Different Fecal Microbiota in Hirschsprung's Patients With and Without Associated Enterocolitis

Alexis P Arnaud, Ianis Cousin, Françoise Schmitt, Thierry Petit, Benoit Parmentier, Guillaume Levard, Guillaume Podevin, Audrey Guinot, Stéphan DeNapoli, Erik Hervieux, Valérie Flaum, Philine De Vries, Gwénaëlle Randuineau, Sandrine David-Le Gall, Sylvie Buffet-Bataillon, Gaëlle Boudry, Alexis P Arnaud, Ianis Cousin, Françoise Schmitt, Thierry Petit, Benoit Parmentier, Guillaume Levard, Guillaume Podevin, Audrey Guinot, Stéphan DeNapoli, Erik Hervieux, Valérie Flaum, Philine De Vries, Gwénaëlle Randuineau, Sandrine David-Le Gall, Sylvie Buffet-Bataillon, Gaëlle Boudry

Abstract

Background and objectives: Patients with Hirschsprung's disease are at risk of developing Hirschsprung-associated enterocolitis, especially in the first 2 years of life. The pathophysiology of this inflammatory disease remains unclear, and intestinal dysbiosis has been proposed in the last decade. The primary objective of this study was to evaluate in a large cohort if Hirschsprung-associated enterocolitis was associated with alterations of fecal bacterial composition compared with HD without enterocolitis in different age groups.

Methods: We analyzed the fecal microbiota structure of 103 Hirschsprung patients from 3 months to 16 years of age, all of whom had completed definitive surgery for rectosigmoid Hirschsprung. 16S rRNA gene sequencing allowed us to compare the microbiota composition between Hirschsprung's disease patients with (HAEC group) or without enterocolitis (HD group) in different age groups (0-2, 2-6, 6-12, and 12-16 years).

Results: Richness and diversity increased with age group but did not differ between HD and HAEC patients, irrespective of the age group. Relative abundance of Actinobacteria was lower in HAEC than in HD patients under 2 years of age (-66%, P = 0.045). Multivariate analysis by linear models (MaAsLin) considering sex, medications, birth mode, breast-feeding, and the Bristol stool scale, as well as surgery parameters, highlighted Flavonifractor plautii and Eggerthella lenta, as well as Ruminococcus gnavus group, as positively associated with Hirschsprung-associated enterocolitis in the 0-2 years age group.

Conclusion: Hirschsprung-associated enterocolitis was associated with features of intestinal dysbiosis in infants (0-2 years) but not in older patients. This could explain the highest rate of enterocolitis in this age group.

Clinical trial registration: https://ichgcp.net/clinical-trials-registry/NCT02857205, MICROPRUNG, NCT02857205, 02/08/2016.

Keywords: Hirschsprung disease; Hirschsprung's associated enterocolitis (HAEC); children; fecal microbiota; proinflammatory bacteria.

Conflict of interest statement

The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Copyright © 2022 Arnaud, Cousin, Schmitt, Petit, Parmentier, Levard, Podevin, Guinot, DeNapoli, Hervieux, Flaum, De Vries, Randuineau, David-Le Gall, Buffet-Bataillon and Boudry.

Figures

Figure 1
Figure 1
MICROPRUNG cohort characteristics. Flowchart of the cohort (A), Hirschsprung-associated enterocolitis incidence as a function of age (B), age at surgery (C), existence of colostomy before surgery (D), length of the resected segment (E), and occurrence of complications more than 1 month after surgery (F). *P < 0.05 for HAEC incidence.
Figure 2
Figure 2
Hirschsprung-associated enterocolitis incidence and factors known to modulate gut microbiota. Hirschsprung-associated enterocolitis incidence as a function of stool consistency (A), delivery mode (B), breast-feeding (C), treatment for transit (D), antibiotic within the last 6 months (E), probiotic treatment (F) or non-steroidal anti-inflammatory drug (NSAID) (G) in the different age groups. *P < 0.05 for Hirschsprung-associated enterocolitis incidence.
Figure 3
Figure 3
Characteristics of fecal microbiota of the MICROPRUNG cohort. Principal coordinates analysis representation of Jaccard distance between fecal microbiota highlighting age group (A) or Hirschsprung-associated enterocolitis status (B). Number of observed species (C) and Shannon index (D) of the microbiota in function of age group and Hirschsprung-associated enterocolitis status. ****P < 0.0001.
Figure 4
Figure 4
Main phyla relative abundances of fecal microbiota in the MICROPRUNG cohort. Relative abundance of Actinobacteria (A), Proteobacteria (B), Bacteroidetes (C), and Firmicutes (D) in function of age group and Hirschsprung-associated enterocolitis status. *P < 0.05.

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