The present study focused on screening of biosurfactants produced by Leu-conostoc strains isolated from dromedary milk with antimicrobial and antiad-hesive properties against various pathogenic microorganisms and its poten-tial application in medical fields as an antibiofilm agent. After identification of the selected strains by mass spectrometry, screening methods for biosur-factants production were used, including drop collapse, oil displacement, surface tension and emulsification index. Antiadhesive assays were carried out against six pathogenic microorganisms in polystyrene surface, and ap-plied on rubber silicone tubes. Four lactic acid bacteria were isolated and identified as Leuconostoc sp according to MALDITOF MS spectrometry. Bio-surfactants isolated from Leuconostoc strains reduced tension surface from 72.0 to 45.7 by strain BY14, with an emulsification efficiency (E24) after 24h of 63%. The crude biosurfactants were effective against most pathogenic microorganisms with inhibition percentage from 92.3% to 44.3% after adhe-sion to polystyrene. Otherwise, the antiadhesive activity displayed values from 75.67% to 13.2%. The result of application test demonstrated that the selected crude biosurfactants reduced significantly all the deposited microbi-al biofilm from rubber silicone tubes. This study suggest that biosurfactants could have a promising role as an effective antibiofilm to prevent microbial colonization on medical devices.