Detection of cross-infection associated to a Brazilian PCR-ribotype of Clostridium difficile in a university hospital in Rio de Janeiro, Brazil.

Clostridium difficile is an important nosocomial enteric pathogen and is the etiological agent of pseudomembranous colites. Recently, the rates of C. difficile infection (CDI) have increased worldwide, but in Brazil few data about this situation and the incidence of clonal types of C. difficile exist. This study aimed to isolate and characterize C. difficile strains from samples obtained of a university hospital (HUCFF) in Rio de Janeiro city, Brazil. CDI was identified by ELISA in 27.1% of HUCFF-in-patients enrolled in the study, and the bacterium was recovered from eight of these fecal samples. All strains, except one, presented tcdA and tcdB genes and presented neither the cdtA and cdtB genes nor any significant deletions in the tcdC gene. All strains were sensitive to metronidazole, vancomycin and moxifloxacin, and resistant to clindamycin, ciprofloxacin and levofloxacin. PCR-ribotyping and PFGE revealed four different clonal types among the isolates. The Brazilian PCR-ribotype 133 accounted for 50% of strains isolated, and PCR-ribotype 233 strains were obtained from 25% of the in-patients. The prevalence and resurgence of the Brazilian PCR-ribotype 133 among the hospitalized patients of HUCFF was established, and cross-infection of different patients associated to the same PCR-ribotypes was detected. Our results emphasize the importance of the diagnosis and control of CDI in order to prevent the emergence of specific clones that can lead to C. difficile-associated outbreaks in Brazilian hospitals.

Draft Genome Sequences of Leptospira interrogans Serovar Copenhageni Strains Isolated from Patients with Weil's Disease in Brazil.

Leptospirosis is a worldwide zoonosis caused by pathogenic species of Leptospira In Brazil, this disease is endemic, presenting epidemic potential in rainy seasons. Here, we announce the whole-genome sequences of two L. interrogans serovar Copenhageni strains isolated from blood samples from two icteric patients associated with severe leptospirosis in Brazil.

Characterization of Clostridium difficile strains isolated from immunosuppressed inpatients in a hospital in Rio de Janeiro, Brazil.

The aim of this work was to identify and characterize Clostridium difficile strains from fecal and hospital environmental samples. C. difficile toxins were detected by ELISA in 28.5% of the analyzed samples. Four strains were isolated from immunosuppressed inpatients presenting antibiotic-associated diarrhea. All strains possessed tcdA and tcdB genes and did not present neither the cdtA and cdtB genes nor any significant deletions in the tcdC gene. PFGE and PCR-ribotyping analysis showed that two strains belonged to the same clonal type (ribotype 014) and the other two were grouped into ribotype 106, in spite of presenting a similar, but not identical genetic fingerprint. This report shows that for the first time ribotype 106 was found outside the United Kingdom. All isolates were equally sensitive to metronidazole. The ribotype 014 isolates were highly resistant to clindamycin, while the ribotype 106 isolates were resistant to all fluoroquinolones tested. This work reveals the spread of C. difficile in the hospital unit studied and the presence of three genetically related types, two of them presenting resistance to fluoroquinolones.

Enterotoxigenic and nontoxigenic Bacteroides fragilis strains isolated in Brazil.

The presence of enterotoxigenic Bacteroides fragilis and nontoxigenic B. fragilis (NTBF) among 109 strains isolated from 1980-2008 in Brazil were investigated by PCR. One strain, representing 0.9% of the total analyzed strains, harbored the bft gene which was identified as bft-1 isoform based on PCR-RFLP and sequencing. Forty-nine strains (44.9%) exhibited the NTBF pattern III which possesses the flanking region required for pathogenicity island acquisition in which the bft gene is codified. These data reinforce the potential of B. fragilis as an emerging enteropathogen in our country.

New PCR ribotypes of Clostridium difficile detected in children in Brazil: prevalent types of Clostridium difficile in Brazil.

A total of 35 Brazilian isolates of Clostridium difficile from faecal stools and four isolates from hospital environments were analyzed by PCR ribotyping. A whole cell protein profile (as an alternative for serogrouping), in vitro toxin production and susceptibility to vancomycin, metronidazole and clindamycin were also investigated. All strains were typeable by both phenotypic and genotypic methods, and a total of 13 different PCR ribotypes were identified, of which seven (132, 133, 134, 135, 136, 142 and 143) were considered new types and accounted for 78.5% of all samples evaluated (including hospital environments). A non-toxigenic C. difficile PCR ribotype 133 was detected in all children groups examined (inpatients, outpatients and healthy children), whilst toxigenic PCR ribotypes 015, 131, 134 and 135 were associated mostly with symptomatic children. Serogroups G and D were disseminated both in patients from the community and from the pediatric hospital, with group G prevalent among outpatient children. All strains were susceptible to vancomycin and metronidazole but high levels of resistance to clindamycin were found, especially among serogroups G and D. Co-existence of different ribotypes and serogroups in the same individual was observed. The new seven ribotypes found in this investigation may represent strains characteristic of this region of Brazil.

Enterotoxigenic and nontoxigenic Bacteroides fragilis strains isolated in Brazil

The presence of enterotoxigenic Bacteroides fragilis and nontoxigenic B. fragilis (NTBF) among 109 strains isolated from 1980-2008 in Brazil were investigated by PCR. One strain, representing 0.9 percent of the total analyzed strains, harbored the bft gene which was identified as bft-1 isoform based on PCR-RFLP and sequencing. Forty-nine strains (44.9 percent) exhibited the NTBF pattern III which possesses the flanking region required for pathogenicity island acquisition in which the bftgene is codified. These data reinforce the potential of B. fragilis as an emerging enteropathogen in our country.