Staphylococcal infections in poultry are taking increasing significance and both coagulase positive Staphylococci (CoPS) and coagulase negative Staphylococci (CoNS) can cause infections in poultry. This study was performed to investigate incidence of CoPS and CoNS and their characteristics among Japanese quails and broiler chickens in some farms at Qena Governorate Therefore, 80 Japanese quail samples of 3-4 weeks old and 70 broiler chicken samples of 21-25 days old were collected. Bacterial isolates were identified as Staphylococci through phenotypic characteristics and PCR assay target 16s rRNA gene of Staphylococci, Bacteriologically, 11 Staphylococcus isolates were isolated from Japanese quail samples with percentage of (13.75%) and they identified as S. aureus (n=2), OCoPS (n=2) and CoNS (n=7) while 5 CoNS isolates were isolated and identified from broiler chicken samples with percentage of (7.14%). Examination of Staphylococcus isolates for their ability to form biofilm by Microtitre plate and Congo Red Agar tests revealed that all isolates were biofilm producer. Antimicrobial susceptibility testing of Staphylococcus isolates revealed that all the isolates were resistant to more than three antibiotic from different family. On the other hand, screening Staphylococcus isolates by PCR for presence of mecA, blaz and vanA resistance genes revealed that (81.25%), (56.25%) and (31.25%) of the isolates harbor these genes respectively and each MRS harbor one gene of mecA and blaz at least. The present study demonstrates that Japanese quails and chickens harbor multidrug-resistant bacteria that could transmit to human.