Staphylococcus aureus appears a considerable load heavily on the healthcare system due to their many virulence factors, such as, enzymes, toxins, aggregation substances, adhesive surface proteins, and most importantly their ability to form biofilms. These enable the bacterial attachment to host tissue and medical implants, and the establishment of a fully developed physical biofilm which plays a great significant role in the stability of chronic infections. Moreover, biofilms are considered accountable for a wide group of chronic diseases and it was caused by the development of antibiotic resistance in bacteria, it has begun to be a tough task to therapy them with effectiveness. Therefore, new factors are existence searched to keep the action of forming and dissemination of biofilm. Nasal transport of S. aureus is multifactorial and can make someone liable transports to following chronic infections and a danger agent for subsequent infections in nonsurgical patients. This study reports that allfour Eucalyptus, Cinnamon, Clove, and Tea tree essential oils were both antibacterial and antibiofilm against 10 nasal Staphylococcus aureus isolates; Using tube method, eucalyptus and tea tree oils demonstrated high antibiofilm activity as compared with cinnamon and clove. However clove and cinnamon not only exhibited weak biofilm inhibition but at higher concentrations they seemed to enhance it. Microtiter plate method was more sensitive in detecting antibiofilm forming ability of essential oils as it demonstrated that all EOs have antibiofilm activity against nasal Staphylococcus aureus. Our results showed that the TCP method is the most a careful, reliable, and quantitative method for examining biofilm formation by nasal isolates of staphylococci.