Lysine as an amino acid in ruminants: where are we ? Matthieu Rolland; Ajinomoto Animal Nutrition Europe
Lysine as an amino acid in ruminants: where are we ?Matthieu Rolland; Ajinomoto Animal Nutrition Europe
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Why should we care about lysine ?
Sources of Metabolisable Lysine, feedstuffs
& microbial protein
Lysine requirements and conversion
efficiency (components)
Dairy cow response to lysine – meta
analysis (volume)
Guidelines for balancing ration during
lactation
Lysine Functions during Transition
Period, Recommandation for close-up
Presentationoutlines
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Lysine, Essential (& often limiting) Amino Acid for Milk Production
Lys (% CP)
Milk 7.6
Forages 2.8-4.7
Grains 2.8-3.6
Plant Proteins 2.8-6.1
Fiber By-products 4.0-6.3
Lysine infusion resulted in 16% of the total response in yield of milk
protein that was obtained with either the 10 essential amino acids or
sodium caseinate.
Lysine & methionine together accounted for 43% of the total
response.
This suggested that lysine & methionine were first and second
limiting or co-limiting for secretion of milk protein.
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For nutritionists, lysine supply is a better predictor of milk protein yieldpotential than CP or MP
6
Metabolizable Protein provides
a better prediction
Schwab, C. G. (2014). Getting comfortable with amino acid balancing. Penn State Dairy Cattle Nutrition Workshop.
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Metabolizable Lysine : Rumen Microbes & By-pass feedstuffs do the pair
8
% Lysine in feedstuff Rumen Undegradable Protein
Microbial Protein, most valuable source of lysine
Mammary gland uses Amino Acids from the Metabolizable Protein to
synthetize milk protein.
2 main sources of Metabolizable Lysine :
1- Maximize Rumen contribution
2- Select carefully by-pass ingredients
Optimal
MP-Lysine
supply
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Maximizing Microbial Protein & MP-Lysine with high NFC ration
9
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Carbohydrates have greater impact than protein
10
Impact of
CP
variation
Impact of NFC
variation
Broderick, 2003
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11
Predicted ration supply
= 2500 g MP
MP origin = 60%
Microbial, 40% RUP
Actual Rumen MP = 1275 g
Actual Lysine from Microbial Protein = 100.7 g
Assume CHO digestibility overestimate rumen contribution by 15%
Actual ration supply
= 2 275 g MP
Lysine supply
= 152 g
Lysine profile =
6.7 %MP
Lysine profile
= 6.8 %MP
Total Lysine supply = 170 g
Lysine from Microbial
= 118.5 g
170 g MP-Lys = 1206 g milk protein
152 g MP-Lys = 1083 g milk protein
37.7 kg milk at 3.20 % protein
34.1 kg milk at 3.18 % protein
Rumen Carbohydrates impact AA status of dairy cows
Accurate evaluation of forages & grains digestibility is
critical to optimize MP-Lysine supply and profile
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Since mid-70’s models provided guidelines for lysine focusing on milk %protein
NRCINRA
Rulquin (INRA) & Schwab (NRC) established ideal MP-Lys at 7.4 & 7.2 %MP
Guidelines for field application MP-Lys 6.8 & 6.6 %MP
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1996, a first review of production response to lysineAMINO ACIDS AND THEIR APPLICATION IN
FORMULATING DIETS FOR CATTLE
Charles G. Schwab Department of Animal and
Nutritional Sciences University of New Hampshire
Production responses include variable increases in content and yield of milk protein, milk production, and feed intake.
Florida Ruminant Nutrition Symposium. 1996
First, the sequence of Lys and Met limitation is determined by their relative concentrations in RUP.
Second, content of milk protein is more responsive than milk yield to supplemental Lys and Met, particularly in post-
peak lactation cows. In regard to milk protein content, it is noteworthy that responses occur within the first couple of days,
that responses remain similar or become greater after peak lactation, that responses are independent of level of milk yield
and that casein is the milk protein fraction that is most affected and not the whey or NPN fractions.
Third, milk protein responses generally are greater when Lys and Met are supplied together rather than when either
AA is supplied alone.
Fourth, milk protein responses to Lys plus Met are greater when basal levels of either CP or RUP are low rather than high.
Fifth, increasing duodenal concentrations of Lys and Met increases content of milk protein more than would be
expected by increasing ration CP.
And sixth, milk yield responses to Lys and Met are limited generally to cows in early lactation when the need for
absorbable AA, relative to absorbable energy, is the highest.
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METABOLIZABLE PROTEIN AND AMINO ACID NUTRIT ION OF THE COW: WHERE ARE WE IN 2007 C . G . Schwab and S . E . Boucher Depar tment o f An ima l and Nut r i t iona l Sc iences Un ivers i t y o f New Hampsh i re and B . K . S loan Adis seo USA, Inc .
Schwab et al. (2004) showed the overall efficiency of utilization of MP for milk protein secretion to be only of the
order of 0.64 compared to the NRC book value of 0.67,
whereas MP utilization was calculated to be greater than 0.67 when balancing for Lys and Met was integrated
into the formulation approach.
• Milk protein secretion increased in a linear fashion. The optimum response was an extra 173 g of milk
protein (2.7 kg milk, +0.2% milk protein) to increasing daily MP-Lys up to an addition of 34 g.
• The efficiency of utilization of MP for milk protein synthesis was only 0.53 for the imbalanced ration
without any supplemental Lys supplementation.
• At the optimum level of Lys supplementation, the efficiency of utilization of MP was improved to
0.67.
Piepenbrink et al. (1999) fed a Met enriched ration, and studied in a dose response manner
using a replicated Latin square design, the response to increasing supplies of Lysine
For maximal efficiency, lysine needs to be balanced with méthionine
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Balancing ration for MP-Lysine : Grams or %MP ?
Grams of MP-Lysine will determine the potential of milk protein yield
Lysine profile in MP will influence the efficiency of utilisation
Define the supply in grams of MP-Lysine for
a target milk production level
Optimize MP, Lysine %MP for efficient conversion in
milk protein
The Latest on Amino Acid Feeding – 2003 Charles G. Schwab, Ryan S. Ordway, and Nancy L. Whitehouse Department of Animal and Nutritional Sciences University of New Hampshire
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Predicting cow milk volume response to Lysine Meta analysis & modelling by Dr.Izuru Shinzato
Studies controlled by Universities / Institutes,
Studies published in peer-reviewed journals or
presented in ADSA National Meeting
RP Amino Acid added “on-top” of the control diet, no
others variations.
Bioavailability of the RP products used was confirmed
prior to the study
Only multiparous cow data. Most of studies were using
early to peak lactation cows
Energy unlikely limiting (NEl of basal diet no less than
0.78 Mcal/lb DM)
CPM Dairy calculations with analyzed values for
ingredients used in diets
List of studies with RP-Lysine (± RP-Methionine) included in the meta analysis
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Milk volume response to lysineMilk kg response to MP-Lys g/d (supply)
Milk kg response to MP-Lys %MP (profile)
• Clear trend to increase milk volume when
– supply in grams is increased
– profile Lys %MP is improved
• BUT initial milk level for same supply or
profile hugely varies by study due to
– Genetic potential
– Management (2X vs. 3X milking, Health
condition)
– Environment (Season, Climate)
– Trial design
Modelization needed to reflect response as
%change of initial production
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Predicted Milk Yield % Change
Red plots are for studies with RP-LM (both Lys and Met) AND Blue plots are for studies with RP-L only (no Met supplementation)
Milk volume change depending MP-Lys grams/d Milk volume change depending Lys %MP
+0.5 point in Lys %MP
could result in+ 3.5 % in Milk Volume
+ 4 % in Milk Protein Yield
+ 2 % in Milk Fat Yield
+10 g MP-Lysine
could result in+ 3.5 % in Milk Volume
+ 4 % in Milk Protein Yield
+ 2 % in Milk Fat Yield
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Role of Methionine : look at trial with/without
0
0,01
0,02
0,03
0,04
0,05
0,06
Lys & Met Lys alone
+ Points
from Control
Milk protein % response
Milk protein content is primarily driven by Met
Lysine drives milk volume with little impact on %protein
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Maximal milk yield response to lysine in early lactation
24
Herd structure
• Robot milking system, 70 cows, average Days In Milk = 150
• Average production = 40,3 kg/cow, 3,25 % protein, 4,08 % fat
Test overview
• On Top 50g RP-Lysine (=12.5 g MP-Lysine)
• OFF/ON/OFF
• Analysis focusing on cows in early lactation.
No
supplementation
RP-Lysine
supplementationDifference
Early lactation
Cows 0-80 DIM43,5 45,7 +2,2 kg
Mid-End lactation
Average 200 DIM38,7 39,7 +1,0 kg
➔Response doubled for early lactation cows compared vs
others cows
Peak +4kg
+ 2,2 kg/cow during first 80 days
➔ Cows achieved higher peak during RP-Lysine
supplementation period
RP-Lysine
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Maximal milk yield response to lysine in early lactation
25
Spain, Robot milking 60 cows, Production = 35 kg/cow
Energy more limiting than Protein
+ lysine deficiency
OFF OFFONON OFF
Period All cowsCows
< 100 Days In Milk
OFF1 34.45 kg 38.6 kg
ON1 34.63 kg 40.7 kg
OFF2 33.40 kg 36.9 kg
ON2 34.50 KG 37.3 kg
OFF2 32.83 KG 36.7 kg
RP-Lysine effect +0.95 kg + 1.9 kg
No changes for %protein during ON periods vs OFF
New ration Crude Protein 16.05 %
+650g Corn grain
+33g RP-Lysine (=8.3 g MP-Lysine)
Switch Protein/Starch + RP-Lysine
-0.5kg Feed Robot (soybean)
-150g Rapeseed meal
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Models can still improve lysine & methionine requirements evaluation
Maximum benefits when both Lysine & Methionine are balanced (grams)
Some benefits to independant RP-Lysine or RP-Methio supplementation
In early lactation, extra lysine will support dry matter intake, milk volume and body
weight while extra methionine support liver function, energy status and
reproduction
RPAA Supplementation, independent and
additive in cattle
AA Concentrations, independent and additive in
mammary cell cultures
Mammary level modelling shows independent
and additive responses to energy and AA
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tureFormulating on Energy & AA for higher efficiency
Factors to improve nitrogen efficiency in lactating cattle
TotalDairy UK Seminar 2017
Optimize DMI & Maximize digestible forage intake
Energy supply
will determine
milk potential
of the ration
(ME allowable
milk)
Meet but not overly exceed the N requirements of rumen.
Keep N rumen supply
~120% requirement to
ensure >50% MP
supply by the rumen
Once you have ME and MP in balance and are happy with
rumen N balance, focus on
AA to meet requirements for ME allowable milk
Should formulate on Energy Corrected Milk
Maximize energy & rumen contribution then supply lysine & methionine to meet requirements
(grams) based on Energy allowable milk
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Recommendation for AA balancing during lactation
• Met 2.3-2.6 %MP
• Lys 6.8-7.2 %MP
• Lys/Met ratio 2.6 to 3
Optimum for AA profile
Calculating AA requirements based on energy supply
If ration supply = 55 Mcals ME
• Methionine req. = 55 mcal*1.10 g/Mcal = 60 g Met
• Lysine req. = 55 mcal*3,05 g/Mcal = 168 g Lys
→ Lys/Met = 2,8
• Don’t look at CP… focus on MP, RDP & AA
• Rumen Degradable Protein 10% DM
• Rumen N >120% requirements
• MP at 98-100% of requirements
Optimize protein supply
→ Mpreq [ 2 307 g – 2 609 g]
→ Mpreq [ 2 333 g – 2 470 g]
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For high producing cows, balance AA and Fatty Acids supply
31
Table developped for application in field by F.Gaudin – Big Gain, Mn
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Amino Acids have functions directly influencing cow milk potential
Nan, X., et al. (2014)
"Ratio of lysine to methionine
alters expression of genes
involved in milk protein
transcription and translation
and mTOR phosphorylation in
bovine mammary cells."
Physiological Genomics 46(7):
268-275.
Proliferation rate of Mammary Epithelial Cells is
increased by the supply of lysine & methionine.Combination of lysine and methionine as a
synergistic effect on proliferation rate.
• Lysine & methionine nutrition allow better expression of cow genetic potential by
maximizing udder capacity to synthetize milk protein
Milk yield at peak is determined by the number of Mammary Epithelial Cells in the udder
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Mobilization of AA from body protein during Transition Period
35
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Negative Protein Balance starts pre-calving
Robinson.P (2011). Productive responses of lactating dairy cattle to supplementing high levels
of ruminally protected lysine using a rumen protection technology. Animal Feed Science and
Technology 168 30– 41
“3-MH concentrations in plasma can be used to assess body protein synthesis and degradation (Blum et al., 1985),…
Starts earlier vs Negative Energy Balance
Is shorter vs NEB
Lysine is a major AA in body protein help udder preparation
support energy status
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Impact of Lysine on Intake & Body weight in early lactation
37
Schothorst Feed Research 2017, Transition Period trial
60 cows supplemented with RP-Lysine from 4 weeks pre-calving to12 weeks post-calving + 60 cows in Control group notsupplemented
DMI tend to increase faster after calving in the group supplemented with
RP-Lysine
After 3 weeks without supplementation, DMI came back to same level as
Control group
Supplementation RP-Lysine group
Stop
supplementation
Intake basal diet (PMR in Calent gate)
RP-Lysine group starts to regain weight 4 weeks after calving vs 8 weeks for
Control group
RP-Lysine group is back to calving weight at day 100 vs Control not yet back at
day 120Body weight monitoring
Impact of lysine on Body Condition Score
Robinson, P. H., et al. (2010). Productivity of lactating dairy cows as impacted by
feeding lysine in a ruminaly protected form. J. Dairy Sci. 93: 842.
“Output of milk (48.0 vs . 50.0 kg/cow/d), as wel l as milk fat , true
prote in and lactose, and energy, were higher (P < 0.05) in lysine
supplemented cows. In addit ion, the extent of body condit ion
score (BCS) loss was lower (P < 0.05) with Lys supplementat ion (−0.069
vs. −0.035 units/28 d)”
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Latest Ajinomoto research program about Lysine in Transition Period
38
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39
Exp.1 Effect of Lysine supplementation during prepartum
period on maternal DMI and milk production
Sugino et al., JDS. Vol. 101, Suppl. 2
Treatment MP, g/d mLys, g/d mMet, g/d
Control 1,143 80.5 26.0
Lysine 1,155 97.1 26.0
Animals
➢ 8 primiparous &
12 multiparous
Holstein cows
Con Control Control
Lys RP-Lysine
supplementation
Control
Measurements
➢ Daily DMI (Pre & Postpartum)
➢ Daily Milk Yield
➢ Weekly Milk Components
➢ Blood biomarkers
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Exp1. Results : DMI and Milk Production
✓ DMI increased pre-calving in Lysine supplemented group.
✓ DMI remained higher post-calving withoutsupplementation
40
Day
✓ Milk production increased in Lysine group.
Sugino et al., JDS. Vol. 101, Suppl. 2
Same diet for all cowsSame diet for all cows
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Exp.1 Results : Liver Functionality Index
41
0
20
40
60
80
100
120
140
160
-21 -14 0 7 21 35
T-CHO (mg/dL)
Control Lys
treatment: p=0.822
day: p<0.001treatment*day: p=0.546
Liver Functionality Index (LFI) Bertoni et al., 2013.Step 1 Albumin (ALB-I) subindex = 50 % 分娩後7日濃度 + 50 %(分娩後35日ー7日濃度)
Cholesterol (CHO-I) subindex = 50 % 分娩後7日濃度 + 50 %(分娩後35日ー7日濃度)Bilirubin (BIL-I) subindex = 67 % 分娩後7日濃度 + 333 %(分娩後7日ー35日濃度)
Step 2 LFI = (ALB-I ー 17.71)/1.08 + (CHO-I ー 2.57)/0.43 ー (BIL-I ー 6.08)/2.17
-3
-2
-1
0
1
2
3
Control Lys
LFI
treatment: p=0.10
LFI tended to be higher for Lysine group than
Control, indicating better liver functions in
spite of increased milk yield.
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Exp.2: Effect of RP-Lysine supplementation during prepartum and/or
postpartum period on maternal DMI and milk production
42
Test farm
Spruce Haven Research Center, NY
Animals
Cow N=109
Measurement
DMI, Milk production
Blood biomarkes
Con-Con Control Control
Lys-Con RP-Lysine Control
Con-Lys Control RP-Lysine
Lys-Lys RP-Lysine RP-Lysine
Pre-partum
Treatment MP, g/d Lys, g/d Met, g/d
RP-Lysine group 1238 99 26.2
Control group 1221 83 26.2
Recommendation 1400 97 32
Journal of Dairy Science vol.97, E-Suppl. 1: 328, 2014
Post-partum
• Keep supplementation during 3 first weeks.
• Monitor performances during 9 weeks
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Exp.2 Results
43
Milk production during 9 weeks
Test diets Control diet for all
DMI
Days
Impact of Lysine on Dry Matter Intake stops 3-4
weeks after end of supplementation
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Exp.2 Results : Blood BHBA Concentration
Con-Con
Con-Lys
High keton bodies concentration.
✓ BHBA concentration was suppressed in Lys-Con group indicating a better energy status
1200μmol/dL
1200μmol/dL1200μmol/dL
1200μmol/dL
Lys-Con
Lys-Lys
44
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45
C Control Control
L RP-Lysine supplementation
Control
Measurements
➢ Daily DMI (Pre & Postpartum)
➢ Daily Milk Yield
➢ Weekly Milk Components
➢ Insulin, GH, IGF-1
➢ Postpartum uterine health
Treatment MP, g/d mLys, g/d mMet, g/d
Control 1,037 69 29
RP-Lysine 1,058 89 29
Exp. 3 Kawashima et al. (to be published in 2020)
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46
Milk Production by Week
L (n=6)
C (n=6)Mean
Milk Y
ield
(kg
/d)
Week of lactation
Insulin (ng/ml)
L (n=6)
C (n=6)
L (n=6)
C (n=6)
C L
Milk, kg/d 42.8 46.5
• No DMI data is available yet.
• Cows fed Lys during pre-fresh showed
smoother take-off of milk production.
Blood hormones
• Pre-fresh Lys suppl. decreased
blood insulin, probably due to
improved insulin sensitivity?
• increased IGF-1 both pre- &
post-fresh, suggesting better
nutritional status & enhanced
GH-IGF axis
Exp. 3 Kawashima et al. (to be published in 2020)
L (n=6)
C (n=6)
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Exp. 3 Kawashima et al. (to be published in 2020)
47
Cytobrush
Pre-fresh Lys suppl.
• increased IgG concentration in the colostrum.
• indicating superior health & immune status of calves
from dams fed Lys during Pre-fresh.
Pre-fresh Lys suppl.
• tended to decrease the population of PMN
(polymorphonuclear) cells at 3 wks Post-fresh.
• indicating less inflammatory damages in the uterus
IgG conc. in the colostrum
103,2
n=21
121,3
n=22
0
20
40
60
80
100
120
140
C L
mg/ml
p=0.06
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Exp. 4 Cardoso et al., (2019, Unpublished)
C-C (n=18) Control Control
L-C (n=20) RP-Lysine Control
C-L (n=19) Control RP-Lysine
L-L (n=18) RP-Lysine RP-Lysine
Prepartum Postpartum
Treatment C L C L
MP, g/d 1,170 1,190 2,280 2,280
mLys, %MP 6.86 8.24 6.27 7.15
mLys, g/d 80.3 98.1 143 160
mMet, %MP 2.98 2.94 2.54 2.55
mMet, g/d 34.9 35.0 58.0 58.1
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Exp. 4 Cardoso et al., (2019, Unpublished)
0,0
0,5
1,0
1,5
2,0
11,0
11,5
12,0
12,5
13,0
C L
DMI Prepartum
DMI, kg DMI, %BW
Numerically higher for L, but
no significant difference
Prepartum L effect p = 0.08
Postpartum L effect p = 0.22
Body tissue mobilization right before calving
was suppressed by Lys supplementation.
Weekly change of pre-fresh BW
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Exp. 4 Cardoso et al., (2019, Unpublished)
50
Pre-fresh Lys supplementation;
• did not significantly affect colostrum quality
• resulted in insignificantly but numerically higher IgG Yield
• 20% more IgG in colostrum from cows fed Lys during pre -fresh
• Partially reproduced the result in Exp. 3 (Kawashima et al.)
Colostrum Quality
Pre-fresh Treatment P-value
Variable L C SEM PRE
Colostrum weight, kg 4.74 4.32 0.41 0.47
Colostrum IgG yield, g 434 367 36 0.20
Colostrum, % Brix 25.77 26.65 0.78 0.44
Colostrum fat, % 4.15 4.28 0.63 0.96
Colostrum true protein, % 16.87 17.65 0.57 0.42
Colostrum total solids 24.33 25.68 0.69 0.18
Data were collected at the first milking after calving (3.83 ± 3.63 h).
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Recommendation for lysine during close-up
Research studies tend to confirm that optimizing Lysine & Methionine in Close Up Dry diets
help
Increase DMI and milk production of
mother cows. Production response diminish if
post-partum diet not well balanced
Improve health & nutrition
status of mother cows.
Improve health & nutrition
status of newborn calves.
Example; if your close-up cows are consuming
13 kg/d DMI...
MP, g/d mLys, g/d
Formulation
Target1,300 > 90
• Metabolizable Proteine: 100 g MP / kg DMI
• mLysine: > 90 g/d
Practical targets in the close-up
cow ration
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Conclusion, key messages
52
Lysine is an essential amino acid for
dairy cows. MP-Lysine is good
predictor of milk protein yield
Grams of MP-Lysine will determine
production potential while Lysine
%MP will drive conversion
efficiency
Microbial Protein is the best source
of MP-Lysine. Maximise RD-CHO
for best Microbial MP-Lysine
production
Balance lysine with methionine and
energy for optimal efficiency
Lysine in early lactation drives milk
volume with little effect on
%protein.
Cow response to lysine is maximum
in early lactation (x2 compared to
« All herd »)
For optimal response to lysine
supplementation target herds <150
Days In Milk
Lysine nutrition in close-up impact
dry matter intake post-calving, and
milk production
Lysine nutrition during close-up
impact cow’s health status and also
calves (colostrum IgG, colostrum
absorption capacity)