Slide 1: Recent Developments in Performance Horse Health and Nutrition
Recent Developments in Performance Horse Health and Nutrition Joe D. Pagan, Ph.D. Kentucky Equine Research, Inc.
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Slide 3: Nutrition of the Performance Horse
Nutrition of the Performance Horse Energy is the nutritional factor most influenced by training and work
Slide 4: Energy Generation by Horse Muscle During Exercise
Energy Generation by Horse Muscle During Exercise Adenosine triphosphate ATP generated from: • blood glucose • muscle glycogen • fat • protein
Slide 5: Dietary Energy Sources
Dietary Energy Sources §Plant Fiber §Non-structural carbohydrate (NSC) • Starch • Sugar §Fat §Protein
Slide 6: Dietary Energy Sources
Dietary Energy Sources §Plant Fiber §Non-structural carbohydrate (NSC) • Starch • Sugar §Fat §Protein
Slide 7: Horse's Natural Feed is Forage
Horse’s Natural Feed is Forage Horse evolved as wandering herbivores with voluminous hindguts adapted to process large quantities of high fiber forage.
Slide 8: Domestication
Domestication • The energy requirements of working horses could not be met by forage alone • The deficit was met by feeding cereal grains
Slide 9: Dietary Energy Sources
Dietary Energy Sources §Plant Fiber §Non-structural carbohydrate (NSC) • Starch • Sugar §Fat §Protein
Slide 10: Glucose is a major substrate for energy generation
Glucose is a major substrate for energy generation
Slide 11: Aerobic and anaerobic energy generation
ATP Heart Lungs 36 ATP O2 CO2 2 ATP AEROBICALLY (WITH OXYGEN) ANAEROBICALLY (WITHOUT OXYGEN) GLUCOSE LACTIC ACID
Slide 12: Non-structural Carbohydrates (NSC)
Non-structural Carbohydrates (NSC) • Starch • Water Soluble Carbohydrate (WSS)
Slide 13: Horses are poor starch digesters
Horses are poor starch digesters • Rapid rate of passage • Low amylase production • Oats>Barley>Corn • Up to 70% of corn starch may escape prececal digestion
Slide 14: When large grain meals are fed to horses
When large grain meals are fed to horses • Starch may escape digestion in the small intestine • Rapidly fermented in the cecum and colon • Volatile fatty acid (VFA) and lactic acid production increases • A significant decrease in pH • Hindgut Acidosis (HGA)
Slide 15: Effects of Hindgut Acidosis
Effects of Hindgut Acidosis • Causes irritation or damage to the intestinal mucosa • Increases the permeability of the large intestinal mucosa to toxins and larger molecules • Implicated in the development of equine laminitis
Slide 16: Effects of Hindgut Acidosis
Effects of Hindgut Acidosis • Anorexia • Colic • Stereotypical behaviors such as wood chewing and stall weaving
Slide 17: Rumen acidosis
Rumen acidosis • Common problem in dairy NaHCO3 cattle fed high grain diets • Sodium bicarbonate is often added to a cows ration as a buffer • Attenuates drop in rumen pH that decreases feed intake and milk production
Slide 18: How do you deliver NaHCO3 to the hindgut?
How do you deliver NaHCO3 to the hindgut? NaHCO3 • Needs to survive acidic environment in stomach • Needs to survive digestive enzymes in small intestine • Needs to dissociate in the cecum and colon to provide bicarbonate to hindgut
Slide 19: EquiShure
EquiShureÒ • In partnership with Balchem, Kentucky Equine Research, Inc. developed an encapsulated sodium bicarbonate that survives transit through the stomach and small intestine of the horse
Slide 20: Feeding Protected Sodium Bicarbonate Attenuates Hindgut Acidosis in Horses Fed a High Grain Ration
Feeding Protected Sodium Bicarbonate Attenuates Hindgut Acidosis in Horses Fed a High Grain Ration Joe D. Pagan, PhD, T.J. Lawrence, MS and L.A. Lawrence, Ph.D. Kentucky Equine Research, Versailles, Kentucky 40383 Presented at 2007 AAEP Convention
Slide 21: Fecal pH
6.7 Control 6.6 EquiShure 6.5 Fecal pH 6.4 6.3 6.2 6.1 * *Sig time effect P<0.05 6.0 5.9 0 2 4 6 8 Time post feeding (hrs)
Slide 22: Fecal lactate mmol/l
3.7 * Control Fecal lactate mmol/l 3.2 EquiShure 2.7 2.2 1.7 *sig diff between 1.2 groups (P<0.05) 0.7 0.2 0 2 4 6 Time (hrs)
Slide 23: Dietary Energy Sources
Dietary Energy Sources §Plant Fiber §Non-structural carbohydrate (NSC) • Starch • Sugar §Fat §Protein
Slide 24: Muscle cell, mitochondria and muscle contraction
MUSCLE CELL MITOCHONDRIA MUSCLE CONTRACTION
Slide 25: 2nd International Conference on Equine Exercise Physiology, 1986
2nd International Conference on Equine Exercise Physiology, 1986
Slide 26: Effects of fat adaptation on glucose kinetics and substrate oxidation during low-intensity exercise
Effects of fat adaptation on glucose kinetics and substrate oxidation during low-intensity exercise J. D. PAGAN, R. J. GEOR, P. A. HARRIS, K. HOEKSTRA, S. GARDNER, C. HUDSON, A. PRINCE Volume 34, Issue S34, pages 33–38, September 2002
Slide 27: Digestible Energy Contribution
Digestible Energy Contribution CONTROL FAT
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Slide 29: Glucose Kinetics
Glucose Kinetics Liver glucose production (Ra) Glucose utilization (Rd) 35 30 Glucose Rd (µmol/kg/min) 30 Glucose Ra (µmol/kg/min) 25 25 20 20 15 15 10 10 5 5 0 0 -15 0 15 30 45 60 75 90 -15 0 15 30 45 60 75 90 Time (min) Time (min) CON 0 weeks FAT 0 weeks CON 5 weeks FAT 5 weeks
Slide 30: Respiratory Exchange Ratio
Respiratory Exchange Ratio Respiratory Exchange Ratio 1.00 Fat 0 0.95 Fat 10 0.90 0.85 * 0.80 * ** * * 0.75 0 10 20 30 40 50 60 70 80 90 Time (min)
Slide 31: Dietary Fat and Exercise Metabolism
Dietary Fat and Exercise Metabolism • After 5 weeks on a fat-supplemented diet (30% fat calories) horses used less carbohydrate and more fat during prolonged exercise • Implications –Sparing of carbohydrate desirable for horses undertaking endurance-type exercise
Slide 32: Horse muscle uses glycogen as its preferred fuel during strenuous exercise
Horse muscle uses glycogen as its preferred fuel during strenuous exercise
Slide 33: Effect of non-structural carbohydrate, fat and fiber intake on glycogen repletion following intense exercise
Effect of non-structural carbohydrate, fat and fiber intake on glycogen repletion following intense exercise 1Mesquita VS, 2Pagan JD, 3Valberg SJ, 2Waldridge BM, 2Whitehouse C. 1Universidade Estadual Paulista, Jaboticabal, São Paulo, Brazil; 2Kentucky Equine Research, Versailles, KY, United States;
Slide 34: Digestible Energy Contribution (% of Total DE)
DIGESTIBLE ENERGY CONTRIBUTION (% OF TOTAL DE) High NSC PROTEIN Moderate NSC Low NSC 15% PROTEIN PROTEIN 16% FAT FAT 15% FAT 11% 15% 23% CHO FIBER CHO FIBER FIBER CHO 18% 29% 37% 33% 43% 45%
Slide 35: Materials and Methods: Glycogen Depletion
Materials and Methods: Glycogen Depletion • 3 days of intense treadmill exercise 3° slope 1. Incremental maximal exercise test 2. Interval workout 3. Interval work out Sprint Interval Exercise Rest Fed Treatment 3-Day Glycogen Glycogen Repletion Diet Depletion Pre Post 24h 48h 72h 3wks training on d
Slide 36: Muscle glycogen (%)
muscle glycogen (%) 110+ 100+ depletion ¥ repletion 8 days post depletion x v -® CARBOHYDRATE -* FAT AND FIBER
Slide 37: Transcriptomic analysis of pathways limiting glycogen repletion in the horse
Transcriptomic analysis of pathways limiting glycogen repletion in the horse Deborah Vélez-Irizarry1, J Pagan2, V Mesquita2, SJ Valberg1 1McPhail Equine Performance Center, Department of Large Animal Clinical Sciences Michigan State University 2 Kentucky Equine Research, Versailles KY
Slide 38: Glucose Transport
Glucose Transport Glycogen synthesis • Rate limiting step - glucose transport into muscle cell GLUT1 GLUT4
Slide 39: Glucose transport
Glucose transport Glycogen Resynthesis Rate 4h Post Exercise Physiological in vitro insulin with CHO Administration stimulation 40 • Humans: 80% ↑GLUT4 30 translocation mmol kb btw/h 20 • Horses: 15% ↑GLUT4 10 translocation 0 Horse Human Lacombe et al Int Schol Res Articles 2014 Waller et al Equine
Slide 40: Glucose transporters
Glucose transporters Aerobic exercise Resistance training • GLUT1 • GLUT1 • GLUT3 • GLUT3 • GLUT4 • GLUT4 • GLUT5 • GLUT5 • GLUT6 • GLUT6 • GLUT8 • GLUT8 • GLUT10 • GLUT10 • GLUT11 • GLUT11 • GLUT12 • GLUT12
Slide 41: Methods: Quantify Gene Expression
Methods: Quantify Gene Expression Quality Filter: Tophat2: RNA-Seq: Adapter sequences Map reads to reference 2 x 150bp, pared-end Reads Reads Low quality reads EquCab3.0 100% 74% Paired Reads Mapped Reads Reads 59% 43 million 79% All expressed GLUT HTSeq: Differential Gene Expression Analysis: Quant
Slide 42: Glucose Transporter Gene Expression
Glucose Transporter Gene Expression HIGH NSC LOW NSC HIGH FAT *GLUT5 Compared to pre-exercise Low gene expression 24 hrs post depletion in high fat group
Slide 43: Conclusions
Conclusions • Horses lack signaling mechanisms that activate GLUT4 transcription following glycogen depletion • Lag in glycogen resynthesis on Low NSC-High Fat corresponds to lag in GLUT6 and GLUT10 expression • Difficult to replete muscle glycogen without adequate carbohydrate in the diet
Slide 44: Fat
FAT Crude Fat (mostly triglycerides) • Saturated Fatty Acids (SFA) • Monounsaturated Fatty Acid (MUFA) • Polyunsaturated Fatty Acids (PUFA) -Omega-3 Total -Omega-6 Total
Slide 45: Saturated and Unsaturated Fatty Acid
Saturated Fatty Acid ree a Ser tt th H H H H H H H H H H Unsaturated Fatty Acid O H H H HH toto | | tJ 4 HO—C—C—C—C—C— / va 1 | dt 4 \Z H H H H ES Ps o kp 4% Tan * Y Xf ? rN 4 %
Slide 46: Stearic, oleic and linoleic acid
Stearic acid C,,H,,O0, 18° °36 A saturated fat Oleic acid C,,H,,0, 18 34 A monounsaturated fat Linoleic acid C,,H,,0, 18° “32 A polyunsaturated fat H, H, H, H, 9? Cc c c ccc cc C-OH Ze Re Re I Oe ee NO ie H, H, H, H, 2 H,C G cre ¢ oe cc €& Ee H, H, H, H, H, H, H, 4H, double bond H, H H MIN OH OH OH
Slide 47: Polyunsaturated fatty acids
Polyunsaturated fatty acids Omega 3 and Omega 6
Slide 48: Polyunsaturated Fatty Acids (PUFAs) can be further divided by chain length
Polyunsaturated Fatty Acids (PUFAs) can be further divided by chain length • Short-chain polyunsaturated fatty acids (SC-PUFAs) have 18 or fewer carbon atoms • ALA, SDA and LA, GLA • Long-chain polyunsaturated fatty acids (LC- PUFAs) have ≥20 carbons in their length • EPA, DHA and DGLA, AA
Slide 49: Omega 3:6 ratio in pasture
Omega 3:6 ratio in pasture Pasture ALA:LA ratio 6 Pasture contains over 3 times 3.6 3.2 ALA:LA ratio 4 more omega 3 2 than omega 6 fatty acids 0 e e ur ur st st Pa Pa y a ck id or tu Fl en K
Slide 50: Soy Oil and Corn Oil
Soy Oil Corn Oil Fatty acid sources (% crude fat) Fatty acid sources (% crude fat) LA (18:2n-6) LA (18:2n-6)
Slide 51: Flaxseed oil and Fish oil
Flaxseed oil Fish oil (EO-3) Fatty acid sources (% crude fat) Fatty acid sources (% crude fat) (20:5n-3 + 22:6n-3) ALA (18:3n-3)
Slide 52: Omega-3 and Omega-6 pathways
OMEGA 6 OMEGA 3 @ Plant Sources = aaa sie — A 6-desaturase ——> | +<——— Elongase--5 —-——————> |—— A 5-desaturase ———————> | Elongase-2 I > ¥ SC-PUFA LC-PUFA Equine =e Research . ia
Slide 53: Omega-6 and omega-3 pathways
EXCESS LA INTAKE Omega 6 pathway Vegetable oils Omega 3 pathway Inflammatory LA (18:2n-6) ALA (18:3n-3) Novel oil Anti-inflammatory GLA (18:3n-6) DGLA (20:3n-6) X Anti-inflammatory EPA (20:4n-3) Fish oil Inflammatory Inflammatory AA (20:4n-6) DHA (22:6n-3) resolution
Slide 54: Kentucky Equine Research Performance Center, Ocala Florida
Kentucky Equine Research Performance Center Ocala Florida
Slide 55: Thoroughbreds
13 Thoroughbreds Three 90-day periods • PRE supplementation period • Two treatment periods • LC-PUFA or SC-PUFA • Switch-back design
Slide 56: Supplemental Omega Intake
Supplemental Omega 3 Intake 11 10 9 daily intake (g/d) 8 ALA C18:3n3 7 3.5 9.9 6 EPA C20:5n3 5 DPA C22:5n3 4 3 4.3 ALA DHA C22:6n3 2 1 0 LC-PUFA SC-PUFA OIL SOURCE
Slide 57: RBC EPA
RBC EPA 1.0 LC-PUFA *** 0.8 *** 0.65 SC-PUFA RBC EPA % 0.54 0.6 *** 0.42 0.4 0.2 0.10 0.08 0.09 0.07 0.09 0.0 0 1 2 3 0 1 2 3 month of supplementation *** different from month 0 (p<.001)
Slide 58: RBC DHA
RBC DHA 1.0 **** 0.83 LC-PUFA 0.8 *** SC-PUFA 0.66 **** RBC DHA % 0.56 0.6 0.4 0.2 0.10 0.12 0.09 0.08 0.07 0.0 0 1 2 3 0 1 2 3 month of supplementation *** different from month 0 (p<.001) **** different from month 0 (p<.0001)
Slide 59: Supplemental Omega Intake
Supplemental Omega 6 Intake 10 9 8 AA C20:4n6 daily intake (g/d) 7 6 GLA C18:3n6 5 5.3 8.8 LA C18:2n6 4 3 2 1 2.4 0 LC-PUFA SC-PUFA OIL SOURCE
Slide 60: RBC GLA and DGLA
RBC GLA RBC DGLA 0.5 0.8 *** **** LC-PUFA LC-PUFA *** 0.4 *** 0.57 0.4 0.3 SC-PUFA 0.50 SC-PUFA **** 0.6 *** RBC DGLA % RBC GLA % 0.3 0.44 0.3 0.4 0.29 0.2 0.25 0.25 0.1 0.1 0.1 0.1 0.24 0.25 0.1 0.1 0.2 0.0 0.0 0 1 2 3 0 1 2 3 0 1 2 3 0 1 2 3 month of supplementation month of supplementation *** di
Slide 61: Red Blood Cell LC-PUFAs after 90 days supplementation
Red Blood Cell LC-PUFAs 90 days supplementation 2.5 inflammatory 0.9:1 AA C20:4n6 2.0 3:1 3.3:1 1.5 anti-inflammatory % 1.0 DGLA C20:3n6 EPA C20:5n3 0.5 DHA C22:6n3 0.0 E E -P A FA LC UFA FA PR PR SC UF U U -P -P -P SC LC OIL SOURCE
Slide 62: Standardized exercise test (SET)
Standardized exercise test (SET) • 1800 m (9 f) trot • 1200 m (6 f) canter • 600 m (3 f) fast gallop • 1000 (5 f) m warm-down
Slide 63: Response to strenuous exercise
Response to strenuous exercise LC-PUFA supplementation resulted in: ü Reduced heart rate during breeze ü Enhanced mitochondrial biogenesis ü Reduced red blood cells in lungs ü Reduced eosinophils in lungs ü Increased anti-inflammatory cytokines (IL-10) ü Decreased inflammatory cytokines (IL-8) ü Red
Slide 64: Conclusions
Conclusions • LC-PUFA (DGLA-EPA-DHA) have beneficial effects on • Inflammation • Joints • Airways • EIPH • Gastric ulcers • KER has developed a new unique source of LC-PUFA for horses
Slide 65: Dietary Energy Sources
Dietary Energy Sources §Plant Fiber §Non-structural carbohydrate (NSC) • Starch • Sugar §Fat §Protein
Slide 66: A comparison of grain, oil and beet pulp as energy sources for the exercised horse
A comparison of grain, oil and beet pulp as energy sources for the exercised horse KATHLEEN G. CRANDELL, J. D. PAGAN, PAT HARRIS, S. E. DUREN Volume 31, Issue S30, pages 485–489, July 1999
Slide 67: Beet pulp as an energy source
Beet pulp as an energy source Beet pulp contains “super fiber” • Fiber is 75-80% digestible • Digestible Energy almost as high as oats Digestible Energy Distribution
Slide 68: Dietary Energy Sources
Dietary Energy Sources §Plant Fiber §Non-structural carbohydrate (NSC) • Starch • Sugar §Fat §Protein
Slide 69: Muscle Development
Muscle Development Skeletal muscle mass (SMM) = Muscle protein synthesis (MPS) – Muscle protein breakdown (MPB)
Slide 70: Muscle protein synthesis pathway
Extracellular space |! AMPK PKB/Akt Pk ae Intracellular space p7osek 4E-BP1 elF4G + muscle protein synthesis
Slide 71: What form of energy should we supply in performance horse feeds?
What form of energy should we supply in performance horse feeds?
Slide 72: Muscle Glycogen Utilization as Function of Speed: Endurance
Muscle Glycogen Utilization as Function of Speed ENDURANCE
Slide 73: Muscle Glycogen Utilization as Function of Speed: Eventing
Muscle Glycogen Utilization as Function of Speed eventing
Slide 74: Muscle Glycogen Utilization as Function of Speed: Racing
Muscle Glycogen Utilization as Function of Speed racing
Slide 75: Higher NSC diets needed for strenuous exercise (racing)
Higher non-structural carbohydrate (NSC) diets needed for strenuous exercise (racing) • Hindgut acidosis a concern • Attenuate with § Grain processing § Smaller meal size § Encapsulated Buffers (EquiShure) § Pre, Pro, and Post biotics may also help (more research needed)
Slide 76: Higher fat diets appropriate for lower intensity exercise
Higher fat diets appropriate for lower intensity exercise (sport and endurance horses) • Omega 6:3 PUFA imbalances an issue § Vegetable SC-PUFA (ALA) (flax oil) are not efficiently elongated to LC-PUFA § LC-PUFA supplementation needed to raise DGLA, EPA and DHA
Slide 77: Highly fermentable fiber sources (super fibers) warranted for all performance horse feeds
Highly fermentable fiber sources (super fibers) warranted for all performance horse feeds • Beet pulp • Soy hulls Amount added depends on desired balance of energy sources in feed
Slide 78: Quality protein important for performance horse feeds
Quality protein important for performance horse feeds • Stimulate muscle synthesis • BCAA • Substrate for muscle synthesis • Lysine • Methionine • Threonine
Slide 79: Energy Partitioning in Performance Feeds
Energy Partitioning in Performance Feeds % total Digestible Energy (DE) FIBER PROTEIN PROTEIN FIBER FAT FAT NSC NSC Racing Feed Sport Feed
Slide 80: Recent Developments in Performance Horse Health and Nutrition
Recent Developments in Performance Horse Health and Nutrition Joe D. Pagan, Ph.D. Kentucky Equine Research, Inc.