Advances in Protein and Amino Acid Nutrition: Implications on Transition Cow Performance

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

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

Amino acid supply and use in dairy cows

Amino Acids: The Required Nutrients FOR THE COW Essential 1. Arginine 2. Histidine 3. Isoleucine 4. Leucine 5. Lysine 6. Methionine 7. Phenylalanine 8. Threonine 9. Tryptophan 10. Valine Non-essential 1. Alanine 2. Aspartic acid 3. Asparagine 4. Cysteine 5. Glutamic acid 6. Glutamine 7. Glycine 8. Proline 9. Serine 10. Tyrosine

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.]

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

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

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

Milk protein content responses, g/100 g Optimum content of Lys in MP 0.15 0.10 7.2 0.05 0.00-0.05-0.10-0.15-0.20-0.25 4.4 4.8 5.2 5.6 6.0 6.4 6.8 7.2 7.6 8.0 8.4 8.8 9.2 9.6 10.0 Percent Lys in MP (Met > 1.95 of MP)

Milk protein content responses, (g/100 g) Optimum content of Met in MP 0.20 0.15 2.4 0.10 0.05 0.00-0.05-0.10-0.15-0.20 1.60 1.80 2.00 2.20 2.40 2.60 2.80 3.00 3.20 3.40 Percent Met in MP (Lys > 6.50 of MP)

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)

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

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

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

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

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 6.3 Grass silage 3.3 Blood meal 9.0 Barley 3.6 Feather meal 2.6 Corn 2.8 Fish meal 7.7 Wheat 2.8 Meat meal 5.4 Lys Met His

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

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

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

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

How much supplemental RUP do you feed? Factors affecting RUP requirements (all of which we never quite know): 1) Intestinal supply of microbial protein 2) RUP digestibility 3) RUP-Lys digestibility* 4) Concentrations of Lys and Met in MP*

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 MP 1 (%) MP required 1 (g/d) Microbial MP 1 (g/d) Endog. MP 1 (g/d) Required MP from RUP (g/d) Required RUP 2 (g/d) Required RUP 3 (% DM) 5.7/1.9 3157 1390 121 1646 2058 8.1 6.0/3.0 3000 1390 121 1489 1861 7.3 6.3/2.1 2857 1390 121 1346 1683 6.6 6.6/2.2 2727 1390 121 1216 1520 6.0 6.9/2.3 2609 1390 121 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

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

Is His limiting after Lys and Met? Ingredients Adequate MP Deficient MP Corn silage 40.2 40.2 Alfalfa haylage 16.7 16.6 Grass hay 5.8 5.8 Cottonseed hulls 1.1 1.1 Corn grain, ground 5.7 11.7 Bakery by-product meal 7.4 7.4 Roasted whole soybeans 5.5 6.6 Canola meal, mechanically extracted 5.0 3.0 Soy Plus 5.5 0.5 Molasses 4.2 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)

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 38.8 a 35.2 b 36.9 ab 38.5 a <0.01 Milk protein, % 2.98 2.94 2.99 3.03 0.23 Milk protein, g/d 1130 a 1010 b 1100 a 1140 a <0.01 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 13.0 a 10.3 bc 10.1 c 11.1 b <0.01 BUN, mg/dl 11.5 a 6.8 b 7.6 b 8.0 b <0.01 Lee et al. (2012)

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

April May June July August Sept Oct Nov Dec Jan Feb Mar April May June July Aug Sept Oct Nov Dec Jan Feb Mar April May June July Aug Sept Oct Nov Dec Jan Feb Mar April May June July August Sept Oct Nov Dec Jan Feb Mar April May June July Aug Sept Percent Monthly Average Fat and Protein % - Wisconsin Dairy Fat 4.10 Protein 3.90 3.70 3.50 3.30 3.10 2.90 2.70 2.50 2007 2008 2009 2010 2011 Year - Month

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

Benefits of increasing Lys and Met in MP to more adequate levels 1) Increased milk component concentrations 2) 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!

Benefits of increasing Lys and Met in MP to more adequate levels 1) Increased milk component concentrations 2) Increased milk yield particularly in early lactation cows 3) Reduced requirement for RUP for similar or higher milk milk component levels and milk yields 4) More predictable changes in milk production to changes in RUP supply 5) Less metabolic disorders 6) INCREASED HERD PROFITABILITY

Transition Cows 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

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

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?

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?

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

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

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

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

Summary and Conclusions 1. Protein nutrition has evolved from balancing rations for CP to balancing for RDP, RUP and AA in MP 2. Cows require AA for tissue and protein synthesis 3. AA balancing is important for optimizing usage of bypass protein and maximizing milk and milk component synthesis, transition cow health, and dairy herd profitability 4. Choose protein and AA supplements carefully 5. 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