Probiotics goes beyond digestive and immune health. At the recent Microbiome R&D and Business Collaboration Congress: Asia, DuPont Nutrition & Health shared on how probiotics can influence vaginal microbiota and provide positive health benefits such as weight management for the user.

1. D u P o n t N u t r i t i o n & H e a l t h
March 6, 2019
Dietary tools for shaping
the gut microbiome
Anders Henriksson, Ph.D.
Principal Application Specialist, ANZ

2. D u P o n t N u t r i t i o n & H e a l t h 2
The gut microbiota is a complex community of microorganisms
which colonizes the gastrointestinal (GI) tract
Lagier et al., 2012
Contain bacteria, archaea,
fungi, protozoa and viruses.
~1000 species
1013-1014 microbes
10-fold more than human cells
>100-fold more genes than the
human host

3. D u P o n t N u t r i t i o n & H e a l t h 3
Influences on microbiome composition
• The gut microbiome is associated with co-
variants
• Host-intrinsic - Host genetics
• Host-extrinsic – Lifestyle
• Environmental - Regional strain pools
• Taxa Interactions -
Competition/Cooperation
• Variations in microbiome composition are not
fully explainable
• Inter-individual microbiome variation is
associated with differential responses to
interventions
Figure from Schmidt, et al. (2018) Cell

4. D u P o n t N u t r i t i o n & H e a l t h
The Difference a Healthy Microbiome Makes!
• The gut microbiome – changes throughout life, focusing on Bifidobacterium spp.
• Members of the microbiome and their potential role in
− Reducing prevalence of gut infections
− Modulating blood lipids
− Body weight
− Metabolic health
− Immune health later in life
• Dietary tools for making a difference
Overview
4

5. D u P o n t N u t r i t i o n & H e a l t hD u P o n t N u t r i t i o n & H e a l t h 5
Dose-response effect of Bifidobacterium lactis HN019 on whole gut transit time
and functional gastrointestinal symptoms in adults.
Waller et al (2011) Scand J Gastroenterol.
Beneficial effects of Bifidobacterium lactis on lipid profile and cytokines in patients with
metabolic syndrome: A randomized trial. Effects of probiotics on metabolic syndrome.
Bernini et al (2016) Nutrition
Supplementation with Bifidobacterium lactis probiotics in
pregnancy increases levels of TGF-ᵝ and IgA in breast milk
Prescott et al (2008) Clinical and Experimental Allergy
Prebiotic and Probiotic Fortified Milk in Prevention of Morbidities among
Children: Community-Based, Randomized, Double-Blind, Controlled Trial
Sazawal et al (2010) PlusOne
Probiotic with or without Fiber Controls Body Fat Mass, Associated With Serum Zonulin, in Overweight
and Obese Adults. Stenman et al (2016) EBioMedicine.
Bifidobacterium lactis reduces on reduces risk for Diarrhea
and Fever in Preschool Children in an Urban Slum in India.
Hemalatha et al (2014) Eur. J. Nutrition & Food Safety
The Effect of Bifidobacterium animalis HN019 on Cellular Immune Function in
Healthy Elderly Subjects: Systematic Review and Meta-Analysis
Miller et al (2017) Nutrients
Proposed beneficial effects of Bifidobacterium animalis lactis strains

6. D u P o n t N u t r i t i o n & H e a l t h 6
• 624 children (1-3 years)
• A double blind randomized, 12 months, India*
• Two arms -
• Active: Milk supplemented with Bifidobacterium lactis HN019 (2E+07 CFU/day) in
reconstituted milk with 2.5 g GOS
• Placebo: Milk
• Outcomes measured - Diarrhoea/dysentry, pneumonia, lower respiratory
infections
Sazawal et al (2010) PloSone. 5:1-8
Placebo
Treatment
12 months
Weekly Health assessment
6 months
Blood sample Blood sample Blood sample
The effect of Bifidobacterium lactis HN019TM, in combination with GOS, on
the prevalence of dysentery, pneumonia and LRTI

7. 7D u P o n t N u t r i t i o n & H e a l t h
Sazawal et al (2010) PloSone. 5:1-8
Impact of probiotic consumption was significant
16%
24%
21%

8. 8D u P o n t N u t r i t i o n & H e a l t h
Effect of B. lactis HN019TM on the incidence of diarrhea and fever
• Double-blind, randomized, controlled
trial
• 245 healthy preschool children in an
urban slum in Hyderabad, India
• Daily dose 5x109 CFU/day or placebo
for nine months
Hemalatha et al. Eur. J. Nutr. Food Safety

9. D u P o n t N u t r i t i o n & H e a l t h 9
Effect of Bifidobacterium lactis HN019TM on the prevalence of diarrhea
and fever in India
August – September
*Significantly different from placebo (p < 0.05)
*
*

10. D u P o n t N u t r i t i o n & H e a l t h 10
ESTABLISHING A HEALTHY MICROBIOME IN EARLY LIFEHuman milk oligosaccharides
• A group of unique oligosaccharides found in
human milk.
• HMOs are the 3rd –LARGEST SOLUTE IN
HUMAN MILK after lactose and fat.

11. D u P o n t N u t r i t i o n & H e a l t h 11
HMOs HELP SHAPE A HEALTHY MICROBIOTA
Prebiotic effects – Influencing the human microbiome
HMOs ARE IMPORTANT BIOACTIVES that promote and maintain
health from birth through early life
• HMOs are highly bifidogenic and promote
a group of bifidobacteria that typically
colonize infants.
• Bifidobacterium infantis
• Bifidobacterium longum
Underwood et al, 2014

12. D u P o n t N u t r i t i o n & H e a l t h 12
HMOs HELP SHAPE A HEALTHY MICROBIOTA
Cognitive health
Inhibition of pathogens adhering to
enterocytes
Viral anti-adhesion
Immune-modulating effects
Respiratory health effects
Beneficial metabolite production
Prebiotic effects – Influencing the human microbiome
HMOs ARE IMPORTANT BIOACTIVES that promote and maintain
health from birth through early life
• HMOs are highly bifidogenic and promote
a group of bifidobacteria that typically
colonize infants.
• Bifidobacterium infantis
• Bifidobacterium longum
• HMOs strengthen the gut barrier function
and reduce ability of pathogens to adhere to
the mucosa
• HMOs stimulate the immune system.

13. D u P o n t N u t r i t i o n & H e a l t hD u P o n t N u t r i t i o n & H e a l t h 13
Dose-response effect of Bifidobacterium lactis HN019 on whole gut transit time
and functional gastrointestinal symptoms in adults.
Waller et al (2011) Scand J Gastroenterol.
Beneficial effects of Bifidobacterium lactis on lipid profile and cytokines in patients with
metabolic syndrome: A randomized trial. Effects of probiotics on metabolic syndrome.
Bernini et al (2016) Nutrition
Supplementation with Bifidobacterium lactis probiotics in
pregnancy increases levels of TGF-ᵝ and IgA in breast milk
Prescott et al (2008) Clinical and Experimental Allergy
Prebiotic and Probiotic Fortified Milk in Prevention of Morbidities among
Children: Community-Based, Randomized, Double-Blind, Controlled Trial
Sazawal et al (2010) PlusOne
Probiotic with or without Fiber Controls Body Fat Mass, Associated With Serum Zonulin, in Overweight
and Obese Adults. Stenman et al (2016) EBioMedicine.
Bifidobacterium lactis reduces on reduces risk for Diarrhea
and Fever in Preschool Children in an Urban Slum in India.
Hemalatha et al (2014) Eur. J. Nutrition & Food Safety
The Effect of Bifidobacterium animalis HN019 on Cellular Immune Function in
Healthy Elderly Subjects: Systematic Review and Meta-Analysis
Miller et al (2017) Nutrients
Proposed beneficial effects of Bifidobacterium animalis lactis strains

14. D u P o n t N u t r i t i o n & H e a l t h 14

15. D u P o n t N u t r i t i o n & H e a l t h 15
SPI 1 = SUPRO® 670
SPI 2 = SUPRO® 760

16. 16D u P o n t N u t r i t i o n & H e a l t h
Effects dietary proteins on blood and serum cardio metabolic measures
Butteiger (2016) J Nutr. 146:697-705

17. D u P o n t N u t r i t i o n & H e a l t h 17
Spearman correlations of microbial families and lipid parameters
Butteiger (2016) J Nutr. 146:697-705

18. D u P o n t N u t r i t i o n & H e a l t h 18
Cholesterol-lowering effect of soy protein is well established
(45%) (80%) (66%) (89%) (60%) (61%) (59%)
Anderson et al.
1995. NEJM
333:276;
Harland et al. 2012.
in Soy: Nutrition,
Conump. & Htl.
Reynolds et al.,
2006. Am J
Card 98:633;
Zhan & Ho.
2005. AJCN
81:397.
Balk et al.
2005. AHRQ
No 05-E024-2;
Harland & Haffner.
2008. Atherosclerosis
200: 13;
Anderson et al.
2011 J Am Coll
Nutr 30: 79-91;
• Soy’s ability to lower plasma cholesterol has been widely studied and well established
• Multiple meta-analyses have been published confirming the cholesterol lowering effects in humans
13 countries have an
approved health claim
for soy protein
Japan
United States
Canada
India
South Africa
Brazil
Chile
Colombia
Philippines
Indonesia
Malaysia
Korea
Turkey

19. D u P o n t N u t r i t i o n & H e a l t hD u P o n t N u t r i t i o n & H e a l t h 19
Dose-response effect of Bifidobacterium lactis HN019 on whole gut transit time
and functional gastrointestinal symptoms in adults.
Waller et al (2011) Scand J Gastroenterol.
Beneficial effects of Bifidobacterium lactis on lipid profile and cytokines in patients with
metabolic syndrome: A randomized trial. Effects of probiotics on metabolic syndrome.
Bernini et al (2016) Nutrition
Supplementation with Bifidobacterium lactis probiotics in
pregnancy increases levels of TGF-ᵝ and IgA in breast milk
Prescott et al (2008) Clinical and Experimental Allergy
Prebiotic and Probiotic Fortified Milk in Prevention of Morbidities among
Children: Community-Based, Randomized, Double-Blind, Controlled Trial
Sazawal et al (2010) PlusOne
Probiotic with or without Fiber Controls Body Fat Mass, Associated With Serum Zonulin, in Overweight
and Obese Adults. Stenman et al (2016) EBioMedicine.
Bifidobacterium lactis reduces on reduces risk for Diarrhea
and Fever in Preschool Children in an Urban Slum in India.
Hemalatha et al (2014) Eur. J. Nutrition & Food Safety
The Effect of Bifidobacterium animalis HN019 on Cellular Immune Function in
Healthy Elderly Subjects: Systematic Review and Meta-Analysis
Miller et al (2017) Nutrients
Examples of reported benefits of Bifidobacterium animalis lactis

20. D u P o n t N u t r i t i o n & H e a l t h 20
Akkermansia muciniphila and metabolic health
Inflammation and onset of type 2-
diabetes, metabolic syndrome
LPS

21. 21D u P o n t N u t r i t i o n & H e a l t h
High levels of Akkermansia muciniphila supports good metabolic health
Adapted from Dao et al (2016) Gut 65:426-436

22. D u P o n t N u t r i t i o n & H e a l t h 22
Certain probiotic strains improves mucosal integrity
Putaala et al. 2008
DM Ctrl B. lactis 420™ B. lactis HN019™
The effect of cell free culture supernatants on the
trans-epithelial electrical resistance (TEER).
*Statistically different from the control (p<0.05)

23. D u P o n t N u t r i t i o n & H e a l t h 23
Stenman et al. Beneficial Microbes 2014
The effect of B. lactis B-420™ on plasma LPS levels
0
1
2
3
4
5
6
7
NC Control B-420™
a
b
a
HFD
PlasmaLPS
(EU/ml)
Gut barrier function in
diabetic mice fed with a
high fat diet (HFD) and
treated with the potential
probiotic Bifidobacterium
animalis ssp. lactis 420
(B-420™). Plasma
lipopolysaccharide (LPS)
levels were determined
after 6 weeks treatment
(n=9-10 per group).

24. 24D u P o n t N u t r i t i o n & H e a l t h
Glucoseturnoverrate,
mg/kg.min
Treatment with
Bifidobacterium lactis 420
reverses high-fat diet
(HFD)-induced insulin
resistance. Insulin
sensitivity (glucose
turnover rate in mg/kg
min). (mean±SEM; n=8–
10; data with identical
superscript letters do not
differ with p>0.05).
Amar et al (2011) EMBO Mol Med 3, 559–572
Effect of B. lactis B-420™ on insulin sensitivity

25. D u P o n t N u t r i t i o n & H e a l t h 25
B-420™ ameliorates weight gain
Stenman et al. 2014

26. D u P o n t N u t r i t i o n & H e a l t h 26
Probiotics and prebiotics for controlling body fat mass
• Randomized, double-blind, multicenter study
• Study design and execution follows ICH-GCP
• 225 Finnish participants with BMI 28-34.9, otherwise healthy
• Primary outcome: relative change in body fat mass
Stenman et al. EBioMedicine 2016 ClinicalTrials.gov NCT01978691
Bifidobacterium lactis B-420TM and possible opportunities in
weight management
6Months
Follow-up:
+1 month
Control: microcrystalline cellulose (MCC = 12g/day)
Probiotic: B-420™ (1010 CFU/day) + MCC (12g/day)
Prebiotic: Litesse® Ultra™ polydextrose (12g/day)
Synbiotic: Litesse® Ultra™ (12g/day) + B-420™ (1010 CFU/day)
Baseline

27. D u P o n t N u t r i t i o n & H e a l t h 27
1.4 kg reduction of total Body Fat Mass compared to placebo
Stenman et al. EBioMedicine 2016
Primary outcome: B-420™ + Litesse® Ultra™ control total body fat mass
-4.5% (-1.4 kg)
in fat mass
P=0.02
-4% (-1.4 kg)
In fat mass
Statistical analyses:
ANCOVA values
Factorial analysis

28. D u P o n t N u t r i t i o n & H e a l t h 28
2.6 cm reduction in waist circumference
Stenman et al. EBioMedicine 2016
-2.7% (-2.6 cm)
-2.4% (-2.4 cm)
Statistical analyses:
ANCOVA values
Factorial analysis

29. D u P o n t N u t r i t i o n & H e a l t h 29Stenman et al. EBioMedicine 2016
Evolution of lean body mass during the study (PP)
Month by month increase of lean body mass

30. D u P o n t N u t r i t i o n & H e a l t h 30
B420
Placebo
Akkermansia is more abundant in human subjects consuming B-420™
• B-420™ impacted many species of the human
gut microbiota; including, significantly higher
relative abundance of Akkermansia
muciniphila in B-420™ subjects1.
• Abundance of Akkermansia muciniphila, a
mucin-degrading bacterium that resides in the
mucus layer, is generally associated with
improved metabolic health including body
weight, glucose tolerance and intestinal
permeability2, 3, 4.
• A. muciniphila has been identified as inversely
correlated with body weight gain in pregnant
women.2
• A. muciniphila controlled gut barrier function,
fat mass storage, and glucose tolerance in
obese and type 2 diabetic mice3.
1Hibberd et al. 2018. Beneficial Microbes (under peer review)
2SantaCruz et al. 2010. British Journal of Nutrition
3Everard et al., 2013. Proceedings of the National Academy of Sciences of the United States of America
4Dao et al., 2016. Gut
TM

31. D u P o n t N u t r i t i o n & H e a l t hD u P o n t N u t r i t i o n & H e a l t h 31
Dose-response effect of Bifidobacterium lactis HN019 on whole gut transit time
and functional gastrointestinal symptoms in adults.
Waller et al (2011) Scand J Gastroenterol.
Beneficial effects of Bifidobacterium lactis on lipid profile and cytokines in patients with
metabolic syndrome: A randomized trial. Effects of probiotics on metabolic syndrome.
Bernini et al (2016) Nutrition
Supplementation with Bifidobacterium lactis probiotics in
pregnancy increases levels of TGF-ᵝ and IgA in breast milk
Prescott et al (2008) Clinical and Experimental Allergy
Prebiotic and Probiotic Fortified Milk in Prevention of Morbidities among
Children: Community-Based, Randomized, Double-Blind, Controlled Trial
Sazawal et al (2010) PlusOne
Probiotic with or without Fiber Controls Body Fat Mass, Associated With Serum Zonulin, in Overweight
and Obese Adults. Stenman et al (2016) EBioMedicine.
Bifidobacterium lactis reduces on reduces risk for Diarrhea
and Fever in Preschool Children in an Urban Slum in India.
Hemalatha et al (2014) Eur. J. Nutrition & Food Safety
The Effect of Bifidobacterium animalis HN019 on Cellular Immune Function in
Healthy Elderly Subjects: Systematic Review and Meta-Analysis
Miller et al (2017) Nutrients
Proposed beneficial effects of Bifidobacterium animalis lactis strains

32. D u P o n t N u t r i t i o n & H e a l t h
The Difference a Healthy Microbiome Makes!
• The gut microbiome – changes throughout life, focusing on Bifidobacterium spp.
• Members of the microbiome and their potential role in
− Reducing prevalence of gut infections
− Modulating blood lipids
− Body weight
− Metabolic health
− Immune health later in life
• Dietary tools for making a difference
Overview
32

33. D u P o n t N u t r i t i o n & H e a l t h 33
AIM OF THE STUDY
Determine effects of probiotic intake on microbiota
composition in healthy elderly and the dose response effect.
PRODUCT FORMULA
3 doses of Bifidobacterium lactis HN019™ in 250 ml of milk
• 5 x 109 CFU (high)
• 1 x 109 CFU (medium)
• 6.5 x 107 CFU (low)
• Placebo (reconstituted skim milk)
SUBJECTS
20 healthy elderly (>60 years) in each group.
STUDY DESIGN
• Randomized, double-blind, placebo controlled study.
• 2 week run-in, 4 week intervention, 2-week wash-out
• Bacterial population levels detected in fecal samples at 6
time points
PUBLICATION
Ahmed at al. J Nutr Health Aging. 2007 Jan-Feb;11(1):26-31
https://www.ncbi.nlm.nih.gov/pubmed/17315077
Oral administration of Bifidobacterium lactis HN019™
B. lactis HN019™ increases the amount of beneficial bifidobacteria in the
microbiota of elderly
 In all HN019™ dose groups, bifidobacterium
population increased significantly during
intervention when compared to pre-
intervention levels.
 Similarly lactobacilli population increased
in high and medium dose groups.
 No changes in placebo group.
Bifidobacteria levels
* P< 0.005
33

34. D u P o n t N u t r i t i o n & H e a l t h 34
Std diff in means + 95% CI*
Bifidobacterium lactis HN019™
HN019™ improves
phagocytic capacity
HN019™ improves
NK cell activity
Total Total
*Standard difference in means + 95% confidence interval
Miller, Lehtoranta, and Lehtinen 2017 Nutrients. http://www.mdpi.com/2072-6643/9/3/191
Std diff in means + 95% CI*
Daily consumption of B. lactis HN019™ enhances
immune function in healthy elderly adults
TM
TM
34

35. D u P o n t N u t r i t i o n & H e a l t hD u P o n t N u t r i t i o n & H e a l t h 35
Dose-response effect of Bifidobacterium lactis HN019 on whole gut transit time
and functional gastrointestinal symptoms in adults.
Waller et al (2011) Scand J Gastroenterol.
Beneficial effects of Bifidobacterium lactis on lipid profile and cytokines in patients with
metabolic syndrome: A randomized trial. Effects of probiotics on metabolic syndrome.
Bernini et al (2016) Nutrition
Supplementation with Bifidobacterium lactis probiotics in
pregnancy increases levels of TGF-ᵝ and IgA in breast milk
Prescott et al (2008) Clinical and Experimental Allergy
Prebiotic and Probiotic Fortified Milk in Prevention of Morbidities among
Children: Community-Based, Randomized, Double-Blind, Controlled Trial
Sazawal et al (2010) PlusOne
Probiotic with or without Fiber Controls Body Fat Mass, Associated With Serum Zonulin, in Overweight
and Obese Adults. Stenman et al (2016) EBioMedicine.
Bifidobacterium lactis reduces on reduces risk for Diarrhea
and Fever in Preschool Children in an Urban Slum in India.
Hemalatha et al (2014) Eur. J. Nutrition & Food Safety
The Effect of Bifidobacterium animalis HN019 on Cellular Immune Function in
Healthy Elderly Subjects: Systematic Review and Meta-Analysis
Miller et al (2017) Nutrients
Proposed beneficial effects of Bifidobacterium animalis lactis strains

36. Thank You
©2019 DowDuPont, The Dow Chemical Company, DuPont. All rights reserved.
®™ Trademark of DowDuPont, The Dow Chemical Company, E.I. du Pont de Nemours and Company, or their affiliates.
DuPont Nutrition & Health, a DowDuPont Specialty Products Division business, combines in-depth knowledge of food and nutrition
with current research and expert science to deliver unmatched value to the food, beverage, pharmaceutical and dietary supplement
industries. We are innovative solvers, drawing on deep consumer insights and a broad product portfolio to help our customers turn
challenges into high-value business opportunities. More information is available at www.dupontnutritionandhealth.com.