Buffalopox (BPX) is a zoonotic infection affecting humans, cattle and buffalo. Though, the infection is not associated with heavy mortality even then it adversely affects the working capacity and production potential of the animals resulting huge losses in terms of economy. In present study blood (n=163) and scab (n=19) samples were collected from healthy and BPX suspected buffalo (n=975). Blood samples were used to determine red blood cell (RBC) count, hemoglobin (Hb) concentration, packed cell volume (PCV), platelet (PLT) count, total leukocyte count (TLC) and differential leukocyte count (DLC) in apparently healthy and diseased animals. The data on hematological parameters showed non-significant (P > 0.05) difference on RBC count, Hb concentration, PCV and PLT in diseased and healthy animals. Significant (P < 0.05) increase in TLC was observed in diseased buffaloes due to increase in number of lymphocytes regardless of age groups of animals. Scab samples were injected intradermally in healthy rabbits for BPXV confirmation. Rise in body temperature and development of pit-forming lesions with a specific pattern was suggestive of BPX infection. Histopathologic observation of skin samples of rabbits revealed marked hyperkeratosis, ballooning degeneration and large intracytoplasmic-eosinophilic inclusion bodies in keratinocytes and mononuclear cell infiltration in epidermis and dermis.