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Updated: Jan 27, 2026

Paraffin Embedding and Thin Sectioning of Microbial Colony Biofilms for Microscopic Analysis
Published on: March 23, 2018
Cross-sectional study of adenoidal biofilms in a paediatric population and its clinical implications
Joao Subtil1, Maria Paula Bajanca-Lavado2, Joao Rodrigues3
1Serviço de otorrinolaringologia (ORL), Hospital de Beatriz Ângelo (HBA), Loures, Portugal.
Insights
Biofilms on adenoids were found in over 27% of samples, but their presence did not correlate with recurrent respiratory infections in children. In vitro biofilm formation by bacteria also did not predict in vivo assembly or antibiotic resistance.
Area of Science:
- Microbiology
- Otolaryngology
- Infectious Diseases
Background:
- Adenoids are lymphoid tissues crucial for upper respiratory tract defense but can harbor microbes causing infections.
- Recurrent infections and otitis are common in children, with adenoids potentially playing a role as a microbial reservoir.
Purpose of the Study:
- To compare biofilm presence on adenoids with recurrent infections in children undergoing adenoidectomy.
- To investigate the association between in vitro biofilm formation, antibiotic susceptibility, and in vivo biofilm presence on adenoid tissues.
Main Methods:
- Scanning electron microscopy for in vivo biofilm assessment.
- Crystal violet assay for in vitro biofilm quantification.
- Antibiotic susceptibility testing and PCR for bacterial typing.
Main Results:
- Biofilms were detected on 27.4% of adenoid samples, irrespective of infectious or non-infectious indications.
- Most common bacteria were moderate biofilm assemblers in vitro, with significant antibiotic intermediate/resistance.
- No correlation was found between in vitro biofilm assembly and in vivo presence, nor with antibiotic resistance.
Conclusions:
- Adenoid biofilm presence is independent of clinical indication.
- In vitro bacterial biofilm formation does not predict in vivo biofilm assembly on adenoids.
- The study questions the direct role of biofilms in the pathogenesis of recurrent respiratory tract infections.
Introduction:
Adenoids are nasopharyngeal lymphoid tissue with a relevant role in host defence against infection of upper respiratory tract. Nevertheless, adenoids are also a reservoir of microorganisms that can cause infections of upper respiratory tract and otitis particularly in children.
Objective:
Evaluate and compare the association between biofilm assembly on adenoids and the incidence of recurrent infections in a paediatric population submitted to adenoidectomy by either infectious or non-infectious indication.
Methods:
Scanning electron microscopy was used to assess biofilms on adenoid surface; biofilm assembly in vitro was monitored by crystal violet assay; antibiotic susceptibility was assessed following EUCAST guidelines; Hinfluenzae capsular typing was performed by PCR.
Results:
Biofilms were present in 27.4% of adenoid samples and no statistical difference was found between infectious and non-infectious groups. In vitro, the most clinically relevant bacteria, H.influenzae, S.aureus, S.pyogenes, S.pneumoniae and M.catarrhalis, were mostly moderate biofilm assemblers (71.7%). 55.3% of these bacteria were intermediate/resistant to at least one of the tested antibiotics. No association was found between the ability to assemble biofilms in vitro and the presence of biofilms on adenoids nor antibiotic resistance. All H.influenzae were characterized as non-typeable.
Conclusion:
The presence of biofilms on adenoid surface was independent from clinical sample background. Bacterial ability to assemble biofilms in vitro cannot be used to predict biofilm assembly in vivo. The lack of correlation between biofilm formation and infectious respiratory diseases found contributes to question the relevance of biofilms on the pathogenesis of infectious diseases.
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