The hypothesis was tested that the maternal supply with essential fatty acids (EFA), especially α-linolenic acid (ALA), and conjugated linoleic acid (CLA) affects glucose metabolism in neonatal calves. Dairy cows were fed corn silage based TMR with low fat and ALA content for the last 3 mo of gestation. All calves were delivered by dams abomasally supplemented either with coconut oil (76 g/d; CTRL; n = 9), linseed and safflower oil (78 + 4 g/d; EFA; n = 9), Lutalin (10 g/d c9, t11 and t10, c12 CLA, resp.; CLA; n = 9) or a combination of EFA and CLA (EFA+CLA; n = 11) during the last 9 wk of gestation and lactation. Each calf was fed with colostrum from its own dam. Plasma concentrations of glucose and insulin were measured daily in calves from d 1 until d 5 of age before feeding. On d 4, calves were fed their morning meal (6% of BW) mixed with 10 mg/kg BW [¹³C₆]-glucose and 0.5 g/kg BW xylose. Immediately afterward, an intravenous bolus dose of [6,6-²H₂]-glucose (5 mg/kg BW) was applied. Blood samples were taken to measure plasma ¹³C and ²H glucose enrichments and to calculate rates of glucose appearance and fractional first pass splanchnic uptake (FPU) of glucose. Data analysis was conducted by means of the MIXED procedure of SAS containing treatment, time, treatment × time, and sex as fixed effects and supplementation period and duration of gestation as covariates. Before first colostrum intake, plasma glucose concentration was higher (P < 0.05) in EFA than in CTRL and CLA calves. Plasma insulin concentration on d 4 was higher 60 min after feeding in EFA than in CLA (P < 0.05) and 120 min after feeding in EFA+CLA than in CTRL (P < 0.05). Plasma xylose and [¹³C₆]-glucose enrichment increased (P < 0.05) in all groups after feeding, the increase of [¹³C₆]-glucose was faster in EFA+CLA than in CLA, and the decrease of xylose was faster (P < 0.05) in EFA than in CTRL. Glucose FPU did not differ among treatment groups. Results indicate minor changes of postnatal glucose metabolism with respect to maternal fatty acid supply, but point at an effect of maternal EFA on fetal glucose supply.