The chemical analysis and antibacterial activity of propolis collected from some parts of Western Algeria were investigated. The ethanolic extracts of propolis (EEP) were evaluated for further investigation. The major constituents in EEP were identified by high-performance liquid chromatography (HPLC) analysis. All EEP samples were active against Gram positive bacteria (Staphylococcus aureus, Bacillus subtilis, Bacillus cereus), but no activity was found against Gram negative bacteria (Pseudomonas aeruginosa, Escherichia coli). The mean diameters of growth inhibition of the EEP ranged between 8.05 and 21.4 mm. The propolis extract obtained from Sidi bel Abbés (SFS-SBA) was more active than other samples as well as showed unique HPLC profile. These results support the idea that propolis can be a promising natural food preservative in food industry and alternative candidate for management of bacterial infections caused by drug-resistant microorganisms.