J Clin Microbiol. 2011 Feb 9. [Epub ahead of print]
An Adenoid Reservoir for Pathogenic Biofilm Bacteria.
Nistico L, Kreft R, Gieseke A, Coticchia JM, Burrows A, Khampang P, Liu Y, Kerschner JE, Post JC, Lonergan S, Sampath R, Hu FZ, Ehrlich GD, Stoodley P, Hall-Stoodley L.
Center for Genomic Sciences, Allegheny-Singer Research Institute, Allegheny General Hospital, Pittsburgh, PA; Max-Planck Institute for Marine Microbiology, Bremen, Germany; Wayne State University, Detroit, MI; Medical College of Wisconsin, Milwaukee, WI; Drexel University College of Medicine, Allegheny Campus, Pittsburgh, PA; Ibis Division, Isis Corporation, Carlsbad CA; University of Southampton, National Centre for Advanced Tribology, School of Engineering Sciences, United Kingdom; University of Southampton, Infection, Inflammation & Immunity Division, Faculty of Medicine, NIHR Respiratory BRU & Wellcome Trust Clinical Research Facility, Southampton, United Kingdom.
Biofilms of pathogenic bacteria are present on the middle ear mucosa of children with chronic otitis media (COM) and may contribute to the persistence of pathogens and the recalcitrance of COM to antibiotic treatment. Controlled studies indicate that adenoidectomy is effective in the treatment of COM, suggesting that the adenoids may act as a reservoir for COM pathogens.
To investigate the bacterial community in the adenoid, samples were obtained from 35 children undergoing adenoidectomy for chronic OM or obstructive sleep apnea (OSA). We used a novel, culture-independent molecular diagnostic methodology followed by confocal microscopy (CM) to investigate the in situ distribution and organization of pathogens in the adenoids to determine if pathogenic bacteria exhibited criteria characteristic of biofilms. The Ibis T5000 Universal Biosensor System was used to interrogate the extent of the microbial diversity within adenoid biopsies. Using a suite of 16 broad-range bacterial primers we demonstrated that adenoids from both diagnostic groups were colonized with polymicrobial biofilms. Haemophilus influenzae was present in more adenoids from the COM group (P = 0.005), but there was no significant difference between the two patient groups for Streptococcus pneumoniae or Staphylococcus aureus.
Fluorescence in situ hybridization (FISH), lectin binding and antibodies specific for host epithelial cells demonstrated that pathogens were aggregated, surrounded by a carbohydrate matrix, and localized on and within the epithelial cell surface, consistent with criteria for bacterial biofilms.