- Email: [email protected]
Evaluation of supplemental vitamin A and E on 56-day growth performance, dietary net energy, and plasma retinol and tocopherol concentrations in Holstein steer calves
The Professional Animal Scientist 30 (2014):510–514; http://dx.doi.org/10.15232/pas.2014-01316 ©2014 American Registry of Professional Animal Scientists
Evaluation of supplemental vitamin A and E on 56-day
growth performance, dietary net energy, and plasma retinol and tocopherol concentrations in Holstein steer calves J. Salinas-Chavira,* A. A. Arrizon,† A. Barreras,† C. Z. Chen,‡ A. Plascencia,† and R. A. Zinn§ *Facultad de Medicina Veterinaria y Zootecnia, Universidad Autonoma de Tamaulipas, Cd. Victoria, Tam. 87000, México; †Instituto de Investigaciones en Ciencias Veterinarias, Universidad Autonoma de Baja California, Mexicali, Baja California 21100, México; ‡College of Animal Science, Jilin University, Changchun, China 130062; and §Department of Animal Science, University of California, Davis 95616
ABSTRACT Thirty Holsteins steers (initial weight 230 ± 1 kg) fed as calves were used in a 56-d experiment to evaluate the influence of vitamin A and E supplementation on growth performance, dietary energetics, and plasma retinol and tocopherol concentrations. Steers were fed a steamflaked, corn-based diet. Three treatments (TMT) were evaluated: TMT1 had no supplemental vitamins, TMT2 supplemented 30,000 IU/d of vitamin A as retinyl propionate plus 250 IU/d of vitamin E as all-rac α-tocopheryl acetate, and TMT3 supplemented 30,000 IU/d of vitamin A as retinyl palmitate plus 250 IU/d of vitamin E as d-α-tocopherol (RRR α-tocopherol). Treatment supple-
1 Corresponding author: razinn@ucdavis. edu
ments were freshly prepared and applied to feed once daily in the morning. Form of supplemental vitamins A and E did not affect cattle growth performance or dietary energetics. Vitamin supplementation increased final BW (3.7%, P = 0.02), ADG (10.7%, P = 0.02), and DMI (7.1%, P = 0.05). There were no treatment effects (P > 0.30) on gain efficiency or dietary net energy. There were no treatment effects on plasma retinol on d 28. However, by d 56, vitamin A supplementation increased (P = 0.01) plasma retinol by 56%. Form of supplemental vitamin A did not influence (P > 0.30) plasma retinol. Vitamin E supplementation increased (P < 0.01) plasma tocopherol. On d 28, plasma tocopherol tended to be greater (P = 0.12) with supplemental tocopheryl acetate than for supplemental tocopherol. By d 56 plasma tocopherol was 31% greater (P = 0.03) for all-rac α-tocopheryl acetate than for
RRR α-tocopherol supplemented steers. Vitamin supplementation may enhance DMI and corresponding ADG of Holstein steer calves fed a steam-flaked corn-based diet. Ester forms of vitamin A (retinyl propionate vs. retinyl palmitate) are comparable. Key words: feedlot, Holstein, vitamin A, vitamin E
INTRODUCTION Basal ingredients (grains and roughages) commonly used in growingfinishing diet formulations for feedlot cattle are characteristically deficient in supplying vitamin A and, possibly, vitamin E (NRC, 2000); therefore, dietary supplementation is a common practice. In addition to changes in growth performance, bioavailability of
511
Vitamins A and E on performance and plasma levels
supplemental vitamin A and E forms are often evaluated based on resulting changes in plasma concentrations, tissue concentrations, or both (Vagni et al., 2011). Response to vitamin A and E supplementation of feedlot cattle has been variable (Perry et al., 1968; Arnold et al., 1992; Bryant et al., 2010; Burken et al., 2012), due in part to cattle background and composition of the basal diet. There is limited information regarding the vitamin A and E status of Holstein steers fed as calves a conventional steam-flaked, corn-based, growing-finishing diet. The objective of the current experiment was evaluate vitamin A and E supplementation of Holstein steers fed as calves, comparing the combinations of retinyl propionate and all-rac α-tocopheryl acetate versus retinyl palmitate and RRR α-tocopherol.
MATERIALS AND METHODS All animal care, handling, and sample techniques followed protocols approved by the University of California, Davis, Animal Use and Care Committee.
Animals and Design Thirty Holstein steer calves (initial BW = 230 ± 1 kg) were used in a 56-d experiment to evaluate influence supplemental vitamin A and E on growth-performance, dietary NE, and plasma vitamin concentrations. Steers were blocked by initial BW and assigned within BW groupings to 15 pens, 2 steers/pen. Pens were 5.5 × 9.1 m with 27 m2 of shade and were equipped with automatic waterers and fence-line feed bunks (4.3-m long). Upon initiation, the steers were implanted with Revalor-S (Intervet Inc., Millsboro, DE). The current experiement used 3 dietary treatments (TMT) consisting of a steam-flaked, corn-based, growing-finishing diet (Table 1) and supplements: (TMT1) a control with no supplemental vitamins; (TMT2) 30,000 IU/d of vitamin A as retinyl propionate (Microvit A DLC 500 HF; Adisseo USA Inc.,
Table 1. Composition of experimental diets
Item
Basal diet, %
Ingredient composition (DM basis) Steam-flaked corn 62.96 Alfalfa hay 3.87 Sudangrass hay 7.74 2.67 Yellow grease Molasses cane 5.93 Distillers dried 14.36 0.58 Urea Limestone 1.45 Magnesium oxide 0.09 Monensin, mg/kg 30.00 Trace mineral salt1 0.35 Nutrient composition (DM basis)2 NE, Mcal/kg Maintenance 2.21 Gain 1.54 CP, % 13.91 Calcium, % 0.72 Phosphorus, % 0.35 Vitamin A, IU/kg 418 Vitamin E, IU/kg 25 Trace mineral salt contained: CoSO4, 0.068%; CuSO4, 1.04%; FeSO4, 3.57%; ZnO, 1.24%; MnSO4, 1.07%, KI 0.052%; and NaCl, 92.96%. 2 Based on tabular values for individual feed ingredients (NRC, 2000), with the exception of vitamin A which was estimated based on vitamin A concentration of feed ingredients reported by Pickworth et al. (2012). 1
Alpharetta, GA) plus 250 IU/d of vitamin E as all-rac α-tocopheryl acetate (Microvit E DLC 60 HF; Adisseo USA Inc.); and (TMT3) 30,000 IU/d of vitamin A as retinyl palmitate (Emcelle Vitamin A-Liquid; Stuart Products, Bedford, TX) plus 250 IU/d of vitamin E as RRR α-tocopherol (Emcelle Tocopherol; Stuart Products). Steers were provided ad libitum access to the experimental diets. Fresh feed was provided twice daily (0600 and 1400 h). Daily application of vitamin treatments was accomplished by dilution in 30 mL of water before top dressing onto feed provided at the 0600 h feeding.
At 28-d intervals, blood samples were collected via jugular puncture into heparinized tubes. The blood was immediately centrifuged at 1,070 × g for 10 min at 23°C. Plasma α-tocopherol and retinol analyses were conducted by the Veterinary Diagnostic Laboratory (College of Veterinary Medicine, Iowa State University, Ames) according to procedures described by Stahr (1991).
Estimation of dietary NE Daily energy gain (EG; Mcal/d) was calculated by the equation EG = ADG1.097 × 0.0557W0.75, where W is the mean shrunk BW (kg; NRC, 1984). Maintenance energy (EM) was calculated by the equation EM = 0.084W0.75 (Garrett, 1971). Dietary NEg was derived from NEm by the equation NEg = 0.877NEm – 0.41 (Zinn, 1987). Dry matter intake is related to energy requirements and dietary NEm according to the equation DMI = EM/NEm + EG/(0.877NEm − 0.41), and can be resolved for estimation of dietary NE by means of the quadratic formula
where x = NEm; a = −0.41EM; b = 0.877 EM + 0.41DMI + EG; and c = −0.877DMI (Zinn and Shen, 1998).
Statistical Design and Analysis For calculating steer performance, initial and final BW were reduced 4% to account for digestive tract fill. The experimental data were analyzed as a randomized complete block design according to the statistical model Yij = μ + Bi + Tj + εij (Hicks, 1973), where μ is the common experimental effect; Bi represents blocks (df = 4); Tj represents dietary treatment effect (df = 2), and εij represents the residual error (df = 8). Treatments effects were tested by means of orthogonal contrasts as (1) control versus supplemental vitamins and (2) TMT2 versus TMT3. Analysis was performed using
512
Salinas-Chavira et al.
Statistix-10 (Analytical Software, Tallahassee, FL).
vitamin A or E, or the combination of both. Perry et al. (1968) evaluated vitamin A and E supplementation of ground corn-based finishing diets fed to feedlot steer calves. Vitamins were fed either singly or in combination. Fed individually, vitamin A supplementation increased DMI and ADG, whereas vitamin E supplementation did not affect growth performance. Likewise, the combination of vitamin A and vitamin E supplementation did not affect growth performance beyond that observed for vitamin A supplementation. However, the effect of vitamin E supplementation of feedlot cattle on growth performance has not been consistent. Whereas numerous subsequent studies (Arnold et al., 1992; Zinn et al., 1996; Rivera et al., 2002; Carter et al., 2005; Montgomery et al., 2005) have likewise failed to show a growth performance response to supplemental vitamin E, in other instances vitamin E supplementation enhanced daily BW gain (Reddy et al., 1987; Pehrson et al., 1991; Burken et al., 2012). Daily BW gain response to vitamin E supplementation is apparent when plasma tocopherol
RESULTS AND DISCUSSION Effects of Treatments on Growth Performance and Dietary Energetics Treatment effects on cattle growthperformance and dietary NE are shown in Table 2. Form of supplemental vitamins A (retinyl propionate vs. retinyl palmitate) and E (RRR α-tocopherol vs all-rac α-tocopheryl acetate) did not affect cattle growth performance or dietary energetics. However, vitamin supplementation increased final BW (3.7%, P = 0.02), ADG (10.7%, P = 0.02), and DMI (7.1%, P = 0.05). The improvement in ADG is attributable to increased DMI, as there were no treatment effects (P > 0.30) on BW gain efficiency or dietary net energy. It is not possible in the present experiment to determine if beneficial treatment effects on growth performance were due to supplemental
levels are less than 1 μg/mL (Reddy et al., 1987; Pehrson et al., 1991). In the present experiment the minimum observed plasma tocopherol concentration of nonsupplemented steers was 1.4 μg/mL (Table 3). Kohlmeier and Burroughs (1970) observed that the critical level of plasma retinol below which supplementation enhanced ADG was 0.25 μg/mL. This observation is consistent with recent studies (Gorocica-Buenfil et al., 2007a,b; Gibb et al., 2011) where vitamin A supplementation did not enhance feedlot cattle growthperformance. Although, in an experiment reported by Bryant et al. (2010) where initial plasma retinol average 0.15 μg/mL, vitamin A supplementation also failed to enhance daily BW gain. However, in their study, plasma retinol of nonsupplemented steers increased with days on feed. In the present experiment, initial plasma retinol of nonsupplemented steers was 0.23 μg/mL and decreased with days on feed (Table 3), suggestive of marginal vitamin A stores in our Holstein steers fed as calves.
Table 2. Growth performance of Holstein calves that received 2 forms of vitamin A and E Treatment
Item Pen replications Live BW,4 kg Initial Final ADG, kg/d DMI, kg/d G:F Dietary NEm, Mcal/kg Dietary NEg, Mcal/kg Observed/expected NEm Observed/expected NEg
P-value
Control1
AE12
AE23
5
5
5
229 322 1.67 7.26 0.22 2.13 1.46 0.96 0.95
231 333 1.82 7.63 0.23 2.18 1.51 0.98 0.98
229 336 1.91 8.01 0.23 2.17 1.49 0.97 0.97
Control vs. supplemental vitamins
AE1 vs. AE2
SEM
0.53 0.02 0.02 0.05 0.32 0.50 0.50 0.50 0.50
0.310 0.541 0.330 0.216 0.990 0.825 0.825 0.825 0.825
1 3 0.05 0.18 0.01 0.05 0.04 0.02 0.03
Control = no supplemental vitamins. AE1 = vitamin A and E combination consisting of a daily dosage of 30,000 IU of vitamin A as retinyl propionate plus 250 IU of vitamin E as all-rac α-tocopheryl acetate. 3 AE2 = vitamin A and E combination consisting of a daily dosage of 30,000 IU of vitamin A as retinyl palmitate plus 250 IU of vitamin E as RRR α-tocopherol. 4 Body weights were reduced 4% to account for digestive tract fill. 1 2
513
Vitamins A and E on performance and plasma levels
Table 3. Treatments effects on plasma retinol and tocopherol concentrations of calves that received 2 forms of vitamin A and E Treatment
Item
Control1
Pen replications Plasma retinol, µg/mL 0d 28 d 56 d Plasma tocopherol, µg/mL 0d 28 d 56 d
P-value
AE12
AE23
5
5
5
0.23 0.21 0.16
0.21 0.23 0.26
2 2.04 1.36
1.81 3.84 3.30
0.23 0.25 0.25 2.17 3.31 2.52
Control vs. supplemental vitamins
0.59 0.15 0.01 0.97 0.00 0.00
TM2 vs. TMT3
0.36 0.38 0.78 0.24 0.12 0.03
SEM
0.01 0.01 0.02 0.18 0.20 0.18
Control = no supplemental vitamins. AE1 = vitamin A and E combination consisting of a daily dosage of 30,000 IU of vitamin A as retinyl propionate plus 250 IU of vitamin E as all-rac α-tocopheryl acetate . 3 AE2 = vitamin A and E combination consisting of a daily dosage of 30,000 IU of vitamin A as retinyl palmitate plus 250 IU of vitamin E as RRR α-tocopherol. 1 2
Effects of Treatments on Plasma Level of Vitamin A and E Treatment effects on plasma retinol and tocopherol concentrations are shown in Table 3. There were no effects of treatments on plasma retinol on d 28. However, by d 56, vitamin A supplementation increased (P = 0.01) plasma retinol by 56%. Form of supplemental vitamin A (retinyl propionate vs. retinyl palmitate) did not influence (P > 0.30) plasma retinol concentrations. Comparison of the 2 forms of vitamin A on plasma retinol concentrations has not been previously reported. Based on in vitro incubation research using ruminal fluid from steers fed a high-concentrate diet, Rode et al. (1990) observed that supplemental vitamin A (retinyl propionate) is extensively degraded by ruminal fermentation. After 12 h of incubation, 72% of retinol had been degraded, with degradation of retinol proceeding at a faster rate than hydrolysis of the retinyl ester. Based on an assumed ruminal dilution rate of 5%/h, they estimated that 33% of supplemental vitamin A reaches the small intestine of cattle
fed a high-concentrate diet. Assuming that 90% of vitamin A entering the small intestine is absorbed (Donoghue et al., 1983), then it is expected that approximately 30% of supplemental vitamin A was metabolizable. As expected (Pehrson et al., 1991; Arnold et al., 1992, 1993; Rivera et al., 2002), vitamin E supplementation increased (P < 0.01) plasma tocopherol concentration. On d 28, plasma tocopherol tended to be 16% greater (P = 0.12) with supplemental tocopheryl acetate than for supplemental tocopherol. By d 56, plasma tocopherol was 31% greater (P = 0.03) for all-rac α-tocopheryl acetate-supplemented steers than for RRR α-tocopherolsupplemented steers. This later result was unexpected. Based on standards of bioavailability (EFSA, 2010), the potency of RRR α-tocopherol is 49% greater than that of all-rac α-tocopheryl acetate. Accordingly, Hidiroglou et al. (1988) and Eicher et al. (1997) observed greater plasma tocopherol concentrations in cattle orally dosed with RRR α-tocopherol than with the all-rac α-tocopheryl acetate. In research using sheep (Hidiroglou et al., 1992) or dairy cattle (Weiss et al., 2009), dietary supplementation with
RRR α-tocopheryl acetate resulted in greater plasma tocopherol concentrations than supplementation with all-rac α-tocopheryl acetate. Comparative studies evaluating dietary supplementation of the free alcohol form of vitamin E have not been previously reported. However, the free alcohol form (RRR α-tocopherol) is more susceptible to oxidation than the ester forms (Vagni et al., 2011). Thus, although vitamin treatments were freshly applied to the feed daily, lower plasma tocopherol concentrations may reflect oxidative loss of the RRR α-tocopherol before consumption. More research is needed to evaluate differences in oxidative stability of vitamin E forms in growing-finishing diets for cattle.
IMPLICATIONS Vitamin supplementation may enhance DMI and corresponding ADG of Holstein steer calves fed a steamflaked, corn-based, growing-finishing diet. Ester forms of vitamin A (retinyl propionate vs. retinyl palmitate) are comparable. Based on plasma tocopherol levels, the basal nonsupplemented diet was not deficient in vitamin E.
514
Salinas-Chavira et al.
LITERATURE CITED Arnold, R. N., S. C. Arp, K. K. Scheller, S. N. Williams, and D. M. Schaefer. 1993. Tissue equilibration and subcellular distribution of vitamin E relative to myoglobin and lipid oxidation in displayed beef. J. Anim. Sci. 71:105–118. Arnold, R. N., K. K. Scheller, S. C. Arp, S. N. Williams, D. R. Buege, and D. M. Schaefer. 1992. Effect of long- or short-term feeding of α-tocopheryl acetate to Holstein and crossbred beef steers on performance, carcass characteristics, and beef color stability. J. Anim. Sci. 70:3055–3065. Bryant, T. C., J. J. Wagner, J. D. Tatum, M. L. Galyean, R. V. Anthony, and T. E. Engle. 2010. Effect of dietary supplemental vitamin A concentration on performance, carcass merit, serum metabolites, and lipogenic enzyme activity in yearling beef steers. J. Anim. Sci. 88:1463–1478. Burken, D. B., R. B. Hicks, D. L. VanOverbeke, G. G. Hilton, J. L. Wahrmund, B. P. Holland, C. R. Krehbiel, P. K. Camfield, and C. J. Richards. 2012. Vitamin E supplementation in beef finishing diets containing 35% wet distillers grains with solubles: Feedlot performance and carcass characteristics. J. Anim. Sci. 90:1349–1355. Carter, J. N., D. R. Gill, C. R. Krehbiel, A. W. Confer, R. A. Smith, D. L. Lalman, P. L. Claypool, and L. R. McDowell. 2005. Vitamin E supplementation of newly arrived feedlot calves. J. Anim. Sci. 83:1924–1932. Donoghue, S., W. J. Donawick, and D. S. Kronfeld. 1983. Transfer of vitamin A from intestine to plasma in lambs fed low and high intakes of vitamin A. J. Nutr. 113:2197– 2204. EFSA Panel on Additives and Products or Substances used in Animal Feed (FEEDAP). 2010. Scientific opinion on the safety and efficacy of vitamin E as a feed additive for all animal species. EFSA J. 8:1635. 10.2903/j. efsa.2010.1635. Eicher, S. D., J. L. Morrill, and J. Velazco. 1997. Bioavailability of α-tocopherol fed with retinol and relative bioavailability of d-αtocopherol or dl-α-tocopherol acetate. J. Dairy Sci. 80:393–399. Garrett, W. 1971. Energetic efficiency of beef and dairy steers. J. Anim. Sci. 31:452–456.
Gibb, D. J., F. H. Van Herk, P. S. Mir, S. Loerch, and T. A. McAllister. 2011. Removal of supplemental vitamin A from barley-based diets improves marbling in feedlot heifers. Can. J. Anim. Sci. 91:669–674.
Perry, T. W., W. M. Beeson, W. H. Smith, R. B. Harrington, and M. T. Mohler. 1968. Interrelationships among vitamins A, E, and K when added to the rations of fattening beef cattle. J. Anim. Sci. 27:190–194.
Gorocica-Buenfil, M. A., F. L. Fluharty, C. K. Reynolds, and S. C. Loerch. 2007a. Effect of dietary vitamin A concentration and roasted soybean inclusion on marbling, adipose cellularity, and fatty acid composition of beef. J. Anim. Sci. 85:2230–2242.
Pickworth, C. L., S. C. Loerch, R. E. Kopec, S. J. Schwartz, and F. L. Fluharty. 2012. Concentration of pro-vitamin A carotenoids in common beef cattle feedstuffs. J. Anim. Sci. 90:1553–1561.
Gorocica-Buenfil, M. A., F. L. Fluharty, C. K. Reynolds, and S. C. Loerch. 2007b. Effect of dietary vitamin A restriction on marbling and conjugated linoleic acid content in Holstein steers. J. Anim. Sci. 85:2243–2255. Hicks, C. R. 1973. Fundamental Concepts in the Design of Experiments. Holt, Rinehart and Winston Inc., New York, NY. Hidiroglou, N., L. F. Laflamme, and L. R. McDowell. 1988. Blood plasma and tissue concentrations of vitamin E in beef cattle as influenced by supplementation of various compounds. J. Anim. Sci. 66:3227–3234. Hidiroglou, N., L. R. McDowell, A. M. Papas, M. Antapli, and N. S. Wilkinson. 1992. Bioavailability of vitamin E compounds in lambs. J. Anim. Sci. 70:2556–2561. Kohlmeier, R. H., and W. Burroughs. 1970. Estimation of critical plasma and liver vitamin A levels in feedlot cattle with observation upon influences of body stores and daily dietary requirements. J. Anim. Sci. 30:1012–1018. Montgomery, S. P., J. S. Drouillard, J. J. Sindt, M. A. Greenquist, B. E. Depenbusch, E. J. Good, E. R. Loe, M. J. Sulpizio, T. J. Kessen, and R. T. Ethington. 2005. Effects of dried full-fat corn germ and vitamin E on growth performance and carcass characteristics of finishing cattle. J. Anim. Sci. 83:2440–2447. NRC. 1984. Nutrient Requirements of Beef Cattle. 6th rev. ed. Natl. Acad. Press, Washington, DC. NRC. 2000. Nutrient Requirements of Beef Cattle. Updated 7th rev. ed. Natl. Acad. Press, Washington, DC. Pehrson, B., J. Hakkarainen, M. Tornquist, K. Edfors, and C. Fossum. 1991. Effect of vitamin E supplementation on weight gain, immune competence, and disease incidence in barley-fed beef cattle. J. Dairy Sci. 74:1054– 1059.
Reddy, P. G., J. L. Morrill, and R. A. Frey. 1987. Vitamin E requirement of dairy calves. J. Dairy Sci. 70:123–129. Rivera, J. D., G. C. Duff, M. L. Galyean, D. A. Walker, and G. A. Nunnery. 2002. Effects of supplemental vitamin E on performance, health, and humoral immune response of beef cattle. J. Anim. Sci. 80:933–941. Rode, L. M., T. A. McAllister, and K. J. Cheng. 1990. Microbial degradation of vitamin A in rumen fluid from steers fed concentrate, hay, or straw diets. Can. J. Anim. Sci. 70:227–233. Stahr, H. M. 1991. Analytical Methods in Toxicology. John Wiley & Sons, Inc. New York, NY. Vagni, S., F. Saccone, L. Pinotti, and A. Baldi. 2011. Vitamin E bioavailability: Past and present insights. Food Nutr. Sci. 2:1088–1096. Weiss, W. P., J. S. Hogan, and D. J. Wyatt. 2009. Relative bioavailability of all-rac and RRR vitamin E based on neutrophil function and total alpha-tocopherol and isomer concentrations in periparturient dairy cows and their calves. J. Dairy Sci. 92:720–731. Zinn, R. A. 1987. Influence of lasalocid and monensin plus tylosin on comparative feeding value of steam-flaked versus dry-rolled corn diets for feedlot cattle. J. Anim. Sci. 65:256–266. Zinn, R. A., E. Alvarez, and R. L. Stuart. 1996. Interaction of supplemental vitamin A and E on health and performance of crossbred and Holstein calves during the receiving. Prof. Anim. Sci. 12:17–23. Zinn, R. A., and Y. Shen. 1998. An evaluation of ruminally degradable intake protein and metabolizable amino acid requirements of feedlot calves. J. Anim. Sci. 76:1280–1289.
Evaluation of supplemental vitamin A and E on 56-day
growth performance, dietary net energy, and plasma retinol and tocopherol concentrations in Holstein steer calves J. Salinas-Chavira,* A. A. Arrizon,† A. Barreras,† C. Z. Chen,‡ A. Plascencia,† and R. A. Zinn§ *Facultad de Medicina Veterinaria y Zootecnia, Universidad Autonoma de Tamaulipas, Cd. Victoria, Tam. 87000, México; †Instituto de Investigaciones en Ciencias Veterinarias, Universidad Autonoma de Baja California, Mexicali, Baja California 21100, México; ‡College of Animal Science, Jilin University, Changchun, China 130062; and §Department of Animal Science, University of California, Davis 95616
ABSTRACT Thirty Holsteins steers (initial weight 230 ± 1 kg) fed as calves were used in a 56-d experiment to evaluate the influence of vitamin A and E supplementation on growth performance, dietary energetics, and plasma retinol and tocopherol concentrations. Steers were fed a steamflaked, corn-based diet. Three treatments (TMT) were evaluated: TMT1 had no supplemental vitamins, TMT2 supplemented 30,000 IU/d of vitamin A as retinyl propionate plus 250 IU/d of vitamin E as all-rac α-tocopheryl acetate, and TMT3 supplemented 30,000 IU/d of vitamin A as retinyl palmitate plus 250 IU/d of vitamin E as d-α-tocopherol (RRR α-tocopherol). Treatment supple-
1 Corresponding author: razinn@ucdavis. edu
ments were freshly prepared and applied to feed once daily in the morning. Form of supplemental vitamins A and E did not affect cattle growth performance or dietary energetics. Vitamin supplementation increased final BW (3.7%, P = 0.02), ADG (10.7%, P = 0.02), and DMI (7.1%, P = 0.05). There were no treatment effects (P > 0.30) on gain efficiency or dietary net energy. There were no treatment effects on plasma retinol on d 28. However, by d 56, vitamin A supplementation increased (P = 0.01) plasma retinol by 56%. Form of supplemental vitamin A did not influence (P > 0.30) plasma retinol. Vitamin E supplementation increased (P < 0.01) plasma tocopherol. On d 28, plasma tocopherol tended to be greater (P = 0.12) with supplemental tocopheryl acetate than for supplemental tocopherol. By d 56 plasma tocopherol was 31% greater (P = 0.03) for all-rac α-tocopheryl acetate than for
RRR α-tocopherol supplemented steers. Vitamin supplementation may enhance DMI and corresponding ADG of Holstein steer calves fed a steam-flaked corn-based diet. Ester forms of vitamin A (retinyl propionate vs. retinyl palmitate) are comparable. Key words: feedlot, Holstein, vitamin A, vitamin E
INTRODUCTION Basal ingredients (grains and roughages) commonly used in growingfinishing diet formulations for feedlot cattle are characteristically deficient in supplying vitamin A and, possibly, vitamin E (NRC, 2000); therefore, dietary supplementation is a common practice. In addition to changes in growth performance, bioavailability of
511
Vitamins A and E on performance and plasma levels
supplemental vitamin A and E forms are often evaluated based on resulting changes in plasma concentrations, tissue concentrations, or both (Vagni et al., 2011). Response to vitamin A and E supplementation of feedlot cattle has been variable (Perry et al., 1968; Arnold et al., 1992; Bryant et al., 2010; Burken et al., 2012), due in part to cattle background and composition of the basal diet. There is limited information regarding the vitamin A and E status of Holstein steers fed as calves a conventional steam-flaked, corn-based, growing-finishing diet. The objective of the current experiment was evaluate vitamin A and E supplementation of Holstein steers fed as calves, comparing the combinations of retinyl propionate and all-rac α-tocopheryl acetate versus retinyl palmitate and RRR α-tocopherol.
MATERIALS AND METHODS All animal care, handling, and sample techniques followed protocols approved by the University of California, Davis, Animal Use and Care Committee.
Animals and Design Thirty Holstein steer calves (initial BW = 230 ± 1 kg) were used in a 56-d experiment to evaluate influence supplemental vitamin A and E on growth-performance, dietary NE, and plasma vitamin concentrations. Steers were blocked by initial BW and assigned within BW groupings to 15 pens, 2 steers/pen. Pens were 5.5 × 9.1 m with 27 m2 of shade and were equipped with automatic waterers and fence-line feed bunks (4.3-m long). Upon initiation, the steers were implanted with Revalor-S (Intervet Inc., Millsboro, DE). The current experiement used 3 dietary treatments (TMT) consisting of a steam-flaked, corn-based, growing-finishing diet (Table 1) and supplements: (TMT1) a control with no supplemental vitamins; (TMT2) 30,000 IU/d of vitamin A as retinyl propionate (Microvit A DLC 500 HF; Adisseo USA Inc.,
Table 1. Composition of experimental diets
Item
Basal diet, %
Ingredient composition (DM basis) Steam-flaked corn 62.96 Alfalfa hay 3.87 Sudangrass hay 7.74 2.67 Yellow grease Molasses cane 5.93 Distillers dried 14.36 0.58 Urea Limestone 1.45 Magnesium oxide 0.09 Monensin, mg/kg 30.00 Trace mineral salt1 0.35 Nutrient composition (DM basis)2 NE, Mcal/kg Maintenance 2.21 Gain 1.54 CP, % 13.91 Calcium, % 0.72 Phosphorus, % 0.35 Vitamin A, IU/kg 418 Vitamin E, IU/kg 25 Trace mineral salt contained: CoSO4, 0.068%; CuSO4, 1.04%; FeSO4, 3.57%; ZnO, 1.24%; MnSO4, 1.07%, KI 0.052%; and NaCl, 92.96%. 2 Based on tabular values for individual feed ingredients (NRC, 2000), with the exception of vitamin A which was estimated based on vitamin A concentration of feed ingredients reported by Pickworth et al. (2012). 1
Alpharetta, GA) plus 250 IU/d of vitamin E as all-rac α-tocopheryl acetate (Microvit E DLC 60 HF; Adisseo USA Inc.); and (TMT3) 30,000 IU/d of vitamin A as retinyl palmitate (Emcelle Vitamin A-Liquid; Stuart Products, Bedford, TX) plus 250 IU/d of vitamin E as RRR α-tocopherol (Emcelle Tocopherol; Stuart Products). Steers were provided ad libitum access to the experimental diets. Fresh feed was provided twice daily (0600 and 1400 h). Daily application of vitamin treatments was accomplished by dilution in 30 mL of water before top dressing onto feed provided at the 0600 h feeding.
At 28-d intervals, blood samples were collected via jugular puncture into heparinized tubes. The blood was immediately centrifuged at 1,070 × g for 10 min at 23°C. Plasma α-tocopherol and retinol analyses were conducted by the Veterinary Diagnostic Laboratory (College of Veterinary Medicine, Iowa State University, Ames) according to procedures described by Stahr (1991).
Estimation of dietary NE Daily energy gain (EG; Mcal/d) was calculated by the equation EG = ADG1.097 × 0.0557W0.75, where W is the mean shrunk BW (kg; NRC, 1984). Maintenance energy (EM) was calculated by the equation EM = 0.084W0.75 (Garrett, 1971). Dietary NEg was derived from NEm by the equation NEg = 0.877NEm – 0.41 (Zinn, 1987). Dry matter intake is related to energy requirements and dietary NEm according to the equation DMI = EM/NEm + EG/(0.877NEm − 0.41), and can be resolved for estimation of dietary NE by means of the quadratic formula
where x = NEm; a = −0.41EM; b = 0.877 EM + 0.41DMI + EG; and c = −0.877DMI (Zinn and Shen, 1998).
Statistical Design and Analysis For calculating steer performance, initial and final BW were reduced 4% to account for digestive tract fill. The experimental data were analyzed as a randomized complete block design according to the statistical model Yij = μ + Bi + Tj + εij (Hicks, 1973), where μ is the common experimental effect; Bi represents blocks (df = 4); Tj represents dietary treatment effect (df = 2), and εij represents the residual error (df = 8). Treatments effects were tested by means of orthogonal contrasts as (1) control versus supplemental vitamins and (2) TMT2 versus TMT3. Analysis was performed using
512
Salinas-Chavira et al.
Statistix-10 (Analytical Software, Tallahassee, FL).
vitamin A or E, or the combination of both. Perry et al. (1968) evaluated vitamin A and E supplementation of ground corn-based finishing diets fed to feedlot steer calves. Vitamins were fed either singly or in combination. Fed individually, vitamin A supplementation increased DMI and ADG, whereas vitamin E supplementation did not affect growth performance. Likewise, the combination of vitamin A and vitamin E supplementation did not affect growth performance beyond that observed for vitamin A supplementation. However, the effect of vitamin E supplementation of feedlot cattle on growth performance has not been consistent. Whereas numerous subsequent studies (Arnold et al., 1992; Zinn et al., 1996; Rivera et al., 2002; Carter et al., 2005; Montgomery et al., 2005) have likewise failed to show a growth performance response to supplemental vitamin E, in other instances vitamin E supplementation enhanced daily BW gain (Reddy et al., 1987; Pehrson et al., 1991; Burken et al., 2012). Daily BW gain response to vitamin E supplementation is apparent when plasma tocopherol
RESULTS AND DISCUSSION Effects of Treatments on Growth Performance and Dietary Energetics Treatment effects on cattle growthperformance and dietary NE are shown in Table 2. Form of supplemental vitamins A (retinyl propionate vs. retinyl palmitate) and E (RRR α-tocopherol vs all-rac α-tocopheryl acetate) did not affect cattle growth performance or dietary energetics. However, vitamin supplementation increased final BW (3.7%, P = 0.02), ADG (10.7%, P = 0.02), and DMI (7.1%, P = 0.05). The improvement in ADG is attributable to increased DMI, as there were no treatment effects (P > 0.30) on BW gain efficiency or dietary net energy. It is not possible in the present experiment to determine if beneficial treatment effects on growth performance were due to supplemental
levels are less than 1 μg/mL (Reddy et al., 1987; Pehrson et al., 1991). In the present experiment the minimum observed plasma tocopherol concentration of nonsupplemented steers was 1.4 μg/mL (Table 3). Kohlmeier and Burroughs (1970) observed that the critical level of plasma retinol below which supplementation enhanced ADG was 0.25 μg/mL. This observation is consistent with recent studies (Gorocica-Buenfil et al., 2007a,b; Gibb et al., 2011) where vitamin A supplementation did not enhance feedlot cattle growthperformance. Although, in an experiment reported by Bryant et al. (2010) where initial plasma retinol average 0.15 μg/mL, vitamin A supplementation also failed to enhance daily BW gain. However, in their study, plasma retinol of nonsupplemented steers increased with days on feed. In the present experiment, initial plasma retinol of nonsupplemented steers was 0.23 μg/mL and decreased with days on feed (Table 3), suggestive of marginal vitamin A stores in our Holstein steers fed as calves.
Table 2. Growth performance of Holstein calves that received 2 forms of vitamin A and E Treatment
Item Pen replications Live BW,4 kg Initial Final ADG, kg/d DMI, kg/d G:F Dietary NEm, Mcal/kg Dietary NEg, Mcal/kg Observed/expected NEm Observed/expected NEg
P-value
Control1
AE12
AE23
5
5
5
229 322 1.67 7.26 0.22 2.13 1.46 0.96 0.95
231 333 1.82 7.63 0.23 2.18 1.51 0.98 0.98
229 336 1.91 8.01 0.23 2.17 1.49 0.97 0.97
Control vs. supplemental vitamins
AE1 vs. AE2
SEM
0.53 0.02 0.02 0.05 0.32 0.50 0.50 0.50 0.50
0.310 0.541 0.330 0.216 0.990 0.825 0.825 0.825 0.825
1 3 0.05 0.18 0.01 0.05 0.04 0.02 0.03
Control = no supplemental vitamins. AE1 = vitamin A and E combination consisting of a daily dosage of 30,000 IU of vitamin A as retinyl propionate plus 250 IU of vitamin E as all-rac α-tocopheryl acetate. 3 AE2 = vitamin A and E combination consisting of a daily dosage of 30,000 IU of vitamin A as retinyl palmitate plus 250 IU of vitamin E as RRR α-tocopherol. 4 Body weights were reduced 4% to account for digestive tract fill. 1 2
513
Vitamins A and E on performance and plasma levels
Table 3. Treatments effects on plasma retinol and tocopherol concentrations of calves that received 2 forms of vitamin A and E Treatment
Item
Control1
Pen replications Plasma retinol, µg/mL 0d 28 d 56 d Plasma tocopherol, µg/mL 0d 28 d 56 d
P-value
AE12
AE23
5
5
5
0.23 0.21 0.16
0.21 0.23 0.26
2 2.04 1.36
1.81 3.84 3.30
0.23 0.25 0.25 2.17 3.31 2.52
Control vs. supplemental vitamins
0.59 0.15 0.01 0.97 0.00 0.00
TM2 vs. TMT3
0.36 0.38 0.78 0.24 0.12 0.03
SEM
0.01 0.01 0.02 0.18 0.20 0.18
Control = no supplemental vitamins. AE1 = vitamin A and E combination consisting of a daily dosage of 30,000 IU of vitamin A as retinyl propionate plus 250 IU of vitamin E as all-rac α-tocopheryl acetate . 3 AE2 = vitamin A and E combination consisting of a daily dosage of 30,000 IU of vitamin A as retinyl palmitate plus 250 IU of vitamin E as RRR α-tocopherol. 1 2
Effects of Treatments on Plasma Level of Vitamin A and E Treatment effects on plasma retinol and tocopherol concentrations are shown in Table 3. There were no effects of treatments on plasma retinol on d 28. However, by d 56, vitamin A supplementation increased (P = 0.01) plasma retinol by 56%. Form of supplemental vitamin A (retinyl propionate vs. retinyl palmitate) did not influence (P > 0.30) plasma retinol concentrations. Comparison of the 2 forms of vitamin A on plasma retinol concentrations has not been previously reported. Based on in vitro incubation research using ruminal fluid from steers fed a high-concentrate diet, Rode et al. (1990) observed that supplemental vitamin A (retinyl propionate) is extensively degraded by ruminal fermentation. After 12 h of incubation, 72% of retinol had been degraded, with degradation of retinol proceeding at a faster rate than hydrolysis of the retinyl ester. Based on an assumed ruminal dilution rate of 5%/h, they estimated that 33% of supplemental vitamin A reaches the small intestine of cattle
fed a high-concentrate diet. Assuming that 90% of vitamin A entering the small intestine is absorbed (Donoghue et al., 1983), then it is expected that approximately 30% of supplemental vitamin A was metabolizable. As expected (Pehrson et al., 1991; Arnold et al., 1992, 1993; Rivera et al., 2002), vitamin E supplementation increased (P < 0.01) plasma tocopherol concentration. On d 28, plasma tocopherol tended to be 16% greater (P = 0.12) with supplemental tocopheryl acetate than for supplemental tocopherol. By d 56, plasma tocopherol was 31% greater (P = 0.03) for all-rac α-tocopheryl acetate-supplemented steers than for RRR α-tocopherolsupplemented steers. This later result was unexpected. Based on standards of bioavailability (EFSA, 2010), the potency of RRR α-tocopherol is 49% greater than that of all-rac α-tocopheryl acetate. Accordingly, Hidiroglou et al. (1988) and Eicher et al. (1997) observed greater plasma tocopherol concentrations in cattle orally dosed with RRR α-tocopherol than with the all-rac α-tocopheryl acetate. In research using sheep (Hidiroglou et al., 1992) or dairy cattle (Weiss et al., 2009), dietary supplementation with
RRR α-tocopheryl acetate resulted in greater plasma tocopherol concentrations than supplementation with all-rac α-tocopheryl acetate. Comparative studies evaluating dietary supplementation of the free alcohol form of vitamin E have not been previously reported. However, the free alcohol form (RRR α-tocopherol) is more susceptible to oxidation than the ester forms (Vagni et al., 2011). Thus, although vitamin treatments were freshly applied to the feed daily, lower plasma tocopherol concentrations may reflect oxidative loss of the RRR α-tocopherol before consumption. More research is needed to evaluate differences in oxidative stability of vitamin E forms in growing-finishing diets for cattle.
IMPLICATIONS Vitamin supplementation may enhance DMI and corresponding ADG of Holstein steer calves fed a steamflaked, corn-based, growing-finishing diet. Ester forms of vitamin A (retinyl propionate vs. retinyl palmitate) are comparable. Based on plasma tocopherol levels, the basal nonsupplemented diet was not deficient in vitamin E.
514
Salinas-Chavira et al.
LITERATURE CITED Arnold, R. N., S. C. Arp, K. K. Scheller, S. N. Williams, and D. M. Schaefer. 1993. Tissue equilibration and subcellular distribution of vitamin E relative to myoglobin and lipid oxidation in displayed beef. J. Anim. Sci. 71:105–118. Arnold, R. N., K. K. Scheller, S. C. Arp, S. N. Williams, D. R. Buege, and D. M. Schaefer. 1992. Effect of long- or short-term feeding of α-tocopheryl acetate to Holstein and crossbred beef steers on performance, carcass characteristics, and beef color stability. J. Anim. Sci. 70:3055–3065. Bryant, T. C., J. J. Wagner, J. D. Tatum, M. L. Galyean, R. V. Anthony, and T. E. Engle. 2010. Effect of dietary supplemental vitamin A concentration on performance, carcass merit, serum metabolites, and lipogenic enzyme activity in yearling beef steers. J. Anim. Sci. 88:1463–1478. Burken, D. B., R. B. Hicks, D. L. VanOverbeke, G. G. Hilton, J. L. Wahrmund, B. P. Holland, C. R. Krehbiel, P. K. Camfield, and C. J. Richards. 2012. Vitamin E supplementation in beef finishing diets containing 35% wet distillers grains with solubles: Feedlot performance and carcass characteristics. J. Anim. Sci. 90:1349–1355. Carter, J. N., D. R. Gill, C. R. Krehbiel, A. W. Confer, R. A. Smith, D. L. Lalman, P. L. Claypool, and L. R. McDowell. 2005. Vitamin E supplementation of newly arrived feedlot calves. J. Anim. Sci. 83:1924–1932. Donoghue, S., W. J. Donawick, and D. S. Kronfeld. 1983. Transfer of vitamin A from intestine to plasma in lambs fed low and high intakes of vitamin A. J. Nutr. 113:2197– 2204. EFSA Panel on Additives and Products or Substances used in Animal Feed (FEEDAP). 2010. Scientific opinion on the safety and efficacy of vitamin E as a feed additive for all animal species. EFSA J. 8:1635. 10.2903/j. efsa.2010.1635. Eicher, S. D., J. L. Morrill, and J. Velazco. 1997. Bioavailability of α-tocopherol fed with retinol and relative bioavailability of d-αtocopherol or dl-α-tocopherol acetate. J. Dairy Sci. 80:393–399. Garrett, W. 1971. Energetic efficiency of beef and dairy steers. J. Anim. Sci. 31:452–456.
Gibb, D. J., F. H. Van Herk, P. S. Mir, S. Loerch, and T. A. McAllister. 2011. Removal of supplemental vitamin A from barley-based diets improves marbling in feedlot heifers. Can. J. Anim. Sci. 91:669–674.
Perry, T. W., W. M. Beeson, W. H. Smith, R. B. Harrington, and M. T. Mohler. 1968. Interrelationships among vitamins A, E, and K when added to the rations of fattening beef cattle. J. Anim. Sci. 27:190–194.
Gorocica-Buenfil, M. A., F. L. Fluharty, C. K. Reynolds, and S. C. Loerch. 2007a. Effect of dietary vitamin A concentration and roasted soybean inclusion on marbling, adipose cellularity, and fatty acid composition of beef. J. Anim. Sci. 85:2230–2242.
Pickworth, C. L., S. C. Loerch, R. E. Kopec, S. J. Schwartz, and F. L. Fluharty. 2012. Concentration of pro-vitamin A carotenoids in common beef cattle feedstuffs. J. Anim. Sci. 90:1553–1561.
Gorocica-Buenfil, M. A., F. L. Fluharty, C. K. Reynolds, and S. C. Loerch. 2007b. Effect of dietary vitamin A restriction on marbling and conjugated linoleic acid content in Holstein steers. J. Anim. Sci. 85:2243–2255. Hicks, C. R. 1973. Fundamental Concepts in the Design of Experiments. Holt, Rinehart and Winston Inc., New York, NY. Hidiroglou, N., L. F. Laflamme, and L. R. McDowell. 1988. Blood plasma and tissue concentrations of vitamin E in beef cattle as influenced by supplementation of various compounds. J. Anim. Sci. 66:3227–3234. Hidiroglou, N., L. R. McDowell, A. M. Papas, M. Antapli, and N. S. Wilkinson. 1992. Bioavailability of vitamin E compounds in lambs. J. Anim. Sci. 70:2556–2561. Kohlmeier, R. H., and W. Burroughs. 1970. Estimation of critical plasma and liver vitamin A levels in feedlot cattle with observation upon influences of body stores and daily dietary requirements. J. Anim. Sci. 30:1012–1018. Montgomery, S. P., J. S. Drouillard, J. J. Sindt, M. A. Greenquist, B. E. Depenbusch, E. J. Good, E. R. Loe, M. J. Sulpizio, T. J. Kessen, and R. T. Ethington. 2005. Effects of dried full-fat corn germ and vitamin E on growth performance and carcass characteristics of finishing cattle. J. Anim. Sci. 83:2440–2447. NRC. 1984. Nutrient Requirements of Beef Cattle. 6th rev. ed. Natl. Acad. Press, Washington, DC. NRC. 2000. Nutrient Requirements of Beef Cattle. Updated 7th rev. ed. Natl. Acad. Press, Washington, DC. Pehrson, B., J. Hakkarainen, M. Tornquist, K. Edfors, and C. Fossum. 1991. Effect of vitamin E supplementation on weight gain, immune competence, and disease incidence in barley-fed beef cattle. J. Dairy Sci. 74:1054– 1059.
Reddy, P. G., J. L. Morrill, and R. A. Frey. 1987. Vitamin E requirement of dairy calves. J. Dairy Sci. 70:123–129. Rivera, J. D., G. C. Duff, M. L. Galyean, D. A. Walker, and G. A. Nunnery. 2002. Effects of supplemental vitamin E on performance, health, and humoral immune response of beef cattle. J. Anim. Sci. 80:933–941. Rode, L. M., T. A. McAllister, and K. J. Cheng. 1990. Microbial degradation of vitamin A in rumen fluid from steers fed concentrate, hay, or straw diets. Can. J. Anim. Sci. 70:227–233. Stahr, H. M. 1991. Analytical Methods in Toxicology. John Wiley & Sons, Inc. New York, NY. Vagni, S., F. Saccone, L. Pinotti, and A. Baldi. 2011. Vitamin E bioavailability: Past and present insights. Food Nutr. Sci. 2:1088–1096. Weiss, W. P., J. S. Hogan, and D. J. Wyatt. 2009. Relative bioavailability of all-rac and RRR vitamin E based on neutrophil function and total alpha-tocopherol and isomer concentrations in periparturient dairy cows and their calves. J. Dairy Sci. 92:720–731. Zinn, R. A. 1987. Influence of lasalocid and monensin plus tylosin on comparative feeding value of steam-flaked versus dry-rolled corn diets for feedlot cattle. J. Anim. Sci. 65:256–266. Zinn, R. A., E. Alvarez, and R. L. Stuart. 1996. Interaction of supplemental vitamin A and E on health and performance of crossbred and Holstein calves during the receiving. Prof. Anim. Sci. 12:17–23. Zinn, R. A., and Y. Shen. 1998. An evaluation of ruminally degradable intake protein and metabolizable amino acid requirements of feedlot calves. J. Anim. Sci. 76:1280–1289.