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454 Pyrosequencing Analysis on Faecal Samples from a Randomized DBPC Trial of Colicky Infants Treated with Lactobacillus reuteri DSM 17938

Author

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  • Stefan Roos
  • Johan Dicksved
  • Valentina Tarasco
  • Emanuela Locatelli
  • Fulvio Ricceri
  • Ulf Grandin
  • Francesco Savino

Abstract

Objective: To analyze the global microbial composition, using large-scale DNA sequencing of 16 S rRNA genes, in faecal samples from colicky infants given L. reuteri DSM 17938 or placebo. Methods: Twenty-nine colicky infants (age 10–60 days) were enrolled and randomly assigned to receive either Lactobacillus reuteri (108 cfu) or a placebo once daily for 21 days. Responders were defined as subjects with a decrease of 50% in daily crying time at day 21 compared with the starting point. The microbiota of faecal samples from day 1 and 21 were analyzed using 454 pyrosequencing. The primers: Bakt_341F and Bakt_805R, complemented with 454 adapters and sample specific barcodes were used for PCR amplification of the 16 S rRNA genes. The structure of the data was explored by using permutational multivariate analysis of variance and effects of different variables were visualized with ordination analysis. Results: The infants’ faecal microbiota were composed of Proteobacteria, Firmicutes, Actinobacteria and Bacteroidetes as the four main phyla. The composition of the microbiota in infants with colic had very high inter-individual variability with Firmicutes/Bacteroidetes ratios varying from 4000 to 0.025. On an individual basis, the microbiota was, however, relatively stable over time. Treatment with L. reuteri DSM 17938 did not change the global composition of the microbiota, but when comparing responders with non-responders the group responders had an increased relative abundance of the phyla Bacteroidetes and genus Bacteroides at day 21 compared with day 0. Furthermore, the phyla composition of the infants at day 21 could be divided into three enterotype groups, dominated by Firmicutes, Bacteroidetes, and Actinobacteria, respectively. Conclusion: L. reuteri DSM 17938 did not affect the global composition of the microbiota. However, the increase of Bacteroidetes in the responder infants indicated that a decrease in colicky symptoms was linked to changes of the microbiota. Trial Registration: ClinicalTrials.gov NCT00893711

Suggested Citation

  • Stefan Roos & Johan Dicksved & Valentina Tarasco & Emanuela Locatelli & Fulvio Ricceri & Ulf Grandin & Francesco Savino, 2013. "454 Pyrosequencing Analysis on Faecal Samples from a Randomized DBPC Trial of Colicky Infants Treated with Lactobacillus reuteri DSM 17938," PLOS ONE, Public Library of Science, vol. 8(2), pages 1-5, February.
    • Handle: RePEc:plo:pone00:0056710
    DOI: 10.1371/journal.pone.0056710
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    Cited by:

    1. Åsa Vilson & Ziad Ramadan & Qinghong Li & Åke Hedhammar & Arleigh Reynolds & Julie Spears & Jeff Labuda & Robyn Pelker & Bengt Björkstén & Johan Dicksved & Helene Hansson-Hamlin, 2018. "Disentangling factors that shape the gut microbiota in German Shepherd dogs," PLOS ONE, Public Library of Science, vol. 13(3), pages 1-16, March.
    2. Man Xu & Jiao Wang & Ning Wang & Fei Sun & Lin Wang & Xiao-Hong Liu, 2015. "The Efficacy and Safety of the Probiotic Bacterium Lactobacillus reuteri DSM 17938 for Infantile Colic: A Meta-Analysis of Randomized Controlled Trials," PLOS ONE, Public Library of Science, vol. 10(10), pages 1-16, October.

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