The neutrophil chemotactic activity of middle ear effusions (MEE) was measured in patients with pediatric chronic, adult chronic, and acute otitis media with effusion (OME). Chemotactic activity differed significantly among the three groups of otitis media in the following order: adult chronic less than pediatric chronic less than acute. Culture-positive effusions had higher chemotactic activity than did those that tested culture negative, suggesting that bacterial infection is related to neutrophil chemotactic activity. The chemotactic function of normal peripheral neutrophils was not enhanced by preincubation with MEE, but their bactericidal function was enhanced. In conclusion, bacterial infection in the middle ear is one of the major determining factors of neutrophil infiltration during OME, and MEE enhances the bactericidal function of neutrophils.