The ability of microorganisms to attach to living and non-living surfaces and create a biofilm is the cause of numerous long-lasting illnesses, as well as their strong resistance to drugs. Bacterial biofilms consist of intricate assemblies of immobile bacteria. These are located in an extracellular matrix and adhere to various surfaces for a long period. The present study evaluated the antibacterial effectiveness of Plantago major extract against Staphylococcus aureus biofilm. The specimens analyzed in this investigation were skin infections of clinical origin. The current study was not previously studied, particularly in terms of S. aureus biofilm breakdown and inhibition. The disc diffusion method was used to test the antimicrobial activity of extracts on planktonic forms. Several antibiotic control tests were conducted utilizing several commercial disks, The sizes of the inhibitory zones were measured in millimeters and normalized. The anti-biofilm effects were evaluated using the microtiter plate technique. Out of 120 clinical samples, only 46 (38.3 %) had positive S. aureus isolates, whereas 74 (61.6%) were negative. The current study demonstrated that ethanolic P. major leaf extract has antibacterial effects on the development of S. aureus isolates with zones of inhibition ranging in size from 9 to 22 mm. Simultaneously, Antibiotic susceptibility tests showed that all isolates were resistant to erythromycin and clindamycin at an 80 % level. Cefoxitin and chloramphenicol resistance was found at 72 %, and 77 %, respectively. significance and impact of study: this research shown that Extracts from Plantago major can be employed as antibacterial agents against S. aureus, and also anti_staphyloccocus biofilm forms.