To determine changes in suckling neonatal beef calf blood chemistry and body temperature during the first 72 h of life, jugular blood samples and rectal temperatures were obtained from 24 healthy, fall-born Angus-cross and Hereford calves (average calving date = September 11) at 0 (after standing, but pre suckling), 6, 12, 24, 48, and 72 h postnatally. Serum chemistry panels were conducted, and plasma triglycerides and serum non-esterified fatty acids (NEFA) were also determined. Data were analyzed using sampling hour as a repeated effect, and individual data points were compared to adult bovine reference intervals. All serum chemistry measures were affected by hour of age except bicarbonate (P = 0.48). Serum glucose, total protein, and globulin concentrations increased (P < 0.001) from 0 to 24 h. Plasma triglycerides increased (P < 0.001) from 0 to 6 h and 24 to 72 h. Serum urea nitrogen increased (P < 0.001) from 0 to 6 h and decreased (P = 0.01) from 48 to 72 h. Serum NEFA and creatine kinase (CK) increased (P < 0.001) from 0 to 6 h, but decreased (P < 0.02) from 12 to 72 h. Serum magnesium, aspartate aminotransferase (AST), and total and direct bilirubin increased (P < 0.001) during the first 12 h, then decreased (P < 0.001) from 24 to 72 h. Gamma-glutamyl transpeptidase (GGT) increased (P < 0.001) from 0 to 12, and then decreased (P < 0.003) from 12 to 48 h. Serum creatinine decreased (P < 0.001) from 0 to 72 h, and albumin decreased (P < 0.01) from 0 to 12 h but increased (P < 0.001) from 24 to 48 h. Anion gap decreased (P < 0.05) from 6 to 24 h. No serum components measured were within adult bovine reference intervals for all calves at all sampling times. All serum albumin concentrations were outside of reference intervals, and the majority of serum glucose, calcium, phosphorus, potassium, CK, GGT, and total bilirubin were outside of reference intervals. More serum chemistry measures diverged from reference intervals in the first 24 h of neonatal life. Rectal temperature decreased (P = 0.003) from 0 to 6 h of age, then increased (P < 0.02) from 12 to 48 h. In summary, blood chemistry profiles of healthy, suckling neonatal beef calves change over the first 72 h of life, indicating rapid changes in metabolism and physiology. Bovine reference intervals based on adults do not appear to represent neonatal calves; thus, neonatal status and animal age should be taken into consideration when interpreting serum chemistry in a clinical or research setting. (c) 2022 Elsevier Inc. All rights reserved.