After birth, as effectively monogastric animals, calves undergo substantial physiological changes to become ruminants by 3 months of age and reach sexual maturity at approximately 15 months of age. Herein, we assess longitudinal metabolomic changes in Holstein-Friesian (HF) heifers from birth until sexual maturity during this developmental process. Sera from 20 healthy, HF heifers were sampled biweekly from 2 weeks of age until 13 months of age and then monthly until 19 months of age. Sera were assessed using flow infusion electrospray high resolution mass spectrometry (FIE-HRMS) on a Q Exactive hybrid quadrupole-Orbitrap mass spectrometer for high throughput, sensitive, non-targeted metabolite fingerprinting. Partial least squares-discriminate analysis (PLS-DA) and unsupervised hierarchical clustering analysis (HCA) of the derived metabolomes indicated changes detectable in heifers’ sera over time. Time series analyses identified 30 metabolites that could be related to rumen development and weaning at ~ 3 months of age. Further time series analysis identified 40 metabolites that could be correlated to growth. These findings highlight the role of acetic acid and 3-PP within rumen development and growth, suggest that weaning induces elevated levels of fatty acyls in response to a post-weaning stress induced innate immune response and demonstrate the utilisation of fatty acyls in growth. The identified metabolites offer serum metabolites which could inform the nutrition and healthy development of heifers

Supplementary Data submitted as part of a submission to MDPI "Metabolites".