Bacteroides fragilis strains with enterotoxic activity can be isolated from the faeces of newborn farm animals with diarrhoea and are called enterotoxigenic B. fragilis (ETBF). These strains can now be detected in an in-vitro cytotoxicity assay with HT-29 cells. In this study, 146 B. fragilis strains (95 faecal and 40 extra-intestinal isolates) and 64 Bacteroides isolates belonging to species other than B. fragilis were tested for their ability to produce enterotoxin. Sixteen strains of ETBF were identified; all belonged to the fragilis species and represented 11% of all B. fragilis examined. The prevalence was similar among extraintestinal and faecal strains, 11.5% and 10%, respectively. The production of enterotoxin in clinical isolates appeared to be associated with infections where tissue destruction was more prominent. Enterotoxigenicity was not associated with the presence of a plasmid and the plasmid profiles of ETBF strains that harboured plasmids were different. These results show that enterotoxin production by human isolates of B. fragilis is not uncommon and could represent a new virulence factor of B. fragilis.