Background: Transition period is a critical period in which most dairy animals undergo a period of negative energy balance (NEB), hypocalcemia, insulin resistance, hypoglycemia and susceptibility to infectious diseases. The critical changes in biochemical indicators are well reported in dairy cows whereas; in dairy buffaloes limited data related to alterations in hemato-biochemical changes have been reported so far. The current study aimed to evaluate the hemato-biochemical changes in buffaloes during transition period. Methods: A total number of 210 buffaloes aged between 3 to 14 years with parity ranged from 1st to 4th, divided in three groups (n=70; each) during transition period were used for the study. Three groups viz Group I (-30 days), Group II (0 days) and Group III (+30 days) were subjected for evaluation of hemato-biochemical parameters. Result: Our study showed significant increase (p less than 0.05) in WBC (103/µl), lymphocytes and granulocytes count at the time of calving. Significant decrease (p less than 0.05) in the level of RBC, Hb and HCT were seen in Group-I and II. In biochemical analysis, parameters like albumin, total protein, triglyceride and A:G ratio showed significant (p less than 0.05) decrease from Group-I to II. Calcium and Ca:P ratio decreased significantly (p less than 0.05) along with suppressed levels of sodium and potassium levels in Group-III. Thus, the present study showed significance of hemato-biochemical analysis during transition period for possibility of occurrence of production diseases.