To shed light on the association of dietary fat with the development of colon cancer, we studied the ability of azoxymethane (AOM) to induce aberrant crypt foci (ACF) and biochemical changes in rats fed high- or normal-fat diets. Six-week-old male Fischer 344 rats were placed on a high-fat [7% (wt/wt) soybean oil + 15% (wt/wt) beef tallow] or a normal-fat (7% soybean oil, AIN-93G) diet. Rats fed each of these diets were given two weekly subcutaneous injections of AOM (15 mg/kg body wt) or saline at seven and eight weeks of age. Fecal samples were obtained at 10 weeks of age, and animals were sacrificed for ACF scoring and analysis of cecal contents at 13 weeks of age. We observed greater numbers of ACF in the high- than in the low-fat group. Biochemically, rats fed the high-fat diet showed dramatically elevated fecal and cecal long-chain free fatty acid levels and intestinal alkaline phosphatase activity. These animals also showed increased cholesterol and decreased coprostanol levels. We did not detect significant differences in the fecal and cecal concentrations of total and soluble bile acids or total neutral sterols (cholesterol + coprostanol) between the two groups. Thus a high-fat diet does show certain striking effects on colon biochemistry in rats.