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Amino acid metabolism in periparturient dairy cattle
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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
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Fairly little focus despite acknowledged
Postpartum protein deficiency
(Bell et al. 2000)MP:
meta
bol
izab
le p
rote
in
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1. AA metabolism post-calving2. From pre- to post-calving3. Can we reduce the deficit?
AA supplementation4. Other roles of AA
glutamine supplementation5. Conclusions
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AA metabolism
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liver
hepatic
LIVER
artery
portal PDV
AA across tissues
POST-LIVER
=SPLANCHNIC
MPY
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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
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Postpartum protein deficiency
(Larsen et al. 2015; NRC 2001)
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(Larsen et al. 2015; NRC 2001)
Postpartum protein deficiency
-571 -291
-64
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Lysine net flux (mmol/h)
(Larsen et al. 2015)
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(Larsen et al. 2015)
Lysine net flux (mmol/h)
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(Larsen et al. 2015)
Lysine net flux (mmol/h)
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(Larsen et al. 2015)
Group 2 AAIle, Leu, Val
Lysine net flux (mmol/h)
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(Larsen et al. 2015)
Methionine net flux (mmol/h)
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(Larsen et al. 2015)
Methionine net flux (mmol/h)
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(Larsen et al. 2015)
Methionine net flux (mmol/h)
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(Larsen et al., 2015)
Group 1 AA: His, Phe+Tyr, Trp
Methionine net flux (mmol/h)
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(Larsen et al., 2015)
Methionine net flux (mmol/h)
Uptake:outputU:O
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(Larsen et al. 2014 & 2015)
Mammary uptake : output
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?
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?
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(Larsen et al. 2015)
NEAA net flux (mmol/h)
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(Larsen et al. 2015)
NEAA net flux (mmol/h)
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(Larsen et al. 2015)
NEAA net flux (mmol/h)
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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
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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
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(Doepel et al. 2009)
Lysine net flux (mmol/h)
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(Doepel et al. 2009)
Lysine net flux (mmol/h)
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(Doepel et al. 2009)
Lysine net flux (mmol/h)
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(Doepel et al. 2009)
Methionine net flux (mmol/h)
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(Raggio et al. 2004)
Methionine net flux (mmol/h)
Established lactation
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(Doepel et al. 2009)
Methionine net flux (mmol/h)
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(Doepel et al. 2009)
Methionine net flux (mmol/h)
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(Doepel et al. 2009)
Methionine net flux (mmol/h)
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(Doepel et al. 2009)
Methionine net flux (mmol/h)
Liver/Portal 0.62
Liver/Portal 0.32
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liver
Liver inflow of AA
+ PDV
arterial input
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liver
Liver inflow of AA pre-calving
PDV + arterial inflow
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liver
PDV + arterial inflow
Liver inflow of AA post-calving
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(Doepel et al. 2009)
Methionine net flux (mmol/h)
Liver/Inflow 0.11
Liver/Inflow 0.08
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(Doepel et al. 2009)
Non-essential AA net flux (mmol/h)
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(Doepel et al. 2009)
Non-essential AA net flux (mmol/h)
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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
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3. Can we reduce the deficit?
2 studies with post-rumen infusion1 field study
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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
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(Larsen et al. 2014)
Milk protein yield (MPY) in response to CN infusion (study 1)
Efficiency > 70%
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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%
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(Larsen et al. 2015)
MP balance in response to AA-CN infusion (study 2)
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(Larsen et al. 2015)
MP balance in response to AA-CN infusion (study 2)
Ptrt = 0.30
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(Larsen et al. 2015)
Group 1-AA net flux (mmol/h)
His, Met, Phe+Tyr, Trp
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(Larsen et al. 2015)
Group 1-AA net flux (mmol/h)
95% recovery
*
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(Larsen et al. 2015)
Group 1-AA net flux (mmol/h)
*
*
**
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(Larsen et al. 2015)
Group 2-AA net flux (mmol/h)
Ile, Leu, Lys,Val
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(Larsen et al., 2015)
Group 2-AA net flux (mmol/h)
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*
(Larsen et al., 2015)
Group 2-AA net flux (mmol/h)
**
*
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*
(Larsen et al., 2015)
Group 2-AA net flux (mmol/h)
**
*
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(Larsen et al. 2015)
NEAA net flux (mmol/h)
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(Larsen et al. 2015)
NEAA net flux (mmol/h)
*†
†
?
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(Galindo et al. 2015)
Liver glucose total flux (mmol/h)
=
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(Galindo et al. 2015)
Glucose total flux (mmol/h)
†
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(Galindo et al. 2011)
Glucose total flux (mmol/h)
*
Established lactation
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NEL balance in response to AA-CN infusion (study 2)
Ptrt×DIM = 0.10
(Galindo et al. 2015)
*
†
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(Galindo et al. 2015)
[NEFA], mM
-> increased fat mobilisation at DIM 5
*Ptrt×DIM = 0.05
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(Galindo et al. 2015)
[BHBA], mM
*† Ptrt×DIM = 0.03
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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
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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)
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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)
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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)
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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)
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3. Can we reduce the deficit?
Increased protein supply increased MPY -> failed to decrease protein deficiency!
BUT: increased AA concentrations!!!
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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)
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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)
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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
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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
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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
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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?
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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)
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Questions?
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Thank you!