enhancing calf gut health · dietary regimes in early life influence lifetime productivity 1kg of...

Enhancing Calf Gut Health

Michael A. Steele, Department of Animal Biosciences

I. Calf Management Trends

II. Colostrum

III. Plane of Nutrition

IV. Weaning

V. Antimicrobial (SCB Story)

Enhancing Calf Gut Health

(Adapted from Conrad’s Waddington epigenetic landscape)

“…early adaptation to a stress or stimuli that permanently changes the physiology and metabolism of the organism and continues to be expressed even in the absence of the stimulus/stress that initiated them…”

(Patel and Srinivansan, 2002)

“Nutritional Programming”

Dietary regimes in early life influence lifetime productivity

1kg of pre-weaning ADG = 1,540 kgs of milk in first lactation

(Soberon et al., 2012)

Early Life Nutrition

Gut Health and Dairy Calves

Mortality and Morbidity:

5% mortality, 32% due to digestive disorders

Mean age: 18.3 ± 2.3 d old

12.1% of calves failed passive transfer

26.8% of calves receive antibiotics

48.4% for digestive disorders

Immune Status:

Antibiotic Use:

(Shivley et al. 2018)

(Urie et al. 2018)

38% morbidity, 56% due to digestive disorders

Pre and Post-WeaningPre-ruminant Ruminant

Milk Solid Feed

Weaning Transition

1 wk 4 wk 8 wk 12 wk

Pre and Post-WeaningPre-ruminant Ruminant

Milk Solid Feed

Weaning Transition

1 wk 4 wk 8 wk 12 wk

Mixture of absorptive, goblet, paneth and neuroendocrine cells

Microbial richness and diversity increases through the lower gut

Cell junction proteins are expressed differentially (Malmuthuge et al., 2012)

Gut Epithelium

(Steele et al., 2016)

104/mL 1010/mL1010/mL

Industry Concerns

AntimicrobialMaternal

Colostrum Plane of Nutrition

Knowledge Gaps

Industry Concerns

AntimicrobialMaternal

Colostrum Plane of Nutrition

Knowledge Gaps

(Sharifi et al., 2009)

Bottle Tube

Colostrum Feeding Method

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nce

ntr

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on

(m

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Time Relative to Colostrum Feeding (minutes)

Acetaminophen

Bottle

Tube

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Me

an

IgG

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nc.

(m

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Time Relative to Colostrum Feeding (minutes)

BottleTube

IgG

Colostrum Feeding Method

(Desjardins-Morrissette et al., 2018)

Delayed Colostrum Feeding

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5

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0 h 6 h 12 h

(Fischer et al., 2018)

IgG

mg/ml

Hours after birth

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2.0

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Pro

po

rtio

n (

%)

Treatment

Bifidobacteria associated with colon mucosa

a

ab

b

0.00.10.20.30.4

0h 6h 12h

Pro

po

rtio

n (

%)

Treatment

Lactobacillus associated with colon mucosa

aab

b

Delaying the first colostrum meal may delay the colonization of

beneficial bacteria to the calf intestine

(Fischer et al., 2018)

Delayed Colostrum Feeding

Heat Treatment of Colostrum

0

0.1

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NC FC HC

Pro

po

rtio

n o

f B

ifid

ob

act

eri

um

6hr 12hr

aa

b

x

y

xy NC = No Colostrum

FC = Fresh Colostrum

HC = Heated Colostrum

Heat-treated colostrum increases Bifidobacterium and reduced the colonization of E. coli in the small intestine

(Malmuthuge et al., 2015 )

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3'SLN 6'-SLN 3'-SL 6'-SL DSL

Co

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(µg

/g)

Bovine Colostrum Oligosaccharide

Fresh Colostrum

Heated Colostrum

Heat-treatment may cleave prebiotic oligosaccharides from colostral proteins and lipids

(Fischer et al., 2018)

Heat Treatment of Colostrum

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6SL 6SLN DSL 3SL

Co

nce

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Oligosaccharide

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Oligosaccharides – Transition*

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*

* * *

*

*

*

Bovine colostrum oligosaccharides (bCOs) produced in higher concentrations in milkings 1-3 compared to milking 8+

Multiparous cows have higher concentrations of 6’SLN compared to primiparous (Fischer et al., 2020)

First Feeding Wk 1

Colostrum Solid Feed

Transition

First Feeding

Colostrum Milk

Wk 1

Milk

From Colostrum to Milk

All calves fed one meal of colostrum followed by:

Milk

50% milk/ 50% colostrum (Transition)

Colostrum (Pyo et al., 2020)

Milk 50%/50% Colostrum

From Colostrum to Milk

(Hare et al., in review)

IgG

mg/ml

Hours after birth

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10

20

30

40

0 2 4 6 8 10 12 14 16 18 20 22 24

Second Meal

First Meal

Milk 50%/50% Colostrum

From Colostrum to Milk

Passive Transfer Trancytosis of immunoglobulins

(Jochims et al., 1997)

Receptor mediated and highly regulated

Trancytosis (to blood)

Recycling (back to lumen)

Metabolism (endosome)

Regulation of these pathways in calves is unclear

Endosome Formation

Basal Membrane Release

Recycled to Lumen

Metabolized

Pinocytosis

Industry Concerns

AntimicrobialMaternal

Colostrum Plane of Nutrition

Knowledge Gaps

Normal Pre-Weaning Milk Intake

(de Passille et al., 2016) (Jasper and Weary, 2002)

d4 of life

Feeding Large Meals

Calves typically nurse 6-12 times per day in the first weeks of life (Jensen, 2004)

Larger meals fed less frequently increase the risk of: Abomasal inflammation & lesions

Milk overflow into the rumen

Ruminal acidosis, decreased passage rate and digestion

(Berends et al., 2012; 2015)

Inflamed

Abomasum

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0 60 120 180 240 300 360 420Ace

tam

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en

(m

g/L

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Time (min)

Larger Meal Size and Insulin Sensitivity

Compared calves fed elevated (8L/d) vs low (4L/d) plane of milk 2x per day

No evidence of post-prandial hyperglycemia and hyperinsulinemia

No difference in glucose tolerance

Slower (41% reduction, P = 0.02) abomasal emptying rates during the pre-weaning phase

(MacPherson et al., 2016)

Elevated, KSB = 0.21Low, KSB = 0.34

Gastric emptying rate will influence glucose appearance in blood

(Stahel et al., 2016)

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Glu

cose

(Mm

) an

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lin

(ng

/ml)

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tam

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en

(m

g/m

l)

Time (min)

Glucose

Insulin

Acetaminophen

Gastric Emptying and Glucose-Insulin Dynamics

Glucagon-like Peptides

Proliferation

Nutrient absorption

Gut motility

Blood flow

Gut Permeability

Milk Replacer vs Whole Milk Most MR are high in lactose and osmolarity, low in fat compared with whole milk

0%

20%

40%

60%

80%

100%

Milk MR

other

ash

protein

fat

lactose

18%

45%37%

31%

300 mOsmwhole milk /body fluid

400-600 mOsmMR

Hypertonic MR increases gut permeability (Wilms et al., 2019)

Higher lactose results in increased gastric emptying and lower glucose tolerance in the first week of life (Welboren et al., in review)

Weaning Challenges A smooth transition from a monogastric to a ruminant

Decreases morbidity and mortality and increases gain (Khan et al., 2012)

Requires adequate size and function of the rumen (Baldwin, 2004)

Elevated plane of nutrition pre-weaning makes weaning more challenging (Khan et al., 2011)

Pre and Post-WeaningPre-ruminant Ruminant

Milk Solid Feed

Weaning Transition

1 wk 4 wk 8 wk 12 wk

Abnormal GIT Development Ruminal parakeratosis is

common during weaning(Bush, 1965)

Ruminal acidosis has been documented however to date, no research has linked it to impairment of gut health (Laarman et al., 2012)

Parakeratosis

Is ruminal acidosis good or bad for the calf?

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pH 5.5

pH 5.2

week

6 week wean

Ruminal pH During Weaning

(Kohler et al., in preparation)

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ace

are

a (

µm

2 )

week

6 week wean

Ruminal pH During Weaning

(Kohler et al., in preparation)

Ruminal Gene Expression

3063 557

Common genes are predominately metabolic Ketogenesis

Fatty acid metabolism

It takes time for the genes involved in structural adaptations to change

(Kohler et al., in preparation)

Early and Abrupt Weaning

Pre-ruminant Ruminant

Milk Solid Feed

Transition

Pre-ruminant Ruminant

Milk Solid Feed

Weaning Age

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1 8 15 22 29 36 43 50 57 64

EarlyLate

Calf Age (Eckert et al., 2015)

L/d

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EarlyBW

(kg)

6 week wean

Calf Age (Eckert et al., 2015)

Weaning Age - Bodyweight

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110

0 7 14 21 28 35 42 49 56 63 70

Early

Late

6 week wean

8 week wean

* **

**P<0.05*

Calf Age (Eckert et al., 2015)

Weaning Age - Bodyweight

BW

(kg)

(Eckert et al., 2015)

Weaning Age – ME Intake

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35 42 49 56 63 70

Weaning Strategy – Delayed WeaningImpact on Ruminal Development

Calf Age (d)

Weaning Strategy – Delayed WeaningImpact on Ruminal Development

Wean

6 wk wean

8 wk wean

Wean

Post-weaning

(Meale et al., 2016)

Pre-weaning

L/d

Calf Age (d)

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Step-Down

Abrupt

Step-Down Weaning

(Steele et al., 2017)

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Step-Down

Step-DownWeight (kg)

Weaning

Calf Age (d)

Step-Down

Step-Down - BodyweightP<0.05

(Steele et al., 2017)

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0 6 12 18 24 30 36 42 48 54

AbruptStep-Down

*

Weight (kg)

Calf Age (d)

*

Step-Down - BodyweightP<0.05

(Steele et al., 2017)

WeaningStep-Down

0

2

4

6

8

0 6 12 18 24 30 36 42 48 54

AbruptStep-Down

Intake (Mcal/d)

Calf Age (d)

Weaning

Step-Down

Metabolizable Energy Intake

(Steele et al., 2017)

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2500

30 36 42 48 54

-0.3 -0.2 -0.1 0.0 0.1 0.2 0.3

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-0.2

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PCoA plot_weighted unifrac distance

PC1 Percent variation explained 17.8%

PC

2 P

erce

nt v

aria

tion

expl

aine

d 9.

1% Abrupt_post-weaning

Abrupt_pre-weaning

Step-down_post-weaning

Step-down_pre-weaning

(Meale et al., 2016)

Weaning Strategy – Abrupt WeaningImpact on Ruminal Development

Calf Age (d)

Pre-weaningPost-weaning

Wean

Gradual

Abrupt

Pre and Post-WeaningPre-ruminant Ruminant

Milk Solid Feed

Weaning Transition

1 wk 4 wk 8 wk 12 wk

Fecal microbiota displayed more diversity post-weaning (Meale et al., 2015)

0

2

4

6

8

10

36 48 54

** P = 0.04

Abrupt Weaning – Delayed WeaningImpact on Hindgut

Calf Age (d)

Fecal Starch %

Step-down

Abrupt

Wean

Diversity in Fecal Scores

Barrier Function at Weaning Starter feeding in calves decreased the expression of

tight junctions (Malmuthuge et al., 2012)

Weaned (d 40)

Not Weaned

(Wood et al., 2015)

Barrier Function at Weaning Weaning related changes of the gut epithelium (Pletts et al., in review)

Not-Weaned, d 42

Weaned, d 42

Rumen Duodenum

Endoscopic Biopsy

Post-Weaning and Beyond An area that has not been studied

Need to integrate pre and post weaning planes of nutrition with lifetime performance

Industry Concerns

AntimicrobialMaternal

Colostrum Plane of Nutrition

Knowledge Gaps

Potential Mechanisms of Actions of Saccharomyces cerevisiae boulardii

(SCB)

1st line of defense:Intestinal microflora

3rd line of defense:Intestinal immune system

2nd line of defense:Intestinal epithelia

Anti-inflammatory

properties, immune cells

recruitment, cytokine

production, etc...

Intestinal permeability, mucin

production, antimicrobial peptides,

etc...

Reinforcement of

positive microflora : LAB,

cellulolytic bacteria…

42 calves from d2 to d54

2 groups: CON and SCB fed MR + starter ad libitum

No difference on performance and health between CON and SCB

BUT: overall health = very good

adequate passive transfer, clean environment, no transportation

Looking at more challenging conditions closer to on-farm reality

Study 1: Pre-weaning

(He et al., 2017)

Study 2: Veal farm 2 groups: CON & SCB fed MR only

Back to basics: verification of the successful supplementation of SCB

Copy number of 26S rRNA Saccharomyces cerevisiae gene/g of feces

0.0

0.5

1.0

1.5

2.0

2.5

0 2 7

CON

SCB

TRT = 0.032

Week = 0.001

TRT*Week = 0.008

10

7copy n

um

ber/

g

Week

Reduction in fecal score and total cases of diarrhea

Study 2: Veal farm challenge

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1 2 3 4 5 6 7

Week of experiment

*

*

Total # of diarrhea cases

0.6

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Week of experiment

*

**

Fecal score

* P < 0.05* P < 0.05

(Villot et al., 2019)

Study 2: Veal farm challenge No big changes highlighted in microbiota composition of feces

Difficult to understand the mechanisms of action with on-farm experiment

(Villot et al., 2019)

Initial health status

Transportation

Housing conditions

Antibiotic treatment

New strategy:

Start treatment at birth

Supplementation from colostrum feeding in controlled conditions

Investigation of the gut with dissection

Looking at microbiota + host response

Confounding factors?

Study 3: Neonate

Dr. Clothilde Villot

Endoscopic biopsy

Study 3: Neonate

d0 d1 d2 d3 d4 d5 d6 d7

Remove from the dam

directly after birth

h2 h12 19:00

B

I

R

T

H

Milk Replacer 17% of BW

+ SCB/Control

7:00 07:00

…

E

U

T

H

A

N

A

S

I

A

Fecal sample & score & body temperature

Dissection

Passage of nutrients

7:00

10:00

7:00

Blood sample

Milk Replacer 15% of

BW

+ SCB/Control

Colostrum

+

SCB/Control

GUT Permeability

Study 3: Neonate Colon biopsy technique:

Non invasive Repeated measurements Good quality of samples

Measurements: Bacterial composition with qPCR & DNA sequencing (16S amplicon) Genes expression of the host with total RNA sequencing

(Van Niekerket al., 2018)

H&E stain

Recovery of Cr in urine, %

CON (mean ± SE)

SCB(mean ± SE)

P-Value (Student Test)

Day TRT TRT * Day

Day 2 3.01 ± 0.12 3.44 ± 0.15 NS 0.863 NS

Day 6 3.40 ± 0.19 2.53 ± 0.08 0.080

Gut permeability test

No difference of gut permeability in d2 and d6

Gut permeability tends to be improved at d6 for SCB calves

Study 3: Neonate

Study 3: Neonate

Provides mucosal immune protection in a non-inflammatory manner

Promotes the maintenance of appropriate bacterial communities

Neutralization of incoming pathogens and microbial inflammatory products, and facilitates antigen signaling

(Mutwiri et al., 1999)

Immunoglobulin A

Immunoglobulin A Production

0

0.5

1

1.5

2

2.5

Jejunum Ileum Colon Feces

IgA mg/g Dry Matter

**

Endogenous production of IgA in the gut of 7d old calves IgA production in ileum SCB seems to stimulate IgA production

* P < 0.05

(Villot et al., in review)

Immunoglobulin A Production

Endogenous production of IgA in the gut of 7d old calves IgA production in ileum SCB seems to stimulate IgA production

(Villot et al., in review)

Take Home Messages There are still some basic concepts in calf biology and

nutrition that we do not understand

No difference between tube vs. bottle feeding colostrum for passive transfer

Delaying colostrum by six hours can impact passive transfer and gut microbiology

Pasteurizing colostrum may help to improve calf gut health if managed properly

Take Home Messages An abrupt transition from colostrum to milk can

compromise gut development

Elevated planes of milk can be fed early in life

Elevated planes of milk can be fed with 2x/day feeding schemes

Milk replacer formulations high in lactose may impact gut health and insulin sensitivity

Take Home Messages Weaning in dairy calves is one of the largest

transformations of the gut in nature

Weaning age and abruptness impact performance on high planes of milk nutrition

Weaning is also associated with gut health problems

Post-weaning nutrition is another area left undiscovered in calf nutrition

Direct fed microbials have potential to improve gut health in pre-weaned calves