Gluten-degrading microorganisms in adolescents with coeliac disease
Kukovičič, Andraž ...
Oral and gut microbiota play an important role in the pathogenesis of coeliac disease. The aim of our study was isolation and comparison of gluten-degrading microorganisms (GDM) from feces and saliva ...of adolescent patients with coeliac disease (CD) and healthy controls (HC). Additionally, we compared genomes of the same bacterial species isolated from samples of feces and saliva obtained from the same individual. Feces and saliva were obtained from 5 CD patients (2 female, 3 male) on gluten-free diet (GFD) and 5 HC (3 female, 2 male) aged 13-18 years. Samples were inoculated on culturing medium (MCG3) with gluten as a major source of carbon and nitrogen. All colonies with lysis zone were further isolated in pure culture and identified using MALDI Biotyper (Bruker Daltonics). In 4 samples (3 CD, 1 HC), Whole genome sequencing (WGS) was performed on MiSeq platform (Illumina) on all strains that belonged to the same species and were isolated from fecal sample and from saliva in the same individual. In the CD group 10 GDM strains were isolated (5 were not identified): 2 from feces and 8 from saliva. In contrast to the HC group, where 16 GDM were isolated (1 was not identified): 7 from feces and 10 from saliva, 1 GDM was isolated from both samples (saliva and feces). GDM isolated from CD samples belong to 3 genera of bacteria and 1 yeast (Candida albicans). The latter was also isolated in the HC group along with bacteria from 12 different genera. That indicates higher GDM diversity in HC compared with the CD group. Three bacterial species were isolated from feces and saliva of the same individual: Veillonella parvula, Lactobacillus paracasei, Lactobacillus rhamnosus. WGS showed identical genomes only in L. rhamnosus. That could indicate transmission between oral cavity and gut. We found that cultivable GDM are diverse and more often present in feces and saliva of HC than CD, which could be the effect of GFD the CD patients were on. Genomically identical lactobacilli were detected in saliva and in feces of the same individual.
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