Actinomycetes are fascinating group of microorganisams. Actinomycetes are Gram positive having G + C (>55%) content in their DNA. The aim of present study is to screen potential strain of Actinomyctes and checking against known Gram positive and Gram negative pathogenic bacteria by using cup plate method. The isolates( VPD 14 and VPD 19) could not show zone of inhibition against Gram positive bacteria but the same strains were considerably significant against Gram negative bacteria (E-coli and Pseudomonas) showing zone of inhibition 0.2cm,0.4cm,0.1cm and 0.3cm respectively.