Otitis media microbes: culture, PCR, and confocal laser scanning microscopy
T. De Baere*, J. Hollants*, A. Waeytens**, J. Huyghe***, C. Cuvelier**, R. Verhelst*, P. Deschaght*, M. Vaneechoutte* and I. Dhooge***
*Department of Clinical Chemistry, Microbiology and Immunology, **Department of Anatomo-Pathology, ***Department of Oto-rhino-laryngology, Ghent University Hospital, University of Ghent, Gent, Belgium
Otitis media; bacteria; biofilm; culture; PCR
65 - 72
Otitis media microbes: culture, PCR, and confocal laser scanning microscopy. Objectives: To assess the presence of middle ear pathogens in nasopharynx (NP), middle ear fluid (MEF), and middle ear mucosal swabs (MES) of 14 patients undergoing middle ear surgery. Methodology: Bacteria were assessed by culture and species specific PCR. Biofilm was investigated by confocal laser scanning microscopy (CLSM) of middle ear biopsies (MEBs). Results: Bacteria were absent in CLSM of MEBs in three of the four closed and healthy middle ears. Bacteria occurred in the ear with a foreign body (middle ear prosthesis), which showed localized living and dead bacteria, indicating biofilm. Bacterial growth was present in ten patient ears, but biofilm occurred in only one patient. CLSM indicated biofilm in the middle ear of two patients for whom PCR detected Haemophilus influenzae in the MEF. The three classi- cal pathogens could frequently be found in the nasopharynx, by culture and PCR, but not from the middle ear. Alloiococcus otitidis was detected in the MEF of all five patients with open inflamed ears, though virtually absent from the nasopharynx. Pseudomonas aeruginosa was present in seven. It was the only pathogen found on several occasions in all three locations in one patient. Conclusions: This study confirms the association of H. influenzae with middle ear biofilm, and indicates a potential role of P. aeruginosa in middle ear inflammation and biofilm formation. Biofilm does not seem to cause inflammation. It is unclear whether the predominance of A. otitidis in chronically inflamed open middle ears indicates a pathogenic or contaminant role for this organism.
Vol. 5, 2009, 2nd trimester

Otitis media microbes: culture, PCR, and confocal laser scanning microscopy