Abstract: Background: Oxidative stress biomarkers and lipid profiles were used successfully as prognostic and diagnostic biomarkers of many animal diseases. However, their use in the diagnosis of ketosis in dairy cows at post-paturient period is not completely elucidated. Materials and Methods: Therefore, 25 cows suffered from ketosis at post-paturient period were used in the current study together with 20 healthy cows who served as a control. Blood samples were collected from diseased and healthy animals and the harvested serum were used for determination of oxidative stress biomarkers and the profiles of lipids, protein and enzymes. Results: The obtained results declared that, there was a significant (p≤0.05) increase in the levels of aspartate aminotransferase (AST), gamma glutamyl transferase (GGT), non-esterified free fatty acids (NEFA), β-hydroxylbutyric acids (BHBA), malonaldehyde (MDA) and nitric oxide (NO) in dairy cows affected with ketosis compared to control. Conversely, a significant (p≤0.05) decrease in the levels of glucose, total cholesterol, cholesterol ester, free cholesterol, triacylglycerol (TAG), superoxide dismutase (SOD) and reduced glutathione (GSH) were detected in diseased cows compared to control. Serum BHBA, NEFA, MDA and NO levels were positively correlated with each other’s and inversely correlated with activity of SOD and GSH concentration in cows affected with ketosis. Conclusion: Oxidative stress biomarkers and lipid profiles could be used as promising biomarkers for ketosis in dairy cows at post-paturient period. The antioxidant therapy may useful in the treatment of ketosis in cows at post-paturient period.