1. Protein Metabolism II ANS 520

3. Protein Pathways

4. Fate of Rumen Ammonia 1. Bacterial protein synthesis 2. Absorbed from reticulorumen and omasum NH 3 passes from rumen by diffusion into portal blood. (High concentration to low) Form of ammonia dependent on pH of rumen NH 3 + H + NH 4 + Less absorption at more acid pH 3. At pH of rumen, no NH 3 lost as gas

5. Fate of Absorbed Ammonia 1. Transported to liver by portal vein 2. Converted to urea via urea cycle in liver NH 3 Urea Urea cycle 3. Urea released into blood 4. If capacity of urea cycle in liver is exceeded Ammonia toxicity Over consumption of urea

6. Fate of Blood Urea 1. Excreted into urine 2. Recycled to digestive tract, g N/d Saliva – Related to concentration of urea in blood Sheep: 0.5 to 1.0 Cattle: 1.0 to 7.6 Diffusion into GIT Sheep: 2 to 5 Cattle: 25 to 40

7. Urea Diffusion into Rumen Rumen wall Blood urea Urea NH 3 Bacterial population 1.Total N transferred is greater when high N diets are fed. 2.Percentage of diet N transferred is greater when low N diet are fed

8. Urea Diffusion into Rumen Update Rumen wall Urea transporter Blood urea Urea High [NH 3 ] inhibits NH 3 Bacterial population

9. Adjustments to Low Protein Intake Kidney Blood urea Urea Urine urea Urea is predominant form of N in urine Reabsorption of urea by kidney increased when ruminants fed low N diets Conserves nitrogen in the body Greater portion recycled to digestive tract Sheep fed the same diet tend to reabsorb more urea than cattle

10. Nitrogen Recycling - Cattle Marini et al. JAS 2003

11. Sources of Nitrogen Recycled to GIT 1.Urea flowing back into digestive tract  Rumen Saliva Diffusion from blood  Lower digestive tract (large intestine, colon, cecum) Diffusion from blood Endogenous protein secretions into GIT  Mucins  Enzymes  Sloughing of tissue 2.Turnover of microbial cells in rumen & reticulum

12. Significance of Recycled Nitrogen Source of N for microbes when protein consumption is limited Wild species Protein intake during winter is very low Rumen deficient of nitrogen for microbial activity Slowly degraded feed proteins Recycling provides nitrogen for microbial growth Infrequent feeding of supplemental protein Programs to reduce supplemental nitrogen Difficult to make ruminants severely protein deficient

13. Urea Nitrogen - Cattle Marini et al. JAS 2003

14. Amino Acid Synthesis Ammonia Fixation 1. Glutamine synthetase/glutamate synthase Glutamine synthetase Glu + NH 3 + ATPGln Glutmate synthase  -ketoglutarate + glutamine + NADPH 2 2 Glu High affinity for NH 3 - Concentrates NH 3 in cells – Uses ATP Because of N recycling this reaction may not be that important

15. Amino Acid Synthesis Ammonia Fixation 2. Glutamic dehydrogenase  -ketoglutarate + NH 3 + NADHGlu Low affinity for NH 3 – High concentration of enzyme in rumen bacteria – Does not use ATP Probably predominant pathway 3. Other AA can be synthesized by transamination reactions with glutamic acid Estimates of NH 3 requirements range from 5 (culture) to 20 mg/100 ml (in situ digestion)

16. Amino Acid Composition % Crude Protein or G/100g CP TissueMilk ----------Bact ----------CornSoy Cell wallNon wallMean Methionine1.972.712.402.682.602.281.46 Lysine6.377.625.608.207.903.036.32 Histidine2.472.741.742.692.003.162.72 Phenylalanine3.534.754.205.165.105.325.65 Tryptophan0.491.51NA1.63-0.891.46 Threonine3.903.723.305.595.803.674.18 Leucine6.709.185.907.518.1012.667.95 Isoleucine2.845.794.005.885.703.675.44 Valine4.035.894.706.166.205.325.65 Arginine3.303.403.826.965.105.067.53

17. Amino Acids in Undegraded Feed Proteins HisIslLysMet Fish meal3.44.26.63.1 Fish meal residue2.94.96.02.9

18. Sources of Amino Acids for Host Animal 1. Microbial proteins Quantity determined by: a)Fermentability of the feed b)Quantity of feed consumed c) Nitrogen available to microorganisms 2. Undegraded feed proteins (UIP) Quantity will vary in relation to: a) Degradability of feed proteins b) Quantity of feed proteins consumed

19. Nutritional Value of Microbial Proteins 1996 NRC for Beef Microbial protein 80% digestible in the intestine UIP 80% digestible in the intestine 2001 NRC for Dairy and Level 1 CNCPS Microbial protein 80% digestible in the intestine Digestibility of RUP (UIP) is variable in Dairy NRC UIP 80% digestible in Level 1 CNCPS

20. History of Protein Systems for Ruminants ISU Metabolizable protein system Wisconsin system – When urea could be used Several European systems – Mostly MP systems 1985 NRC system – Summarized systems & Proposed a MP system Used in 1989 Dairy NRC Cornell CNCPS 1996 Beef NRC system – Mostly CNCPS system Used in ISU Brands system 2001 Dairy NRC system

21. NH 3 Blood urea Urine Amino acid pools Energy NH 3 Metabolizable Microbial protein protein Protein Protein from diet Rumen Intestine Feces A B C Metabolizable Protein Model Tissue proteins

22. Protein Metabolism of Ruminants Concept of Metabolizable Protein Metabolizable protein (MP) = Absorbed amino acids or = Digestible fraction of microbial proteins + digestible fraction of undegraded feed proteins Digestible protein (amino acids) available for metabolism Concept is similar to Metabolizable energy

23. FeedRumenIntestine Digestion Microbes Undegraded feed Metabolizable protein Protein Metabolism in the Rumen Less Extensively Degraded Protein

24. FeedRumenIntestine Digestion Microbes Undegraded feed Metabolizable protein Protein Metabolism in the Rumen Extensively Degraded Protein NH 3

25. Metabolizable Protein Supply to Host Animal Metabolizable protein (MP): Microorganisms – Digestible proteins Undegraded feed proteins – Digestible proteins Microorganisms g/d = 0.13 (TDN intake, g/d) (0.8) (0.8) Microbes 80% true protein that is 80% digested Feed g/d = (Feed protein) (Portion undegraded) (0.8) Feed proteins 80% digested

26. Absorption of Amino Acids Amino acids and small peptides absorbed by active transport (specific for groups of AA) From intestinesPortal blood Transport of amino acids into cells is similar process From bloodCells Active transport, requires energy

27. Utilization of Absorbed Amino Acids Via portal vein to liver Used for synthesis of proteins in liver Metabolized (deaminated) - Used for energy – Carbon for glucose Escape the liver Carried by blood to body tissues Used for synthesis of tissue proteins, milk, fetal growth, wool Metabolized - Used for energy

28. Requirements for Absorbed Amino Acids Metabolizable Protein (MP) Protein (amino acid) requirements 1.Maintenance 2.Growth 3.Lactation 4.Pregnancy 5.Wool

29. Protein Metabolism Concept of Net Protein Net protein = protein gained in tissues, milk, or fetal growth = NP Metabolizable protein is used with less than 100% efficiency Net protein = (MP - Metabolic loss) As a quantity, net protein is less than metabolizable protein

30. Net Protein Required for Production Amino AcidsProteins Milk kg/d = (Milk yield, kg/d) (% protein in milk) Growth g/d = SWG (268 - (29.4 (RE/SWG))) SWG = Shrunk weight gain, kg/d RE = Retained energy, Mcal/d RE obtained from net energy equations.

31. Protein Metabolism Metabolic Loss Protein synthesis and metabolism of amino acids draw from the same pool Proteins Amino acids Metabolism Metabolic loss results from continuous catabolism from amino acid pools Continuous turnover of tissue proteins adds to amino acid pools in tissues

32. Amino Acid (MP) Requirements Maintenance (three fractions) Protein required to support zero gain or production 1. Metabolism Metabolized Urine Milk Amino acids Feces Wool (Synthesis) GIT Scurf (Degradation) Pregnancy Tissue proteins = Endogenous urinary N 2. Proteins lost from body surface (hair, skin, secretions) = Scurf proteins 3. Proteins lost from undigested digestive secretions and fecal bacteria = Metabolic fecal N

33. Papers for Lab 4/8/10 doi:10.2527/jas.2009-2218 – “Effects of partial ruminal defaunation on urea- nitrogen recycling, nitrogen metabolism, and microbial nitrogen supply in growing lambs fed low or high dietary crude protein concentrations” doi: 10.2527/jas.2005-614 –“Effects of ractopamine and protein source on growth performance and carcass characteristics of feedlot heifers”

34. Figure Assignments Urea-N recycling paper –Table 1-group discussion –Table 2-Jessica A. –Table 3-Kenny B. –Table 4-JJ G. –Figure 1 and 2-Dan K. –Table 5-Kim M. –Table 6-Amir N.

35. Figure Assignments Ractopamine paper –Table 1-Jose N. –Table 2-Danielle P. –Table 3-Erin R. –Table 4-Nathan U.