PROBIOTICS:

HOW COMES THEY WORK ?

Goals of this presentation

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- Overview on the latest knowledge on our microbiotas

- Review of few probiotic’s mechanism of action ( non exhaustive list)

Our knowledge is growing

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2012 HUMAN MICROBIOME PROJECT Structure function and diversity of the healthy

human microbiome; N AT U R E | VO L 4 8 6 | 1 4 J U N E 2 0 1 2

5 floras of our body

3 Enterotypes in our gut

5

6

5 enterotypes in the vagina

Vaginal microbiome of

reproductive-age women

Jacques Ravela,1, Pawel

Gajera, Zaid Abdob, G. Maria

Schneiderc, Sara S. K.

Koeniga, Stacey L. McCullea,

et al. Edited by Jeffrey I.

Gordon, Washington

University School of Medicine,

St. Louis, MO, and approved

May 7, 2010 (received for

review March 14, 2010)

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5 Enterotypes in the vagina

Vaginal microbiome of reproductive-age women

Jacques Ravela,1, Pawel Gajera, Zaid Abdob, G. Maria Schneiderc, Sara S. K. Koeniga, Stacey

L. McCullea, et al. Edited by Jeffrey I. Gordon, Washington University School of Medicine, St.

Louis, MO, and approved May 7, 2010 (received for review March 14, 2010)

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All of us similar… but different

Conclusion: “Human microbiology is a complex world”

Few figures…

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25,000 genes 3000 genes

Few figures…

3.300.000 genes

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10,000 billion cells 1,000 billion cells 1 billion cells

Few figures…

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PROBIOTICS:

HOW COMES THEY WORK ?

Development of a

probiotic product

Selection of the best candidate…

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Screening Screening method Intestinal probiotic Vaginal probiotic

Pathogens inhibition X X

Gastric resistance X

Ph 4 resistance x

Bilious resistance X

Adhesion capacity x x

Adhesion inhibition x x

Lactic acid and H202 production

X

Industrial feasability

Human clinicals

The production process…

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Since 1991 (Elo et al., Lett. Appl. Microbiol.) Differences of adhesion of the strain L. rhamnosus GG depending on its product’s origin

2010 (Grześkowiak et al., British J. Nutr.) Inhibitory capacities on the pathogens cellular adhesion of L. GG are significantly different depending on its origin

the manufacturing process influence the characteristics of the strain

impact on clinical trials

2012 (Sảnchez et al., Trends in Food Science & Technology) The development of a dietary probiotic should integrate the influence of the technical constraints.

Bacterial training

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Unlike other probiotic producers, when manufacturing our probiotic we use a unique process called 3Biotic formula. It allows to administrate in the final product not only the probiotic, but as well the prebiotic and its natural postbiotics, integrated in a synergetic compound. The 3Biotic formula is generating a unique efficacy by creating a “bacterial training” effect through the stimulation of specific genes of the probiotic strain. As a result, probiotics coming from 3Biotic formula have an increased efficacy and confirmed in a recently published study(1) outstanding ability to resist to gastric acid and bile, inhibit pathogens and adhere to intestinal wall for biofilm creation.

Increased resistance to gastric acid than native strain

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1,00E+00

1,00E+01

1,00E+02

1,00E+03

1,00E+04

1,00E+05

1,00E+06

1,00E+07

1,00E+08

0 45 min 90 min

UF

C/

ml

Lcr35® Lcr® sachet Lcr® gélule Lcr® Lénio

** ** **∆ ** ** **∆

Resistance to gastric acid : CFU evolution at Ph3

*(1) In vitro comparison of probiotic properties of Lactobacillus rhamnosus Lcr, influences of manufacturing process. / Nivoliez A. et al. / Journal of biotechnology 2012

Bacterial training

Protexin IBS

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-1,42 -1,74

-4,03

-7,87

-6,44

-7,72

-5,87

-6,94 -7,47 -7,42

-8,24 -8,04 -8,35

-7,95 -8,18 -9

-8

-7

-6

-5

-4

-3

-2

-1

0

EPEC ETEC Shigella flexnerfi

Without Lcr Bacilor (Lcr Restituo) Capsule Lcr35 Bacilor (Lcr Restituo) Packet Florea (Lcr Lenio)

(1) (Source : 149. Nivoliez A et al. Influence of manufacturing processes on in vitro properties of probiotic strain Lactobacillus

rhamnosus Lcr35®. Journal of Biotechnology. 2012(in press).

Measure of logarithmic pathogens’ growth after 24h of co-incubation (means ± standard deviation)

Bacterial training

Protexin IBS

0%

10%

20%

30%

40%

50%

60%

70%

80%

Glucose Glycogen

% o

f su

gar

con

sup

tio

n

Lcr35® Lcr Regenerans®

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Bacteria coming from 3biotic formula has an increased

capacity to use Glycogen to produce lactic acid

Usage of glycogen as a substrate

Bacterial training

( Gynophilus)

1,E+00

1,E+01

1,E+02

1,E+03

1,E+04

1,E+05

1,E+06

1,E+07

1,E+08

1,E+09

T0 T4 T24 T28

CF

U / m

l

Temps (h)

vs Lcr35®

vs Lcr Lenio®

vs Lcr Regenerans®

Candida albicansseul

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Inhibition power on Candida albicans

=> Bacteria coming from 3biotic formula totally inhibit

C.albicans in 24h00, when native bacteria does not

Bacterial training

Protexin IBS

( Gynophilus)

Conclusion

The activity of a probiotic is

Strain dependent

Production process dependent

Lcr35

Lcr Regenerans

(Gynophilus)

The secret of probiotics:

interaction

Mechanisms of Action of Probiotics

With our mucosas

With our natural

flora

With our immune system

With our brain

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THEY

INTERACT

Conclusion

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Conclusion

Candiflor1: Clinical study with 409 women. The usage of Gynophilus during 3 weeks gave a 40% reduction rate of candidiasis recurrence on a 6 months observational period.

Dapoigny et al: intake of Lcr35 in IBS-D patients implicated a 36% reduction of Francis score, with a remanence of at least 15 days…

Mechanism of action of probiotics is an extremely complex subject. The more we know… the less we know.

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Conclusion

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Conclusion