The current study aimed to investigate the presence of Escherichia coli and Staphylococcus aureus in captive falcons and detection of different serotypes of the isolated Escherichia coli. In addition to performing antibiotic sensitivity test to the isolated bacteria. Fifty diseased falcons of different species (23 Peregrine falcon, 14 Saker falcon and 13 Gyrfalcon) admitted to specialist hospital and research institute of Fahad bin Sultan falcon center, Riyadh, kingdom of Saudi Arabia and private veterinary clinics in Egypt. Thirty-four fecal samples and 28 oropharyngeal samples were examined bacteriologically for the presence of Escherichia coli and Staphylococcus aureus, followed by biochemical and antimicrobial sensitivity test. Moreover, the obtained isolates of Escherichia coli were subjected to serological tests. Overall, Escherichia coli was isolated as (11.77%) and (21.43%) from fecal and oropharyngeal samples respectively while Staphylococcus aureus was isolated as (5.88%) and (7.14%) from the same samples. Mixed infection of both Escherichia coli and Staphylococcus aureus from the fecal and oropharyngeal samples were isolated as (2.94%) and (3.57%) respectively. Six serotypes of Escherichia coli were detected; O55: H7, O103: H2, O17: H18, O126: H21, O78, and O159. Isolates of Escherichia coli showed complete resistance to Cefepime and highly resistance to Cefazolin, Ampicillin, Amoxycillin, Sulphamethoxazol and sensitive to Gentamicin, Meropenem, Amikacin, Imipenem while Staphylococcus aureus showed complete resistance to Cefoxitin, Sulphamethoxazol, Penicillin and highly resistance to Cefotaxime, Amoxycillin, Ampicillin-Sulboctam and highly sensitive to Amikacin.