Paracoccidioidomycosis molecular diagnosis associated with tuberculosis in sputum samples

Introdução: A paracoccidioidomicose (PCM) é uma micose sistêmica endêmica causada pelo fungo Paracoccidioides spp. O objetivo deste estudo foi determinar a associação entre tuberculose (TB) e PCM em pacientes com exame micológico negativo. Métodos: Estudo prospectivo de diagnóstico molecular de amostras de escarro, com resultado positivo para bacilo álcool ácido resistente (BAAR) e negativo no exame direto e cultivo micológico. Resultados: A aplicação de técnicas moleculares resultou em 18,4% de pacientes coinfectados com PCM e TB. Conclusão: O conhecimento das diferenças clínicas, epidemiológicas e laboratoriais da PCM quando associada à TB é importante para prevenir a disseminação da doença, complicações e o aumento da letalidade.Introduction: Paracoccidioidomycosis (PCM) is a systemic endemic mycosis caused by Paracoccidioides spp. The aim of this study was to determine the association between tuberculosis (TB) and PCM in patients with negative mycological examination results. Methods: Prospective study of molecular diagnosis of sputum samples, with positive results for bacilli resistant acid (BAAR) and negative results on direct examination and mycological culture. Results: The application of molecular techniques resulted in 18.4% of patients co-infected with PCM and TB. Conclusion: The knowledge of clinical, epidemiological, and laboratory differences of PCM when associated with TB is important to prevent the spread of disease, complications, and increased mortality

Molecular epidemiology of heteroresistant vancomycin-intermediate Staphylococcus aureus in Brazil

AbstractTo determine the epidemiological and molecular characteristics of 12 Staphylococcus aureus isolates presenting heteroresistance to vancomycin in laboratories of two cities in Santa Catarina, southern Brazil. Epidemiological data, including the city of isolation, health institution, and date of isolation were considered, as well as the associated clinical specimen. For molecular characterization, we analyzed the staphylococcal cassette chromosome types, the erm gene presence, and the genomic diversity of isolates using pulsed-field gel electrophoresis. The 12 isolates of S. aureus were previously confirmed as heteroresistance to vancomycin using the population analysis profile–area under curve. Regarding genetic variability, two clones were detected: the main one (clone A) composed of four isolates and the clones B, with two isolates. For clone A, two isolates presented identical band patterns and were related to the same hospital, with an interval of 57 days between their isolation. The other isolates of this clone showed no epidemiological link between them because they were isolated in different hospitals and had no temporal relationship. The other clone showed no detectable epidemiological relationship. The heteroresistance to vancomycin recovered in Santa Catarina State from 2009 to 2012 had, in general, heterogeneous genomic patterns based on pulsed-field gel electrophoresis results, which is in accordance with the fact that these isolates had little or no epidemiological relationship among them. Due to the characteristic phenotypic instability and often prolonged vancomycin therapy for selection, clonal spread is not as common as for other resistance mechanisms disseminated through horizontal gene transfer

Susceptibility profiles and correlation with pneumococcal serotypes soon after implementation of the 10-valent pneumococcal conjugate vaccine in Brazil

SummaryObjectivesTo evaluate the susceptibility patterns among Streptococcus pneumoniae recovered during the years 2010–2012 and to correlate these with serotypes.MethodsPneumococci from invasive sites were serotyped by sequential multiplex PCR and/or Quellung reaction. Etest strips were used to determine the minimal inhibitory concentrations, and the Clinical and Laboratory Standards Institute (CLSI) guidelines were used for interpretation. Genetic determinants of macrolide resistance were assessed by PCR, and the occurrence of the D phenotype was analyzed following the recommendations of the CLSI.ResultsOne hundred fifty-nine S. pneumoniae were studied; most were recovered from blood and were associated with serotypes 14, 3, 4, 23F, 20, 7F, 12F, 19A, and 19F. Pneumococcal conjugate vaccine PCV7, PCV10, and PCV13 and 23-valent polysaccharide vaccine serotypes represented 38.2%, 48.7%, 64.5%, and 85.5%, respectively. β-Lactam non-susceptibility (non-meningitis) was basically related to serotype 19A. For meningitis, it was observed in 21.4% (serotypes 14, 3, 9V, 23F, and 24F). Resistance to erythromycin occurred in 8.2% and mefA was the most common macrolide genetic determinant. One isolate was resistant to levofloxacin. Non-susceptibility to trimethoprim–sulfamethoxazole was 37.7% and to tetracycline was 22.0%.ConclusionsOur population of pneumococci represents a transition era, soon after the introduction of PCV10. Non-susceptible patterns were found to be associated with classical PCV serotypes (especially serotype 14), which is still highly prevalent, and non-PCV10 ones (19A), which may disseminate, occupying the biological niche left by the vaccine serotypes

Coexistence of virulence genes in methicillin-resistant Staphylococcus aureus clinical isolates

Abstract INTRODUCTION: The pathogenic versatility of Staphylococcus aureus is attributed to various virulence genes, including enterotoxins and hemolysins. METHODS: Here, the virulence genes in 177 nosocomial MRSA strains in Porto Alegre, Brazil were detected by PCR. RESULTS: The overall prevalence rates were as follows: sea, 4.5%; pvl, 18.6%; tst, 27.7%; hla, 87.6%; and hld, 90.4%. No strain contained all tested genes. However, there was frequent coexistence of tst with pvl and hla with hld (40.7% and 26.6%, respectively). CONCLUSIONS: Horizontal transfer of virulence genes is very common in S. aureus, as suggested by the frequent coexistence of several virulence genes

Paracoccidioidomycosis molecular diagnosis associated with tuberculosis in sputum samples

Introdução: A paracoccidioidomicose (PCM) é uma micose sistêmica endêmica causada pelo fungo Paracoccidioides spp. O objetivo deste estudo foi determinar a associação entre tuberculose (TB) e PCM em pacientes com exame micológico negativo. Métodos: Estudo prospectivo de diagnóstico molecular de amostras de escarro, com resultado positivo para bacilo álcool ácido resistente (BAAR) e negativo no exame direto e cultivo micológico. Resultados: A aplicação de técnicas moleculares resultou em 18,4% de pacientes coinfectados com PCM e TB. Conclusão: O conhecimento das diferenças clínicas, epidemiológicas e laboratoriais da PCM quando associada à TB é importante para prevenir a disseminação da doença, complicações e o aumento da letalidade.Introduction: Paracoccidioidomycosis (PCM) is a systemic endemic mycosis caused by Paracoccidioides spp. The aim of this study was to determine the association between tuberculosis (TB) and PCM in patients with negative mycological examination results. Methods: Prospective study of molecular diagnosis of sputum samples, with positive results for bacilli resistant acid (BAAR) and negative results on direct examination and mycological culture. Results: The application of molecular techniques resulted in 18.4% of patients co-infected with PCM and TB. Conclusion: The knowledge of clinical, epidemiological, and laboratory differences of PCM when associated with TB is important to prevent the spread of disease, complications, and increased mortality

Enterococcus species diversity in fecal samples of wild marine species by real-time PCR

Analyses using culture-independent molecular techniques have improved our understanding of microbial composition. The aim of this work was to identify and quantify enterococci in fecal samples of wild marine species using real-time quantitative PCR (qPCR). Seven Enterococcus species were examined in fecal DNA of South American fur seals (Arctocephalus australis), Subantarctic fur seals (Arctocephalus tropicalis), green turtles (Chelonia mydas), Magellanic penguins (Spheniscus magellanicus), snowy-crowned Tern (Sterna trudeaui), white-backed Stilt (Himantopus melanurus), white-chinned Petrels (Procellaria aequinoctialis), red knot (Calidris canutus), black-browed albatross (Thalassarche melanophris). All Enterococcus species evaluated were detected in all fecal samples of wild marine species, with a concentration ranged between 106 to 1012 copies/ng of total DNA. Differences in the enterococci distribution were observed. Enterococcus faecalis and E. mundtii were most abundant in marine mammals. Enterococcus faecalis was frequent in green turtles, Magellanic penguins, snowy-crowned Tern, red knot, black-browed and albatross. Enterococcus hirae and E. gallinarum showed elevated occurrence in white-backed Stilt, and E. faecium in white-chinned Petrel. This study showed highest diversity of enterococci in feces of wild marine species than currently available data, and ensure that culture-independent analysis help us to enhance our understanding about enterococci in gastrointestinal tracts of wild marine species.The accepted manuscript in pdf format is listed with the files at the bottom of this page. The presentation of the authors' names and (or) special characters in the title of the manuscript may differ slightly between what is listed on this page and what is listed in the pdf file of the accepted manuscript; that in the pdf file of the accepted manuscript is what was submitted by the author

Serotypes and Genotypes of Invasive <i>Streptococcus pneumoniae</i> Before and After PCV10 Implementation in Southern Brazil

<div><p>To reduce the burden of pneumococcal diseases, different formulations of pneumococcal conjugate vaccines (PCV) have been introduced in many countries. In Brazil, PCV10 has been available since 2010. We aimed to analyze the serotype and genetic composition of invasive pneumococci from Brazil in pre- and post- vaccination periods (2007–2012). Antibiotic susceptibility was determined and genotypes of macrolide and fluoroquinolone resistance were characterized. The genotypes of isolates of the most frequent serotypes were determined by multilocus sequence typing. The study included 325 isolates, which were primarily recovered from blood. The most common serotypes recovered were 14, 3, 4, 23F, 7F, 9V, 12F, 20, 19F, 8, 19A, and 5. Thirty-eight pneumococci (11.7%) were from children ≤5 years old. Considering the overall population, PCV10 and PCV13 serotype coverage was 50.1% and 64.9%, respectively. During the pre-vaccine period, isolates with serotypes belonging to the PVC10 represented 51.5% (100/194), whereas in the post vaccine they represented 48.0% (63/131). PCV13 serotypes represented 67.5% (131/194) and 59.2% (77/131) of total for pre- and post-vaccination periods, respectively. Seventy different sequence types [STs] were found, accounting for 9 clonal complexes [CCs] and 45 singletons. Eight STs (156, 180, 218, 8889, 53, 191, 770, and 4967) represented the majority (51.5%) of isolates. Fifty STs were associated with the pre-vaccination period (27 exclusive) and 43 (20 exclusive) with the post-vaccination period; 23 STs were identified in both periods. Some serotypes were particularly clonal (7F, 8, 12F, 20). Non-susceptibility to penicillin was associated with serotype 19A, CC320. Erythromycin resistance was heterogeneous when considering serotype and ST. A single serotype 23F (ST4967) isolate was resistant to levofloxacin. Continued surveillance is required to determine vaccine impact and to monitor changes in pneumococcal population biology post-PCV10 introduction in Brazil.</p></div

Serotypes and STs observed among 231 pneumococci recovered from IPD (2007–2012).

<p>Serotypes and STs observed among 231 pneumococci recovered from IPD (2007–2012).</p

Distribution of serotypes belonging to PCV10 among invasive pneumococci recovered from 2007 to 2012 (the distribution of all serotypes is demonstrated on the Table S1).

<p>*Pre-vaccination period;</p><p>**post-vaccination period;</p>#<p>OR: odds ratio.</p><p>Distribution of serotypes belonging to PCV10 among invasive pneumococci recovered from 2007 to 2012 (the distribution of all serotypes is demonstrated on the <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0111129#pone.0111129.s001" target="_blank">Table S1</a>).</p