Effeds of Zinc, Copper,and Manganese Supplementation of High-Concentrate Ration on Digestibility, Growth, and Tissue Content of Holstein Calves M. IVAN 1 and C. M. GRIEVE Department of Animal Science, University of Alberta Edmonton, Alberta, Canada T6G 2E3 Abstract
Deficiencies of Zn (4, 17), Cu (11, 19), and Mn (3) decreased growth rate, which was improved by addition of the deficient mineral to the diet. Studies of effects of these individual trace minerals on performance of animals axe of little value because of interactions between the minerals. The main objective of our experiment was to study effects of addition of three trace minerals (Zn, Cu, and Mn) to a practical highconcentrate ration on the performance of bull calves. A second objective was to study interactions among these trace minerals. For all diets, measurements were of growth rate, efficiency of feed utilization, digestibility of dry matter (DM), nitrogen (N) and gross energy (GE), nitrogen retention (NR), excretion of Zn, Cu, and Mn in urine, and concentrations of minerals in liver, heart, and kidney tissues.
The effects of supplemental zinc, copper, and manganese alone or in combination in a high-concentrate ration were studied in 16 Holstein bull calves during a 10-wk feeding trial. Metabolism was studied after the feeding trial. Apparent digestibility of dry matter, nitrogen, and gross energy, and nitrogen retention and urinary excretion of zinc, copper, and manganese were determined. The calves were slaughtered after the experiment, and liver, heart, mad kidney were taken for analyses of trace minerals. Supplementation o~ the basal ration with the trace minerals did not affect body weight gains, which averaged 1.42 kg daily. The addition of trace minerals did not affect apparent digestibility coefficients. A zinc-manganese interaction in digestion of nitrogen and gross energy was significant. Higher dietary manganese caused increased zinc concentrations in the liver, kidney, and heart. The copper coneentratien of liver was decreased by dietary zinc and increased by dietary copper and manganese. Supplemental manganese increased its net retention. There was no evidence of deficiency of any trace minerals in the unsupplemented treatments.
Materials and Methods
Introduction Trace minerals are usually added to cattle rations to prevent or overcome deficiencies and
possible consequences on animal performance. The need for trace mineral supplementation has not been defined because of wide variation between feeds in their mineral content. This variation is due, in part at least, to the amount of elements in the soil, availability o~ soil minerals to the plants grown, and availability of plant minerals to animals fed. ,Received September 3, 1974. a Presemt address: Animal Research Institute,
Agriculture Canada, Ottawa, Ontario, Canada K1A 0(26.
Sixteen Holstein bull calves were allotted by weight in a 2 X 2 X 2 factorial design to eight treatments; (1) basal ration, (2) basal + Zn, (3) basal + Cu, (4) basal q- Zn _i_ Cu, (5) basal + Mn, (6) basal -t- Zn -t- Mn, (7) basal q- Cu + Mn, and (8) basal -b Zn q- (3u -k Mn. Calves in each treatment varied in average initial age from 22 to 28 wk and in average initial liveweight from 111 to 168 kg. They were in individual stalls and fed a d libitum. Water and a mixture of equal weights of calcium phosphate and eobaltized-iodized salt were available free choice. Ini6al and final individual liveweights were from two successive weighings at 24-h intervals after the calves were without feed and water overnight. The basal ration (Table 1) was formulated for all eight treatments and consisted primarily of coarsely ground barley. Urea was ineluded to provide a calculated 12% of protein equivalent. Experimental rations (Table 2) were formulated by addition of salts of particular trace minerals to the basal ration. Trace mineral salts were added to provide I00 ppm Zn, 20 ppm Cu, and 50 ppm Mn in the ration. Barley con-
410
411
TRACE MIN'E,RALS FOR CALVES
ing the period of metabolism study. A 5% aliquot was collected twice daily in a polyethylene bottle and stored at 4 G for chemical analysis. The calves were slaughtered the day after metabolism studies were completed. The heart, right kidney, and samples o f liver from each calf were taken. These samples were dried in a freeze-drier for 3 days, then homogenized in a laboratory blender and stored for analysis of trace minerals. The methods of AOAC (2) were used for determination of DM in the feed and fecal samples and of N in the feed, fecal, and urine samples. Gross energy in feed and fecal samples was measured by combustion in a Parr oxygen bomb calorimeter. Determination of Cr~Os in the feed and fecal samples was according to the method of Hill and Anderson (8). Atomic Absorption Spectrophotometry (Techtror~ AA-3) was used for analyses of Cu, Zn, and Mn. Samples of feed, liver, kidney, heart, and freeze-dried urine were prepared by wet ashing as outlined in AOAC (2). Statistical analyses were on the IBM 360/ 67 computer by a program written by Smillie
TABLE 1. Composition of the basal ration. ~e~e~s
%
Barley Urea Limestone Cobaltized-Iodized salt Vitamin A (10,000 IU/g) Vitamin D (35,000 IU/g) Vitamin E (44 IU/g) Chemical analysis Dry matter (%) Crude protein (~) Gross energy (Meal/kg) Calcium (%) Phosphorus (%)
97.6 .8 1.0 .5 .088 .0055 .015 88.1 12.6 4.12 .46 .40
tained 40 ppm, 6 ppm, and 12 ppm of Zn, Cu, and Mn. Metabolism was studied after the 10-wk feeding was completed. Chromic oxide (Cr2Os) was the indicator of fecal excretion (12). Each ration fed to experimental animals had .5% of Cr2Os mixed into it. To ensure complete consumption of the daily ration, feed offered was restricted by .5 kg below the average daily consumption for each calf during the final week of the feeding period. Daily rations were divided into two parts and fed twice daily for 12 days. A 5-g sample of feed was taken at each feeding. This was composited to form one sample per each ration, ground, and stored for analysis. Fecal grab-samples were obtained four times daily, on days 7 to 12 inclusive, of the metabolism trial. The samples were frozen immediately and dried later to a constant weight in a forced-draft oven at 70 C. They were then ground, and 10 g were taken fi'om each ground grab-sample and composited to provide one fecal sample per calf. The composite samples were stored for chemical analysis. Total urine from each calf was collected in 15 ml of 50% (vol/vol) H2SO4 for 48 h dur-
(21).
The main effects of each mineral, averaged over the combination of the other minerals, are presented with standard errors. Results and Discussion
It appeared (Table 3) that Zn had no effect on growth rate, Cu increased it by 5%, and Mn decreased it by 17%. No difference was significant (P < .05). Average daily gain was 1.42 kg. There were no significant differences (P < .05) in feed consumed per unit gain, and the values ranged from 3.3 kg to 4.1 kg feed per kg gain. Addition of Zn, Cu, or Mn had no effect (P > .05) on digestion coefficients of DM. Values ranged from 81.8% to 83.1% (Table 4). There were no appreciable differences be-
TABLE 2. Composition of the experimental rations. Ingredients
Control (1)
Zn (2)
Cu (3)
Basal ration (kg)
100
100
100
Z n S O , • 7rLO ( g ) CuSO, • 5~O (g)
...
MnSO4 • H20 (g) Chemical analysis Zn (ppm) Cu (ppm) Mn (ppm)
26.5
......
..
5.7
Zn q- Cu (4) 100 26.5 5.7
. . . . . . . . . . . . 40 6 12
100 6 12
40 20 12
101 21 12
Mn (5)
Zn + Mn Cu + Mn Zn + Cu + Zn (6) (7) (8)
100 ...
100 26.5
"ii.9
"ii.9
40 6 50
102 6 51
100 "" 5.7
1OO 26.5 5.7
40 21 49
97 19 47
11.9
11.9
JOURNAL OF DAIRY SCIENCE, VOL. 58, No. 3
412
IVAN AND GRIEVE
TABLE 3. Average daily gain and feed conversion. Gain per Feed per day kg gain
Treatment
- -
( 1) (2) (3) (4) (5 ) (6) (7) (8) Factor Zn
Control Zn Cu Zn -t- Cu Mn Zn q- Mn Cu -}- Mn Zn -t- Cu q- Mn
Cu Mn
Unsupplemented Supplemented Unsupplemented Supplemented Unsupplemented Supplemented
SE
(kg)
- -
1.44 1.62 1.59 1.55 1.29 1.16 1.34 1.31
3.5 3.9 4.1 3.9 3.4 3.9 3.3 3.6
1.42 1.41 1.38 1.45 1.55 1.28 .09
3.6 3.8 3.6 3.7 3.8 3.5 .1
tween treatments in the apparent digestibility of DM although the ration containing Zn -}Mn (treatment 6) had slightly lower coefficient than did the other rations. There were no differences between digestion coefficients that could be attributed to the addition of the trace minerals (Table 4). The apparent digestion coefficients of N and GE were lower in the ration with Zn and Mn than
in the other rations (Table 4), and the interaction between these minerals was significant (P < .05). There were no effects (P > .05) attributed to addition of trace minerals to the rations (Table 4). The highest percentage (47.0%) of NR was obtained for treatment 4 (basal qZn q- Cu) and the lowest (32.3%) for treatment 6 (basal + Zn + Mn). Since calves fed the ration supplemented with Zn or Mn had rates of gain, apparent digestion coefficients, and nitrogen retention comparable to those calves not fed these supplements, there was some interrelationship between Zn and Mn affecting feed utilization by calves. However, only interactions in digestibility of N and GE were significant. The lack of effect of supplemented Zn on digestibility of DM agrees with results by Miller et al. (18). Overall performance of calves and values for percentage NR in our experiment agree with those by Kehoe (10) using a similar high-concentrate ration. We expected that deficiencies of Zn and Cu would result in poorer feed efficiency (11, 4, 17, 19). Thompson (22) recommended 50 ppm, 7 ppm, and 30 ppm of Zn, Cu, and Mn in practical rations for cattle. The ARC (1) requirements suggest 50 ppm, 10 ppm, and 40 ppm of Zn, Cu, and Mn. Rations in this experiment contained 40 ppm, 6 ppm, and 12 ppm of Zn, Cu, and Mn before supplementa-
TABLE 4. Apparent digestibility of dry matter, nitrogen and gross energy, and nitrogen retention 0]9tained in various rations. Digestibility Treatment
Dry matter
Gross energy
Nitrogen retention
82.6 84.2 84.1 78.3 83.7 82.0
80.7 82.8 80.5 82.0 84.0 75.8 81.9 78.2
81.2 82.8 81.1 84.1 85.7 79.5 85.7 83.1
42.0 41.4 43.2
82.9 82.0 81.8 83.1 82.9 82.0 .9
81.8 79.7 80.8 80.7 81.5 80.0 1.1
83.4 82.4 82.3
42.4 38.5 40.2
83.5
40.6
82.3 83.5 .9
43.1 37.7 3.4
Nitrogen
(%) ( 1 ) Control (2) Zn (3) Cu (4) Zn -{- Cu (5) Mn (6) Zn q- Mn (7) Cu -}- Mn (8) Zu q- Cu -}- Mn Factor Unsupplemented Zn Supplemented Unsupplemented Cu Supplemented Unsupplemented Mn Supplemented SE JOURNAL OF DAIRY SCII~NCB, VOL. 58, NO. 3
81.5 83.4
47.0
46.4 32.3 38.1 34.3
413
T R A C E MINERALS F O R CALVES
TA~r,~. 5. Excretion of zinc, copper, and manganese in urine. Treatment (1) (2) (3) (4) (5) (6) (7) (8) Factor Zn
Zn
Control Zn Cu Zn + Cu Mn Zn+Mn Cu + Mn Zn + Cu + Mn
Unsupplemented Supplemented Cu Unsupplemented Supplemented Unstrpplemonted Mn Supplemented SE
Cu
($ of 2.59 .76 2.25 .45 .96 .85 1.63 1.47
Mn
daily intake) 10.67 .45 10.26 .39 16.49 .45 2.16 .33 1.41 .15 .95 .18 .84 .28 1.11 .22
1.86 .89* 1.29
1.45 1.52 1.23 .27
7.36 3.62** 5.83 5.15 9.90 1.08"* .43
.34 .29 .30 .33 .41 .21"* .03
*Differs from unsupplemented (P < .05). **Differs from unsupplemented (P < .01). tion with any of these minerals. These appeared adequate for satisfactory performanee of calves since addition of trace minerals did not improve it during a feeding period of 70 days. This agrees with Miller and Miller (16), who reported satisfactory weight gains when the ration contained 40 ppm Zn. Urinary excretion of Zn, as percent of intake
(Table 5), was reduced (P < .05) by the addition of Zn to the ration but was not affected (P > .05) by additional Cu or Mn, and none of the interactions was significant (P > .05). The reduction of urinary excret_ion of Zn due to the addition of Zn to the ration agrees with the report of Miller et aL (14). Furthermore, Miller et al. ( 15 )reported increased urinary excretion of ssZn in calves fed Zn-deficient ration. No deficiency of Zn was apparent in our experiment. Urinary excretion of Cu, as percent of intake, was reduced (P < .01) by supplemental Zn and Mn, but not by Cu (Table 5). However, Zn appeared to have this effect only when in combination with Cu (treatment 4 vs. treatment 2), Mn (treatment 6), or Cu and Mn (treatment 8). Manganese appeared to reduce Cu excretion when alone or in any combination with Zn or Cu. Urinary excretion of Mn, as percent of intake (Table 5), was low with all treatments. It was reduced (P < .01) by Mn supplementation of the diet but was not affected (P > .05) by Zn or Cu. None of the interactions was significant (P > .05). On the average, higher concentrations of Zn were in liver than in kidney and heart tissue (Table 6) on the dry matter basis. The addition of Zn and Cu to the rations did not affect concentrations of Zn in liver tissue although Cu appeared to result in a slight
TABLE 6. Concentration of zinc, copper, and mangan~e in liver, kidney, and heart (ppm DM). Manganese Treatment (1) (2) (3) (4) (5 ) (6) (7) (8)
Central Zn Cu Zn + Cu Mn Zn + Mn Cu + Mn Zn + Cu + Mn
Zinc concentration Liver Kidney Heart
Copper concentration Liver Kidney Heart
136 135 116 130 169 158 155 218
81 87 83 91 107 112 78 109
83 82 81 84 84 95 90 90
144 38 271 188 184 122 357 305
14.3 16.0 15.9 18.3 16.7 15.8 17.5 18.2
16.7 17.5 18.6 18.3 17.4 15.9 18.1 17.2
7.7 7.7 8.0 7.9 9.6 8.7 8.1 9.0
3.4 3.5 3.6 4.6 3.7 3.8 3.6 3.9
1.3 1.8 1.5 1.8 1.7 1.7 1.7 1.7
144 160 150 155 129 175" 11
87 100 97 90 85 102" 4
85 88 86 86 83 90** 1
239 163" 122 280** 160 242° 21
16.1 17.1 15.7 17.5 16.2 17.1 .7
17.7 17.3 16.9 18.1 17.8 17.2 .4
8.4 8.3 8.4 8.3 7.8 8.9* .3
3.6 4.0 36 3.9 3.8 3.8 .2
1.6 1.7 1.6 1.7 1.6 1.7 .1
concentration
Liver Kidney Heart
Factor Zn Cu Mn SE
Unsupplemented Supplemented Unsupplemented Supplemented Unsupplemented Supplemented
*Differs from unsuppl~aented (P < .05). **Differs from unsupplemented (P < .01). JOURNAL OF DAIRY SCIENCR, VOL. 78, NO. 3
414
~ANA~DGmEVE
decrease. The addition of Mn, and particular- tration of Cu in livers of calves fed low Cu and ly in combination with Zn and Cu, appeared high Zn in the ration (treatment 2) indicated to increase the concentration of Zn in liver tis- that deficiency could have developed ff the exsue. periment had continued for a longer period. Supplemental Zn or Cu did not affect Therefore, it appears that high dietary Zn the concentration of Zn in kidney tissue (Ta- m/ght be useful in prevention of copper toxicible 6). However, there was an increase ty in cattle. (P < .05) in Zn concentration associated with There were no appreciable effects on Gu Mn supplementation. Manganese supplementa- concentrations in kidney or heart associated tion also increased (P < .01) Zn concentra- with the addition of the trace minerals to the tion in heart tissue. rations. The concentrations of Mn in liver, kidney, Miller eL al. (14) reported a significant increase in the Zn concentration in liver when and heart tissue (Table 6) were lower than a ration containing 33 ppm Zn was increased those of Zn or Cu. The addition of Zn and Cu in Zn content to 233 ppm, but no effect on Zn to the ration had little effect on tissue concenconcentrations in kidney and heart tissues. The trations of Mn, but the addition of Mn increased (P < .05) the liver tissue concentration of lack of increase in the Zn concentration in the liver in this experiment could be caused by the Mn. Therefore, higher dietary Mn increased its relatively low, (two-fold) increase in Zn in the net retention though kidneff and hearL tissue ration as compared with the seven-fold in- concentrations were not affected. Howes and Dyer (9) reported a similar action of dietary crease used by Miller et al. (14). The concentration of C u in liver tissue was Mn in calves at 7 days of age. In this experiment, it appeared that a highmuch higher than in kidney or heart tissue (Table 6). The addition of Zn to the ration re- concentrate ration could be fed to fattening suited in a marked reduction in the concentra- bull-calves without the trace mineral suppletion of Cu whereas Mn resulted in some in- ments. The formulation of the ration should crease, and Cu resulted in a marked increase be balanced for all trace minerals; Cu should in the liver concentration of Cu. When Zn was be increased ff there is a high content of Zn in added with Cu, there was increase in the liver the ration, and both Zn and Cu should be kept concentration of Cu, but the highest was when low ff there is a high content of Mn in the raCu and Mn were added together. The addi- tion. tion of Zn, Cu, and Mn also resulted in a high Acknowledgments liver concentration of Cu. Zn was associated We thank R. T. Hardin for assistance with with a decrease (P < .05), Cu was associated with an increase (P < .01), and Mrt was as- statistical analysis and S. Nielsen for care of sociated with an increase (P < .05) in the experimental animals. A research assistantship provided by the Naliver concentration of Cu. It appeared that Zn in the diet decreased the liver concentration tional Research Council is gratehzlly acknowlof Cu, that Mn partially offset the depressing edged. effect of Zn and that Mn increased the reReferences sponse to supplemental Cu. (1) Agricultural Research Council. 1965. The The inhibitory effect of higher dietary Zn nutrient requirements of farm livestock. upon the Cu concentration in the liver tissue No. 2. Ruminants. Agrieulhtral Research was also found by other workers, (20, 13, 5). Council, London. The significant increase in liver concentration (~) Association o~ Official Agricultural Chemof Cu, associated with its higher dietary ists. 1965. Official methods of analysis. 10th amounts, agrees with reports of Dick (6) and Ed. Washington, IX:. Gartner et al. (7). Since higher dietary Mn (3) Bentley, O. C., and. P. H. Phillips. 1951. was associated with increase in liver concentraThe effect of low manganese rations upon dairy cattle. J. Dairy SCI. 34:396. tion and a ten-fold decrease in urinary excre(4) Blackrnon, D. M., W. J. Miller, and J. D. tion of Cu, this suggests that higher dietary Morton. 1967. Zinc deficiency in ruminants, Mr[ increased the incidence of binding sites of occurrence, effects, diagnosis, treatments. Cu in the liver, thereby increasing incorporaVet. Med. 62:265. tion of Cu into the liver and decreasing its (5) Davis, G. K. 1958. M~laauism of trace eleelimination in the urine. ment hmetion. Soil. Sci. 85:59. There was no evidence of Cu deficiency, but (6) Dick, A. T. 1954. Studies on the assimilahigher supplemental Zn in a ration containing tion and storage of copper in crossbred low Cu could create it. The very low concertsheep. Australian J. Agr. Res. 5:511. JOURNAL OF DAIRY SCIENCE, "V'OL. 58. NO. 5
TRACE MINERALS FOR CALVES
(7) Gartner, It, J. W., J. G. Young, and P. M. Pepper. 1968. Hepatic copper concentration of steers grazing pastures on developed wet heath land in south-eastern Queensland. Australiau J. Exp. Agr. Anita. Husb. 8:679. (8) Hill, F. W., and D. L. Anderson. 1958. Comparison of metabolizable energy and productive energy determinations with growing chicks. J. Nutr. 64:587. (9) Howes, A. D., and I. A. Dyer. 1971. Diet and supplemental mineral effects on manganese metabolism in newborn calves. J. Anita. Sei. 32:141. (10) Kehoo, F. X. 1969. Master of Science Thesis. University o.f Alberta, Edmonton, Alberta. (11) Lamand, M., 1. Bel]anger, and Y. Geay. 1969. Copper deficiency in fattening bull calves. Influence on growth and treatment. Ann. Zootech. (Paris) 18:227. (12) Maynard, L. A., and J. K. Loosli. 1962. Animal nutrition, 5th Ed. McGraw-H2ll Book Co., Inc., New York. (13) McCall, J. T., and G. K. Davis. 1961.. Effect of dietary protein and zinc on the absorption and liver deposition of radioactive and total copper. I- Nutr. 74:45. (14) Miller, W. 1., D. M. Blaekrnon, R. P. Gentry, and F. M. Pate. 1970. Effects of high but nontoxic levels of zinc in practical diets on ~Zn and zinc metabolism in Holstein calves. ]. Nutr. 100:893.
415
15) Miller, W. l., D. M. Blackmon, G. W. Pewell, R. P. Gentry, and J. M. Hiers, Jr. 1966. Effects of zinc deficiency per se and of dietary zinc level on urinary and endoge~aous fecal excretion of ~ n fi-om a single intravenous dose by ruminants. J. Nutr. 90:335. 16) Miller, J. K., and W. J. Miller. 1962. Experimental zinc deficiency and recovery of calves. 1. Nutr. 76:467. 17) Miller, W. J., W. 1. Pitts, C. M. Clifton, and J. D. Morton. 1965. Effects of zinc deficiency per se on feed efficiency, Serum Alkaline Phosphatase zinc in skin, behaviour, greying, and other measuremer~ts in the Holstein calf. 1. Dairy Sei. 48:1329. (18) Miller, W. 1., G. W. Powell, and I- M. Heirs, Jr. 1966. Influence of zinc deficiency on dry matter digestibility in ruminants. J. Dairy Sei. 49:1012. (19) Neal, W. M., R. B. Becker, and A. L. Shealy. 1931. A natural copper deficiency in cattle rations. Science 74:418. (20) Ritehie, H. D., R. W. Luecke, B. V. Baltzer, E. R. Miller, D. E. Ullrey, and J. A. Heeler. 1963. Copper and zinc interrelationships in the pig. J. Nutr. 79:117. (21) Smillie, K. W. 1969. Statpak 2: an APL statistical package, 2nd Ed. Department of Computing Science, University of Alberta, Edmonton, Alberta. 17:34. (22) Thompson, D. J. 1970. Trace elements in animal nutrition. International Minerals & Chemical Corporation, Skokie, IL 60076.
JOURNAL OF DAIRY SCIENCE, VOL. 58, No. 3
Deficiencies of Zn (4, 17), Cu (11, 19), and Mn (3) decreased growth rate, which was improved by addition of the deficient mineral to the diet. Studies of effects of these individual trace minerals on performance of animals axe of little value because of interactions between the minerals. The main objective of our experiment was to study effects of addition of three trace minerals (Zn, Cu, and Mn) to a practical highconcentrate ration on the performance of bull calves. A second objective was to study interactions among these trace minerals. For all diets, measurements were of growth rate, efficiency of feed utilization, digestibility of dry matter (DM), nitrogen (N) and gross energy (GE), nitrogen retention (NR), excretion of Zn, Cu, and Mn in urine, and concentrations of minerals in liver, heart, and kidney tissues.
The effects of supplemental zinc, copper, and manganese alone or in combination in a high-concentrate ration were studied in 16 Holstein bull calves during a 10-wk feeding trial. Metabolism was studied after the feeding trial. Apparent digestibility of dry matter, nitrogen, and gross energy, and nitrogen retention and urinary excretion of zinc, copper, and manganese were determined. The calves were slaughtered after the experiment, and liver, heart, mad kidney were taken for analyses of trace minerals. Supplementation o~ the basal ration with the trace minerals did not affect body weight gains, which averaged 1.42 kg daily. The addition of trace minerals did not affect apparent digestibility coefficients. A zinc-manganese interaction in digestion of nitrogen and gross energy was significant. Higher dietary manganese caused increased zinc concentrations in the liver, kidney, and heart. The copper coneentratien of liver was decreased by dietary zinc and increased by dietary copper and manganese. Supplemental manganese increased its net retention. There was no evidence of deficiency of any trace minerals in the unsupplemented treatments.
Materials and Methods
Introduction Trace minerals are usually added to cattle rations to prevent or overcome deficiencies and
possible consequences on animal performance. The need for trace mineral supplementation has not been defined because of wide variation between feeds in their mineral content. This variation is due, in part at least, to the amount of elements in the soil, availability o~ soil minerals to the plants grown, and availability of plant minerals to animals fed. ,Received September 3, 1974. a Presemt address: Animal Research Institute,
Agriculture Canada, Ottawa, Ontario, Canada K1A 0(26.
Sixteen Holstein bull calves were allotted by weight in a 2 X 2 X 2 factorial design to eight treatments; (1) basal ration, (2) basal + Zn, (3) basal + Cu, (4) basal q- Zn _i_ Cu, (5) basal + Mn, (6) basal -t- Zn -t- Mn, (7) basal q- Cu + Mn, and (8) basal -b Zn q- (3u -k Mn. Calves in each treatment varied in average initial age from 22 to 28 wk and in average initial liveweight from 111 to 168 kg. They were in individual stalls and fed a d libitum. Water and a mixture of equal weights of calcium phosphate and eobaltized-iodized salt were available free choice. Ini6al and final individual liveweights were from two successive weighings at 24-h intervals after the calves were without feed and water overnight. The basal ration (Table 1) was formulated for all eight treatments and consisted primarily of coarsely ground barley. Urea was ineluded to provide a calculated 12% of protein equivalent. Experimental rations (Table 2) were formulated by addition of salts of particular trace minerals to the basal ration. Trace mineral salts were added to provide I00 ppm Zn, 20 ppm Cu, and 50 ppm Mn in the ration. Barley con-
410
411
TRACE MIN'E,RALS FOR CALVES
ing the period of metabolism study. A 5% aliquot was collected twice daily in a polyethylene bottle and stored at 4 G for chemical analysis. The calves were slaughtered the day after metabolism studies were completed. The heart, right kidney, and samples o f liver from each calf were taken. These samples were dried in a freeze-drier for 3 days, then homogenized in a laboratory blender and stored for analysis of trace minerals. The methods of AOAC (2) were used for determination of DM in the feed and fecal samples and of N in the feed, fecal, and urine samples. Gross energy in feed and fecal samples was measured by combustion in a Parr oxygen bomb calorimeter. Determination of Cr~Os in the feed and fecal samples was according to the method of Hill and Anderson (8). Atomic Absorption Spectrophotometry (Techtror~ AA-3) was used for analyses of Cu, Zn, and Mn. Samples of feed, liver, kidney, heart, and freeze-dried urine were prepared by wet ashing as outlined in AOAC (2). Statistical analyses were on the IBM 360/ 67 computer by a program written by Smillie
TABLE 1. Composition of the basal ration. ~e~e~s
%
Barley Urea Limestone Cobaltized-Iodized salt Vitamin A (10,000 IU/g) Vitamin D (35,000 IU/g) Vitamin E (44 IU/g) Chemical analysis Dry matter (%) Crude protein (~) Gross energy (Meal/kg) Calcium (%) Phosphorus (%)
97.6 .8 1.0 .5 .088 .0055 .015 88.1 12.6 4.12 .46 .40
tained 40 ppm, 6 ppm, and 12 ppm of Zn, Cu, and Mn. Metabolism was studied after the 10-wk feeding was completed. Chromic oxide (Cr2Os) was the indicator of fecal excretion (12). Each ration fed to experimental animals had .5% of Cr2Os mixed into it. To ensure complete consumption of the daily ration, feed offered was restricted by .5 kg below the average daily consumption for each calf during the final week of the feeding period. Daily rations were divided into two parts and fed twice daily for 12 days. A 5-g sample of feed was taken at each feeding. This was composited to form one sample per each ration, ground, and stored for analysis. Fecal grab-samples were obtained four times daily, on days 7 to 12 inclusive, of the metabolism trial. The samples were frozen immediately and dried later to a constant weight in a forced-draft oven at 70 C. They were then ground, and 10 g were taken fi'om each ground grab-sample and composited to provide one fecal sample per calf. The composite samples were stored for chemical analysis. Total urine from each calf was collected in 15 ml of 50% (vol/vol) H2SO4 for 48 h dur-
(21).
The main effects of each mineral, averaged over the combination of the other minerals, are presented with standard errors. Results and Discussion
It appeared (Table 3) that Zn had no effect on growth rate, Cu increased it by 5%, and Mn decreased it by 17%. No difference was significant (P < .05). Average daily gain was 1.42 kg. There were no significant differences (P < .05) in feed consumed per unit gain, and the values ranged from 3.3 kg to 4.1 kg feed per kg gain. Addition of Zn, Cu, or Mn had no effect (P > .05) on digestion coefficients of DM. Values ranged from 81.8% to 83.1% (Table 4). There were no appreciable differences be-
TABLE 2. Composition of the experimental rations. Ingredients
Control (1)
Zn (2)
Cu (3)
Basal ration (kg)
100
100
100
Z n S O , • 7rLO ( g ) CuSO, • 5~O (g)
...
MnSO4 • H20 (g) Chemical analysis Zn (ppm) Cu (ppm) Mn (ppm)
26.5
......
..
5.7
Zn q- Cu (4) 100 26.5 5.7
. . . . . . . . . . . . 40 6 12
100 6 12
40 20 12
101 21 12
Mn (5)
Zn + Mn Cu + Mn Zn + Cu + Zn (6) (7) (8)
100 ...
100 26.5
"ii.9
"ii.9
40 6 50
102 6 51
100 "" 5.7
1OO 26.5 5.7
40 21 49
97 19 47
11.9
11.9
JOURNAL OF DAIRY SCIENCE, VOL. 58, No. 3
412
IVAN AND GRIEVE
TABLE 3. Average daily gain and feed conversion. Gain per Feed per day kg gain
Treatment
- -
( 1) (2) (3) (4) (5 ) (6) (7) (8) Factor Zn
Control Zn Cu Zn -t- Cu Mn Zn q- Mn Cu -}- Mn Zn -t- Cu q- Mn
Cu Mn
Unsupplemented Supplemented Unsupplemented Supplemented Unsupplemented Supplemented
SE
(kg)
- -
1.44 1.62 1.59 1.55 1.29 1.16 1.34 1.31
3.5 3.9 4.1 3.9 3.4 3.9 3.3 3.6
1.42 1.41 1.38 1.45 1.55 1.28 .09
3.6 3.8 3.6 3.7 3.8 3.5 .1
tween treatments in the apparent digestibility of DM although the ration containing Zn -}Mn (treatment 6) had slightly lower coefficient than did the other rations. There were no differences between digestion coefficients that could be attributed to the addition of the trace minerals (Table 4). The apparent digestion coefficients of N and GE were lower in the ration with Zn and Mn than
in the other rations (Table 4), and the interaction between these minerals was significant (P < .05). There were no effects (P > .05) attributed to addition of trace minerals to the rations (Table 4). The highest percentage (47.0%) of NR was obtained for treatment 4 (basal qZn q- Cu) and the lowest (32.3%) for treatment 6 (basal + Zn + Mn). Since calves fed the ration supplemented with Zn or Mn had rates of gain, apparent digestion coefficients, and nitrogen retention comparable to those calves not fed these supplements, there was some interrelationship between Zn and Mn affecting feed utilization by calves. However, only interactions in digestibility of N and GE were significant. The lack of effect of supplemented Zn on digestibility of DM agrees with results by Miller et al. (18). Overall performance of calves and values for percentage NR in our experiment agree with those by Kehoe (10) using a similar high-concentrate ration. We expected that deficiencies of Zn and Cu would result in poorer feed efficiency (11, 4, 17, 19). Thompson (22) recommended 50 ppm, 7 ppm, and 30 ppm of Zn, Cu, and Mn in practical rations for cattle. The ARC (1) requirements suggest 50 ppm, 10 ppm, and 40 ppm of Zn, Cu, and Mn. Rations in this experiment contained 40 ppm, 6 ppm, and 12 ppm of Zn, Cu, and Mn before supplementa-
TABLE 4. Apparent digestibility of dry matter, nitrogen and gross energy, and nitrogen retention 0]9tained in various rations. Digestibility Treatment
Dry matter
Gross energy
Nitrogen retention
82.6 84.2 84.1 78.3 83.7 82.0
80.7 82.8 80.5 82.0 84.0 75.8 81.9 78.2
81.2 82.8 81.1 84.1 85.7 79.5 85.7 83.1
42.0 41.4 43.2
82.9 82.0 81.8 83.1 82.9 82.0 .9
81.8 79.7 80.8 80.7 81.5 80.0 1.1
83.4 82.4 82.3
42.4 38.5 40.2
83.5
40.6
82.3 83.5 .9
43.1 37.7 3.4
Nitrogen
(%) ( 1 ) Control (2) Zn (3) Cu (4) Zn -{- Cu (5) Mn (6) Zn q- Mn (7) Cu -}- Mn (8) Zu q- Cu -}- Mn Factor Unsupplemented Zn Supplemented Unsupplemented Cu Supplemented Unsupplemented Mn Supplemented SE JOURNAL OF DAIRY SCII~NCB, VOL. 58, NO. 3
81.5 83.4
47.0
46.4 32.3 38.1 34.3
413
T R A C E MINERALS F O R CALVES
TA~r,~. 5. Excretion of zinc, copper, and manganese in urine. Treatment (1) (2) (3) (4) (5) (6) (7) (8) Factor Zn
Zn
Control Zn Cu Zn + Cu Mn Zn+Mn Cu + Mn Zn + Cu + Mn
Unsupplemented Supplemented Cu Unsupplemented Supplemented Unstrpplemonted Mn Supplemented SE
Cu
($ of 2.59 .76 2.25 .45 .96 .85 1.63 1.47
Mn
daily intake) 10.67 .45 10.26 .39 16.49 .45 2.16 .33 1.41 .15 .95 .18 .84 .28 1.11 .22
1.86 .89* 1.29
1.45 1.52 1.23 .27
7.36 3.62** 5.83 5.15 9.90 1.08"* .43
.34 .29 .30 .33 .41 .21"* .03
*Differs from unsupplemented (P < .05). **Differs from unsupplemented (P < .01). tion with any of these minerals. These appeared adequate for satisfactory performanee of calves since addition of trace minerals did not improve it during a feeding period of 70 days. This agrees with Miller and Miller (16), who reported satisfactory weight gains when the ration contained 40 ppm Zn. Urinary excretion of Zn, as percent of intake
(Table 5), was reduced (P < .05) by the addition of Zn to the ration but was not affected (P > .05) by additional Cu or Mn, and none of the interactions was significant (P > .05). The reduction of urinary excret_ion of Zn due to the addition of Zn to the ration agrees with the report of Miller et aL (14). Furthermore, Miller et al. ( 15 )reported increased urinary excretion of ssZn in calves fed Zn-deficient ration. No deficiency of Zn was apparent in our experiment. Urinary excretion of Cu, as percent of intake, was reduced (P < .01) by supplemental Zn and Mn, but not by Cu (Table 5). However, Zn appeared to have this effect only when in combination with Cu (treatment 4 vs. treatment 2), Mn (treatment 6), or Cu and Mn (treatment 8). Manganese appeared to reduce Cu excretion when alone or in any combination with Zn or Cu. Urinary excretion of Mn, as percent of intake (Table 5), was low with all treatments. It was reduced (P < .01) by Mn supplementation of the diet but was not affected (P > .05) by Zn or Cu. None of the interactions was significant (P > .05). On the average, higher concentrations of Zn were in liver than in kidney and heart tissue (Table 6) on the dry matter basis. The addition of Zn and Cu to the rations did not affect concentrations of Zn in liver tissue although Cu appeared to result in a slight
TABLE 6. Concentration of zinc, copper, and mangan~e in liver, kidney, and heart (ppm DM). Manganese Treatment (1) (2) (3) (4) (5 ) (6) (7) (8)
Central Zn Cu Zn + Cu Mn Zn + Mn Cu + Mn Zn + Cu + Mn
Zinc concentration Liver Kidney Heart
Copper concentration Liver Kidney Heart
136 135 116 130 169 158 155 218
81 87 83 91 107 112 78 109
83 82 81 84 84 95 90 90
144 38 271 188 184 122 357 305
14.3 16.0 15.9 18.3 16.7 15.8 17.5 18.2
16.7 17.5 18.6 18.3 17.4 15.9 18.1 17.2
7.7 7.7 8.0 7.9 9.6 8.7 8.1 9.0
3.4 3.5 3.6 4.6 3.7 3.8 3.6 3.9
1.3 1.8 1.5 1.8 1.7 1.7 1.7 1.7
144 160 150 155 129 175" 11
87 100 97 90 85 102" 4
85 88 86 86 83 90** 1
239 163" 122 280** 160 242° 21
16.1 17.1 15.7 17.5 16.2 17.1 .7
17.7 17.3 16.9 18.1 17.8 17.2 .4
8.4 8.3 8.4 8.3 7.8 8.9* .3
3.6 4.0 36 3.9 3.8 3.8 .2
1.6 1.7 1.6 1.7 1.6 1.7 .1
concentration
Liver Kidney Heart
Factor Zn Cu Mn SE
Unsupplemented Supplemented Unsupplemented Supplemented Unsupplemented Supplemented
*Differs from unsuppl~aented (P < .05). **Differs from unsupplemented (P < .01). JOURNAL OF DAIRY SCIENCR, VOL. 78, NO. 3
414
~ANA~DGmEVE
decrease. The addition of Mn, and particular- tration of Cu in livers of calves fed low Cu and ly in combination with Zn and Cu, appeared high Zn in the ration (treatment 2) indicated to increase the concentration of Zn in liver tis- that deficiency could have developed ff the exsue. periment had continued for a longer period. Supplemental Zn or Cu did not affect Therefore, it appears that high dietary Zn the concentration of Zn in kidney tissue (Ta- m/ght be useful in prevention of copper toxicible 6). However, there was an increase ty in cattle. (P < .05) in Zn concentration associated with There were no appreciable effects on Gu Mn supplementation. Manganese supplementa- concentrations in kidney or heart associated tion also increased (P < .01) Zn concentra- with the addition of the trace minerals to the tion in heart tissue. rations. The concentrations of Mn in liver, kidney, Miller eL al. (14) reported a significant increase in the Zn concentration in liver when and heart tissue (Table 6) were lower than a ration containing 33 ppm Zn was increased those of Zn or Cu. The addition of Zn and Cu in Zn content to 233 ppm, but no effect on Zn to the ration had little effect on tissue concenconcentrations in kidney and heart tissues. The trations of Mn, but the addition of Mn increased (P < .05) the liver tissue concentration of lack of increase in the Zn concentration in the liver in this experiment could be caused by the Mn. Therefore, higher dietary Mn increased its relatively low, (two-fold) increase in Zn in the net retention though kidneff and hearL tissue ration as compared with the seven-fold in- concentrations were not affected. Howes and Dyer (9) reported a similar action of dietary crease used by Miller et al. (14). The concentration of C u in liver tissue was Mn in calves at 7 days of age. In this experiment, it appeared that a highmuch higher than in kidney or heart tissue (Table 6). The addition of Zn to the ration re- concentrate ration could be fed to fattening suited in a marked reduction in the concentra- bull-calves without the trace mineral suppletion of Cu whereas Mn resulted in some in- ments. The formulation of the ration should crease, and Cu resulted in a marked increase be balanced for all trace minerals; Cu should in the liver concentration of Cu. When Zn was be increased ff there is a high content of Zn in added with Cu, there was increase in the liver the ration, and both Zn and Cu should be kept concentration of Cu, but the highest was when low ff there is a high content of Mn in the raCu and Mn were added together. The addi- tion. tion of Zn, Cu, and Mn also resulted in a high Acknowledgments liver concentration of Cu. Zn was associated We thank R. T. Hardin for assistance with with a decrease (P < .05), Cu was associated with an increase (P < .01), and Mrt was as- statistical analysis and S. Nielsen for care of sociated with an increase (P < .05) in the experimental animals. A research assistantship provided by the Naliver concentration of Cu. It appeared that Zn in the diet decreased the liver concentration tional Research Council is gratehzlly acknowlof Cu, that Mn partially offset the depressing edged. effect of Zn and that Mn increased the reReferences sponse to supplemental Cu. (1) Agricultural Research Council. 1965. The The inhibitory effect of higher dietary Zn nutrient requirements of farm livestock. upon the Cu concentration in the liver tissue No. 2. Ruminants. Agrieulhtral Research was also found by other workers, (20, 13, 5). Council, London. The significant increase in liver concentration (~) Association o~ Official Agricultural Chemof Cu, associated with its higher dietary ists. 1965. Official methods of analysis. 10th amounts, agrees with reports of Dick (6) and Ed. Washington, IX:. Gartner et al. (7). Since higher dietary Mn (3) Bentley, O. C., and. P. H. Phillips. 1951. was associated with increase in liver concentraThe effect of low manganese rations upon dairy cattle. J. Dairy SCI. 34:396. tion and a ten-fold decrease in urinary excre(4) Blackrnon, D. M., W. J. Miller, and J. D. tion of Cu, this suggests that higher dietary Morton. 1967. Zinc deficiency in ruminants, Mr[ increased the incidence of binding sites of occurrence, effects, diagnosis, treatments. Cu in the liver, thereby increasing incorporaVet. Med. 62:265. tion of Cu into the liver and decreasing its (5) Davis, G. K. 1958. M~laauism of trace eleelimination in the urine. ment hmetion. Soil. Sci. 85:59. There was no evidence of Cu deficiency, but (6) Dick, A. T. 1954. Studies on the assimilahigher supplemental Zn in a ration containing tion and storage of copper in crossbred low Cu could create it. The very low concertsheep. Australian J. Agr. Res. 5:511. JOURNAL OF DAIRY SCIENCE, "V'OL. 58. NO. 5
TRACE MINERALS FOR CALVES
(7) Gartner, It, J. W., J. G. Young, and P. M. Pepper. 1968. Hepatic copper concentration of steers grazing pastures on developed wet heath land in south-eastern Queensland. Australiau J. Exp. Agr. Anita. Husb. 8:679. (8) Hill, F. W., and D. L. Anderson. 1958. Comparison of metabolizable energy and productive energy determinations with growing chicks. J. Nutr. 64:587. (9) Howes, A. D., and I. A. Dyer. 1971. Diet and supplemental mineral effects on manganese metabolism in newborn calves. J. Anita. Sei. 32:141. (10) Kehoo, F. X. 1969. Master of Science Thesis. University o.f Alberta, Edmonton, Alberta. (11) Lamand, M., 1. Bel]anger, and Y. Geay. 1969. Copper deficiency in fattening bull calves. Influence on growth and treatment. Ann. Zootech. (Paris) 18:227. (12) Maynard, L. A., and J. K. Loosli. 1962. Animal nutrition, 5th Ed. McGraw-H2ll Book Co., Inc., New York. (13) McCall, J. T., and G. K. Davis. 1961.. Effect of dietary protein and zinc on the absorption and liver deposition of radioactive and total copper. I- Nutr. 74:45. (14) Miller, W. 1., D. M. Blaekrnon, R. P. Gentry, and F. M. Pate. 1970. Effects of high but nontoxic levels of zinc in practical diets on ~Zn and zinc metabolism in Holstein calves. ]. Nutr. 100:893.
415
15) Miller, W. l., D. M. Blackmon, G. W. Pewell, R. P. Gentry, and J. M. Hiers, Jr. 1966. Effects of zinc deficiency per se and of dietary zinc level on urinary and endoge~aous fecal excretion of ~ n fi-om a single intravenous dose by ruminants. J. Nutr. 90:335. 16) Miller, J. K., and W. J. Miller. 1962. Experimental zinc deficiency and recovery of calves. 1. Nutr. 76:467. 17) Miller, W. J., W. 1. Pitts, C. M. Clifton, and J. D. Morton. 1965. Effects of zinc deficiency per se on feed efficiency, Serum Alkaline Phosphatase zinc in skin, behaviour, greying, and other measuremer~ts in the Holstein calf. 1. Dairy Sei. 48:1329. (18) Miller, W. 1., G. W. Powell, and I- M. Heirs, Jr. 1966. Influence of zinc deficiency on dry matter digestibility in ruminants. J. Dairy Sei. 49:1012. (19) Neal, W. M., R. B. Becker, and A. L. Shealy. 1931. A natural copper deficiency in cattle rations. Science 74:418. (20) Ritehie, H. D., R. W. Luecke, B. V. Baltzer, E. R. Miller, D. E. Ullrey, and J. A. Heeler. 1963. Copper and zinc interrelationships in the pig. J. Nutr. 79:117. (21) Smillie, K. W. 1969. Statpak 2: an APL statistical package, 2nd Ed. Department of Computing Science, University of Alberta, Edmonton, Alberta. 17:34. (22) Thompson, D. J. 1970. Trace elements in animal nutrition. International Minerals & Chemical Corporation, Skokie, IL 60076.
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