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Foot and mouth disease is among the top listed livestock diseases causing severe economic losses. The aim of the present study was to determine the seroprevalence of FMD and detection of FMD virus shedding in milk of apparently healthy buffaloes and cattle of Pakistan. A total of 30 dairy farms were selected and registered in rural areas of Punjab consisting of minimum 15 animals. A total of 180 serum samples were collected and subjected to Non structural proteins (NSP) ELISA. The milk of sero-positive animals were collected and detected for the presence of FMD virus using reverse transcriptase PCR (RT-PCR). The results of current study showed overall seroprevalence 71.66% (129/180) with 65.38% in buffaloes and 76.47% in cattle. The FMD virus was detected in 24.03% (31/129) of sero positive samples. Among the FMD virus positive samples 65.51% belongs to serotype “O” and 35.48% belongs to Asia I, while none of the other serotypes were detected. The detection of FMD virus from the milk of apparently healthy buffaloes and cattle is an alarming situation and it may be considered as a potential role in the transmission of FMD.