1. Advances in Protein and Amino Acid Nutrition: Implications on Transition Cow Performance
Chuck Schwab
Schwab Consulting, LLC, Boscobel, WI
Professor Emeritus, Animal Sciences
University of New Hampshire

2. Protein Metabolism in Ruminants
Crude protein
Saliva
True protein
Peptides
Amino
acids
Ammonia
NPN
Microbial protein
Urea
Liver
Microbial
protein
RUMEN
SMALL
INT
RUP
Endogenous
Metabolizable protein (absorbed AA)
Mammary
gland
MILK

3. Amino acid supply and use in dairy cows

4. Amino Acids: The Required Nutrients FOR THE COW
Essential
Arginine
Histidine
Isoleucine
Leucine
Lysine
Methionine
Phenylalanine
Threonine
Tryptophan
Valine
Non-essential
Alanine
Aspartic acid
Asparagine
Cysteine
Glutamic acid
Glutamine
Glycine
Proline
Serine
Tyrosine

5. Functions of Amino Acids
Required building blocks for the synthesis of tissue, regulatory, protective and secretory proteins…100’s are synthesized every day
The AA composition of each protein is different
Protein synthesis is a genetically determined event; i.e., AA composition of a protein is the same every time it is synthesized
AA are key regulators of various pathological and physiological processes, including immune responses
AA are also used to synthesize all of the other N-containing compounds in the body [e.g., dozens of compounds such as hormones, neurotransmitters, nucleotides (RNA and DNA), histamine, polyamines (e.g., spermine and spermidine), etc.]

6. What is the Ideal Balance of Absorbed Amino Acids?
Rulquin (2001)
Doepel et al. (2004)
Segmented Logistic
linear model model
Arginine
3.1
4.8
4.6
Histidine*
3.0
2.4
Isoleucine
4.5
5.3
Leucine
8.9
9.4
Lysine*
7.3
7.2
Methionine*
2.5
Phenylalanine
5.2
5.5
Threonine
4.0
5.1
5.0
Tryptophan
1.7
Valine
6.1
6.5

7. Lys and Met in lean tissue, milk, rumen bacteria and feedstuffs (% of protein), relative to predicted optimum concentrations in MP
Lys
Met
His
Tissue
6.3
1.8
2.4
Brewer’s grains
4.1
1.7
2.0
Milk
7.7
2.7
Canola meal
5.6
1.9
2.8
Bacteria
7.9
2.6
Corn DDGS
2.2
2.5
Corn gluten feed
1.6
2.9
Ideal
7.2
Corn gluten meal
2.1
Cotton seed
4.3
Alfalfa silage
4.4
1.4
Linseed meal
3.7
Corn silage
1.5
Soybean meal
Grass silage
3.3
1.2
Blood meal
9.0
6.4
Barley
3.6
2.3
Feather meal
0.8
Corn
3.1
Fish meal
Wheat
Meat meal
5.4

8. Limiting AA Theory
First limiting AA = the essential AA supplied in the smallest amount relative to requirements
Second limiting AA = the essential AA supplied in the second smallest amount relative to requirements

9. Optimum content of Lys in MP

10. Optimum content of Met in MP

11. Current knowledge regarding optimum AA concentrations in MP
Model
Lysine
Methionine
Optimal Lys/Met ratio
NRC (2001), revised
6.83
2.28
3.00
CPM-Dairy
7.46
2.57
2.90
AMTS v.3.3.4
6.97
2.53
2.75
Whitehouse et al. (2013)

12. Goal : To meet RDP and RUP requirements for optimum performance with minimum amounts of each
1) RDP – purpose is to meet the ammonia and AA requirements of rumen microbes for maximum carbohydrate digestion and synthesis of microbial protein
2) RUP – purpose is to provide the additional AA, in the correct balance, that the cow requires that are not provided by microbial protein

13. Practical Protein and Amino Acid Balancing Guidelines
6 steps
1. Feed a blend of high quality fermentable feeds and physically effective fiber to maximize synthesis of VFA and microbial protein

14. Right blend of carbohydrates?
PLANT CARBOHYDRATES
Cell contents
Cell
walls
Organic
acids
Sugars
(glucose,
fructose,
sucrose,
lactose)
Starches
Fructans
Pectic substances and
B-glucan
Hemicellulose
Cellulose
NDSF
ADF
NDSC (NFC)
NDF
Hall, 1999

15. Practical Amino Acid balancing Guidelines
6 steps
1. Feed a blend of high quality fermentable feeds and physically effective fiber to maximize synthesis of VFA and microbial protein
2. Feed adequate but not excessive levels of RDP to meet rumen bacterial requirements for AA and ammonia to allow for maximum CHO digestion and synthesis of microbial protein

16. Factors affecting RDP requirements
1) Intake and mixture of fermentable carbohydrates
2) Quality of RDP (relative supplies of protein, free AA and ammonia and rate of degradation)
Fermentable carbohydrates
RDP
Microbial protein
VFA’s

17. Practical Amino Acid balancing Guidelines
6 steps
1. Feed a blend of high quality fermentable feeds and physically effective fiber to maximize synthesis of VFA and microbial protein
2. Feed adequate but not excessive levels of RDP to meet rumen bacterial requirements for AA and ammonia to allow for maximum CHO digestion and synthesis of microbial protein
3. Feed high-Lys protein supplements or a combination of high-Lys protein supplements and a RPLYS supplement to achieve a level of Lys in MP that comes close to meeting the optimal concentration

18. Current knowledge regarding optimum AA concentrations in MP
Model
Lysine
Methionine
Optimal Lys/Met ratio
NRC (2001), revised
6.83
2.28
3.00
CPM-Dairy
7.46
2.57
2.90
AMTS v.3.3.4
6.97
2.53
2.75
Whitehouse et al. (2013)

19. Lys and Met in lean tissue, milk, rumen bacteria and feedstuffs (% of protein), relative to predicted optimum concentrations in MP
Lys
Met
His
Tissue
6.3
Brewer’s grains
4.1
Milk
7.7
Canola meal
5.6
Bacteria
7.9
Corn DDGS
2.2
Corn gluten feed
2.7
Ideal
7.2
Corn gluten meal
1.7
Cotton seed
4.3
Alfalfa silage
4.4
Linseed meal
3.7
Corn silage
2.5
Soybean meal
Grass silage
3.3
Blood meal
9.0
Barley
3.6
Feather meal
2.6
Corn
2.8
Fish meal
Wheat
Meat meal
5.4

20. Commercially available RP-Lys supplements in the United States
AjiPro-L
AminoShure-L
LysiPEARL
Lysine 35
Megamine-L
MetaboLys
USA Lysine

21. Practical Amino Acid balancing Guidelines
6 steps
1. Feed a blend of high quality fermentable feeds and physically effective fiber to maximize synthesis of VFA and microbial protein
2. Feed adequate but not excessive levels of RDP to meet rumen bacterial requirements for AA and ammonia to allow for maximum CHO digestion and synthesis of microbial protein
3. Feed high-Lys protein supplements or a combination of high-Lys protein supplements and a RPLYS supplement to achieve a level of Lys in MP that comes close to meeting the optimal concentration
4. Feed a “rumen-protected” Met supplement in amounts “needed” to achieve optimum Lys/Met ratio in MP…then fine tune for maximal milk protein concentrations

22. Current knowledge regarding optimum AA concentrations in MP
Model
Lysine
Methionine
Optimal Lys/Met ratio
NRC (2001), revised
6.83
2.28
3.00
CPM-Dairy
7.46
2.57
2.90
AMTS v.3.3.4
6.97
2.53
2.75
Whitehouse et al. (2013)

23. Lys and Met in lean tissue, milk, rumen bacteria and feedstuffs (% of protein), relative to predicted optimum concentrations in MP
Lys
Met
His
Tissue
6.3
1.8
Brewer’s grains
4.1
1.7
Milk
7.7
2.7
Canola meal
5.6
1.9
Bacteria
7.9
2.6
Corn DDGS
2.2
Corn gluten feed
1.6
Ideal
7.2
2.5
Corn gluten meal
2.4
Cotton seed
4.3
Alfalfa silage
4.4
1.4
Linseed meal
3.7
Corn silage
1.5
Soybean meal
Grass silage
3.3
1.2
Blood meal
9.0
Barley
3.6
Feather meal
0.8
Corn
2.8
2.1
Fish meal
Wheat
Meat meal
5.4

24. Commercially available ruminant AA supplements in the United States
Lys supplements
Met supplements
AjiPro-L
LysiPEARL
Lysine 35
Megamine-L
MetaboLys
USA Lysine
Smartamine M®
Mepron® M85
AminoShure-M
MetiPEARL
MetaSmart® (HMBi)
Alimet®
Rhodimet AT 88®
MFP™ (CaMHA)

25. Practical Amino Acid balancing Guidelines
6 steps
1. Feed a blend of high quality fermentable feeds and physically effective fiber to maximize synthesis of VFA and microbial protein
2. Feed adequate but not excessive levels of RDP to meet rumen bacterial requirements for AA and ammonia to allow for maximum CHO digestion and synthesis of microbial protein
3. Feed high-Lys protein supplements or a combination of high-Lys protein supplements and a RPLYS supplement to achieve a level of Lys in MP that comes close to meeting the optimal concentration
4. Feed a “rumen-protected” Met supplement in amounts “needed” to achieve optimum Lys/Met ratio in MP…then fine tune for maximal milk protein concentrations
5. Limit RUP supplementation to what the “cows say” is needed…reductions of 1 to 2 percentage units of DM are common

26. How much supplemental RUP do you feed?
Factors affecting RUP requirements (all of which we never quite know):
Intestinal supply of microbial protein
RUP digestibility
RUP-Lys digestibility*
Concentrations of Lys and Met in MP*

27. Effect of Lys and Met in MP on amounts of MP and RUP required to provide 180 g MP-Lys and 60 g MP-Met
Lys in MP1
(%) MP
required1
(g/d)
Microbial MP1
Endog.
MP1
Required MP from RUP
Required
RUP2
Required RUP3
(% DM)
5.7/1.9
3157
1390
121
1646
2058
8.1
6.0/3.0
3000
1489
1861
7.3
6.3/2.1
2857
1346
1683
6.6
6.6/2.2
2727
1216
1520
6.0
6.9/2.3
2609
1098
1372
5.4
1 NRC (2001) was used as model of choice. Ration was balanced for 40.9 kg of 3.2% protein milk. MP required = 2857 g
2 Assumed an average RUP digestibility of 80%
3 DM intake assumed to be 25.5 kg

28. Practical Amino Acid balancing Guidelines
6 steps
1. Feed a blend of high quality fermentable feeds and physically effective fiber to maximize synthesis of VFA and microbial protein
2. Feed adequate but not excessive levels of RDP to meet rumen bacterial requirements for AA and ammonia to allow for maximum CHO digestion and synthesis of microbial protein
3. Feed high-Lys protein supplements or a combination of high-Lys protein supplements and a RPLYS supplement to achieve a level of Lys in MP that comes close to meeting the optimal concentration
4. Feed a “rumen-protected” Met supplement in amounts “needed” to achieve optimum Lys/Met ratio in MP…then fine tune for maximal milk protein concentrations
5. Limit RUP supplementation to what the “cows” say is needed…reductions of 1 to 2 percentage units of DM are common
6. Monitor His levels in MP closely

29. Lys and Met in lean tissue, milk, rumen bacteria and feedstuffs (% of protein), relative to predicted optimum concentrations in MP
Lys
Met
His
Tissue
6.3
1.8
2.4
Brewer’s grains
4.1
1.7
2.0
Milk
7.7
2.7
Canola meal
5.6
1.9
2.8
Bacteria
7.9
2.6
Corn DDGS
2.2
2.5
Corn gluten feed
1.6
2.9
Ideal
7.2
Corn gluten meal
2.1
Cotton seed
4.3
Alfalfa silage
4.4
1.4
Linseed meal
3.7
Corn silage
1.5
Soybean meal
Grass silage
3.3
1.2
Blood meal
9.0
6.4
Barley
3.6
2.3
Feather meal
0.8
Corn
3.1
Fish meal
Wheat
Meat meal
5.4

30. Is His limiting after Lys and Met?
Ingredients
Adequate MP
Deficient MP
Corn silage
40.2
Alfalfa haylage
16.7
16.6
Grass hay
5.8
Cottonseed hulls
1.1
Corn grain, ground
5.7
11.7
Bakery by-product meal
7.4
Roasted whole soybeans
5.5
6.6
Canola meal, mechanically extracted
5.0
3.0
Soy Plus
0.5
Molasses
4.2
AMP
DMP
% of DM
RDP
9.8
9.1
RUP
5.9
4.5 CP
15.7
13.6
Lee et al. (2012)

31. Is His limiting after Lys and Met?
AMP
DMP
DMPLM
DMPLMH
P value
DM intake, kg/d
24.5
23.0
23.7
24.3
0.06
Milk, kg/d
38.8a
35.2b
36.9ab
38.5a
<0.01
Milk protein, %
2.98
2.94
2.99
3.03
0.23
Milk protein, g/d
1130a
1010b
1100a
1140a
Milk fat, %
3.50
3.51
3.32
3.30
0.44
Milk fat, g/d
1340
1200
1210
1230
0.10
MUN, mg/dl
13.0a
10.3bc
10.1c
11.1b
BUN, mg/dl
11.5a
6.8b
7.6b
8.0b
Lee et al. (2012)

32. Benefits of increasing Lys and Met in MP to more adequate levels
1) Increased milk component concentrations
No longer uncommon to hear reports of increases in milk protein concentrations of 0.20 to 0.25 percentage units and increases in milk fat concentrations of 0.10 to 0.15 percentage units…often on less dietary RUP.
Increases in milk protein percentages are the most visible of the responses to better AA nutrition…”the tip of the iceberg”!
You don’t have to accept low components because of high production

34. Benefits of increasing Lys and Met in MP to more adequate levels
Increased milk component concentrations
Increased milk yield…particularly in early lactation cows
Early studies indicated 2 to 5 lb more milk in early lactation…more recent studies have shown 5 to 10 lb more milk

35. Milk yield responses to feeding RP-Lys or RP-Lys + Met to early lactation cows in 15 experiments

36. Benefits of increasing Lys and Met in MP to more adequate levels
Increased milk component concentrations
Increased milk yield…particularly in early lactation cows
Early studies indicated 2 to 5 lb more milk in early lactation…more recent studies have shown 5 to 10 lb more milk
Increased milk yields in early lactation may or may not be accompanied by increases in milk protein percentages if levels of Lys and Met in MP are not pushed high enough
If you see an increase in milk protein percentage, assume at least some increase in milk yield!

37. Benefits of increasing Lys and Met in MP to more adequate levels
Increased milk component concentrations
Increased milk yield…particularly in early lactation cows
Reduced requirement for RUP for similar or higher milk milk component levels and milk yields

38. Benefits of increasing Lys and Met in MP to more adequate levels
Increased milk component concentrations
Increased milk yield…particularly in early lactation cows
Reduced requirement for RUP for similar or higher milk milk component levels and milk yields
More predictable changes in milk production to changes in RUP supply
Less metabolic disorders
INCREASED HERD PROFITABILITY

39. Transition Cows 1) Fat and protein mobilization (and other nutrients)
Feed intake does not keep pace with nutritional needs
Some important metabolic changes:
1) Fat and protein mobilization (and other nutrients)
2) BW loss and usually dramatic increases in plasma NEFA
3) Increased uptake of FA by the liver (often in amounts that exceed capacity for oxidation)
4) Increased ketone production (ketosis)
5) Increased storage of TG in the liver (fatty liver)
6) Reduced liver function (e.g., depressed glucose production)
7) Increased inflammation [characterized by an increase in production of posAPP (e.g., haptoglobin and serum amyloid A) and a decrease in the production of negAPP (e.g., albumin)]. The trigger for these responses are the pro-inflammatory cytokines (e.g., IL-6, IL-1 and TNF-α)
8) Increased oxidative stress…the result of an imbalance between production of ROM and the neutralizing capacity of antioxidant mechanisms

40. Negative impacts of ketosis and fatty liver
Increased risk for herd removal (3x) (McArt et al., 2012)
Increased risk for displaced abomasum (2.6x to 19.3x in 3 studies) (early detection and treatment of subclinical ketosis with oral propylene glycol reduces risk)
Increased risk for metritris (2.3x to 3.4x in 2 studies)
Impaired fertility – results are inconsistent (early detection and treatment of subclinical ketosis with oral propylene glycol increases first service conception)
Obvious economic impact on herd profitability!
Oetzel, 2012

41. Fatty Liver
50-60% of cows experience moderate to severe fatty liver…peak fat content is about 10 days after calving
“Fatty liver is a classic symptom for choline deficiency, therefore it is reasonable to question if transition cows are typically deficient in choline” (Grummer, 2012)
Question: Might it also be reasonable to question if a Met deficiency, or AA deficiencies in general, contribute to fatty livers since protein synthesis is a fundamental initial step to virtually every metabolic reaction?

42. Findings regarding protein metabolism of transition cows
The RUP requirements of post-fresh transition cows are higher than at any other time within their lactation
An evaluation of dry cow diets with NRC (2001) and research experiments both indicate most post-fresh transition cows within a herd will experience deficiencies of MP (100 to 600 g/d) (3 to 15 kg milk)
Shortages of MP are extremely variable among transition cows (contributing factors are on DIM, DMI and milk yield, health status, etc.)
Conclusion: First 2-3 wk of lactation clearly challenges the AA status of the cow. So…the question is “How important is it to balance diets of transition cows for AA?”

43. Benefits of higher levels of Lys and Met in MP for transition cows
Higher DM intake
More milk and higher protein milk
Energy balance and NEFA usually not affected
However, evidence exists the liver is healthier:
Fat doesn’t accumulate as fast (Osorio et al., 2013)
Evidence of greater VLDL synthesis (Bauchart et al., 1992; Osorio et al., 2013)
Pathways associated with carbohydrate metabolism are significantly impacted (e.g., gluconeogenesis) (Osorio et al., 2013)
Decreased plasma ceruloplasmin and serum amyloid A, and increased total antioxidants and glutathione (antioxidant) (Osorio et al., submitted)
Met supplementation, in the presence of high Lys, appears to reduce the inflammatory state after calving

44. Milk yield responses to feeding RP-Lys or RP-Lys + Met to early lactation cows in 15 experiments

45. Benefits of higher levels of Lys and Met in MP for transition cows
Higher DM intake
More milk and higher protein milk
Energy balance and NEFA usually not affected
New evidence indicates the liver and cow are healthier:
Fat doesn’t accumulate as fast (Osorio et al., 2013)
Greater VLDL synthesis (Bauchart et al., 1992; Osorio et al., 2013)
Pathways associated with carbohydrate metabolism are significantly impacted (e.g., gluconeogenesis) (Osorio et al., 2013)
Decreased plasma ceruloplasmin and serum amyloid A, and increased total antioxidants and glutathione (antioxidant) (Osorio et al., submitted)
These studies indicate that Met supplementation, in the presence of high Lys, enhanced the antioxidant capacity and reduced the inflammatory signaling in the liver

46. Advances in Protein and Amino Acid Nutrition
Diet evaluation and ration formulation models are getting better in predicting RDP, RUP and AA supplies to the cow
NRC (2001) and Formulate II
Spartan
AMTS.Cattle and NDS Professional (both use CNCPS v.6.1 biology)
Nittany Cow Ration Evaluator
Feed testing labs are providing more information
Estimates of digestibility (NDF, starch, RUP, etc.)
More chemical components (lactic acid, VFA, sugars, AA, etc.)
More commercial sources of RP-Met and Lys supplements
Only way to meet “really” meet Met requirements
Easier to meet Lys requirements
Gaining appreciation for differences in efficacy of products
More precise protein nutrition healthier cows + more milk + higher component milk + lower CP feeding

47. Summary and Conclusions
Protein nutrition has evolved from balancing rations for CP to balancing for RDP, RUP and AA in MP
Cows require AA for tissue and protein synthesis 
AA balancing is important for optimizing usage of “bypass protein” and maximizing milk and milk component synthesis, transition cow health, and dairy herd profitability
Choose protein and AA supplements carefully
As expected, health and production impacts are greatest in early lactation cows, whereas benefits on RUP sparing are greatest in cows after peak DM intake