to determine the prevalence of torus palatanus (TP) and torus mandibularis (TM) and to define morphological variability in relation to age and gender in the population of Shtip region, Macedonia FYR. Methods and materials: The study comprised of 467 patients, 242 females and 225 males, from 20 years old and onwards who were examined by clinical examination and analysis of the plaster casts. Results: Torus palatinus was found in 10.9% subjects and torus mandibularis in 9.6% of the subjects. The torus palatinus was well developed in 48.4 % of the individuals, nodular form and its more frequent location was in the middle palate. The most frequent type of torus mandibularis was bilateral solitary torus mandibularis (32.4%). The results of this study show significantly higher prevalence of torus palatinus and torus mandibularis in the male subjects. Conclusion: The results of this study show significantly relationship between the occurrence of tori and gender and although not significant, there is a trend towards higher prevalence of TP and TM with increasing age

High Genetic Diversity among Community-Associated Staphylococcus aureus in Europe: Results from a Multicenter Study

Background: Several studies have addressed the epidemiology of community-associated Staphylococcus aureus (CA-SA) in Europe; nonetheless, a comprehensive perspective remains unclear. In this study, we aimed to describe the population structure of CA-SA and to shed light on the origin of methicillin-resistant S. aureus (MRSA) in this continent. Methods and Findings: A total of 568 colonization and infection isolates, comprising both MRSA and methicillin-susceptible S. aureus (MSSA), were recovered in 16 European countries, from community and community-onset infections. The genetic background of isolates was characterized by molecular typing techniques (spa typing, pulsed-field gel electrophoresis and multilocus sequence typing) and the presence of PVL and ACME was tested by PCR. MRSA were further characterized by SCCmec typing. We found that 59 % of all isolates were associated with community-associated clones. Most MRSA were related with USA300 (ST8-IVa and variants) (40%), followed by the European clone (ST80-IVc and derivatives) (28%) and the Taiwan clone (ST59-IVa and related clonal types) (15%). A total of 83 % of MRSA carried Panton-Valentine leukocidin (PVL) and 14 % carried the arginine catabolic mobile element (ACME). Surprisingly, we found a high genetic diversity among MRSA clonal types (ST-SCCmec), Simpson’s index of diversity = 0.852 (0.788–0.916). Specifically, about half of the isolates carried novel associations between genetic background and SCCmec. Analysis by BURP showed that some CA-MSSA and CA-MRS

PLoS ONE

Background and Aims Neonatal infections caused by Extended-spectrum beta-lactamase (ESBL)-producing bacteria are associated with increased morbidity and mortality. No data are available on neonatal colonization with ESBL-producing bacteria in Ecuador. The aim of this study was to determine the proportion of intestinal colonization with ESBL-producing Enterobacteriaceae, their resistance pattern and risk factors of colonization in a neonatal intensive care unit in Ecuador. Methods During a three month period, stool specimens were collected every two weeks from hospitalized neonates. Species identification and susceptibility testing were performed with Vitek2, epidemiologic typing with automated repetitive PCR. Associations between groups were analyzed using the Pearson X2 test and Fisher exact test. A forward step logistic regression model identified significant predictors for colonization. Results Fifty-six percent of the neonates were colonized with ESBL-producing Enterobacteriaceae. Length of stay longer than 20 days and enteral feeding with a combination of breastfeeding and formula feeding were significantly associated with ESBL-colonization. The strains found were E. coli (EC, 89%) and K. pneumoniae (KP, 11%) and epidemiological typing divided these isolates in two major clusters. All EC and KP had blaCTX-M group 1 except for a unique EC isolate that had blaCTX-M group 9. Multi-locus sequence typing performed on the K. pneumoniae strains showed that the strains belonged to ST855 and ST897. The two detected STs belong to two different epidemic clonal complexes (CC), CC11 and CC14, which previously have been associated with dissemination of carbapenemases. None of the E. coli strains belonged to the epidemic ST 131 clone. Conclusions More than half of the neonates were colonized with ESBL-producing Enterobacteriaceae where the main risk factor for colonization was length of hospital stay. Two of the isolated clones were epidemic and known to disseminate carbapenemases. The results underline the necessity for improved surveillance and infection control in this context.Cuencavolumen 8; número 1

Microfluidic system for the identification of bacterial pathogens causing urinary tract infections

Urinary tract infections (UTIs) are among the most common bacterial infections and pose a significant healthcare burden. The growing trend in antibiotic resistance makes it mandatory to develop diagnostic kits which allow not only the determination of a pathogen but also the antibiotic resistances. We have developed a microfluidic cartridge which takes a direct urine sample, extracts the DNA, performs an amplification using batch-PCR and flows the sample over a microarray which is printed into a microchannel for fluorescence detection. The cartridge is injection-molded out of COP and contains a set of two-component injection-molded rotary valves to switch between input and to isolate the PCR chamber during thermocycling. The hybridization probes were spotted directly onto a functionalized section of the outlet microchannel. We have been able to successfully perform PCR of E. coli in urine in this chip and perform a fluorescence detection of PCR products. An upgraded design of the cartridge contains the buffers and reagents in blisters stored on the chip.QC 20150617</p

Four genotyping schemes for phylogenetic analysis of Pseudomonas aeruginosa: comparison of their congruence with multi-locus sequence typing.

Several molecular typing schemes have been proposed to differentiate among isolates and clonal groups, and hence establish epidemiological or phylogenetic links. It has been widely accepted that multi-locus sequence typing (MLST) is the gold standard for phylogenetic typing/long-term epidemiological surveillance, but other recently described methods may be easier to carry out, especially in settings with limited access to DNA sequencing. Comparing the performance of such techniques to MLST is therefore of relevance. A study was therefore carried out with a collection of P. aeruginosa strains (n = 133) typed by four typing schemes: MLST, multiple-locus variable number tandem repeat analysis (MLVA), pulsed-field gel electrophoresis (PFGE) and the commercial DiversiLab microbial typing system (DL). The aim of this study was to compare the results of each typing method with MLST. The Simpson's indices of diversity were 0.989, 0.980, 0.961 and 0.906 respectively for PFGE, MLVA, DL and MLST. The congruence between techniques was measured by the adjusted Wallace index (W): this coefficient indicates the probability that a pair of isolates which is assigned to the same type by one typing method is also typed as identical by the other. In this context, the congruence between techniques was recorded as follow: MLVA-type to predict MLST-type (93%), PFGE to MLST (92%), DL to MLST (64.2%), PFGE to MLVA (63.5%) and PFGE to DL (61.7%). Conversely, for all above combinations, prediction was very poor. The congruence was increased at the clonal complex (CC) level. MLST is regarded the gold standard for phylogenetic classification of bacteria, but is rather laborious to carry out in many settings. Our data suggest that MLVA can predict the MLST-type with high accuracy, and even higher when studying the clonal complex level. Of the studied three techniques MLVA was therefore the best surrogate method to predict MLST

Minimal Spanning Tree of <i>K. pneumoniae</i> Isolates in Fecal Carriage in Children in Guinea-Bissau.

<p>Full legend: The tree maps the relatedness of <i>K. pneumoniae</i> isolates. Isolates ≥93% related in DiversiLab analysis are grouped in a pie where each slice represents one isolate in the cluster. Each cluster was assigned a Roman numeral. Red indicates resistance to gentamicin, ciprofloxacin and trimethoprim-sulfamethoxazole, which at the time of the study were all easily available antibiotics except cephalosporin to treat gram-negative bacterial infections in Guinea-Bissau. Green indicates susceptibility to at least one of the mentioned agents.</p

Minimal Spanning Tree of <i>E. coli</i> Isolates in Fecal Carriage in Children in Guinea-Bissau.

<p>Full legend: The tree maps the relatedness of <i>E. coli</i> isolates. Isolates ≥95% related in DiversiLab analysis are grouped in a pie where each slice represents one isolate in the cluster. Each cluster was assigned a Roman numeral. Red indicates resistance to gentamicin, ciprofloxacin and trimethoprim-sulfamethoxazole, which at the time of the study were all easily available antibiotics except cephalosporin to treat gram-negative bacterial infections in Guinea-Bissau. Green indicates susceptibility to at least one of the mentioned agents.</p

Study Population Characteristics and Risk Factors for Colonization with ESBL-Producing Bacteria.

1<p>Mid-upper arm circumference at time of enrolment examined on children ≥6 months of age.</p>2<p>Test for linear trend.</p>3<p>Child breastfed at time of enrolment.</p>4<p>Bedsharing with another child <5 years of age.</p>5<p>Number of children <5 years of age living in the same household.</p>6<p>Antibiotic treatment initiated prior to presentation at emergency ward.</p>7<p>Reported antibiotic usage during the month prior to study enrolment (excluding antibiotic usage for current disease).</p>8<p>Child hospitalized ≥1 day during the month prior to enrolment.</p