To shed light on the association of intestinal microflora with the development of colon cancer, we studied the modifying effects of intestinal microflora on the occurrence of 1,2-dimethylhydrazine (DMH)-induced colonic aberrant crypt foci (ACF) in germfree (GF), gnotobiotic (GB) and conventionalized (Cvd) rats. In the first part of this study, 10 week old germfree Fischer-344 rats were randomly assigned to three groups and two groups of rats were orally inoculated with mixtures of pure culture of Escherichia coli, Enterococcus faecium, and several strains of Bacteroides and Clostridium species (GB), or feces from conventional rats (Cvd). Inoculated rats were given two weekly i.p. injections of DMH (20 mg/kg body wt) at 13 and 14 weeks of age. Rats were sacrificed 11 or 34 weeks after the last DMH injection for ACF scoring. The total number of ACF, ACF with four or more crypts/focus, and mean number of aberrant crypts per focus (crypt multiplicity) in GB rats sacrificed at week 34 were 168% (P < 0.001), 442% (P < 0.001) and 138% (P < 0.001) of those in GF rats, respectively. On the other hand, the same values in Cvd rats were 42% (P < 0.001), 147% (P = 0.246) and 159% (P < 0.001) of those in GF rats, respectively. Similar results were observed in rats that were sacrificed at week 11. In the second part of this study, the effect of colonization of Bifidobacterium breve on the ACF profiles was examined in GB rats. The number of ACF with four or more crypts/focus and crypt multiplicity in GB plus B. breve rats at week 11 were significantly lower than those of GB rats (P < 0.01, and P < 0.05, respectively), although the former was not statistically significant at week 34. These findings suggest that some intestinal bacteria might behave as promoters and some as anti-promoters in colon carcinogenesis.