Normal bacteria can translocate (migrate) across an intact intestinal mucosa, but the anatomical site of bacterial translocation has not been defined. Gastrointestinal (GI) cross-sections were obtained from mice monoassociated with high cecal concentrations of Escherichia coli, Proteus mirabilis, or Enterococcus faecalis. As previously reported (C. L. Wells, R. P. Jechorek, and K. J. Gillingham, Arch. Surg. 126:247-252, 1991), these mice had viable translocating bacteria recovered from mesenteric lymph nodes and livers. No abnormal GI histology was noted. Immunofluorescence was used to localize GI bacteria, and similar observations were made with each of the three bacterial species. Smaller numbers (P < 0.01) of bacteria were observed in stomach and small intestinal tissues than in cecal and colonic tissues, suggesting that the preferred site of tissue penetration for intestinal E. coli, P. mirabilis, and E. faecalis might be the cecum or colon as opposed to the stomach or small intestine.