The development of antibody response against phospholipase A2 activity of Bothrops asper venom was studied in a group of adult and healthy horses used in the production of the polyvalent antivenom at the Instituto Clodomiro Picado. Simultaneously, the general condition of the animals during the immunization schedule was also studied. There was a great individual variability in the immune response, although most of the horses studied reached the highest neutralizing titer after injection of doses of venom of 30 mg and 50 mg. On the other hand, in horses that had been previously immunized and were infected for a second immunization, the highest antibody titer was observed 16-22 days after inoculation. There were only mild alterations in the general condition of the horses as a consequence of venom inoculation. However, local abscesses, fistulae and fibrosis were observed at the site of venom injection. On the basis of the conspicuous individual variability observed, it is proposed that the immune response in horses used in the production of antivenom must be evaluated on individual basis, instead of working with pools of serum samples.