This document discusses probiotics and probiotic research. It begins with a historical overview of Élie Metchnikoff's early 20th century work suggesting that modifying gut flora with beneficial microbes could promote health and longevity. It then defines probiotics and provides examples of common probiotic strains. Several studies are summarized that show the potential health benefits of various probiotic strains and formulations in managing gastrointestinal symptoms and antibiotic-associated diarrhea. The document also discusses factors that influence probiotic survival like formulation, delivery systems, and storage conditions. It concludes with an overview of DuPont's probiotic expertise and product portfolio.

1. 1
To take probiotics, or
not to take probiotics?
4 May 2018
Anders Henriksson, PhD
Principal Application Specialist
DuPont Nutrition & Health, Australia/ New Zealand

2. Questions
2
Do probiotics survive
transit trough the
stomach?
Do different probiotics
compete in a capsule?
What is a probiotic strain?
Will antibiotics kill probiotics if
they are taken simultaneously?
Are multi-strain probiotics more
effective then single strain
probiotics?
What makes a
probiotic shelf stable?
Have these probiotics
been encapsulated?
How are probiotics produced?
Give some
examples of how do
probiotics work?

3. Lactic acid bacteria and their impact on health -
the views of Élie Metchnikoff
Russian scientist and Nobel laureate Élie Metchnikoff, who in the beginning of the 20th
century suggested that it would be possible to modify the gut flora and to replace harmful
microbes with useful microbes.
Metchnikoff had observed that certain rural populations in Europe, for example in Bulgaria and
the Russian steppes who lived largely on milk fermented by lactic-acid bacteria were
exceptionally long lived.
E. Metchnikoff, 1907

4. What are probiotics?
Probiotics are ”Live microbes that, when
administered in adequate amounts, confer a
health benefit on the host”
FAO/WHO 2002; Hill et al., 2014
Bifidobacterium longum
Lactobacillus acidophilus
Lactobacillus crispatus
Lactobacillus reuteri Lactobacillus casei
Lactobacillus paracasei
Lactobacillus rhamnosus
Bifidobacterium animalis lactis
Bifidobacterium infantis

5. Meta-studies evaluating the efficacy of single vs. multi-species/strain probiotics
5
Authors’ conclusions:
“Based on a limited number of
studies, multi-strain probiotics
appear to show greater efficacy
than single strains, including
strains that are components of the
mixtures themselves”

6. The effect of Bifidobacterium lactis HN019™ on gastrointestinal
symptoms in adults
Daily dosage levels tested:
1.8 x109 CFU
1.7 x1010 CFU
6

7. 7
Effect of B. lactis HN019™ on gastrointestinal symptoms in healthy adults
N=34
N=100
N=33
Probiotic 1.7 x 1010 CFU/d
Placebo feeding
Probiotic 1.8 x 109 CFU/dN=34
Day 0 Day 14
*
*
*
*
Digestive symptoms measured included:
▪ Constipation, abdominal pain, irregular bowel movements,
▪ Scale - [1 (never) to 4 (always)]
100 subjects (age: 44 yr, 64% female)

8. 8
Changes in GTT in healthy adults after 14 days consumption of B. lactis HN019™
-10
-5
0
5
10
15
20
25
30
35
40
1 2 3
ReductioninCTT(hrs)
17B/day 1.8B/day Placebo

9. Probiotics in management of on Antibiotic Associated Diarrhea (AAD) in adults
9
AIM OF THE STUDY
This study was designed to determine the dose response effect of HOWARU®
Restore formulation on the incidence of AAD and CDAD and severity of
gastrointestinal symptoms in adults having antibiotherapy.
STUDY DESIGN
Triple blind, randomised, placebo controled
SUBJECTS
Adults in-patients requiring antibiotherapy.
450 PATIENTS DIVIDED IN 3 TREATMENT GROUPS COMPLETED THE STUDY
• Placebo
• Combination of 4 probiotic strains at 4.17 x 109 CFU/day (low dose)
• Combination of the same 4 strains at 1.70 x 1010 CFU/day (high dose)

10. Probiotics in management of on Antibiotic Associated Diarrhea (AAD) in adults
- incidence
10
24,6%
19,6%
12,5%
Placebo Low dose High dose
*
Subjects taking the high dose experienced a 50% reduction in incidence of AAD
compared to the placebo group (p<0.005).
Low dose group has shown a 20% reduction compared to placebo group but the results
were not statistically significant.
Ouwehand et al (2014) Vaccine vol 32

11. • Technology was derived from University of Wisconsin patent application entitled “Preservation and
Storage Medium for Biological Materials” #6,653,062
• Technology exclusively licensed to DuPont for bacterial applications
Maintain
membrane
integrity
Phospholipid bilayer cell membrane
Freeze-Drying
= Polar Head-group on Lipid Bi-layer
= Water Molecule
= Saccharide/Salt Polymer
Head-group spacing lost
and channels formed
Membrane
becomes
permeable
STABILIZATION EXPERTISE | Maximize cell membrane integrity during drying
and storage
11

12. 1E+08
1E+09
1E+10
1E+11
0 3 6 9 12 15 18 21 24
CFUpersachet
Time (months)
25°C/60% RH 30°C/75% RH
Bifidobacterium lactis HN019™ (2B)
Lot # 20150529 – S1
Lot number:
20150529 – S1
Excipient:
Maltodextrin, Inulin
and Calcium Silicate
T zero Aw:
0.059
Format:
Powder
Packaging:
Foil sachet
Study start:
June 2015
Label claim 2B
per sachet
12

13. The effect of delivery vehicle on survival of probiotics in gastric juice
13Tee et al, unpublished
TM

14. DuPont Nutrition & Health combines in-depth knowledge of food and nutrition with current
research and expert science to deliver unmatched value to the food, beverage 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
14
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