title: Characterization of mucosal biofilms on human adenoid tissues.
abstract: OBJECTIVE
  To demonstrate the presence of mucosal biofilm in adenoid tissue using double staining for visualization of both the bacterial matrix and the bacterial cells. To identify bacterial species present on the surface of the studied adenoids.
  
  
  METHODS
  Prospective study.
  
  
  METHODS
  A total of 39 specimens of adenoidectomy were removed from children with chronic and/or recurrent otitis media. The specimens were prepared for light microscopy using Gram staining, scanning electron microscopy (SEM) and confocal laser scanning microscopy (CLSM). Double staining was performed with CLSM to visualize both the bacteria and the glycocalyx matrix. Nine adenoids on which bacterial biofilms were visualized with CLSM were used for identification of bacterial species by 16S-DNA polymerase chain reaction (PCR) amplification and homology analysis.
  
  
  RESULTS
  Of the 39 adenoids investigated, 22 (54%) showed evidence of mucosal biofilms. Gram staining, SEM and CLSM showed the presence of bacterial cells, organized in bacterial microcolonies. CLSM with double staining demonstrated mucosal biofilms by showing the presence of both bacteria and the glycocalyx. The use of 16S-DNA polymerase chain reaction (PCR) amplification and subsequent sequence analyses identified the presence of Corynebacterium argentoratense, Streptococcus salivarius, Micrococcus luteus, and Staphylococcus aureus.
  
  
  CONCLUSIONS
  This study demonstrates that adenoid tissue in children with chronic or/and recurrent otitis media contains mucosal biofilms in 54% of the cases. The existence of living bacteria has been demonstrated. Further studies are required to describe the panel of bacteria that can be harbored in the biofilms present in adenoids and the mechanisms involved in the physiopathology of otitis prone children.
authors.affiliation: Department of Oto-Rhino-Laryngology, Head & Neck Surgery, Leiden University Medical Center, Leiden, The Netherlands. romain.kania@lrb.aphp.fr
authors.forename: Romain E
authors.initials: RE
authors.lastname: Kania
authors.forename: Gerda E M
authors.initials: GE
authors.lastname: Lamers
authors.forename: Marcel J
authors.initials: MJ
authors.lastname: Vonk
authors.forename: Esmee
authors.initials: E
authors.lastname: Dorpmans
authors.forename: Joyce
authors.initials: J
authors.lastname: Struik
authors.forename: Patrice
authors.initials: P
authors.lastname: Tran Ba Huy
authors.forename: Pieter
authors.initials: P
authors.lastname: Hiemstra
authors.forename: Guido V
authors.initials: GV
authors.lastname: Bloemberg
authors.forename: Jan J
authors.initials: JJ
authors.lastname: Grote
mesh_terms: D000233:Adenoidectomy; D000234:Adenoids / Q000382:microbiology*; D018441:Biofilms / Q000145:classification*; D002648:Child; D002908:Chronic Disease; D004396:Coloring Agents; D003352:Corynebacterium / Q000145:classification / Q000502:physiology; D004269:DNA, Bacterial / Q000032:analysis; D019276:Glycocalyx / Q000382:microbiology / Q000648:ultrastructure; D006801:Humans; D007091:Image Processing, Computer-Assisted; D016982:Micrococcus luteus / Q000502:physiology; D018613:Microscopy, Confocal; D008855:Microscopy, Electron, Scanning; D010033:Otitis Media / Q000382:microbiology / Q000601:surgery; D011446:Prospective Studies; D012008:Recurrence; D013211:Staphylococcus aureus / Q000502:physiology; D013291:Streptococcus / Q000145:classification / Q000502:physiology
chemical_list: D004396:Coloring Agents; D004269:DNA, Bacterial