Bacillus cytotoxicus, a member of the Bacillus cereus group, is a thermotolerant species originally reported from a lethal foodborne infection in France in 1998. The strain NVH391-98, isolated from this outbreak, produces cytotoxin K1, a potential cytotoxic enterotoxin. However, the habitat and diversity of B. cytotoxicus isolates so far have been poorly explored. The objective of this study was to assess the prevalence of this bacterium in different food products (mainly dried) and to estimate its diversity. Among the 210 samples analyzed, all potato flakes contained the bacterium at low concentrations (≤102 CFU/g). However, prepared and kept at room temperature for 2 days, the puree contained ca. 105 CFU/g B. cytotoxicus. Besides potato flakes, some samples of millet flour, salted potato chips, and soups also contained B. cytotoxicus. From these samples, 55 thermotolerant B. cytotoxicus isolates were obtained. When classified into six distinct random amplified polymorphism DNA patterns, they showed the existence of 11 distinct plasmid profiles. Although most isolates (including the reference strains NVH391-98 and NVH883-00) contained no detectable plasmid, some displayed one to three plasmids with sizes from ca. 8 to 90 kb. It also emerged from this study that a single food sample could contain B. cytotoxicus isolates with different genetic profiles. HIGHLIGHTS