Vehicle transportation represents acute stress to animals with release of catecholamines and glucocorticoids from the adrenal gland resulting in impaired metabolic state. Such changes in metabolism may be reduced by the application of suitable feed supplement. The aim of this study was to test the effects of lupin supplementation applied after 1-hour transportation. Ewes in the control group (n = 7) were fed on trefoil-grass silage and hay, while the diet of the experimental group (n = 7) was supplemented with lupin groats (Lupinus angustifolius, var. SONET; 500 g per head per day) for 8 days. In both groups, blood was collected on the day of transportation and on Days 6 and 11 thereafter. Total blood parameters were assayed using spectrophotometry and fractions of protein, cholesterol, and lactate dehydrogenase using agarose electrophoresis. Lupin increased the albumin: globulin (ALB : GLB) ratio and beta-hydroxybutyrate (BHB) concentration and reduced serum cholesterol and lactate, however it had no effect on body weight, body condition score (BCS), plasma glucose, serum protein, high-density lipoprotein (HDL), low-density lipoprotein (LDL), and very low-density lipoprotein (VLDL) cholesterol, lactate dehydrogenase (LDH) or alkaline phosphatise (ALP). Lupin may therefore be used as suitable feed supplement for sheep at times of high nutrient requirement.