- Email: [email protected]
Effect of Vitamin E Supplementation on Weight Gain, Immune Competence, and Disease Incidence in Barley-Fed Beef Cattle
Effect of Vitamin E Supplementation on Weight Gain, Immune Competence, and Disease Incidence in Barley-Fed Beef Cattle B. PEHRSON,' J. HAKKARAINEN,~ M. T ~ R N Q U I S T , ~ K. EDFORS,' and C. FOSSUM4 College of Veterinary Medicine Swedish University of Agricultural Sciences Uppsala, Sweden ABSTRACT
low vitamin E status rather than positive effects of additional vitamin in the diet. (Key words: vitamin E supplementation, immune status, weight gain, disease incidence)
The aim of the study was to investigate whether vitamin E supplements in larger amounts than recommended could reduce incidence of disease, improve immune competence, and increase rate of weight gain of conventionally barley-fed beef cattle. Mean daily intake of vitamin E by individual calves in the experimental group was 200 mg during the first 2 mo, 400 mg during the next 2 mo, and 600 mg during the rest of the period. Corresponding daily intakes of vitamin E for the control group were 50, 100, and 150 mg. Mean plasma vitamin E of the experimental group increased from .49 m a at the start of the trial to 2.03 mg/L at the end, but that of the control group was lower at the end (.36 m a ) than at the beginning (.53 m a ) . No significant differences were observed between the groups concerning incidence of disease or magnitude of lymphocyte stimulation. The results indicated that there was a surprisingly poor biological availability of the dietary vitamin Therefore, a comparison in reality was made between calves with inadequate and normal vitamin E status. The differences in daily BW gain and time to reach slaughter weight thus probably were effects of the
Abbreviation key: ARC = Agricultural Research Council, GSH-Px = glutathione peroxidase. INTRODUCTION
Received May 29, 1990. Accepted October 18, 1990. 'Experimental station, veterinary Institute, POB 234, S-532 23 Skara 2Department of Clinical Nutrition,POB 7023, S-750 07 UppSala. ' S ~ e d i f h farmer^' Meat Marketing ASSOChtiOIl, Animal Health Service, S-532 87 Skara %artment of Veterinary Microbiology, POB 582, S751 23 Uppsala. 1991 J Dairy Sci 74:1054-1059
It is difficult to quantify the daily requirement of young growing cattle for vitamin E, partly because of the influence of other dietary factors (23) and partly because of the lack of accepted criteria for what constitutes a normal vitamin E status. The requirement for vitamin E proposed by the Agricultural Research Council (ARC) (1) is slightly lower than that propOsed by Roche (22). However, it seems generally to be accepted that nutritional muscular dystrophy may occur in animals with serum vitamin E concentrations less than 1.0 to 1.5 mg/L (3, 13, 18). It was surprising, therefore, that Pehrson and Hakkaminen (17) found that many barleyfed beef cattle in Sweden had serum vitamin E concentrations below this range, even though their diet had been supplemented by the amount of vitamin E recommended by the ARC (1). In barley-fed beef production, large numbers of calves aged 3 to 7 wk are often purchased from many different herds and raised together in big units. They are offered free access to concentrates and restricted amounts of roughage in order to attain a maximal daily weight gain. The system encourages the spread of infectious diseases, particularly those affecting the digestive and respiratory tracts. A high incidence of such diseases has been recorded in Swedish barley-fed cattle (14). It is lmown that vitamin E can stimulate the immune defense mechanisms in laboratory animals (16, 24), and such beneficial effects
1054
VITAMIN E AND BEEF C A W PERF'ORMANCE
1055
also have been recorded in cattle (5.21). More- was offered an additional supply of vitamin E over, vitamin E has been reported to improve as a powder (2% dla-tmpheryl acetate in the daily weight gain and food conversion effi- glucose), which was mixed into the milk replacer or spread over the concentrates. This ciency of young cattle (7, 12, 19). This study was to investigate whether sup- extra dose of vitamin E was increased twice plements of more than the recommended during the trial so that the mean daily intake of amounts of vitamin E could reduce incidence of vitamin E by individual calves was calculated disease, improve immune competence, and in- to be 200 mg during the first 2 mo, 400 mg crease rate of BW gain of conventionally bar- during the next 2 mo, and 600 mg during the ley-fed beef cattle kept under commercial con- rest of the experimental period. The corresponding figures for the control calves (which ditions. received glucose as a placebo) were 50, 100, and 150 mg of vitamin E daily. The actual MATERIALS AND METHODS intake of vitamin E by the calves was estabTwenty-six Swedish Red and White and 12 lished from measurements of feed intake and Swedish Friesian male calves were used. After analysis of samples of feed for vitamin E. they had been purchased from at least 25 differThe trial was performed as a double blind ent farms at 3 to 7 wk of age, they were divided test, i.e., neither the fanner nor the responsible into experimental and control groups each con- researcher knew which animals were experitaining equal numbers of calves of similar mental and controls until all evaluations were weights and breeds. The mean BW of the ex- done. perimental group was 53.5 kg, and that of the The incidence of disease was evaluated first control group was 54.4 kg. All calves were by measuring the body temperature of all the offered a commercial milk replacer for 45 d calves three times a week for the first 4 wk, after arriving at the fann and had free access to second by recording all treatments with antibiconcentrates supplemented with vitamins, min- otic drugs throughout the trial, and third by erals, and trace elements (consisting of a com- examining the organs of the animals after mercial protein feed with 28.0% digestible CP slaughter. Febrile animals that looked depressed and homegrown oats and barley grain with or had a reduced appetite or diarrhea were 8.0% digestible CP). The proportions between treated with antibiotics. The daiIy weight gain of the calves was protein feed and grain were changed so that the level of digestible CP was reduced gradually measured by weighing them at the beginning of from 12.5% at the beginning to 9.0% at the end the trial and six times during the trial. The of the experiment. The daily allowance of hay calves were slaughtered when they reached a was .5 kg/calf, but straw was substituted for predetermined weight of about 420 kg (about hay during the last 2.5 mo. 240 kg for the light barley-fed beef calves). The concentration of vitamin E in blood During the first 5 mo, calves were kept four to a box. Six calves in the experimental group plasma was measured by the HPLC technique and five calves in the control group then were of Hakkarainen et al. (6) at the beginning of the slaughtered as light barley beef calves in accor- trial and after 6 and 8 mo. Blood samples were dance with the conventional space-saving pro- withdrawn from the jugular vein in heparinized cedure used in the herd The other calves were vacutainers and immediately centrifuged. moved to a new bam, where six to nine calves Plasma was frozen and stored at -20'C until were kept in each box. All the boxes had slatted analysed. The same HPLC technique was used floors. The feed consumption per box was reg- to measure vitamin E in all of the feedstuffs, istered once a month during periods of 4 d. The activity of the enzyme glutathione perIn addition to the natural vitamin E content oxidase (GSH-Px) in erythrocytes was meaof the diet, all the calves received extra vitamin sured four times during the trial by the method E (dl-a-tocophexyl acetate) in the milk replacer of Carlstrom et al. (4); this activity was used as and in the concentrates. The milk replacer was an indication of the Se status of the calves. supplemented with 60 mg vitamin E/kg of powA lymphocyte stimulation test was perder, and the concentrates were supplemented formed 5 mo after the beginning of the trial by with 10 to 20 mgflrg. The experimental group the method of Lamon et al. (11). Whole blood Journal of Dairy Science Vol. 74, No. 3, 1991
1056
PEHRSON ET AL.
TABLE 1. Plasma vitamin E (miuignrms of cxrtocophesol per liter) in 13 vitamin E-supplemented experimental and 14 unsupplemented control calves (calves slaughtered after 5 mo were excluded). ~~
At start
*UP
After5mo
X
SD Experimental Control Significance
.49 .53
.20 .19
SD -62
1.34 .29 .15 P < .001
NS1
After 7 mo
X
2.03 .36
SD .91 .14
P < .001
'(P > .05).
cultures were used with poke-weed mitogen (Boehringer, Mannheim, Germany) at 10 pg/ ml, Concanavalin A (Pharmacia, Uppsala, Sweden) at 5 pg/ml, or leucoagglutinin (Pharmacia, Uppsala, Sweden) at 5 pg/ml, as mitogens. The results are presented as logarithms of the net counts per minute (log net cpm), where net cpm was calculated as the gross cpm in mitogen-stimulated cultures minus cpm in the negative control cultures (a culture without mitogen). In order to increase the chance of detecting a difference in immune competence, control calves with a concentration of vitamin E in plasma less than .20 mg/L were compared with experimental calves with a plasma vitamin E concentration greater than 1.50 m a . Statistical evaluations were made by conventional ANOVA using P = .05.
the experimental calves it ranged from .34 to 3.83 m a ; 25%of the experimental calves had a vitamin E concentration less than 1.0 m&. a-Tocopherol was the dominant isomer of vitamin E in the plasma of all calves. Traces of ytocopherol occasionally were detected in the young calves while they were s t i l l eating milk replacer, and traces of a-tocotrienol more often were detected in the older barley-fed beef animals. No significant differences were observed between the activities of GSH-Px in the erythrocytes of the two groups of calves at any time. The mean (* SD) activity in the two groups considered together increased from 1151 f 354 pkatL at the beginning to 2031 f 138 pkat/L at the end of the trial. According to Carlstr6m et al. (4). these GSH-Px activities correspond to whole blood Se concentrations of 1 13 and 187 ng/ml, respectively, and indicate that the Se
RESULTS
The actual intake of vitamin E by control calves (Figure 1) was slightly less than that estimated on the basis of the assumed feed intakes. Nevertheless, it was very close to the requirement proposed by the ARC (1) although slightly less than that proposed by Roche (22). The intake of vitamin E by experimental calves was close to the calculated estimate except during the last 2.5 mo of the trial. It was four times greater than the vitamin E intake of the control calves and much greater than the highest of the proposed requirement values. The concentration of vitamin E (a-tocopher01) in plasma of the experimental calves inI . . 0 3 4 5 6 7 S u o creased during the trial, but the concentration in Figure 1. The vitamin E (WT. E) intake of the expenthe control calves was lower during the trial than at the beginning (Table 1). During the mental A) and control B) p u p s of calves from start of the trial in comparisonwith their requirements according to the trial, vitamin E concentrations in the control Agridtmal Research Council (ARC) (1) and Roche (22). calves ranged from .06 to .67 mg/L, whereas in All data have been transEormed to dl-a-tocopberyl acefate. 1
Journal of Dairy Science Vol. 74, No. 3, 1991
1
1057
VITAMIN E AND BEEF CATTLE PE3(poRMANcE
TABLE 2. Daily weight gain (gnuns) in vitamin E-supplemented experimental calves and in unsupplemented control calves during the k t 5 mo and during the last 4 mo of the trial. ~
n 19 19
Experimental Control Significance
'(P >
During the last 4 mo
huing the first 5 mo
Group
X
SD
n X 13 1241 12 1125 P < .05
1204 83 1147 47
NS'
SD 121 129
.os).
status of all the calves was normal throughout the trial. There was no significant difference between the incidence of disease in the two groups of calves. During the first few weeks, diarrhea was common among both groups. In the experimental group, 75% of the calves had an elevated temperature at some time during the first 4 wk (compared with 70% of the control calves), 85% were treated with antibiotics (compared with 70% of the control calves), and at slaughter 6 had liver abscesses (compared with 6 control calves), and 2 had pneumonia (compared with 3 control calves). Two of the control calves became seriously ill during the second part of the trial; they lost a great deal of body condition, owing to a subcutaneous abscess in one case and arthritis in the other. For this reason, they were excluded from the statistical comparisons of daily weight gain and time taken to reach slaughter weight. On average, the experimental calves gained weight more rapidly than the control calves throughout the trial; the difference was significant during the last 4 mo (Table 2). As a result, experimental calves reached slaughter weight in
a mean (fSD) time of 286 f 12 d, a sign& cantly (P< .001) shorter period than the 311 f 24 d required by controls. The carcass weights of the two groups were very similar, 219 f 6 vs. 215 f 9 kg for the experimental and control PUPS. No significant differences were observed with any of the three mitogens employed between the magnitudes of lymphocyte stimulation for control calves with very low plasma concentrations of vitamin E and experimental calves with high concentrations of vitamin E (Table 3). DISCUSSION
This investigation provides further evidence that many barley-fed beef cattle in Sweden have remarkably low blood concentrations of vitamin E (17). The highest vitamin E concentration in a control calf was only .67 mg/L, in spite of the fact that the calves' daily intake of vitamin E was close to that recommended by the ARC (1). The mean plasma vitamin E concentrations in the experimental calves (1.34 mgL after 5 mo and 2.03 m& after 7 mo on
TABLE 3. Responses of lymphocytes to stimulation by three different mitogens, expressed as the logaxithms of the net counts per minute (log net cpm)in 8 vitamin E-supplrmented experimental calves with reasonably high serum vitamin E Concentrations (> 1.50 mg/L) and in 8 nonsupplemented conbol calves with very low serum vitamin E concentrations (< .20 mg/L).
Serum Grouu
X Experimental Control Significance
Lcucoanalutinin
vitamin E
(mg/L)
SD
1.96 .36 .13 .05 P < .001
--
SD
4.25
.59 .61
NS1
concanavalin ~~~~
~~~
Oog net cpm)
X
4-53
Poke weed
X
523 5.16 NS
SD
SD .10 .07
5.04 5.08
.14 .16
NS1
'(P > .05). Journal of Dairy Science Vol. 74, No. 3, 1991
1058
PEHRSON ET AL.
the supplemented diet) also were remarkably low in relation to their high intake of vitamin E. There also was a very wide range (.34 to 3.83 mg/L) of vitamin E concentrations among the experimental calves, and 25% of them had plasma concentrations less than 1.0 mg/L, a value below which calves may develop nub-itional muscular dystrophy (3, 13, 18). The apparent poor biological availability of supplementary dietary vitamin E and the wide variability in response of individual animals to supplementation may be due to degradation of the vitamin by rumen microorganisms in some animals fed a high grain diet, as has earlier been found by Alderson et al. (2). Another possibility could be a lower than expected digestibility of the vitamin in ruminants, as reported by Hidiroglou and Karpinski (8). It has also earlier been reported that tocopherol acetate might be utilized less efficiently than free tocopherol (9). Whatever the reasons for the poor response to vitamin E supplementation, the vitamin E requirements proposed by ARC (1) are likely too low for barley-fed cattle, as indicated also by the results presented by Reddy et al. (19). Barley beef production in Sweden suffers from a high incidence of infectious disease, principally because of the way in which calves from many herds are mixed together. The incidence of disease during t h i s trial proved to be no exception, and it was particularly high during the first 4 wk ‘Ihe fact that the additional vitamin E provided to the experimental group of calves did not reduce the incidence of disease during this period could have been b e cause the infectious agent attacked the calved only a few days after they had started the supplemented diet, before any immunostimulatory effect of the vitamin E could have deve loped. However, even after 5 mo on the diet, the in vitro lymphocyte stimulation tests gave no indication of any increase in the immune status of the supplemented calves. In most cases, the immunostimulatoxy effects of additional vitamin E have been reported to be associated with supplementation in excess of levels determined to meet the recommended minimum requirements (15, 25). Although very large doses of vitamin E were administered to the experimental calves, their plasma vitamin E concentrations were not markedly increased, and it is therefore not surJournal of Dairy Science Vol. 74, No. 3, 1991
prising that an immunostimulatory effect was not observed. It would be valuable to investigate whether large parenteral doses of vitamin E might achieve sufficiently large increases in plasma vitamin E concentrations to stimulate the calves’ immune system and thus help protect them against virulent strains of infectious agents during the first few weeks after purchase. Reddy et al. (20, 21) found that the parenteral administration to calves of 1400 mg vitamin E once a week increased their serum vitamin E concentrations to 3.4 to 4.8mg/L and significantly increased their in vitro lymphocyte stimulation indices. Furthermore, Jensen et al. (10) found that in growing pigs a serum vitamin E concentration above 3 mg/L was necessary to achieve a signrficant response of the lymphocytes to stimulation with mitogens. The original aim of this study was to compare groups of barley-fed beef cattle that had either a high or a normal vitamin E status. Owing to the unexpectedly poor biological availability of the dietary vitamin, a comparison has been made between groups of calves that had either an inadequate or a reasonably normal vitamin E status. Therefore, the differences between the groups, in terms of daily weight gain and time to reach slaughter weight, were probably due to the negative effects of the low vitamin E status of the control calved rather than to the positive effects of additional vitamin E in the diet of the experimental calves. Further experiments are needed to elucidate the mechanisms responsible for the apparent poor availability of vitamin E fed to calves. REFERENCES 1 Agricultural Research Council. 1982. The nutrient requirements of nuninant livestock. Commonwealth Agricultural Bureau, Slough, Engl. 2 Alderson, N. E., G. E. Mitchell Jr., C. 0. Little, R. E. Warner, and R. E. Tucker. 1971. Preintestinal disap pearance of vitamin E in nlminnnb. J. Nutr. 101:21. 3Arthur, J. N. 1982. N u h i t b ~ linterrelati~nshipbetween selenium and vitamin E. Annu. Rep. No. 38124, Rowett Res. Inst., Aberdeen, Scotland. 4CarktrBm. G., G. J b s o n , and B. Pehrson. 1979. An evaluation of selenium status of cattle in Sweden by means of glutathione peroxidase. Swed. J. Agric. Res. 9:43. 5Ciprian0, J. E., J. L. Monill. and N. V. Anderson 1982. Effect of dietary vitamin E on immune responses of dairy calves. I. Dairy Sci. 65:2357. 6Hakkamm ’ n, R.V.J.,J. T. Tyapptinen, S. Hassari, S. G. Bagtsson, S.R.L.. Jawon, and P. 0. Lindberg. 1984. Biopotency of vitamin E io barley. Br. J. Nu&. 52335.
VITAMIN E AND BEEF CA’lTLE PERFORMANCE
7Hicks, R. B. 1985. Effect of nutrition, medical treatments and management practices on health and performance of newly received stocker cattle. M S. Thesis, Oklahoma State Univ.. Stillwater. 8Hidiroglou, M., and K. Karpinski. 1987. Vitamin E kinetics in sheep. Br. J. Nutr. 58:113. 9Hidiroglou, M., L. R McDowell, and 0.Balbuena. 1989. Plasma tocopherol in sheep and cattle after ingesting free or acetylated tocopherol. J. Dairy Sci. 72: 1793. 10 Jensen, M., C. Fossum, M.Ederoth, and R.VJ. Hakkarainen. 1988. The effect of vitamin E on the cellmediated immune response in pigs. I. Vet. Med. 35: 549. 11 Larsson, B., C. Fossum, and S. Alenius. 1988. A cellular analysis of immunosuppression in cattle with mucosal disease. Res. Vet. Sci. 44:71. 12Lee, R. W.,R L. Stuart. K.R. Pemyman, and K.W. Ridenour. 1985. Eflect of vitamin supplementation on the performance of stressed beef calves. J. Anim. Sci. 61~425. 13 McMurray, C. H., and D. A. Rice. 1982. Vitamin E and selmiUm deficiency diseases. Ir. Vel. J. 3657. 14Moreno-Lopez, J., and M. T6rnquist. 1980. Vaccination against acute respiratory/enteric disease during calfhood. Page 388 in 11th Int. Congr. Dis. Cattle, Tel Aviv, 1-1. lSNafstad, I. 1983. Current knowledge of vitamin E. Biochemical functions and siguificance in diseases. Page 5 in 1982 Publications of the Norwegian College of Veterinary Medicine, Oslo, Norway. 16 Nockels. C. F. 1979. Protective effects of supplemental vitamin E against infection. Fed. Roc. 38:2134.
1059
17Pehrsor1, B., and J. Hakkamn ’ en. 1986. Vitamin E status of healthy Swedish cattle. Acta Vet. Scand. 27: 351. 18Pehrson, B.,J. H & ~ E ’I Un,E and J. Ty6pp6nen. 1986. Nutritional muscular degeneration in young heifers. Nord. Vet. Med. 38:26. 19Reddy, P. G., J. L. Monill, and R A. Prey. 1987. Vitamin E requirements of d;ury calves. J. Dairy Sci. 70:123. 20 Reddy, P. G., I. L. Morrill, R.A. Frey,M. B. Morrill, H. C. Minocha, S. J. Galiker,and A. D.Dayton. 1985. Effects of supplemental vitamin E on the performance and metabolic profilcs of dairy calves. I. Dairy Sci. 6 8 2259. 21Reddy, P. G., J. L. M o d , H. C. Minocha, M. B. Morrill, A. D. Dayton, and R A. Frey. 1986. Effects of supplemental vitamin E on the immune system of calves. J. Dairy Sci. 69164. 22 Roche. 1987. Recommended vitamin supplementation levels for domestic animals.F’amphlek F. Hoffman LaRoche & Co AG, Basel, Switimhnd. 23Scott, M. L. 1978. Vitamin E. Page 133 in The fat soluble vitamins. Handbook of lipid research. 2nd ed. H. E. Dehca, ed Plenum Press, New York and
London. 2 4 S h e f f y , B. E., and R D. Schnltz. 1979. Influence of vitamin E and selenium on immune response mechanisms. Fed. Proc. 38:2139. 25 Tengerdy, R P., M M.Mathias, and C. F. Nockels. 1981. Vitamin E, immunity and disease resistance. Page 27 in Diet and resistance to disease. M Phillips and A. Baek, ed. Plenum Press, New York and London.
Journal of Daiq Science Vol. 74, No. 3, 1991
low vitamin E status rather than positive effects of additional vitamin in the diet. (Key words: vitamin E supplementation, immune status, weight gain, disease incidence)
The aim of the study was to investigate whether vitamin E supplements in larger amounts than recommended could reduce incidence of disease, improve immune competence, and increase rate of weight gain of conventionally barley-fed beef cattle. Mean daily intake of vitamin E by individual calves in the experimental group was 200 mg during the first 2 mo, 400 mg during the next 2 mo, and 600 mg during the rest of the period. Corresponding daily intakes of vitamin E for the control group were 50, 100, and 150 mg. Mean plasma vitamin E of the experimental group increased from .49 m a at the start of the trial to 2.03 mg/L at the end, but that of the control group was lower at the end (.36 m a ) than at the beginning (.53 m a ) . No significant differences were observed between the groups concerning incidence of disease or magnitude of lymphocyte stimulation. The results indicated that there was a surprisingly poor biological availability of the dietary vitamin Therefore, a comparison in reality was made between calves with inadequate and normal vitamin E status. The differences in daily BW gain and time to reach slaughter weight thus probably were effects of the
Abbreviation key: ARC = Agricultural Research Council, GSH-Px = glutathione peroxidase. INTRODUCTION
Received May 29, 1990. Accepted October 18, 1990. 'Experimental station, veterinary Institute, POB 234, S-532 23 Skara 2Department of Clinical Nutrition,POB 7023, S-750 07 UppSala. ' S ~ e d i f h farmer^' Meat Marketing ASSOChtiOIl, Animal Health Service, S-532 87 Skara %artment of Veterinary Microbiology, POB 582, S751 23 Uppsala. 1991 J Dairy Sci 74:1054-1059
It is difficult to quantify the daily requirement of young growing cattle for vitamin E, partly because of the influence of other dietary factors (23) and partly because of the lack of accepted criteria for what constitutes a normal vitamin E status. The requirement for vitamin E proposed by the Agricultural Research Council (ARC) (1) is slightly lower than that propOsed by Roche (22). However, it seems generally to be accepted that nutritional muscular dystrophy may occur in animals with serum vitamin E concentrations less than 1.0 to 1.5 mg/L (3, 13, 18). It was surprising, therefore, that Pehrson and Hakkaminen (17) found that many barleyfed beef cattle in Sweden had serum vitamin E concentrations below this range, even though their diet had been supplemented by the amount of vitamin E recommended by the ARC (1). In barley-fed beef production, large numbers of calves aged 3 to 7 wk are often purchased from many different herds and raised together in big units. They are offered free access to concentrates and restricted amounts of roughage in order to attain a maximal daily weight gain. The system encourages the spread of infectious diseases, particularly those affecting the digestive and respiratory tracts. A high incidence of such diseases has been recorded in Swedish barley-fed cattle (14). It is lmown that vitamin E can stimulate the immune defense mechanisms in laboratory animals (16, 24), and such beneficial effects
1054
VITAMIN E AND BEEF C A W PERF'ORMANCE
1055
also have been recorded in cattle (5.21). More- was offered an additional supply of vitamin E over, vitamin E has been reported to improve as a powder (2% dla-tmpheryl acetate in the daily weight gain and food conversion effi- glucose), which was mixed into the milk replacer or spread over the concentrates. This ciency of young cattle (7, 12, 19). This study was to investigate whether sup- extra dose of vitamin E was increased twice plements of more than the recommended during the trial so that the mean daily intake of amounts of vitamin E could reduce incidence of vitamin E by individual calves was calculated disease, improve immune competence, and in- to be 200 mg during the first 2 mo, 400 mg crease rate of BW gain of conventionally bar- during the next 2 mo, and 600 mg during the ley-fed beef cattle kept under commercial con- rest of the experimental period. The corresponding figures for the control calves (which ditions. received glucose as a placebo) were 50, 100, and 150 mg of vitamin E daily. The actual MATERIALS AND METHODS intake of vitamin E by the calves was estabTwenty-six Swedish Red and White and 12 lished from measurements of feed intake and Swedish Friesian male calves were used. After analysis of samples of feed for vitamin E. they had been purchased from at least 25 differThe trial was performed as a double blind ent farms at 3 to 7 wk of age, they were divided test, i.e., neither the fanner nor the responsible into experimental and control groups each con- researcher knew which animals were experitaining equal numbers of calves of similar mental and controls until all evaluations were weights and breeds. The mean BW of the ex- done. perimental group was 53.5 kg, and that of the The incidence of disease was evaluated first control group was 54.4 kg. All calves were by measuring the body temperature of all the offered a commercial milk replacer for 45 d calves three times a week for the first 4 wk, after arriving at the fann and had free access to second by recording all treatments with antibiconcentrates supplemented with vitamins, min- otic drugs throughout the trial, and third by erals, and trace elements (consisting of a com- examining the organs of the animals after mercial protein feed with 28.0% digestible CP slaughter. Febrile animals that looked depressed and homegrown oats and barley grain with or had a reduced appetite or diarrhea were 8.0% digestible CP). The proportions between treated with antibiotics. The daiIy weight gain of the calves was protein feed and grain were changed so that the level of digestible CP was reduced gradually measured by weighing them at the beginning of from 12.5% at the beginning to 9.0% at the end the trial and six times during the trial. The of the experiment. The daily allowance of hay calves were slaughtered when they reached a was .5 kg/calf, but straw was substituted for predetermined weight of about 420 kg (about hay during the last 2.5 mo. 240 kg for the light barley-fed beef calves). The concentration of vitamin E in blood During the first 5 mo, calves were kept four to a box. Six calves in the experimental group plasma was measured by the HPLC technique and five calves in the control group then were of Hakkarainen et al. (6) at the beginning of the slaughtered as light barley beef calves in accor- trial and after 6 and 8 mo. Blood samples were dance with the conventional space-saving pro- withdrawn from the jugular vein in heparinized cedure used in the herd The other calves were vacutainers and immediately centrifuged. moved to a new bam, where six to nine calves Plasma was frozen and stored at -20'C until were kept in each box. All the boxes had slatted analysed. The same HPLC technique was used floors. The feed consumption per box was reg- to measure vitamin E in all of the feedstuffs, istered once a month during periods of 4 d. The activity of the enzyme glutathione perIn addition to the natural vitamin E content oxidase (GSH-Px) in erythrocytes was meaof the diet, all the calves received extra vitamin sured four times during the trial by the method E (dl-a-tocophexyl acetate) in the milk replacer of Carlstrom et al. (4); this activity was used as and in the concentrates. The milk replacer was an indication of the Se status of the calves. supplemented with 60 mg vitamin E/kg of powA lymphocyte stimulation test was perder, and the concentrates were supplemented formed 5 mo after the beginning of the trial by with 10 to 20 mgflrg. The experimental group the method of Lamon et al. (11). Whole blood Journal of Dairy Science Vol. 74, No. 3, 1991
1056
PEHRSON ET AL.
TABLE 1. Plasma vitamin E (miuignrms of cxrtocophesol per liter) in 13 vitamin E-supplemented experimental and 14 unsupplemented control calves (calves slaughtered after 5 mo were excluded). ~~
At start
*UP
After5mo
X
SD Experimental Control Significance
.49 .53
.20 .19
SD -62
1.34 .29 .15 P < .001
NS1
After 7 mo
X
2.03 .36
SD .91 .14
P < .001
'(P > .05).
cultures were used with poke-weed mitogen (Boehringer, Mannheim, Germany) at 10 pg/ ml, Concanavalin A (Pharmacia, Uppsala, Sweden) at 5 pg/ml, or leucoagglutinin (Pharmacia, Uppsala, Sweden) at 5 pg/ml, as mitogens. The results are presented as logarithms of the net counts per minute (log net cpm), where net cpm was calculated as the gross cpm in mitogen-stimulated cultures minus cpm in the negative control cultures (a culture without mitogen). In order to increase the chance of detecting a difference in immune competence, control calves with a concentration of vitamin E in plasma less than .20 mg/L were compared with experimental calves with a plasma vitamin E concentration greater than 1.50 m a . Statistical evaluations were made by conventional ANOVA using P = .05.
the experimental calves it ranged from .34 to 3.83 m a ; 25%of the experimental calves had a vitamin E concentration less than 1.0 m&. a-Tocopherol was the dominant isomer of vitamin E in the plasma of all calves. Traces of ytocopherol occasionally were detected in the young calves while they were s t i l l eating milk replacer, and traces of a-tocotrienol more often were detected in the older barley-fed beef animals. No significant differences were observed between the activities of GSH-Px in the erythrocytes of the two groups of calves at any time. The mean (* SD) activity in the two groups considered together increased from 1151 f 354 pkatL at the beginning to 2031 f 138 pkat/L at the end of the trial. According to Carlstr6m et al. (4). these GSH-Px activities correspond to whole blood Se concentrations of 1 13 and 187 ng/ml, respectively, and indicate that the Se
RESULTS
The actual intake of vitamin E by control calves (Figure 1) was slightly less than that estimated on the basis of the assumed feed intakes. Nevertheless, it was very close to the requirement proposed by the ARC (1) although slightly less than that proposed by Roche (22). The intake of vitamin E by experimental calves was close to the calculated estimate except during the last 2.5 mo of the trial. It was four times greater than the vitamin E intake of the control calves and much greater than the highest of the proposed requirement values. The concentration of vitamin E (a-tocopher01) in plasma of the experimental calves inI . . 0 3 4 5 6 7 S u o creased during the trial, but the concentration in Figure 1. The vitamin E (WT. E) intake of the expenthe control calves was lower during the trial than at the beginning (Table 1). During the mental A) and control B) p u p s of calves from start of the trial in comparisonwith their requirements according to the trial, vitamin E concentrations in the control Agridtmal Research Council (ARC) (1) and Roche (22). calves ranged from .06 to .67 mg/L, whereas in All data have been transEormed to dl-a-tocopberyl acefate. 1
Journal of Dairy Science Vol. 74, No. 3, 1991
1
1057
VITAMIN E AND BEEF CATTLE PE3(poRMANcE
TABLE 2. Daily weight gain (gnuns) in vitamin E-supplemented experimental calves and in unsupplemented control calves during the k t 5 mo and during the last 4 mo of the trial. ~
n 19 19
Experimental Control Significance
'(P >
During the last 4 mo
huing the first 5 mo
Group
X
SD
n X 13 1241 12 1125 P < .05
1204 83 1147 47
NS'
SD 121 129
.os).
status of all the calves was normal throughout the trial. There was no significant difference between the incidence of disease in the two groups of calves. During the first few weeks, diarrhea was common among both groups. In the experimental group, 75% of the calves had an elevated temperature at some time during the first 4 wk (compared with 70% of the control calves), 85% were treated with antibiotics (compared with 70% of the control calves), and at slaughter 6 had liver abscesses (compared with 6 control calves), and 2 had pneumonia (compared with 3 control calves). Two of the control calves became seriously ill during the second part of the trial; they lost a great deal of body condition, owing to a subcutaneous abscess in one case and arthritis in the other. For this reason, they were excluded from the statistical comparisons of daily weight gain and time taken to reach slaughter weight. On average, the experimental calves gained weight more rapidly than the control calves throughout the trial; the difference was significant during the last 4 mo (Table 2). As a result, experimental calves reached slaughter weight in
a mean (fSD) time of 286 f 12 d, a sign& cantly (P< .001) shorter period than the 311 f 24 d required by controls. The carcass weights of the two groups were very similar, 219 f 6 vs. 215 f 9 kg for the experimental and control PUPS. No significant differences were observed with any of the three mitogens employed between the magnitudes of lymphocyte stimulation for control calves with very low plasma concentrations of vitamin E and experimental calves with high concentrations of vitamin E (Table 3). DISCUSSION
This investigation provides further evidence that many barley-fed beef cattle in Sweden have remarkably low blood concentrations of vitamin E (17). The highest vitamin E concentration in a control calf was only .67 mg/L, in spite of the fact that the calves' daily intake of vitamin E was close to that recommended by the ARC (1). The mean plasma vitamin E concentrations in the experimental calves (1.34 mgL after 5 mo and 2.03 m& after 7 mo on
TABLE 3. Responses of lymphocytes to stimulation by three different mitogens, expressed as the logaxithms of the net counts per minute (log net cpm)in 8 vitamin E-supplrmented experimental calves with reasonably high serum vitamin E Concentrations (> 1.50 mg/L) and in 8 nonsupplemented conbol calves with very low serum vitamin E concentrations (< .20 mg/L).
Serum Grouu
X Experimental Control Significance
Lcucoanalutinin
vitamin E
(mg/L)
SD
1.96 .36 .13 .05 P < .001
--
SD
4.25
.59 .61
NS1
concanavalin ~~~~
~~~
Oog net cpm)
X
4-53
Poke weed
X
523 5.16 NS
SD
SD .10 .07
5.04 5.08
.14 .16
NS1
'(P > .05). Journal of Dairy Science Vol. 74, No. 3, 1991
1058
PEHRSON ET AL.
the supplemented diet) also were remarkably low in relation to their high intake of vitamin E. There also was a very wide range (.34 to 3.83 mg/L) of vitamin E concentrations among the experimental calves, and 25% of them had plasma concentrations less than 1.0 mg/L, a value below which calves may develop nub-itional muscular dystrophy (3, 13, 18). The apparent poor biological availability of supplementary dietary vitamin E and the wide variability in response of individual animals to supplementation may be due to degradation of the vitamin by rumen microorganisms in some animals fed a high grain diet, as has earlier been found by Alderson et al. (2). Another possibility could be a lower than expected digestibility of the vitamin in ruminants, as reported by Hidiroglou and Karpinski (8). It has also earlier been reported that tocopherol acetate might be utilized less efficiently than free tocopherol (9). Whatever the reasons for the poor response to vitamin E supplementation, the vitamin E requirements proposed by ARC (1) are likely too low for barley-fed cattle, as indicated also by the results presented by Reddy et al. (19). Barley beef production in Sweden suffers from a high incidence of infectious disease, principally because of the way in which calves from many herds are mixed together. The incidence of disease during t h i s trial proved to be no exception, and it was particularly high during the first 4 wk ‘Ihe fact that the additional vitamin E provided to the experimental group of calves did not reduce the incidence of disease during this period could have been b e cause the infectious agent attacked the calved only a few days after they had started the supplemented diet, before any immunostimulatory effect of the vitamin E could have deve loped. However, even after 5 mo on the diet, the in vitro lymphocyte stimulation tests gave no indication of any increase in the immune status of the supplemented calves. In most cases, the immunostimulatoxy effects of additional vitamin E have been reported to be associated with supplementation in excess of levels determined to meet the recommended minimum requirements (15, 25). Although very large doses of vitamin E were administered to the experimental calves, their plasma vitamin E concentrations were not markedly increased, and it is therefore not surJournal of Dairy Science Vol. 74, No. 3, 1991
prising that an immunostimulatory effect was not observed. It would be valuable to investigate whether large parenteral doses of vitamin E might achieve sufficiently large increases in plasma vitamin E concentrations to stimulate the calves’ immune system and thus help protect them against virulent strains of infectious agents during the first few weeks after purchase. Reddy et al. (20, 21) found that the parenteral administration to calves of 1400 mg vitamin E once a week increased their serum vitamin E concentrations to 3.4 to 4.8mg/L and significantly increased their in vitro lymphocyte stimulation indices. Furthermore, Jensen et al. (10) found that in growing pigs a serum vitamin E concentration above 3 mg/L was necessary to achieve a signrficant response of the lymphocytes to stimulation with mitogens. The original aim of this study was to compare groups of barley-fed beef cattle that had either a high or a normal vitamin E status. Owing to the unexpectedly poor biological availability of the dietary vitamin, a comparison has been made between groups of calves that had either an inadequate or a reasonably normal vitamin E status. Therefore, the differences between the groups, in terms of daily weight gain and time to reach slaughter weight, were probably due to the negative effects of the low vitamin E status of the control calved rather than to the positive effects of additional vitamin E in the diet of the experimental calves. Further experiments are needed to elucidate the mechanisms responsible for the apparent poor availability of vitamin E fed to calves. REFERENCES 1 Agricultural Research Council. 1982. The nutrient requirements of nuninant livestock. Commonwealth Agricultural Bureau, Slough, Engl. 2 Alderson, N. E., G. E. Mitchell Jr., C. 0. Little, R. E. Warner, and R. E. Tucker. 1971. Preintestinal disap pearance of vitamin E in nlminnnb. J. Nutr. 101:21. 3Arthur, J. N. 1982. N u h i t b ~ linterrelati~nshipbetween selenium and vitamin E. Annu. Rep. No. 38124, Rowett Res. Inst., Aberdeen, Scotland. 4CarktrBm. G., G. J b s o n , and B. Pehrson. 1979. An evaluation of selenium status of cattle in Sweden by means of glutathione peroxidase. Swed. J. Agric. Res. 9:43. 5Ciprian0, J. E., J. L. Monill. and N. V. Anderson 1982. Effect of dietary vitamin E on immune responses of dairy calves. I. Dairy Sci. 65:2357. 6Hakkamm ’ n, R.V.J.,J. T. Tyapptinen, S. Hassari, S. G. Bagtsson, S.R.L.. Jawon, and P. 0. Lindberg. 1984. Biopotency of vitamin E io barley. Br. J. Nu&. 52335.
VITAMIN E AND BEEF CA’lTLE PERFORMANCE
7Hicks, R. B. 1985. Effect of nutrition, medical treatments and management practices on health and performance of newly received stocker cattle. M S. Thesis, Oklahoma State Univ.. Stillwater. 8Hidiroglou, M., and K. Karpinski. 1987. Vitamin E kinetics in sheep. Br. J. Nutr. 58:113. 9Hidiroglou, M., L. R McDowell, and 0.Balbuena. 1989. Plasma tocopherol in sheep and cattle after ingesting free or acetylated tocopherol. J. Dairy Sci. 72: 1793. 10 Jensen, M., C. Fossum, M.Ederoth, and R.VJ. Hakkarainen. 1988. The effect of vitamin E on the cellmediated immune response in pigs. I. Vet. Med. 35: 549. 11 Larsson, B., C. Fossum, and S. Alenius. 1988. A cellular analysis of immunosuppression in cattle with mucosal disease. Res. Vet. Sci. 44:71. 12Lee, R. W.,R L. Stuart. K.R. Pemyman, and K.W. Ridenour. 1985. Eflect of vitamin supplementation on the performance of stressed beef calves. J. Anim. Sci. 61~425. 13 McMurray, C. H., and D. A. Rice. 1982. Vitamin E and selmiUm deficiency diseases. Ir. Vel. J. 3657. 14Moreno-Lopez, J., and M. T6rnquist. 1980. Vaccination against acute respiratory/enteric disease during calfhood. Page 388 in 11th Int. Congr. Dis. Cattle, Tel Aviv, 1-1. lSNafstad, I. 1983. Current knowledge of vitamin E. Biochemical functions and siguificance in diseases. Page 5 in 1982 Publications of the Norwegian College of Veterinary Medicine, Oslo, Norway. 16 Nockels. C. F. 1979. Protective effects of supplemental vitamin E against infection. Fed. Roc. 38:2134.
1059
17Pehrsor1, B., and J. Hakkamn ’ en. 1986. Vitamin E status of healthy Swedish cattle. Acta Vet. Scand. 27: 351. 18Pehrson, B.,J. H & ~ E ’I Un,E and J. Ty6pp6nen. 1986. Nutritional muscular degeneration in young heifers. Nord. Vet. Med. 38:26. 19Reddy, P. G., J. L. Monill, and R A. Prey. 1987. Vitamin E requirements of d;ury calves. J. Dairy Sci. 70:123. 20 Reddy, P. G., I. L. Morrill, R.A. Frey,M. B. Morrill, H. C. Minocha, S. J. Galiker,and A. D.Dayton. 1985. Effects of supplemental vitamin E on the performance and metabolic profilcs of dairy calves. I. Dairy Sci. 6 8 2259. 21Reddy, P. G., J. L. M o d , H. C. Minocha, M. B. Morrill, A. D. Dayton, and R A. Frey. 1986. Effects of supplemental vitamin E on the immune system of calves. J. Dairy Sci. 69164. 22 Roche. 1987. Recommended vitamin supplementation levels for domestic animals.F’amphlek F. Hoffman LaRoche & Co AG, Basel, Switimhnd. 23Scott, M. L. 1978. Vitamin E. Page 133 in The fat soluble vitamins. Handbook of lipid research. 2nd ed. H. E. Dehca, ed Plenum Press, New York and
London. 2 4 S h e f f y , B. E., and R D. Schnltz. 1979. Influence of vitamin E and selenium on immune response mechanisms. Fed. Proc. 38:2139. 25 Tengerdy, R P., M M.Mathias, and C. F. Nockels. 1981. Vitamin E, immunity and disease resistance. Page 27 in Diet and resistance to disease. M Phillips and A. Baek, ed. Plenum Press, New York and London.
Journal of Daiq Science Vol. 74, No. 3, 1991