Abstract: Lactobacillus is a genus of lactic acid bacteria described as a heterogeneous group of regular non spore forming gram positive rods and found in a great variety of habitats such as plants and gastrointestinal tracts. The aim of this study was to isolate lactobacilli from plants and determine their inhibitory effect against some pathogens. Sixty lactobacilli isolates from fresh vegetables were enriched in Man-Rogosa-Sharpe medium (MRS) broth and isolated by growing on MRS agar medium and characterized by phenotypic characteristics and PCR technique at genus and species levels. The antimicrobial substance was extracted with ethyl acetate solvent and the antimicrobial activity against some pathogenic bacteria such as Escherichia coli, Salmonella typhi, Shigella dysenteriae, Bacillus anthracis and Staphylococcus aureus were investigated. Forteen L. plantarum and eight L. casei which were isolated from fresh vegetables were identified by PCR. Antimicrobial substance from MRS broth medium was extracted. This antimicrobial compound showed a potent inhibitory activity against all tested bacteria. The inhibitory substance was distinct from bacteriocins, lactic and acetic acids which are produced by these bacteria. In conclusion, fresh vegetables may be used as a source of antimicrobial lactic acid bacteria. L. casei and L. plantarum as two probiotics can establish themselves in gut and urogenital tract and prevent the human body from adverse effects of pathogens.