The histological and microbiological characteristics of bacterial microcolonies in paediatric tonsillar hyperplasia

Chen, Ruyan; Clark, Sita Tarini; Waldvogel-Thurlow, Sharon; Radcliff, Fiona J.; Hoggard, Michael; Johnston, James; Douglas, Richard; Biswas, Kristi

doi:10.1016/j.ijporl.2022.111128

Cited by 2 publications

(1 citation statement)

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“…(PSE227) and Streptococcus spp. (STRC493)) and one species-specific FISH probe ( H. influenzae (HAEinf)) selected based on findings in a previous study by our group 9 . Implementing this technique in tonsil tissue may provide critical insights into differences in the spatial arrangement of potentially pathogenic bacteria within microcolonies for a broad range of aerobic and anaerobic species.…”

Section: Introductionmentioning

confidence: 99%

Fluorescence in situ hybridisation in Carnoy’s fixed tonsil tissue

Clark

Waldvogel-Thurlow

Mackenzie

et al. 2022

Sci Rep

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Fluorescence in situ hybridisation (FISH) is a powerful molecular technique that enables direct visualisation of specific bacterial species. Few studies have established FISH protocols for tonsil tissue in Carnoy’s fixative, accordingly limiting its application to investigate the pathogenesis of tonsillar hyperplasia. Tonsil tissue from 24 children undergoing tonsillectomy for either recurrent tonsillitis or sleep-disordered breathing were obtained during a previous study. The specificity of each of the five FISH probes (Fusobacterium spp., Bacteroides spp., Streptococcus spp., Haemophilus influenzae and Pseudomonas spp.) were successfully optimised using pure and mixed bacterial isolates, and in Carnoy’s fixed tonsil tissue. Bacteroides spp. were present in 100% of patients with microcolonies. In comparison, the prevalence of Fusobacterium spp. was 93.8%, Streptococcus spp. 85.7%, H. influenzae 82.35% and Pseudomonas spp. 76.5%. Notable differences in the organisation of bacterial taxa within a single microcolony were also observed. This is the first study to establish a robust FISH protocol identifying multiple aerobic and anaerobic bacteria in Carnoy’s fixed tonsil tissue. This protocol provides a strong foundation for combining histological and microbiological analyses of Carnoy’s fixed tonsil samples. It may also have important implications on the analysis of microorganisms in other human tissues prepared using the same techniques.

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Section: Introductionmentioning

confidence: 99%