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Metabolization of dietary soy isoflavones has been associated with favorable prostate cancer risk reduction. However, the metabolites that are effective chemopreventive agents have yet to be identified. Due to the large variability in gut microbial equol (Eq) production among humans, a key question remains as to whether dietary phytoestrogens (e.g. soy isoflavones) are associated with Eq production in the gut. In this study, the formation of Eq by human gut microbiota was determined. Soy isoflavone and genistein concentrations were measured in human feces taken from men consuming a low-soy diet and at 12-week intervals during a 30-month study of soy intake. A 3-day human fecal collection was performed at 3 months following soy isoflavone and genistein supplements. Cultures were prepared to measure Eq formation and bacterial numbers and Eq concentrations were determined. In subjects who consumed soy isoflavone and genistein supplements, Eq was detected in feces at day 3, 8, and 12 of the study. Increasing amounts of Eq were associated with greater amounts of genistein consumed, suggesting that there was an association between genistein and Eq concentrations. The mean Eq concentration in the presence of genistein was significantly higher than in the absence of genistein at day 3 (3.87 ± 0.72 vs