Eighteen Beagle dogs were used to evaluate the effects of bovine lactoferrin (bLF) on immune function and faecal microbial populations. The study comprised three feeding periods, each lasting four weeks. After an initial control Period 1, six dogs per group were supplemented with 0, 120 and 1800 mg bLF/kg dry diet, respectively (Period 2). In Period 3 dogs received again control diets. Peripheral blood mononuclear cell subsets, lymphocyte proliferative response to concanavalin A, phytohaemagglutinin and pokeweed mitogen and plasma IgA and IgG concentrations were analysed. The faecal concentrations of aerobic and anaerobic bacteria, Escherichia coli, Clostridium perfringens, Lactobacillus spp. and Bifidobacterium spp. were determined by cultural methods. Supplementation of bLF increased the number of monocytes, T cells and cytotoxic T cells in the blood and the proliferative response of peripheral blood mononuclear cells. The leukocyte counts were not affected, except monocytes that increased after the supplementation with bLF. Plasma immunoglobulin concentrations were unchanged by treatment. Dogs supplemented with bLF tended to have lower faecal concentrations of E. coli and Clostridium perfringens. In conclusion, bLF seems to alter indices of the cellular immune response and faecal microbial populations of healthy adult dogs.