The present study aimed to evaluate the hematological and serum biochemical parameters in buffaloes and cows infected with foot and mouth disease (FMD) virus. Forty buffaloes and cows (6 months - 2 years old) were used. Twenty buffaloes and cows clinically healthy and were used as control, the other twenty buffaloes and cows showed characteristic clinical signs of FMD. Blood samples were collected and serum was separated from both groups to measure the hematological and biochemical parameters. Infected buffaloes and cows showed fever, anroxia, excessive foamy salivation and ulcer formation in the mouth. Compared to the control buffaloes, erythrocytic count (RBCs), hemoglobin (Hb) and packed cell volume (PCV) in the FMD infected buffaloes were significantly decreased, while mean corpascular hemoglobin (MCH) was significantly increased. In addition, significant increase in toltal leukocyte count (TLC), neutrophils, with a significant decrease in eosinophil and lymphocyte counts were observed.Moreover, cows infected with FMD virus showed no significant changes in the erythrogram, while, significant increases in TLC and neutrophils, with a significant decrease in eosinophil and lymphocyte counts were observed when compared with the control group. There were significant decreases in the levels of serum total proteins, albumin and globulin, while serum asprtate transferase (AST), creatinine phospho-kinase (CPK), glucose and cortisol levels were significantly increased in the infected animals. Tissue and saliva specimens were collected and analyzed by using universal primer using reverse-transcription polymerase chain reaction (RT-PCR). All the identified viruses were of serotype ‘O'. It is concluded that FMDV serotype ‘O' is circulating among cows and buffaloes in the study area.