Phosphorus in the Nutrition of the Adult Hen1

Phosphorus in the Nutrition of the Adult Hen 1 2. THE RELATIVE AVAILABILITY OF PHOSPHORUS FROM SEVERAL SOURCES FOR CAGED LAYERS E . P . SlNGSEN, L . D . MATTERSON, J . J . TLUSTOHOWICZ AND W . J . PUDELKIEWICZ Poultry Science Department, The University of Connecticut, Storrs, Connecticut 06268 (Received for publication August 13, 1968)

T HAS long been recognized that marked differences exist in availability to the chick of the phosphorus occurring in both natural and synthetic compounds. Relatively little is known, however, about phosphorus availability for the adult hen. Common (1939) reported that much of the phosphorus in feces from pullets was in the form of phytin. Gillis et al. (1953) concluded that hens utilized the phosphorus from calcium phytate to a greater extent than did chicks, but only one half as effectively as that from dicalcium phosphate or defluorinated phosphate. Walter and Aitken (1962) reported that caged hens fed a diet supplemented with dicalcium phosphate required 0.60% total and a calculated 0.36% nonphytin phosphorus to support maximum egg production over an 11-month period. Hens utilized phosphorus from sodium or calcium phytate more effectively than chicks, according to Maddaiah et al. (1963), but both compounds were less effective than dicalcium phosphate. Waldroup et al. (1967) reported that the phosphorus in hominy feed did not support normal egg production, but that it did aid in preventing the development of embryonic rickets. In an extensive review on the utilization of phytate phosphorus by both chicks and hens, Nelson (1967) concurred with the present recommendation of the Committee on 1 Scientific Contribution No. 330, Agricultural Experiment Station, University of Connecticut, Storrs.

Animal Nutrition, National Academy of Sciences, National Research Council (NAS-NRC) (I960), that 30% of the total phosphorus from plant feed ingredients be considered available. In sharp contrast, Nott et al. (1967) presented data based on balance studies indicating that hens fed 2.5% and 3.5% calcium excreted 84% and 94% respectively of their phytate phosphorus intake in the droppings, and they concluded that the phosphorus requirements of poultry should be expressed in terms of nonphytate phosphorus. The objective of this report is to present biological assay data on the ability of the hen to utilize phosphorus from several sources. MATERIALS AND METHODS

The simplified low phosphorus basal ration (0.20% total P) was the same as previously described by Singsen et al. (1962), with the exception of the addition of 0.05% DL-methionine to the ration. In experiment 1, each replicate consisted of 15 range-reared Darby S.C. White Leghorn pullets housed in individual 11"X18" cages. In experiment 2, 30 Arbor Acres Leghorn-type pullets, housed two in a cage, comprised each replicate. All treatments were replicated twice in each experiment. Data were compiled for ten 28day periods on egg production, efficiency of feed utilization, body weights, egg weights, and mortality. There was a moderate, evenly distributed mortality from pickouts when birds were housed two in a

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S I N G S E N , M A T T E R S O N , TLUSTOHOWICZ AND P U D E L K I E W I C Z

100%. All materials of less than kilogram quantities were weighed to t h e nearest gram on a Toledo scale. EXPERIMENTAL AND DISCUSSION

Experiments 1 and 2 were identical in design. A bioassay standard curve, based on egg production, was developed b y supplementing t h e basal ration with commercial dicalcium phosphate so as to achieve total phosphorus levels of 0.20, 0.34, 0.48, and 0.62%. T h e increment size, 0.14%, was chosen for two reasons: (1) exchanging all the degermed, debranned white corn meal in the basal ration for ground yellow corn added exactly 0.14% total phosphorus, thus permitting a direct assessment of t h e relative value of t h e total phosphorus contained in corn and, (2) 0.34% total phosphorus is a suboptimal level of this mineral for caged layers and therefore is in a highly desirable range in which to test the relative availability of the phosphorus from various sources. Furthermore, at this level the supplement contributed 4 1 . 2 % of t h e total phosphorus, which should be sufficient to reveal differences in availability if they existed. An outline of the experimental plan and details of diet composition are shown in Table 1. Although t h e patterns of results were

TABLE 1.—Experimental design and ration composition for experiments 1 and 2

%

Total phosphorus, Added phosphorus , Total calcium, %

%

0 .20 0 3. 00

0 .34 0..14 3..00

0.48 0.28 3.00

0.62 0.42 3.00

0 .34 0 .14 3 ,00

0 .34 0..14 3..00

0 .34 0 .14 3 .00

0 .34 0 .14 3 .00

28.077

28.077 0.765

28.077 1.530

28.077 2.295

28.077

28.077

28.077

28.077

Variables, % Basal diet Dicalcium phosphate 1 Ground yellow corn Defluorinatd rock phosphate 2 Curacao rock phosphate 3 Soft phosphate 4 Ground limestone5 White corn meal

Totals 1 2 3 4 5

67.288 0.791 0.981 4.635 67.288

100.0

4.218 66.940

100.0

3.804 66.589

100.0

3.387 66.241

100.0

4.635

100.0

Dynafos. International Minerals and Chemical Company, Inc., Skokie, Illinois. Multifos. International Minerals and Chemical Company, Inc., Skokie, Illinois. Curacao Island rock phosphate. Mixture of three open market samples. Soft phosphate with colloidal clay. Soft Phosphate Research Institute, Inc., Ocala, Florida. Degermed and debranned. The Quaker Oats Company, Inc., Chicago, Illinois.

3.994 67.138

100.0

3.755 67.187

100.0

1.493 3.938 66.492

100.0

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cage and, since this was not related to the treatments being studied, it was compiled separately and eliminated from the mortality d a t a presented here. B o t h experiments were initiated when the pullets reached 5 0 % to 7 0 % egg production. T h e y were conducted in a heated, forceventilated, windowless room with 16 hours of light per day. Feed and water were supplied ad libitum. Four feed ingredients, ground yellow corn, a defluorinated rock phosphate, Curacao rock phosphate, and soft phosp h a t e were studied as phosphorus sources of unknown availability. For each experiment, several bags of each inorganic phosphorus supplement were obtained, blended, analyzed for phosphorus, and then used through the experiment. T h e finished diets were also analyzed periodically for calcium and phosphorus. Phosphorus was determined according to the method of Fiske and Subbarow (1925) and calcium by the A.O.A.C. (1950) method. T h e samples were solubilized b y the wet oxidation method of Reitz et al. (1960). T h e d a t a were examined b y analysis of variance according to the method of Snedecor (1956). Calcium was maintained at 3.00% in all treatments b y varying the quantity of ground limestone, a n d corn was adjusted slightly to bring all diets to

389

PHOSPHORUS FOR THE H E N TABLE 2.—The relative effectiveness of phosphorus from several sources for caged layers (Experiment 1, combined data, 2 replicates)

cu

Source of phosphorus 1 Total phosphorus Added phosphorus

Basal 0.20 0

Average egg production, % (hen-day basis) Mortality, % 2 Feed consumed, g m s . / h e n / d a y Average final live weight, gms. Gain in live weight, gms. Egg weight, gms. 28 days 280 days

46.2

61.7

69.1

72.9

59.8

65.4

69.1

61.7

40.1 88 1,813 -12

13.3 106 2,076 223

3.3 106 1,991 235

6.7 107 2,103 284

20.0 103 1,873 57

10.0 105 2,085 268

20.0 104 2,069 175

6.7 100 1,904 80

58.2 63.4

58.9 66.4

58.8 65.9

58.1 65.0

58.5 63.8

58.2 65.0

58.4 65.0

57.2 63.0

DC 0.34 0.14

DC 0.48 0.28

DC 0.62 0.42

YC 0.34 0.14

DP 0.34 0.14

0.34 0.14

SP 0.34 0.14

very similar in experiments 1 and 2, there were significant differences (P<0.05) between years in both egg production and mortality, and therefore the results have been presented separately in Tables 2 and 3. The bioassay standard curves, based on final average egg production (hen-day basis, 40 weeks), and the relative egg production response to phosphorus from other sources are shown graphically in Figures 1 and 2. In both experiments, egg production increased markedly with the addition of 0.14% phosphorus from dicalcium phosphate, and then continued up at a slower rate to the maximum addition of 0.42% (0.62% total) fed. The addition of 0.14% phosphorus from the four supplements under test supported egg production responses that were similar to and not sta-

tistically different (P>0.05) from that obtained with dicalcium phosphate. Percent mortality decreased with increasing phosphorus from dicalcium phosphate, reaching its lowest point at 0.48% (total) in experiment 1 and 0.62% (total) in experiment 2. In both experiments, hens receiving supplemental phosphorus from a defluorinated rock phosphate sustained mortality similar to that occurring in the control group fed dicalcium phosphate. Birds fed soft phosphate exhibited a low mortality in experiment 1 and high mortality in experiment 2. Of interest was the higher mortality that occurred in the groups receiving supplemental phosphorus from either yellow corn or Curacao rock phosphate, particularly in view of the fact that both sources supported a marked increase in egg production. Although these

TABLE 3.—The relative effectiveness of phosphorus from several sources for caged layers (Experiment 2, combined data, 2 replicates) Source of phosphorus 1 Total phosphorus, % Added phosphorus, %

Basal 0.20 0

Average egg production, % (hen-day basis) Mortality, % 2 Feed consumed, g m s . / h e n / d a y Average final live weight, gms. Gain in live weight, gms. Egg weight, gms. 28 days 280 days

47.2

72.4

75.7

78.1

68.3

74.4

72.5

68.6

50.0 76 1,586 -79

18.3 96 1,845 225

15.0 97 1,908 207

3.3 98 1,850 187

28.3 96 1,770 95

18.3 95 1,868 199

28.3 93 1,768 112

26.6 96 1,721 83

57.3 59.6

58.4 62.5

58.3 63.8

59.5 62.6

59.8 65.6

57.1 63.0

58.3 63.7

59.3 61.0

1 2

See key at bottom of Table 2. Exclusive of pickouts.

DC 0.34 0.14

DC 0.48 0.28

DC 0.62 0.42

YC 0.34 0.14

DP 0.34 0.14

CU 0.34 0.14

SP 0.34 0.14

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-Basal ration. DC—Basal ration plus dicalcium phosphate. YC—Basal ration with yellow corn replacing the degermed, debranned white corn. DP—Basal ration plus defluorinated rock phosphate. CU—Basal ration plus Curacao Island rock phosphate. SP—Basal ration plus soft phosphate. 2 Exclusive of pickouts.

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SlNGSEN, MATTERSON, T L T J S T O H O W I C Z AND PuDELKIEWICZ

• -DICALCIUM PHOSPHATE x- -YELLOW CORN 0 -DEFLUORINATED ROCK PHOSPHATE i -CURACAO ROCK PHOSPHATE 9-SOFT PHOSPHATE 0-30

0.40

0.50

0.60

0.70

PERCENT PHOSPHORUS

FIG. 1. The effect of phosphorus source and level on egg production. Expt. 1.

differences were not statistically significant at this point ( P > 0 . 0 5 ) , subsequent experiments to be reported separately showed one of them (C.R.P.) to be both consistent and significant. I n both experiments hens fed the basal ration were unable to maintain body weight. T h e addition of 0.14% or more phosphorus from either dicalcium phosphate or the defluorinated rock phosphate supported maximum gain in live weight. Hens receiving 0.14% supplemental phosphorus from either yellow corn or soft phosphate made relatively small gains in live weight, while those receiving Curacao rock phosphate showed an intermediate gain in weight. The differences in gain in live weight cannot be explained on the basis of either feed intake or total phosphorus intake, since feed consumption, and hence phosphorus intake, were very uniform for all treatments. Egg weight was reduced on the basal diet and in the groups receiving soft phosphate, b u t was not consistently affected by any of the other treatments. T h e finding was not unexpected t h a t the total phosphorus from yellow corn was relatively effective in supporting egg production. Several reports, Pepper et al. (1959) (floor-housed birds), Walter and Aitken (1962), and O'Rourke et al. (1955)

One example of differential usefulness

I a o

• -DICALCIUM PHOSPHATE *-YELLOW CORN o -DEFLUORINATED ROCK PHOSPHATE 0-CURACAO POCK PHOSPHATE • -SOFT PHOSPHATE 020

0.30

0-40

0.50

060

070

PERCENT PHOSPHORUS

FIG. 2. The effect of phosphorus source and level on egg production. Expt. 2.

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0.20

(caged birds) have indicated t h a t practical diets, in which the phosphorus came primarily from plant products, supported fair to good egg production. Furthermore, Maddaiah et al. (1963) concluded, in agreement with Gillis et al. (1953), t h a t hens utilized isolated p h y t a t e phosphorus more efficiently than did chicks. Taking the average egg production of the basal groups in experiments 1 and 2 as zero, 0.14% supplemental phosphorus from yellow corn increased egg production 17.4 percentage points in comparison with 20.4 percentage points for the same quantity from dicalcium phosphate, giving a relative effectiveness of 8 5 . 3 % . According to Singsen (1945), more than 8 0 % of the phosphorus in yellow corn is in the form of phytic acid or its salts. I t is obvious therefore that, in the experiments reported here, much of it was used for egg production. The failure of the phosphorus from yellow corn to reduce mortality or support gain in live weight as effectively as t h a t from dicalcium phosphate suggests either a qualitative differential in usefulness for different biological functions or a sequence differential in which one function takes precedence over another in the presence of a suboptimal supply of this nutrient.

P H O S P H O R U S FOR THE

T h e conclusion of N o t t et al. (1967) t h a t hens fed adequate calcium levels utilize little, if any, of their dietary p h y t a t e phosphorus appears to be in conflict with the d a t a reported here. I t is the opinion of the authors, however, t h a t the explanation lies somewhere in the mechanism by which phytic acid phosphorus is digested, metabolized, and excreted rather than in a true conflict of results. T h e simple fact that a similar quantity of p h y t a t e phosphorus can be measured in both the feed and the feces does not prove t h a t it has passed through the body in a totally inactive state. On the contrary, its high degree of effectiveness in supporting egg production, as reported here, indicates an active biological role prior to its excretion in the feces. D a t a reported by Singsen et al. (1950)

391

and Gillis et al. (1957) utilizing P 32 labeled phytates, demonstrated that p h y t a t e phosphorus is absorbed into the body. Exchange reaction has been postulated as the mechanism involved, b u t it is possible t h a t the process is much more complex. According to Sebrell and Harris (1954), numerous esters of inositol, involving both organic moieties and minerals other than phosphorus, calcium, and magnesium occur in plants, animals, and bacteria. T h e y are easily synthesized and degraded under the influence of enzymes from these sources. Maddaiah et al. (1964) demonstrated, in vitro, phytase activity in homogenates from the intestine of laying hens. Rapoport (1940) reported t h a t phytic acid occurs in the nucleated erythrocytes of the chicken, thus indicating t h a t it is either absorbed intact or synthesized in avian tissue. I t seems possible t h a t partial hydrolysis could occur in the intestinal tract with the phosphorus being temporarily replaced by some other moiety. This would account for the appearance of P 32 labeled p h y t a t e phosphorus in the body and also for its apparent ability to support both growth and egg production. Upon reaching the lower intestinal tract which, according to M a y n a r d and Loosli (1962) is the major route of phosphorus excretion, the intermediary compounds of inositol could easily be resynthesized to phytic acid under the influence of phytase from either the intestine or bacteria. In the presence of excess calcium, the phytic acid would be precipitated and excreted as phytin or calcium p h y t a t e . In agreement with H a r m s et al. (1961), the use of soft phosphate as the source of supplemental phosphorus resulted in egg production equivalent to t h a t obtained with dicalcium phosphate. There was, however, a reduced gain in live weight and a slightly lower egg weight. No explanation is available for the wide variation in

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involving phytin phosphorus, reported previously by Singsen (1945) and Vandepopuliere et al. (1961), was based on the observation that, in chicks, plant source phosphorus was capable of supporting growth b u t contributed little or nothing to increasing bone ash. I t is possible t h a t its inability to contribute to bone mineralization was a major factor in the increased hen mortality reported here. On the basis of d a t a presented here, it is apparent that the phosphorus in yellow corn is substantially more available to hens for egg production than the 3 0 % figure currently recommended by the NASN R C Committee on Animal Nutrition. One should recognize, however, t h a t it must be supplemented with nonphytin phosphorus in order to achieve optimum overall performance. I t is also apparent t h a t this high relative availability m a y apply only to yellow corn, since Gillis et al. (1953), working with wheat bran, and Waldroup et al. (1967), working with hominy feed, obtained relatively poor results from the phosphorus in those ingredients.

HEN

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SINGSEN, MATTEESON, TLUSTOHOWICZ AND PUDELKIEWICZ

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mortality between experiments 1 and 2. ucts of unknown availability contributing It would appear that the phosphorus in 0.14% (41.2%) of the total. this product is biologically less effective Rate of egg production on a hen-day for adult hens than that in dicalcium basis was not significantly different on any phosphate. of the phosphorus sources, although it The egg production and egg weight data tended to be slightly lower when the phosobtained from hens receiving supplemen- phorus source was yellow corn. Gain in tal phosphorus from Curacao rock phos- body weight was markedly less when the phate were equivalent to those obtained supplemental phosphorus came from yelwith dicalcium phosphate. Gain in live low corn or soft phosphate, intermediate body weight was intermediate, however, with Curacao rock phosphate, and normal and mortality averaged 8.35% higher on the defluorinated rock phosphate. Effithan similar groups fed dicalcium phos- ciency of feed utilization for egg producphate. On the basis of all performance tion was not influenced by phosphorus characteristics, it would appear that the source. phosphorus in Curacao rock phosphate is Mortality was consistently higher when biologically less effective for adult hens hens received either yellow corn or Curathan that in dicalcium phosphate but cao rock phosphate as their source of supslightly more effective than that in yellow plemental phosphorus than when the othcorn or soft phosphate. er supplements were fed. In both experiments, 0.62% total ACKNOWLEDGMENTS (0.52% nonphytin or available) phosphorus from the basal diet and dicalcium The authors are indebted to Mr. Peter phosphate supported the highest egg pro- McManus for technical assistance with duction. This finding is very close to the diet preparation and care of the experi0.55% available phosphorus requirement mental birds, to Mrs. Joseph Lucas for aid reported previously for caged layers by on the statistical analyses, and to Mr. J. Singsen el al. (1962). The combined data A. Serafin for assistance with the first exfrom the lots fed supplemental phosphorus periment. from dicalcium phosphate indicate that The trace mineral mixture used in these minimum mortality also occurred at experiments was contributed by the Lime0.62% total (0.52% nonphytin) phospho- stone Products Corporation of America, rus. Newton, New Jersey. Both financial and/or product contributions were reSUMMARY ceived at various times from the InternaThe relative biological usefulness of tional Minerals and Chemical Corporaphosphorus from several sources for caged tion, Skokie, Illinois; H. J. Baker and layers has been explored. Two biological Bro., New York; the Soft Phosphate Reassay experiments of 40 weeks' duration search Institute, Ocala, Florida; and the utilized dicalcium phosphate to develop a Central Connecticut Cooperative Farmers standard curve based on egg production, Association, Manchester, Connecticut. and included a defluorinated rock phosThe senior author is also indebted to phate, Curacao rock phosphate, soft phos- Dr. R. H. Harms, Poultry Science Departphate, and yellow corn as phosphorus ment, University of Florida, Gainesville, sources of unknown availability. Total di- Florida, for both critical evaluation and etary phosphorus was maintained at a the office facilities used during the prepsuboptimal level, 0.34%, with the prod- aration of this manuscript.

PHOSPHORUS FOR THE H E N REFERENCES

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Association of Official Agricultural Chemists, 1950. Methods of Analysis. Washington, D. C. Common, R. H., 1939. Phytic acid and mineral metabolism in poultry. Nature, 143: 379-380. Fiske, E. H., and Y. Subbarow, 1925. The colormetric determination of phosphorus. J. Biol. Chem. 66: 375. Gillis, M. B., L. C. Norris and G. F. Heuser, 1953. Phosphorus metabolism and requirements of hens. Poultry Sci. 32: 977-984. Gillis, M. B., K. W. Keane and R. A. Collins, 1957. Comparative metabolism of phytate and inorganic P32 by chicks and poults. J. Nutr. 62: 13-26. Maddaiah, V. T., B. J. Hulett, A. A. Kurnick and B. L. Reid, 1963. Availability of calcium phytate, sodium phytate, and dicalcium phosphate in chicks, hens, and rats. Poultry Sci. 42: 1286. Maynard, L. A., and J. K. Loosli, 1962. Animal Nutrition. McGraw-Hill Book Co., Inc., New York, New York. National Academy of Sciences-National Research Council, 1960. Nutrient requirements for domestic animals. 1. Nutrient requirements for poultry. Nelson, T. S., 1967. The utilization of phytate phosphorus by poultry—a review. Poultry Sci. 46: 862-871. Nott, H., T. R. Morris and T. G. Taylor, 1967. Utilization of phytate phosphorus by laying hens and young chicks. Poultry Sci. 46: 1301. O'Rourke, W. F., P. H. Phillips and W. W. Cravens, 1955. The phosphorus requirements of growing chicks and laying pullets fed practical rations. Poultry Sci. 34: 47-54. Pepper, W. F., S. J. Slinger, J. D. Summers and G. C. Ashton, 1959. On the phosphorus require-