There is increasing interest in the use of natural antioxidant supplements in poultry diets as protection against the adverse effects of heat stress. The potential protective effect of boldo (Peumus boldus molina) leaf extract, which have antioxidant activity, were investigated against the harmful effects of heat stress in two broiler strains. Arbor Acres (AA) and Avian-48 (AV) chicks were divided into thermoneutral (TN) and heat stress (HS) groups and treated with 1 g boldo leaf extract/4 L drinking water during the heat stress period. HS reduced growth performance in both strains. The phagocytic index, phagocytic activity, and eosinophil and lymphocytes counts were significantly elevated in TN and HS AV birds but not altered in AA birds. Boldo extract treatment partially eliminated the previous negative impacts of heat stress. AA chicks were better able to withstand HS than AV chicks. Serum concentrations of total lipids and cholesterol were reduced in HS birds of both strains. Malondialdehyde, superoxide dismutase, and glutathione peroxidase levels were elevated but restored with the administration of boldo leaf extract in HS birds of both strains. Economic parameters were negatively affected by HS but restored to values close to those of the control group in boldo-treated HS birds. In conclusion, the administration of boldo leaf extract in drinking water was effective in neutralizing the harmful effects of heat stress on growth performance, blood indices, and economic parameters and improved the antioxidant defense system in heat-stressed birds.