Effect of supplemental vitamin C on meat quality of cattle fed varying concentrations of dietary sulfur

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Effect of supplemental vitamin C on meat quality of cattle fed varying concentrations of dietary sulfur S.L. Hansen*, S.M. Lonergan, and D.J. Pogge Department of Animal Science Iowa State University

Introduction Distillers grains are a nutritional and inexpensive alternative to corn Distillers grains can be high in sulfur (S) and fat Ranges include: 4-12% fat (Stock et al., 2000) S varies depending on plant Can be over 1% S

Introduction High dietary S results in: Negative impacts on feedlot performance and meat quality 0.6% S decreased ADG, HCW Sulfur toxicity (PEM) Oxidative stress

Introduction Acceptance of meat products by consumers primarily related to tenderness and flavor/marbling (Miller et al., 1995) Oxidative environment post mortem: Oxidation of protease µ-calpain can limit the extent of muscle proteolysis = toughness Oxidation of lipids = off flavors = shelf-life

Oxidation Slows Proteolysis and Tenderization C 17 E 18 C 19 E 20 Non Non Non Non Ox Ox Ox Ox Ox Ox Ox Ox Intact Troponin-T { Troponin-T Degradation Product Kg Shear 4.2 2.7 3.9 2.9 3.0 2.6 3.4 2.7 Force Rowe, L.J., K.R. Maddock, S.M. Lonergan, E. Huff-Lonergan. 2004. J. Anim. Sci. 82:3254-3266.

Oxidation Influences Autolysis of µ-calpain Day 3 Post-Irradiation Irr NI Irr NI Irr NI Irr NI Supernatant Rowe, L.J., K.R. Maddock, S.M. Lonergan, E. Huff-Lonergan. 2004. J. Anim. Sci. 82:3254-3266

Effect of oxidation on progression of autolysis of µ-calpain Ox= pre exposure to 200 µm H2O2; Reduced = pre exposure to 0.1 % MCE Lametsch, Huff-Lonergan and Lonergan. BBA Proteins and Proteomics, 1784:1215

Vitamin C Research Cattle synthesize ascorbate (vitamin C) in the liver Roles in the body include: Antioxidant function (redox reactions) Regeneration of glutathione and vitamin E Synthesis of collagen and carnitine Enzyme cofactor

Vitamin C Research In vivo Japanese long fed cattle (Takahashi et al., 1999; Ohashi et al., 2000) Depletion of plasma vitamin C during finishing period Vitamin C added late in the finishing period increased marbling scores Jugular infusions of vitamin C improved color and lipid stability Cell culture work Vitamin C enhanced adipocyte differentiation (Kawachi, 2006)

Study Objectives Determine the effects of supplemental vitamin C in diets containing varying concentrations of dietary S on: 1. Performance of cattle 2. Carcass characteristics of cattle 3. Meat quality (protein degradation and oxidation)

Materials and Methods 120 steers, 5 pens/trt with 4 steers/pen 6 treatments Low Sulfur, 0.2% S No Vitamin C + Vitamin C Medium Sulfur, 0.4% S No Vitamin C + Vitamin C High Sulfur, 0.6% S No Vitamin C + Vitamin C Rumen-protected vitamin C source included at 10g of vitamin C/head/day

Diet composition and analysis Item Low S Medium S High S Corn 69.7 48.0 48.0 Corn dried distillers grains 18.0 40.0 38.9 Chopped hay 9.0 9.0 9.0 Limestone 2.0 2.0 2.0 Salt 0.31 0.31 0.31 Vitamin A premix 1 0.10 0.10 0.10 Trace mineral premix 2 0.035 0.035 0.035 Rumensin90 3 0.016 0.016 0.016 Sodium sulfate -- -- 1.11 Urea 0.8 -- -- S, % 0.18 0.32 0.56 CP, % 14.3 16.6 16.6 NEg, Mcal/kg DM 1.3 1.3 1.3 1 Vitamin A premix contains 4,400,000 IU/kg 2 Provided per kg of diet: 30 mg Zn as ZnSO 4 ; 20 mg Mn as MnSO 4 ; 0.5 mg I as Ca(IO 3 ) 2 (H 2 O); 0.1 mg Se as Na 2 SeO 3 ; 10 mg Cu as CuSO 4 ; and 0.1 mg Co as CoCO 3 3 Provided at 27 g/ton diet

Study Timeline 2010 2011

Measurements taken Live animal: BW, DMI, ADG, G:F, plasma vitamin C, plasma antioxidant capacity Carcass: HCW, back-fat, KPH, REA, YG, QG, marbling score Meat Quality Proximate analysis Troponin-T degradation-western Blotting µ-calpain autolysis-western Blotting Protein carbonylation-oxi-blots Iron analysis-icp-oes TBARS Fatty acid profile-gc-ms

Statistical Analysis Contrast Statements: A = Vitamin C vs. No Vitamin C B = Linear effect of Sulfur C = Vitamin C within High Sulfur D = Vitamin C within Sulfur E = Vitamin C within Corn Diet Significance denotations: (P 0.10); *(P 0.05); **(P < 0.01)

Results

Plasma Vitamin C Plasma Vitamin C, ug/l 1000 900 800 700 600 500 400 Low S Low S + VC Med S Med S + VC High S High S + VC Vitamin C effect (P = 0.04) Vitamin C within HS (P = 0.01) Vitamin C within S (P = 0.02) 300 d 0 d 28 d 90 d 143 Days on Study

Plasma Total Antioxidant 2.70 Low S Low S + Vitamin C Medium S Medium S + Vitamin C High S High S + Vitamin C Plasma Total Antioxidant, mm 2.50 2.30 2.10 1.90 1.70 1.50 1.30 1.10 0.90 Linear effect of S, d 90: P = 0.003 Linear effect of S, d 143: P = 0.10 0.70 d 0 d 90 d 143 Days on Study

Feedlot Performance Data Diet S Low Medium High Diet vitamin C - + - + - + SEM Sig 1,2 Pen (steers) 5 (20) 5 (20) 5 (20) 5 (20) 5 (19) 5 (20) Initial BW, kg 342 342 341 343 340 340 10.6 Intake DMI, kg/d 3 10.54 10.50 10.82 10.20 9.41 9.73 0.32 B** S intake, g/d 3 21.8 22.6 34.6 33.6 53.6 57.6 1.61 B**C VC intake, g/d 3-10.5-10.3-9.9 0.63 Live performance 4 Final BW, kg 576 568 575 573 545 556 12.4 B** ADG, kg/d 1.65 1.65 1.69 1.63 1.43 1.48 0.04 B** G:F 0.16 0.15 0.16 0.15 0.15 0.15 0.004 1 Contrast Statements: A= vitamin C vs. no vitamin C; B = Linear effect of sulfur; C = vitamin C within high sulfur; D = vitamin C within sulfur; E = vitamin C within corn diet 2 (P 0.10); *(P 0.05); **(P < 0.01) 3 Month and treatment month (P < 0.05) 4 Live performance values based on measured live BW with a 4% pencil shrink applied

Carcass Data Diet S Low Medium High Diet vitamin C - + - + - + SEM Sig 1,2 Pen (steers) 5 (20) 5 (19) 5 (20) 5 (19) 5 (18) 5 (20) HCW, kg 369 365 371 367 350 356 8.29 B** Dressing % 64.0 64.1 64.8 65.0 64.3 64.3 0.002 Fat, cm 1.48 1.28 1.28 1.24 1.00 1.24 0.06 B**C*D E* REA, cm 2 80.6 79.9 81.8 81.8 82.1 78.9 1.07 C* KPH 2.08 2.34 2.21 2.43 2.26 2.18 0.05 A**C*D*E** Calculated YG 3.45 3.28 3.26 3.20 2.90 3.16 0.14 B* Marbling score 3 511 443 474 438 398 470 13.4 B**C**E** QG 4 3.45 3.15 3.45 3.00 2.75 3.35 0.17 B C* 1 Contrast Statements: A= vitamin C vs. no vitamin C; B = Linear effect of sulfur; C = vitamin C within high sulfur; D = vitamin C within sulfur; E = vitamin C within corn diet 2 (P 0.10); *(P 0.05); **(P < 0.01) 3 Marbling Scores, Traces: 200, Slight: 300, Small: 400, Modest: 500, Moderate: 600 4 Quality Grade refers to the plant assigned QG

Proximate analysis Diet sulfur Low Medium High Diet vitamin C - + - + - + SEM Sig 1,2 Item Pen (steers) 5(13) 5(13) 5(15) 5(15) 5(14) 5(15) Moisture, % 71.9 72.2 72.1 72.4 73.2 73.0 0.29 B** Fat, % 5.40 5.03 5.48 5.05 3.56 4.29 0.38 B** Protein, % 22.3 22.1 21.5 22.0 22.3 21.9 0.10 C 1 Contrast Statements: A= vitamin C vs. no vitamin C; B = Linear effect of sulfur; C = vitamin C within high sulfur; D = vitamin C within sulfur; E = vitamin C within corn diet 2 (P 0.10); *(P 0.05); **(P < 0.01)

µ-calpain autolysis Percentage of Catalytic Subunit Intact 60 50 40 30 20 10 0 Linear effect of S (P = 0.03); vitamin C within high S (P < 0.09) Vitamin C (P = 0.09); vitamin C within high S (P < 0.05); vitamin C within C medium and high S (P = 0.03) Linear effect of S (P = 0.107) 80 kda 78 kda 76 kda Catalytic Subunits Day 2 Postmortem Low S Low S + vitamin C Medium S Medium S + vitamin C High S High S + vitamin C

Troponin-T degradation product Peak Height Ratio 1 0.9 0.8 0.7 0.6 0.5 0.4 0.3 0.2 0.1 0 Linear effect of S: P < 0.01 Day 2 Postmortem Low S Low S + vitamin C Medium S Medium S + vitamin C High S High S + vitamin C

Protein carbonylation 40000 35000 Linear effect of S: P < 0.04 Low S Peak Height 30000 25000 20000 15000 10000 5000 Low S + vitamin C Medium S Medium S + vitamin C High S High S + vitamin C 0 Day 2 Postmortem

TBARS and Iron analysis Diet S Low Medium High Diet vitamin C - + - + - + SEM Sig 1,2 Item Pen(steers) 5(18) 5(17) 5(19) 5(20) 5(18) 5(18) Fe, mg/kg 12.8 14.7 12.7 13.8 14.0 13.1 0.46 A C* Pen(steers) 5(13) 5(13) 5(12) 5(17) 5(12) 5(16) MDA 3, µm 0.29 0.33 0.29 0.24 0.27 0.33 0.014 A C*E** 1 Contrast Statements: A= vitamin C vs. no vitamin C; B = Linear effect of sulfur; C = vitamin C within high sulfur; D = vitamin C within sulfur; E = vitamin C within corn diet 2 (P 0.10); *(P 0.05); **(P < 0.01) 3 Malondialdehyde (lipid peroxidation product)

Summary Increasing dietary S Decreased the presence of the 76 kda subunit of the protease µ-calpain, troponin-t degradation, protein carbonylation, & percent fat in rib facings Increased: percent protein of rib facings Addition of vitamin C Increased marbling scores of high S cattle Increased fatness of cattle Increased 76 kda subunit of µ-calpain in all treatments, but specifically noted in high S Increased TBARS and Fe concentration in muscle

Conclusion and Implications Adding vitamin C to high S diets may increase both the quality grade and tenderness of the final meat product DDGS-based diets are often high in S, feeding vitamin C to cattle may allow producers enjoy the economic benefits of a DDGS-based diet while still maintaining the quality and tenderness expected by consumers

Acknowledgments Danielle Pogge, Ph.D. student Dr. Steven Lonergan, meat scientist Farm crew at ISU Beef Nutrition Farm ICPB and IBIC for funding RMC for the opportunity to speak with you!