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The objectives of the present study were (1) to evaluate the effect of cholecalciferol treatment on Ca concentration during the first 10 DIM and (2) to evaluate the effect of treatment on subsequent health and performance. Multiparous Holstein cows (n = 375) were fed a negative DCAD diet (−31 mEq/kg DM) for the last 21 d of gestation. On d 275, the animals were randomly assigned to a control (CON) or a treatment group (VIT D). The latter ones received an injection of 12 × 106 IU cholecalciferol intramuscularly on the day of enrollment. Serum samples were taken on 1, 2, 3, 5, 7 and 10 DIM and analyzed for Ca, P, and Mg concentrations. In a subsample of cows (n = 100) concentrations of haptoglobin, NEFA, BHB and 25-OH vitamin D were determined on 1, 5 and 10 DIM. Repeated-measures ANOVA was performed to evaluate the effect of treatment on blood metabolites, rectal temperature and milk yield. Binary data such as retained placenta and clinical metritis were analyzed using logistic regression models. Cox proportional hazards were used to model time-to-event outcomes (i.e., time to pregnancy and culling). Results were considered significant when P < 0.05. Treatment increased serum concentrations of 25-OH vitamin D on d 1, 5 and 10 after calving. While Ca and P concentrations on 1 to 10 DIM were significantly increased in VIT D cows, Mg concentrations were significantly lower in comparison to CON cows. Interestingly, haptoglobin concentrations were significantly increased on 5 DIM in VIT D cows. In comparison to the CON group, VIT D cows showed elevated NEFA levels, a numerically higher incidence of retained fetal membranes and an increased the risk of incurring clinical metritis. In addition, treated cows produced 3.97 kg and 2.55 kg less energy-corrected milk on first and second test day, respectively, compared with CON cows. While there was no effect of treatment on culling until 300 DIM, time to pregnancy was delayed by 27 d in treated cows. In the present study, VIT D treatment had detrimental effects on health status and the subsequent lactation despite the beneficial effects on Ca homeostasis. Since this experiment was conducted on a single dairy farm, further research including more herds is warranted to evaluate whether the negative effects were herd specific.