Early Effect of Supplemented Infant Formulae on Intestinal Biomarkers and Microbiota: A Randomized Clinical Trial

Mireille Castanet, Christos Costalos, Nadja Haiden, Jean-Michel Hascoet, Bernard Berger, Norbert Sprenger, Dominik Grathwohl, Harald Brüssow, Nanda De Groot, Philippe Steenhout, Sophie Pecquet, Jalil Benyacoub, Jean-Charles Picaud, Mireille Castanet, Christos Costalos, Nadja Haiden, Jean-Michel Hascoet, Bernard Berger, Norbert Sprenger, Dominik Grathwohl, Harald Brüssow, Nanda De Groot, Philippe Steenhout, Sophie Pecquet, Jalil Benyacoub, Jean-Charles Picaud

Abstract

Background: Post-natal gut maturation in infants interrelates maturation of the morphology, digestive, and immunological functions and gut microbiota development. Here, we explored both microbiota development and markers of gut barrier and maturation in healthy term infants during their early life to assess the interconnection of gut functions during different infant formulae regimes.

Methods: A total of 203 infants were enrolled in this randomized double-blind controlled trial including a breastfed reference group. Infants were fed starter formulae for the first four weeks of life, supplemented with different combination of nutrients (lactoferrin, probiotics (Bifidobacterium animal subsp. Lactis) and prebiotics (Bovine Milk-derived Oligosaccharides-BMOS)) and subsequently fed the control formula up to eight weeks of life. Stool microbiota profiles and biomarkers of early gut maturation, calprotectin (primary outcome), elastase, α-1 antitrypsin (AAT) and neopterin were measured in feces at one, two, four, and eight weeks.

Results: Infants fed formula containing BMOS had lower mean calprotectin levels over the first two to four weeks compared to the other formula groups. Elastase and AAT levels were closer to levels observed in breastfed infants. No differences were observed for neopterin. Global differences between the bacterial communities of all groups were assessed by constrained multivariate analysis with hypothesis testing. The canonical correspondence analysis (CCA) at genus level showed overlap between microbiota profiles at one and four weeks of age in the BMOS supplemented formula group with the breastfed reference, dominated by bifidobacteria. Microbiota profiles of all groups at four weeks were significantly associated with the calprotectin levels at 4 (CCA, p = 0.018) and eight weeks of age (CCA, p = 0.026).

Conclusion: A meaningful correlation was observed between changes in microbiota composition and gut maturation marker calprotectin. The supplementation with BMOS seems to favor gut maturation closer to that of breastfed infants.

Keywords: early gut maturation; infant microbiota; prebiotics; probiotics.

Conflict of interest statement

D.G., N.S., B.B., H.B., N.D.G., S.P. are or were employees of Société des Produits Nestlé S.A., Switzerland. P.S., J.B. are or were employees of Nestlé Health Science. NH reports grants from Medical University of Vienna-Department of Pediatrics, during the conduct of the study. J.M.H., C.C. report grants from Société des Produits Nestlé S.A., outside the submitted work; J.C.P., M.C., have nothing to disclose. The funders declare that they were involved in the design of the study; in the collection, analyses and interpretation of data; in the writing of the manuscript, and in the decision to publish the results.

Figures

Figure 1
Figure 1
Flowchart of infants participating in the study (Intention To Treat, ITT) and 16S (Per Protocol, PP) analysis. * 203 infants were enrolled; one was excluded due to a randomization error. The total number of infants enrolled was 202. GI is gastrointestinal symptoms, w2 and w4 means week 2 and week 4.
Figure 2
Figure 2
Fecal calprotectin z-scores compared to breastfed group set as zero line (PP population).
Figure 3
Figure 3
Gut microbiota composition in infants from the four nutritional intervention groups (PP data set). Determination by 16S rRNA gene sequence analysis: bubble plot of bacteria determined at species level (identified at the right ordinate) for the intervention groups and time points specified on the top abscissa. The proportion of the specified bacterium in the specified sample is indicated by the size of the square (key at top right of the figure).
Figure 4
Figure 4
Canonical correspondence analysis at genus level for the nutritional intervention groups (PP data set). Ellipses indicates the 95% confidence intervals around the centroids. BF, breast fed; F, control formula; FLP, formula lactoferrin probiotic; FLPP, formula lactoferrin probiotic prebiotic.
Figure 5
Figure 5
Anthropometric data. z-scores for body weight, length, head circumference, and BMI (PP data set).

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