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The Influence of Periodic Removal of Calcium Sources from Rations of Chickens on Growth and Egg Production
750
J . G. OsTRANDER, R . JORDAN, W . J . S T A D E L M A N , J . C. ROGLER AND G. E . VAIL ing results from a research. A report. Purdue University, Mimeo. Reiser, R., 1950. Fatty acid changes in egg yolk of hens on a fat-free and a cottonseed oil ration. J. Nutrition, 40: 429-440. Reiser, R., 1951. The biochemical conversion of conjugated dienoic and trienoic fatty acids. Arch. Biochem. Biophys. 32: 113-120. Riemenschneider, R. W., N. R. Ellis and H. W. Titus, 1938. The fat acids in the lecithin and glyceride fractions of egg yolk. J. Biol. Chem. 126: 255-263. Russell, W. C , M. W. Taylor and H. A. Walker, 1941. The intake and output of fat by the hen on low fat and normal ration. Poultry Sci. 20: 372378. Shorland, F. B., 1951. The fatty acid composition of egg-yolk lipids. New Zealand J. Sci. Technol. B33: 224-229.
The Influence of Periodic Removal of Calcium Sources from Rations of Chickens on Growth and Egg Production G. A. DONOVAN, W. C. SHERMAN, K. E. PRICE 1 AND J. H. HARE Chas. Pfizer & Co., Inc., Terre Haute, Indiana (Received for publication September 18, 1959)
T
HE calcium requirement of young chicks as determined by various workers ranges from 0.66 to 2.14 percent of the diet (Ewing, 1951). This author suggests that the minimum calcium requirement for growing chicks is about 0.70%, but to provide a margin of safety the diet should furnish about 1.0% as recommended by the National Research Council (1954). The value of antibiotics administered in feed or water in the treatment of diseases of poultry has been well documented. Recently it has been demonstrated that the in vitro antibacterial activity of antibiotics is impaired by salts of various metallic ions (Newton, 1953; Weinberg, 1
Present address: Bristol Laboratories, Syracuse,
N.Y.
1955; Donovick el al., 1948; Schoehhard and Stafseth, 1953). The relative susceptibility of 10 antibiotics to inhibition by salts of calcium was determined by Price et al. (1955). They found that streptomycin and neomycin were markedly inhibited, that the tetracycline antibiotics were substantially inhibited, but that the antibacterial action of penicillin and oleandomycin was not influenced by calcium. Apparently part of the same phenomenon is the finding that absorption of oxytetracycline2 from the intestine of the chick is substantially inhibited by calcium ions as shown in experiments with exposed ligated duodenal loops (Price el al., 1958). 2
Inc.
Terramycin, Trademark of Chas. Pfizer & Co.,
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depot fat of the fowl as affected by the ingestion of large amounts of different fats. Biochem. J. 28: 96S-977. Dixon, W. J., and F. J. Massey, Jr., 1957. Introduction to Statistical Analysis. New York, McGrawHill Book Company, Inc. Feigenbaum, A. S., and H. Fisher, 1959. The influence of dietary fat on the incorporation of fatty acids into the body and egg fat of the hen. Arch. Biochem. Biophys. 79: 302-306. Fisher, H., and G. A. Leveille, 1957. Observations on the cholesterol, linoleic and linolenic acid content of eggs as influenced by dietary fats. J. Nutrition, 63: 119-129. McCollum, E. V., J. G. Halpin and A. H. Drescher, 1912. Synthesis of lecithin in the hen and the character of the lecithins produced. J. Biol. Chem. 13: 219-224. Miles, S. R., 1957. Drawing conclusions and report-
751
PERIODIC REMOVAL OF CALCIUM
of the life cycle, simulating the periods during which high-level antibiotics might be applied to combat disease. Normal calcium levels were fed during other than the experimental periods. The females in this experiment were removed at 35 days due to lack of space when males were transferred to finishing batteries. Egg Production Study.—One experiment was conducted with S. C. White Leghorn hens eleven months old which had been in production about six months. Rations Ei and E2, Table 1, which provided approximately 0.12% and 1.3% calcium, were tested. There were two floor pen groups of 18 birds on each treatment. RESULTS
Growing Birds.—Results of experiments 1 to 4 on continuous feeding of low calcium from day of age to 28 days are presented in Table 2. The six low-calcium EXPERIMENTAL groups in this series received diets conGrowth Studies.—Five experiments were taining from 0.702% Ca down to 0.138%. conducted with Nichols X Vantress cross- In three of these six groups there was a breeds. In experiments 1 to 4, low calcium significant retardation of gain on low callevels were tested in comparison with cium during the first two weeks (expericalcium levels approximating National ment 2, 0.458% and 0.702% Ca; experiResearch Council (1954) requirements ment 4, 0.138% Ca). There were appreciwhen fed continuously from 0 to 28 days, able, but not significant, reductions in except that, in one trial, observations rate of gain in the other three low-calcium extended only to 25 days because of an groups in the 0-14 day period. That reoutbreak of Newcastle disease. Experi- ductions were not as severe in experiment mental diets are shown in Table 1. Obser- 3, which was essentially parallel to experivations were made of weight gains in the ment 2, is probably explained by the periods 0-14 days and 14-28 days, of generally slower rate of gain of all groups, feed utilization (lb. gain/lb. feed) 0-28 including the control, because of the presdays, and index of performance (gainX ence of Newcastle disease in experiment feed utilization) 0-28 days. Bone ash de- 3. terminations were made by the method of Especially noteworthy is the group in the A.O. A.C. (1955) on the second phalanx experiment 4 which received the extremely of the second digit of the right foot. The low calcium level of 0.138% for two weeks. fifth experiment was conducted to deter- At 13 to 14 days about a third of these mine the physiological effect of maintain- birds began to hobble around on their ing broiler chicks on sub-optimal calcium hocks and were unable to stand and walk for periods of 2 to 8 days at various stages normally so that, at this time, the dietary
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This action is reflected in enhancement of blood oxytetracycline levels with decreases of dietary calcium levels or with simultaneous introduction into the gut of compounds which bind calcium (Price et al, 1959a). From this it follows that high-level antibiotic administration via either the feed or drinking water for control of diseases should be more effective in the presence of low dietary calcium levels for short periods of time. Experimental evidence of this has been obtained by Price et al. (1959b). Before dietary calcium alteration is adopted in practice, it is essential to know whether a temporary reduction of the level of this mineral in the feed adversely affects growth or egg production. This question was investigated in the following experiments.
— _
— — _^_ 32.94 — 0.55 — — 1.90 — — — 0.40
0.47 1.100
936 21
•
0.46 0.702
24
0.46 0.458
1,080
24
100.
0.10 0.742
— 0.46 — 0.61 — 0.30
1,080
100.
0.30 0.10 0.742
—
— 0.60
— — 1.28 — 0.46 — 0.61 1.29
10.01
— — — — 38.85
47.04
B2
9.53
— — — — 38.62
48.36
Bi B,
0.46 0.930
24
1,080
100.
0.10 0.742
— 0.46 — 0.61 — 0.30
1.29
— 1.20
10.45
— —. — — 39.07
45.78
Broiler expt. 2 and 3
0.55 0.138
24
1,008
100.
0.742
0.55 1.138
24
1,008
100.
0.742
0.52
— • — 0.10
— 1.60 — 0.44
— — 0.10
—
9.06 2.63 2.63
— — — — 39.60
42.78
C»
0.44 0.52
•
— — 1.60
9.06 5.26
— .— — ~39.60
42.78
Ci
100.
0.742
0.52
— — 0.10
— 5.26 — 1.60 — 0.44
9.06
— — — — 39.60
42.78
a
0.55 2.138
24
1,008
Broiler expt. 4
Composition of rations (%)
0.55 0.20
990 22
100.
—
0.30 0.10 0.742
0.52
— — 0.44 1.60
4.90 2.63 0.18
— — — — 33.52
55.25
Di
0.55 0.60
990 22
100.
—
0.30 0.10 0.742
0.52
— — 0.44 1.60
4.90 1.58 1.23
— — — — 33.52
55.25
D2
Da
0.55 1.20
990 22
100.
—
0.30 0.10 0.742
0.52
— — 1.60 — 0.44
2.81
4.90
— — — — 33.52
55.25
Broiler expt. 5
0.50 0.119
968 17
100.
—
•
0.742
—
0.015
— — — — — 1.76 — 0.45
2.27
63.20 3.00 5.00 2.00 21.56
Ei
0.50 1.340«
949 17
100.
— —
0.742
0.015
— — — 0.45
2.13
—• — 1.83 2.27
61.00 3.00 5.00 2.00 21.56
Es
Layer expt.
Solka-floc. Source of unidentified growth factors. Trademark of Chas. Pfizer & Co., Inc. for purified extraction product from streptomyces fermentation, a Trace mineral mix (%): Mn 6.0, Fe 2.0, Cu 0.20, Co 0.20,1 0.12, Zn 0.006, Ca 27,0. * Vitamin mix (%): Vitamin A (10,000 I.U./gm.) 0.10, vitamin D 3 (3,000 I.C.U./gm.) 0.05, vitamin E (20,000 I.U./gm.) 0.05, riboflavin (4 gm./lb.) 0.06, calcium pantothenate (45% activity) 0.0017, niacin 0.0025, choline chloride (25% dry mix) 0.20, vitamin Bi2 (9mg./lb.) 0.125, menadione sodium bisulfite 0.0003, butylated hydroxy anisole (an antioxidant) 0.0125, DL-methionine 0.14. 6 Oyster shell offered free choice brings estimated Ca level up to 2.3%.
1 2
0.47 0.605
936 21
Productive energy, Cal./lb. % Protein, calculated Inorganic phosphorus, calcul. % Calcium, calculated
100.
— — 0.742
— — 0.742
100.
0.012
0.012
— — — 0.40
1.90
1.60
— — 1.30
33.50
•
60.54
63.45
—
A2
Ai
Broiler expt. 1
Total
Wheat, std. middlings Alfalfa meal, dehyd., 17% prot. Soybean oil meal, solv., 44% prot. Soybean oil meal, solv., 50% prot. Fat, stab, animal Cellulose1 Calcium carbonate Dicalcium phosphate Phosphoric acid, 85% Sodium phosphate, dibasic Disodium carbonate Sodium chloride Magnesium sulfate, 79% Vigofac2 Trace mineral mix3 Vitamin mix4
Corn meal, yellow
Ingredients
TABLE 1.
om http://ps.oxfordjournals.org/ at Fordham University Library on June 22, 2015
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753
PERIODIC REMOVAL OF CALCIUM
TABLE 2.—Influence of dietary calcium level on chick growth, feed utilization and bone ash (experiments 1-4) Weight gain, gm.
Dietary calcium
0-14 days
Expt. N o . and ration
15-28 days 1
%
As% control
Gm.
As% control
Gm.
As% control
Feed utilization 0-28 days 1 Lb. gain/lb. feed
Index of perform-
Bone ash Gain X 28th day, As% feed util- A s % % control ization control
Ai Aj
0.605 1.100
55.0
114 126
90.5
240 257
93.4
0.533 0.548
97.3
189 210
90.0
2
Bi B2 Bs
0.458 0.702 0.930
49.2 75.5
139* 148 « 168
82.7 88.0
280* 311 319
87.8 97.5
0.647 0.652 0.681
95.0 95.7
271 299 332
81.6 90.1
3
Bi B2 Bs
0.458 0.702 0.930
49.2 75.5
89 95 97
91.7 97.9
72 85 80
90.0 106.2
0.431 0.436 0.437
98.6 99.8
69 78 77
89.6 101.3
4
Ci Cj Cs
0.1382 1.138 2.138
12.1
80* 152 142
52.6
256* 302 284*
84.8
0.535 0.540 0.550
99.1
180 245 234
187.9
93.4
94.0
101.8
73.5 95.5
45.6 48.2 47.9
43.8 46.2 46.7
1
For experiment 3, periods were 15-25 days and 0-25 days. Calcium level increased to 0.338% at 14 days. * Significantly different from control, P <0.05.
2
The effect of feeding an extra-high level of calcium approximately double the National Research Council (1954) requirements, was also studied in experiment 4. There was 7% less gain than in controls, which was evident after 14 days on 2.138% calcium, and 6% less growth in the 15-28 day period. The bone ash data of this experiment showed no increase due to the very high calcium and a non-significant reduction (5.2%) due to the low calcium diet. In experiment 2 there was an appreciable, but not significant reduction of bone ash in birds on 0.458% Ca. There was no effect on this value for birds receiving 0.702% Ca. In experiment 5, broiler chicks were maintained on sub-optimal dietary calcium for periods of from two to eight days at various stages of their life cycle. Growth data are summarized in Table 3. Analysis of variance of the weekly weights Feed utilization data (lb. gain/lb. feed) showed that growth was significantly deare presented for the overall 0-28 day pressed for three weeks only when 0.2% period. These show that there was no decalcium was fed from 0 to 8 days of age. pression, or only slight depression, of feed efficiency from the low-calcium rations, Weight gains were somewhat lower than even in the group which started at 0.138% those of controls at succeeding weekly Ca, and changed at two weeks to 0.338% intervals up to 8 weeks, but these differences were not significant. No consequenCa.
calcium was raised to 0.338% by addition of calcium carbonate. Significant retardation of growth on low calcium continued into the 15-28 day period only in the group of experiment 2 receiving 0.458% Ca and the group of experiment 4 initially on 0.138% Ca, later raised to 0.338%. Even under the extreme conditions of experiment 4, it is of interest that, although growth was 47% below that of the control during the first two weeks, it was only 15% below during the second two weeks. In the other low-calcium groups during the second two weeks, there was appreciable, but not significant, retardation of gain as compared with the controls. In all low-calcium groups, the retardation was relatively less during the second two-week period even though calcium levels were unchanged except for the group in experiment 4 initially on only 0.138% Ca.
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1
754
G. A. D O N O V A N , W.
C. SHERMAN, K. E. P R I C E AND J. H .
HARE
TABLE 3.—Influence on weekly chick weights ofreduced dietary calcium levels for short periods at various stages of the growth cycle (experiment 5)
Period of low calcium, days
Dietary calcium1
Avg. male+female weights at days indicated, gm.
Avg. male weights at days indicated, gm.
7
14
21
28
35
42
49
57
Control
1.2
98
193
328
495
656
885
1,128
1,374
0- 2 0- 2
0.2 0.6
104** 95
200 193
332 332
501 495
664 651
895 864
1,141 1,108
1,367 1,312
0- 4 0- 4
0.2 0.6
96 102
193 198
334 340
497 513
651 683
896 902
1,155 1,138
1,361 1,339
0- 8 0- 8
0.2 0.6
90** 100
175** 200
308** 344**
482 517
641 690
832 908
1,072 1,148
1,299 1,377
28-32 28-32
0.2 0.6
490 496
648 661
868 845
1,124 1,083
1,351 1,284
28-36 28-36
0.2 0.6
511 501
682 668
891 863
1,152 1,115
1,378 1,335
49-53 49-53
0.2 0.6
1,125 1,075
1,343 1,323
49-57 49-57
0.2 0.6
1,119 1,152
1,337 1,343
1,122
1,342
Overall Average
98
193
331
500
663
877
• Except during periods indicated, all lots received the basal diet containing 1.2% Ca. ** Significantly greater or less than control (P<0.01). tial differences in weight gain were noted when 0.2% Ca was fed from 0-2, 0-4 days of age or for 4 to 8 day intervals later in the growth cycle. There was no depression of growth from 0.6% Ca fed 0-2, 0-4, or 0-8 days, or for 4 and 8 day intervals later in the cycle. Two instances appearing in the table of growth enhancement on low calcium are inconsistent with other observations and are not readily explainable. Feed utilization data for experiment 5 are shown in Table 4. An analysis of variance showed no significant differences due to any of the treatments in overall feed utilization from day of age to eight weeks. The bone ash d a t a of experiment 5 are presented in Table 5. Analysis of variance showed a significant treatment (days) effect, b u t no calcium level affect. A multiple range test of the averages in Table 5
showed t h a t , during the 50-57 day period, both low levels of dietary calcium significantly reduced the 57-day bone ash as compared to t h a t of controls on a "norm a l " calcium level. No marked influence on 57-day bone ash was evident from calcium deprivation at short intervals earlier in the growth cycle. Laying Birds.—Data of the laying hen trial are presented in Table 6. During the five-day experimental period, egg production dropped 9.6% in hens receiving 0.119% Ca, as compared with a six-week pre-experimental period. There was only 0.25% drop during the experimental period for hens on a normal diet (1.34% Ca plus free-choice oyster shell). Both groups experienced about the same decline postexperimentally in comparison with the pre-experimental period, indicating no carry-over of the effect of drastic lowering of calcium for 5 days.
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%.
PERIODIC REMOVAL or CALCIUM
755
TABLE 4.—Influence on weekly feed conversion of reduced dietary calcium for short periods at various stages of the growth cycle {experiment 5) Period Dietary of low 1 calcium, calcium % days
Av. male+female feed utilization at periods indicated (lb. gain/lb. feed)
Av. male feed utilization at periods indicated (lb. gain/lb. feed)
0-7 days
8-14 days
15-21 days
22-28 days
29-35 days
36-42 days
43-49 days
50-57 days
0-57 days
1.2
.704
.645
.585
.546
.465
.431
.454
.406
.481
0- 2 0- 2
0.2 0.6
.671 .610
.671 .667
.585 .592
.549 .546
.490 .438
.435 .435
.478 .467
.402 .403
.474 .450
0- 4 0- 4
0.2 0.6
.709 .671
.671 .625
.595 .595
.549 .552
.452 .461
.438 .429
.481 .448
.373 .392
.481 .450
0- 8 0- 8
0.2 0.6
.704 .735
.645 .649
.621 .588
.599* .535
.448 .469
.424 .427
.478 .450
.420 .402
.485 .431
28-32 28-32
0.2 0.6
—
—
—
.429 .452
.450 .410
.457 .463
.395 .452
.465 .474
28-36 28-36
0.2 0.6
—
—
—.
.450 .490
.382 .388
.472 .526
.395 .418
.457 .485
49-53 49-53
0.2 0.6
—
—
—
—
.391 .433
.465 .485
49-57 49-57
0.2 0.6
—
—
—
.448 .412
.446 .481
.552
.457
.410
.472
.685
Overall Average
— — — — .654
— — .595
— — •
.424
—
.469
[
Except during periods indicated, all lots received the basal diet containing 1.2% Ca. * Significantly different from control (P<0.05). TABLE 5.^-The influence of reduced dietary calcium levels for short periods on eight-week bone ash Bone ash, % Period of low Ca intake, days 1 0- 2 0- 4 0- 8 29-32 29-36 50-53 50-57 Average
Average
Dietary Ca, 0.2%
Dietary Ca, 0.6%
57.4 56.3 57.2 57.4 57.1 57.1 55.8
56.8 56.3 57.3 56.7 57.1 56.9 56.0
57.1 56.3 57.2 57.1 57.1 57.0 55.9**
56.9
56.8
56.8
Control (1.2% Ca)
TABLE 6.—Influence of short-term low-level dietary calcium on egg production
Diet
Ei E2
56.7
1
Except during periods indicated, all lots received the basal diet containing 1.2% Ca. ** Significantly less than control (P<0.01). CONCLUSIONS
So far as effects on growth, feed efficiency and bone ash are concerned, it would appear practical to omit major calcium
% egg production 1 Calcium %o£ Period 2 Period 3 diet Period Period as a % of Period as a % of 1 2 period 1 period 1 3 0.12 1.340*
62.0 67.1
56.0 67.0
90.3 99.8
56.6 59.2
91.3 88.2
1 Period 1=6 weeks, Period 2=5 days, Period 3 =6 weeks. 2 All groups received Diet E2-r-free choice oyster shell during periods 1 and 3 thus increasing Ca intake to an estimated 2.3%.
sources from the diet of broilers for periods not exceeding 4 days, if started at day of age and not exceeding 8 days if started after 4 weeks of age, in conjunction with use of high-level antibiotics, in order to enhance therapeutic effectiveness. For layers, egg production was lowered during the five days on low calcium, but the depression did not extend beyond the withdrawal period. SUMMARY
The effect on growing chickens of low
Downloaded from http://ps.oxfordjournals.org/ at Fordham University Library on June 22, 2015
Control
756
G. A. D O N O V A N , W.
C. SHERMAN, K. E. P R I C E AND J. H .
REFERENCES Association of Official Agricultural Chemists, 1955. Official Methods of Analysis, 8th Ed. Ewing, W. R., 1951. Poultry Nutrition, 4th Ed. W. R. Ewing, Publisher, S. Pasadena, Calif, pp. 530-531. Donovick, R., A. P. Boyan, P. Canales and F. Pansy, 1948. The influence of certain substances on the activity of streptomycin. III. Differential effects of various electrolytes on the action of streptomycin. J. Bact. 56: 125-137. National Research Council, 1954. Nutrient requirements for domestic animals. No. 1. Nutrient requirements for poultry. Newton, B. A., 1953. Reversal of the antibacterial effects of polymyxin by divalent cations. Nature, 172: 160-161. Price, K. E., M. J. Gallian, C. D. Heather and H. G. Luther, 1955. The influence of milk and other media on antibiotic sensitivity of mastitis organisms. Antibiotics Annual, 1955-56, pp. 753-762. New York Medical Encyclopedia, Inc. Price, K. E., Z. ZoUi, Jr., J. C. Atkinson and A. P. Collins, 1958. Antibiotic inhibitors. III. Reversal of calcium inhibition of intestinal absorption of oxytetracycline in chickens by certain acids and acid salts. Antibiotics Annual 195859, pp. 1020-1032. New York Medical Encyclopedia, Inc. Price, K. E., Z. ZoUi, Jr. and J. H. Hare, 1959a. The effect of dietary calcium-phosphorus and/or supplementation with terephthalic acid upon serum antibiotic levels of chickens. Poultry Sci. 38: 233-235. Price, K. E., and Z. ZoUi, Jr., 1959b. The influence of dietary calcium-phosphorus and terephthalic acid on antibiotic control of experimental infectious synovitis. Avian Diseases, 3: 135-156. Schoenhard, D. E., and H. J. Stafseth, 1953. Some observations on the in vitro action of neomycin. Antib. Chemo. 3:41-49. Weinberg, E. D., 1955. The effect of metallic cations on the anti-streptomyces activity of anhydrochlortetracycline. J. Pharmacol. Exper. Therap. 115: 61-73.
Downloaded from http://ps.oxfordjournals.org/ at Fordham University Library on June 22, 2015
calcium diets was investigated. With dietary calcium levels ranging from 0.702% to 0.138%, half the groups showed significant retardation of growth during the first two weeks, but significant retardation continued in only two groups in the second two weeks. Other low-calcium groups showed appreciable, but not significant retardation of gain over the four weeks. There was no depression or only slight depression of feed utilization measured over four weeks. Bone ash percentages appeared to be affected only in the groups receiving 0.138% a n d " 0 . 4 5 8 % Ca. I n another experiment a calcium level of 0.2% significantly depressed growth only when fed from 0 to 8 days and a significant effect did not carry beyond 3 weeks. No diminution of weight gains was noted when the ration contained only 0.2% Ca from 0-2 or 0-4 days of age or during fourand eight-day intervals at stages from 4 weeks to 8 weeks. There was no depression of growth when only 0.6% Ca was fed 0-2, 0-4, or 0-8 days, for four- and eightday intervals at later stages. Feed utilization was not significantly lowered by either level of restricted calcium in this experiment. Bone ash, determined at 57 days, was lowered only when Ca was reduced to 0.2% or 0.6% in the eight days immediately preceding the determination. With laying hens, egg production was lowered about 1 0 % on 0.119% Ca during a five-day experimental period but this effect did not continue when a normal ration was resumed.
HARE
J . G. OsTRANDER, R . JORDAN, W . J . S T A D E L M A N , J . C. ROGLER AND G. E . VAIL ing results from a research. A report. Purdue University, Mimeo. Reiser, R., 1950. Fatty acid changes in egg yolk of hens on a fat-free and a cottonseed oil ration. J. Nutrition, 40: 429-440. Reiser, R., 1951. The biochemical conversion of conjugated dienoic and trienoic fatty acids. Arch. Biochem. Biophys. 32: 113-120. Riemenschneider, R. W., N. R. Ellis and H. W. Titus, 1938. The fat acids in the lecithin and glyceride fractions of egg yolk. J. Biol. Chem. 126: 255-263. Russell, W. C , M. W. Taylor and H. A. Walker, 1941. The intake and output of fat by the hen on low fat and normal ration. Poultry Sci. 20: 372378. Shorland, F. B., 1951. The fatty acid composition of egg-yolk lipids. New Zealand J. Sci. Technol. B33: 224-229.
The Influence of Periodic Removal of Calcium Sources from Rations of Chickens on Growth and Egg Production G. A. DONOVAN, W. C. SHERMAN, K. E. PRICE 1 AND J. H. HARE Chas. Pfizer & Co., Inc., Terre Haute, Indiana (Received for publication September 18, 1959)
T
HE calcium requirement of young chicks as determined by various workers ranges from 0.66 to 2.14 percent of the diet (Ewing, 1951). This author suggests that the minimum calcium requirement for growing chicks is about 0.70%, but to provide a margin of safety the diet should furnish about 1.0% as recommended by the National Research Council (1954). The value of antibiotics administered in feed or water in the treatment of diseases of poultry has been well documented. Recently it has been demonstrated that the in vitro antibacterial activity of antibiotics is impaired by salts of various metallic ions (Newton, 1953; Weinberg, 1
Present address: Bristol Laboratories, Syracuse,
N.Y.
1955; Donovick el al., 1948; Schoehhard and Stafseth, 1953). The relative susceptibility of 10 antibiotics to inhibition by salts of calcium was determined by Price et al. (1955). They found that streptomycin and neomycin were markedly inhibited, that the tetracycline antibiotics were substantially inhibited, but that the antibacterial action of penicillin and oleandomycin was not influenced by calcium. Apparently part of the same phenomenon is the finding that absorption of oxytetracycline2 from the intestine of the chick is substantially inhibited by calcium ions as shown in experiments with exposed ligated duodenal loops (Price el al., 1958). 2
Inc.
Terramycin, Trademark of Chas. Pfizer & Co.,
Downloaded from http://ps.oxfordjournals.org/ at Fordham University Library on June 22, 2015
depot fat of the fowl as affected by the ingestion of large amounts of different fats. Biochem. J. 28: 96S-977. Dixon, W. J., and F. J. Massey, Jr., 1957. Introduction to Statistical Analysis. New York, McGrawHill Book Company, Inc. Feigenbaum, A. S., and H. Fisher, 1959. The influence of dietary fat on the incorporation of fatty acids into the body and egg fat of the hen. Arch. Biochem. Biophys. 79: 302-306. Fisher, H., and G. A. Leveille, 1957. Observations on the cholesterol, linoleic and linolenic acid content of eggs as influenced by dietary fats. J. Nutrition, 63: 119-129. McCollum, E. V., J. G. Halpin and A. H. Drescher, 1912. Synthesis of lecithin in the hen and the character of the lecithins produced. J. Biol. Chem. 13: 219-224. Miles, S. R., 1957. Drawing conclusions and report-
751
PERIODIC REMOVAL OF CALCIUM
of the life cycle, simulating the periods during which high-level antibiotics might be applied to combat disease. Normal calcium levels were fed during other than the experimental periods. The females in this experiment were removed at 35 days due to lack of space when males were transferred to finishing batteries. Egg Production Study.—One experiment was conducted with S. C. White Leghorn hens eleven months old which had been in production about six months. Rations Ei and E2, Table 1, which provided approximately 0.12% and 1.3% calcium, were tested. There were two floor pen groups of 18 birds on each treatment. RESULTS
Growing Birds.—Results of experiments 1 to 4 on continuous feeding of low calcium from day of age to 28 days are presented in Table 2. The six low-calcium EXPERIMENTAL groups in this series received diets conGrowth Studies.—Five experiments were taining from 0.702% Ca down to 0.138%. conducted with Nichols X Vantress cross- In three of these six groups there was a breeds. In experiments 1 to 4, low calcium significant retardation of gain on low callevels were tested in comparison with cium during the first two weeks (expericalcium levels approximating National ment 2, 0.458% and 0.702% Ca; experiResearch Council (1954) requirements ment 4, 0.138% Ca). There were appreciwhen fed continuously from 0 to 28 days, able, but not significant, reductions in except that, in one trial, observations rate of gain in the other three low-calcium extended only to 25 days because of an groups in the 0-14 day period. That reoutbreak of Newcastle disease. Experi- ductions were not as severe in experiment mental diets are shown in Table 1. Obser- 3, which was essentially parallel to experivations were made of weight gains in the ment 2, is probably explained by the periods 0-14 days and 14-28 days, of generally slower rate of gain of all groups, feed utilization (lb. gain/lb. feed) 0-28 including the control, because of the presdays, and index of performance (gainX ence of Newcastle disease in experiment feed utilization) 0-28 days. Bone ash de- 3. terminations were made by the method of Especially noteworthy is the group in the A.O. A.C. (1955) on the second phalanx experiment 4 which received the extremely of the second digit of the right foot. The low calcium level of 0.138% for two weeks. fifth experiment was conducted to deter- At 13 to 14 days about a third of these mine the physiological effect of maintain- birds began to hobble around on their ing broiler chicks on sub-optimal calcium hocks and were unable to stand and walk for periods of 2 to 8 days at various stages normally so that, at this time, the dietary
Downloaded from http://ps.oxfordjournals.org/ at Fordham University Library on June 22, 2015
This action is reflected in enhancement of blood oxytetracycline levels with decreases of dietary calcium levels or with simultaneous introduction into the gut of compounds which bind calcium (Price et al, 1959a). From this it follows that high-level antibiotic administration via either the feed or drinking water for control of diseases should be more effective in the presence of low dietary calcium levels for short periods of time. Experimental evidence of this has been obtained by Price et al. (1959b). Before dietary calcium alteration is adopted in practice, it is essential to know whether a temporary reduction of the level of this mineral in the feed adversely affects growth or egg production. This question was investigated in the following experiments.
— _
— — _^_ 32.94 — 0.55 — — 1.90 — — — 0.40
0.47 1.100
936 21
•
0.46 0.702
24
0.46 0.458
1,080
24
100.
0.10 0.742
— 0.46 — 0.61 — 0.30
1,080
100.
0.30 0.10 0.742
—
— 0.60
— — 1.28 — 0.46 — 0.61 1.29
10.01
— — — — 38.85
47.04
B2
9.53
— — — — 38.62
48.36
Bi B,
0.46 0.930
24
1,080
100.
0.10 0.742
— 0.46 — 0.61 — 0.30
1.29
— 1.20
10.45
— —. — — 39.07
45.78
Broiler expt. 2 and 3
0.55 0.138
24
1,008
100.
0.742
0.55 1.138
24
1,008
100.
0.742
0.52
— • — 0.10
— 1.60 — 0.44
— — 0.10
—
9.06 2.63 2.63
— — — — 39.60
42.78
C»
0.44 0.52
•
— — 1.60
9.06 5.26
— .— — ~39.60
42.78
Ci
100.
0.742
0.52
— — 0.10
— 5.26 — 1.60 — 0.44
9.06
— — — — 39.60
42.78
a
0.55 2.138
24
1,008
Broiler expt. 4
Composition of rations (%)
0.55 0.20
990 22
100.
—
0.30 0.10 0.742
0.52
— — 0.44 1.60
4.90 2.63 0.18
— — — — 33.52
55.25
Di
0.55 0.60
990 22
100.
—
0.30 0.10 0.742
0.52
— — 0.44 1.60
4.90 1.58 1.23
— — — — 33.52
55.25
D2
Da
0.55 1.20
990 22
100.
—
0.30 0.10 0.742
0.52
— — 1.60 — 0.44
2.81
4.90
— — — — 33.52
55.25
Broiler expt. 5
0.50 0.119
968 17
100.
—
•
0.742
—
0.015
— — — — — 1.76 — 0.45
2.27
63.20 3.00 5.00 2.00 21.56
Ei
0.50 1.340«
949 17
100.
— —
0.742
0.015
— — — 0.45
2.13
—• — 1.83 2.27
61.00 3.00 5.00 2.00 21.56
Es
Layer expt.
Solka-floc. Source of unidentified growth factors. Trademark of Chas. Pfizer & Co., Inc. for purified extraction product from streptomyces fermentation, a Trace mineral mix (%): Mn 6.0, Fe 2.0, Cu 0.20, Co 0.20,1 0.12, Zn 0.006, Ca 27,0. * Vitamin mix (%): Vitamin A (10,000 I.U./gm.) 0.10, vitamin D 3 (3,000 I.C.U./gm.) 0.05, vitamin E (20,000 I.U./gm.) 0.05, riboflavin (4 gm./lb.) 0.06, calcium pantothenate (45% activity) 0.0017, niacin 0.0025, choline chloride (25% dry mix) 0.20, vitamin Bi2 (9mg./lb.) 0.125, menadione sodium bisulfite 0.0003, butylated hydroxy anisole (an antioxidant) 0.0125, DL-methionine 0.14. 6 Oyster shell offered free choice brings estimated Ca level up to 2.3%.
1 2
0.47 0.605
936 21
Productive energy, Cal./lb. % Protein, calculated Inorganic phosphorus, calcul. % Calcium, calculated
100.
— — 0.742
— — 0.742
100.
0.012
0.012
— — — 0.40
1.90
1.60
— — 1.30
33.50
•
60.54
63.45
—
A2
Ai
Broiler expt. 1
Total
Wheat, std. middlings Alfalfa meal, dehyd., 17% prot. Soybean oil meal, solv., 44% prot. Soybean oil meal, solv., 50% prot. Fat, stab, animal Cellulose1 Calcium carbonate Dicalcium phosphate Phosphoric acid, 85% Sodium phosphate, dibasic Disodium carbonate Sodium chloride Magnesium sulfate, 79% Vigofac2 Trace mineral mix3 Vitamin mix4
Corn meal, yellow
Ingredients
TABLE 1.
om http://ps.oxfordjournals.org/ at Fordham University Library on June 22, 2015
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753
PERIODIC REMOVAL OF CALCIUM
TABLE 2.—Influence of dietary calcium level on chick growth, feed utilization and bone ash (experiments 1-4) Weight gain, gm.
Dietary calcium
0-14 days
Expt. N o . and ration
15-28 days 1
%
As% control
Gm.
As% control
Gm.
As% control
Feed utilization 0-28 days 1 Lb. gain/lb. feed
Index of perform-
Bone ash Gain X 28th day, As% feed util- A s % % control ization control
Ai Aj
0.605 1.100
55.0
114 126
90.5
240 257
93.4
0.533 0.548
97.3
189 210
90.0
2
Bi B2 Bs
0.458 0.702 0.930
49.2 75.5
139* 148 « 168
82.7 88.0
280* 311 319
87.8 97.5
0.647 0.652 0.681
95.0 95.7
271 299 332
81.6 90.1
3
Bi B2 Bs
0.458 0.702 0.930
49.2 75.5
89 95 97
91.7 97.9
72 85 80
90.0 106.2
0.431 0.436 0.437
98.6 99.8
69 78 77
89.6 101.3
4
Ci Cj Cs
0.1382 1.138 2.138
12.1
80* 152 142
52.6
256* 302 284*
84.8
0.535 0.540 0.550
99.1
180 245 234
187.9
93.4
94.0
101.8
73.5 95.5
45.6 48.2 47.9
43.8 46.2 46.7
1
For experiment 3, periods were 15-25 days and 0-25 days. Calcium level increased to 0.338% at 14 days. * Significantly different from control, P <0.05.
2
The effect of feeding an extra-high level of calcium approximately double the National Research Council (1954) requirements, was also studied in experiment 4. There was 7% less gain than in controls, which was evident after 14 days on 2.138% calcium, and 6% less growth in the 15-28 day period. The bone ash data of this experiment showed no increase due to the very high calcium and a non-significant reduction (5.2%) due to the low calcium diet. In experiment 2 there was an appreciable, but not significant reduction of bone ash in birds on 0.458% Ca. There was no effect on this value for birds receiving 0.702% Ca. In experiment 5, broiler chicks were maintained on sub-optimal dietary calcium for periods of from two to eight days at various stages of their life cycle. Growth data are summarized in Table 3. Analysis of variance of the weekly weights Feed utilization data (lb. gain/lb. feed) showed that growth was significantly deare presented for the overall 0-28 day pressed for three weeks only when 0.2% period. These show that there was no decalcium was fed from 0 to 8 days of age. pression, or only slight depression, of feed efficiency from the low-calcium rations, Weight gains were somewhat lower than even in the group which started at 0.138% those of controls at succeeding weekly Ca, and changed at two weeks to 0.338% intervals up to 8 weeks, but these differences were not significant. No consequenCa.
calcium was raised to 0.338% by addition of calcium carbonate. Significant retardation of growth on low calcium continued into the 15-28 day period only in the group of experiment 2 receiving 0.458% Ca and the group of experiment 4 initially on 0.138% Ca, later raised to 0.338%. Even under the extreme conditions of experiment 4, it is of interest that, although growth was 47% below that of the control during the first two weeks, it was only 15% below during the second two weeks. In the other low-calcium groups during the second two weeks, there was appreciable, but not significant, retardation of gain as compared with the controls. In all low-calcium groups, the retardation was relatively less during the second two-week period even though calcium levels were unchanged except for the group in experiment 4 initially on only 0.138% Ca.
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1
754
G. A. D O N O V A N , W.
C. SHERMAN, K. E. P R I C E AND J. H .
HARE
TABLE 3.—Influence on weekly chick weights ofreduced dietary calcium levels for short periods at various stages of the growth cycle (experiment 5)
Period of low calcium, days
Dietary calcium1
Avg. male+female weights at days indicated, gm.
Avg. male weights at days indicated, gm.
7
14
21
28
35
42
49
57
Control
1.2
98
193
328
495
656
885
1,128
1,374
0- 2 0- 2
0.2 0.6
104** 95
200 193
332 332
501 495
664 651
895 864
1,141 1,108
1,367 1,312
0- 4 0- 4
0.2 0.6
96 102
193 198
334 340
497 513
651 683
896 902
1,155 1,138
1,361 1,339
0- 8 0- 8
0.2 0.6
90** 100
175** 200
308** 344**
482 517
641 690
832 908
1,072 1,148
1,299 1,377
28-32 28-32
0.2 0.6
490 496
648 661
868 845
1,124 1,083
1,351 1,284
28-36 28-36
0.2 0.6
511 501
682 668
891 863
1,152 1,115
1,378 1,335
49-53 49-53
0.2 0.6
1,125 1,075
1,343 1,323
49-57 49-57
0.2 0.6
1,119 1,152
1,337 1,343
1,122
1,342
Overall Average
98
193
331
500
663
877
• Except during periods indicated, all lots received the basal diet containing 1.2% Ca. ** Significantly greater or less than control (P<0.01). tial differences in weight gain were noted when 0.2% Ca was fed from 0-2, 0-4 days of age or for 4 to 8 day intervals later in the growth cycle. There was no depression of growth from 0.6% Ca fed 0-2, 0-4, or 0-8 days, or for 4 and 8 day intervals later in the cycle. Two instances appearing in the table of growth enhancement on low calcium are inconsistent with other observations and are not readily explainable. Feed utilization data for experiment 5 are shown in Table 4. An analysis of variance showed no significant differences due to any of the treatments in overall feed utilization from day of age to eight weeks. The bone ash d a t a of experiment 5 are presented in Table 5. Analysis of variance showed a significant treatment (days) effect, b u t no calcium level affect. A multiple range test of the averages in Table 5
showed t h a t , during the 50-57 day period, both low levels of dietary calcium significantly reduced the 57-day bone ash as compared to t h a t of controls on a "norm a l " calcium level. No marked influence on 57-day bone ash was evident from calcium deprivation at short intervals earlier in the growth cycle. Laying Birds.—Data of the laying hen trial are presented in Table 6. During the five-day experimental period, egg production dropped 9.6% in hens receiving 0.119% Ca, as compared with a six-week pre-experimental period. There was only 0.25% drop during the experimental period for hens on a normal diet (1.34% Ca plus free-choice oyster shell). Both groups experienced about the same decline postexperimentally in comparison with the pre-experimental period, indicating no carry-over of the effect of drastic lowering of calcium for 5 days.
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%.
PERIODIC REMOVAL or CALCIUM
755
TABLE 4.—Influence on weekly feed conversion of reduced dietary calcium for short periods at various stages of the growth cycle {experiment 5) Period Dietary of low 1 calcium, calcium % days
Av. male+female feed utilization at periods indicated (lb. gain/lb. feed)
Av. male feed utilization at periods indicated (lb. gain/lb. feed)
0-7 days
8-14 days
15-21 days
22-28 days
29-35 days
36-42 days
43-49 days
50-57 days
0-57 days
1.2
.704
.645
.585
.546
.465
.431
.454
.406
.481
0- 2 0- 2
0.2 0.6
.671 .610
.671 .667
.585 .592
.549 .546
.490 .438
.435 .435
.478 .467
.402 .403
.474 .450
0- 4 0- 4
0.2 0.6
.709 .671
.671 .625
.595 .595
.549 .552
.452 .461
.438 .429
.481 .448
.373 .392
.481 .450
0- 8 0- 8
0.2 0.6
.704 .735
.645 .649
.621 .588
.599* .535
.448 .469
.424 .427
.478 .450
.420 .402
.485 .431
28-32 28-32
0.2 0.6
—
—
—
.429 .452
.450 .410
.457 .463
.395 .452
.465 .474
28-36 28-36
0.2 0.6
—
—
—.
.450 .490
.382 .388
.472 .526
.395 .418
.457 .485
49-53 49-53
0.2 0.6
—
—
—
—
.391 .433
.465 .485
49-57 49-57
0.2 0.6
—
—
—
.448 .412
.446 .481
.552
.457
.410
.472
.685
Overall Average
— — — — .654
— — .595
— — •
.424
—
.469
[
Except during periods indicated, all lots received the basal diet containing 1.2% Ca. * Significantly different from control (P<0.05). TABLE 5.^-The influence of reduced dietary calcium levels for short periods on eight-week bone ash Bone ash, % Period of low Ca intake, days 1 0- 2 0- 4 0- 8 29-32 29-36 50-53 50-57 Average
Average
Dietary Ca, 0.2%
Dietary Ca, 0.6%
57.4 56.3 57.2 57.4 57.1 57.1 55.8
56.8 56.3 57.3 56.7 57.1 56.9 56.0
57.1 56.3 57.2 57.1 57.1 57.0 55.9**
56.9
56.8
56.8
Control (1.2% Ca)
TABLE 6.—Influence of short-term low-level dietary calcium on egg production
Diet
Ei E2
56.7
1
Except during periods indicated, all lots received the basal diet containing 1.2% Ca. ** Significantly less than control (P<0.01). CONCLUSIONS
So far as effects on growth, feed efficiency and bone ash are concerned, it would appear practical to omit major calcium
% egg production 1 Calcium %o£ Period 2 Period 3 diet Period Period as a % of Period as a % of 1 2 period 1 period 1 3 0.12 1.340*
62.0 67.1
56.0 67.0
90.3 99.8
56.6 59.2
91.3 88.2
1 Period 1=6 weeks, Period 2=5 days, Period 3 =6 weeks. 2 All groups received Diet E2-r-free choice oyster shell during periods 1 and 3 thus increasing Ca intake to an estimated 2.3%.
sources from the diet of broilers for periods not exceeding 4 days, if started at day of age and not exceeding 8 days if started after 4 weeks of age, in conjunction with use of high-level antibiotics, in order to enhance therapeutic effectiveness. For layers, egg production was lowered during the five days on low calcium, but the depression did not extend beyond the withdrawal period. SUMMARY
The effect on growing chickens of low
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Control
756
G. A. D O N O V A N , W.
C. SHERMAN, K. E. P R I C E AND J. H .
REFERENCES Association of Official Agricultural Chemists, 1955. Official Methods of Analysis, 8th Ed. Ewing, W. R., 1951. Poultry Nutrition, 4th Ed. W. R. Ewing, Publisher, S. Pasadena, Calif, pp. 530-531. Donovick, R., A. P. Boyan, P. Canales and F. Pansy, 1948. The influence of certain substances on the activity of streptomycin. III. Differential effects of various electrolytes on the action of streptomycin. J. Bact. 56: 125-137. National Research Council, 1954. Nutrient requirements for domestic animals. No. 1. Nutrient requirements for poultry. Newton, B. A., 1953. Reversal of the antibacterial effects of polymyxin by divalent cations. Nature, 172: 160-161. Price, K. E., M. J. Gallian, C. D. Heather and H. G. Luther, 1955. The influence of milk and other media on antibiotic sensitivity of mastitis organisms. Antibiotics Annual, 1955-56, pp. 753-762. New York Medical Encyclopedia, Inc. Price, K. E., Z. ZoUi, Jr., J. C. Atkinson and A. P. Collins, 1958. Antibiotic inhibitors. III. Reversal of calcium inhibition of intestinal absorption of oxytetracycline in chickens by certain acids and acid salts. Antibiotics Annual 195859, pp. 1020-1032. New York Medical Encyclopedia, Inc. Price, K. E., Z. ZoUi, Jr. and J. H. Hare, 1959a. The effect of dietary calcium-phosphorus and/or supplementation with terephthalic acid upon serum antibiotic levels of chickens. Poultry Sci. 38: 233-235. Price, K. E., and Z. ZoUi, Jr., 1959b. The influence of dietary calcium-phosphorus and terephthalic acid on antibiotic control of experimental infectious synovitis. Avian Diseases, 3: 135-156. Schoenhard, D. E., and H. J. Stafseth, 1953. Some observations on the in vitro action of neomycin. Antib. Chemo. 3:41-49. Weinberg, E. D., 1955. The effect of metallic cations on the anti-streptomyces activity of anhydrochlortetracycline. J. Pharmacol. Exper. Therap. 115: 61-73.
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calcium diets was investigated. With dietary calcium levels ranging from 0.702% to 0.138%, half the groups showed significant retardation of growth during the first two weeks, but significant retardation continued in only two groups in the second two weeks. Other low-calcium groups showed appreciable, but not significant retardation of gain over the four weeks. There was no depression or only slight depression of feed utilization measured over four weeks. Bone ash percentages appeared to be affected only in the groups receiving 0.138% a n d " 0 . 4 5 8 % Ca. I n another experiment a calcium level of 0.2% significantly depressed growth only when fed from 0 to 8 days and a significant effect did not carry beyond 3 weeks. No diminution of weight gains was noted when the ration contained only 0.2% Ca from 0-2 or 0-4 days of age or during fourand eight-day intervals at stages from 4 weeks to 8 weeks. There was no depression of growth when only 0.6% Ca was fed 0-2, 0-4, or 0-8 days, for four- and eightday intervals at later stages. Feed utilization was not significantly lowered by either level of restricted calcium in this experiment. Bone ash, determined at 57 days, was lowered only when Ca was reduced to 0.2% or 0.6% in the eight days immediately preceding the determination. With laying hens, egg production was lowered about 1 0 % on 0.119% Ca during a five-day experimental period but this effect did not continue when a normal ration was resumed.
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