Foot and Mouth Disease (FMD) is a highly contagious viral illness that predominantly affects cloven-hoofed animals. To identify and profile the predominant FMDV serotypes circulating in Baluchistan bovine and buffalo populations. Twenty tissue samples were collected and analyzed using sandwich indirect ELISA and RT-PCR to identify the presence and genetic characteristics of FMD virus. We identified 'O' serotype of FMD virus in all specimens. RT-PCR analysis yielded an impressive 90% positivity rate, confirming its diagnostic efficacy. While observations of LFBK cell line lead to the isolation of 18 distinct viruses with distinct Cytopathic Effects, two samples lacked detectable CPEs. Lower CT values, those observed in sample PK- 16 with CT of 15, indicated higher initial concentration of the target nucleic acid, indicating a potential for higher viral burden.The endemic prevalence of 'O' serotype in Balochistan necessitated targeted serotype-specific interventions, strengthened surveillance and strategic vaccination campaigns.