Antagonistic microorganisms have been used as biological control agents to reduce the use of chemical fungicides in the control of crop diseases. The present work was conducted to determine the antagonistic potential of Bacillus sp. isolates against the soil-borne fungus Cochliobolus sativus, the causal agent of barley common root (CRR) disease. Out of 525 isolates, 40 were showed in vitro antagonistic activity against the virulent C. sativus isolate CRR16. On the basis of 16S rRNA gene sequencing Bacillus sp. isolates are identified as B. atrophaeus, B. subtilis, Paenibacillus polymyxa, B. amyloliquefaciens, B. simplex and B. tequilensis. Results showed that Bacillus sp. had significant (P<0.05) antagonistic activities against the C. sativus where the percentage of radial growth inhibition of the fungi colonies ranged from 59 to 92%, compared to the untreated control. The B. subtilis isolate SY41B had the highest inhibition effect on the vegetative growth zones as compared with the other species isolates. In view of these, our results indicate that the antagonistic effect of the Bacillus sp. isolates may be important contributors as a biocontrol approach that could be employed as a part of integrated CRR management system.