Abstract: Buffalopox is one of the zoonotic infections having a significant public health impact. The virus is closely related to Vaccinia virus. There is no prophylactic available to combat the buffalopox infection. Ample number of reports is available indicating the use of herbal preparations for veterinary use in India. In this resesrch, four plants having known medicinal importance were screened for buffalopox virus inhibition in vitro. Of the four plant tested, extract from Eugenia jambolana leaves had an inhibition of 98.52% at its maximum non toxic concentration (1999.73�0.50 �g mL 1) in all cytopathic effect inhibition assays. The inhibition of buffalopox virus replication was further confirmed using PCR (ATI and C18Lgenes) and Real Time-PCR (C18L gene) assays specific for buffalopox virus. Our results indicate that the extract from Eugenia jambolana leaves inhibit the buffalopox virus in vitro.