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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 True protein Saliva NPN Urea Ammonia Liver Peptides RUMEN Amino acids RUP Amino acids Microbial protein SMALL INT RUP Microbial protein Endogenous protein Metabolizable protein (absorbed AA) Mammary gland MILK Amino acid supply and use in dairy cows Amino Acids: The Required Nutrients FOR THE COW Essential 1. 2. 3. 4. 5. 6. 7. 8. 9. 10. Arginine Histidine Isoleucine Leucine Lysine Methionine Phenylalanine Threonine Tryptophan Valine Non-essential 1. 2. 3. 4. 5. 6. 7. 8. 9. 10. Alanine Aspartic acid Asparagine Cysteine Glutamic acid Glutamine Glycine Proline Serine 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? Amino acid Rulquin (2001) Doepel et al. (2004) Segmented Logistic 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 Corn gluten meal 1.7 2.4 2.1 Cotton seed 4.3 1.7 2.8 Ideal 7.2 2.5 2.5 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 Optimum content of Lys in MP 0.15 7.2 Milk protein content responses, g/100 g 0.10 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 Percent Lys in MP (Met > 1.95 of MP) 8.4 8.8 9.2 9.6 10.0 Optimum content of Met in MP 0.20 Milk protein content responses, (g/100 g) 2.4 0.15 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 Percent Met in MP (Lys > 6.50 of MP) 3.20 3.40 Current knowledge regarding optimum AA concentrations in MP 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 Model 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 Organic acids Sugars Starches Cell walls Fructans (glucose, fructose, sucrose, lactose) Pectic substances and B-glucan Hemicellulose Cellulose NDSF NDSC (NFC) ADF NDF Hall, 1999 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 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) 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 Current knowledge regarding optimum AA concentrations in MP 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 Model Whitehouse et al. (2013) 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 Brewer’s grains 4.1 Milk 7.7 Canola meal 5.6 Bacteria 7.9 Corn DDGS 2.2 Corn gluten feed 2.7 Corn gluten meal 1.7 Cotton seed 4.3 Ideal 7.2 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 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 Current knowledge regarding optimum AA concentrations in MP 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 Model Whitehouse et al. (2013) 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 Corn gluten meal 1.7 2.4 Cotton seed 4.3 1.7 Ideal 7.2 2.5 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 Met supplements AjiPro-L Smartamine M® LysiPEARL Mepron® M85 Lysine 35 AminoShure-M Megamine-L MetiPEARL MetaboLys USA Lysine MetaSmart® (HMBi) Alimet® Rhodimet AT 88® MFP™ (CaMHA) 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 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 MP1 (%) MP Microbial required1 MP1 (g/d) (g/d) Endog. MP1 (g/d) Required MP from Required Required RUP RUP2 RUP3 (g/d) (g/d) (% 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 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 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 Corn gluten meal 1.7 2.4 2.1 Cotton seed 4.3 1.7 2.8 Ideal 7.2 2.5 2.4 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 RDP 9.8 9.1 Roasted whole soybeans 5.5 6.6 RUP 5.9 4.5 Canola meal, mechanically extracted 5.0 3.0 CP 15.7 13.6 Soy Plus 5.5 0.5 Molasses 4.2 4.2 AMP DMP % of DM 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 38.8a 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 <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 13.0a 10.3bc 10.1c 11.1b <0.01 BUN, mg/dl 11.5a 6.8b 7.6b 8.0b <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 2.50 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 4.10 3.90 3.70 3.50 3.30 3.10 2.90 2.70 2007 2008 2009 Year - Month 2010 2011 Fat Protein Benefits of increasing Lys and Met in MP to more adequate levels 1) 2) 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 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) 2) 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! Benefits of increasing Lys and Met in MP to more adequate levels 1) 2) 3) 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 Benefits of increasing Lys and Met in MP to more adequate levels 1) 2) 3) 4) 5) 6) 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 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. 2. 3. 4. 5. 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