Isolates of Clostridium perfringens recovered from Costa Rican patients with antibiotic-associated diarrhoea are mostly enterotoxin-negative and susceptible to first-choice antimicrobials

Abstract

To assess the prevalence of enterotoxigenic Clostridium perfringens among adults suffering from antibiotic-associated diarrhoea in a Costa Rican hospital, faecal samples were analysed from 104 patients by a cultivation approach. The 29 strains obtained, which accounted for an isolation frequency of 28 %, were genotyped and investigated with regard to their in vitro susceptibility to penicillin, imipenem, cefotaxime, chloramphenicol and metronidazole using an agar-dilution method. A multiplex PCR for detection of the toxins α, β and ϵ predictably classified all faecal isolates as biotype A. An agglutination assay revealed that only one isolate synthesized detectable amounts of enterotoxin (detection rate 3 %). This result was confirmed by a PCR targeting the cpe gene. The spores of the only CPE+ isolate did not germinate after incubation for 30 min at temperatures above 80 °C. Most isolates were susceptible to first-choice antimicrobials. However, unusual MICs for penicillin (16 μg ml−1) and metronidazole (512 μg ml−1) were detected in one and three isolates, respectively. The low incidence of enterotoxigenic strains suggests that C. perfringens was not a major primary cause of antibiotic-associated diarrhoea in this hospital during the sampling period.