Maternal investment plays a critical role in neonate survival, and adults can improve survival of offspring by defending them against predators. However, limited information exists documenting ungulate aggression toward humans in defense of neonates. During captures of neonates in spring 2007 and 2008 in north-central South Dakota, we documented 24 aggressive encounters by adult female and yearling male and female white-tailed deer (Odocoileus virginianus) defending neonates. Eleven (45.8%) aggressive encounters included yearlings accompanying adult females. Mean ages and weights of neonates that were aggressively defended were greater (P < 0.0001) than ages and weights of those that were not; adults began protecting neonates at approximately 4 days of age. Male fawns were more likely (P  =  0.013) to be defended than female fawns. Examination of our data suggests that sex- and age-biased maternal defensive behavior exists in white-tailed deer, and that deer biased maternal investment toward older, male neonates.