Bovine colostrum is important for neonates' health due to its nutritive and nonnutritive components. Heat treatment of colostrum is a well-established management tool, but it may influence colostrum components and affect the health status of calves. In our previous studies, we had shown that colostrum proteome and serum proteome of calves were altered by heat treatment to different degrees. Our objectives in this study were to investigate the effects of heat treatment on colostrum metabolome and the effect of feeding heat-treated colostrum on the serum metabolome of newborn calves. Further, the changes in serum metabolome from before to after colostrum feeding were characterized. Newborn Holstein female calves (n = 10) were randomized within pairs and fed heat-treated (n = 5; 60 °C, 60 min) or raw (n = 5) colostrum at 8.5% of birth BW by esophageal feeder within 1 h of birth. After a single colostrum feeding, calves were not fed until after the 8 h time point.