The aim of the study was to investigate the ability of vitamin C to increase the antioxidative and immunomodulating potential in healthy dogs. Fifteen dogs were tested for the effects of orally administered vitamin E (60 mg dl-alpha tocopheryl acetate) in combination with vitamin C (0, 30 or 60 mg ascorbic acid crystalline). Three treatments (0, 30, 60 mg vitamin C) were tested in a 3 x 3 cross-over study in three periods of 36 days. Pre-prandial blood samples were taken for analysis of vitamins C, E, A, retinyl palmitate and stearate, antioxidant status [Thiobarbituric acid reactive substances (TBARS) and uric acid], biochemical and haematological analysis. Subpopulations of lymphocytes, mitogen-induced peripheral blood mononuclear cell proliferation (PBMC) and serum IgA and IgG concentrations were determined. There was a trend (p = 0.056) for an increased plasma vitamin C concentration by vitamin C supplementation. There was no evidence that dietary treatment altered neither the other plasma vitamin concentrations nor TBARS and uric acid concentrations nor the subpopulations of the lymphocytes except for the number of CD4+ lymphocytes that increased with vitamin C supplementation. There was no effect of vitamin C on serum IgA and IgG concentration. A significant time x treatment interaction was demonstrated on PBMC's to pokeweed, with an increase observed by 30 mg vitamin C supplementation but a decrease by 60 mg vitamin C supplementation. There was no clear evidence for an effect of dietary vitamin C on antioxidative capacity in healthy dogs fed a diet with vitamin E concentrations well above the recommendations. Yet, a limited number of immunological parameters were slightly affected.