Salmonella enterica serovar Enteritidis is one of the major food borne pathogens. Utilizing lytic bacteriophages against this pathogen can be a new and effective approach for the prevention of food-contamination and food-borne infection. In this study, we isolated and characterized a Salmonella Enteritidis specific lytic bacteriophage (phage SE2). The bacteriolytic activity of planktonic and biofilmed cells against an antibiotic resistant strain of Salmonella Enteritidis was also evaluated. Phage SE2 revealed an efficient bacteriolytic effect with biofilm dispersing ability and could maintain its virulence even at extreme pH and temperature. It can be a potential biotherapeutic agent against Salmonella Enteritidis.