Amino acid metabolism in periparturient dairy cattle

H. Lapierre, D.R. Ouellet, M. Larsen and L. Doepel

Agriculture and Agri-Food CanadaAarhus University – Foulum, Denmark

Trouw Nutrition, Canada

International Dairy Nutrition Symposium

October 2017

Fairly little focus despite acknowledged

Postpartum protein deficiency

(Bell et al. 2000)MP:

meta

bol

izab

le p

rote

in

1. AA metabolism post-calving2. From pre- to post-calving3. Can we reduce the deficit?

AA supplementation4. Other roles of AA

glutamine supplementation5. Conclusions

AA metabolism

liver

hepatic

LIVER

artery

portal PDV

AA across tissues

POST-LIVER

=SPLANCHNIC

MPY

10 cows prepared with splanchnic catheters before parturition

4 Control (post-rumen water infusion)Blood samples collected on DIM 5, 15

and 29

1. AA metabolism post-calving

Postpartum protein deficiency

(Larsen et al. 2015; NRC 2001)

(Larsen et al. 2015; NRC 2001)

Postpartum protein deficiency

-571 -291

-64

Lysine net flux (mmol/h)

(Larsen et al. 2015)

(Larsen et al. 2015)

Lysine net flux (mmol/h)

(Larsen et al. 2015)

Lysine net flux (mmol/h)

(Larsen et al. 2015)

Group 2 AAIle, Leu, Val

Lysine net flux (mmol/h)

(Larsen et al. 2015)

Methionine net flux (mmol/h)

(Larsen et al. 2015)

Methionine net flux (mmol/h)

(Larsen et al. 2015)

Methionine net flux (mmol/h)

(Larsen et al., 2015)

Group 1 AA: His, Phe+Tyr, Trp

Methionine net flux (mmol/h)

(Larsen et al., 2015)

Methionine net flux (mmol/h)

Uptake:outputU:O

(Larsen et al. 2014 & 2015)

Mammary uptake : output

?

Leucine: 13C from 13C-Leucine was recovered in 13CO2 ->oxidation = energy (Raggio et al. 2006)

Lysine: 15N from 15N-Lysine was recovered into milk non EAA (Asx, Glx,Ser and Ala;Lapierre et al. 2008)

EAA: in vitro, labelled AA into milk lactose (Bequette et al. 2006)

Excess uptake is used for what?

(Larsen et al. 2015)

NEAA net flux (mmol/h)

(Larsen et al. 2015)

NEAA net flux (mmol/h)

(Larsen et al. 2015)

NEAA net flux (mmol/h)

DIM 5: post-liver supply of AA clearly

insufficient to cover MPY utilization of body proteinsDIM 29: post-liver supply of AA ≤ MPY

« Pattern » similar to establishedlactation for Groups 1 and 2-AA

1. AA metabolism post-calving

6 dairy cows

Pre-calving:18 days before calving1326 g MP/d

Post-calving: 21 or 42 DIM2136 g MP/d40.2 kg/d milk

2. Pre- vs. post-calving

(Doepel et al. 2009)

Lysine net flux (mmol/h)

(Doepel et al. 2009)

Lysine net flux (mmol/h)

(Doepel et al. 2009)

Lysine net flux (mmol/h)

(Doepel et al. 2009)

Methionine net flux (mmol/h)

(Raggio et al. 2004)

Methionine net flux (mmol/h)

Established lactation

(Doepel et al. 2009)

Methionine net flux (mmol/h)

(Doepel et al. 2009)

Methionine net flux (mmol/h)

(Doepel et al. 2009)

Methionine net flux (mmol/h)

(Doepel et al. 2009)

Methionine net flux (mmol/h)

Liver/Portal 0.62

Liver/Portal 0.32

liver

Liver inflow of AA

+ PDV

arterial input

liver

Liver inflow of AA pre-calving

PDV + arterial inflow

liver

PDV + arterial inflow

Liver inflow of AA post-calving

(Doepel et al. 2009)

Methionine net flux (mmol/h)

Liver/Inflow 0.11

Liver/Inflow 0.08

(Doepel et al. 2009)

Non-essential AA net flux (mmol/h)

(Doepel et al. 2009)

Non-essential AA net flux (mmol/h)

The comparison helped to delineatethat the liver is not « THE » key controlbut acts in response to both the supplyand utilization of AA by other tissues

At the initiation of lactation, the liver« spares » AA: no increment of AAremoval AA to support gluconeogenesis

2. Pre- vs. post-calving

3. Can we reduce the deficit?

2 studies with post-rumen infusion1 field study

2 studies with post-rumen infusion:

« Close-the-gap » strategy Study 1: casein (CN: 720 to 194 g/d) Study 2: free AA, CN profile (791 to 226 g/d)

Days relative to parturition

4 15 294 15 294 15 294 15 294 15 294 15 294 15 294 15 29

Net

rele

as

e -

milk s

ec

reti

on

, g

/d

-300

-200

-100

0

100

200

300 PDV release

TSP release

n = 18Larsen & Kristensen, 2012Raun & Kristensen, 2011Dalbach et al., 2011Larsen & Kristensen, 2009a,b

Days relative to calving

0 5 10 15 20 25 30

Am

ino

acid

s, g

/d

0

100

200

300

400

500

600

700

800

900

Sampling

Sampling

Sampling

(Larsen et al. 2014)

Milk protein yield (MPY) in response to CN infusion (study 1)

Efficiency > 70%

Days relative to calving

-15 -10 -5 0 5 10 15 20 25 30

kg

/d

0

10

20

30

40

50

60 Ptrt < 0.01, PDIM < 0.01, Ptrt x DIM = 0.29

46.0 ± 0.8

38.2 ± 0.9

Milk yield in response to AA-CN infusion (study 2)

• 7.8 ± 1.3 kg greater with AA-CN• Similar to +7.2 kg infusing CN

Efficiency = 45%

(Larsen et al. 2015)

MP balance in response to AA-CN infusion (study 2)

(Larsen et al. 2015)

MP balance in response to AA-CN infusion (study 2)

Ptrt = 0.30

(Larsen et al. 2015)

Group 1-AA net flux (mmol/h)

His, Met, Phe+Tyr, Trp

(Larsen et al. 2015)

Group 1-AA net flux (mmol/h)

95% recovery

*

(Larsen et al. 2015)

Group 1-AA net flux (mmol/h)

*

*

**

(Larsen et al. 2015)

Group 2-AA net flux (mmol/h)

Ile, Leu, Lys,Val

(Larsen et al., 2015)

Group 2-AA net flux (mmol/h)

*

(Larsen et al., 2015)

Group 2-AA net flux (mmol/h)

**

*

*

(Larsen et al., 2015)

Group 2-AA net flux (mmol/h)

**

*

(Larsen et al. 2015)

NEAA net flux (mmol/h)

(Larsen et al. 2015)

NEAA net flux (mmol/h)

*†

†

?

(Galindo et al. 2015)

Liver glucose total flux (mmol/h)

=

(Galindo et al. 2015)

Glucose total flux (mmol/h)

†

(Galindo et al. 2011)

Glucose total flux (mmol/h)

*

Established lactation

NEL balance in response to AA-CN infusion (study 2)

Ptrt×DIM = 0.10

(Galindo et al. 2015)

*

†

(Galindo et al. 2015)

[NEFA], mM

-> increased fat mobilisation at DIM 5

*Ptrt×DIM = 0.05

(Galindo et al. 2015)

[BHBA], mM

*† Ptrt×DIM = 0.03

Field trial (preliminary results): 91 Holsteins randomised block design

PMR 3 kg conc./d

PMR with extra protein 3 kg conc./d

PMR 3 kg conc. + 2 kg barley/d

Control~15.5% CP

Protein~20.5% CP

Energy~15.0% CP

Calv. 14 d 29 d

Same feed

Wk 1 to 4: +5.5 kg/d for older cows

Primi Multi

kg

/d

0

10

20

30

40

50

CONTROL

PROTEIN

ENERGY

Pdiet x parity < 0.01

Milk yield

(Larsen et al. EAAP, 2017)

Milk protein and fat yield followed milk yield

• Concentrations did not differ among diets72

Milk fat yield

Primi Multi

kg

/d

0.0

0.5

1.0

1.5

2.0

CONTROL

PROTEIN

ENERGY

Pdiet x parity < 0.05

Milk protein yield

Primi Multi

kg

/d

0.0

0.5

1.0

1.5

2.0

Pdiet x parity < 0.05

(Larsen et al. EAAP, 2017)

Greater fat mobilisation with high protein allocation

Plasma NEFA

Week after calving

1 2 3 4 5 81 2 3 4 5 81 2 3 4 5 81 2 3 4 5 81 2 3 4 5 81 2 3 4 5 8

M

0

200

400

600

800

1000

C multi

P multi

E multi

Pdiet x parity x week

< 0.01

(Larsen et al. EAAP, 2017)

BHBA tended to be greatest with control

74

Plasma BHB

Week after calving

1 2 3 4 5 81 2 3 4 5 81 2 3 4 5 81 2 3 4 5 81 2 3 4 5 81 2 3 4 5 8

mM

0.2

0.4

0.6

0.8

1.0

1.2

1.4

C multi

P multi

E multi

Pdiet

= 0.07; Pparity

< 0.01

(Larsen et al. EAAP, 2017)

3. Can we reduce the deficit?

Increased protein supply increased MPY -> failed to decrease protein deficiency!

BUT: increased AA concentrations!!!

Although the deficit was not reduced:

CN increased fractional synthesisrate of albumin at DIM 4

CN increased rumen papillaeproliferation

CN stabilized inflammatoryresponsiveness of leukocytes

(Larsen et al. 2017)

4. Other effects of AA

Glutamine: conditionally essential NEAAImmune system (lymphocyte, cytokine)Precursor or purine and pyrimidine synthesisMajor energy sourceGlu + Gln ≈ 20 % AA in milkPlasma concentrations still low at 29 DIM-> post-rumen infusion of 300 g/d Gln –

21 days post-calving(Doepel et al. 2006)

20

24

28

32

36

40

44

4 7 11 14 18 21

Day from calving

Mil

k,

kg

/d

Ctl Gln

Effect of Gln infusion

non significant increment of milk yield

83% recovery of infused Gln in the portal vein

(Doepel et al. 2006 & 2007)

no effect on immune parameters

5. Conclusions: AA metabolism

AA deficiency mirrors estimated MP balance: post-liver vs. mammary uptakeAA metabolism post-calving follows the same pattern as in established lactation: Group 1:

• liver catabolism• Mammary U:O of EAA ≈ 1

Group 2: • Very little liver catabolism• Mammary U:O of EAA > 1; but decreases with low

supply

5. Conclusions: pre- vs. post-calving

The liver is NOT the key regulator Responds to both absorption and tissue

utilization

AA priority is to make protein Initiation of lactation does NOT

increase AA liver removal

5. Conclusions: reducing the deficit

Increased AA supply: Very efficient use of extra AA into MPY Does not reduce AA deficiency No increment of liver gluconeogenesis Increased energy deficiency But limited effect on [NEFA & BHBA] Protein appears more limiting than energyin the month post-calving Beneficial to reach the biological

potential of a larger production?

Thanks to:Dairy Farmers of Canada,Dairy Farmers of Québec (Novalait)Aarhus University, DenmarkAgriculture and Agri-Food CanadaAjinomoto Heartland, Inc.Danish Agri-Fish AgencyDanish Council for Independent ResearchDanish Council for Technology and InnovationDanish Milk Levy FundEvonik Industries AGMinistry of Food, Agriculture and Fisheries,

DenmarkNatural Science and Engineering

Research Council of Canada (NSERC)

Questions?

Thank you!