The objective of this study was to determine variations of different physiological biomarkers in Clostridium perfringens (CP) infections in Balkhi breed of sheep in Khyber Pakhtunkhwa, Pakistan. Study was conducted on 184 sheep suspected of having enterotoxemia and 107 sheep were identified to be infected with Clostridium perfringens. Genotypic Analysis of all isolates from infected sheep was performed. Results showed, 53.27% isolates showed infection of CP type A, 10.28% of type B and 36.44% of type D. Animal infected with different serotypes (A, B, and C) were categorized into two groups healthy and diseased to compare the hematological and biochemical parameters. Hematobiochemical analysis indicated that mean erythrocyte count (RBC) and hemoglobin levels decreased while the mean leukocytes (WBC), platelets, packet cell volume (PCV) and total bilirubin levels increased significantly (P<0.05) in sheep infected with Clostridium perfringens type A. Sheep infected with Clostridium perfringens type B and type D showed significant (P<0.05) decreases in erythrocytes counts (RBC) and hemoglobin levels while the mean WBC, platelets, packet cell volume, serum creatinine, bilirubin, liver enzymes, total, glucose and urea in blood significantly (P<0.05) increased. Fluctuations in mean erythrocyte counts (RBC), hemoglobin, PCV and total bilirubin were beyond the limits while others were within normal ranges in sheep infected with Clostridium perfringens type A. The total bilirubin, Liver enzymes, creatinine level, glucose and urea levels in blood were abnormal while others were within normal ranges in sheep infected with CP type B and type D. by observing these changes it will be possible to diagnose disease at initial stages.