Amino Acid Balancing for the TransitionCow: Old and New Stories from a

Molecular Perspective

Johan Osorio, Assistant Professor

Dairy and Food Science Department

South Dakota State University, Brookings, USA

Amino acid balancing for the transition cow: Old and

new stories from a molecular perspective

June 10, 2020

Johan OsorioAssistant ProfessorDairy and Food Science DepartmentSouth Dakota State University, Brookings, USA

(Block et al., 2001)

Energy balance

Body condition score

Energy intake

4% Fat corrected milk

(Ingvartsen, 2006)

Peripartal or Transition periodTissue mobilization

First lactationThird lactation

Pre calving Post calving

~3 4 wk ~3 4 wkCalving

Cow in negative energy balance

NEFA

TG

NEEpi Insulin

NEFA

NEFA

CO2

Propionate

Ketonebodies

Mitochondria

Glucose

AA, Lactate andGlycerol

TG

FeedIntake

TG

VLDL

Milk Fat

Modified from Drackley, 1999

Decrease

Increase

Body fat and protein mobilization

9574 74

9574 74

143

97 83

186

125103

0

50

100

150

200

250

300

2 5 12 2 5 12

Body

fata

ndprotein,

kg

Week relative to parturition

FatProtein

16% Crude protein 19% Crude protein

32.2%32.8%

22.1% 22.1%

(Komaragiri and Erdman, 1997)

Peripartal protein mobilization

97.2

88.9 89.5

84

86

88

90

92

94

96

98

14 60 120Day relative to parturition

Body protein (kg)

10.4

17.8

8.46.9 6.7 7.6

6.2 5.4

02468

101214161820

7 7 14 28 56 84 112 119Day relative to parturition

3 Methylhistidine (μM)

(Adapted from Phillips et al., 2003; J Dairy Sci 86:3634 47)

Proteolysis 3 Methylhistidine

Metabolizable protein and amino acids likely limiting around calving

Bell et al. (2000)

Metabolizable Protein

Needed for milk protein, glucose synthesis, synthesis of other compounds (e.g. SAM, glutathione, taurine)

1 2

3 4

1

5 6

(Larsen and Kristensen, 2013)

Net liver uptake of Methionine and Histidine increases after calving

Net uptake of Met by liver can be enhanced by supplemental RP-Met……..also prevents decrease in blood Met postpartum (Dalbach et al., 2011)

Methionine and the Peripartal Period

Dietarycomponent

Osorio et al., 2013 Zhou et al., 2016 Batistel et al., 2017

Control Met Control Met Control Met

CP, % of DM 17.4 17.4 17.2 17.3 17.7 17.7

MP supplied (g/d) 1,563 1,840 2,090 2,374 2,425 2,640

MP balance (g/d) 574 616 434 573 118 160

Lys (% of MP) 6.17 6.07 6.33 6.24 6.40 6.38

Met (% of MP) 1.81 2.15 1.79 2.30 1.70 2.24

Lys:Met 3.43:1 2.82:1 3.54:1 2.71:1 3.78:1 2.88:1

Methionine and the Peripartal Period

Day relative to parturition

0 5 10 15 20 25 30 35

Milk

yie

ld (k

g/d)

0

10

20

30

40

50

ControlMethionine

Adapted from Osorio et al., 2013 Adapted from Zhou et al., 2016

Adapted from Batistel et al., 2017

+ 3.4 kg + 3.8 kg

+ 4.2 kg

Milk Yield

Methionine

Milk yield & Performance

Antioxidant

Liver function

DNA &Histonemethylation

Reduced immunosuppression

Liver Function

Day relative to parturition

0 5 10 15 20 25 30 35

Milk

yie

ld (k

g/d)

15

20

25

30

35

40

45

50

55

Mild (< 5% TG)Moderate (5-10% TG)Severe (> 10%)

FL, P = 0.25DIM, P < 0.01FL X DIM, P < 0.01

Fatty liver on milk yieldn = 100 transition cows

Actual TG mean:• Mild = 3.3 ± 1.0• Moderate = 6.5 ± 1.5• Severe = 11.1 ± 0.9

Actual Milk yield:• Mild = 41.9 ± 0.84• Moderate = 41.6 ± 1.5• Severe = 36.9 ± 2.9

Correlation• r = 0.2• P = 0.06

Osorio et al., 2013; Zhou et al., 2016; Bastistel et al., 2017

FL = Fatty liver level

7 8

9

11 12

10

2

Methionine plays several roles in liver

TG

VLDL, secreted from liver

TG

Phospholipid synthesis

Choline + FA +

+

Acetate

Apolipoprotein synthesis

Amino acids (Met, et al.)

+

NEFA

Esterification

Met

Peripartum

GlutathioneTaurine(antioxidants)

Albumin Major secretory protein synthesized by liver

Liver functionCarnitine metabolism

Liver functionCarnitine metabolism

Day relative to calving

777777 21212121777 212177 217 217 217 21

Free

car

nitin

e (n

mol

es/g

of w

et ti

ssue

)

0

20

40

60

80

100

120

140

160

180 CONMSSM

Free carnitine Met P < 0.01

Osorio et. al., 2014

ApoB

100

, ug/

mL

460

480

500

520

540

Control Methionine

ApoB

100

(ng/

mL)

0

5

10

15

20

25

30

Control Methionine

Liver function

Diet, P = 0.16Time, P = 0.97D × T, P = 0.75Contrast, P = 0.14

Adapted from Osorio et. al., 2013

VLDL = Very low density lipoproteins

Adapted from Sun et. al., 2016

VLD

L, m

mol

/L

0.44

0.46

0.48

0.50

0.52

0.54

0.56

0.58

Control Methionine

Adapted from Sun et. al., 2016

ApoB-100

ApoB-100VLDL

Diet, P = 0.001 Diet, P = 0.008

Inflammatory response

Local acuteInflammatory response

+ APP:Serum amyloid A (SAA)HaptoglobinCeruloplasmin

Kuby Immunology, 2007

Metritis

Mastitis - APP:AlbuminApolipoproteins

Liver

Inflammatory response

Metritis Mastitis

Liver

Potential Pathogen Infection

AcutePhaseProteins

13 14

3

15 16

17 18

Fat or Adipose Tissue

NEFA

TG

NEFA

Liver

InsulinNE, Epi

(modified from Drackley, 1999)

Cow in negativeenergy balance

InflammatoryResponseSignal

NEFA

Inflammatory response

Metritis Mastitis

Liver

Potential Pathogen Infection

AcutePhaseProteins

Albumin

Liver function

Day relative to parturition-21-21-21-21 -10-10-10 77 14-21-21-21-21-21-21-21-21-21-21-21-21-21-21 -10-10-10-10-10-10-10-10-10-10-10-10-10 777777777777 1414141414141414141414 21212121212121212121-21-21-21-21-21-21-21-21-21 -10-10-10-10-10-10-10-10 7777777 141414141414 2121212121-21-21-21-21 -10-10-10 77 14-21 -10 7 14 21-21 -10 7 14 21-21 -10 7 14 21

Alb

umin

(g/L

)

33

34

35

36

37

38

CONMSSM

Bertoni et. al., 2008. J Dairy Sci

Diet, P = 0.28Time, P = 0.04D × T, P = 0.20Contrast, P = 0.15

Osorio et. al., 2014

Milk yield +6.5 kg/d in Upper

MS = MetasmartSM = Smartamine M

Dietary methyl-donors in dairy cows

SAM

SAHHomocysteine

Cysteine

Diet TissuesMilkColostrum

Liver Function

DNA methylation Epigenetics

VLDLPC

Vit. B12 CH3

PE

rRNA complex Protein synthesis initiation

Choline

Betaine

Dimethylglycine

Diet

5-MTHF

Methionine

Diet

Folate+ + +

Cooke et al. 2007Zom et al. 2011

+

GlutathioneAntioxidants

Taurine+

EmbryoFetus

Day relative to parturition

777777 21212121777 212177-10 7 217 217 21

Glu

tath

ione

(mM

)

0.0

0.5

1.0

1.5

2.0

2.5

3.0

3.5

CONMSSM

Glutathione Met P = 0.04

Liver functionGlutathione Priming Effect

Osorio et. al., 2014

Batistel et al., 2018

Zhou et al., 2016

-10.0 7.0 21.0

1

0.2

0.3

0.4

0.5

0.6

0.7

0.8

0.9

2

3

4

5

-10.0 7.0 21.0

1

0.2

0.3

0.4

0.5

0.6

0.7

0.8

0.9

2

3

4

5

-10.0 7.0 21.0-10.0 7.0 21.0

MethionineControl

Methionine supplementation alters the liver transcriptome2,663 genes with diet time effect

Gene expression

10 7 21 10 7 21Day relative to parturitionOsorio et al., 2012

19 20

21 22

23 24

4

Methionine and Gene RegulationCentral Dogma of Molecular Biology

Epigenetic Mechanisms

Methionine and Gene RegulationHistone

Inactive/closeInactive/close

Active/OpenActive/Open

Methionine and NutrigenomicsHistone Methylation

Tiramisu

Histone Methylation

0

0.1

0.2

0.3

0.4

0.5

0.6

0.7

0.8

0.9

0 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30

Relative Histone Methylation

Control 125 uM 250 uM 500 uM

Control

125 uM Met

250 uM Met

500 uM MetFernanda Rosa

Histone MethylationMethionine cycle

Enzymes

Fernanda Rosa

Histone MethylationMethionine cycle

Enzymes

Fernanda Rosa

25 26

27 28

29 30

5

Histone Methylation

Total protein

Milk protein

0

0.2

0.4

0.6

0.8

1

0 2 4 6 8 10 12 14 16 18 20 22 24 26 28 30

Relative Histone Methylation

Control 125 uM 250 uM 500 uM

Fernanda Rosa

Histone Methylation

DietaryMethionine?

DietaryMethionine?

Bionaz et al., 2012; PLoS ONE 7:333268Day relative to parturition

Euchromatin

Heterochromatin

HistoneMethylation

ConclusionsMethionine

Performance

PostpartumPostpartumPrepartum

Gene Regulation

Performance

Metabolism

Gene expression

MethionineModel

?

31 32

33 34

6

7

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