Background: Food handlers are the most important factors participating in food-borne diseases. Our study aimed at determining the frequency of bacterial and parasitic infections among food handlers in different restaurants in Akharj province-Saudi Arabia. Subject and methods: Total of 65 stool samples and fingernail swaps were taken from participants between September 2019 and December 2019. Wet mount, Gram stain, culture, and biochemical tests, were performed. Conformiation and antimicrobial sensitivity were performed for some samples (microscan). Result: Most of the participants were Indian nationality (26, 40%). About 21 (32%) fingernail and stool samples showed positive culture for different bacterial species, of which, 10% were harboring Staphylococcus aureus (19%) in nail swab. Escherichia coli 11(17%) was the main bacteria isolated from stool specimens, followed by Citrobacter (12%) and Pseudomonas aeruginosa (3%). There was a statistically significant difference between the isolation rate of microorganisms and the work experience and nail status of participants (p = 0.00, and 0.01, respectively). About 17% had intestinal parasites of E. coli (14%) followed by B. Hominis (5%). No parasite was detected in the nails of our study subjects. All Staphylococcus aureus and coagulase-negative Staphylococcus species isolates were uniformly susceptible to vancomycin, 75% of S aureus isolates were resistant to Penicillin. Conclusion: The study revealed a high rate of parasitic and bacterial infections among food handlers, a strict preventive measure should be implemented for personal hygiene and hygienic handling practices of food among food handlers.