Lysine as an amino acid in ruminants: where are we ?Matthieu Rolland; Ajinomoto Animal Nutrition Europe

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Where do I come from ? Background in dairy

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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

FOUNDATIONS – WHY SHOULD WE CARE ABOUT LYSINE ?

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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.

SOURCES OF METABOLIZABLE LYSINE FOR DAIRY COWS

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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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Watch out digestibility of RUP sources

12

LYSINE REQUIREMENTS AND CONVERSION EFFICIENCY

(COMPONENTS)

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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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18

Lysine

DAIRY COW RESPONSE TO LYSINE

META ANALYSIS (VOLUME)

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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

GUIDELINES TO BALANCE

RATION FOR LYSINE, METHIONINE

AND ENERGY

26Sources :

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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

LYSINE FUNCTIONS AND

RECOMMENDATIONS DURING

TRANSITION PERIOD

32Sources :

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33

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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)