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Coated calcium butyrate in calf milk replacers: mode of action, application and benefits Dr. Noémie Van Noten – Sanluc International


Presentation overview 1. Introduction on butyrate 2. Mode of action 3. Benefits 4. Application 5. Experimental part 6. Conclusions


1. Introduction

Endogenous butyrate production Butyric acid = product of bacterial fermentation of dietary fiber: o in the rumen of ruminants o in the hindgut of monogastrics and ruminants

Cui et al. (2019). Compr. Rev. Food Sci. Food Saf. 18: 1514-1532.


1. Introduction

Butyrate supplementation o Milk fat contains ± 4-5% butyric acid o But, calf milk replacer: Ø Vegetal or animal fat Ø Low butyric acid content

Calcium

o Butyric acid: Ø Volatile Ø Unpleasant odour Ø Liquid

→ Supplementation as (coated) Ca-salt (CH3-CH2-CH2-COO)2-Ca


2. Mode of Action

Molecular Mechanisms Energy source

Butyric acid

Signaling molecule: GPRs

Epigenetic regulation: HDAC


2. Mode of Action

Butyrate as energy source ATP

Colonocyte

o Butyrate = preferred energy source of colonocytes o Only small fraction of luminal butyrate enters the blood

Portal vein

circulation o Uptake in liver almost complete: energy

Liver


2. Mode of Action

Molecular Mechanisms Energy source

Butyric acid

Signaling molecule: GPRs

Epigenetic regulation: HDAC


2. Mode of Action

Butyrate as signaling molecule Butyrate interacts with G protein-coupled receptors (GPRs) = transmembrane proteins that detect molecules outside the cell and activate cellular responses Involved in: • sensing luminal contents • intestinal motility • secretion of hormones • innate immunity

Liu et al. (2018). Adv. Nutr. 9:21-29


2. Mode of Action

Molecular Mechanisms Energy source

Butyric acid

Signaling molecule: GPRs

Epigenetic regulation: HDAC


2. Mode of Action

Butyrate as epigenetic regulator Histone Deacetylases (HDACs) o Enzymes o Remove acetyl groups from peptide chains o Affect mainly histones (DNA-packing) o DNA becomes condensed and silenced

Butyrate = HDAC inhibitor o Histone tails remain acetylated → Genes remain active → Affects a large amount of genes → Essential for gut homeostasis Stilling et al. (2016), Neurochemistry International 99, 110-132


3. Benefits Molecular Mechanisms

Triple mode of action

Energy source

Butyric acid Signaling molecule: GPRs

Epigenetic regulation: HDAC

Gut Physiology and Digestion

Inflammation control

Butyric acid

Microbiota Modulation


3. Benefits

Gut Physiology and Digestion o Cell proliferation ↑, apoptosis ↓ Ø Longer intestinal villi Ø Rumen papillae development

o Activity of digestive enzymes↑:

Control

Butyric acid

Ø Brush border enzymes Ø Pancreatic enzymes

o Improved intestinal barrier function

Improved digestion and absorption Mentschel et al. (2001), Archives of Animal Nutrition, 55(2), 85-102.


3. Benefits Molecular Mechanisms

Triple mode of action

Energy source

Butyric acid Signaling molecule: GPRs

Epigenetic regulation: HDAC

Gut Physiology and Digestion

Inflammation control

Butyric acid

Microbiota Modulation


3. Benefits

Inflammation control and immunomodulation o Inflammation reduction Ø Regulation of pro- and anti-inflammatory mediators (IL, PPAR-γ ) Ø Increased heat shock protein expression Ø Upregulation of antioxidant enzymes

o Immune regulation Ø Less excessive innate immunity Ø More specific immune reactions

Disease resistance ↑ & nutrients for growth↑ Parada Venegas et al. (2019). Frontiers in immunology, 277.


3. Benefits Molecular Mechanisms

Triple mode of action

Energy source

Butyric acid Signaling molecule: GPRs

Epigenetic regulation: HDAC

Gut Physiology and Digestion

Inflammation control

Butyric acid

Microbiota Modulation


3. Benefits

Modulation of Microbiota o Mutual relationship Ø Microbiota composition ↔ butyric acid concentration Ø Firmicutes / Bacteroidetes-ratio ↑

o Pathogen control Ø Directly or indirectly Ø Salmonella, Clostridium perfringens, E. coli


3. Benefits

Improved PERFORMANCE

Triple mode of action Gut Physiology and Digestion

Inflammation control

Butyric acid

Microbiota Modulation


4. Application

Digestion of newborn calf Young calf = pre-ruminant o Milk bypass via esophageal groove o Rumen, reticulum and omasum do not work o Enzymatic milk digestion in abomasum o Rumen development : Ingestion of solid feed


4. Application

Butyrate Supplementation Rearing calves

Veal + Rearing calves

Solid Feed

Milk Replacer

Uncoated CaBut

Coated CaBut

Uncoated CaBut

Coated CaBut

Release: abomasum Effect: Abomasum pancreas prox. SI

Release: gradual Effect: Abomasum pancreas prox. SI distal SI

Release: rumen Effect: Rumen

Release: gradual Effect: Forestomach Abomasum prox. SI


5. Calf trials 1. Intestinal morphology 2. Veal calves – Germany 3. Veal calves - Belgium


5. Experimental part: trial 1

Gut morphology calf trial Protocol: o o o o

50 calves with average age of 94 days Pre-trial: 3 days on conventional milk powder Trial diets: 12 days Dietary treatments: Ø T1: Conventional milk replacer (CMR) Ø T2: Modified milk replacer (MMR) with 8% soy protein Ø T3: MMR + 1,28 kg/ton coated calcium butyrate (CCB) Ø T4: MMR + 2,56 kg/ton CCB Ø T5: MMR + 5,12 kg/ton CCB


5. Experimental part: trial 1

Small intestinal morphology Jejunum Villus height (µm)

2000 1500

1305

❷

❶ 1131

1241

1380

1511

MMR

1000

MMR+1,28CCB MMR+2,56CCB

500 0

CMR

MMR+5,12CCB b

c

bc

ab

a

❶ MR with soy protein significantly reduces intestinal villi height ❷ CCB increases intestinal villi height in a dose dependent manner


5. Experimental part: trial 1

Small intestinal morphology

CMR MMR + 1,28 CCB

MMR MMR + 2,56 CCB

MMR + 5,12 CCB


5. Experimental part: trial 2

Calf trial - Germany Protocol: o 30 male calves (German Holstein); housed individually o Trial period: Ø d9-d11 of age: adaptation period Ø d12-d68 of age: trial period (56 days) o Feeding regime: calf milk replacer + calf starter + straw o Dietary treatments: Ø Control group: basal milk replacer Ø CCB group: basal milk replacer + 2,58 kg/ton CCB


5. Experimental part: trial 2

Calf trial - Germany 100

Body weight (kg)

Results:

+2,3%

70,8

80 60

50,8

94,3

96,5

b

a

72,8

50,6

40 20 0 d0

d21 Control

d56

CCB

o No differences in feed intake o Significant improvement of FCR: Ø Control: 1,927 -4,5% Ø CCB: 1,840


5. Experimental part: trial 3

Calf trial - Belgium Protocol: o o o o

50 male calves (Holstein Friesian); housed individually Age at arrival: between 10 and 14d Trial period: 56 days Feeding regime: Ø d0-d18: calf milk replacer + calf flakes Ø d19-56: calf milk replacer + concentrate enriched with roughage

o Dietary treatments: Ø Control group: basal milk replacer Ø CCB group: basal milk replacer + 2,50 kg/ton CCB


5. Experimental part: trial 3

Calf trial - Belgium

+2,2%

Results:

88,3 90,2

80 60

45,5 45,5

49,6 49,6

d0

d14

60,2 61,1

72,5 73,5

40 20 0 Control

d28 CCB

d42

1000 800

d56

1126 1191

1200

ADG (g/d)

Body weight (kg)

100

760 824

877 884

d14-28

d28-42

+4,5% 764 798

600 400

291 292

200 0 d0-d14

d42-56

d0-56


5. Experimental part: trial 3

Calf trial - Belgium

+2,9%

1000

Results: ADFI (g)

800

741

752

741

774

600 400 200 0 Milk powder Control

Concentrate CCB

o Significant improvement of FCR: Ø Control: 1,953 -3,2% Ø CCB: 1,890


6. Conclusions

Coated calcium butyrate in calves o Dose recommendation: Ø Milk powder: 1 - 3 kg/ton Ø Solid starter feed: 1,5 - 2 kg/ton

o Benefits: Ø Ø Ø Ø Ø

Improved intestinal, morphology and health Less diarrhoea and intestinal problems Improved weight gain and FCR Extensive development of rumen papillae Faster and higher prestarter feed intake

Early development of a functional GIT resulting in high performing and healthy calves


Coated calcium butyrate in calf milk replacers: mode of action, application and benefits Thank you for your attention!


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