Genomics approach to the analysis of bacterial communities dynamics in Hirschsprung's disease-associated enterocolitis: A pilot study

C. De Filippo, A. Pini-Prato, G. Mattioli, S. Avanzini, G. Rapuzzi, D. Cavalieri, M. Di Paola, I. Stefanini, I. Ceccherini, D. Mavilio, P. Lionetti, V. Jasonni

Research output: Contribution to journalArticle

Abstract

Introduction The most invalidating and life-threatening complication in Hirschsprung's disease patients (HSCR) is Hirschsprung's disease-associated enterocolitis (HAEC). The mechanisms underlying enterocolitis have not been identified. The limited knowledge of the role of intestinal microflora is in part due to the complexity of the intestinal microbiome and to the limitation of cultivation-based technologies, given that less than 25% of the intestinal bacterial species can be cultured. Materials and methods We used amplified ribosomal DNA restriction analysis (ARDRA) with four different restriction enzymes to study variations of microflora composition of the stools of a selected HSCR patient in different clinical conditions (acute phase vs. remission). Results We assessed a total of 15 stool specimens belonging to the same 3-year-old male patient suffering from HSCR, which were harvested during 4 HAEC episodes and remission phases. Restriction analysis showed that HAEC episodes seem to cluster together at ARDRA analysis, thus suggesting a sort of predisposing bacterial community for HAEC development and the need for a microflora equilibrium to maintain wellness. Conclusions This approach proved to be effective, useful and powerful in assessing microflora dynamics and indicated that the differences in microflora associated with acute HAEC or remission are likely to result from a combination of disease activity and different antibiotic therapies. ARDRA proved to be useful in discriminating disease versus remission. Our findings indicated that HAEC results from a change in the equilibrium between bacterial species or from altered discrimination of harmless from harmful microorganisms, challenging the definition of pathogenic and non-pathogenic species. Based on these results, we propose ARDRA as a rapid inexpensive tool to assess microflora dynamics during HAEC episodes.

Original languageEnglish
Pages (from-to)465-471
Number of pages7
JournalPediatric Surgery International
Volume26
Issue number5
DOIs
Publication statusPublished - May 2010

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Enterocolitis
Hirschsprung Disease
Genomics
Ribosomal DNA
Acute Disease
Anti-Bacterial Agents
Technology
Enzymes

Keywords

  • Diagnosis
  • Enterocolitis
  • Genome
  • Hirschsprung
  • Microbioma

ASJC Scopus subject areas

  • Pediatrics, Perinatology, and Child Health
  • Surgery

Cite this

Genomics approach to the analysis of bacterial communities dynamics in Hirschsprung's disease-associated enterocolitis : A pilot study. / De Filippo, C.; Pini-Prato, A.; Mattioli, G.; Avanzini, S.; Rapuzzi, G.; Cavalieri, D.; Di Paola, M.; Stefanini, I.; Ceccherini, I.; Mavilio, D.; Lionetti, P.; Jasonni, V.

In: Pediatric Surgery International, Vol. 26, No. 5, 05.2010, p. 465-471.

Research output: Contribution to journalArticle

De Filippo, C. ; Pini-Prato, A. ; Mattioli, G. ; Avanzini, S. ; Rapuzzi, G. ; Cavalieri, D. ; Di Paola, M. ; Stefanini, I. ; Ceccherini, I. ; Mavilio, D. ; Lionetti, P. ; Jasonni, V. / Genomics approach to the analysis of bacterial communities dynamics in Hirschsprung's disease-associated enterocolitis : A pilot study. In: Pediatric Surgery International. 2010 ; Vol. 26, No. 5. pp. 465-471.
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AU - De Filippo, C.

AU - Pini-Prato, A.

AU - Mattioli, G.

AU - Avanzini, S.

AU - Rapuzzi, G.

AU - Cavalieri, D.

AU - Di Paola, M.

AU - Stefanini, I.

AU - Ceccherini, I.

AU - Mavilio, D.

AU - Lionetti, P.

AU - Jasonni, V.

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AB - Introduction The most invalidating and life-threatening complication in Hirschsprung's disease patients (HSCR) is Hirschsprung's disease-associated enterocolitis (HAEC). The mechanisms underlying enterocolitis have not been identified. The limited knowledge of the role of intestinal microflora is in part due to the complexity of the intestinal microbiome and to the limitation of cultivation-based technologies, given that less than 25% of the intestinal bacterial species can be cultured. Materials and methods We used amplified ribosomal DNA restriction analysis (ARDRA) with four different restriction enzymes to study variations of microflora composition of the stools of a selected HSCR patient in different clinical conditions (acute phase vs. remission). Results We assessed a total of 15 stool specimens belonging to the same 3-year-old male patient suffering from HSCR, which were harvested during 4 HAEC episodes and remission phases. Restriction analysis showed that HAEC episodes seem to cluster together at ARDRA analysis, thus suggesting a sort of predisposing bacterial community for HAEC development and the need for a microflora equilibrium to maintain wellness. Conclusions This approach proved to be effective, useful and powerful in assessing microflora dynamics and indicated that the differences in microflora associated with acute HAEC or remission are likely to result from a combination of disease activity and different antibiotic therapies. ARDRA proved to be useful in discriminating disease versus remission. Our findings indicated that HAEC results from a change in the equilibrium between bacterial species or from altered discrimination of harmless from harmful microorganisms, challenging the definition of pathogenic and non-pathogenic species. Based on these results, we propose ARDRA as a rapid inexpensive tool to assess microflora dynamics during HAEC episodes.

KW - Diagnosis

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KW - Genome

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KW - Microbioma

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