The purpose of this trial was to examine the potential of a new Escherichia (E) coli J-5 mutant strain bacterin to reduce the severity of clinical disease caused by subcutaneous challenge with endotoxins of Gram-negative bacteria in calves. Day-old to 3-day old calves (n = 40 per study phase) were randomly assigned to either of two treatment groups, i.e. a vaccinated or a placebo group. Calves in the vaccinated group received an inactivated bacterin containing a J-5 mutant strain of E. coli via subcutaneous route at 2-4 days of age and at 14 days thereafter. The placebo contained only adjuvant and saline in lieu of the antigen. Lipopolysaccharides (LPS) originating from E. coli were administered subcutaneously 3 weeks after the booster dose. The LPS challenge dosages were 1 and 8 microg/kg in study phases I and II, respectively. Various clinical, physiological, hematological, and serological parameters were measured at specific time intervals after challenge. The data were mostly analysed using peak changes from baseline recorded during the observation period. By the time of challenge the titers in vaccinated calves had increased significantly more than in the unvaccinated controls. Disease severity following subcutaneous challenge was dose dependent. In phase I, placebo calves were only mildly challenged whereas in phase II placebo calves showed a moderate challenge. After a mild challenge, there was little evidence of protection due to vaccination as only attitude was significantly improved in the vaccinates. In contrast, after a moderate challenge rectal temperature, hematocrit, blood glucose concentrations, and leukocyte changes were significantly better in the vaccinated group. In conclusion, the results of this study show that following a subcutaneous endotoxin challenge that induces a moderate clinical response, calves that were previously vaccinated with the E. coli J-5 bacterin were better protected than those in the placebo group.