Incidence and importance of Clostridium difficile in paediatric diarrhoea in Brazil.

Clostridium difficile strains were detected in 14 of 210 (6.7 %) faecal samples from children in Rio de Janeiro, Brazil, by cultivating faeces on cycloserine/cefoxitin/fructose agar after alcohol-shock. Two main groups of children were studied: inpatients (n = 96) and outpatients (n = 114). The inpatient group consisted of children on antibiotics or immunosuppressors who presented with diarrhoea and other children who did not present with diarrhoea and were not under an antibiotic or chemotherapeutic regimen. Among the outpatients, two groups were examined: namely, a group that comprised children who presented with diarrhoea and were occasionally under an antibiotic regimen and another group that comprised patients who were not taking antibiotics. After cytotoxic assay, toxigenic C. difficile (Cd tox+) strains were detected in 4.2 % of inpatients and 3.5 % of outpatients. Exclusion of other infectious causes of diarrhoea indicated a typical case of C. difficile-associated paediatric diarrhoea in the community. Among Cd tox+ isolates, no variations were detected by PCR for toxin A that employed primers NK9 and NKVO11. No resistance was found to metronidazole or vancomycin among strains that were isolated from children who presented with diarrhoea, but the MIC(50) and MIC(90) values for clindamycin were 6-8 and 16 microg ml(-1), respectively. Resistance to clindamycin seems to be more disseminated in strains from outpatients than in those from inpatients (P < 0.05). In conclusion, these data suggest that investigation for C. difficile infection should be taken into account in paediatric diarrhoea in both inpatients and outpatients in developing countries.

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.