# Addition of dairy lipids and probiotic Lactobacillus fermentum CECT 5716 in infant formula programs gut microbiota, epithelial permeability, immunity and GLP-1 secretion in adult minipigs

Marion Lemaire, Gaëlle Boudry, Stéphanie Ferret-Bernard, Isabelle Nogret, Michele Formal, Armelle Cahu, Laurence Le Normand, Gwenaëlle Randuineau, Sylvie Guerin, Véronique Rome, et al.

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## ► To cite this version:

Marion Lemaire, Gaëlle Boudry, Stéphanie Ferret-Bernard, Isabelle Nogret, Michele Formal, et al. Addition of dairy lipids and probiotic Lactobacillus fermentum CECT 5716 in infant formula programs gut microbiota, epithelial permeability, immunity and GLP-1 secretion in adult minipigs. 50. Annual Meeting of the European Society for Pediatric Gastroenterology, Hepatology and Nutrition (ESPGHAN), May 2017, Prague, Czech Republic. Journal of Pediatric Gastroenterology and Nutrition, 64 (Suppl. 1), 2017, Journal of Pediatric Gastroenterology and Nutrition. hal-01595024

HAL Id: hal-01595024

https://hal.science/hal-01595024

Submitted on 26 Sep 2017

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# Addition of dairy lipids and probiotic *Lactobacillus fermentum* CECT 5716 in infant formula programs gut microbiota, epithelial permeability, immunity and GLP-1 secretion in adult minipigs

M. Lemaire¹⁻², S. Dou³, G. Boudry¹, S. Ferret-Bernard¹, I. Nogret¹, M. Formal¹, A. Cahu¹, L. Le Normand¹, G. Randuineau¹, S. Guérin¹, V. Romé¹, M. Rhimi⁴, P. Le Ruyet², I. Cuinet², C. Baudry², P. Gérard⁴, S. Blat¹, I. Le Huërou-Luron¹

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INRA
SCIENCE & IMPACT

¹ INRA, INSERM, Univ Rennes 1, Univ Bretagne Loire, Nutrition Metabolisms and Cancer (NuMeCan), Rennes, France ; ² Lactalis R&D, 35240 Retiers, France

³ INRA, UMR1348 PEGASE, Saint-Gilles, France ; ⁴ Micalis Institute, INRA, AgroParisTech, Univ Paris-Saclay, Jouy-en-Josas, France

marion.lemaire@inra.fr

## Context and objective

Postnatal nutrition may have long-lasting metabolic and physiologic impacts in adulthood. Since gut microbiota has been identified as a key factor of this nutritional imprinting, its modulation through infant formula (IF) composition could represent a good strategy to improve the health of formula-fed infants. The addition of dairy lipids (DL) or of a probiotic strain (*Lactobacillus fermentum* CECT 5716 (Lf)) have been associated with benefits in childhood, especially on gut microbiota composition. However, the interaction between DL and Lf on the short- and long-term remains unknown. The objective of this study was therefore to investigate, in a Yucatan minipig model, the long-term effects of the addition of DL and Lf in IF on adult gut microbiota and physiology.

## Methods

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Piglets received from postnatal day (PND) 2 to 28 a formula containing as lipids:

- only plant lipids (PL)

- a half-half mixture of PL and DL (DL)

- a half-half mixture of PL and DL supplemented with Lf (DL+Lf)

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Pigs were subsequently fed:

- a standard diet for 1 month

- then challenged with a hyperenergetic diet (HE) for 3 months

- euthanized at PND140

<figure><img src="image_7.png" /></figure>

### Analyses at PND28 and PND140:

- Gut microbiota composition (16S RNA sequencing)

- Gut microbiota metabolism (¹H NMR)

### Analyses at PND140:

- Intestinal permeability (Ussing chambers)

- Mucosal immunity (cytokine secretion of ileal explants challenged with LPS)

- Endocrine function (density of GLP-1 secreting cells, meal test)

- Metabolism (lipid profile, glucose tolerance (IVGTT))

### Statistics:

- Phenotypic variables: ANOVA testing diet, gender and replication factors followed by post-hoc tests. \* : p < 0.05 and \# : p < 0.1

- Microbiota composition: Edge R

## Results

### 1. Gut microbiota composition and metabolism
In piglets (PND28)

<figure><img src="image_8.png" /><figcaption>**Rectal metabolome**
26 differenciating metabolites</figcaption></figure>

<figure><img src="image_9.png" /></figure>

**In young adults (PND140)**

<figure><img src="image_10.png" /><figcaption>**Rectal metabolome**
5 differenciating metabolites</figcaption></figure>

<figure><img src="image_11.png" /></figure>

The IF composition modulated gut microbiota composition and metabolism on the short- and long-terms, implicating the same main phyla and families at both stages. The effects of DL alone or with Lf were different, the addition of Lf inducing a modulation of more families in the long-term.

### 2. Intestinal permeability (PND140)

<figure><img src="image_12.png" /><figcaption>**Paracellular permeability**
**Ileum**</figcaption></figure>

<figure><img src="image_13.png" /><figcaption>**LPS passage**
**Jejunum**</figcaption></figure>

<figure><img src="image_14.png" /></figure>

The addition of DL+Lf increased intestinal trans- and paracellular permeabilities and prevented LPS passage in the upper gut of young adult minipigs.

### 3. *In vitro* secretion of LPS-stimulated ileal explants (PND140)

<figure><img src="image_15.png" /></figure>

The addition of DL (± Lf) had a beneficial effect on the mucosal immunity of young adults as it decreased pro-inflammatory cytokine secretions.

### 4. Entero-insular axis (PND140)

<figure><img src="image_16.png" /><figcaption>**GLP-1-secreting cells**
**Caecum**</figcaption></figure>

<figure><img src="image_17.png" /></figure>

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The addition of Lf (+DL) had a beneficial effect on the endocrine function in young adulthood by enhancing GLP-1 basal and meal-stimulated secretory capacities.

### 5. Host metabolism (PND140)

<figure><img src="image_19.png" /><figcaption>**Plasma lipid profile**</figcaption></figure>

<figure><img src="image_20.png" /><figcaption>**HOMA-IR**</figcaption></figure>

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FFA: free fatty acids, HOMA-IR: homeostasis model assessment of insulin resistance

The metabolic adaptations to the HE diet were similar between groups

## Conclusion

This study highlights a long-term programming effect of the infant formula composition. This nutritional imprinting, mainly targeting gut microbiota and physiology (barrier, immune and endocrine functions), is different with the addition of dairy lipids alone or associated with the probiotic Lf. Dairy lipids have mainly an impact on the immune function whereas the probiotic Lf has mainly an impact on the barrier and endocrine functions. These long-term effects could be mediated by long-lasting changes in gut microbiota composition and metabolism.

This work was funded by Lactalis Group. The authors acknowledge all the technical staff (UEPR, NGB team) for their expert assistance and help. The authors wish to thank the Nutrition, Chemical Food Safety and Consumer Behaviour research division of the INRA for financing the metabolomics study and the analytical platform for metabolomics and toxicology (C. Canlet, M. Tremblay-Franco, MetaToul-AXIOM, INRA, UMR1331 Toxalim, Toulouse, France) for their help in interpreting these results.