Background: Staphylococcus spp. are widely distributed in nature and can cause nosocomial, skin infections, and foodborne illness, and it may lead to severe financial losses in birds by causing systemic infection in numerous organs. Aim: This study was conducted to determine the prevalence of Staphylococcus spp. in humans and birds in Baghdad city. Methods: Seventy-six oral cavity swabs were collected, including 41 from birds and 35 from breeders. All samples were examined by bacteriological methods and identified by using the VITEK technique, the samples were then further studied to test the ability of biofilm formation, and MDR factors and MAR index were tested with the use of seven antibiotics. Results: Among the 76 oral swabs, 37 samples were positive (48.68%) for Staphylococcus spp.: 7 human samples (20%) and 30 bird samples (73.17%). In humans, S. lentus was the most prevalent (42.85%) followed by S. aureus (28.57%), S. hominis and S. sciuri were at (14.29%) to each. In birds, S. pseudintermedius, S. gallinarum, S. lentus, S. haemolyticus, Staphylococcus spp, S. sciuri and S. xylosus were detected in 36.67%, 16.67%, 10%, 10%, 13.33%, 3.33% and 3.33% respectively. Staphylococcus isolates from the human samples demonstrated that only S. lentus was resistant 33.33% to ME, OX, and SXT. Furthermore, one of them was MDR and high MAR index value. The antimicrobial pattern of Staphylococcus spp. isolated from birds was as follows: S. pseudintermedius isolates demonstrated 100% resistance to CN, CIP, SXT, and MDR (100%) and high MAR indices value; S. xylosus was resistant 100% against ME, CN, SXT and Do and it was MDR with high MAR index;S. lentus was resistant 25% against ME, OX, C, and SXT, whereas, S. gallinarum was resistant 33.33% against ME and OX. The results demonstrated that biofilm formation of the Staphylococcus spp. isolated from human samples were weak biofilm formers: S. lentus, S. hominis, and S. aureus, while other S. aureus (50%) was moderate. In birds, the majority of the isolates had non biofilm-producing capabilities, while 80% of S. lentus and 100% of S. xylosus showed moderate biofilm formation. Conclusion: Healthcare problem was observed in this study due to high MDR and MAR index among Staphylococcus spp. isolated from pet birds to their owners and vice versa.