# In Vivo Study of Probiotic and Anti-Candida Albicans Properties of Lactobacillus Rhamnosus Lcr35®

Cyril Poupet, Adrien Nivoliez, Olivier Camares, Stéphanie Bornes

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Cyril Poupet, Adrien Nivoliez, Olivier Camares, Stéphanie Bornes. In Vivo Study of Probiotic and Anti-Candida Albicans Properties of Lactobacillus Rhamnosus Lcr35®. C. elegans SPAMS Meeting, Jun 2018, Madison, United States. . [hal-01929449](https://hal HAL Id: [hal-01929449](https://hal HAL Id: ) )

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Submitted on 15 Nov 2021

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# In vivo study of probiotic and anti-Candida albicans properties
of Lactobacillus rhamnosus Lcr35®

Cyril POUPET¹, Adrien NIVOLIEZ², Olivier CAMARES¹ and Stéphanie BORNES¹

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1- Université Clermont Auvergne, INRA, UMRF, F-15000, Aurillac, France

2- BIOSE, 24 avenue Georges Pompidou, Aurillac, France

Contact: cyril.poupet@uca.fr

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## Scientific background

Proven probiotic effects of Lcr35®: anti-Candida albicans properties (opportunistic pathogenic yeast in humans) demonstrated by in vitro approaches [1] and clinical trials

→ molecular mechanisms not elucidated

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Caenorhabditis elegans, innovative animal model for studying microorganisms-microorganisms and microorganisms-host interactions

## Methods

### 1- Physiological approaches

Prophylactic treatment:
Lcr35® 2 hours

Infection: C. albicans 2 hours

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Curative treatment:

Infection: C. albicans 2 hours

Lcr35® 2 hours

Worm survival assay

Kaplan-Meier statistical model (log-rank test)

### 2- Targeted transcriptomic approach

Quantification of gene expression by RT-qPCR at different times (2, 4, 6, 24 hours):

- lifespan: *daf-2* (insulin pathway)

- innate immunity: *sek-1* (p38 MAPK pathway)

- antimicrobial: *fipr-22/23*

$$E_{GOI} = \frac{(GOI \text{ efficiency})^{\Delta Ct_{GOI}}}{\sqrt{(cdc-42 \text{ efficiency})^{\Delta Ct_{cdc-42}} \times (Y45F10D.4 \text{ efficiency})^{\Delta Ct_{Y45F10D.4}}}}$$

## Results

### C. elegans lifespan assay

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### Experimental candidiasis in C. elegans : survival assay with a prophylactic ... ... or a curative treatment

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C. albicans : pathogenic (mean lifespan reduced to 7 d)
Lcr35® : beneficial strain (mean lifespan increased to 10 d)

Candida albicans: mean lifespan of 3 d
Lcr35® + C. albicans: mean lifespan of 11 d (+ 267 %)
Lcr35® has a preventive effect against C. albicans

Candida albicans: mean lifespan 3 d
C. albicans + Lcr35®: mean lifespan of 13 d (+ 333 %)
Lcr35® has a curative effect against C. albicans

### daf-2 (insulin pathway)

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OP50: underexpression
Lcr35® (preventive): under- then late overexpression (t = 4h)
Lcr35® (curative): few effects or underexpression

### sek-1 (p38 MAPK pathway)

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OP50: overexpression (↗ in preventive; ↓ in curative)
Lcr35® (preventive): overexpression
Lcr35® (curative): early overexpression (t = 2h) then no effects

### fipr-22 / fipr-23 (antimicrobial)

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OP50: late overexpression (t = 24h)
Lcr35® (preventive): early and stable overexpression
Lcr35® (curative): early (t = 2h) but temporary overexpression

## Conclusion

* Significant protective effects of Lcr35® on *Caenorhabditis elegans* :

* ✓ Inhibition by Lcr35® of *Candida albicans* effects on *C. elegans* survival

* ✓ Effective preventive and curative treatments

* Preliminary results compared to *C. albicans* ATCC 10231:

* ✓ In preventive, overexpression of genes involved in immunity (*sek-1*, *fipr-22/23*), underexpression of the gene involved in longevity (*daf-2*) (except at t = 24 h) in the presence of Lcr35®

* ✓ In curative, similar expression of GOIs in the presence of Lcr35®

## References

[1] Nivoliez A et al. (2012) Influence of manufacturing processes on *in vitro* properties of the probiotic strain *Lactobacillus rhamnosus* Lcr35®. J Biotechnol 160:236-241.

[2] de Barros PP et al. (2018) *Lactobacillus paracasei* 28.4 reduces *in vitro* hyphae formation of *Candida albicans* and prevents the filamentation in an experimental model of *Caenorhabditis elegans*. Microb Pathog 117:80-87.

## Perspectives

* Monitoring the colonization of the worm intestine by the probiotic and the pathogen

* Identification of *C. elegans* signaling pathways modulated by Lcr35® by RNA sequencing followed by a metabolomic study

## Acknowledgments

The doctoral thesis of Cyril Poupet is co-financed by the European Union under the European Regional Development Fund (ERDF), the Auvergne-Rhône-Alpes Region and by biose®.

We thank Jonathan HEUZE, trainee at the UMRF (bioengineering Technological University Degree), for his involvement in carrying out this research.

We thank Taous SARAOUI and Philippe VEISSEIRE for their help during experiments.

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L'EUROPE S'ENGAGE
en région
Auvergne-Rhône-Alpes
avec le FEDER

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UNION EUROPÉENNE

La Région
Auvergne-Rhône-Alpes

biose®
HEALTH THROUGH MICROBIOTA