Concomitantly to the negative energy balance in early lactation, low total cholesterol (TC) concentrations are observed. We have investigated if the level of TC in early lactation is related to short-term adaptations of metabolism and milk production, when cows are exposed to a transient concentrate withdrawal of one week that further aggravates energy deficiency. Multiparous Holstein cows (n=15) were investigated during a period of 21 days beginning at 24±7 DIM (mean±SD). Cows were kept on pasture and received additional concentrate in experimental week 1 and 3, while in week 2 concentrate was withdrawn. Blood was sampled once and milk samples twice a day. Based on their average TC concentration during week 1 (prior to concentrate withdrawal), cows were retrospectively grouped into a high (HC; n=8, TC ≥3.36 mmol/l) and a low TC group (LC; n=7, TC <3.36 mmol/l). A mixed model with group, time as fixed and cow as random effect was used to evaluate metabolic and performance responses in terms of concentrate withdrawal. Concentrations of phospholipids and lipoproteins were higher in HC compared to LC throughout the study (P<0.05). During concentrate withdrawal in week 2, milk yield and concentrations of glucose and insulin decreased similarly in both groups, while milk fat and acetone content, and plasma concentration of BHB increased (P<0.05). Compared to initial values, plasma NEFA, TG, VLDL increased in both groups within 2 days after concentrate withdrawal (P<0.05), but declined again thereafter. Changes in insulin and glucose occurred within 1 day. Despite re-introduction of concentrate in week 3, milk yield in HC was lower compared to week 1 (P<0.05) and not different between weeks 2 and 3 (P>0.05), whereas milk yield in LC fully recovered. Activity of aspartate aminotransferase was higher in HC compared to LC in week 2 (P<0.05). In conclusion, circulating cholesterol concentrations are associated with the extent of short-term adaptation responses to energy availability in early lactation.