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Calcification in the Poult
Calcification in the Poult JAMES C. FRITZ, JOHN H. HOOPER, AND HOWARD P. MOORE Borden's Nutritional Research Laboratory, Elgin, Illinois (Received for publication, November 27, 1944)
PREVIOUS report from this laboratory (Willgeroth, Halpin, Halloran, and Fritz, 1944) suggested the use of turkey poults for the assay of vitamin D in poultry feed supplements. Using various samples, good agreement was obtained between assays using chicks and assays using poults. Reports by Bird (1944), Boucher (1944), and Sanford and Jukes (1944) indicated that not all sources of vitamin D were equally effective for poults when compared on the basis of A.O.A.C. chick units. Investigation of the diets used indicated that there were large differences in the mineral contents. This study was undertaken to determine the effect of the levels and ratio of calcium and phosphorus upon calcification in the poult. Special attention was given to the possibility that these elements might have an influence upon the relative effectiveness of vitamin D from different sources. MATERIALS AND METHODS
The poults used in this study were hatched at the laboratory from the experimental flock of bronze turkeys. All work was done in electrically heated, thermostatically controlled, screen floored starting batteries. The groups were relatively small, consisting in most cases of seven to twelve poults. Except as modifications were indicated, the experimental procedure followed the technic recommended in the A.O.A.C. Methods of Analysis (1940). In some tests the toe ash method of Baird and MacMil. Ian (1942) was used to supplement the tibiae ash data. The toe ash method was
also used with ten groups of poults which were subsequently saved for other purposes. Later growth of the poults was fully equal to that obtained with other poults from the same stock which had all their toes intact. The vitamin D supplements were carefully evaluated by repeated A.O.A.C. chick vitamin D assays. The tuna livers were a blend of various samples. This blend contained 26 percent solids and 452 A.O.A.C. units of vitamin D per gram. The reference cod liver oil was from fresh samples used within 60 days of the date of distribution by the U.S.P. Committee. Two samples of pure vitamin D3 were used. One was a solution, prepared by Dr. Huber of the Winthrop Chemical Company, and used soon after receipt at this laboratory. Repeated chick tests, including comparison against several lots of U.S.P. reference cod liver oil, indicated the unexpectedly high potency of 59,490,000 A.O.A.C. units of vitamin D per gram of D 3 . The other vitamin D3 sample was the crystalline vitamin received over three years ago. This material had been stored in a refrigerator at approximately 0° C. during most of this period, but it had been opened and portions removed at various times during the three years. This material was found by repeated chick tests to contain 57,900,000 A.O.A.C. units of vitamin D per gram. It may be concluded that there was little or no deterioration of this sample during the storage period. In fact, this activity is somewhat greater than that originally found in this sample. On the other hand, the possibility of deteri-
324
Downloaded from http://ps.oxfordjournals.org/ at University of Hawaii, PBRC, Kewalo Marine Lab. on June 3, 2015
A
325
CALCIFICATION IN THE POULT
used in these tests is shown in Table 1. Table 2 shows the calcium and phosphorus content of these diets. Differences between these data and those given in the original reports based on similar diets may be ascribed to variations in the lots of ingredients used by the several laboratories. TABLE 2.—Calcium and phosphorus
content of the diets Diet Borden Bird Boucher Sanford-Jukes
Percent calcium
Percent phosphorus
1.51 1.15 1.19 1.92
1.00 0.78 0.58 1.01
EXPERIMENTAL
DATA
Table 3 summarizes the effect of varying the calcium and phosphorus content of the ratio when the level of vitamin D was kept constant at 70 A.O.A.C. units per 100 grams and tuna livers were used as the source of the vitamin D.
TABLE 1.—Composition of Diets Boucher Sanford & Jukes
Ingredient
Borden
Bird
Yellow corn Barley Wheat Wheat bran Middlings Oats Casein Flaydry* Dried skim milk Sardine meal Soybean meal Dedhydrated alfalfa Dried brewer's yeast Meat & bone scrap Calcium phosphate St. bone meal Calcium carbonate Salt with I 2 & Mn Vegetable oil
48.0
29.0
25.9
—: —
— —
— —
:
10.0 10.0
—
15.0 5.0
10.0 10.0 10.0 6.0
—
—
— — —. —
8.0
8.0
—
5.0 17.5
12.0
—
12.0
—
5.0
5.0
7.5
7.5
2.0
6.0
6.0
.—
— — —
,
10.0 10.0 10.0 6.0
25.0 20.0 7.0 15.0
1.0
—
—
—
2.0
1.5
—
—
_ —
—
1.0
1.0
2.0
1.5
1.0
0.5 2.0
0.6' 2.0
0.5
—
1.0
—
Poultry feed supplement contg. whey solubles.
TABLE 3.—Effect of varying mineral content with constant vitamin D intake Boucher's diet less CaCOs plus tuna livers to supply 70 AOAC Units vitamin D/100 Gms. Supplement to diet None 1% CaC0 3 2 % CaC0 3 3 % CaCOs 4 % CaCOa 2 % Bone meal 2%CaC03+2%Bone 4 % C a C 0 3 + 2 % Bone 6%CaC03+2%Bone 3 % C a C 0 3 + 3 % Bone
%Ca
%P
0.35 0.74 1.14 1.53 1.92 0.93 1.72 2.50 3.28 2.39
0.60 0.60 0.59 0.59 0.58 0.87 0.87 0.87 0.87 1.00
% Bone Wt. ash gain 22.08 24.12 26.92 34.73 38.41 28.88 33.53 40.27 43.21 38.43
gms. 67 100 99 160 141 76 100 124 133 110
In Table 4, average weights at 6 weeks of age and percentage toe ash at 3 weeks of age were obtained for a comparison of vitamin D 3 , reference cod liver oil, and tuna livers. The activity of reference cod liver oil, pure D3, and dihydrotachysterol was
Downloaded from http://ps.oxfordjournals.org/ at University of Hawaii, PBRC, Kewalo Marine Lab. on June 3, 2015
oration of the U.S.P. reference cod liver oil number 2 cannot be ignored. The dihydrotachysterol used in these tests was a carefully standardized oil solution prepared by Dr. Huber. Assays at this laboratory indicated that the pure sterol contained 78,333 U.S.P. units of vitamin D and 176,000 A.O.A.C. units of vitamin D per gram. The U.S.P. potency agreed well with data given by McChesney (1943), but the A.O.A.C. potency was only about one-half that reported by McChesney. The D-activated animal sterol was a sample of powdered Delsterol which was found to contain 2,495 A.O.A.C. units of vitamin D per gram. The basal diets used in this study were modeled after those used earlier by this laboratory (1944), by Bird (1944), by Boucher (1944), and by Sanford and Jukes (1944). The composition of each diet as
326
J A M E S C. F R I T Z , J O H N H . H O O P E R , AND H O W A R D P . M O O R E TABLE 4.—Comparison of D3, ref. C.L.O-,
and tuna livers on growth of poults AOAC Units D/100 Gms.
Supplement to Borden diet None
0
6.79
67
16.62 17.28 17.62 11.70 14.40 14.94 16.35 17.85 16.80
104 149 45 70 45 70 100 200
A
^ f
All dead gms. 695 741 739 602 712 633 726 743 807
measured with observations including weight gains during the three week period, percentage tibiae ash, a n d percentage toe ash. These data are summarized in Table 5. TABLE 5.—Activity of ref. C.L.O., D3, and
dihydrotachysterol when fed to poults
Supplement to Borden diet None USP Ref. C.L.O. USP Ref. C.L.O. USP Ref. C.L.O. Ds Ds Ds Dihydrotachysterol Dihydrotachysterol Dihydrotachysterol
A hat3wks D/100 % ? -Wt'. Gms. T l b l a e T o e s gain
0 45 60 75 67 89 111 22 29 37
26.65 34.42 38.70 41.96 43.81 47.35 46.41 29.02 29.15 29.84
7.83 12.60 14.62 15.84 14.22 14.70 16.96 8.71 9.19 10.41
gms. 86 137 138 150 135 144 150 61 81 81
TABLE 6.—Comparative effectiveness of ref.
C.L.O. and D-activated animal sterols for poults Supplement to Sanford & Jukes diet None USP Ref. C.L.O. USP Ref. C.L.O. USP Ref. C.L.O. D-activated sterols D-activated sterols
AOAC Units % Ash at 3 wks. wAv.' D/100 Tibiae Toes w t ' gain Gms. 0 80 160 240 75 100
36.26 42.57 44.99 46.77 44.76 47.27
11.13 15.49 17.37 16.50 16.71 18.26
gms. 89 147 152 109 141 99
T h e effect of adding calcium to t h e ration is indicated in the three experiments summarized in Tables 7, 8, a n d 9. DISCUSSION I t is obvious from t h e d a t a presented t h a t minerals h a d a pronounced effect upon both growth and calcification in poults. T h e d a t a in Table 3 indicate t h a t the calcium had a growth promoting value which phosphorus did n o t duplicate. Both were effective in producing better calcification. T h e d a t a seems to indicate t h a t higher than normal levels of both calcium
TABLE 7.—Influence of minerals on relative effectiveness of Dz and tuna livers. Supplement to Borden diet less 1% CaC03 CaCOs Source of D 0 0 0 0 3% 3% 3% 3% 5%
'
D3 D3 Tuna Tuna D3 . D3 Tuna Tuna Tuna
livers livers livers livers livers
AOAC Units D/100 Gms.
% Ash, Tibiae
3wk. Toes
Av. Wt. gain
89 149 60 100 89 149 60 100 60
40.68 43.53 36.56 38.77 46.15 46.30 44.60 46.53 42.35
15.34 17.28 11.96 13.23 18.53 19.18 16.55 18.99 17.68
gms. 77 84 35 105 88 143 97 126 161
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D3 Ds D3 USP Ref. C.L.O. USP Ref. C.L.O. Tuna livers Tuna livers Tuna livers Tuna livers'
% Toe ash
Table 6 is essentially confirmation of the work reported b y Sanford a n d Jukes (1944). I t is interesting to note t h a t this was the only one of the four diets studied which failed t o produce severe rickets when not supplemented with a source of vitamin D .
327
CALCIFICATION IN THE POULT TABLE 8.—Influence of minerals on relative effectiveness of ref. C.L.O. and Z>s Supplements to Bird's diet CaCOs Source of D 0 0 2% 2%
AOAC Units D/100 Gms.
% Ash, Tibiae
3wk. Toes
Av. Wt. gain
80 119 80 119
32.11 38.44 38.01 43.44
11.01 13.43 14.65 19.07
gms. 95 106 161 148
USP Ref. C.L.O. LV USP Ref. C.L.O. D3
Supplements to Borden diet less 1 % CaCOs Source of D
CaCOs
A.O.A.C Units D/100 Gms.
% Tibiae ashat3wks.
,
> 0 0 0 0 3% 3% 3% 3%
-
USP USP D3 D3 USP USP D3 Ds
Ref. C.L.O. Ref. C.L.O. Ref. C.L.O. Ref. C.L.O.
and phosphorus are needed for optimum calcification with low levels of vitamin D. .Best results were obtained with rather wide low levels of vitamin D. Best results were obtained with rather wide calcium rphosphorus ratios. Some of the ratios which gave superior results with poults were so wide that they would have been expected to increase the vitamin D requirements of chicks, according to Hart, Scott, Kline, and Halpin (1930), Wilgus (1931), and numerous other investigators. At suboptimal levels, growth varied with the vitamin D intake. It may be noted that the tuna livers, and at times the reference cod liver oil, had a growth promoting value which the D3 and the D -activated animal sterol apparently did not possess. This effect can scarcely be attributed to the vitamin A carried along with the fish sources. The vitamin A levels were believed to be more than adequate, since the alfalfa used in the basal diets contained a minimum of 83,500 U.S.P. units of vitamin A activity per pound at
60 100 87 145 60 100 87 145
29.53 36.26 40.66 42.10 37.89 45.36 40.93 45.18
Av. Wt. gain gms. 127 125 120 120 156 150 107 130
the time the diets were mixed. Furthermore, additional carotene in oil was added to the diets used in the experiment summarized in Table 8. Here, too, the growth promoting effect of reference cod liver oil Was apparent. The reference cod liver oil seemed peculiarly ineffective when used to supplement rations low in calcium and phosphorus. With low levels of minerals each of the other supplements appeared to be more effective. The D 3 and the Delsterol seemed most effective of the' supplements when used with low mineral rations, followed closely in most cases by the tuna livers. With ample calcium and phosphorus in the ration, satisfactory calcification was obtained with nominal levels of any of the supplements tested. Only with suboptimal levels of minerals was there any advantage in feeding more than 100 A.O.A.C. units of vitamin D per 100 grams of ration. It seems apparent that the poor calcification obtained in many instances by Bird (1944) and by Boucher (1944) might
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TABLE 9.—Influence of minerals on effectiveness of iref. C.L.Q. and A
328
JAMES C. FRITZ, JOHN H. HOOPER, AND HOWARD P. MOORE
1. Minerals have a marked influence on calcification in the poult. The calcium and phosphorus requirements for optimum calcification with low vitamin D intake are relatively high, and a wide ratio seems optimum. 2. Reference cod liver oil seems peculiarly ineffective when used to supplement low mineral rations.
3. When used to supplement rations containing the usual practical mineral levels, 70 to 100 A.O.A.C. units of vitamin D per 100 grams of ration, from any source, seems to be ample for satisfactory calcification in the poult. REFERENCES
Association of Official Agricultural Chemists, 1940. Official and Tentative Methods of Analysis. Sth ed., Washington, D. C. Baird, F. D., and M. J. MacMillan, 1942. Use of toes rather than tibiae in the A.O.A.C. chick method ' of vitamin D determination. Jour. A.O.A.C. 25: 518-524. Bird, H. R., 1944. Comparison of response of turkey poults and of chicks to different forms of vitamin D. J. Nutrition 27:377-383. Boucher, R. V., 1944. Efficacy of vitamin D from different sources for turkeys. J. Nutrition 27:403413. Hart, E. B., H. T. Scott, O. L. Kline, and J. G. Halpin, 1930. The calcium phosphorus ratio in the nutrition of the growing chick. Poultry Sci. 9:296306. McChesney, E. W., 1943. Studies of calcium and phosphorus metabolism in the chick. 1. The comparative effect of vitamins D2 and D 3 and dihydrotachysterol given orally and intramuscularly. J. Nutrition 26:81-94. Sanford, T. D., and T. H. Jukes, 1944. Further observations on the vitamin D requirement of turkey poults. Poultry Sci. 23:221-223. Wilgus, H. S., 1931. The quantitative requirements of the growing chick for calcium and phosphorus. Poultry Sci. 10:108-117. Willgeroth, G. B., J. L. Halpin, H. R. Halloran, and J. C. Fritz, 1944. Use of turkeys for assay of vitamin D. Jour. A.O.A.C. 27:289-295.
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be due to the relatively low calcium and phosphorus levels in their diets. It is, however, difficult to understand why there should have been such an extreme difference in response when D 3 was used instead of fish oil. In our study the same trend is noted but the quantitative differences are not as great. It should also be noted that the mineral content of the Sanford and Jukes diet (1944) was fully as high as that used in the Borden diet. Apparent differences between cases where a vitamin D source was comparatively effective in the presence of suboptimal mineral content and cases where it was comparatively ineffective, include differences in the fat content of the diet. On this basis, the tuna livers would be expected to act more nearly like the D 3 and the D-activated animal sterols. Such does seem to be the case. Further work along this line is planned and will be started as soon as a supply of poults can be assured. CONCLUSIONS
PREVIOUS report from this laboratory (Willgeroth, Halpin, Halloran, and Fritz, 1944) suggested the use of turkey poults for the assay of vitamin D in poultry feed supplements. Using various samples, good agreement was obtained between assays using chicks and assays using poults. Reports by Bird (1944), Boucher (1944), and Sanford and Jukes (1944) indicated that not all sources of vitamin D were equally effective for poults when compared on the basis of A.O.A.C. chick units. Investigation of the diets used indicated that there were large differences in the mineral contents. This study was undertaken to determine the effect of the levels and ratio of calcium and phosphorus upon calcification in the poult. Special attention was given to the possibility that these elements might have an influence upon the relative effectiveness of vitamin D from different sources. MATERIALS AND METHODS
The poults used in this study were hatched at the laboratory from the experimental flock of bronze turkeys. All work was done in electrically heated, thermostatically controlled, screen floored starting batteries. The groups were relatively small, consisting in most cases of seven to twelve poults. Except as modifications were indicated, the experimental procedure followed the technic recommended in the A.O.A.C. Methods of Analysis (1940). In some tests the toe ash method of Baird and MacMil. Ian (1942) was used to supplement the tibiae ash data. The toe ash method was
also used with ten groups of poults which were subsequently saved for other purposes. Later growth of the poults was fully equal to that obtained with other poults from the same stock which had all their toes intact. The vitamin D supplements were carefully evaluated by repeated A.O.A.C. chick vitamin D assays. The tuna livers were a blend of various samples. This blend contained 26 percent solids and 452 A.O.A.C. units of vitamin D per gram. The reference cod liver oil was from fresh samples used within 60 days of the date of distribution by the U.S.P. Committee. Two samples of pure vitamin D3 were used. One was a solution, prepared by Dr. Huber of the Winthrop Chemical Company, and used soon after receipt at this laboratory. Repeated chick tests, including comparison against several lots of U.S.P. reference cod liver oil, indicated the unexpectedly high potency of 59,490,000 A.O.A.C. units of vitamin D per gram of D 3 . The other vitamin D3 sample was the crystalline vitamin received over three years ago. This material had been stored in a refrigerator at approximately 0° C. during most of this period, but it had been opened and portions removed at various times during the three years. This material was found by repeated chick tests to contain 57,900,000 A.O.A.C. units of vitamin D per gram. It may be concluded that there was little or no deterioration of this sample during the storage period. In fact, this activity is somewhat greater than that originally found in this sample. On the other hand, the possibility of deteri-
324
Downloaded from http://ps.oxfordjournals.org/ at University of Hawaii, PBRC, Kewalo Marine Lab. on June 3, 2015
A
325
CALCIFICATION IN THE POULT
used in these tests is shown in Table 1. Table 2 shows the calcium and phosphorus content of these diets. Differences between these data and those given in the original reports based on similar diets may be ascribed to variations in the lots of ingredients used by the several laboratories. TABLE 2.—Calcium and phosphorus
content of the diets Diet Borden Bird Boucher Sanford-Jukes
Percent calcium
Percent phosphorus
1.51 1.15 1.19 1.92
1.00 0.78 0.58 1.01
EXPERIMENTAL
DATA
Table 3 summarizes the effect of varying the calcium and phosphorus content of the ratio when the level of vitamin D was kept constant at 70 A.O.A.C. units per 100 grams and tuna livers were used as the source of the vitamin D.
TABLE 1.—Composition of Diets Boucher Sanford & Jukes
Ingredient
Borden
Bird
Yellow corn Barley Wheat Wheat bran Middlings Oats Casein Flaydry* Dried skim milk Sardine meal Soybean meal Dedhydrated alfalfa Dried brewer's yeast Meat & bone scrap Calcium phosphate St. bone meal Calcium carbonate Salt with I 2 & Mn Vegetable oil
48.0
29.0
25.9
—: —
— —
— —
:
10.0 10.0
—
15.0 5.0
10.0 10.0 10.0 6.0
—
—
— — —. —
8.0
8.0
—
5.0 17.5
12.0
—
12.0
—
5.0
5.0
7.5
7.5
2.0
6.0
6.0
.—
— — —
,
10.0 10.0 10.0 6.0
25.0 20.0 7.0 15.0
1.0
—
—
—
2.0
1.5
—
—
_ —
—
1.0
1.0
2.0
1.5
1.0
0.5 2.0
0.6' 2.0
0.5
—
1.0
—
Poultry feed supplement contg. whey solubles.
TABLE 3.—Effect of varying mineral content with constant vitamin D intake Boucher's diet less CaCOs plus tuna livers to supply 70 AOAC Units vitamin D/100 Gms. Supplement to diet None 1% CaC0 3 2 % CaC0 3 3 % CaCOs 4 % CaCOa 2 % Bone meal 2%CaC03+2%Bone 4 % C a C 0 3 + 2 % Bone 6%CaC03+2%Bone 3 % C a C 0 3 + 3 % Bone
%Ca
%P
0.35 0.74 1.14 1.53 1.92 0.93 1.72 2.50 3.28 2.39
0.60 0.60 0.59 0.59 0.58 0.87 0.87 0.87 0.87 1.00
% Bone Wt. ash gain 22.08 24.12 26.92 34.73 38.41 28.88 33.53 40.27 43.21 38.43
gms. 67 100 99 160 141 76 100 124 133 110
In Table 4, average weights at 6 weeks of age and percentage toe ash at 3 weeks of age were obtained for a comparison of vitamin D 3 , reference cod liver oil, and tuna livers. The activity of reference cod liver oil, pure D3, and dihydrotachysterol was
Downloaded from http://ps.oxfordjournals.org/ at University of Hawaii, PBRC, Kewalo Marine Lab. on June 3, 2015
oration of the U.S.P. reference cod liver oil number 2 cannot be ignored. The dihydrotachysterol used in these tests was a carefully standardized oil solution prepared by Dr. Huber. Assays at this laboratory indicated that the pure sterol contained 78,333 U.S.P. units of vitamin D and 176,000 A.O.A.C. units of vitamin D per gram. The U.S.P. potency agreed well with data given by McChesney (1943), but the A.O.A.C. potency was only about one-half that reported by McChesney. The D-activated animal sterol was a sample of powdered Delsterol which was found to contain 2,495 A.O.A.C. units of vitamin D per gram. The basal diets used in this study were modeled after those used earlier by this laboratory (1944), by Bird (1944), by Boucher (1944), and by Sanford and Jukes (1944). The composition of each diet as
326
J A M E S C. F R I T Z , J O H N H . H O O P E R , AND H O W A R D P . M O O R E TABLE 4.—Comparison of D3, ref. C.L.O-,
and tuna livers on growth of poults AOAC Units D/100 Gms.
Supplement to Borden diet None
0
6.79
67
16.62 17.28 17.62 11.70 14.40 14.94 16.35 17.85 16.80
104 149 45 70 45 70 100 200
A
^ f
All dead gms. 695 741 739 602 712 633 726 743 807
measured with observations including weight gains during the three week period, percentage tibiae ash, a n d percentage toe ash. These data are summarized in Table 5. TABLE 5.—Activity of ref. C.L.O., D3, and
dihydrotachysterol when fed to poults
Supplement to Borden diet None USP Ref. C.L.O. USP Ref. C.L.O. USP Ref. C.L.O. Ds Ds Ds Dihydrotachysterol Dihydrotachysterol Dihydrotachysterol
A hat3wks D/100 % ? -Wt'. Gms. T l b l a e T o e s gain
0 45 60 75 67 89 111 22 29 37
26.65 34.42 38.70 41.96 43.81 47.35 46.41 29.02 29.15 29.84
7.83 12.60 14.62 15.84 14.22 14.70 16.96 8.71 9.19 10.41
gms. 86 137 138 150 135 144 150 61 81 81
TABLE 6.—Comparative effectiveness of ref.
C.L.O. and D-activated animal sterols for poults Supplement to Sanford & Jukes diet None USP Ref. C.L.O. USP Ref. C.L.O. USP Ref. C.L.O. D-activated sterols D-activated sterols
AOAC Units % Ash at 3 wks. wAv.' D/100 Tibiae Toes w t ' gain Gms. 0 80 160 240 75 100
36.26 42.57 44.99 46.77 44.76 47.27
11.13 15.49 17.37 16.50 16.71 18.26
gms. 89 147 152 109 141 99
T h e effect of adding calcium to t h e ration is indicated in the three experiments summarized in Tables 7, 8, a n d 9. DISCUSSION I t is obvious from t h e d a t a presented t h a t minerals h a d a pronounced effect upon both growth and calcification in poults. T h e d a t a in Table 3 indicate t h a t the calcium had a growth promoting value which phosphorus did n o t duplicate. Both were effective in producing better calcification. T h e d a t a seems to indicate t h a t higher than normal levels of both calcium
TABLE 7.—Influence of minerals on relative effectiveness of Dz and tuna livers. Supplement to Borden diet less 1% CaC03 CaCOs Source of D 0 0 0 0 3% 3% 3% 3% 5%
'
D3 D3 Tuna Tuna D3 . D3 Tuna Tuna Tuna
livers livers livers livers livers
AOAC Units D/100 Gms.
% Ash, Tibiae
3wk. Toes
Av. Wt. gain
89 149 60 100 89 149 60 100 60
40.68 43.53 36.56 38.77 46.15 46.30 44.60 46.53 42.35
15.34 17.28 11.96 13.23 18.53 19.18 16.55 18.99 17.68
gms. 77 84 35 105 88 143 97 126 161
Downloaded from http://ps.oxfordjournals.org/ at University of Hawaii, PBRC, Kewalo Marine Lab. on June 3, 2015
D3 Ds D3 USP Ref. C.L.O. USP Ref. C.L.O. Tuna livers Tuna livers Tuna livers Tuna livers'
% Toe ash
Table 6 is essentially confirmation of the work reported b y Sanford a n d Jukes (1944). I t is interesting to note t h a t this was the only one of the four diets studied which failed t o produce severe rickets when not supplemented with a source of vitamin D .
327
CALCIFICATION IN THE POULT TABLE 8.—Influence of minerals on relative effectiveness of ref. C.L.O. and Z>s Supplements to Bird's diet CaCOs Source of D 0 0 2% 2%
AOAC Units D/100 Gms.
% Ash, Tibiae
3wk. Toes
Av. Wt. gain
80 119 80 119
32.11 38.44 38.01 43.44
11.01 13.43 14.65 19.07
gms. 95 106 161 148
USP Ref. C.L.O. LV USP Ref. C.L.O. D3
Supplements to Borden diet less 1 % CaCOs Source of D
CaCOs
A.O.A.C Units D/100 Gms.
% Tibiae ashat3wks.
,
> 0 0 0 0 3% 3% 3% 3%
-
USP USP D3 D3 USP USP D3 Ds
Ref. C.L.O. Ref. C.L.O. Ref. C.L.O. Ref. C.L.O.
and phosphorus are needed for optimum calcification with low levels of vitamin D. .Best results were obtained with rather wide low levels of vitamin D. Best results were obtained with rather wide calcium rphosphorus ratios. Some of the ratios which gave superior results with poults were so wide that they would have been expected to increase the vitamin D requirements of chicks, according to Hart, Scott, Kline, and Halpin (1930), Wilgus (1931), and numerous other investigators. At suboptimal levels, growth varied with the vitamin D intake. It may be noted that the tuna livers, and at times the reference cod liver oil, had a growth promoting value which the D3 and the D -activated animal sterol apparently did not possess. This effect can scarcely be attributed to the vitamin A carried along with the fish sources. The vitamin A levels were believed to be more than adequate, since the alfalfa used in the basal diets contained a minimum of 83,500 U.S.P. units of vitamin A activity per pound at
60 100 87 145 60 100 87 145
29.53 36.26 40.66 42.10 37.89 45.36 40.93 45.18
Av. Wt. gain gms. 127 125 120 120 156 150 107 130
the time the diets were mixed. Furthermore, additional carotene in oil was added to the diets used in the experiment summarized in Table 8. Here, too, the growth promoting effect of reference cod liver oil Was apparent. The reference cod liver oil seemed peculiarly ineffective when used to supplement rations low in calcium and phosphorus. With low levels of minerals each of the other supplements appeared to be more effective. The D 3 and the Delsterol seemed most effective of the' supplements when used with low mineral rations, followed closely in most cases by the tuna livers. With ample calcium and phosphorus in the ration, satisfactory calcification was obtained with nominal levels of any of the supplements tested. Only with suboptimal levels of minerals was there any advantage in feeding more than 100 A.O.A.C. units of vitamin D per 100 grams of ration. It seems apparent that the poor calcification obtained in many instances by Bird (1944) and by Boucher (1944) might
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TABLE 9.—Influence of minerals on effectiveness of iref. C.L.Q. and A
328
JAMES C. FRITZ, JOHN H. HOOPER, AND HOWARD P. MOORE
1. Minerals have a marked influence on calcification in the poult. The calcium and phosphorus requirements for optimum calcification with low vitamin D intake are relatively high, and a wide ratio seems optimum. 2. Reference cod liver oil seems peculiarly ineffective when used to supplement low mineral rations.
3. When used to supplement rations containing the usual practical mineral levels, 70 to 100 A.O.A.C. units of vitamin D per 100 grams of ration, from any source, seems to be ample for satisfactory calcification in the poult. REFERENCES
Association of Official Agricultural Chemists, 1940. Official and Tentative Methods of Analysis. Sth ed., Washington, D. C. Baird, F. D., and M. J. MacMillan, 1942. Use of toes rather than tibiae in the A.O.A.C. chick method ' of vitamin D determination. Jour. A.O.A.C. 25: 518-524. Bird, H. R., 1944. Comparison of response of turkey poults and of chicks to different forms of vitamin D. J. Nutrition 27:377-383. Boucher, R. V., 1944. Efficacy of vitamin D from different sources for turkeys. J. Nutrition 27:403413. Hart, E. B., H. T. Scott, O. L. Kline, and J. G. Halpin, 1930. The calcium phosphorus ratio in the nutrition of the growing chick. Poultry Sci. 9:296306. McChesney, E. W., 1943. Studies of calcium and phosphorus metabolism in the chick. 1. The comparative effect of vitamins D2 and D 3 and dihydrotachysterol given orally and intramuscularly. J. Nutrition 26:81-94. Sanford, T. D., and T. H. Jukes, 1944. Further observations on the vitamin D requirement of turkey poults. Poultry Sci. 23:221-223. Wilgus, H. S., 1931. The quantitative requirements of the growing chick for calcium and phosphorus. Poultry Sci. 10:108-117. Willgeroth, G. B., J. L. Halpin, H. R. Halloran, and J. C. Fritz, 1944. Use of turkeys for assay of vitamin D. Jour. A.O.A.C. 27:289-295.
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be due to the relatively low calcium and phosphorus levels in their diets. It is, however, difficult to understand why there should have been such an extreme difference in response when D 3 was used instead of fish oil. In our study the same trend is noted but the quantitative differences are not as great. It should also be noted that the mineral content of the Sanford and Jukes diet (1944) was fully as high as that used in the Borden diet. Apparent differences between cases where a vitamin D source was comparatively effective in the presence of suboptimal mineral content and cases where it was comparatively ineffective, include differences in the fat content of the diet. On this basis, the tuna livers would be expected to act more nearly like the D 3 and the D-activated animal sterols. Such does seem to be the case. Further work along this line is planned and will be started as soon as a supply of poults can be assured. CONCLUSIONS