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Interrelationship of Dietary Vitamin D3 with Potassium, Sodium and Magnesium in Young Turkeys1,2
482
J. B. COOPER
REFERENCES
lowest for treatment 4, Table 1. Treatment 4 also resulted in the lowest number of squabs being produced per pair for any of the five treatments. Treatment 5 (open lofts)
was
second lowest. ACKNOWLEDGEMENTS The author is indebted to Dr. Walter E.
Levi, W. M., 1941. The Pigeon. R. L. Bryan Co., Columbia, S.C. Platt, C. S., and R. S. Dare, 1937. New Jersey State Pigeon Breeding Test. New Jersey Agri. Exp. Sta. Bui. 634. Steel, R. G. D„ and J. H. Torrie, 1960. Principles and Procedures of Statistics. McGraw-Hill, New York.
Johnston of the Experimental Statistical Services of Clemson University for the statistical analysis.
AYHAN A K S O Y 3 AND T . W . SULLIVAN
Department of Poultry & Wildlife Sciences, University of Nebraska, Lincoln, Nebraska 68583 (Received for publication June 28, 1976)
ABSTRACT Six experiments were conducted with Large White and Bronze turkeys to four weeks of age. The interrelationship of dietary vitamin D 3 with either potassium, sodium or magnesium was studied in two experiments each. One group of 8-9 male and one group of 8-9 female poults were randomly assigned to each treatment within an experiment. The design of all experiments involved a factorial arrangement of two variables: vitamin D 3 and a mineral element. For example, all possible combinations of four dietary potassium levels, 0.24, 0.34, 0.44, 0.54%; and four vitamin D 3 levels 400, 800, 1600 and 3200 I.C. units/kg. were fed in the second potassium experiment. Significant (P < 0.05) weight gain differences occurred among both K levels and D 3 levels in this particular experiment. Average four-week body weights were 336, 393, 382 and 430 g., respectively, for potassium levels and 330, 391, 404 and 426 g., respectively, for vitamin D 3 levels. There was a significant interaction between dietary K and D 3 levels relative to four-week body weight gains. Vitamin D 3 tended to increase retention of both potassium and sodium in the body. A significant interaction occurred between dietary magnesium and vitamin D 3 relative to body weight gain. Vitamin D 3 increased magnesium content in tibiae ash and decreased magnesium content in serum. POULTRY SCIENCE 56: 482-491, 1977
INTRODUCTION
T
HE effect of vitamin D on calcium metabolism has been extensively studied but
little is known about its effect on the metabo-
lism of other cations. A few studies have indicated that vitamin D plays a role in the utilization of other cations. As early as 1930 McHargue et al. reported that the carcass magnesium content of rats subjected to ultraviolet irradiation was less than that of control
1. Published as paper Number 5142, Journal Series, Nebraska Agricultural Experiment Station. 2. From a thesis submitted by the senior author in partial fulfillment of the requirements for the Ph.D. degree. 3. Present address: Animal Science Department, College of Agriculture, Ataturk University, Erzurum, Turkey.
animals.
Meintzer
and
Steenbock
(1956)
showed that vitamin D supplementation increased magnesium absorption in rats. Similar results were obtained by Wasserman (1962) with chicks. Worker and Migicovsky (1961) reported that vitamin D 3 significantly creased the deposition of
28
in-
Mg in the tibiae
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Interrelationship of Dietary Vitamin D 3 with Potassium, Sodium and Magnesium in Young Turkeys1-2
483
VITAMIN D 3 X K, NA AND MG IN POULT DIETS
EXPERIMENTAL PROCEDURE Six experiments were conducted with Large White and Bronze turkeys, day-old to four weeks of age. A factorial arrangement of dietary treatments, involving all combinations of three or four vitamin D 3 levels and three or four levels of the particular mineral element, was used in each experiment. All poults were maintained in thermostatically controlled, electrically heated battery brooders with raised wire floors throughout the experimental period. The experimental diet and water were supplied ad libitum. Determination of sodium in basal diet, feces and blood serum; potassium in basal diet and feces; magnesium in basal diet, blood serum and tibiae were made by methods described by Hassan and Knudsen (1968). Calcium and phosphorus levels in all basal diets were maintained somewhat higher than known requirements. It was hoped that moderate excesses of dietary Ca and P would minimize the effect of low D 3 levels on the absorption and metabolism of these elements and overall poult performance. All diets were
supplemented with penicillin-streptomycin (1:3) to provide 22 parts per million (p.p.m.) of the antibiotics. Four-week body weight data were subjected to analysis of variance (Steel and Torrie, 1960) and Duncan's (1955) multiple range test. Individual bird data were used in these statistical procedures. Feed efficiency and survival data were not analyzed statistically because true replicates were not involved in TABLE 1.—Composition of the basal diet for potassium and magnesium experiments' Ingredients
Per kg. of diet
Dextrose sugar (cerelose) Isolated soybean protein Solka Floe (wood flour) Corn oil Methionine-hydroxy-analog Glycine Ca3(P04)2 NaCl MgS04-7H202 FeC6H507-5H20 ZnS04-7H20 CuS04-5H20 KC1 3 MnS04H20 KI Choline chloride (50%) Myvamix (44 I.U. vit. E/g.) Vitamin A premix (5,000 I.U./g.) Vitamin B premix in sucrose 4 Penicillin-streptomycin 5
463.00 370.00 40.00 45.00 5.00 2.00 40.00 5.00 4.80 0.60 0.44 0.04 8.00 0.38 0.04 5.00 1.00 3.00 5.00 0.50
g-
'Calculated average composition: Protein, % 30.0 Metabolizable energy, Kcal./kg. 3046 Calcium, % 1.57 Phosphorus, % 1.05 2 Omitted from the basal diet in magnesium experiments. 3 Omitted from the basal diet in potassium experiments. "Vitamin B premix provided the following per kg. of diet: 0.02 mg. vitamin B12,0.30 mg. d-biotin, 4 mg. menadione, 8 mg. pyridoxine, 6 mg. folic acid, 10 mg. riboflavin, 30 mg. calcium pantothenate, 15 mg. thiamine HC1, and 100 mg. niacin. 5 Pro-Strep (Merck and Company) contains 44 mg. of penicillin-streptomycin, 1:3 per g.
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of chickens. Vitamin D treatment caused hypomagnesemia in rats (Hanna 1961; Richardson and Welt, 1965; Lifshitz et al., 1967). Vitamin D apparently affects the absorption and metabolism of potassium and sodium to a lesser extent than that of magnesium. Wasserman (1962) found that vitamin D 3 had no significant influence on absorption or tissue distribution of sodium and potassium in chicks with ligated duodenums. Large doses of vitamin D 2 given intramuscularly in rats had no influence on the sodium level of blood. But a lower plasma level of potassium was obtained following the administration of calciferol. The present investigation was undertaken to study possible interrelationships between dietary vitamin D 3 , sodium, potassium and magnesium in young turkeys.
484
A. AKSOY AND T. W. SULLIVAN
the experiments. Tissue and fecal composition data were evaluated by analysis of variance and Duncan's multiple range test.
TABLE 2.—Composition of basal diet for sodium experiments' Ingredients
Experiment 2. The basal diet, which was the same as used in experiment 1, was supplemented with 600, 1200, 1800 and 3600 I.C. units of vitamin D 3 per kg., and 0.18, 0.28, 0.38 and 0.48 percent of potassium. One group of eight male and one group of eight female poults were assigned to each of the sixteen treatments. Fecal samples were collected from each group during the last week of the experiment. These samples were dried and finely ground; potassium levels were determined by the method followed by Hassan and Knudsen (1968). Sodium Experiments. Experiment 3. In this experiment, a corn-soybean meal type basal diet was used. The composition of this diet is presented in Table 2. The sodium content of the basal diet was 0.06 percent. The basal diet was supplemented with graded levels of 0.00, 0.04, 0.08 and 0.12 percent of sodium and 125, 175, 350 and 700 I.C. units of vitamin D 3 per kg. of diet. Nine male and nine female poults were assigned to each of the sixteen treatments.
Ground yellow corn Soybean meal (50% protein) Dehy. alfalfa meal (17% protein) Corn oil CaHP04H20 Calcium carbonate MnS04H20 ZnS0 4 -7H 2 0 FeC 6 H 2 0 5 -5H 2 0 Vitamin premix2 'Calculated averaged composition: Protein, % 28.3 Metabolizable energy, Kcal./kg. 2814 Calcium, % 1.80 Phosphorus, % 0.91 2 Vitamin premix provided the following per kilogram of diet: 5500 I.U. stablizied vitamin A, 22 I.U. stabilized vitamin E, 6.6 mg. riboflavin, 11 mg. calcium pantothenate, 66 mg. niacin, 22 meg. vitamin B l 2 , 1 gm. choline chloride, 1.5 g. methionine-hydroxy-analog, and 22 mg. penicillinstreptomycin (1:3).
Experiment 4. The same basal diet was used as in experiment 3. This diet was supplemented with 0.03, 0.07 and 0.11 percent of sodium; and 200, 400, 800 and 1600 I.C. units of supplemental vitamin D 3 per kg. of diet. One group of eight male and one group of eight female poults were assigned to each of the twelve treatments. Fecal samples were prepared in the same manner as in the second potassium experiment. Blood samples from three birds in each group were taken by cardiac puncture at the end of the experiment and were analyzed for sodium level in serum. Magnesium Experiments. Experiment 5. The effect of vitamin D 3 on magnesium level in serum and tibiae of four-week old turkeys was studied. Four-week body weight data were also obtained in this study. Composition of basal diet is presented in Table 1. According to chemical analysis the unsupplemented
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Potassium Experiments. Experiment 1. A purified diet was utilized; the composition of this diet is presented in Table 1. Chemical analysis indicated that the unsupplemented basal diet contained 0.055 percent potassium. One group of eight male and one group of eight female poults were assigned to each of the sixteen dietary treatments. The levels of supplemental vitamin D 3 were 400, 800, 1600 and 3200 I.C. units per kg. of diet. Supplemental potassium levels were 0.08, 0.18, 0.28 and 0.38 percent. Because of the high mortality in poults receiving the lowest level of potassium, four-week body weight data of these groups were not subjected to analysis of variance.
Per kg. of diet g390.0 489.0 30.0 30.0 20.0 34.3 0.2 0.2 0.2 6.2
485
VITAMIN D , X K, NA AND MG IN POULT DIETS
TABLE 3.—Body weight, feed efficiency and survival data from experiment 1 Vitamin D 3 I.C. units/kg.
Avg. body wt. at 4 weeks, g.1
Feed/gain, 0-4 weeks
400
800
K levels, % 0.24 0.34 0.44 Avg.
389 271 419
447 497 405
360a
0.14 0.24 0.34 0.44 Avg.
1.77 1.71 2.04 1.52 1.76
3200
Avg.
428 470 513
436 550 525
425a 447ab 466b
450b
470bc
504c
1.68 1.47 1.38 1.41 1.48
1.88 1.57 1.42 1.46 1.58
1.39 1.38 1.39 1.32 1.37
1600
1.68 1.53 1.56 1.43
0.14 0.24 0.34 0.44 Avg.
basal diet contained 405 p.p.m. of magnesium. Distilled water was furnished ad libitum for the last three weeks of the experiment. The basal diet was supplemented with 0, 50, 150 and 350 p.p.m. of magnesium. Levels of vitamin D 3 added to the basal diet were 600, 2400 and 4800 I.C. units per kg. Eight male and eight female poults were assigned to each of the twelve treatments. Blood and tibia samples were taken from three and four birds in each group, respectively, at the termination of the experiment. Experiment 6. The same basal diet was used as in experiment 5. Levels of supplemental magnesium were 0,100, and 300p.p.m.; levels of vitamin D 3 were 600, 2400 and 4800 I.C. units per kg. Eight female and eight male poults were assigned to each of the nine treatments. RESULTS AND DISCUSSION Experiment 1. Data concerning body weight gain, efficiency of feed utilization and sur-
vival of poults in this experiment are presented in Table 3. High mortality of poults fed the lowest (0.14%) level of potassium indicated that this level was too low for survival, regardless of vitamin D 3 levels. Because of poor survival body weight gain data for poults receiving the lowest level of potassium were not included in the analysis of variance. Body weight gain was significantly lower in poults fed the lowest level (400 I.C. units/kg.) of vitamin D 3 than in poults receiving the higher levels. Poults receiving 0.44 percent of dietary potassium were significantly heavier at four weeks of age than those receiving 0.24 percent potassium. The efficiency of feed utilization and survival increased when vitamin D 3 and potassium levels were increased in the diet. The results of this experiment indicated that vitamin D 3 influenced body weight gain by affecting either directly or indirectly, potassium metabolism, bone growth or some other metabolic processes. However, it is difficult to determine to what extent vitamin D 3 influenced potassium utilization, because
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34 56 25 38 19 62 62 88 88 75 70 44 75 81 81 100 88 89 69 100 69 72 78 50 1 Each individual body weight value is the mean for survivors in two replicate groups of eight birds each. For either vitamin D 3 or potassium levels, composite average values followed by the same letter are not significantly different from each other (P < 0.05). Survival to 4 weeks, %
486
A. AKSOY AND T. W .
SULLIVAN
TABLE 4.—Body weight, feed efficiency, survival and fecal potassium data from experiment 2
600
Vitamin D 3 ,, I.C. units/kg. 1200 1800
Avg.
339 445 398 434
365 401 405 494
336a 393b 382b 430c
391b
404bc
426c
1.75 2.05 1.92 1.45 1.79
1.68 1.55 1.56 1.60 1.60
1.80 1.48 1.58 1.60 1.62
1.64 1.61 1.53 1.50 1.57
1.72 1.67 1.65 1.54
0.24 0.34 0.44 0.54 Avg.
75 69 81 88 78
88 94 88 88 90
62 81 88 100 83
81 88 100 88 89
76 83 89 91
0.24 0.34 0.44 0.54 Avg.
6560 6000 8790 11640 8248
4000 5570 8980 10100 7162
3980 8180 9960 11980 8525
2520 5920 7610 7420 5868
4265 6418 8835 10285
307 328 323 362
334 399 400 430
330a
Feed/gain, 0-4 weeks
0.24 0.34 0.44 0.54 Avg.
Survival to 4 weeks, %
Fecal K, p.p.m.2
Avg. body wt. at 4 weeks, g.1
1 Each individual body weight value is the mean for survivors in two replicate groups of eight birds each. For either vitamin D 3 or potassium levels, composite average values followed by the same letter are not significantly different from each other (P < 0.05). 2 Each individual fecal potassium value is the mean for two determinations per treatment.
TABLE 5.—Body weight and feed efficiency data from experiment 3 Vitamin D 3 , I.C. units/kg.
Avg. body wt. at 4 weeks, g.1
Feed/gain 0-4 weeks
1
700
Avg.
202 338 506 496
197 384 524 517
186a 348b 478c 473c
369b
386bc
406c
4.20 2.18 1.78 1.80 2.49
4.24 2.26 1.98 1.60 2.52
3.92 2.17 1.78 1.72 2.40
125
175
N a levels, % 0.06 0.10 0.14 0.18 Avg.
168 309 422 398
177 359 461 480
324a
0.06 0.10 0.14 0.18 Avg.
5.92 2.38 2.05 1.95 3.08
350
4.57 2.25 1.90 1.77
Each individual body weight value is the mean for survivors in two replicate groups of nine birds each. For either vitamin D 3 or Na levels, composite average values followed by the same letter are not significantly different from each other (P < 0.05).
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3600
K levels, % 0.24 0.34 0.44 0.54 Avg.
487
VITAMIN D 3 X K, NA AND MG IN POULT DIETS
TABLE 6.—Body weight, feed efficiency and serum and fecal sodium data from experiment 4 Vitamin D 3 ,, I.C. units/kg.
200 Avg. body wt. at 4 weeks, g.1
Feed/gain, 0-4 weeks
N a levels, % 0.09 0.13 0.17 Avg. 0.09 0.13 0.17
400
800
1600
Avg. 285a 443b 491c
247 405 454
286 432 481
329 430 505
278 505 525
369a
400ab
421 be
436c
2.44 2.16 1.72 2.11
2.08 1.92 1.70 1.90
2.44 1.72 1.62 1.93
2.44 2.00 1.70
Serum Na, p.p.m.2
0.09 0.13 0.17 Avg.
3271 3175 3275 3240
3162 3283 3233 3226
3083 3340 3258 3230
3042 3250 3425 3239
3140a 3264b 3298b
Fecal Na, p.p.m.3
0.09 0.13 0.17 Avg.
2820 3760 6040 4207
3190 3620 4540 3783
2800 4090 5120 4003
2630 4040 4700 3790
2860 3878 5100
1
Each individual body weight value is the mean for survivors in two replicate groups of eight birds each. For either vitamin D3 or Na levels, composite average values followed by the same letter are not significantly different from each other (P < 0.05). 2 Each individual serum sodium value is the mean for six samples, three from each replicate group assigned to each treatment. 3 Each individual fecal sodium value is the mean for two determinations per treatment.
of the very close relationship of vitamin D with calcium utilization and metabolism. Experiment 2. There was a significant (P < 0.05) D 3 x potassium effect on four-week body weight gain. There were highly significant differences in body weights of poults due to both vitamin D 3 and potassium levels. The lowest level of potassium and the lowest level of vitamin D 3 each had depressing effects on body weight gains to four weeks of age (Table 4). The higher levels of vitamin D 3 tended to reduce fecal excretion of potassium (Table 4). Poults fed 1800 I.C. units of vitamin D 3 did not follow this trend, however. Fecal potassium levels steadily increased when dietary potassium levels were increased. Vitamin D may act on the tubular function of kidney by decreasing excretion of potassi-
um or on transfer of the ion from one tissue to another. The latter possibility was not shown in the experiment conducted by Wasserman (1962). Wasserman also found that vitamin D 3 did not significantly increase the duodenal absorption of potassium. The efficiency of feed utilization increased when dietary potassium and vitamin D 3 levels increased (Table 4). Survival percentage was the least with the lowest level of either potassium or vitamin D 3 . Experiment 3. The D 3 x sodium effect on body weight gain was not statistically significant. However, there were highly significant (P < 0.01) differences in body weight among vitamin D 3 levels and among sodium levels. Duncan's multiple range test showed that body weight gain was significantly depressed in poults fed the lowest level of sodium and
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Avg.
2.79 2.20 1.75 2.25
488
A. AKSOY AND T. W. SULLIVAN TABLE 7.-
-Body weight, feed efficiency, survival and tibiae and serum magnesium data from experiment 5 600
Vitamin D 3 , I.C.units/kg. 2400 4800
Avg.
Mg., p.p.m. Avg. body wt. at 4 weeks, g.'
Feed/gain, 0-4 weeks
405 453 555 755
392 398 433 443
482 448 453 393
477 451 512 510
Avg.
416a
469b
488b
405 455 555 755
1.60 1.59 1.56 1.49 1.56
1.48 1.46 1.55 1.40 1.47
1.52 1.54 1.46 1.37 1.47
1.53 1.53 1.52 1.42
88 75 94 94 88
94 100 94 100 97
94 100 100 100 98
92 92 96 98
4350 5361 6460 6855 5756a
4860 5396 6446 6371 5768a
4965 6022 6678 6596 6065b
4725a 5593b 6528c 6608c
19.7 34.2 33.9 37.0 31.2
27.9 23.6 28.1 33.5 28.3
23.9 28.1 26.6 31.5 27.8
23.8a 28.6ab 29.9b 34.0b
405 455 555 755
Survival to 4 weeks, %
Avg. 2
Tibia Mg, p.p.m.
405 455 555 755 Avg.
Blood serum Mg., p.p.m.3
405 455 555 755 Avg.
1
Each individual body weight value is the mean for survivors in two replicate groups of eight birds each. For either vitamin D 3 or Mg levels, composite average values, followed by the same letter are not significantly different from each other (P < 0.05). 2 Each tibia Mg value is the mean of eight determinations, four samples from each of two replicate groups per treatment. 3 Each serum Mg value is the mean for six determinations, three samples from each of the two replicate groups per treatment. Levels of Mg in p.p.m. H- 10 = mg./lOO ml. or mg. %.
the lowest level of D 3 (Table 5). Poults receiving 700 I.C. units of vitamin D 3 were significantly heavier than those receiving 175 I.C. units per kg. of diet. The efficiency of feed utilization increased as dietary sodium and dietary vitamin D 3 levels were increased. Since mortality rates were very low and apparently not influenced by either D 3 or sodium levels, survival data from experiments 3 and 4 are not presented.
Experiment 4. Again, no significant interaction occurred between dietary sodium and vitamin D 3 relative to four-week body weight. Body weight gains of poults receiving 800 and 1600 I.C. units vitamin D 3 were significantly greater than the gain of poults receiving 200 I.C. units vitamin D 3 (Table 6). Each increment of dietary sodium was associated with a significant (P < 0.05) increase in body weight gain.
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Avg.
450ab 432a 466bc 482c
489
VITAMIN D 3 X K, NA AND MG IN POULT DIETS
No significant differences occurred among vitamin D 3 levels with regard to sodium in serum. This agrees with the results of Wasserman (1962) and Richardson and Welt (1965). The two higher levels of dietary sodium significantly (P < 0.05) increased the Na level in serum. Fecal sodium levels were increased with each increment of dietary Na.
Serum magnesium levels decreased as dietary vitamin D 3 levels increased. But these differences among vitamin D 3 levels were not significant. A depressing effect of vitamin
TABLE 8.—Body weight, feed efficiency and survival data from experiment 6 Vitamin D 3 , I.C. units/kg.
600
2400
4800
Avg.
405 505 705
340 380 329
412 432 493
470 511 480
407a 441b 431b
Avg.
346a
446b
487c
405 505 705
1.87 1.76 1.81 1.81
1.60 1.56 1.49 1.55
1.52 1.48 1.39 1.46
Mg., p.p.m. Avg. body wt. at 4 weeks, g.1
Feed/gain, 0-4 weeks
Avg.
44 71 88 81 94 56 75 75 38 100 73 81 Avg. 46 94 79 1 Each individual body weight value is the mean for survivors in two replicate groups of eight birds each. For either vitamin D 3 or Mg levels, composite average values followed by the same letter are not significantly different from each other (P < 0.05). Survival to 4 weeks, %
405 505 705
1.66 1.60 1.56
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Experiment 5. The D 3 x magnesium effect on body weight gain was not statistically significant. However, highly significant (P < 0.01) body weight differences were noted among dietary magnesium levels and vitamin D 3 levels. The body weight of birds receiving 755 p.p.m. of magnesium was significantly (P < 0.05) greater than that of birds receiving 405 and 455 p.p.m. (Table 7). Efficiency of feed utilization was similar in poults receiving the three lower levels of magnesium; however, feed efficiency was improved with 755 p.p.m. of magnesium. Neither magnesium nor vitamin D 3 levels greatly influenced the survival of poults in this experiment. Differences among magnesium and among vitamin D 3 levels relative to magnesium con-
centration in tibiae of four-week old poults were highly significant (P < 0.01). But there was no significant interaction between magnesium and vitamin D 3 with respect to tibiaemagnesium concentration. Tibia magnesium content in poults receiving 600 and 2400 I.C. units vitamin D 3 per kg. of diet was significantly less than in poults receiving 4800 I.C. units per kg. (Table 7). When dietary magnesium level was increased the magnesium level in tibiae increased accordingly. Our data support previous reports by Worker and Migicovsky (1961). These workers found that vitamin D 3 significantly increased deposition of 28 Mg in tibiae of chicks following administration of the isotope orally. When the isotope was administered subcutaneously, the level of magnesium in bone was depressed significantly. Vitamin D treatment caused a very rapid decrease in bone magnesium concentration in rats placed on a low magnesium diet (Lifshitz et al., 1967).
490
A. AKSOY AND T. W. SULLIVAN
Results obtained from both the present study and previous work suggest that vitamin D has some influence on the metabolism of magnesium. This influence may be at the absorption site (Meintzer and Steenbock, 1956; Wasserman, 1962) or on cellular transport which has been reported by many workers.
functions of vitamin D have been summarized by DeLuca (1971) and by Lawson and Emtage (1974). The vitamin is currently believed to function in calcium absorption and transport through an active cation-oriented process involving a calcium binding protein and/or a calcium ATPase. These functions are dynamically interrelated and involved in a complex system regulating plasma calcium and phosphate concentrations. Results from the present study agree with the current theory of vitamin D's metabolic functions. In the present study no evidence was obtained indicating a direct effect of vitamin D on either potassium, sodium or magnesium. However, it was apparent that vitamin D influenced the growth, feed utilization, and fecal and tissue contents of starting turkeys fed graded levels of potassium and magnesium. These effects were apparently indirect and were probably mediated through the vitamin's primary role in calcium and phosphorus metabolism. REFERENCES
Experiment 6. Highly significant (P < 0.01) and significant (P < 0.05) differences, respectively, were observed among vitamin D 3 and among magnesium levels relative to body weight. Poults receiving 405 p.p.m. of dietary magnesium gained significantly less than those receiving 505 and 705 p.p.m. (Table 8). Each increment of dietary vitamin D 3 significantly increased the four-week body weight gain. The effect of vitamin D 3 level on survival was much more pronounced than in the previous experiment. Overall survival rate was lower in this experiment than in the previous experiment. Two factors may have contributed to these differences. First, adifferent experimental design was used, and second, distilled water was provided during the entire experimental period of 28 days, whereas, distilled water was provided only the last 20 days of the first experiment. Recent research concerning the metabolic
DeLuca, H. F., 1971. Biological systems. The Vitamins, Edited by H. W. Sebrell and R. S. Harris. Academic Press, New York. Vol. 3: 240-247. Duncan, D. B., 1955. Multiple range and multiple F test. Biometrics, 11: 1-42. Hanna, S., 1961. Influence of large doses of vitamin D on magnesium in rats. Metabolism, 10: 735-743. Harrison, H. E., and H. C. Harrison, 1964. The interaction of vitamin D and parathyroid hormone in calcium, phosphorus and magnesium homeostasis in the rat. Metabolism, 13: 952-958. Hassan, N. A. K., and D. Knudsen, 1968. Methods of Chemical Analysis of Plant, Soil and Water. Univ. of Nebr. College of Agriculture, Soil, Plant, and Water Testing Service. Lawson, D. E. M., and J. S. Emtage, 1974. Molecular action of vitamin D in the chick intestine. Vitamins Hormones, 32: 277-298. Lifshitz, F., H. C. Harrison and H. E. Harrison, 1967. Effects of vitamin D on magnesium metabolism in rats. Endocrinology, 81: 849-853. McHargue, J. S., and W. R. Roy, 1930. Effect of ultra-violet irradiation on the magnesium content of rats receiving reflected sunlight and a uniform
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D 3 on serum magnesium level has been previously reported by several workers in various species of animals (rat, Hanna, 1961; rat, Richardson et ai, 1963; rat, Harrison and Harrison, 1964; rat, Richardson and Welt, 1965; dog, Wallach and Carter, 1961; dog, Wallach et ai, 1966). The depressing effect of vitamin D on serum magnesium and the stimulating effect on concentration in bone and certain other tissues have caused some speculation concerning the vitamin's function. It has been suggested that hypomagnesemia was due to a redistribution of magnesium from the extracellular phase to some other depots, such as bone (Richardson and Welt, 1965).
491
VITAMIN D 3 X K, N A AND MG IN POULT DIETS
stock ration. Am. J. Physiol. 92: 651-655. Meintzer, R. B., and J. Steenbock, 1956. Vitamin D and the magnesium absorption. J. Nutr. 56: 285-294. Richardson, J. A., W. D. Huffines and L. G. Welt, 1963. The effect of coincident hypercalcemia and potassium depletion on the rat kidney. Metabolism, 12: 560-569. Richardson, J. A., and L. G. Welt, 1965. The hypomagnesema of vitamin D administration. Proc. Soc. Exp. Biol. Med. 118: 512-514. Steel, R. G. D., and J. H. Torrie, 1960. Principles and Procedures of Statistics. McGraw-Hill Book Co., Inc., New York, Toronto, London. Wallach, S., and A. C. Carter, 1961. Metabolic and
renal effects of acute hypercalcemia in dogs. Am. J. Physiol. 200: 359-366. Wallach, S., J. V. Bellavia, J. Schorr and P. J. Camponia, 1966. Effect of vitamin D on the tissue distribution and transport of electrolytes 47Ca and 28 Mg. Endocrinology, 79: 773-782. Wasserman, R. H., 1962. Studies on vitamin D 3 and the intestinal absorption of calcium and other ions in the rachitic chick. J. Nutr. 77: 69-80. Worker, N. A., and B. B. Migicovsky, 1961. Effect of vitamin D on the utilization of beryllium, magnesium, calcium, strontium and barium in the chick. J. Nutr. 74: 409-494.
AYHAN AKSOY 3 AND T . W . SULLIVAN
Department of Poultry & Wildlife Sciences, University of Nebraska, Lincoln, Nebraska 68583 (Received for publication June 28, 1976)
ABSTRACT Three experiments were conducted with Large White turkeys to four weeks of age. The interrelationship of zinc and vitamin D 3 was studied in two experiments. Iron and vitamin D 3 were involved in one experiment. One group of ten male and one group of ten female poults were randomly assigned to each treatment within an experiment. All experiments involved a factorial arrangement of two dietary variables, D 3 and either zinc or iron. For example, all possible combinations of four zinc levels; 31, 46, 76 and 106 p.p.m. and three vitamin D 3 levels; 600, 1200, and 3600 I.C. units/kg. were fed in the second zinc experiment. Significant (P < 0.05) weight gain differences occurred among both zinc and D 3 levels in this experiment. Four week body weights were 356, 436, 459 and 444 g., respectively, for Zn levels; and 409, 410 and 452 g., respectively, for D 3 levels. Significant (P < 0.05) interactions occurred between dietary zinc and D 3 levels, and between iron and D 3 levels relative to body weight gain. Vitamin D 3 tended to increase tibia zinc levels in poults receiving higher zinc levels; the opposite occurred with lower levels of the element. Zinc level in feces was decreased with vitamin D 3 . Hemoglobin level was not influenced by D 3 but was increased by iron. POULTRY SCIENCE 56: 491-498, 1977
INTRODUCTION
T
HE literature shows considerable inconsistency and uncertainty as to the effect
of vitamin D on zinc metabolism. This might be due in part to the close relationships
between zinc and calcium, and between calcium and
vitamin
D. Wasserman
reported that duodenal absorption of
(1962) 64
Zn
in the chick was not enhanced by vitamin D3.
But, Martin and Patrick (1961) found
that addition of vitamin D 3 to a deficient diet decreased liver 65 Zn regardless of dietary
1. Published as paper number 5141, Journal Series, Nebraska Agricultural Experiment Station. 2. From a thesis submitted by the senior author in partial fulfillment of the requirements for the Ph.D. degree. 3. Present address: Animal Science Department, College of Agriculture, Ataturk University, Erzurum, Turkey.
Zn or Ca level. The presence of vitamin D 3 increased the
65
Zn level of the tibia when
the diet contained 1.0 percent calcium, and decreased the level in the presence of
1.6
percent calcium in both the zinc-adequate and deficient groups. A positive effect of vitamin D on zinc metabolism in the chicken was
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Interrelationship of Dietary Vitamin D 3 with Zinc and Iron in Young Turkeys 12
J. B. COOPER
REFERENCES
lowest for treatment 4, Table 1. Treatment 4 also resulted in the lowest number of squabs being produced per pair for any of the five treatments. Treatment 5 (open lofts)
was
second lowest. ACKNOWLEDGEMENTS The author is indebted to Dr. Walter E.
Levi, W. M., 1941. The Pigeon. R. L. Bryan Co., Columbia, S.C. Platt, C. S., and R. S. Dare, 1937. New Jersey State Pigeon Breeding Test. New Jersey Agri. Exp. Sta. Bui. 634. Steel, R. G. D„ and J. H. Torrie, 1960. Principles and Procedures of Statistics. McGraw-Hill, New York.
Johnston of the Experimental Statistical Services of Clemson University for the statistical analysis.
AYHAN A K S O Y 3 AND T . W . SULLIVAN
Department of Poultry & Wildlife Sciences, University of Nebraska, Lincoln, Nebraska 68583 (Received for publication June 28, 1976)
ABSTRACT Six experiments were conducted with Large White and Bronze turkeys to four weeks of age. The interrelationship of dietary vitamin D 3 with either potassium, sodium or magnesium was studied in two experiments each. One group of 8-9 male and one group of 8-9 female poults were randomly assigned to each treatment within an experiment. The design of all experiments involved a factorial arrangement of two variables: vitamin D 3 and a mineral element. For example, all possible combinations of four dietary potassium levels, 0.24, 0.34, 0.44, 0.54%; and four vitamin D 3 levels 400, 800, 1600 and 3200 I.C. units/kg. were fed in the second potassium experiment. Significant (P < 0.05) weight gain differences occurred among both K levels and D 3 levels in this particular experiment. Average four-week body weights were 336, 393, 382 and 430 g., respectively, for potassium levels and 330, 391, 404 and 426 g., respectively, for vitamin D 3 levels. There was a significant interaction between dietary K and D 3 levels relative to four-week body weight gains. Vitamin D 3 tended to increase retention of both potassium and sodium in the body. A significant interaction occurred between dietary magnesium and vitamin D 3 relative to body weight gain. Vitamin D 3 increased magnesium content in tibiae ash and decreased magnesium content in serum. POULTRY SCIENCE 56: 482-491, 1977
INTRODUCTION
T
HE effect of vitamin D on calcium metabolism has been extensively studied but
little is known about its effect on the metabo-
lism of other cations. A few studies have indicated that vitamin D plays a role in the utilization of other cations. As early as 1930 McHargue et al. reported that the carcass magnesium content of rats subjected to ultraviolet irradiation was less than that of control
1. Published as paper Number 5142, Journal Series, Nebraska Agricultural Experiment Station. 2. From a thesis submitted by the senior author in partial fulfillment of the requirements for the Ph.D. degree. 3. Present address: Animal Science Department, College of Agriculture, Ataturk University, Erzurum, Turkey.
animals.
Meintzer
and
Steenbock
(1956)
showed that vitamin D supplementation increased magnesium absorption in rats. Similar results were obtained by Wasserman (1962) with chicks. Worker and Migicovsky (1961) reported that vitamin D 3 significantly creased the deposition of
28
in-
Mg in the tibiae
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Interrelationship of Dietary Vitamin D 3 with Potassium, Sodium and Magnesium in Young Turkeys1-2
483
VITAMIN D 3 X K, NA AND MG IN POULT DIETS
EXPERIMENTAL PROCEDURE Six experiments were conducted with Large White and Bronze turkeys, day-old to four weeks of age. A factorial arrangement of dietary treatments, involving all combinations of three or four vitamin D 3 levels and three or four levels of the particular mineral element, was used in each experiment. All poults were maintained in thermostatically controlled, electrically heated battery brooders with raised wire floors throughout the experimental period. The experimental diet and water were supplied ad libitum. Determination of sodium in basal diet, feces and blood serum; potassium in basal diet and feces; magnesium in basal diet, blood serum and tibiae were made by methods described by Hassan and Knudsen (1968). Calcium and phosphorus levels in all basal diets were maintained somewhat higher than known requirements. It was hoped that moderate excesses of dietary Ca and P would minimize the effect of low D 3 levels on the absorption and metabolism of these elements and overall poult performance. All diets were
supplemented with penicillin-streptomycin (1:3) to provide 22 parts per million (p.p.m.) of the antibiotics. Four-week body weight data were subjected to analysis of variance (Steel and Torrie, 1960) and Duncan's (1955) multiple range test. Individual bird data were used in these statistical procedures. Feed efficiency and survival data were not analyzed statistically because true replicates were not involved in TABLE 1.—Composition of the basal diet for potassium and magnesium experiments' Ingredients
Per kg. of diet
Dextrose sugar (cerelose) Isolated soybean protein Solka Floe (wood flour) Corn oil Methionine-hydroxy-analog Glycine Ca3(P04)2 NaCl MgS04-7H202 FeC6H507-5H20 ZnS04-7H20 CuS04-5H20 KC1 3 MnS04H20 KI Choline chloride (50%) Myvamix (44 I.U. vit. E/g.) Vitamin A premix (5,000 I.U./g.) Vitamin B premix in sucrose 4 Penicillin-streptomycin 5
463.00 370.00 40.00 45.00 5.00 2.00 40.00 5.00 4.80 0.60 0.44 0.04 8.00 0.38 0.04 5.00 1.00 3.00 5.00 0.50
g-
'Calculated average composition: Protein, % 30.0 Metabolizable energy, Kcal./kg. 3046 Calcium, % 1.57 Phosphorus, % 1.05 2 Omitted from the basal diet in magnesium experiments. 3 Omitted from the basal diet in potassium experiments. "Vitamin B premix provided the following per kg. of diet: 0.02 mg. vitamin B12,0.30 mg. d-biotin, 4 mg. menadione, 8 mg. pyridoxine, 6 mg. folic acid, 10 mg. riboflavin, 30 mg. calcium pantothenate, 15 mg. thiamine HC1, and 100 mg. niacin. 5 Pro-Strep (Merck and Company) contains 44 mg. of penicillin-streptomycin, 1:3 per g.
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of chickens. Vitamin D treatment caused hypomagnesemia in rats (Hanna 1961; Richardson and Welt, 1965; Lifshitz et al., 1967). Vitamin D apparently affects the absorption and metabolism of potassium and sodium to a lesser extent than that of magnesium. Wasserman (1962) found that vitamin D 3 had no significant influence on absorption or tissue distribution of sodium and potassium in chicks with ligated duodenums. Large doses of vitamin D 2 given intramuscularly in rats had no influence on the sodium level of blood. But a lower plasma level of potassium was obtained following the administration of calciferol. The present investigation was undertaken to study possible interrelationships between dietary vitamin D 3 , sodium, potassium and magnesium in young turkeys.
484
A. AKSOY AND T. W. SULLIVAN
the experiments. Tissue and fecal composition data were evaluated by analysis of variance and Duncan's multiple range test.
TABLE 2.—Composition of basal diet for sodium experiments' Ingredients
Experiment 2. The basal diet, which was the same as used in experiment 1, was supplemented with 600, 1200, 1800 and 3600 I.C. units of vitamin D 3 per kg., and 0.18, 0.28, 0.38 and 0.48 percent of potassium. One group of eight male and one group of eight female poults were assigned to each of the sixteen treatments. Fecal samples were collected from each group during the last week of the experiment. These samples were dried and finely ground; potassium levels were determined by the method followed by Hassan and Knudsen (1968). Sodium Experiments. Experiment 3. In this experiment, a corn-soybean meal type basal diet was used. The composition of this diet is presented in Table 2. The sodium content of the basal diet was 0.06 percent. The basal diet was supplemented with graded levels of 0.00, 0.04, 0.08 and 0.12 percent of sodium and 125, 175, 350 and 700 I.C. units of vitamin D 3 per kg. of diet. Nine male and nine female poults were assigned to each of the sixteen treatments.
Ground yellow corn Soybean meal (50% protein) Dehy. alfalfa meal (17% protein) Corn oil CaHP04H20 Calcium carbonate MnS04H20 ZnS0 4 -7H 2 0 FeC 6 H 2 0 5 -5H 2 0 Vitamin premix2 'Calculated averaged composition: Protein, % 28.3 Metabolizable energy, Kcal./kg. 2814 Calcium, % 1.80 Phosphorus, % 0.91 2 Vitamin premix provided the following per kilogram of diet: 5500 I.U. stablizied vitamin A, 22 I.U. stabilized vitamin E, 6.6 mg. riboflavin, 11 mg. calcium pantothenate, 66 mg. niacin, 22 meg. vitamin B l 2 , 1 gm. choline chloride, 1.5 g. methionine-hydroxy-analog, and 22 mg. penicillinstreptomycin (1:3).
Experiment 4. The same basal diet was used as in experiment 3. This diet was supplemented with 0.03, 0.07 and 0.11 percent of sodium; and 200, 400, 800 and 1600 I.C. units of supplemental vitamin D 3 per kg. of diet. One group of eight male and one group of eight female poults were assigned to each of the twelve treatments. Fecal samples were prepared in the same manner as in the second potassium experiment. Blood samples from three birds in each group were taken by cardiac puncture at the end of the experiment and were analyzed for sodium level in serum. Magnesium Experiments. Experiment 5. The effect of vitamin D 3 on magnesium level in serum and tibiae of four-week old turkeys was studied. Four-week body weight data were also obtained in this study. Composition of basal diet is presented in Table 1. According to chemical analysis the unsupplemented
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Potassium Experiments. Experiment 1. A purified diet was utilized; the composition of this diet is presented in Table 1. Chemical analysis indicated that the unsupplemented basal diet contained 0.055 percent potassium. One group of eight male and one group of eight female poults were assigned to each of the sixteen dietary treatments. The levels of supplemental vitamin D 3 were 400, 800, 1600 and 3200 I.C. units per kg. of diet. Supplemental potassium levels were 0.08, 0.18, 0.28 and 0.38 percent. Because of the high mortality in poults receiving the lowest level of potassium, four-week body weight data of these groups were not subjected to analysis of variance.
Per kg. of diet g390.0 489.0 30.0 30.0 20.0 34.3 0.2 0.2 0.2 6.2
485
VITAMIN D , X K, NA AND MG IN POULT DIETS
TABLE 3.—Body weight, feed efficiency and survival data from experiment 1 Vitamin D 3 I.C. units/kg.
Avg. body wt. at 4 weeks, g.1
Feed/gain, 0-4 weeks
400
800
K levels, % 0.24 0.34 0.44 Avg.
389 271 419
447 497 405
360a
0.14 0.24 0.34 0.44 Avg.
1.77 1.71 2.04 1.52 1.76
3200
Avg.
428 470 513
436 550 525
425a 447ab 466b
450b
470bc
504c
1.68 1.47 1.38 1.41 1.48
1.88 1.57 1.42 1.46 1.58
1.39 1.38 1.39 1.32 1.37
1600
1.68 1.53 1.56 1.43
0.14 0.24 0.34 0.44 Avg.
basal diet contained 405 p.p.m. of magnesium. Distilled water was furnished ad libitum for the last three weeks of the experiment. The basal diet was supplemented with 0, 50, 150 and 350 p.p.m. of magnesium. Levels of vitamin D 3 added to the basal diet were 600, 2400 and 4800 I.C. units per kg. Eight male and eight female poults were assigned to each of the twelve treatments. Blood and tibia samples were taken from three and four birds in each group, respectively, at the termination of the experiment. Experiment 6. The same basal diet was used as in experiment 5. Levels of supplemental magnesium were 0,100, and 300p.p.m.; levels of vitamin D 3 were 600, 2400 and 4800 I.C. units per kg. Eight female and eight male poults were assigned to each of the nine treatments. RESULTS AND DISCUSSION Experiment 1. Data concerning body weight gain, efficiency of feed utilization and sur-
vival of poults in this experiment are presented in Table 3. High mortality of poults fed the lowest (0.14%) level of potassium indicated that this level was too low for survival, regardless of vitamin D 3 levels. Because of poor survival body weight gain data for poults receiving the lowest level of potassium were not included in the analysis of variance. Body weight gain was significantly lower in poults fed the lowest level (400 I.C. units/kg.) of vitamin D 3 than in poults receiving the higher levels. Poults receiving 0.44 percent of dietary potassium were significantly heavier at four weeks of age than those receiving 0.24 percent potassium. The efficiency of feed utilization and survival increased when vitamin D 3 and potassium levels were increased in the diet. The results of this experiment indicated that vitamin D 3 influenced body weight gain by affecting either directly or indirectly, potassium metabolism, bone growth or some other metabolic processes. However, it is difficult to determine to what extent vitamin D 3 influenced potassium utilization, because
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34 56 25 38 19 62 62 88 88 75 70 44 75 81 81 100 88 89 69 100 69 72 78 50 1 Each individual body weight value is the mean for survivors in two replicate groups of eight birds each. For either vitamin D 3 or potassium levels, composite average values followed by the same letter are not significantly different from each other (P < 0.05). Survival to 4 weeks, %
486
A. AKSOY AND T. W .
SULLIVAN
TABLE 4.—Body weight, feed efficiency, survival and fecal potassium data from experiment 2
600
Vitamin D 3 ,, I.C. units/kg. 1200 1800
Avg.
339 445 398 434
365 401 405 494
336a 393b 382b 430c
391b
404bc
426c
1.75 2.05 1.92 1.45 1.79
1.68 1.55 1.56 1.60 1.60
1.80 1.48 1.58 1.60 1.62
1.64 1.61 1.53 1.50 1.57
1.72 1.67 1.65 1.54
0.24 0.34 0.44 0.54 Avg.
75 69 81 88 78
88 94 88 88 90
62 81 88 100 83
81 88 100 88 89
76 83 89 91
0.24 0.34 0.44 0.54 Avg.
6560 6000 8790 11640 8248
4000 5570 8980 10100 7162
3980 8180 9960 11980 8525
2520 5920 7610 7420 5868
4265 6418 8835 10285
307 328 323 362
334 399 400 430
330a
Feed/gain, 0-4 weeks
0.24 0.34 0.44 0.54 Avg.
Survival to 4 weeks, %
Fecal K, p.p.m.2
Avg. body wt. at 4 weeks, g.1
1 Each individual body weight value is the mean for survivors in two replicate groups of eight birds each. For either vitamin D 3 or potassium levels, composite average values followed by the same letter are not significantly different from each other (P < 0.05). 2 Each individual fecal potassium value is the mean for two determinations per treatment.
TABLE 5.—Body weight and feed efficiency data from experiment 3 Vitamin D 3 , I.C. units/kg.
Avg. body wt. at 4 weeks, g.1
Feed/gain 0-4 weeks
1
700
Avg.
202 338 506 496
197 384 524 517
186a 348b 478c 473c
369b
386bc
406c
4.20 2.18 1.78 1.80 2.49
4.24 2.26 1.98 1.60 2.52
3.92 2.17 1.78 1.72 2.40
125
175
N a levels, % 0.06 0.10 0.14 0.18 Avg.
168 309 422 398
177 359 461 480
324a
0.06 0.10 0.14 0.18 Avg.
5.92 2.38 2.05 1.95 3.08
350
4.57 2.25 1.90 1.77
Each individual body weight value is the mean for survivors in two replicate groups of nine birds each. For either vitamin D 3 or Na levels, composite average values followed by the same letter are not significantly different from each other (P < 0.05).
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3600
K levels, % 0.24 0.34 0.44 0.54 Avg.
487
VITAMIN D 3 X K, NA AND MG IN POULT DIETS
TABLE 6.—Body weight, feed efficiency and serum and fecal sodium data from experiment 4 Vitamin D 3 ,, I.C. units/kg.
200 Avg. body wt. at 4 weeks, g.1
Feed/gain, 0-4 weeks
N a levels, % 0.09 0.13 0.17 Avg. 0.09 0.13 0.17
400
800
1600
Avg. 285a 443b 491c
247 405 454
286 432 481
329 430 505
278 505 525
369a
400ab
421 be
436c
2.44 2.16 1.72 2.11
2.08 1.92 1.70 1.90
2.44 1.72 1.62 1.93
2.44 2.00 1.70
Serum Na, p.p.m.2
0.09 0.13 0.17 Avg.
3271 3175 3275 3240
3162 3283 3233 3226
3083 3340 3258 3230
3042 3250 3425 3239
3140a 3264b 3298b
Fecal Na, p.p.m.3
0.09 0.13 0.17 Avg.
2820 3760 6040 4207
3190 3620 4540 3783
2800 4090 5120 4003
2630 4040 4700 3790
2860 3878 5100
1
Each individual body weight value is the mean for survivors in two replicate groups of eight birds each. For either vitamin D3 or Na levels, composite average values followed by the same letter are not significantly different from each other (P < 0.05). 2 Each individual serum sodium value is the mean for six samples, three from each replicate group assigned to each treatment. 3 Each individual fecal sodium value is the mean for two determinations per treatment.
of the very close relationship of vitamin D with calcium utilization and metabolism. Experiment 2. There was a significant (P < 0.05) D 3 x potassium effect on four-week body weight gain. There were highly significant differences in body weights of poults due to both vitamin D 3 and potassium levels. The lowest level of potassium and the lowest level of vitamin D 3 each had depressing effects on body weight gains to four weeks of age (Table 4). The higher levels of vitamin D 3 tended to reduce fecal excretion of potassium (Table 4). Poults fed 1800 I.C. units of vitamin D 3 did not follow this trend, however. Fecal potassium levels steadily increased when dietary potassium levels were increased. Vitamin D may act on the tubular function of kidney by decreasing excretion of potassi-
um or on transfer of the ion from one tissue to another. The latter possibility was not shown in the experiment conducted by Wasserman (1962). Wasserman also found that vitamin D 3 did not significantly increase the duodenal absorption of potassium. The efficiency of feed utilization increased when dietary potassium and vitamin D 3 levels increased (Table 4). Survival percentage was the least with the lowest level of either potassium or vitamin D 3 . Experiment 3. The D 3 x sodium effect on body weight gain was not statistically significant. However, there were highly significant (P < 0.01) differences in body weight among vitamin D 3 levels and among sodium levels. Duncan's multiple range test showed that body weight gain was significantly depressed in poults fed the lowest level of sodium and
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Avg.
2.79 2.20 1.75 2.25
488
A. AKSOY AND T. W. SULLIVAN TABLE 7.-
-Body weight, feed efficiency, survival and tibiae and serum magnesium data from experiment 5 600
Vitamin D 3 , I.C.units/kg. 2400 4800
Avg.
Mg., p.p.m. Avg. body wt. at 4 weeks, g.'
Feed/gain, 0-4 weeks
405 453 555 755
392 398 433 443
482 448 453 393
477 451 512 510
Avg.
416a
469b
488b
405 455 555 755
1.60 1.59 1.56 1.49 1.56
1.48 1.46 1.55 1.40 1.47
1.52 1.54 1.46 1.37 1.47
1.53 1.53 1.52 1.42
88 75 94 94 88
94 100 94 100 97
94 100 100 100 98
92 92 96 98
4350 5361 6460 6855 5756a
4860 5396 6446 6371 5768a
4965 6022 6678 6596 6065b
4725a 5593b 6528c 6608c
19.7 34.2 33.9 37.0 31.2
27.9 23.6 28.1 33.5 28.3
23.9 28.1 26.6 31.5 27.8
23.8a 28.6ab 29.9b 34.0b
405 455 555 755
Survival to 4 weeks, %
Avg. 2
Tibia Mg, p.p.m.
405 455 555 755 Avg.
Blood serum Mg., p.p.m.3
405 455 555 755 Avg.
1
Each individual body weight value is the mean for survivors in two replicate groups of eight birds each. For either vitamin D 3 or Mg levels, composite average values, followed by the same letter are not significantly different from each other (P < 0.05). 2 Each tibia Mg value is the mean of eight determinations, four samples from each of two replicate groups per treatment. 3 Each serum Mg value is the mean for six determinations, three samples from each of the two replicate groups per treatment. Levels of Mg in p.p.m. H- 10 = mg./lOO ml. or mg. %.
the lowest level of D 3 (Table 5). Poults receiving 700 I.C. units of vitamin D 3 were significantly heavier than those receiving 175 I.C. units per kg. of diet. The efficiency of feed utilization increased as dietary sodium and dietary vitamin D 3 levels were increased. Since mortality rates were very low and apparently not influenced by either D 3 or sodium levels, survival data from experiments 3 and 4 are not presented.
Experiment 4. Again, no significant interaction occurred between dietary sodium and vitamin D 3 relative to four-week body weight. Body weight gains of poults receiving 800 and 1600 I.C. units vitamin D 3 were significantly greater than the gain of poults receiving 200 I.C. units vitamin D 3 (Table 6). Each increment of dietary sodium was associated with a significant (P < 0.05) increase in body weight gain.
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Avg.
450ab 432a 466bc 482c
489
VITAMIN D 3 X K, NA AND MG IN POULT DIETS
No significant differences occurred among vitamin D 3 levels with regard to sodium in serum. This agrees with the results of Wasserman (1962) and Richardson and Welt (1965). The two higher levels of dietary sodium significantly (P < 0.05) increased the Na level in serum. Fecal sodium levels were increased with each increment of dietary Na.
Serum magnesium levels decreased as dietary vitamin D 3 levels increased. But these differences among vitamin D 3 levels were not significant. A depressing effect of vitamin
TABLE 8.—Body weight, feed efficiency and survival data from experiment 6 Vitamin D 3 , I.C. units/kg.
600
2400
4800
Avg.
405 505 705
340 380 329
412 432 493
470 511 480
407a 441b 431b
Avg.
346a
446b
487c
405 505 705
1.87 1.76 1.81 1.81
1.60 1.56 1.49 1.55
1.52 1.48 1.39 1.46
Mg., p.p.m. Avg. body wt. at 4 weeks, g.1
Feed/gain, 0-4 weeks
Avg.
44 71 88 81 94 56 75 75 38 100 73 81 Avg. 46 94 79 1 Each individual body weight value is the mean for survivors in two replicate groups of eight birds each. For either vitamin D 3 or Mg levels, composite average values followed by the same letter are not significantly different from each other (P < 0.05). Survival to 4 weeks, %
405 505 705
1.66 1.60 1.56
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Experiment 5. The D 3 x magnesium effect on body weight gain was not statistically significant. However, highly significant (P < 0.01) body weight differences were noted among dietary magnesium levels and vitamin D 3 levels. The body weight of birds receiving 755 p.p.m. of magnesium was significantly (P < 0.05) greater than that of birds receiving 405 and 455 p.p.m. (Table 7). Efficiency of feed utilization was similar in poults receiving the three lower levels of magnesium; however, feed efficiency was improved with 755 p.p.m. of magnesium. Neither magnesium nor vitamin D 3 levels greatly influenced the survival of poults in this experiment. Differences among magnesium and among vitamin D 3 levels relative to magnesium con-
centration in tibiae of four-week old poults were highly significant (P < 0.01). But there was no significant interaction between magnesium and vitamin D 3 with respect to tibiaemagnesium concentration. Tibia magnesium content in poults receiving 600 and 2400 I.C. units vitamin D 3 per kg. of diet was significantly less than in poults receiving 4800 I.C. units per kg. (Table 7). When dietary magnesium level was increased the magnesium level in tibiae increased accordingly. Our data support previous reports by Worker and Migicovsky (1961). These workers found that vitamin D 3 significantly increased deposition of 28 Mg in tibiae of chicks following administration of the isotope orally. When the isotope was administered subcutaneously, the level of magnesium in bone was depressed significantly. Vitamin D treatment caused a very rapid decrease in bone magnesium concentration in rats placed on a low magnesium diet (Lifshitz et al., 1967).
490
A. AKSOY AND T. W. SULLIVAN
Results obtained from both the present study and previous work suggest that vitamin D has some influence on the metabolism of magnesium. This influence may be at the absorption site (Meintzer and Steenbock, 1956; Wasserman, 1962) or on cellular transport which has been reported by many workers.
functions of vitamin D have been summarized by DeLuca (1971) and by Lawson and Emtage (1974). The vitamin is currently believed to function in calcium absorption and transport through an active cation-oriented process involving a calcium binding protein and/or a calcium ATPase. These functions are dynamically interrelated and involved in a complex system regulating plasma calcium and phosphate concentrations. Results from the present study agree with the current theory of vitamin D's metabolic functions. In the present study no evidence was obtained indicating a direct effect of vitamin D on either potassium, sodium or magnesium. However, it was apparent that vitamin D influenced the growth, feed utilization, and fecal and tissue contents of starting turkeys fed graded levels of potassium and magnesium. These effects were apparently indirect and were probably mediated through the vitamin's primary role in calcium and phosphorus metabolism. REFERENCES
Experiment 6. Highly significant (P < 0.01) and significant (P < 0.05) differences, respectively, were observed among vitamin D 3 and among magnesium levels relative to body weight. Poults receiving 405 p.p.m. of dietary magnesium gained significantly less than those receiving 505 and 705 p.p.m. (Table 8). Each increment of dietary vitamin D 3 significantly increased the four-week body weight gain. The effect of vitamin D 3 level on survival was much more pronounced than in the previous experiment. Overall survival rate was lower in this experiment than in the previous experiment. Two factors may have contributed to these differences. First, adifferent experimental design was used, and second, distilled water was provided during the entire experimental period of 28 days, whereas, distilled water was provided only the last 20 days of the first experiment. Recent research concerning the metabolic
DeLuca, H. F., 1971. Biological systems. The Vitamins, Edited by H. W. Sebrell and R. S. Harris. Academic Press, New York. Vol. 3: 240-247. Duncan, D. B., 1955. Multiple range and multiple F test. Biometrics, 11: 1-42. Hanna, S., 1961. Influence of large doses of vitamin D on magnesium in rats. Metabolism, 10: 735-743. Harrison, H. E., and H. C. Harrison, 1964. The interaction of vitamin D and parathyroid hormone in calcium, phosphorus and magnesium homeostasis in the rat. Metabolism, 13: 952-958. Hassan, N. A. K., and D. Knudsen, 1968. Methods of Chemical Analysis of Plant, Soil and Water. Univ. of Nebr. College of Agriculture, Soil, Plant, and Water Testing Service. Lawson, D. E. M., and J. S. Emtage, 1974. Molecular action of vitamin D in the chick intestine. Vitamins Hormones, 32: 277-298. Lifshitz, F., H. C. Harrison and H. E. Harrison, 1967. Effects of vitamin D on magnesium metabolism in rats. Endocrinology, 81: 849-853. McHargue, J. S., and W. R. Roy, 1930. Effect of ultra-violet irradiation on the magnesium content of rats receiving reflected sunlight and a uniform
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D 3 on serum magnesium level has been previously reported by several workers in various species of animals (rat, Hanna, 1961; rat, Richardson et ai, 1963; rat, Harrison and Harrison, 1964; rat, Richardson and Welt, 1965; dog, Wallach and Carter, 1961; dog, Wallach et ai, 1966). The depressing effect of vitamin D on serum magnesium and the stimulating effect on concentration in bone and certain other tissues have caused some speculation concerning the vitamin's function. It has been suggested that hypomagnesemia was due to a redistribution of magnesium from the extracellular phase to some other depots, such as bone (Richardson and Welt, 1965).
491
VITAMIN D 3 X K, N A AND MG IN POULT DIETS
stock ration. Am. J. Physiol. 92: 651-655. Meintzer, R. B., and J. Steenbock, 1956. Vitamin D and the magnesium absorption. J. Nutr. 56: 285-294. Richardson, J. A., W. D. Huffines and L. G. Welt, 1963. The effect of coincident hypercalcemia and potassium depletion on the rat kidney. Metabolism, 12: 560-569. Richardson, J. A., and L. G. Welt, 1965. The hypomagnesema of vitamin D administration. Proc. Soc. Exp. Biol. Med. 118: 512-514. Steel, R. G. D., and J. H. Torrie, 1960. Principles and Procedures of Statistics. McGraw-Hill Book Co., Inc., New York, Toronto, London. Wallach, S., and A. C. Carter, 1961. Metabolic and
renal effects of acute hypercalcemia in dogs. Am. J. Physiol. 200: 359-366. Wallach, S., J. V. Bellavia, J. Schorr and P. J. Camponia, 1966. Effect of vitamin D on the tissue distribution and transport of electrolytes 47Ca and 28 Mg. Endocrinology, 79: 773-782. Wasserman, R. H., 1962. Studies on vitamin D 3 and the intestinal absorption of calcium and other ions in the rachitic chick. J. Nutr. 77: 69-80. Worker, N. A., and B. B. Migicovsky, 1961. Effect of vitamin D on the utilization of beryllium, magnesium, calcium, strontium and barium in the chick. J. Nutr. 74: 409-494.
AYHAN AKSOY 3 AND T . W . SULLIVAN
Department of Poultry & Wildlife Sciences, University of Nebraska, Lincoln, Nebraska 68583 (Received for publication June 28, 1976)
ABSTRACT Three experiments were conducted with Large White turkeys to four weeks of age. The interrelationship of zinc and vitamin D 3 was studied in two experiments. Iron and vitamin D 3 were involved in one experiment. One group of ten male and one group of ten female poults were randomly assigned to each treatment within an experiment. All experiments involved a factorial arrangement of two dietary variables, D 3 and either zinc or iron. For example, all possible combinations of four zinc levels; 31, 46, 76 and 106 p.p.m. and three vitamin D 3 levels; 600, 1200, and 3600 I.C. units/kg. were fed in the second zinc experiment. Significant (P < 0.05) weight gain differences occurred among both zinc and D 3 levels in this experiment. Four week body weights were 356, 436, 459 and 444 g., respectively, for Zn levels; and 409, 410 and 452 g., respectively, for D 3 levels. Significant (P < 0.05) interactions occurred between dietary zinc and D 3 levels, and between iron and D 3 levels relative to body weight gain. Vitamin D 3 tended to increase tibia zinc levels in poults receiving higher zinc levels; the opposite occurred with lower levels of the element. Zinc level in feces was decreased with vitamin D 3 . Hemoglobin level was not influenced by D 3 but was increased by iron. POULTRY SCIENCE 56: 491-498, 1977
INTRODUCTION
T
HE literature shows considerable inconsistency and uncertainty as to the effect
of vitamin D on zinc metabolism. This might be due in part to the close relationships
between zinc and calcium, and between calcium and
vitamin
D. Wasserman
reported that duodenal absorption of
(1962) 64
Zn
in the chick was not enhanced by vitamin D3.
But, Martin and Patrick (1961) found
that addition of vitamin D 3 to a deficient diet decreased liver 65 Zn regardless of dietary
1. Published as paper number 5141, Journal Series, Nebraska Agricultural Experiment Station. 2. From a thesis submitted by the senior author in partial fulfillment of the requirements for the Ph.D. degree. 3. Present address: Animal Science Department, College of Agriculture, Ataturk University, Erzurum, Turkey.
Zn or Ca level. The presence of vitamin D 3 increased the
65
Zn level of the tibia when
the diet contained 1.0 percent calcium, and decreased the level in the presence of
1.6
percent calcium in both the zinc-adequate and deficient groups. A positive effect of vitamin D on zinc metabolism in the chicken was
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Interrelationship of Dietary Vitamin D 3 with Zinc and Iron in Young Turkeys 12