Background : Otitis Media continues to be a major presentation in the ear, nose and throat clinic. Aim : This study aimed to isolate, characterize and identify the bacteriological and mycological etiologic agents of otitis media in Benin city. Patients and Methods : Ear discharge from 569 (299 males and 270 females) patients diagnosed clinically of otitis media between August 2009 and August 2010 were processed to recover the bacterial and fungal etiologic agents. Susceptibility test was performed on all bacterial isolate. Result : Pseudomonas aeruginosa (28.3%) was the predominant bacteria isolate causing otitis media followed by Staphylococcus aureus (21.0%), Klebsiella sp (8.9%), Proteus sp (8.2%), Alkaligenes spp (4.3%), Streptococcus pneumoniae (3.9%), Escherichia coli (3.0%) and Citrobacter freundi (1.7%). Fungi isolated were Aspergillus niger (9.2%), Candida albicans (5.4%), Candida tropicalis (3.0%), Aspergillus flavus (2.1%) and Candida parasilopsis (1.5%). 413 had a single organism isolated from the middle ear culture while twenty (3.51%) patients had mixed organisms isolated. Infection was highest among 0 - 5 years, and lowest among aged 18 - 23. All bacterial isolates were poorly susceptible to the antibacterial agents. Conclusion : The study uncovers a high frequency of bacteria associated otitis media with the finding of fungi too as a significant etiologic agent.