Selenium/Vitamin E: Role in Disease Prevention and Weight Gain of Neonatal Calves 1'2

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1 Selenium/Vitamin E: Role in Disease Prevention and Weight Gain of Neonatal Calves 1'2 W. P. WEISS, 3 V. F. COLENBRANDER, 3 M. D. CUNNINGHAM, 3 and C. J. CALLAHAN 4 Purdue University West Lafayette, IN ABSTRACT A 3 2 factorial arrangement of treatments comparing 0, 1, or 2 injections of a selenium/vitamin E preparation and feeding of starter commencing at 14 or 28 days of age was used to examine effects of supplemental selenium/vitamin E on health and growth of neonatal Dosage per injection was.078 mg selenium and 5.4 international units of vitamin E per kg body weight. Neither a single injection at birth nor two injections (birth and day 14) reduced incidence of respiratory diseases or improved weight gains in unstressed Calves first offered starter at 14 days as compared with 28 days gained more weight in the 56 days. A single injection of selenium/ vitamin E at birth increased selenium of blood serum at day 14 by 87%, and a second injection increased selenium of blood serum at day 28 an additional 10%. Selenium of blood serum of calves not given an injection decreased linearly from birth to day 56. Calves first fed starter at day 14 had lower concentrations of selenium of blood serum from day 28 to 42 than calves first offered starter at day 28. Glutathione peroxidase activity of erythrocytes was higher in calves receiving one or two injections of selenium/ vitamin E than in control calves; however, a lag of 4 to 5 wk occurred between injections and elevation of glutathione peroxidase activity. Received July 19, I Journal Paper No. 9114, Purdue University Agricultural Experiment Station. ~Research supported in part by Indiana Farm Bureau Cooperative, Inc. 3 Department of Animal Sciences. 4 Large Animal Clinic, School of Veterinary Medicine. INTRODUCTION Selenium and vitamin E are essential micronutrients required for good health of animals throughout their life cycle and for attainment of high production (10). The occurrence of Se/vit E-responsiveness diseases has increased because of substitution of stored feedstuffs for pasture and because fewer concentrates grown in areas having adequate amounts of selenium in the soil are fed now (10). The National Research Council has established the selenium requirement for all classes of dairy cattle at.1 ppm of total dry matter intake (16). Crops grown adjacent to the Great Lakes are low in selenium (2). More than 80% of forages and grain crops grown locally and fed to dairy cattle in Indiana contain less than.05 ppm of selenium (13, 21). Milk produced and consumed by calves in this area is exceedingly low in selenium content (4, 21). Both selenium and vitamin E are required by animals; however, increased intake of one of these nutrients may decrease the amount of the other nutrient needed to prevent deficiency diseases. Currently there are two explanations for this interrelationship. Vitamin E, which functions as an antioxidant, may help maintain selenium in the reduced or active state, thereby decreasing the amount of selenium required (5). Vitamin E also may protect selenium activity from interference by lipids (10). One biochemical function of selenium is as a component of the enzyme, glutathione peroxidase (22). Selenium-dependent glutathione peroxidase (Se-GSHpx) protects cell membranes from oxidative damage produced by peroxides derived from unsaturated fatty acids within ceils (23). Activity of Se-GSHpx in certain tissues may be useful for establishing the selenium status of animals (17). Mortality from respiratory and gastrointestinal diseases often is high for neonatal Feeding supplemental selenium improved weight gains and increased resistance to respira J Dairy Sci 66:

2 1102 WEISS ET AL. tory diseases and scours in milk-fed calves (15). However, supplemental dietary selenium had no effect on weight gains in calves in (26). The objective of this experiment was to evaluate effects of injected Se and vitamin E on health and weight gains of calves from dams with low selenium in blood serum that were fed selenium deficient diets. MATERIALS AND METHODS Forty-two Holstein heifer calves (six/treatment) were allotted to one of seven blocks according to birth order. One calf from each block was assigned randomly to one of six treatments in a 3 2 factorial arrangement of treatments. Treatments were 0, 1, or 2 injections of Se/vit E and feeding of calf starter commencing at either day 14 or day 28. The Se/vit E s was administered subcutaneously at birth (one injection) or at birth and day 14 (two injections). Calves less than 41 kg in body weight at birth received 3.0 ml, those weighing 41 to 45.4 kg received 3.4 ml, and calves weighing more than 45.4 kg received 3.75 ml per injection. Dams had an average of 15 ppb Se in blood serum at parturition. Each calf received 900 g colostrum from its dam within 2 h of birth and then was fed 3.6 kg colostrum per day for 3 days. From day 4 to day 35, calves received milk replacer powder 6 mixed with warm water, 1:9 wt/wt fed in two equal feedings to total 9% of body weight daily. Between day 36 and weaning at day 42, milk replacer was mixed at a ratio of 1:18. Milk replacer was unmedicated and contained no supplemental Se (basal.068 ppm) or vitamin E. Guaranteed analysis of the replacer was not less than 21% crude protein (half derived from soy protein concentrate), 15% crude fat, 33 USP units vitamin A per g, 6.6 USP units vitamin D3 per g and not more than.6% crude fiber. Formulation of calf starter is in Table 1. The estimated proximate analysis of this calf starter was 88.7% dry matter (DM), 16.5% crude protein, 6.1% crude s BO-SE was supplied by Burns-Biotec Laboratories, Oakland, CA Each milliliter contained 1 mg selenium equivalent from sodium selenite plus 68 IU vitamin E as d-alpha tocopheryl acetate. 6 Milk replacer was specifically formulated and supplied by Milk Specialities Co., Dundee, IL TABLE 1. Formulation of calf starter, a Ingredient (kg/loo kg) Cracked yellow corn 36.6 Crimped oats 36.6 Soybean oil meal (44% CP) 17.9 Dried molasses 6.1 Dicalcium phosphate 1.1 Ground limestone 1.4 Salt, trace mineralized.2 Vitamin A and D premix b.y adry matter basis bpremix contained 12,587,000 USP vitamin A and 3,147,000 USP vitamin D3/kg. Starter contained.060 ppm Se (dry weight). fiber, 3.4% ether extract, 57.3% nitrogen-free extract,.8% calcium, and.6% phosphorus. Calves were fed starter according to appetite during the milk feeding stage. Neither water nor hay was available during the preweaning period. Calves were assigned by blocks to either conventional calf housing with forced-air ventilation and supplemental heat at 10 C or to cold-type housing consisting of small sheds. All calves were maintained in straw-bedded individual pens, approximately 1.2 x 1.8 m. Daily observations of calves were made for scours and respiratory ailments. Calves were not vaccinated for any specific disease during the experiment. At weaning, calves were moved to community pens of four to six Following weaning, calves received hay and water ad libitum and starter at 1.4 kg per calf daily. Body weights were obtained at birth and then once weekly throughout the 8 wk. Two blood samples were collected from each calf in vacutainers by jugular venipuncture at birth, days 14, 28, 35, 42, and 56. One 10-ml sample was allowed to coagulate and centrifuged to separate serum from cells. The resultant serum was frozen in sterile 5-ml polypropylene tubes until assayed fluorometrically for selenium by the procedure of Olson et al. (19) as modified by A. L. Moxon (personal communication). A 5-ml blood sample containing EDTA was prepared according to the method of Paglia and Valentine (20) for analysis of glutathione peroxidase activity. Blood was centri-

3 SELENIUM/VITAMIN E FOR CALVES 1103 fuged at 2000 rpm for 15 min to separate erythrocytes (RBC) from plasma. The RBC then were washed.three times by resuspending erythrocytes in 4 ml of saline phosphate soluti6n and centrifuging at 2000 rpm for 15 min. Finally,.4 ml of RBC was pipetted into plastic storage tubes containing 1.6 ml of distilled water. The hemolysate was frozen and stored until assayed for Se-GSHpx activity by the method of Paglia and Valentine (20) as modified by Hoekstra (personal communication). Activity of Se-GSHpx was measured as enzyme units (eu) per mg hemoglobin (Hb). An eu was equal to change in log [GSH]" 103-min -1. Analysis of variance and regression techniques were used to interpret data. When significant effects were found, means were compared by either Fisher's modified least significant difference test (unequal cell size) or Duncan's multiple range test (28). Health RESULTS AND DISCUSSION There was no difference in either incidence or duration of scours between calves given 0, 1, or 2 injections of Se/vit E (Table 2). Feeding supplemental Se to deficient calves may help prevent scours (15, 18). Selenium supplementation increased serum IgM concentrations in mice (27) and maintained integrity of intestinal mucosa in calves (15). In the calf, IgM helps prevent enteritis (14), which may be the connection between Se supplementation and reduced incidence of scours of some reports. However, Ferris and Thomas (6) reported that good management practices may decrease the importance of high serum IgM in disease prevention. During our study health problems were minimal. Calves first offered starter at day 28 (late starter, LS), instead of day 14 (early starter, ES) tended to have fewer cases of scours. Incidence of scours was not associated with any particular period of feeding. There is evidence that calves fed at maintenance for the first 2 to 3 wk of life have a lower incidence of scours than do calves fed for growth during that period (11). Ohio workers (15) found a lower incidence of respiratory ailments in Se-deficient calves fed supplemental Se than in calves fed no supplemental Se. In our study too few problems were encountered to make any conclusive statements on the value of Se/vit E in the prevention of respiratory problems. Weight Gains and Starter Intake Treatment with Se/vit E had no effect on rate of gain, total gain, and starter intake (Table 3). There was no interaction between Se/vit E treatment and starter treatment for any performance variable. In contrast, Moser et al. (15) found that whole milk supplemented with.15 ppm Se significantly increased weight gain of Age when starter first was offered had a significant effect on weight gain. Both groups of calves (ES and LS) lost weight during the first 2 wk of life. Calves offered dry feed at 14 days (ES) started to gain weight at that time, whereas calves offered starter at day 28 (LS) continued to lose weight until day 21. At the conclusion of the 56 day trial ES calves TABLE 2. Scours and respiratory problems in calves injected with Se/vit E and starter offered at different ages. a Day starter offered Se/vit E injections Item Cases of scours Scours, days/calf Cases of respiratory congestion Cases of pneumonia asevere scours and respiratory ailments were diagnosed and treated by veterinarians. Calves with respiratory congestion were coughing, had nasal discharge, etc., but were not diagnosed as having pneumonia.

4 1104 WEISS ET AL. weighed 4.5 kg more than LS calves (P<.01) with net gains of 19 and 14 kg (Table 3). Average daily gains (ADG) were similar for both starter treatments for the first 14 days of life and for the last 28 days of the experiment (days 28 to 56). Therefore, the difference in weight gains can be attributed mainly to the 14 days when ES calves received starter while LS calves did not. The ADG between 14 and 28 days was,32 and.10 kg for ES and LS calves (P<.01) which accounted for 62% of the difference of net gain. This suggested that calves fed limited milk replacer (9% of body weight) need dry feed early for optimal rumen development and growth by weaning age. Hodgson (8) reported that calves offered dry feed at either day 4 or weaning (day 21) did not differ in total weight gains (day 62) when fed milk replacer ad libitum. Unrestricted feeding of milk replacer, however, provides little stimulus to the calf to consume dry feed and probably was the principal reason offering dry feed did not influence weight gain in the Hodgson study. Consumption of starter increased linearly from the time it was first offered (day 14 or 28) until day 42 for both starter treatment groups. Calves offered starter at day 28 increased consumption of starter more rapidly (P<.05) than did calves offered dry feed at day 14. Slopes of regression lines of kg starter intake per 100 kg body weight over weeks were.28 and.43 (r and.88) for ES and LS This implied that the increased need for nutrients by the calf shortened the time needed for LS calves to become accustomed to dry feed. Serum Selenium At birth, blood serum Se of calves averaged 22 to 24 ppb (Table 4). Selenium in serum of calves not injected (Se-0) decreased linearly TABLE 3. Effect of Se/vit E treatment and age starter offered on performance of Day starter offered Main effects a Se/vit E injections b Item SE SE No. of calves Body weight (kg) Birth Day 42 (weaned) 52.1 c 48.1 d Day c 57.4 d Total gain (kg) 0-42 days 9.3 c 4.7 d days 19.0c 14.0 d Average daily gain (kg/day) 0-14 days days.32 c.10 d days days Starter intake (kg dry matter/day) days days.52 c 129 d alnteraetion of Se/vit E X starter not significant (P>.lS). bse/vit E main effect not significant (P>.20). C'dMeans in same row differ (P<.01).

5 SELENIUM/VITAMIN E FOR CALVES 1105 (r 2 =.55) from 24 ppb at birth to 18.8 ppb at day 56. Selenium in serum of calves given one injection of Se/vit E at birth (Se-1) increased 85% from birth to day 14, then decreased quadratically (y = X +.009X2; r 2 =.59) and approached original birth concentration by day 56. Serum Se of Se-1 calves was consistendy higher (P<.01) than that of Se-O calves after injection anff during the remainder of the trial. The increase in serum Se at day 14 of calves injected at birth and day 14 (Se-2) was similar to that of Se-1 The second injection of Se/vit E given Se-2 calves did not increase serum Se as much as the first injection (90 vs 10% increase). These results support Kincaid et al. (12) who reported an inverse relationship between retention of 7SSe in blood and pretreatment concentration of Se in blood. As shown in Table 4 calves first fed starter at day 14 (ES) had lower concentrations of Se in blood serum on days 28 (P<.01), 35 (P<.01), and 42 (P<.05) than did calves first offered starter on day 28 (LS). This encompassed the period when ES calves were gaining weight more rapidly than LS calves (Table 3) and, therefore, may have been utilizing systemic Se faster for tissue growth and metabolism. Glutathione Peroxidase At birth, glutathione peroxidase activity in erythrocytes (RBC) averaged 14.9 eu/mg Hb for all calves (Table 5) which is similar to the report by Scholz et al. (25) for newborn The Se-GSHpx activity increased slightly for Se-0 calves from birth until day 28, followed by a decrease to near birth activity by day 56. No significant linear or quadratic time trend was evident (P>.10). Calves receiving one injection of Se/vit E showed a quadratic response (P<.01) of Se-GSHpx activity in RBC over time (y = X X2; r ) with a peak of 30.8 eu/mg Hb on day 35. From day 35 through day 56, Se-GSHpx activity in Se-1 calves was significantly higher than that of Se-0 calves (P<.05). Hoffman et al. (9) also reported a significant quadratic time effect of Se-GSHpx activity of dairy heifers given one injection of 5 mg Se per 60 kg body weight. Glutathione peroxidase activity in those heifers also peaked 35 days postinjection but lower (2 3 eu/mg Hb) than in Se-1 calves in this study. Activity of Se-GSHpx of Se-2 calves also showed a significant (P<.01) quadratic time effect (y = X X2; r 2 =.69). The observed maximum occurred between day 35 and 56, but regression analysis estimated the peak at day 56. At day 28, Se-2 calves had activity higher than control calves (P<.01), and from day 35 through day 56 Se-GSHpx activity of Se-2 calves remained higher (P<.05) than TABLE 4. Effect of Se/vit E treatment and age starter offered on concentration of selenium in serum. Day starter offered Main effect a Se/vit E injections Item SE SE No. of calves Serum Se, ppb Birth Day f 41.1g 41.9g 1.49 Day b 35.1 c f 33.1g 46.2 h 1.19 Day b 30.8 c f 27.8g 40.4 h.84 Day d 29.1 e f 26.4g 37.9 h.99 Day f 23.8g 31.7 h 1.03 alnteraction of Se/vit E starter not significant (P>.20). b'cmeans in same row within starter treatment with unlike superscripts differ (P<.01). d'emeans in same row within starter treatment with unlike superscripts differ (p<.05). f'g'hmeans in same row within Se/vit E treatment with unlike superscripts differ (P<.01).

6 1106 WEISS ET AL. both Se-0 and Se-1 Even though both serum Se and Se-GSHpx activity were elevated by injecting Se, no growth response or decrease in health problems were observed in treated Age at which calves first were offered starter had no effect on RBC glutathione peroxidase activity, and there was no significant interaction between starter and selenium treatment. Relationship Between Selenium and Se-GSHpx Activity Erythrocyte Se-GSHpx activity is correlated with total blood Se in cattle (1, 24). Hence, it has been postulated that Se-GSHpx can be used as a measure of the long-term Se status of animals (7). In the current study of all calves, there was no correlation between serum Se and erythrocyte Se-GSHpx activity from birth through day 28, but for the remainder of the experiment correlations were significant (P<.01). Coefficients of determination (r 2) between serum Se and Se-GSHpx for days 35, 42, and 56 were.61,.65, and.54. Serum of both Se-1 and Se-2 calves contained more (P<.01) Se than control calves at day 14; however, Se-GSHpx activity was not elevated significantly above control calves until days 28 through 35. Serum Se is elevated almost immediately following injection (12) which indicates a lag period of 4 to 5 wk between increase of serum Se and increase of Se- GSHpx activity. This agrees with Scholz and Hutchinson (24), who reported a 5-wk lag period between Se supplementation and increased erythrocyte Se-GSHpx activity of neonatal Serum Se of Se-1 calves decreased 32.4% from day 14 to day 25 whereas it decreased only 14.4% from day 35 to day 56. This indicated that less Se from the serum was being utilized and excreted during the last 21 days of the experiment than during days 14 through 35. When the rate of disappearance of Se from the serum started decreasing, Se-GSHpx activity of RBC also started to decline (day 35). One possible explanation is that calves were growing; thus, their total blood volume was increasing. At the same time, however, Se in serum was becoming limiting, and less Se was available for incorporation into Se-GSHpx during erythropoiesis. Therefore, RBC synthesized after the point where serum Se became limiting, contained less Se-GSHpx. This tended to dilute the total erythrocyte Se-GSHpx pool. Our results agree with Anderson et al. (3), who TABLE 5. Effect of Se/vit E treatment and age starter offered on erythrocyte glutathione peroxidase activity of Day starter offered b Main effects a Se/vit E injections Item SE SE No. of calves Se-GSHpx activity (eu/mg Hb) Birth Day Day c 25.6c, d 31.8 d 1.70 Day c, x 30.sd,y 39.0d, z 1.39 Day c 28.5 d 38.8 e 1.21 Day c, x 24.1c,y 38.9d, z 1.39 alnteraction of Se/vit E X starter not significant (P>.20). bstarter treatment not significant (P>.25). c'd'emeans in same row within Se/vit E treatment with unlike superscripts differ (P<.O1). x'y'zmeans in same row within Se/vit E treatment with unlike superscripts differ (P<.05).

7 SELENIUM/VITAMIN E FOR CALVES 1107 reported that Se-GSHpx activity of RBC of cattle decreased whereas total blood Se remained relatively constant. They also suggested that at a certain point Se becomes limiting and unavailable for incorporation into the enzyme. With Se-1 calves the threshold or limiting concentration was 28 ppb Se in serum. Moreover, if one extrapolates the regression curves (which is not recommended) for serum Se and Se-GSHpx activity of Se-2 calves, Se- GSHpx activity started to decrease noticeably when serum Se approached 28 ppb. Longer experiments are necessary to identify more conclusively the threshold for serum Se in REFERENCES I Allen, W. M., W. H. Parr, P. H. Anderson, S. Berrett, R. Bradley, and D.S.P. Patterson Selenium and the activity of glutathione peroxidase in bovine erythrocytes. Vet. Rec. 96: Ammerman, C. B., and S. M. Miller Selenium in ruminant nutrition: A review. J. Dairy Sci. 58: Anderson, P. H., S. Barrett, and D.S.P. Patterson Glutathione peroxidase activity in erythrocytes and muscle and cattle and sheep and its relationship to selenium. J. Comp. Pathol. 88: Conrad, H. R., and A. L. Moxon Transfer of dietary selenium to milk. J. Dairy Sci. 62: Diplock, A. T Page 190 in Proc. 1st Int. Syrup. Trace Minerals. Metab. Anita. C. F. Mills, ed. Livingston, Edinburgh. 6 Ferris, T. A., and J. W. Thomas Relationship of immunoglobulin to dairy calf mortality and influence of herd environment. J. Dairy Sci. 57: Hafeman, D. G., R. A. Sunde, and W. G. Hoekstra Effect of dietary selenium on erythrocyte and liver glutathione peroxidase in the rat. J. Nutr. 104: Hodgson, J The development of solid food intake in I. The effect of previous experience of solid food, and the physical form of the diet on the development of food intake after weaning. Anita. Prod. 13:15. 9 Hoffman, C., B. Rivinus, and L. Swanson Effect of intramuscular administration of selenium and vitamin E in dairy heifers on erythrocyte glutathione peroxidase activity and blood selenium levels. J. Anim. Sci. 47: Jenkins, K. J., and M. Hidiroglou.,1972. A review of selenium/vitamin E responsive problems in livestock: A case for selenium as a feed additive in Canada. Can. J. Anita. Sci. 52: Jenny, B. F., S. E. Mills, W. E. Johnston, and G. D. O'Dell Effect of fluid intake and dry matter concentrations on scours and water intake in calves fed once daily. J. Dairy Sci. 61: Kincaid, R. L., W. J. Miller, M. W. Neathery, R. P. Gentry, and D. L. Hampton Effect of added dietary selenium on metabolism and tissue distribution of radioactive and stable selenium in J. Anim. Sci. 44: Kubota, J., and W. H. Allaway Geographic distribution of trace element problems. Page 525 in Micronutrients in agriculture. J. J. Mortvedt, W. L. Lindsay, and P. M. Giordano, eds. Soil Sci. Am., Inc., Madison, WI. 14 Logan, E. E., A. Stenhouse, and D. J. Ormrod The role of colostral immunoglobulins in intestinal immunity to enteric colibacillosis in the calf. Res. Vet. Sci. 17: Moser, E. A., W. E. Julien, and D. L. Palmquist Response of neonatal calves to selenium supplementation. J. Dairy Sci. 60 (Suppl. 1):183. (Abstr.) 16 National Research Council Nutrient requirements of dairy cattle. Natl. Acad. Sci., Washington, DC. 17 Oh, S. H., R. A. Sunde, A. L. Pope, and W. G. Hoekstra Glutathione peroxidase response to selenium intake in lambs fed a formula yeast based artificial milk. J. Anim. Sci. 42: Oldfield, J. E The selenium story: Some reflections on the "moon-metal". New Zealand Vet. J. 22: Olson, O. E., I. Palmer, and E. Cary Modification of the official fluorometric method for selenium in plants. J. Assoc. Offic. Anal. Chem. 58~117~ 20 Paglia, D. E., and W. N. Valentine Studies on the quantitative and qualitative characterization of erythrocyte glutathlone peroxidase. J. Lab. Clin. Med. 40: Perry, T. W., D. M. Caldwell, and R. C. Peterson Selenium content of feeds and effect of dietary selenium on hair and blood serum. J. Dairy Sci. 59: Rotruck, J. T., A. L. Pope, H. E. Ganther, A. B. Swanson, D. G. Hafeman, and W. G. Hoekstra Selenium. Biochemical role as a component of glutathione peroxidase. Science 179: Rotruck, J. T., A. L. Pope, H. E. Ganther, and W. G. Hoekstra Prevention of oxidative damage to rat erythrocytes by dietary selenium. J. Nutr. 102: Scholz, R. W., and L. J. Hutchinson Distribution of glutathione peroxidase activity and selenium in the blood of dairy cows. Am. J. Vet. Res. 40: Scholz, R. W., D. A. Todhunter, and L. S. Cook Selenium content and glutathione peroxidase activity of young cattle fed supplemental whole milk diets. Am. J. Vet. Res. 42: Siddons, R. L., and C. F. Mills Glutathione peroxidase activity and erythrocyte stability in calves differing in selenium and vitamin E status. Br. J. Nutr. 46: Spallholz, J. E., J. L. Martin, M. L. Gerlach, and R. H. Heinzerburg Immunologic responses of mice fed diets supplemented with selenite selenium. Proc. Soc. Exp. Biol. Med. 143: Steel, R.G.D., and J. H. Torrie Principles and procedures of statistics. 2rid ed. McGraw-Hill, Inc., New York, NY.