A Salmonella typhimurium strain harboring stable mutations in both ompC and ompF was constructed from the mouse-virulent strain S. typhimurium SL1344. When administered orally to BALB/c mice the strain was attenuated, with the 50% lethal dose (LD50) reduced by approximately 1,000-fold. However, the intravenous LD50 was reduced only by approximately 10-fold. The ompC ompF mutant persisted in murine tissues for several weeks following oral challenge, and mice immunized with this mutant were well protected against challenge with virulent SL1344. A strain harboring a stable mutation in tppB behaved in a manner similar to that of strain SL1344 in vivo, while a strain harboring mutations in ompC, ompF, and tppB behaved as an ompC ompF mutant in vivo, indicating that the tppB operon is not required for virulence in S. typhimurium.