Background and aims:Thalassemia patients are predisposed to septicemia due to iron overload. The iron chelators have a potential role in the treatment of septicemia. This study was conducted to investigate the in vitro effect of iron chelators, deferoxamine (DFO), deferiprone (DFP), and deferasirox (DFS) on the growth of effective bacteria (Staphylococcus aureus, Yersinia enterocolitica, and Pseudomonas aeruginosa) and their synergistic effect with each other and antibiotics. Materials and Methods: Five bacterial strains were incubated in the presence of seven chelators in Mueller Hinton agar and the number of colonies was recorded for each sample. Antibiotic discs were placed in the plates. Susceptibility to antibiotics was assessed by disc diffusion method (Kirby-Bauer test) and also the zone inhibition around the disc was measured in mm after incubating the relevant plate at 37°C for a day. Results: The growth of all the studied bacteria increased in the presence of iron and was inhibited in the presence of different chelators. Chelators and antibiotics had synergistic effects on growth inhibition of Escherichia coli, S. aureus, and P. aeruginosa in culture medium, while the growth inhibition of Y. enterocolitica was seen in the presence of deferiprone and deferasirox combination. Conclusion: Chelators in combination with antibiotics exert synergistic effects on growth inhibition of prevalent bacteria (except for Y. enterocolitica).