¹Department of Pharmaceutical Microbiology, Faculty of Pharmacy, Ankara University, Yenimahalle 06560 Ankara, Turkey DOI : 10.29228/jrp.682 Salmonella is an important agent that causes foodborne gastrointestinal infections. Salmonella enterica serovar Enteritidis, serovar Typhimurium, and serovar Infantis are the main pathogenic agents of several enteric infections worldwide. Due to the emergence of multidrug resistant Salmonella strains, there is a need for new administration strategies alternative to antibiotics, such as bacteriophages, for effective control of them. The main objectives of this study were to isolate and characterize Salmonella spp. bacteriophage and compare its activity to commercial phage product and assess the effect of bacteriophages in vitro during 24-h. Because of the study, it was found that the newly isolated and named vB_SiM_12 phage could lyse 100% of the strains tested. Bacterial growth with dilutions of vB_SiM_12 phage (MOI ranging from 0.001 to 10) was measured during 24-h. The optic density values of the samples with added bacteriophage remained lower than the control sample during the 24-h incubation period. The results show that using phages may offer a promising alternative to combat biological control agents against Salmonella infection. Keywords : Bacteriophage; bacteriolytic activity, invitro turbidity test; Salmonella spp..