The efficacy of 0.3% tacrolimus lotion (maximum dosage: 0.3 mg kg-1 per day) for treatment of atopic dermatitis (AD) was evaluated. Systemic absorption and effects on complete blood cell counts (CBC) and chemistry panels were also investigated. Eight dogs were assigned randomly to either a tacrolimus or a vehicle lotion treatment group. Both owners and investigator were blinded to the treatment. After 4 weeks, there was a 2-week wash-out period and treatments were reversed. Owners scored pruritus weekly while the investigator scored pruritus and erythema at the beginning and end of each treatment period. Investigator scores for pruritus in the tacrolimus group significantly decreased by the end of the study (P = 0.03). Investigator scores for erythema in the tacrolimus group were significantly lower than those in the placebo group at the end of the study (P = 0.005). There was no difference between groups with respect to owner scores for pruritus. No changes in the CBC and chemistry panels were noted. Mean blood concentrations of tacrolimus were below toxic levels.