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Open Access Highly Accessed Research article

Duodenal and faecal microbiota of celiac children: molecular, phenotype and metabolome characterization

Raffaella Di Cagno1, Maria De Angelis1*, Ilaria De Pasquale1, Maurice Ndagijimana2, Pamela Vernocchi2, Patrizia Ricciuti1, Francesca Gagliardi3, Luca Laghi2, Carmine Crecchio1, Maria Elisabetta Guerzoni2, Marco Gobbetti1 and Ruggiero Francavilla3

Author Affiliations

1 Department of Biologia e Chimica Agro-Forestale ed Ambientale, University of Bari Aldo Moro, via Amendola 165/A, Bari, 70126 Italy

2 Department of Food Science, Alma Mater Studiorum, University of Bologna, via Fanin 46, Bologna, 40127 Italy

3 Department of Pediatrics, University of BariAldo Moro, P.zza Giulio Cesare 11, Bari, 70126 Italy

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BMC Microbiology 2011, 11:219  doi:10.1186/1471-2180-11-219

Published: 4 October 2011

Abstract

Background

Epidemiology of celiac disease (CD) is increasing. CD mainly presents in early childhood with small intestinal villous atrophy and signs of malabsorption. Compared to healthy individuals, CD patients seemed to be characterized by higher numbers of Gram-negative bacteria and lower numbers Gram-positive bacteria.

Results

This study aimed at investigating the microbiota and metabolome of 19 celiac disease children under gluten-free diet (treated celiac disease, T-CD) and 15 non-celiac children (HC). PCR-denaturing gradient gel electrophoresis (DGGE) analyses by universal and group-specific primers were carried out in duodenal biopsies and faecal samples. Based on the number of PCR-DGGE bands, the diversity of Eubacteria was the higher in duodenal biopsies of T-CD than HC children. Bifidobacteria were only found in faecal samples. With a few exceptions, PCR-DGGE profiles of faecal samples for Lactobacillus and Bifidobacteria differed between T-CD and HC. As shown by culture-dependent methods, the levels of Lactobacillus, Enterococcus and Bifidobacteria were confirmed to be significantly higher (P = 0.028; P = 0.019; and P = 0.023, respectively) in fecal samples of HC than in T-CD children. On the contrary, cell counts (CFU/ml) of presumptive Bacteroides, Staphylococcus, Salmonella, Shighella and Klebsiella were significantly higher (P = 0.014) in T-CD compared to HC children. Enterococcus faecium and Lactobacillus plantarum were the species most diffusely identified. This latter species was also found in all duodenal biopsies of T-CD and HC children. Other bacterial species were identified only in T-CD or HC faecal samples. As shown by Randomly Amplified Polymorphic DNA-PCR analysis, the percentage of strains identified as lactobacilli significantly (P = 0.011) differed between T-CD (ca. 26.5%) and HC (ca. 34.6%) groups. The metabolome of T-CD and HC children was studied using faecal and urine samples which were analyzed by gas-chromatography mass spectrometry-solid-phase microextraction and 1H-Nuclear Magnetic Resonance. As shown by Canonical Discriminant Analysis of Principal Coordinates, the levels of volatile organic compounds and free amino acids in faecal and/or urine samples were markedly affected by CD.

Conclusion

As shown by the parallel microbiology and metabolome approach, the gluten-free diet lasting at least two years did not completely restore the microbiota and, consequently, the metabolome of CD children. Some molecules (e.g., ethyl-acetate and octyl-acetate, some short chain fatty acids and free amino acids, and glutamine) seems to be metabolic signatures of CD.