Multi-species bacterial biofilm and intracellular infection in otitis media

BMC Pediatrics

Peter C Richmond⁴  Anthony D Keil²  Shyan Vijayasekaran⁵  Harvey L Coates⁵  Jennifer Langlands⁴  Pierre Filion¹  Selma P Wiertsema⁴  Paul J Rigby³  Ruth B Thornton⁴ 

[1] Department of Pathology, PathWest Laboratory Medicine WA, Sir Charles Gairdner Hospital, Western Australia, Australia;Department of Microbiology, PathWest Laboratory Medicine WA, Princess Margaret Hospital for Children, Perth, Western Australia, Australia;Centre for Microscopy, Characterisation and Analysis, The University of Western Australia, Perth, Western Australia, Australia;Vaccine Trials Group, Telethon Institute for Child Health Research, Subiaco, Western Australia, Australia;Department of Otolaryngology, Head and Neck Surgery, The University of Western Australia, Perth, Western Australia, Australia

关键词: transmission electron microscopy;    fluorescent in situ hybridisation;    otitis media;    intracellular infection;    Bacterial biofilm;   

Others  :  1172115
DOI  :  10.1186/1471-2431-11-94

 received in 2011-02-21, accepted in 2011-10-24,  发布年份 2011

【 摘 要 】

Background

Bacteria which are metabolically active yet unable to be cultured and eradicated by antibiotic treatment are present in the middle ear effusion of children with chronic otitis media with effusion (COME) and recurrent acute otitis media (rAOM). These observations are suggestive of biofilm presence or intracellular sequestration of bacteria and may play a role in OM pathogenesis. The aim of this project is to provide evidence for the presence of otopathogenic bacteria intracellularly or within biofilm in the middle ear mucosa of children with COME or rAOM.

Methods

Middle ear mucosal biopsies from 20 children with COME or rAOM were examined for otopathogenic bacteria (either in biofilm or located intracellularly) using transmission electron microscopy (TEM) or species specific fluorescent in situ hybridisation (FISH) and confocal laser scanning microscopy (CLSM). One healthy control biopsy from a child undergoing cochlear implant surgery was also examined.

Results

No bacteria were observed in the healthy control sample. In 2 of the 3 biopsies imaged using TEM, bacteria were observed in mucus containing vacuoles within epithelial cells. Bacterial species within these could not be identified and biofilm was not observed. Using FISH with CLSM, bacteria were seen in 15 of the 17 otitis media mucosal specimens. In this group, 11 (65%) of the 17 middle ear mucosal biopsies showed evidence of bacterial biofilm and 12 demonstrated intracellular bacteria. 52% of biopsies were positive for both biofilm and intracellular bacteria. At least one otopathogen was identified in 13 of the 15 samples where bacteria were present. No differences were observed between biopsies from children with COME and those with rAOM.

Conclusion

Using FISH and CLSM, bacterial biofilm and intracellular infection with known otopathogens are demonstrated on/in the middle ear mucosa of children with COME and/or rAOM. While their role in disease pathogenesis remains to be determined, this previously undescribed infection pattern may help explain the ineffectiveness of current treatment strategies at preventing or resolving COME or rAOM.

【 授权许可】

   
2011 Thornton et al; licensee BioMed Central Ltd.

【 图 表 】

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