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Variation in the Susceptibility to Ruminal Acidosis G.B. Penner Department of Animal and Poultry Science University of Saskatchewan Email: [email protected] 1 Prevalence rates of SARA in dairy cows • 12-30% prevalence across lactation – Diagnosed using ruminocentesis 35.0% 30.0% n = 662 25.0% 20.0% 15.0% 10.0% 5.0% 0.0% 2 Krause and Oetzel, 2006 Acute ruminal acidosis pH 5.0 pH 5.5 Epithelial damage Decreased fibre digestibility Changes in rumen bacteria Increased VFA pH 6.0 Increased lactic acid SARA Healthy rumen conditions Consequences of ruminal acidosis 3 How much variation do we see? 4 Variation in the Severity of Ruminal Acidosis Duration below pH 5.8, h/d 24 Minimum Average Maximum 20 16 12 8 4 0 Day 0-5 Day 17-19 Day 37-39 Day relative to parturition Day 58-60 Penner et al., 2007 5 Variation in ruminal acidosis around parturition Fairfield et al., 20076 Is the Response to acidosis consistent? Salivary buffer Absorption Passage 7 20 18 16 14 12 10 8 6 4 2 0 10 9 8 7 6 5 4 3 2 1 0 DMI Duration <5.8 -5 5 17 37 Day relative to parturition Duration pH < 5.8, h/d DMI, kg/d Severity is not entirely related to DMI 58 8 Penner et al., 2007 Resistant vs. susceptible heifers Control Resistant Susceptible 3.0 6.9 6.7 2.0 6.5 1.5 6.3 1.0 6.1 0.5 5.9 0.0 5.7 -0.5 100% Hay 60% Hay 40% Hay 25% Hay Diet 15% Hay 8% Hay Mean rumen pH Starch intake, kg/d 2.5 5.5 9 Penner et al., unpublished Subacute Ruminal Acidosis Acid production Acid removal •Neutralization •Clearance 10 Acid Production in the Rumen Many glucose molecules ++ 2 acetate+ 2H 2H 2 propionate + + + 2H 2H ACID 1 Glucose pH 2 Pyruvate 1 butyrate + 1H 1H++ 11 Acid Removal from the Rumen bicarbonate blood Saliva H-SCFA SCFAAcid pH 12 Physiological for variation Variation in in Physiological Causes causes for the Susceptibility susceptibility to to Ruminal ruminal acidosis the Acidosis Acid removal from the rumen blood Passage Salivary 15.5% buffer Bicarbonate 28.0% Absorption Other 3.6% Passage Absorption 52.9% Allen, 1997 13 Regulating ruminal pH Acid Production •Salivary buffer Acid •Absorption Removal •Passage 14 Relationship between saliva production and ruminal pH Duration pH <5.8, h/d 25.00 Duration pH <5.8 = 0.07 × saliva production – 4.7 P < 0.001 20.00 15.00 10.00 5.00 0.00 150.00 200.00 250.00 300.00 350.00 400.00 Total saliva production, L/d Bowman et al., 2003; Yang et al.,152003 Regulating ruminal pH Acid Production •Salivary buffer Acid •Absorption Removal •Passage 16 Rumen pH ~5.0 to 7.0 Intracellular pH ~7.4 % in dissociated state Mechanisms: passive diffusion 100 90 80 70 60 50 40 30 20 10 0 1 2 3 4 5 6 7 8 9 10 pH H-SCFA H+ SCFAHCO3H2O + CO2 Carbonic anhydrase Ruminal contents Blood 17 SCFA absorption and H+ removal 2K+ Na+ Na+/K+ ATPase 3Na+ H+ H+ KetonesLactate- H-SCFA H+ Metabolism SCFAHCO3- SCFA- Apical surface pH > 7.4 H2O + CO2 Ruminal contents Leonhard-Marek et al., 2006 Blood 18 SCFA absorption and H+ removal Na+ H+ Na+ H+ H+ 2K+ 3Na+ KetonesLactate- H-SCFA H+ Metabolism SCFAHCO3- SCFA- H2O + CO2 Ruminal contents Blood 19 Mechanisms of SCFA absorption 20 Acetate uptake, nmol/(mg protein × min) Mechanisms: protein mediated 5.0 4.5 4.0 P < 0.001 a 3.5 3.0 2.5 b 2.0 1.5 b 1.0 0.5 0.0 Bicarbonate buffer Bicarbonate-free buffer Bicarbonate free buffer + nitrate 21 Penner et al., 2009 Acetate vs. butyrate Protein mediated 40-72% Uptake, nmol/(mg protein × min) 4.5 12.0 Passive diffusion 31-72% 4.0 10.0 3.5 3.0 2.5 Passive diffusion 28 – 60% 8.0 6.0 2.0 1.5 Protein mediated 28-69% 4.0 1.0 2.0 0.5 0.0 0.0 Bicarbonate Bicarbonate Bicarbonate dependent independent independent nitrate nitrate sensitve insensitive Bicarbonate dependent Bicarbonate independent nitrate sensitve Bicarbonate independent nitrate insensitive 22 Penner et al., 2009 SCFA absorption and H+ removal Na+ Aschenbach et al., 2010 H+ Na+ H+ H+ 2K+ 3Na+ H+ KetonesLactate- H-SCFA H+ Metabolism SCFAHCO3- HCO3- SCFAHCO3- H2O + CO2 Ruminal contents Blood Na+ HCO3- 23 SCFA absorption and ruminal pH • Appearance of bicarbonate in luminal buffer Buffer pH (Ash and Dobson, 1963; Gäbel et al., 1991) 9 8 7 6 5 4 3 2 1 0 Initial 1 Final 2 3 Buffer 4 Dijkstra et al., 1993 24 Variation in ruminal pH 7.0 Duration pH < 5.8, h/180 min 180 6.2 5.8 5.4 a P < 0.05 160 140 120 100 b 80 Control 60 40 Resistant c 20 Susceptible 0 Control Resistant Susceptible 0 -360 -720 -1080 -1440 -1800 -2160 -2520 5.0 -2880 Ruminal pH 6.6 Time relative to the challenge, min Penner et al., 2009 25 Uptake, nmol/cm2/h SCFA absorption explains variation in ruminal pH 16 Control 14 Resistant 12 Susceptible P <0.05 10 b b a 8 6 a b b 4 2 0 Acetate Butyrate 26 Mechanism of acetate uptake and ruminal pH Penner et al., 2009 27 Mechanism of butyrate uptake and ruminal pH •Non-responders also had higher serum butyrate Penner et al., 2009 28 SCFA absorption and ruminal pH Resende Júnior et al., 2006 r2 = 0.43 P <0.001 r2 = 0.45 P <0.001 Gene Acyl-CoA synthetase HMGCL LDHb PC r2 = 0.17 P <0.001 Pathway R P value Activation of SCFA -0.92 0.01 Ketogenesis -0.75 0.08 Lactate metabolism -0.84 0.03 Pyruvate metabolism 0.88 0.02 Penner et al., 2009 29 Take home messages • Considerable variation among cows for the susceptibility to ruminal acidosis • Variation in susceptibility is related to differences in the total absorption and pathway of absorption for SCFA • Future research is required to determine the molecular regulation for acidosis susceptibility 30 Thank You! 31