A small percentage of mature mares continue to exhibit estrous cyclicity during the non-breeding season which is of interest because of the importance of timing of the breeding season to the equine breeding industry. Previously, it was demonstrated that the continuation of estrous cycles was more likely to occur in mature than young mares. Additionally, an apparent association exists between elevated body fat and increased circulating concentrations of leptin, and the occurrence of estrous cycles during the non-breeding season. Two experiments were conducted to test the hypothesis that pharmacological manipulation of body fat and leptin by administration of the beta(2)-adrenergic receptor agonist clenbuterol, with and without continuous treatment with melatonin, would increase the proportion of mares entering anestrus or advance its timing. In Experiment 1, eight mature mares were administered clenbuterol (3.2 microg/kg) daily from October through January and six mares were untreated. In Experiment 2, eight mares were administered clenbuterol daily from June through December, eight mares were treated with continuous release melatonin implants, and seven mares were treated with both clenbuterol and melatonin. Melatonin treatment was included to confirm previous results that constant treatment with melatonin did not advance the timing of anestrus. In both studies, leptin concentrations (P<0.01, both experiments) and body fat percentage (P<0.01, Experiment 1; P<0.05, Experiment 2) declined in response to daily clenbuterol treatment. Unexpectedly, however, this decrease was attenuated in mares also treated with melatonin (P<0.05). Although treatment with clenbuterol was associated with a decline in body fat and circulating concentrations of leptin, the timing or proportion of mares exhibiting anestrus was not modified (P>0.05). The results demonstrate the manipulation of body condition via pharmacological intervention does not modify the mechanisms controlling seasonal anestrus in the mare.