ABSTRACT: The objective of this experiment was to examine the effect of milk replacer (MR) feeding rate (FR) and frequency (FF) on glucose metabolism before and after weaning during summer and winter in the subtropical climate of the southeastern United States. Holstein calves (n = 48/season) were enrolled at 8 d of age (DOA) in the summer (June to August, body weight = 40.6 ± 0.7 kg) and winter (November to January, body weight = 41.9 ± 0.8 kg). In each season, calves were randomly assigned to 1 of 4 treatments in a 2 × 2 factorial arrangement including 2 FR [0.65 (low) or 0.76 kg of solids/d (high) of a 26% CP and 17% fat MR] and 2 FF [2× (0700 and 1600 h) or 3× (0700, 1600, and 2200 h)]. Calves were managed similarly and housed in polyethylene hutches bedded with sand. Milk replacer (12.5%) was fed based on treatments until 42 DOA when FR was reduced by half and offered 1×/d (0700 h) for 7 d. Plasma was collected weekly at 1400 h for analyses of glucose and insulin concentrations in all calves. Pre- and postprandial glucose and insulin concentrations of a subset of calves (n = 10/treatment per season) were measured on 20 DOA. A subset of calves (n = 8/treatment per season) was subjected to an intravenous glucose tolerance test (GTT) on 27 and 57 DOA and insulin challenge on 28 and 58 DOA at 1030 h. Average ambient temperature was 26.1 ± 2.2°C in summer and 12.9 ± 5.4°C in winter. During the preweaning period in both seasons, feeding high increased plasma glucose concentrations compared with low, and increasing FF reduced basal insulin concentrations. Compared with 2×, feeding 3× did not affect postprandial glucose but lowered insulin in the summer, whereas in the winter, increased glucose from 30 to 180 min but lowered insulin from 240 to 420 min after MR feeding. Following GTT before weaning in both seasons, 3× reduced insulin increment and area under the curve compared with 2× without affecting glucose disposal. After weaning, treatment did not affect glucose disposal or insulin responses after GTT during winter, but calves fed 3× had faster glucose disposal and stronger insulin responses than 2× during summer. In both summer and winter, preweaned calves fed 3× had greater decrement and area under the curve of plasma glucose after insulin challenge, suggesting enhanced peripheral tissue insulin response compared with 2×. This effect persisted after weaning only during summer. Increasing FR had no effect on metabolic responses in both seasons. In conclusion, increasing MR FF from 2 to 3 times per day reduced insulin secretion but enhanced insulin response on peripheral tissues of preweaned calves regardless of season.