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The use of oral rehydration solutions (ORS) is well established as an effective treatment to correct water-, electrolyte-, and acid-base balance in diarrheic calves. The main ingredients of a commercial ORS are Na, glucose, and alkalinizing agents, such as NaHCO3. Particular importance is attributed to the combination of glucose and Na at a specific ratio to optimize intestinal sodium, and thereby water uptake, through the sodium-glucose co-transport. Enhancing intestinal Na absorption by combining glucose and Na in an ORS has the potential to improve the alkalinizing effect of an ORS according the strong ion theory. The objective of this study was to investigate the effect of glucose on the alkalinizing effect of NaHCO3 when administered orally. Nine healthy neonatal Holstein-Friesian calves underwent 3 oral treatments with 2-L solutions of NaHCO3 (150 mmol/L), glucose (300 mmol/L), and glucose + NaHCO3 (300 mmol/L + 150 mmol/L, respectively) in randomized order. Arterial and venous blood was obtained before treatment and in 30-min intervals thereafter for blood gas analysis and determination of plasma protein and electrolyte concentrations. Urine was collected volumetrically to determine urine volume, osmolality, pH, net acid excretion, and renal Na excretion after treatment. Plasma volume changes were extrapolated from plasma protein concentration changes. Treatment and time effects were tested with repeated measures ANOVA. Only subtle differences between oral administration of NaHCO3, with and without glucose, were observed for the change of the standard HCO3 concentration relative to baseline. No differences in plasma Na, plasma volume expansion, renal Na, net base excretion, urine volume, or pH could be identified between animals treated orally with NaHCO3 with and without glucose. Similarly, no differences in blood glucose concentration, plasma volume expansion, urine volume, or renal glucose excretion were observed in the 8 h after treatment when comparing oral glucose treatment with and without NaHCO3. Our results indicate that combination of NaHCO3 with glucose in a hypertonic ORS only had a minor effect on the alkalinizing effect of NaHCO3, which is unlikely to be of clinical relevance. The combination of NaHCO3 and glucose neither improved Na, glucose, nor water absorption in euhydrated neonatal dairy calves, questioning the relevance of a specific ratio between Na and glucose in ORS for calves.