- Email: [email protected]
The Availability of Phosphorus from Single and Combined Phosphates to Chicks
TRANQUILIZERS AND EGG PRODUCTION
lives in broiler production. Poultry Sci. 38: 14741475. Garren, H. W., and C. H. Hill, 1957. The effect of continually feeding tranquilizing agents to young White Leghorns. Poultry Sci. 36: 1386-1387. Gilbreath, J. C , L. F. Garvin and Q. B. Welch, 1959. Effect of orally administered reserpine on egg production and quality. Poultry Sci. 38: 535538. Snedecor, G. W., 1956. Statistical Methods, 5th ed. The Iowa State College Press, Ames, Iowa. Van Matre, N. S., R. E. Burger and F. W. Lorenz, 1957. Resistance to heat stress following administration of tranquilizing drugs. Poultry Sci. 36: 1165. Weiss, H. S., 1959. The protective effect of serpasil in laying birds against high lethal temperature. Rutgers Conference on Serpasil: 51-57. Zlatkis, A. B., B. Zak and A. J. Boyle, 1953. A new method for the direct determination of serum cholesterol. J. Lab. Clin. Med. 41:486.
The Availability of Phosphorus from Single and Combined Phosphates to Chicks T. S. NELSON AND H. T. PEELER International Minerals and, Chemical Corporation, Skokie, Illinois (Received for publication November 30. 1960)
prior to 1955 on the utilizaSTTJDIES tion of the phosphorus in various chemical and commercial grade phosphates by the chick have been reviewed by Motzok et al. (1956). In most of the work reviewed no comparative values of the various phosphates were given. Gillis et al. (1954) developed a biological assay whereby the comparative availability of phosphorus contained in various phosphates could be measured. Differences in the quality of phosphates could readily be shown when this technique was used. However, some sources, such as soft phosphate, could not be assayed by this method because of their low availabilities. A practical diet was used to ascertain the available phosphorus in these supplements.
Wilcox et al. (1954, 1955) found that soft phosphate was poorly utilized by turkey poults. The combination of dibasic calcium phosphate, U.S.P., and soft phosphate was utilized to the extent of available phosphorus contributed by each supplement when percentage bone ash was the criterion. Motzok et al. (1956) showed a significant depression in the growth of chicks when soft phosphate replaced a portion or all of the monobasic calcium phosphate. In contrast to these results, Baruah et al. (1960a) reported that the phosphorus in soft phosphate was highly available to chicks. The biological assay such as that described by Gillis et al. (1954) gives a critical measure of phosphate availability
Downloaded from http://ps.oxfordjournals.org/ at Kainan University on April 25, 2015
Association of Official Agricultural Chemists, 1955. Official and Tentative Methods of Analysis, 8th ed., Washington, D. C. Babcock, M. J., and M. W. Taylor, 1957. Effect of meprobamate on growth and feed efficiency of chickens. Poultry Sci. 36: 485-487. Burger, R. E., and F. W. Lorenz, 1960. Pharmacologically induced resistance to heat shock. Poultry Sci. 39: 477-482. Burger, R. E., N. S. Van Matre and F. W. Lorenz, 1959. Growth and mortality of chicks and poults fed tranquilizing drugs. Poultry Sci. 38: 508-512. Carlson, C. W., 1959a. The growth response of chicks to reserpine as affected by breed and energy-protein levels. Poultry Sci. 38: 1194. Carlson, C. W., 1959b. Control of a field outbreak of dissecting aneurysms and laying hen studies with reserpine. Rutgers Conference on Serpasil: 29-35. Duncan, D. B., 1955. Multiple range and multiple F tests. Biometrics, 11: 1-42. Fritz, J. C , F. D. Wharton, Jr. and L. J. Classen, 1959. Experiments with tranquilizers and seda-
1321
1322
T. S. NELSON AND H. T. PEELER
EXPERIMENTAL
The basal diets used in these experiments are given in Tables 1 and 2. The phosphorus content of the purified diet was too low to sustain chick life beyond 10 days. The practical diet contained sufficient phosphorus to sustain the chicks during the experimental period. The calcium and phosphorus supplements replaced cellulose in order to maintain diets within each experiment isonitrogenous and isocaloric. The calcium-phosphorus ratio was 2:1 in all diets except the purified diets containing soft phosphate, Curacao phosphate or a combination of these two sources. The wide calcium-phosphorus ratios of these supplements made a 2:1 dietary ratio impossible. Calcium carbonate, analytical reagent grade, was the calcium supplement used to balance the ratios in the other diets. In experiments 1 and 2, S. C. White Leghorn day-old chicks were used. Each lot contained 10 male chicks and the experimental period was three weeks. In experiment 3 each lot contained 10 day-old
TABLE 1.—Composition of basal diet, experiments 1 and 2 Ingredients Corn starch Dried blood fibrin Gelatin Liver fraction L Hydrogenated fat (Primex) Vitamin mixture21 Mineral mixture Cellulose and supplements Protein, % Metabolizable energy—Cal./lb. ME/P Phosphorus, % analyzed Calcium, % analyzed
% 57.13 21.50 4.00 1.00 3.00 1.22 2.19 10.00 23.5 1,465 62/1 0.07 0.15
1 Furnished the following amounts per 100 gms. of diet: 0.025 mg. biotin, 0.45 mg. folic acid, 0.50 mg. pyridoxine-HCl, 0.25 mg. menadione sodium bisulfite, 1.1 mg. riboflavin, 1.1 mg. thiamine-HC1, 4.30 mg. Ca pantothenate, 3 fig- vitamin Bi 2 , 5.5 mg. niacin, 25 mg. inositol, 500 I.U. vitamin A, 75 I.C.U. vitamin D3, 5 mg. a-tocopheryl acetate and 160 mg. choline chloride. 2 Furnished the following amounts per 100 gms. of diet: 1.00 gm. KC1, 0.75 gm. iodized salt, 0.35 gm. MgS0 4 , 36 mg. MnS0 4 -H 2 0, 36 mg. FeSCv 7H 2 0, 19.2 mg. ZnC0 3 , 2 mg. CuS0 4 -5H 2 0, 0.34 mg. NaMo0 4 -2H 2 0, 0.3 mg. CoCl 2 -6H 2 0 and 0.28 mg. KI.
TABLE 2.—Composition of basal diet, experiment 3 Ingredients Ground yellow corn Soybean oil meal (50% protein) Hydrogenated fat (Primex) Corn gluten meal Cond. fish solubles Alfalfa meal (17% protein, dehydrated) Iodized salt DL-methionine Vitamins1 MnSCvH 2 0, AR grade ZnC0 3 , AR grade Cellulose and supplements Protein, % Metabolizable energy, Cal./lb. ME/P Phosphorus, % analyzed Calcium, % analyzed
% 55.60 28.50 3.00 3.00 2.00 1.50 0.25 0.075 0.05 0.02 0.0125 6.00 21.8 1,360 62/1 0.37 0.17
1 Furnished the following per 100 gms. of diet: 0.275 mg. riboflavin, 0.33 mg. menadione sodium bisulfite, 0.44 mg. Ca pantothenate, 0.66 mg. niacin, 55 mg. choline chloride, 0.88 jug- vitamin B12, 220 I.U. vitamin A, 50 I.C.U. vitamin D 3 and 0.4 mg. a-tocopheryl acetate.
Downloaded from http://ps.oxfordjournals.org/ at Kainan University on April 25, 2015
in contrast to the "replacement" technique. Day and Hill (1959) reported that soft phosphate could replace 25 to 50% of supplementary defluorinated phosphate in the diets of turkeys to 20 weeks of age. Baruah et al. (1960b) using the "replacement" technique concluded that soft phosphate could replace a portion of the dicalcium phosphate or defluorinated phosphate in chick diets. Because the comparative availabilities of combinations of phosphates have not been determined, it was desirable to ascertain to what extent the chick could utilize these combinations. It was also desirable to know if biological values of phosphates determined with a purified diet could be applied to practical diets.
1323
AVAILABILITY OF PHOSPHORUS
The procedure for determining the availability of experimental phosphates using a standard curve has been reported by Gillis et al. (1954). These workers also described the method for preparation of the bones for ashing. RESULTS AND DISCUSSION
Experiment 1 was conducted to determine the bioligical availability of Curacao phosphate and soft phosphate when fed alone and in combination. I t was necessary to add 0.15% phosphorus from betatricalcium phosphate to the diets in which soft phosphate was tested in order to obtain good livability. Previous results (Table 3) obtained in this laboratory showed that an insufficient number of chicks survived the three-week experimental period when soft phosphate was the only source of supplemental phosphorus. Good livability is necessary to get a valid measure of the biological avail-
TABI.E 3. —Evidence of the low availability of phosphorus in soft phosphate cSupplement i „ i
Phosphorus fc
No. of
.Befa-tricalcium phosphate
0.20 0.25 0.30 0.35
0 0 0 0
Soft phosphate
0.25 0.30 0.35 0.40
90 91 82 60
samples
Av. mor-
taUty
%
ability of the phosphorus in any phosphate source. The results of experiment 1 are given in Table 4. The comparative availability of the phosphorus in Curacao phosphate was 6 8 % . These results agreed with those of Scott et al. (1956). These investigators showed t h a t poults could utilize commercial dicalcium phosphate more efficiently than Curacao phosphate. Curacao phosphate was utilized approximately as well as reagent grade dicalcium phosphate which has been shown to be 7 0 % available to chicks when compared to a betatricalcium phosphate standard. The biological value of soft phosphate was 36. The effect of 0.15% phosphorus supplied by ieto-tricalcium phosphate was eliminated from the calculation by subtracting it from the total phosphorus obtained with the standard curve. The availability of the phosphorus in a mixture containing equal amounts of phosphorus from Curacao phosphate and soft phosphate was 4 2 % . The average biological value of the individual phosphates was 52. The results of this experiment showed t h a t the availability of a combination of phosphates was no better than the average of their individual availabilities. The second experiment was conducted to determine if the results obtained in the
Downloaded from http://ps.oxfordjournals.org/ at Kainan University on April 25, 2015
Peterson X C o b b s White Plymouth Rock male chicks. The experimental period was 4 weeks. All chicks were reared in pens with raised wire mesh floors. H e a t was supplied by electrically heated brooders. Feed and distilled water were supplied ad libitum. The chicks were weighed weekly and feed consumption records were maintained. Percentage bone ash was the criterion used to determine the biological availability of the various phosphates by the chicks. The reference phosphate was betatricalcium phosphate. The standard curve was the regression line obtained by plotting the percentage bone ash obtained at the various levels of added phosphorus against the logarithm of the added phosphorus. The regression line was calculated using the method described by Hegsted (1948) and Almquist (1953).
1324
T. S. N E L S O N AND H. T.
PEELER
TABLE 4. -Biological availability of single and combined phosphates, experiment 1 Biological value
0.20 0.25 0.30 0.35
140 160 166 170
35.57 41.25 43.43 46.16
100
Curacao phosphate (CP) 1
0.30 0.35
144 167
35.55 42.59
68
Soft phosphate (SP)2
0.45 1 0.552
142 159
41.83 41.57
36
CP+SP3
0.60 0.80
148 160
42.35 42.85
42
Be/a-tricalcium phosphate
Supplemental phosphorus
%
Actual
Calculated
%
%
52
1 This diet contained 0.15% phosphorus from Seta-tricalcium phosphate and 0.30% phosphorus from soft phosphate. 2 This diet contained 0.15% phosphorus from Seta-tricalcium phosphate and 0.40% phosphorus from soft phosphate. 3 Each supplement provided 50% of the added phosphorus.
first experiment were reproducible. Different samples of Curacao phosphate and soft phosphate were used in this experiment. In addition, dicalcium phosphate was also tested. Again, as in the first experiment, the diets used to determine the biological value of soft phosphate contained 0.15% phosphorus from beta-tricalcium phosphate. The results of this experiment are given in Table 5. The biological value of dicalcium phosphate was 97 and the values obtained for Curacao phosphate and soft phosphate were in close agreement with those found in the first experiment. The available phosphorus in combinations of these phosphates, with each supplement supplying an equal amount of phosphorus, was very close to the calculated value. The calculated biological values of these phosphate mixtures were made using the averages of the individual phosphates. This experiment confirmed the findings in the first experiment. Poor quality phosphates were not improved when fed as mixtures with higher quality phosphates. The third experiment was conducted to
determine if the comparisons of biological availability obtained when the purified diet was fed could be applied to a test using a practical type diet. The experimental plan and results are shown in Table 6. The biological values of the individual phosphates agreed very closely with those obtained in experiment 2. Thus, the biological values of phosphates obtained with the purified diet can be applied to practical diets. Combinations of these phosphates, when each supplement provided the same proportion of phosphorus, had biological values approximately equal to the averages of the biological values of the supplements tested. Therefore, no synergistic effect of phosphates was observed in this experiment. In order to obtain valid comparisons of phosphates, it is necessary t h a t (1) a sensitive criterion of measure be used, (2) a critically phosphorus deficient diet be fed, (3) the levels of added phosphorus not be in excess of the animal's requirement, and (4) a suitable standard be used for comparison to the phosphates tested. No phosphate, regardless of how good or
Downloaded from http://ps.oxfordjournals.org/ at Kainan University on April 25, 2015
Bone ash
%
Av. wt. gain, gm.
Supplement.
1325
AVAILABILITY OF P H O S P H O R U S
TABLE 5.—Biological availability of single and combined phosphates, experiment Z Biological value
Bone ash
%
Av. wt. gain, gm.
0.20 0.25 0.30 0.35
134 142 169 159
29.71 34.88 39.73 41.76
100
Dicalcium phosphate (DCP)
0.25 0.35
142 168
33.98 41.34
97
Curacao phosphate (CP)
0.30 0.40
144 157
32.63 33.91
68
Soft phosphate (SP)
0.45i 0.55 2
133 131
34.66 36.60
31
DCP+CP3
0.30 0.40
150 156
33.90 38.41
77
82
DCP+SP3
0.40 0.60
133 156
35.73 46.11
68
64
CP+SP3
0.60 0.80
139 138
36.64 38.98
42
49
DCP+CP+SP"
0.45 0.60
142 154
38.41 42.72
63
65
Supplement
Seto-tricalcium
phosphate
Supplemental phosphorus
%
Actual
%
Calculated
%
how poor it may be, can be evaluated when tested in diets containing excess phosphorus. D a y and Hill (1959) fed diets to turkeys which, in terms of biologically available phosphorus, contained phosphorus in excess of the recommendations by the National Research Council (1954) and b y Scott (1960). The available phosphorus contents of the diets containing soft phosphate were calculated assuming this supplement to be only 2 5 % available. When 5 0 % of the phosphorus from defluorinated rock phosphate was replaced by the phosphorus in soft phosphate, the available phosphorus contents of these diets were still in excess of the turkey's requirements for this mineral, based on the National Research Council
recommendations. Based on the recommendations b y Scott (1960) the starter diet was borderline deficient in phosphorus and the remaining diets contained an excess of this mineral. T h e phosphorus replaced in these diets was merely excess phosphorus. No true evaluation of soft phosphate was actually determined. Baruah et al. (1960a) calculated the availability of a defluorinated phosphate and soft phosphate by dividing the percentage bone ash obtained with the test phosphate by the percentage bone ash obtained with the standard phosphate. The respective availabilities of these phosphates were reported as 95.5 and 85.7%. According to this method, the availability of no added phosphorus in the basal diet
Downloaded from http://ps.oxfordjournals.org/ at Kainan University on April 25, 2015
1 This diet contained 0.15% phosphorus from fteto-tricalcium phosphate and 0.30% phosphorus from soft phosphate. 2 This diet contained 0.15% phosphorus from ftcto-tricalcium phosphate and 0.40% phosphorus from soft phosphate. 3 Each supplement provided 50% of the added phosphorus. 4 Each supplement provided 33.3% of the added phosphorus.
1326
T. S. NELSON AND H. T. PEELER TABLE 6.—Biological availability of single and combined, phosphates, experiment 3 Phosphorus
Supplement
Biologi cal value
Av. wt. gain, gm.
Bone ash
Perosis
%
%
Actual Calculated
Added,
Available
%
%
0.04 0.08 0.12 0.16 0.20
0.04 0.08 0.12 0.16 0.20
281 313 305 332 357
32.20 35.69 40.73 42.80 44.13
50 10 0 0 0
100
DCP 2
0.05 0.15
0.05 0.15
301 336
33.56 41.62
44 0
99
CP 3
0.10 0.15
0.07 0.10
295 345
36.11 38.87
33 20
69
SP"
0.20 0.30
0.07 0.11
298 328
35.77 40.06
67 10
36
DCP+CP5
0.10 0.20
0.08 0.17
331 352
36.01 43.60
20 10
83
84
DCP+SP6
0.10 0.20
0.07 0.15
317 352
36.56 41.92
30 10
74
68
CP+SP5
0.10 0.20
0.06 0.11
324 310
33.94 40.14
30 40
57
53
DCP+CP+SPS
0.15 0.225
0.10 0.16
323 354
38.20 42.78
33 30
69
68
B-TCP 1
3 4 6 6
%
.Beta-tricalcium phosphate. Dicalcium phosphate. Curacao phosphate. Soft phosphate. Each supplement provided 50% of the added phosphorus. Each supplement provided 33.3% of the added phosphorus.
was 79.9%. The critical, measurable range was actually 79.9-100%. The increases in bone ash attributed to soft phosphate and defluorinated phosphate were then 5.8/20.1 and 15.6/20.1, respectively. The availabilities of these phosphates, based on a scale of 0-100% would be 29 and 78% respectively. Baruah et al. (1960b) in their studies on combinations of soft phosphate with other phosphates fed a basal diet which was not critically deficient in phosphorus nor was a sensitive criterion used. Moreover, the diet used contained a low level of energy, thereby decreasing the range of response to supplemental phosphorus. The requirement of available phosphorus, to produce
maximum bone ash, for chicks fed this diet would be approximately 0.36% based on the results of Peeler et al. (1960). Since the chick reaches maximum weight before attaining maximum bone ash, the requirement for growth on this diet would be less than 0.36% available phosphorus. Therefore, these phosphates, and combinations of them, were not tested over a critical range. The total phosphorus content of poultry rations is meaningless because the chick can utilize phytin phosphorus only to a limited degree (Gillis et al., 1957). The question of whether to express the usable phosphorus in such rations as available phosphorus or inorganic phosphorus oc-
Downloaded from http://ps.oxfordjournals.org/ at Kainan University on April 25, 2015
1 2
%
1327
AVAILABILITY OF PHOSPHORUS
44
/?-Tricalciam Phosphate Dicalcium Phosphate Curacao Phosphate Soft Phosphate
42
I+ff i+m E+m
40
38 x z o o
36
34
"
32 0-01
0O2
0-04
O06
LOGARITHM OF ADDED AVALIABLE
0 0 8 0-10
0-20
PH0SPH0RUS,%
FIG. 1. The relationship of biologically available phosphorus to percentage bone ash.
casionally occurs. The inaccuracy of the term "inorganic phosphate" is clearly demonstrated by the results given in Tables 3, 4 and 5. The entire phosphorus contents of these diets used in these experiments were inorganic. Varying results are obtained at the different levels of the phosphates, depending on the quality of the individual phosphates. However, when these phosphates are expressed on the basis of biologically available phosphorus they contained, the results agreed more closely. The advantage of expressing phosphorus as available phosphorus instead of inorganic phosphorus is also shown in Table 6. Varying results were obtained at specific levels of inorganic phosphorus. However, when expressed in terms of available phosphorus, excellent correlation existed between available phosphorus added and percentage bone ash. This is demonstrated in Figure 1.
The advantage of using percentage bone ash as the criterion of response instead of gain is also demonstrated. The percentage perosis observed in this experiment decreased as the percentage bone ash increased indicating that strong bones are needed to prevent this condition. The highest incidence of perosis was observed in the chicks fed diets containing soft phosphate. SUMMARY
Experiments were conducted with chicks fed purified and practical type diets to determine the biological availability of single and combined phosphate sources. When determined with a purified diet using 6eto-tricalcium phosphate as the reference material, the biological value of one sample of dicalcium phosphate was 97%. The average biological values of two samples each of Curacao phosphate and
Downloaded from http://ps.oxfordjournals.org/ at Kainan University on April 25, 2015
< UJ
1328
T . S. N E L S O N AND H . T. P E E L E R
soft phosphate were 68 a n d 3 4 % respectively.
The
biological
values
of
these
phosphates determined with a practical diet were almost identical with those obtained with the purified diet. Therefore, the results obtained with a purified diet can be applied to practical type diets. When either diet was fed, the biological value of combinations of phosphates was no better t h a n the sum of the available phosphorus contributed b y each source.
Available phosphorus is a better expression
of
the
phosphorus
content
of
poultry rations t h a n either total or inorganic phosphorus. REFERENCES Almquist, H. J., 1953. Evaluation of vitamin requirement data. Poultry Sci. 32: 122-128. Baruah, J. N., R. E. Davis, B. L. Reid and J. R. Couch, 1960a. Phosphorus availability from the ash of unidentified factor sources. Poultry Sci. 39: 840-842. Baruah, J. N., R. E. Davis, B. L. Reid and J. R. Couch, 1960b. Utilization of phosphorus from defluorinated and colloidal phosphate by chicks and laying hens. Poultry Sci. 39: 843-849. Day, E. J., and J. E. Hill, 1959. Zinc, potassium and certain other minerals as additives to practical
Studies with Corn-Soya Laying Diets 4. ENVIRONMENTAL TEMPERATURE—A FACTOR AFFECTING PERFORMANCE OF PULLETS FED DIETS SUBOPTIMAL IN PROTEIN D . J. BRAY AND J. A. G E S E L L
Department of Animal Science, University of Illinois, Urbana, Illinois (Received for publication December 5, 1960)
I
N
OUR
experience,
egg
production
trends of pullets fed diets suboptimal in
peratures was possible. No such effect was observed
when
the
diets
contained
a
protein have varied with season when the
plethora of protein. Conceivably, with a
birds were confined to a building where
plethora of protein in the diet, a reduction
only limited control over prevailing tem-
in the v o l u n t a r y consumption of feed does
Downloaded from http://ps.oxfordjournals.org/ at Kainan University on April 25, 2015
No synergistic effect of such phosphate combinations occurred.
turkey starter and grower rations. Poultry Sci. 38: 646-649. Gillis, M. B., L. C. Norris and G. F. Heuser, 1954. Studies on the biological value of inorganic phosphates. J. Nutrition, 52: 115-126. Gillis, M. B., K. W. Keane and R. A. Collins, 195V. Comparative metabolism of phytate and inorganic P 32 by chicks and poults. J. Nutrition, 62: 13-26. Hegsted, D. M., 1948. The determination of minimum vitamin requirements for growth. J. Nutrition, 35: 399-409. Motzok, I., D. Arthur and H. D. Branion, 1956. Utilization of phosphorus from various phosphate supplements by chicks. Poultry Sci. 35: 627-649. National Research Council, 1954. Nutrient requirements for domestic animals. I. Nutrient requirements for poultry. Peeler, H. T., T. S. Nelson and N. Storer, 1960. The effect of energy on the phosphorus requirement of the chick. Poultry Sci. 39: 1282. Scott, M. L., H. E. Butters and G. O. Ranet, 1956. Studies on bone formation in turkey poults. Proc. Cornell Nutr. Conf. p. 102-109. Scott, M. L., 1960. Turkey rations. New York State College of Agriculture Extension Stencil No. 208. Wilcox, R. A., C. W. Carlson, W. Kohlmeyer and G. G. Gas tier, 1954. The availability of phosphorus from different sources for poults fed purified diets. Poultry Sci. 33: 1010-1014. Wilcox, R. A., C. W. Carlson, W. Kohlmeyer and G. G. Gastler 1955. The availability of phosphorus from different sources for poults fed practical-type diets. Poultry Sci. 34: 1017-1023.
lives in broiler production. Poultry Sci. 38: 14741475. Garren, H. W., and C. H. Hill, 1957. The effect of continually feeding tranquilizing agents to young White Leghorns. Poultry Sci. 36: 1386-1387. Gilbreath, J. C , L. F. Garvin and Q. B. Welch, 1959. Effect of orally administered reserpine on egg production and quality. Poultry Sci. 38: 535538. Snedecor, G. W., 1956. Statistical Methods, 5th ed. The Iowa State College Press, Ames, Iowa. Van Matre, N. S., R. E. Burger and F. W. Lorenz, 1957. Resistance to heat stress following administration of tranquilizing drugs. Poultry Sci. 36: 1165. Weiss, H. S., 1959. The protective effect of serpasil in laying birds against high lethal temperature. Rutgers Conference on Serpasil: 51-57. Zlatkis, A. B., B. Zak and A. J. Boyle, 1953. A new method for the direct determination of serum cholesterol. J. Lab. Clin. Med. 41:486.
The Availability of Phosphorus from Single and Combined Phosphates to Chicks T. S. NELSON AND H. T. PEELER International Minerals and, Chemical Corporation, Skokie, Illinois (Received for publication November 30. 1960)
prior to 1955 on the utilizaSTTJDIES tion of the phosphorus in various chemical and commercial grade phosphates by the chick have been reviewed by Motzok et al. (1956). In most of the work reviewed no comparative values of the various phosphates were given. Gillis et al. (1954) developed a biological assay whereby the comparative availability of phosphorus contained in various phosphates could be measured. Differences in the quality of phosphates could readily be shown when this technique was used. However, some sources, such as soft phosphate, could not be assayed by this method because of their low availabilities. A practical diet was used to ascertain the available phosphorus in these supplements.
Wilcox et al. (1954, 1955) found that soft phosphate was poorly utilized by turkey poults. The combination of dibasic calcium phosphate, U.S.P., and soft phosphate was utilized to the extent of available phosphorus contributed by each supplement when percentage bone ash was the criterion. Motzok et al. (1956) showed a significant depression in the growth of chicks when soft phosphate replaced a portion or all of the monobasic calcium phosphate. In contrast to these results, Baruah et al. (1960a) reported that the phosphorus in soft phosphate was highly available to chicks. The biological assay such as that described by Gillis et al. (1954) gives a critical measure of phosphate availability
Downloaded from http://ps.oxfordjournals.org/ at Kainan University on April 25, 2015
Association of Official Agricultural Chemists, 1955. Official and Tentative Methods of Analysis, 8th ed., Washington, D. C. Babcock, M. J., and M. W. Taylor, 1957. Effect of meprobamate on growth and feed efficiency of chickens. Poultry Sci. 36: 485-487. Burger, R. E., and F. W. Lorenz, 1960. Pharmacologically induced resistance to heat shock. Poultry Sci. 39: 477-482. Burger, R. E., N. S. Van Matre and F. W. Lorenz, 1959. Growth and mortality of chicks and poults fed tranquilizing drugs. Poultry Sci. 38: 508-512. Carlson, C. W., 1959a. The growth response of chicks to reserpine as affected by breed and energy-protein levels. Poultry Sci. 38: 1194. Carlson, C. W., 1959b. Control of a field outbreak of dissecting aneurysms and laying hen studies with reserpine. Rutgers Conference on Serpasil: 29-35. Duncan, D. B., 1955. Multiple range and multiple F tests. Biometrics, 11: 1-42. Fritz, J. C , F. D. Wharton, Jr. and L. J. Classen, 1959. Experiments with tranquilizers and seda-
1321
1322
T. S. NELSON AND H. T. PEELER
EXPERIMENTAL
The basal diets used in these experiments are given in Tables 1 and 2. The phosphorus content of the purified diet was too low to sustain chick life beyond 10 days. The practical diet contained sufficient phosphorus to sustain the chicks during the experimental period. The calcium and phosphorus supplements replaced cellulose in order to maintain diets within each experiment isonitrogenous and isocaloric. The calcium-phosphorus ratio was 2:1 in all diets except the purified diets containing soft phosphate, Curacao phosphate or a combination of these two sources. The wide calcium-phosphorus ratios of these supplements made a 2:1 dietary ratio impossible. Calcium carbonate, analytical reagent grade, was the calcium supplement used to balance the ratios in the other diets. In experiments 1 and 2, S. C. White Leghorn day-old chicks were used. Each lot contained 10 male chicks and the experimental period was three weeks. In experiment 3 each lot contained 10 day-old
TABLE 1.—Composition of basal diet, experiments 1 and 2 Ingredients Corn starch Dried blood fibrin Gelatin Liver fraction L Hydrogenated fat (Primex) Vitamin mixture21 Mineral mixture Cellulose and supplements Protein, % Metabolizable energy—Cal./lb. ME/P Phosphorus, % analyzed Calcium, % analyzed
% 57.13 21.50 4.00 1.00 3.00 1.22 2.19 10.00 23.5 1,465 62/1 0.07 0.15
1 Furnished the following amounts per 100 gms. of diet: 0.025 mg. biotin, 0.45 mg. folic acid, 0.50 mg. pyridoxine-HCl, 0.25 mg. menadione sodium bisulfite, 1.1 mg. riboflavin, 1.1 mg. thiamine-HC1, 4.30 mg. Ca pantothenate, 3 fig- vitamin Bi 2 , 5.5 mg. niacin, 25 mg. inositol, 500 I.U. vitamin A, 75 I.C.U. vitamin D3, 5 mg. a-tocopheryl acetate and 160 mg. choline chloride. 2 Furnished the following amounts per 100 gms. of diet: 1.00 gm. KC1, 0.75 gm. iodized salt, 0.35 gm. MgS0 4 , 36 mg. MnS0 4 -H 2 0, 36 mg. FeSCv 7H 2 0, 19.2 mg. ZnC0 3 , 2 mg. CuS0 4 -5H 2 0, 0.34 mg. NaMo0 4 -2H 2 0, 0.3 mg. CoCl 2 -6H 2 0 and 0.28 mg. KI.
TABLE 2.—Composition of basal diet, experiment 3 Ingredients Ground yellow corn Soybean oil meal (50% protein) Hydrogenated fat (Primex) Corn gluten meal Cond. fish solubles Alfalfa meal (17% protein, dehydrated) Iodized salt DL-methionine Vitamins1 MnSCvH 2 0, AR grade ZnC0 3 , AR grade Cellulose and supplements Protein, % Metabolizable energy, Cal./lb. ME/P Phosphorus, % analyzed Calcium, % analyzed
% 55.60 28.50 3.00 3.00 2.00 1.50 0.25 0.075 0.05 0.02 0.0125 6.00 21.8 1,360 62/1 0.37 0.17
1 Furnished the following per 100 gms. of diet: 0.275 mg. riboflavin, 0.33 mg. menadione sodium bisulfite, 0.44 mg. Ca pantothenate, 0.66 mg. niacin, 55 mg. choline chloride, 0.88 jug- vitamin B12, 220 I.U. vitamin A, 50 I.C.U. vitamin D 3 and 0.4 mg. a-tocopheryl acetate.
Downloaded from http://ps.oxfordjournals.org/ at Kainan University on April 25, 2015
in contrast to the "replacement" technique. Day and Hill (1959) reported that soft phosphate could replace 25 to 50% of supplementary defluorinated phosphate in the diets of turkeys to 20 weeks of age. Baruah et al. (1960b) using the "replacement" technique concluded that soft phosphate could replace a portion of the dicalcium phosphate or defluorinated phosphate in chick diets. Because the comparative availabilities of combinations of phosphates have not been determined, it was desirable to ascertain to what extent the chick could utilize these combinations. It was also desirable to know if biological values of phosphates determined with a purified diet could be applied to practical diets.
1323
AVAILABILITY OF PHOSPHORUS
The procedure for determining the availability of experimental phosphates using a standard curve has been reported by Gillis et al. (1954). These workers also described the method for preparation of the bones for ashing. RESULTS AND DISCUSSION
Experiment 1 was conducted to determine the bioligical availability of Curacao phosphate and soft phosphate when fed alone and in combination. I t was necessary to add 0.15% phosphorus from betatricalcium phosphate to the diets in which soft phosphate was tested in order to obtain good livability. Previous results (Table 3) obtained in this laboratory showed that an insufficient number of chicks survived the three-week experimental period when soft phosphate was the only source of supplemental phosphorus. Good livability is necessary to get a valid measure of the biological avail-
TABI.E 3. —Evidence of the low availability of phosphorus in soft phosphate cSupplement i „ i
Phosphorus fc
No. of
.Befa-tricalcium phosphate
0.20 0.25 0.30 0.35
0 0 0 0
Soft phosphate
0.25 0.30 0.35 0.40
90 91 82 60
samples
Av. mor-
taUty
%
ability of the phosphorus in any phosphate source. The results of experiment 1 are given in Table 4. The comparative availability of the phosphorus in Curacao phosphate was 6 8 % . These results agreed with those of Scott et al. (1956). These investigators showed t h a t poults could utilize commercial dicalcium phosphate more efficiently than Curacao phosphate. Curacao phosphate was utilized approximately as well as reagent grade dicalcium phosphate which has been shown to be 7 0 % available to chicks when compared to a betatricalcium phosphate standard. The biological value of soft phosphate was 36. The effect of 0.15% phosphorus supplied by ieto-tricalcium phosphate was eliminated from the calculation by subtracting it from the total phosphorus obtained with the standard curve. The availability of the phosphorus in a mixture containing equal amounts of phosphorus from Curacao phosphate and soft phosphate was 4 2 % . The average biological value of the individual phosphates was 52. The results of this experiment showed t h a t the availability of a combination of phosphates was no better than the average of their individual availabilities. The second experiment was conducted to determine if the results obtained in the
Downloaded from http://ps.oxfordjournals.org/ at Kainan University on April 25, 2015
Peterson X C o b b s White Plymouth Rock male chicks. The experimental period was 4 weeks. All chicks were reared in pens with raised wire mesh floors. H e a t was supplied by electrically heated brooders. Feed and distilled water were supplied ad libitum. The chicks were weighed weekly and feed consumption records were maintained. Percentage bone ash was the criterion used to determine the biological availability of the various phosphates by the chicks. The reference phosphate was betatricalcium phosphate. The standard curve was the regression line obtained by plotting the percentage bone ash obtained at the various levels of added phosphorus against the logarithm of the added phosphorus. The regression line was calculated using the method described by Hegsted (1948) and Almquist (1953).
1324
T. S. N E L S O N AND H. T.
PEELER
TABLE 4. -Biological availability of single and combined phosphates, experiment 1 Biological value
0.20 0.25 0.30 0.35
140 160 166 170
35.57 41.25 43.43 46.16
100
Curacao phosphate (CP) 1
0.30 0.35
144 167
35.55 42.59
68
Soft phosphate (SP)2
0.45 1 0.552
142 159
41.83 41.57
36
CP+SP3
0.60 0.80
148 160
42.35 42.85
42
Be/a-tricalcium phosphate
Supplemental phosphorus
%
Actual
Calculated
%
%
52
1 This diet contained 0.15% phosphorus from Seta-tricalcium phosphate and 0.30% phosphorus from soft phosphate. 2 This diet contained 0.15% phosphorus from Seta-tricalcium phosphate and 0.40% phosphorus from soft phosphate. 3 Each supplement provided 50% of the added phosphorus.
first experiment were reproducible. Different samples of Curacao phosphate and soft phosphate were used in this experiment. In addition, dicalcium phosphate was also tested. Again, as in the first experiment, the diets used to determine the biological value of soft phosphate contained 0.15% phosphorus from beta-tricalcium phosphate. The results of this experiment are given in Table 5. The biological value of dicalcium phosphate was 97 and the values obtained for Curacao phosphate and soft phosphate were in close agreement with those found in the first experiment. The available phosphorus in combinations of these phosphates, with each supplement supplying an equal amount of phosphorus, was very close to the calculated value. The calculated biological values of these phosphate mixtures were made using the averages of the individual phosphates. This experiment confirmed the findings in the first experiment. Poor quality phosphates were not improved when fed as mixtures with higher quality phosphates. The third experiment was conducted to
determine if the comparisons of biological availability obtained when the purified diet was fed could be applied to a test using a practical type diet. The experimental plan and results are shown in Table 6. The biological values of the individual phosphates agreed very closely with those obtained in experiment 2. Thus, the biological values of phosphates obtained with the purified diet can be applied to practical diets. Combinations of these phosphates, when each supplement provided the same proportion of phosphorus, had biological values approximately equal to the averages of the biological values of the supplements tested. Therefore, no synergistic effect of phosphates was observed in this experiment. In order to obtain valid comparisons of phosphates, it is necessary t h a t (1) a sensitive criterion of measure be used, (2) a critically phosphorus deficient diet be fed, (3) the levels of added phosphorus not be in excess of the animal's requirement, and (4) a suitable standard be used for comparison to the phosphates tested. No phosphate, regardless of how good or
Downloaded from http://ps.oxfordjournals.org/ at Kainan University on April 25, 2015
Bone ash
%
Av. wt. gain, gm.
Supplement.
1325
AVAILABILITY OF P H O S P H O R U S
TABLE 5.—Biological availability of single and combined phosphates, experiment Z Biological value
Bone ash
%
Av. wt. gain, gm.
0.20 0.25 0.30 0.35
134 142 169 159
29.71 34.88 39.73 41.76
100
Dicalcium phosphate (DCP)
0.25 0.35
142 168
33.98 41.34
97
Curacao phosphate (CP)
0.30 0.40
144 157
32.63 33.91
68
Soft phosphate (SP)
0.45i 0.55 2
133 131
34.66 36.60
31
DCP+CP3
0.30 0.40
150 156
33.90 38.41
77
82
DCP+SP3
0.40 0.60
133 156
35.73 46.11
68
64
CP+SP3
0.60 0.80
139 138
36.64 38.98
42
49
DCP+CP+SP"
0.45 0.60
142 154
38.41 42.72
63
65
Supplement
Seto-tricalcium
phosphate
Supplemental phosphorus
%
Actual
%
Calculated
%
how poor it may be, can be evaluated when tested in diets containing excess phosphorus. D a y and Hill (1959) fed diets to turkeys which, in terms of biologically available phosphorus, contained phosphorus in excess of the recommendations by the National Research Council (1954) and b y Scott (1960). The available phosphorus contents of the diets containing soft phosphate were calculated assuming this supplement to be only 2 5 % available. When 5 0 % of the phosphorus from defluorinated rock phosphate was replaced by the phosphorus in soft phosphate, the available phosphorus contents of these diets were still in excess of the turkey's requirements for this mineral, based on the National Research Council
recommendations. Based on the recommendations b y Scott (1960) the starter diet was borderline deficient in phosphorus and the remaining diets contained an excess of this mineral. T h e phosphorus replaced in these diets was merely excess phosphorus. No true evaluation of soft phosphate was actually determined. Baruah et al. (1960a) calculated the availability of a defluorinated phosphate and soft phosphate by dividing the percentage bone ash obtained with the test phosphate by the percentage bone ash obtained with the standard phosphate. The respective availabilities of these phosphates were reported as 95.5 and 85.7%. According to this method, the availability of no added phosphorus in the basal diet
Downloaded from http://ps.oxfordjournals.org/ at Kainan University on April 25, 2015
1 This diet contained 0.15% phosphorus from fteto-tricalcium phosphate and 0.30% phosphorus from soft phosphate. 2 This diet contained 0.15% phosphorus from ftcto-tricalcium phosphate and 0.40% phosphorus from soft phosphate. 3 Each supplement provided 50% of the added phosphorus. 4 Each supplement provided 33.3% of the added phosphorus.
1326
T. S. NELSON AND H. T. PEELER TABLE 6.—Biological availability of single and combined, phosphates, experiment 3 Phosphorus
Supplement
Biologi cal value
Av. wt. gain, gm.
Bone ash
Perosis
%
%
Actual Calculated
Added,
Available
%
%
0.04 0.08 0.12 0.16 0.20
0.04 0.08 0.12 0.16 0.20
281 313 305 332 357
32.20 35.69 40.73 42.80 44.13
50 10 0 0 0
100
DCP 2
0.05 0.15
0.05 0.15
301 336
33.56 41.62
44 0
99
CP 3
0.10 0.15
0.07 0.10
295 345
36.11 38.87
33 20
69
SP"
0.20 0.30
0.07 0.11
298 328
35.77 40.06
67 10
36
DCP+CP5
0.10 0.20
0.08 0.17
331 352
36.01 43.60
20 10
83
84
DCP+SP6
0.10 0.20
0.07 0.15
317 352
36.56 41.92
30 10
74
68
CP+SP5
0.10 0.20
0.06 0.11
324 310
33.94 40.14
30 40
57
53
DCP+CP+SPS
0.15 0.225
0.10 0.16
323 354
38.20 42.78
33 30
69
68
B-TCP 1
3 4 6 6
%
.Beta-tricalcium phosphate. Dicalcium phosphate. Curacao phosphate. Soft phosphate. Each supplement provided 50% of the added phosphorus. Each supplement provided 33.3% of the added phosphorus.
was 79.9%. The critical, measurable range was actually 79.9-100%. The increases in bone ash attributed to soft phosphate and defluorinated phosphate were then 5.8/20.1 and 15.6/20.1, respectively. The availabilities of these phosphates, based on a scale of 0-100% would be 29 and 78% respectively. Baruah et al. (1960b) in their studies on combinations of soft phosphate with other phosphates fed a basal diet which was not critically deficient in phosphorus nor was a sensitive criterion used. Moreover, the diet used contained a low level of energy, thereby decreasing the range of response to supplemental phosphorus. The requirement of available phosphorus, to produce
maximum bone ash, for chicks fed this diet would be approximately 0.36% based on the results of Peeler et al. (1960). Since the chick reaches maximum weight before attaining maximum bone ash, the requirement for growth on this diet would be less than 0.36% available phosphorus. Therefore, these phosphates, and combinations of them, were not tested over a critical range. The total phosphorus content of poultry rations is meaningless because the chick can utilize phytin phosphorus only to a limited degree (Gillis et al., 1957). The question of whether to express the usable phosphorus in such rations as available phosphorus or inorganic phosphorus oc-
Downloaded from http://ps.oxfordjournals.org/ at Kainan University on April 25, 2015
1 2
%
1327
AVAILABILITY OF PHOSPHORUS
44
/?-Tricalciam Phosphate Dicalcium Phosphate Curacao Phosphate Soft Phosphate
42
I+ff i+m E+m
40
38 x z o o
36
34
"
32 0-01
0O2
0-04
O06
LOGARITHM OF ADDED AVALIABLE
0 0 8 0-10
0-20
PH0SPH0RUS,%
FIG. 1. The relationship of biologically available phosphorus to percentage bone ash.
casionally occurs. The inaccuracy of the term "inorganic phosphate" is clearly demonstrated by the results given in Tables 3, 4 and 5. The entire phosphorus contents of these diets used in these experiments were inorganic. Varying results are obtained at the different levels of the phosphates, depending on the quality of the individual phosphates. However, when these phosphates are expressed on the basis of biologically available phosphorus they contained, the results agreed more closely. The advantage of expressing phosphorus as available phosphorus instead of inorganic phosphorus is also shown in Table 6. Varying results were obtained at specific levels of inorganic phosphorus. However, when expressed in terms of available phosphorus, excellent correlation existed between available phosphorus added and percentage bone ash. This is demonstrated in Figure 1.
The advantage of using percentage bone ash as the criterion of response instead of gain is also demonstrated. The percentage perosis observed in this experiment decreased as the percentage bone ash increased indicating that strong bones are needed to prevent this condition. The highest incidence of perosis was observed in the chicks fed diets containing soft phosphate. SUMMARY
Experiments were conducted with chicks fed purified and practical type diets to determine the biological availability of single and combined phosphate sources. When determined with a purified diet using 6eto-tricalcium phosphate as the reference material, the biological value of one sample of dicalcium phosphate was 97%. The average biological values of two samples each of Curacao phosphate and
Downloaded from http://ps.oxfordjournals.org/ at Kainan University on April 25, 2015
< UJ
1328
T . S. N E L S O N AND H . T. P E E L E R
soft phosphate were 68 a n d 3 4 % respectively.
The
biological
values
of
these
phosphates determined with a practical diet were almost identical with those obtained with the purified diet. Therefore, the results obtained with a purified diet can be applied to practical type diets. When either diet was fed, the biological value of combinations of phosphates was no better t h a n the sum of the available phosphorus contributed b y each source.
Available phosphorus is a better expression
of
the
phosphorus
content
of
poultry rations t h a n either total or inorganic phosphorus. REFERENCES Almquist, H. J., 1953. Evaluation of vitamin requirement data. Poultry Sci. 32: 122-128. Baruah, J. N., R. E. Davis, B. L. Reid and J. R. Couch, 1960a. Phosphorus availability from the ash of unidentified factor sources. Poultry Sci. 39: 840-842. Baruah, J. N., R. E. Davis, B. L. Reid and J. R. Couch, 1960b. Utilization of phosphorus from defluorinated and colloidal phosphate by chicks and laying hens. Poultry Sci. 39: 843-849. Day, E. J., and J. E. Hill, 1959. Zinc, potassium and certain other minerals as additives to practical
Studies with Corn-Soya Laying Diets 4. ENVIRONMENTAL TEMPERATURE—A FACTOR AFFECTING PERFORMANCE OF PULLETS FED DIETS SUBOPTIMAL IN PROTEIN D . J. BRAY AND J. A. G E S E L L
Department of Animal Science, University of Illinois, Urbana, Illinois (Received for publication December 5, 1960)
I
N
OUR
experience,
egg
production
trends of pullets fed diets suboptimal in
peratures was possible. No such effect was observed
when
the
diets
contained
a
protein have varied with season when the
plethora of protein. Conceivably, with a
birds were confined to a building where
plethora of protein in the diet, a reduction
only limited control over prevailing tem-
in the v o l u n t a r y consumption of feed does
Downloaded from http://ps.oxfordjournals.org/ at Kainan University on April 25, 2015
No synergistic effect of such phosphate combinations occurred.
turkey starter and grower rations. Poultry Sci. 38: 646-649. Gillis, M. B., L. C. Norris and G. F. Heuser, 1954. Studies on the biological value of inorganic phosphates. J. Nutrition, 52: 115-126. Gillis, M. B., K. W. Keane and R. A. Collins, 195V. Comparative metabolism of phytate and inorganic P 32 by chicks and poults. J. Nutrition, 62: 13-26. Hegsted, D. M., 1948. The determination of minimum vitamin requirements for growth. J. Nutrition, 35: 399-409. Motzok, I., D. Arthur and H. D. Branion, 1956. Utilization of phosphorus from various phosphate supplements by chicks. Poultry Sci. 35: 627-649. National Research Council, 1954. Nutrient requirements for domestic animals. I. Nutrient requirements for poultry. Peeler, H. T., T. S. Nelson and N. Storer, 1960. The effect of energy on the phosphorus requirement of the chick. Poultry Sci. 39: 1282. Scott, M. L., H. E. Butters and G. O. Ranet, 1956. Studies on bone formation in turkey poults. Proc. Cornell Nutr. Conf. p. 102-109. Scott, M. L., 1960. Turkey rations. New York State College of Agriculture Extension Stencil No. 208. Wilcox, R. A., C. W. Carlson, W. Kohlmeyer and G. G. Gas tier, 1954. The availability of phosphorus from different sources for poults fed purified diets. Poultry Sci. 33: 1010-1014. Wilcox, R. A., C. W. Carlson, W. Kohlmeyer and G. G. Gastler 1955. The availability of phosphorus from different sources for poults fed practical-type diets. Poultry Sci. 34: 1017-1023.