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Background : The food handlers participate in dissemination of typhoid bacilli through different food products and water. People infected by Salmonella species excrete them after the disappearance of symptoms and control of the disease. Such individuals are known as asymptomatic carriers The chronic Salmonella carrier state might be developed either as a sequelae to a typhoid fever or after subclinical infection. The occurrence of subclinical typhoid infection is higher than the acute state, and around 10% of the clearly healthy populations have been observed to be chronic typhoid carriers based on Vi serology in endemic regions Aim : This work was carried out to estimate percentage of S.typhi carriers among food handlers in Mansoura University Hospitals,detect the capsular polysaccharide antigen of S.typhi (Vi antigen) ,a ssess Vi antibody in carriers sera and testing the ability of S. typhi strains isolated from carriers to form biofilm. Setting : Microbiology Diagnostics and Infection Control Unit (MDICU), in the Medical Microbiology and Immunology Department, Faculty of Medicine, Mansoura University. Results : This study was conducted over a period of six months from December 2014 through May 2015 and included 90 asymptomatic food handlers. Stool and urine samples were collected from food handlers with positive Vi Antibody in their sera. Diagnostic Vi antibody was found in the sera of 22 asymptomatic food handlers. S. typhi was isolated from the stool of 15 food handlers carriers (68.18%) and no isolate was recovered from the urine cultures. The carrier state and S. typhi excretion were significantly higher in female than male food handlers working at all tested medical centers at MUHs. As regards the biofilm formation in our study, 40%, 26.66% and 13.33% of S. typhi isolates were strong, moderate and weak biofilm producers, respectively. Our data showed that 75 % of biofilm producing S. typhi were MDR especially in females. Conclusion : The most effective drugs against the isolated S. typhi were cefotaxime and imipenem whereas the least effective was ampicillin and ciprofloxacin. Additionally, the resistance to the first line anti Salmonella drugs was reported in 60% of the isolates.