August 24, 2015

Developing a gut health solution

requires a new holistic approach

Arie K. Kies

Page

Why are we interested?

• DSM: technical solid company; lots of nutrition knowledge

• We want to strengthen support to customers further

Providing solutions according to their needs

Through bright science

Page

• Reducing use of antibiotics (improving gut health)

• …

Problems of customers?

Focus within poultry-feed industry:

Page

State of affairs• Efficacy AGP

• Risks

• Effect of termination

• Research (opportunities)– Genomics

– Immune modulation

• Alternatives– Pre- and probiotics

– Acids

– Enzymes

– Botanicals

– Bacteriophages

2005

Page

State of affairs

2015

• Research – Genomics

– Immune modulation

– Microbiomics

• Alternatives– Pre- and probiotics

– Acids

– Enzymes

– Botanicals

– Bacteriophages?

Page

State of affairs

• Little progress – Reducing AGP’s

– Improving gut health

• Do we look to the right things?

A different angle is required

Page

Approach so far

“AGP alternatives are being sought to improve health,

immune status and performance in animal agriculture.”

“Most AGP alternatives try to prevent proliferation of

pathogenic bacteria and modulate indigenous bacteria

so that the health, immune status and performance are

improved.”

“While the industry is focused on developing solutions that

work similarly, AGPs' exact mode of action is not yet fully

understood.”

Mallo, Feed International, 2015

Page

Approach so far

“AGP alternatives are being sought to improve health,

immune status and performance in animal agriculture.”

“Most AGP alternatives try to prevent proliferation of

pathogenic bacteria and modulate indigenous bacteria

so that the health, immune status and performance are

improved.”

“While the industry is focused on developing solutions that

work similarly, AGPs' exact mode of action is not yet fully

understood.”

Mallo, Feed International, 2015

Why?

Page

Approach so far

“AGP alternatives are being sought to improve health,

immune status and performance in animal agriculture.”

“Most AGP alternatives try to prevent proliferation of

pathogenic bacteria and modulate indigenous bacteria

so that the health, immune status and performance are

improved.”

“While the industry is focused on developing solutions that

work similarly, AGPs' exact mode of action is not yet fully

understood.”

Mallo, Feed International, 2015

Why?

Possible?

Page

Approach so far

“AGP alternatives are being sought to improve health,

immune status and performance in animal agriculture.”

“Most AGP alternatives try to prevent proliferation of

pathogenic bacteria and modulate indigenous bacteria

so that the health, immune status and performance are

improved.”

“While the industry is focused on developing solutions that

work similarly, AGPs' exact mode of action is not yet fully

understood.”

Mallo, Feed International, 2015

Possible?

Page

Mode of action of AGP’s?

• How to explain consistent positive effect of AGP’s?• Still consistent in modern diets?

• Do we know what we are looking for??

Change in – microbial population?

– microbial activity?

– development intestine?

• AGP’s are not strain-specific

• AGP’s are usually dosed below MIC

Page

Mode of action of AGP’s?

AGP’s are not strain-specific

Example Avilamycin (MIC90-values)

Clostridium spp. (+) 8 (µg/ml)

Bifidobacterium spp. (+) 32

Lactobacillus spp. (+) > 128

E. Coli (-) > 128

Bacteroides fragilis (-) 32 (normally commensal)

Enterococcus spp. (+) 4 (lactobacilli-order)

FDA, 2015

Page

Mode of action of AGP’s?

AGP’s usually dosed below MIC

Again Avilamycin as example

- Max. dose allowed in feed was 10 ppm

- Assume 5-10* dilution in small intestine

concentration (max.) 1-2 µg/ml

[MIC90 Clostridiae: 8 µg/ml]

Specificity and MIC90-values alternatives?

Place (in g.i.t.) of activity?

Page

Mode of action of AGP’s?

Koopman, 1999

A balanced, stable microflora protects against pathogensNumber Salmonellae/g fresh feces

Problem not pathogens but dysbiosis

- Germfree mice

- Measured with culturing

Page

Gastrointestinal Microbes of chickens

Schokker, unpublished

Microflora is dynamic. Effect of age:

Enterobacteriaceae (Escherichia, …)

Enterococceae (Lactobacilli)

Clostridiaceae

Lactobacilaceae

Streptococcaceae

Page

Gastrointestinal Microbes of chickens

Schokker, unpublished

Microflora is dynamic. Effect of age:

Enterobacteriaceae (Escherichia, …)

Enterococceae (Lactobacilli)

Clostridiaceae

Lactobacilaceae

Streptococcaceae

Need to develop balanced gut ecosystem

May vary in time!

Page

Gut health solution

Improving gut health more consistently:

- Last 10 years products developed that aid overcoming

the “AGP-gap”

- Lack of consistent improvement in many products

- Wet litter is still a recurrent issue

- Despite a lot of (scientific) work

- Too little progress made

- (Too) little interdisciplinary research

Page

Gut health solution

Improving gut health more consistently:

How to go on?

Page

Gut health solution

Improving gut health more consistently:

Holistic approach

Gut health: - A complex system

- A complex problem

- Many different angles; different competences

Input from different disciplines required!

Page

Gut health solution

A complex system -- interactions within the gut

Nutrition

Management, Environment, Genotype

Excretion

Nutrient utilization Immune competence

Performance

gutnutrition genotype

microbiota

From: Smits, 2013

Page

Gut health solution

Improving gut health more consistently:

Holistic approach (farm perspective)

• Farm management

• Genetics

• Quality day-old chicks

• Feed- Feeding schedule

- Quantity

- Form (particle size; mash vs. pellets)

- Composition (feedstuffs, level)

- Additives

• …

Specific solution required per farm / company!

Page

Gut health solution

Form (particle size; mash vs. pellets)

But: caecal microbial diversity lower …

Day 0 - 34 Fine Coarse

Feed Intake (g/b) 2991 3144

BW gain (g/b) 1730 1877

Water intake (g/b) 5445 5305

Gizzard weight (g/100 g BW) 1.36 1.60

Villus height (µm) 1517 1840

Crypt depth (µm) 292 230

Qaisrani et al., Poultry Sci. 2014; Qaisrani, 2014

RSM-based diet (‘badly’ digestible protein; different levels)

2 studies

Page

Gut health solution

Additives

• “… Vitamin A in the womb determines … life-long immunity”

• “… immune health in adulthood can be preset by nutritional status

during fetal life”

Bird, Nature Rev. Immunol., 2014

Page

Gut health solution

Additives

Brown et al., Immunity, 2015

“- Retinoic acid (RA) stabilizes Th1 fate commitment

- RA-RAR prevents development of pathogenic Th17 cells in vivo”

“Good”

“Bad”

Page

Gut health solution

Improving gut health more consistently:

Holistic approach (scientific disciplines perspective)

• Animal nutrition

• Human nutrition

• Veterinary science

• Human medical sciences

Page

Human medical sciences?

- Clostridium perfringens Necrotic enteritis

- Aetiology similar to C. difficile in humans• Organism itself: non-invasive

• Since 2000 dramatic increase severe cases (2/3 hospital acquired)

– “Most important risk factor for C. difficile infection remains antibiotic

use” [!!]

– “Minimizing antibiotics use reduced infection rate by 77% (in a 450-bed

hospital, Scotland)”

– Treatment: antibiotics

– Prevention probiotics? “… not recommended [at this moment]”, but:

“fecal microbiota transplantation seems very promising”

Leffler and Lamont, NEJM 2015

PageVan Nood et al., NEJM 2013

Human medical sciences?

Fecal microbiota transplantation

Rates of cure:

Patients without relapse

for recurrent Clostridium

difficile infection

Also microbiota diversity

increased dramatically by

fecal transplantation

Page

Gut health solution

Improving gut health more consistently:

Holistic approach (scientific perspective)

• Nutrition– Protein, energy, …

• Feed technology– Milling, pelleting, extrusion, …

• Physiology– Digestion, utilization, …

• Immunology– Immune competence, inflammation, …

• Microbiology– Strains, numbers, …

• “Omics”– Genomics, microbiomics, metabolomics, …

Page

Gut health solution

Practice: reducing protein often effective

But protein is required

Wijtten et al., Poultry Sci., 2004

♂

♀

CP: ~21 - ~27%

Page

Gut health solution

Practice: reducing protein often effective

Fermentation of undigested protein

Qaisrani, 2014

Page

Gut health solution

Practice: reducing protein often effective

Fermentation of undigested protein

Metabolites– Biogenic amines

– Branched chain fatty acids

– Phenols, thiols, ammonia, indoles, …

– Nitrate

• Are they negative by definition?

• Interaction with fermentable carbohydrates

M’Sadeq et al., An. Nutr., 2015

Page

Metabolomics: big data

>200.000 features for negative scan mode

and

>200.000 for positive, total ± 450.000

Filter 1: ± 80.000 left

Filter 2

Metabolites

identified:

1000

Mass spectrometry Bioinformatics pipeline

MetabolitesAnim

als

on d

iffe

rent

die

ts

Correlations between diet,

intestinal phenotype and

metabolites/microbial species

Page

Program with different university-groups

- Animal nutrition

- Human medical research

- Veterinary

- Metabolomics

- Microbiology

- …

Understanding animal gut health; emphasis on undigested, fermented

protein, in relation to energy

Impact microbial activity on dysbiosis

Influence of nutritional interactions (feed composition, quality of

ingredients, technology, additives, …)

Gut health solution – our holistic approach

Page

The DSM gut health solution

Created for customers,

driving by science