Carbapenemase-Producing Klebsiella pneumoniae in Romania: A Six-Month Survey.

This study presents the first characterization of carbapenem-non-susceptible Klebsiella pneumoniae isolates by means of a structured six-month survey performed in Romania as part of an Europe-wide investigation. Klebsiella pneumoniae clinical isolates from different anatomical sites were tested for antibiotic susceptibility by phenotypic methods and confirmed by PCR for the presence of four carbapenemase genes. Genome macrorestriction fingerprinting with XbaI was used to analyze the relatedness of carbapenemase-producing Klebsiella pneumoniae isolates collected from eight hospitals.

Molecular characterization of bacteremic Escherichia coli isolates in Romania.

The increasing prevalence of invasive infections caused by antibiotic resistant Escherichia coli strains in Romanian patients, already mentioned in the European reports, requires better knowledge of their specific traits. Thus, a set of 38 E. coli blood isolates, collected between 2010 and 2012 at one of the local hospitals participating into the European Antimicrobial Resistance Surveillance Network, was investigated retrospectively with respect to the phylogenetic origin, extraintestinal virulence-associated markers (i.

MOLECULAR CHARACTERIZATION OF STAPHYLOCOCCUS AUREUS STRAINS ISOLATED FROM INFECTIVE ENDOCARDITIS.

Infective endocarditis (IE) is an infection of the heart endothelium and valves and is frequently a consequence of a sanguine flow turbulence and injury of endocardium. Recent studies revealed an increase of Staphylococcus aureus strains involved in IE, but no evident correlations between the genetic background of this bacterium and IE involvement of certain strains have been found yet. In this study we analyzed the virulence profile, including adhesins, exotoxins, superantigens and biofilm determinants, along with agr type detection, for S.

Escherichia coli ST131 causing invasive infections in Romanian patients--a threat we can no longer ignore.

Escherichia coli sequence type ST131 is a major pandemic clonal group of drug-resistant extraintestinal pathogenic E. coli (ExPEC) involved in community-onset and healthcare-associated infections. Thus far, its presence in our area has been paid little attention.

Genetic diversity and antimicrobial resistance in Streptococcus agalactiae strains recovered from female carriers in the Bucharest area.

For the first time, we used multilocus sequence typing (MLST) to understand how Romanian group B streptococcus (GBS) strains fit into the global GBS population structure. Colonising isolates recovered from adult human females were tested for antibiotic resistance, were molecularly serotyped based on the capsular polysaccharide synthesis (cps) gene cluster and further characterised using a set of molecular markers (surface protein genes, pilus-encoded islands and mobile genetic elements inserted in the scpB-lmb intergenic region). Pulsed-field gel electrophoresis was used to complement the MLST clonal distribution pattern of selected strains.

Rapid identification of international multidrug-resistant Pseudomonas aeruginosa clones by multiple-locus variable number of tandem repeats analysis and investigation of their susceptibility to lytic bacteriophages.

The objective of this study was to determine the genetic diversity of multidrug-resistant (MDR) Pseudomonas aeruginosa strains isolated over a period of 12 months in two French hospitals and to test their susceptibility to bacteriophages. A total of 47 MDR isolates recovered from hospitalized patients were genotyped using multiple-locus variable number of tandem repeats analysis. The genotypes were distributed into five clones (including 19, 5, 5, 3, and 3 isolates, respectively) and 12 singletons.

Recurrent Infective endocarditis of the native aortic valve due to ESBL producing Escherichia coli (e coli) after therapeutic ERCP.

Unlabelled: We describe the first case of ESBL producing E coli endocarditis of the native aortic valve in which prophylaxis was performed according to currently available guidelines. A 75 year-old woman presented at our emergency department with a two week complaint of fever, fatigue, anorexia, diffuse abdominal pain after ERCP therapeutic. The initial laboratory examinations showed increased levels of ESR, CRP, fibrinogen, alkaline phosphatase, gammaGT, aspartate aminotransferase, alanine aminotransferase and direct fraction of bilirubin.

Methodology optimization and diversification for the investigation of virulence potential in Haemophilus influenzae clinical strains.

Ten Haemophilus influenzae strains were isolated from patients aged between 1.6 - 24 years, with various diagnoses (acute meningitis, acute upper respiratory infection, otitis media and acute sinusitis). Identification was based on phenotypic and molecular characteristics; antibiotic susceptibility testing was performed by diffusion method according to CLSI standards 2011 for seven antibiotics.

Molecular characterization of adult-colonizing Streptococcus agalactiae from an area-based surveillance study in Romania.

One hundred and forty-eight colonizing isolates from adult Romanian women were conventionally serotyped and screened for antibiotic resistance. Capsular type assignment by multiplex polymerase chain reaction (PCR) was performed for nonserotypeable isolates. Tetracycline and macrolide resistance genes (tetM, tetO, tetL, ermA, ermB, and mefA) including tetM gene association with conjugative elements of the Tn916 family were searched.

Q fever endocarditis in Romania: the first cases confirmed by direct sequencing.

Infective endocarditis (IE) is a serious, life-threatening disease with highly variable clinical signs, making its diagnostic a real challenge. A diagnosis is readily made if blood cultures are positive, but in 2.5 to 31% of all infective endocarditis cases, routine blood cultures are negative.

A multiple typing approach is recommended for the laboratory-based surveillance of salmonellosis in Romania.

In Romania, Salmonella enterica serovar Typhimurium isolates are currently typed by antimicrobial resistance profiles and phage typing, as part of the national laboratory-based surveillance system of human enteric infections. The aim of the present study was to assess the added value of complementing this approach with molecular fingerprinting, namely pulsed-field gel electrophoresis (PFGE) and multiple-locus variable-number tandem-repeats analysis (MLVA). Thirty-six S.

Group B streptococcus colonization of Romanian women: phenotypic traits of isolates from vaginal swabs.

In the attempt to enrich the local contemporary laboratory data regarding the group B streptococcus (GBS) colonization, isolates obtained from the vaginal swab cultures were characterized for their serotype distribution and antibiotic susceptibility. The 100 GBS isolates analyzed were collected during a four-month period of year 2009 from women screened in ambulatory for vaginal carriage of GBS. The GBS isolates were classified based on their capsular polysaccharide structures using commercially available antisera.

Clinical and microbiological characteristics of severe Streptococcus pyogenes disease in Europe.

In an attempt to compare the epidemiology of severe Streptococcus pyogenes infection within Europe, prospective data were collected through the Strep-EURO program. Surveillance for severe cases of S. pyogenes infection diagnosed during 2003 and 2004 was undertaken in 11 countries across Europe by using a standardized case definition and questionnaire.

Molecular characterization of invasive and non-invasive Streptococcus pyogenes isolates from Romania.

In 2002, the Romanian National Reference Laboratory was invited to join the Strep-EURO project to study invasive Streptococcus pyogenes infections. During 2003 and 2004, a total of 33 isolates recovered from invasive disease were received from eight Romanian counties. For comparison, 102 isolates from non-invasive disease, as well as a collection of 12 old invasive strains (isolated between 1967 and 1980) were included.

Regulatory mechanisms differ in UMP kinases from gram-negative and gram-positive bacteria.

In this work, we examined the regulation by GTP and UTP of the UMP kinases from eight bacterial species. The enzyme from Gram-positive organisms exhibited cooperative kinetics with ATP as substrate. GTP decreased this cooperativity and increased the affinity for ATP.

Comparative study of three different DNA fingerprint techniques for molecular typing of Shigella flexneri strains isolated in Romania.

In this study, 97 epidemiologically unrelated Shigella flexneri strains isolated during 1994 (69 isolates) and 1997 (28 isolates) were characterised by ribotyping, enterobacterial repetitive intergenic consensus sequence-based PCR typing, and pulsed-field gel electrophoresis. Number of strains belonging to each of the six serotypes is selected equal to their distribution in Romania. The isolates comprise 24 ribotypes based on combination of two restriction patterns obtained with HindlII and PstI, respectively, 7 enterobacterial repetitive intergenic consensus (ERIC)-PCR types, and 92 XbaI pulsed-field gel electrophoresis (PFGE) patterns grouped in 31 pulsotypes at Dice coefficients of 85% similarity.

Study of Corynebacterium diphtheriae strains isolated in Romania, northwestern Russia and the Republic of Moldova.

A selection of 167 Corynebacterium diphtheriae strains isolated in Romania, the Russian Federation and the Republic of Moldova were analysed by biotyping, phage typing, the toxin production test and by molecular techniques such as ribotyping, pulsed field gel electrophoresis and random amplified polymorphic DNA, in order to establish the epidemiological relatedness, genetic divergence and strain circulation within and between the bordering countries. Using a set of five digoxigenin-labeled oligonucleotides and BstEII digestion, 34 ribotypes were identified. The strains isolated in the epidemic areas (Russia and Moldova) were very closely related but different from those isolated in Romania.

Correlation between two DNA-based typing methods applied to Streptococcus pneumoniae strains: BOX-PCR assay as an alternative to ribotyping.

Seventy-nine Streptococcus pneumoniae isolates were characterized by ribotyping and BOX-PCR typing. The isolates comprised 14 most frequently identified serogroups and serotypes, 54 ribotypes (defined as each different combination of two DNA restriction fragment length polymorphism patterns obtained with HindIII and PvuII, respectively) and 51 BoxA-PCR patterns. There was close but not absolute correlation between molecular techniques.

Molecular subtyping of group A streptococcal strains based on virulence regulon polymorphism.

A total of 64 Streptococcus pyogenes isolates included in a previous study (Shundi et al., 2000) were further analyzed by Vir typing. Vir typing is based on restriction fragment length polymorphism (RFLP) analysis of a 4- to 8-kb pathogenicity island (vir regulon) encoding emm and other virulence genes.

Molecular typing of Enterococcus faecalis strains resistant to high levels of gentamicin and isolated in Romania.

Sixteen Enterococcus faecalis strains resistant to high levels of gentamicin, 15 of which were isolated in the same year in a Romanian hospital, harboured conjugative gentamicin resistance (Gm(r)) plasmids ranging from 55 to 85 kilobases. On the basis of restriction enzyme and DNA-DNA hybridization profiles of these plasmids, as well as of chromosomal SmaI macrorestriction and Tn916 hybridization patterns, clonal relationship was established for seven strains whereas the other strains were considered to be independent. Nine and seven of the Gm(r) plasmids carried Tn4001-like and Tn4001-truncated structures, respectively; the latter structures were truncated in the right-hand flanking extremity of the element.

Plasmid-borne high-level resistance to gentamicin in Enterococcus hirae, Enterococcus avium, and Enterococcus raffinosus.

Enterococcus hirae, E. avium, and E. raffinosus isolated in Romania, Tunisia, and Portugal harbored plasmids pICC8, pIP1700, and pIP1701, respectively, encoding resistance to high levels of gentamicin (Gmr).