The objective of the present work was to study the species composition of microflora in the suppurative exudate from the tympanic cavity in the course of development of experimental suppurative staphylococcal otitis and to identify the initial sites of migration of secondary pathogens. The experiments were carried out on 20 adult rabbits showing no signs of "spontaneous" otitis. Experimental staphylococcal suppurative otitis was induced in 17 of these animals. The microbiological study included isolation and identification of pure bacterial cultures with the use of the classical method. The initial sites of migration of secondary pathogens were detected from the results of comparison of the species composition of microflora in tympanic exudate and the mucous membrane of the nearest anatomical regions, such as the nasal cavity an external auditory canal. The data obtained indicate that suppurative exudate from the tympanic cavity is populated by polyflora containing secondary pathogens, besides the principal ones (Staphylococci). The large amounts of secondary pathogens penetrate into the tympanic cavity from the mucous membranes of the nasal cavity and nasopharynx. It is concluded that the rhinotubal system is the major pathway through which pathogenic microflora migrates into the middle ear.