This study examined developmental differences in infants' salivary alpha-amylase (sAA) and cortisol levels and responses to the well-baby exam/inoculation stress protocol at 2, 6, 12, and 24 months of age. Mother-infant pairs (n=85; 45 girls) were assessed during well-baby visits and saliva was sampled before the well-baby exam/inoculation procedure (pre-test) and at 5, 10, and 20 min post-inoculation stress. Older infants (24 months) had higher levels of sAA than younger infants (2, 6 and 12 months). Stress-related sAA increases were evident at 6 and 12 months, but not at 2 or 24 months of age. Stress-related cortisol increases were present at 2 and 6 months, but not at older ages. Mothers had higher sAA levels than their infants, but did not show sAA or cortisol increases to their infants' inoculation. Pre-test, maternal and infant sAA levels were positively correlated (rs .47 to .65) at 6, 12, and 24 months of age, but not at 2 months. These findings suggest that the association between the sympathetic branch of the autonomic nervous system and the secretion of sAA develops between 2 and 6 months of age, when levels of sAA are responsive to exposure to a painful stressor.