Curtailing the observation of mice challenged with tetanus toxin in potency test of tetanus vaccine would reduce the agony of mice from spastic paralysis. From the viewpoint of animal welfare, we investigated the feasibility this measure. The potencies of 85 lots of vaccine obtained on the 4th day after challenge were compared with those obtained on the seventh day. No significant difference was found (P = 0.05), indicating that the observation period could be curtailed from 7 days to 4 days without impairing the assessment of the vaccine's potency.