The science behind peptidoglycans - Pietro Celi.pdf · of both gram- positive (G+) and...
Transcript of The science behind peptidoglycans - Pietro Celi.pdf · of both gram- positive (G+) and...
For Internal Use Only
The science
behind
peptidoglycans
Pietro Celi (DVM, PhD)
Global Science Lead Eubiotics
DSM Nutritional Products, Animal Nutrition and Health
For Internal Use Only
PRESENTATION OUTLINE
Gastrointestinal functionality
The problem Peptidoglycans
Can we fix it?Muramidase and gastrointestinal
functionality
Take home messages
GASTROINTESTINAL FUNCTIONALITY#FEED4GAIN
For Internal Use Only
#FEED4GAIN
GASTRO-INTESTINAL (GAIN) FUNCTIONALITY
‘a steady state where the
microbiome and the intestinal
tract (host) exist in symbiotic
equilibrium and where the
welfare and performance of the
animal is not constrained by
intestinal dysfunction’
Welfare
Diet
GIT
microbiota
Digestion &
absorption
GIT mucosa
Immune
status
Gastrointestinal
Functionality
For Internal Use Only
Animal performance
Antimicrobial resistance
(#AMR)
#Welfare & Environment
#Food Safety
#sustainableanimal
production
WHY GASTROINTESTINAL FUNCTIONALITY?
Gastrointestinal Functionality is the single most important factor
impacting poultry producers profitability and license to produce
THE PROBLEMWASTE IN THE GUT
For Internal Use Only
Intensification
Stocking density
Removal of AGPs
Incidence of intestinal problems
CHALLENGES IN THE POULTRY INDUSTRY
For Internal Use Only
INTESTINAL CHALLENGES
GASTROINTESTINAL FUNCTIONALITY
Necrotic Enteritis & coccidiosis
Salmonella & Campylobacter
Malabsorption
Bone mineralization
(skeletal health)
For Internal Use Only
GASTROINTESTINAL FUNCTIONALITY
Welfare
Maximise value from
feed
Robust immune system
Nutrient digestibility
Management practices
Is there something new out there?
WHAT IS NEEDED
THE GREATEST THREAT IS THE ONE THAT GOES UNOBSERVEDPEPTIDOGLYCANS
For Internal Use Only
GIT MICROBIOTA: DEAD OR ALIVE?
UNLOCKING THE HIDDEN POTENTIAL
Gadde, U., Lillehoj, H. 2017. Antibiotic Alternatives- A Review. Animal Health Research Reviews.
Millions of microbes are swallowed continuously
Billions of microbes are present in the GIT tract
Microbes are continuously dying off
What happens to the dead bacteria?
Completely overlooked area= bacterial cell debris
For Internal Use Only
ALL PARTS INTERACT WITH THE HOST CELLS IN THE GIT
A HOLISTIC VIEW OF MICROBIOTA INCLUDES DEAD BACTERIA AND CELL DEBRIS
Dead bacteria
(whole cells + fragments)
Live bacteria +
For Internal Use Only
PEPTIDOGLYCANS ARE MAJOR BACTERIAL CELL WALL COMPONENTS
Gram-bacterium
Gram+bacterium
For Internal Use Only
When bacteria and their cell debris accumulate on the intestinal mucosa
• impair absorption
• enterocyte damage
• increased intestinal permeability
PEPTIDOGLYCANS IN THE GUT HINDER GASTROINTESTINAL FUNCTIONALITY
For Internal Use Only
GUT MICROBIOTA LOAD
increase in the GIT microbiota
load
increase in bacterial cell
debris
more PGNs
Source: Maresca, 2013
Dietary protein content
Starch content
NSP levels
Antinutritional factors
Micotoxins
Environmental stress
Pathogen load
MORE BUGS, MORE PEPTIDOGLYCANS
For Internal Use Only
GIT microbiota is composed of both gram- positive (G+)
and gram-negative (G-) bacteria.
When cell walls accumulate in the GIT tract they
become “intestinal waste”
Nutrient digestion and absorption are particularly
prone to the adverse impact this of this cell
debris.
SUMMARY
GASTROINTESTINAL FUNCTIONALITY
Bacterial Overgrowth Impacts Nutrient Absorption
CAN WE FIX IT?YES WE CAN
For Internal Use Only
DISCOVERED AND DEVELOPED BY THE DSM/NZ ALLIANCE
THE DISCOVERY OF MURAMIDASE 007
Expression
Diversity
selection
In silico
In vivo
FCR
Purification Production feasibility
Stability tests
Counter screen
Antimicrobial
potency
Activity screen
Peptidoglycan hydrolysis
Idea review
Project kick- off
Upscale and formulation
ProductLead
enzyme
Iteration cycles
Muramidase 007 was selected as the best out of >
1´000 muramidase candidates
For Internal Use Only
NOT ALL MURAMIDASES ARE THE SAME
THE SOLUTION TO THE PROBLEM
Muramidase 007 specifically catalyze the hydrolysis
of peptidoglycans
MURAMIDASE 007HEWL
For Internal Use Only
NOVEL MURAMIDASE
THE SOLUTION TO THE PROBLEM
Activity on cell debris peptidoglycan component
No antimicrobial potency
Optimal pH activity range 4-7.5
Stable under gastric conditions
Heat stable
For Internal Use Only
BEFORE AND AFTER MURAMIDASE INCUBATION
PEPTIDOGLYCANS
For Internal Use Only
Removing bacterial cell debris promotes optimal digestion and absorption
Releasing amino acids and sugars for host and microbiota
Less damage to the intestinal mucosa
• less energy for tissue repair
• more energy for production purposes
HYDROLYZING PEPTIDOGLYCANS MAY RELEASE A HIDDEN POTENTIAL
UNLOCKING THE HIDDEN POTENTIAL
A hidden potential inside the gastrointestinal
tract = Bacterial Cell Debris
MURAMIDASEIMPROVES GASTROINTESTINAL FUNCTIONALITY#FEED4GAIN
For Internal Use Only
MURAMIDASE INCREASES ABSORPTION CAPACITY
Male Ross 308 fed with diets based on SBM/ Corn/ Rice bran (5%)/ MBM + HiPhos + Carophyll Yellow Challenge: 15x RD Bio-Coccivet R®
*AGP = Enramycin (10ppm Starter and Grower + 5ppm Finisher)
a b ab b
L. Cardoso, 2016
Measurement with iCheck™ Carotene (quick test).
Bittencourt et al., 2018
For Internal Use Only
FITC-Dextran
Lower FITC in blood =
better gut permeability
Cardoso, 2017
FITC-DEXTRAN AS A MARKER FOR GUT PERMEABILITY: NUTRIENT
ABSORPTION AND BACTERIA TRANSLOCATION
MURAMIDASE AND NUTRIENT ABSORPTION
For Internal Use Only
Source: BR 170330 (UFPR) - A. Maiorka, et al., 2017. Unpublished data
MURAMIDASE IMPROVES INTESTINAL PERMEABILITY AND ABSORPTION CAPACITY
For Internal Use Only
Source: ME-01/17VN Aureli & Perez Calvo. Unpublished data
INTESTINAL HISTOMORPHOLOGY: VILLI LENGTH AND CRYPT DEPTH, AND
MUCOSA THICKNESS
MURAMIDASE DECREASES MUCOSA THICKNESS
For Internal Use Only
WELFARE
MURAMIDASE DECREASES WET LITTER & FPD
84,9
97,9
14
2,121,07
75
80
85
90
95
100
NC BOND
FPD score (%) day 36
Score 2
Score 1
Score 0
a b
FPD score = 9.12 a 1.08bEsteve & Perez Calvo, 2019
2
CTR M007
CTR M007
For Internal Use Only
Increases Weight Gain (3%)
Improves digestibility of nutrients
Reduces thickness of GIT mucosa
Increases breast meat yield
Less wet litter and FPD
Improves FCR (-3%)
Increase absorption capacity
MURAMIDASE IMPROVES GASTROINTESTINAL FUNCTIONALITY
Muramidase
For Internal Use Only
There are a lot of birds out there
with their gut full of peptidoglycans
What are you going to do about it?
#FEED4GAIN
TAKE HOME MESSAGES
BRIGHT SCIENCE. BRIGHTER LIVING.
For Internal Use Only
#FEED4GAIN
TAKE HOME MESSAGES
Game Changing Innovation
Unlocking the Hidden
Potential
Novel and Unique
Muramidase
PGNs impact nutrient
absorption
Supporting gastrointestinal
functionality
Sustainable and environmentally friendly solution