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Mulet M.,University of the Balearic Islands | Gomila M.,Microbiologia | Scotta C.,University of the Balearic Islands | Sanchez D.,University of the Balearic Islands | And 4 more authors.
Systematic and Applied Microbiology | Year: 2012

Multilocus sequence analysis (MLSA) is one of the most accepted methods for the phylogenetic assignation of Pseudomonas strains to their corresponding species. Furthermore, updated databases are essential for correct bacterial identification and the number of Pseudomonas species is increasing continuously. Currently, 127 species are validly described in Euzéby's List of Species with Standing in Nomenclature, and 29 novel species have been described since the publication of the last comprehensive MLSA phylogenetic study based on the sequences of the 16S rDNA, gyrB, rpoB and rpoD genes. Therefore, an update of the sequence database is presented, together with the analysis of the phylogeny of the genus Pseudomonas. Whole-cell matrix-assisted laser-desorption/ionization time-of-flight (WC-MALDI-TOF) mass spectrometry (MS) analysis has been applied very recently to the identification of bacteria and is considered to be a fast and reliable method. A total of 133 type strains of the recognized species and subspecies in the genus Pseudomonas, together with other representative strains, were analyzed using this new technique, and the congruence between the WC-MALDI-TOF MS and MLSA techniques was assessed for the discrimination and correct species identification of the strains. The utility of both methods in the identification of environmental and clinical strains is discussed. © 2012 Elsevier GmbH.

Mineo G.,Radiologia Cardio Toracica | Ciccarese F.,Radiologia I | Modolon C.,Radiologia Pediatrica | Landini M.P.,Microbiologia | And 2 more authors.
Radiologia Medica | Year: 2012

Purpose. Our aim was to evaluate the evolution of 20 patients with H1N1 pneumonia, focusing our attention on patients with severe clinical and radiological findings who developed post-acute respiratory distress syndrome (postARDS) pulmonary fibrosis. Materials and methods. Twenty adult patients (nine women and 11 men; mean age 43.5±16.4 years) with a diagnosis of H1N1 infection confirmed by pharyngeal swab came to our attention from September to November 2009 and were followed up until September 2010. All patients were hospitalised in consideration of the severity of clinical findings, and all underwent chest X-ray. Twelve of them underwent at least one computed tomography (CT) scan of the chest. Results. In 75% of cases (15/20), there was complete resolution of the clinical and radiological findings. Twenty-five percent of patients (5/20) developed acute respiratory distress syndrome (ARDS), which progressed to predominantly peripheral pulmonary fibrosis in 10% (2/20; one died and one had late-onset pulmonary fibrosis, documented on day 68). Moreover, in one patient with a CT diagnosis of pulmonary fibrosis, we observed progressive regression of radiological findings over 4 months of follow-up. Conclusions. In patients with H1N1 pneumonia, postARDS pulmonary fibrosis is not a rare complication. Therefore, a CT scan should be performed in all patients with severe clinical findings. Our study demonstrated that in these patients, fibrosis could present a different spatial distribution and a different temporal trend, with delayed late onset; moreover, in one case, the signs of interstitial lung disease partially regressed over time. Therefore, CT should be considered not only in the diagnostic stage, but also during the follow-up. © 2011 Springer-Verlag.

Bacterial resistance is a frequent and major issue in nosocomial environment. In accordance with the standards proposed by the Clinical and Laboratory Standards Institute (CLSI, 2009), eight strains of Klebsiella pneumoniae (KPC) were isolated from eight distinct patients at Júlia Kubitschek Hospital (JKH), located in the city of Belo Horizonte, Brazil, between April 2009 and January 2011. As far as KPC emergence is concerned, its monitoring in Enterobacteriaceae isolates with decreased susceptibility to ertapenem has become highly important.

Scotta C.,University of the Balearic Islands | Gomila M.,Microbiologia | Mulet M.,University of the Balearic Islands | Lalucat J.,University of the Balearic Islands | And 3 more authors.
Microbial Ecology | Year: 2013

Pseudomonas stutzeri is a widely distributed species with very high genetic diversity and metabolic capacities, occupying many diverse ecological niches. A collection of 229 P. stutzeri strains isolated from different habitats and geographical locations has been previously characterised phylogenetically by rpoD gene sequencing analysis and in the present study 172 of them phenotypically by whole-cell MALDI-TOF mass spectrometry. Fifty-five strains were further analysed by multilocus sequencing analysis to determine the phylogenetic population structure. Both methods showed coherence in strain grouping; 226 strains were allocated in the 18 genomovars known presently. The remaining three strains are proposed as references for three novel genomovars in the species. The correlation and usefulness of sequence-based phylogenetic analysis and whole-cell MALDI-TOF mass spectrometry, which are essential for autoecological studies in microbial ecology, is discussed for the differentiation of P. stutzeri populations. © 2013 Springer Science+Business Media New York.

Cordero-Ampuero J.,Autonomous University of Madrid | Esteban J.,Microbiologia | Garcia-Rey E.,Hospital Universitario La Paz
Clinical Orthopaedics and Related Research | Year: 2010

Background Previous studies of knee arthroplasty infections caused by high-virulence organisms suggest poor outcomes. Polymicrobial and Gram-negative infections are less studied. Questions/purposes This study compared the results of treatment of knee arthroplasty infections by single versus polymicrobial isolates, Gram-positive versus Gram-negative, and methicillin-resistant versus -sensitive Staphylococci. Methods We prospectively followed 47 patients with late knee arthroplasty infections. The mean age was 72 years (range, 20-87 years). The treatment protocol included twostage exchange and a combination of two oral antibiotics given for 6 months. Minimum followup was 1 year (average, 4.8 ± 3 years; range, 1-12 years). Control of the infection was judged by absence of clinical, serologic, and radiologic signs of infection. The functional outcome was evaluated by Knee Society score at the last followup. Results Infection was controlled in all 15 patients with polymicrobial and in 28 of 32 (88%) with monomicrobial infections, in eight of nine patients with Gram-negative and in 35 of 38 (92%) with Gram-positive isolates. Control was also achieved in 22 of 25 patients (88%) infected by methicillin-resistant Staphylococci and in 14 of 14 by methicillin-sensitive Staphylococci. The Knee Society scores averaged 81-63 in patients with polymicrobial infections and were higher than in monomicrobial infections (75-52). The mean KSS was 85-59 in Gram-negative infections compared to 75-55 in Gram-positive infections. The mean KSS was similar in methicillin-resistant (78-54) and methicillin-sensitive Staphylococci (73-56) infections. Conclusions Polymicrobial and Gram-negative infections can be controlled in late knee arthroplasty infections. On the other hand, infections by methicillin-resistant Staphylococci are less likely to be controlled by the regimens we used. Level of Evidence Level II, prognostic study. See Guidelines for Authors for a complete description of levels of evidence. © The Association of Bone and Joint Surgeons® 2010.

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