This study was conducted to determine seroprevalence of Brucella infection in cows in Kafer El-Sheikh and Qualyobia governorates and evaluate the biochemical parameters of infected cows that reflect the effects of Brucella infection on animal health and performance. A total of 240 blood samples were collected from cows (120 from kafr- El Sheich and 120 from Qualyobia). All sera were tested by Rose Bengal Plate Test (RBPT), Buffered acidified Plate Antigen Test (BAPAT) and Complement Fixation test (CFT). Brucellosis antibodies were detected in 18 of 120 (15%) and 6 of 120 (5%) cattle serum samples collected from Kafr-ElSheich and Qualyobia governorates, respectively. There was a significant correlation between brucellosis antibodies in cattle serum samples and location where the blood samples be collected. The highest prevalence of brucellosis antibodies was detected in cattle serum samples > 6 years of age in two governorates. Biochemical examinations revealed significant increases in Alanine aminotransferase (ALT), Aspartate aminotransferase (AST), Creatinine kinase (CK) activities and cholesterol level in serum of Brucella infected cows in comparison with healthy cows. Meanwhile, non significant changes were reported in urea, creatinine, glucose and C-reactive protein (CRP) levels. Changes in biochemical parameters reveled that brucellosis has harmful effects on cow’s health. It could be concluded that determination of biochemical parameters in cow can determine the extent of damage caused by Brucella infection.