The action of bile on bifidobacteria was studied by measuring changes in zeta potential, hydrophobicity and adherence to enterocyte-like cells in vitro. Highly hydrophobic strains shocked with bile displayed more negative zeta potential values and a decrease in adherence. When a non-hydrophobic non-adherent strain (CIDCA 5324) was shocked with bile, an increase in hydrophobicity was observed. However, no changes of zeta potential or adherence properties were apparent. The action of the bile components was different from the action of whole bile. Cholate and deoxycholate produced a decrease in the negativity of zeta potential values of all strains studied whereas taurocholate displayed a shift in zeta potential of hydrophobic strains to more negative values, thus explaining the decrease in the autoaggregation by charge repulsion. However, the decrease in zeta potential caused by cholate and deoxycholate did not increase autoaggregation in a hydrophobic non-adherent strain (CIDCA 531). This suggests that other forces are contributing to autoaggregation.