The primary objective of current study was to evaluate the effects of different anastrozole dosages on the physiological performance, hematological profile, and serum biochemical parameters of broiler roosters. A total of Twenty-six Lohmann Brown roosters were randomly assigned to four treatment groups. The first group (T1) served as the control and received no anastrozole, while the other groups (T2, T3, and T4) were administered 0.2 mg, 0.4 mg, and 0.6 mg of anastrozole per day, respectively. The first and second groups consisted of six birds each, while the third and fourth groups had seven. The results demonstrated a significant improvement (P ≤ 0.05) in several physiological and biochemical parameters in the group receiving 0.6 mg of anastrozole (T4) compared to the control. Notably, hematological analyses revealed a significant increase in red blood cell (RBC) count, hemoglobin (Hb) concentration, and packed cell volume (PCV), indicating improved oxygen transport and erythropoietic activity. Additionally, total protein and globulin concentrations in serum were significantly elevated (P ≤ 0.05), suggesting enhanced protein metabolism and immune response. Moreover, a significant reduction (P ≤ 0.05) in serum lipid profile, including total cholesterol, was observed, reflecting improved lipid metabolism. Importantly, glucose concentration remained within the normal physiological range. In conclusion, the administration of 0.6 mg of anastrozole demonstrated beneficial effects on the physiological performance of broiler roosters by enhancing hematological parameters and optimizing metabolic processes, suggesting its potential role in improving poultry production efficiency.