The aim of this study was to determine the concentration of defensins BNBD‐1 and BNBD‐2 in fresh bovine milk and infant formula, and to compare the defensin concentrations of bovine milk and human milk. Abundant BNBD‐1 and BNBD‐2 was found in fresh bovine colostrum. However, compared to fresh bovine milk, BNBD‐1 and BNBD‐2 concentrations in infant formula were very low. The BNBDs concentration ranges were also less than β‐defensins in human milk. The BNBD‐2 concentration in fresh bovine milk was higher than that of BNBD‐1.