- Email: [email protected]
Further Observations on the Vitamin D Requirement of Turkey Poults
Further Observations on the Vitamin D Requirement of Turkey Poults T. D. SANFORD AND T. H. JUKES From the Biological Laboratories, F. E. Booth Company, Emeryville, California and the Division of Poultry Husbandry, University of California, Davis, California (Received for publication November 19, 1943)
EXPERIMENTAL
Day-old turkey poults were placed in electrically heated battery brooders in a room from which all daylight was excluded, and were fed the experimental diets immediately. Each group contained 14 birds, 7 of which were obtained from eggs laid by the University of California experimental flock (Jukes and Sanford, 1939) and 7 from a commercial hatchery. Two basal diets were used. Diet 1 contained ground yellow corn, 25 parts; ground barley 20, ground wheat 7, wheat bran 15, sardine meal 17.5, dried skimmilk 5, dehydrated alfalfa meal 7.5, ground limestone 1.5, steamed bone meal 1, salt 0.5. Diet 2 contained ground yellow corn 36 parts; wheat flour middlings 22, commercial casein 18, ground whole oats 11, dried brewers' yeast 4, dried whey 4,
ground limestone 2, steamed bone meal 2, salt 1. The sources of vitamin D were as follows: (1) U.S.P. Reference Cod Liver Oil number 2. (2) A sample of fish oil blend ("Sardilene 400") which was assayed with chicks, TABLE 1.—Chick vitamin D assay of fish oil blend. Nineteen chicks were used in each group Supplement per 100 grams diet*
None 5 units 1 7.5 units 1 10 units 1 12.5 units 1 15 units 1 17.5 units 1 20 units 1 18.8 milligrams 25 milligrams 31.2 milligrams 37.5 milligrams
2 2 2 2
Composite bone ash at 31 days
Estimate of potency
Percent 33.9 36.6 38.4 41.9 42.7 43.7 44.5 46.1 39.1 39.9 42.4 43.6 Average
Units per gram
448 372 406 392 404
* Supplement 1 = reference cod liver oil no. 2, U.S.P.; supplement 2 = fish oil blend.
as shown in Table 1, and was found to have a value of 404 A.O.A.C. units of vitamin D per gram. (3) A sample of an irradiated animal sterol product ("Delsterol") which was assayed with chicks. Five separate assays in three different laboratories gave a mean value of 496 A.O.A.C. units of vitamin D per milligram. One of the chick assays is summarized, as an example, in Table 2.
[221]
Downloaded from http://ps.oxfordjournals.org/ at Univ of Iowa-Law Library on June 19, 2015
T T WAS reported previously (Jukes and -*• Sanford, 1939) that approximately 200 A.O.A.C. chick units of vitamin D per 100 grams of diet were required by turkey poults in the first 4 weeks to produce maximal calcification. Attention was drawn to the observation that turkeys and chicks might exhibit a species difference in their response to different forms of vitamin D. This species difference has been further established by the work of Willgeroth and co-workers with dihydrotachysterol (1943). The present report deals with further experiments on the response of poults to varying levels of vitamin D.
222
T. D. SANFORD AND T. H. JUKES
TABLE 2.—Example of vitamin D assay of the activated animal sterol used in this investigation; according to Methods of Analysis, Association of Official Agricultural Chemists, 1940, p. 371
TABLE 3.—Effect Group No.
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16
Basal Diet
2 2
of various sources and levels of vitamin D on turkey poults Vitamin D Supplement*
Weight at 4 weeks
Crooked Breastbones at 4 weeks
Ash in Tibiae
Per 100 grams diet None None 40 units 1 80 units 1 120 units 1 160 units 1 200 units 1 240 units 1 80 units 2 140 units 2 200 units 2 80 units 3 140 units 3 200 units 3 None 80 units 1
Grams 173 163 196 277 297 265 279 284 253 315 309 273 300 275 185 328
Percent 100 100 45 42 8 33 0 0 21 17 0 9 0 13 100 45
Percent 29.6 30.2 37.7 43.4 46.2 46.4 49.4 49.7 43.2 49.2 49.2 48.4 49.5 50.1 29.8 40.4
* Vitamin D Supplement 1 =• Reference Cod Liver Oil No. 2, U.S.P.; 2 = fish oil blend; 3 = activated animal sterol.
Downloaded from http://ps.oxfordjournals.org/ at Univ of Iowa-Law Library on June 19, 2015
figures in the last column represent the average values for the left and right tibiae of each entire group. The bone ash values of the left tibiae are Composite' Supplement per 100 grams of diet compared in Table 4, group by group, for bone ash turkeys of the two different sources. Were Percent it not for the excessive difference in group 30.0 None 31.4 4 units Reference Cod Liver Oil 9, the difference between the bone ash 33.3 4 units Reference Cod Liver Oil values of the poults from source 1 and 37.1 8 units Reference Cod Liver Oil 36.1 8 units Reference Cod Liver Oil source 2 would be highly significant in 41.7 12 units Reference Cod Liver Oil groups 1 to 11, and the differences be40.6 12 units Reference Cod Liver Oil 35.1 .0107 milligrams Delsterol tween the bone ash values produced by 37.6 .0171 milligrams Delsterol vitamin D, fish oil blend and vitamin D, 39.0 .0213 milligrams Delsterol activated sterol would be significant. 38.4 .0213 milligrams Delsterol 42.1 .0256 milligrams Delsterol However, the differences between the 44. .0320 milligrams Delsterol bone ash values of poults from source 1 From these, and other results it was estimated and source 2 are not of sufficient magnithat one milligram of Delsterol contained 475 tude to indicate the need for a revision of A.O.A.C. units of vitamin D . previous conclusions (Jukes and Sanford, The turkeys were weighed weekly. At 1939) with respect to the level of vitamin four weeks of age they were examined for D needed by young turkeys. crooked breastbones and killed. The tibiae The data in Table 3 tend to confirm the were removed, cleaned, extracted and previous findings (Jukes and Sanford, ashed as described previously (Jukes and 1939). With Reference cod liver oil no. 2 Sanford, 1939). Ash determinations were (U.S.P.), a level of 200 units per 100 grams made upon the left tibiae individually and of diet 1 was required for maximum calciupon the right tibiae in groups. The re- fication at four weeks. The fish oil blend sults are summarized in Table 3. The and the activated animal sterol both ap-
F U R T H E R OBSERVATIONS ON V I T A M I N D R E Q U I R E M E N T OF T U R K E Y P O U L T S
223
TABLE 4.-—Comparison
of bone ash of turkeys from the University of California flock (source 1) with those of turkeys from a commercial hatchery (source 2) on identical diets
Group no.
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
Bone ashisource 1 29.5 29.7 38.5 43.4 46.6 46.3 49.0 49.8 45.2 48.8 49.4 47.4 49.1 49.9 29,0 41.3 Bone ash source 2 29.1 29.6 34.6 42.8 45.9 45.9 48.2 48.8 40.0 48.4 48.7 48.3 49.6 50.2 29.9 39.5
Diet 2, which was similar to t h a t of H a m m o n d (1941), failed to give maximum calcification when supplemented with 80 ' A.O.A.C. units of vitamin D (U.S.P. reference cod liver oil no. 2) per 100 grams. This result is in contrast with t h a t reported by H a m m o n d . I n one of the abstracts issued b y the Wistar I n s t i t u t e on M a r c h 15, 1944, which describe articles to appear in the Journal of Nutrition for M a y 1944, it is stated b y R. V. Boucher t h a t "if cod liver oil and sardine oil are assumed to be representative of fish oils, 200 chick units per 100 gm. of diet from such sources m a y be considered an approximation of the 'requirement' of poults. I n contrast, approximately 80 units from the irradiated preparations would satisfy the requirem e n t . " This statement is in agreement with our previous finding (Jukes and Sanford (1939) and with the present report. I n another of these abstracts, issued simultaneously, it is stated b y H. R. Bird "As measured by effect on per cent ash in the fat-free, dried tibiae, a given number of A.O.A.C. chick units of vitamin D from irradiated 7-dehydrocholesterol and the irradiated animal sterol were, respec-
tively, 2.29 and 1.83 times as effective for poults as the same number of A.O.A.C. chick units from Reference Cod Liver Oil." This finding is similar to our results and to the statement by Boucher. There appeared to be a greater tendency for the development of crooked breastbones on lower levels of vitamin D . SUMMARY (1) The vitamin D requirement of turkey poults was further investigated b y the use of three different sources of vitamin D which had been biologically assayed with chicks. (2) T h e vitamin D requirement for maximum calcification a t four weeks was in the neighborhood of 200 A.O.A.C. units of Reference cod liver oil no. 2, U.S.P., b u t the corresponding requirement was lower in the case of the other two sources of vitamin D . (3) A high incidence of crooked breastbones a t four weeks of age was observed to be associated with low levels of vitamin D . ACKNOWLEDGEMENT The authors are indebted to Dr. J. J. Waddell for supplying the irradiated animal sterol ("Delsterol") and to Dr. V. S. Asmundson for examining the data statistically. REFERENCES
Carver, J. S., and M. Rhian, 1942. The vitamin D requirement of turkey poults. Poultry Sci. 21: 260-262. Hammond, J. C, 1941. The vitamin D requirement of turkey poults. Poultry Sci. 20:204-205. Jukes, T. H„ and T. D. Sanford, 1939. The vitamin D requirements of young turkeys. J. Nutrition 18:71-85. Willgeroth, G. B., J. L. Halpin, H. R. Halloran, and J. C. Fritz. Turkeys and the vitamin D assay. Paper presented before 100th meeting, American Chemical Society, Pittsburgh, 1943.
Downloaded from http://ps.oxfordjournals.org/ at Univ of Iowa-Law Library on June 19, 2015
peared to be more efficacious for turkeys than for chicks under these conditions. I t was reported by Carver and Rhian (1942) t h a t 80 A.O.A.C. chick units of "fortified cod liver oil" per 100 grams of feed supplied adequate vitamin D for turkey poults. This does not necessarily conflict with the present observations, for if "fortified cod liver oil" contained activated animal sterol, for example, as a fortification, such an oil would tend to be more efficacious than the U.S.P. reference cod liver oil for turkeys.
EXPERIMENTAL
Day-old turkey poults were placed in electrically heated battery brooders in a room from which all daylight was excluded, and were fed the experimental diets immediately. Each group contained 14 birds, 7 of which were obtained from eggs laid by the University of California experimental flock (Jukes and Sanford, 1939) and 7 from a commercial hatchery. Two basal diets were used. Diet 1 contained ground yellow corn, 25 parts; ground barley 20, ground wheat 7, wheat bran 15, sardine meal 17.5, dried skimmilk 5, dehydrated alfalfa meal 7.5, ground limestone 1.5, steamed bone meal 1, salt 0.5. Diet 2 contained ground yellow corn 36 parts; wheat flour middlings 22, commercial casein 18, ground whole oats 11, dried brewers' yeast 4, dried whey 4,
ground limestone 2, steamed bone meal 2, salt 1. The sources of vitamin D were as follows: (1) U.S.P. Reference Cod Liver Oil number 2. (2) A sample of fish oil blend ("Sardilene 400") which was assayed with chicks, TABLE 1.—Chick vitamin D assay of fish oil blend. Nineteen chicks were used in each group Supplement per 100 grams diet*
None 5 units 1 7.5 units 1 10 units 1 12.5 units 1 15 units 1 17.5 units 1 20 units 1 18.8 milligrams 25 milligrams 31.2 milligrams 37.5 milligrams
2 2 2 2
Composite bone ash at 31 days
Estimate of potency
Percent 33.9 36.6 38.4 41.9 42.7 43.7 44.5 46.1 39.1 39.9 42.4 43.6 Average
Units per gram
448 372 406 392 404
* Supplement 1 = reference cod liver oil no. 2, U.S.P.; supplement 2 = fish oil blend.
as shown in Table 1, and was found to have a value of 404 A.O.A.C. units of vitamin D per gram. (3) A sample of an irradiated animal sterol product ("Delsterol") which was assayed with chicks. Five separate assays in three different laboratories gave a mean value of 496 A.O.A.C. units of vitamin D per milligram. One of the chick assays is summarized, as an example, in Table 2.
[221]
Downloaded from http://ps.oxfordjournals.org/ at Univ of Iowa-Law Library on June 19, 2015
T T WAS reported previously (Jukes and -*• Sanford, 1939) that approximately 200 A.O.A.C. chick units of vitamin D per 100 grams of diet were required by turkey poults in the first 4 weeks to produce maximal calcification. Attention was drawn to the observation that turkeys and chicks might exhibit a species difference in their response to different forms of vitamin D. This species difference has been further established by the work of Willgeroth and co-workers with dihydrotachysterol (1943). The present report deals with further experiments on the response of poults to varying levels of vitamin D.
222
T. D. SANFORD AND T. H. JUKES
TABLE 2.—Example of vitamin D assay of the activated animal sterol used in this investigation; according to Methods of Analysis, Association of Official Agricultural Chemists, 1940, p. 371
TABLE 3.—Effect Group No.
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16
Basal Diet
2 2
of various sources and levels of vitamin D on turkey poults Vitamin D Supplement*
Weight at 4 weeks
Crooked Breastbones at 4 weeks
Ash in Tibiae
Per 100 grams diet None None 40 units 1 80 units 1 120 units 1 160 units 1 200 units 1 240 units 1 80 units 2 140 units 2 200 units 2 80 units 3 140 units 3 200 units 3 None 80 units 1
Grams 173 163 196 277 297 265 279 284 253 315 309 273 300 275 185 328
Percent 100 100 45 42 8 33 0 0 21 17 0 9 0 13 100 45
Percent 29.6 30.2 37.7 43.4 46.2 46.4 49.4 49.7 43.2 49.2 49.2 48.4 49.5 50.1 29.8 40.4
* Vitamin D Supplement 1 =• Reference Cod Liver Oil No. 2, U.S.P.; 2 = fish oil blend; 3 = activated animal sterol.
Downloaded from http://ps.oxfordjournals.org/ at Univ of Iowa-Law Library on June 19, 2015
figures in the last column represent the average values for the left and right tibiae of each entire group. The bone ash values of the left tibiae are Composite' Supplement per 100 grams of diet compared in Table 4, group by group, for bone ash turkeys of the two different sources. Were Percent it not for the excessive difference in group 30.0 None 31.4 4 units Reference Cod Liver Oil 9, the difference between the bone ash 33.3 4 units Reference Cod Liver Oil values of the poults from source 1 and 37.1 8 units Reference Cod Liver Oil 36.1 8 units Reference Cod Liver Oil source 2 would be highly significant in 41.7 12 units Reference Cod Liver Oil groups 1 to 11, and the differences be40.6 12 units Reference Cod Liver Oil 35.1 .0107 milligrams Delsterol tween the bone ash values produced by 37.6 .0171 milligrams Delsterol vitamin D, fish oil blend and vitamin D, 39.0 .0213 milligrams Delsterol activated sterol would be significant. 38.4 .0213 milligrams Delsterol 42.1 .0256 milligrams Delsterol However, the differences between the 44. .0320 milligrams Delsterol bone ash values of poults from source 1 From these, and other results it was estimated and source 2 are not of sufficient magnithat one milligram of Delsterol contained 475 tude to indicate the need for a revision of A.O.A.C. units of vitamin D . previous conclusions (Jukes and Sanford, The turkeys were weighed weekly. At 1939) with respect to the level of vitamin four weeks of age they were examined for D needed by young turkeys. crooked breastbones and killed. The tibiae The data in Table 3 tend to confirm the were removed, cleaned, extracted and previous findings (Jukes and Sanford, ashed as described previously (Jukes and 1939). With Reference cod liver oil no. 2 Sanford, 1939). Ash determinations were (U.S.P.), a level of 200 units per 100 grams made upon the left tibiae individually and of diet 1 was required for maximum calciupon the right tibiae in groups. The re- fication at four weeks. The fish oil blend sults are summarized in Table 3. The and the activated animal sterol both ap-
F U R T H E R OBSERVATIONS ON V I T A M I N D R E Q U I R E M E N T OF T U R K E Y P O U L T S
223
TABLE 4.-—Comparison
of bone ash of turkeys from the University of California flock (source 1) with those of turkeys from a commercial hatchery (source 2) on identical diets
Group no.
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
Bone ashisource 1 29.5 29.7 38.5 43.4 46.6 46.3 49.0 49.8 45.2 48.8 49.4 47.4 49.1 49.9 29,0 41.3 Bone ash source 2 29.1 29.6 34.6 42.8 45.9 45.9 48.2 48.8 40.0 48.4 48.7 48.3 49.6 50.2 29.9 39.5
Diet 2, which was similar to t h a t of H a m m o n d (1941), failed to give maximum calcification when supplemented with 80 ' A.O.A.C. units of vitamin D (U.S.P. reference cod liver oil no. 2) per 100 grams. This result is in contrast with t h a t reported by H a m m o n d . I n one of the abstracts issued b y the Wistar I n s t i t u t e on M a r c h 15, 1944, which describe articles to appear in the Journal of Nutrition for M a y 1944, it is stated b y R. V. Boucher t h a t "if cod liver oil and sardine oil are assumed to be representative of fish oils, 200 chick units per 100 gm. of diet from such sources m a y be considered an approximation of the 'requirement' of poults. I n contrast, approximately 80 units from the irradiated preparations would satisfy the requirem e n t . " This statement is in agreement with our previous finding (Jukes and Sanford (1939) and with the present report. I n another of these abstracts, issued simultaneously, it is stated b y H. R. Bird "As measured by effect on per cent ash in the fat-free, dried tibiae, a given number of A.O.A.C. chick units of vitamin D from irradiated 7-dehydrocholesterol and the irradiated animal sterol were, respec-
tively, 2.29 and 1.83 times as effective for poults as the same number of A.O.A.C. chick units from Reference Cod Liver Oil." This finding is similar to our results and to the statement by Boucher. There appeared to be a greater tendency for the development of crooked breastbones on lower levels of vitamin D . SUMMARY (1) The vitamin D requirement of turkey poults was further investigated b y the use of three different sources of vitamin D which had been biologically assayed with chicks. (2) T h e vitamin D requirement for maximum calcification a t four weeks was in the neighborhood of 200 A.O.A.C. units of Reference cod liver oil no. 2, U.S.P., b u t the corresponding requirement was lower in the case of the other two sources of vitamin D . (3) A high incidence of crooked breastbones a t four weeks of age was observed to be associated with low levels of vitamin D . ACKNOWLEDGEMENT The authors are indebted to Dr. J. J. Waddell for supplying the irradiated animal sterol ("Delsterol") and to Dr. V. S. Asmundson for examining the data statistically. REFERENCES
Carver, J. S., and M. Rhian, 1942. The vitamin D requirement of turkey poults. Poultry Sci. 21: 260-262. Hammond, J. C, 1941. The vitamin D requirement of turkey poults. Poultry Sci. 20:204-205. Jukes, T. H„ and T. D. Sanford, 1939. The vitamin D requirements of young turkeys. J. Nutrition 18:71-85. Willgeroth, G. B., J. L. Halpin, H. R. Halloran, and J. C. Fritz. Turkeys and the vitamin D assay. Paper presented before 100th meeting, American Chemical Society, Pittsburgh, 1943.
Downloaded from http://ps.oxfordjournals.org/ at Univ of Iowa-Law Library on June 19, 2015
peared to be more efficacious for turkeys than for chicks under these conditions. I t was reported by Carver and Rhian (1942) t h a t 80 A.O.A.C. chick units of "fortified cod liver oil" per 100 grams of feed supplied adequate vitamin D for turkey poults. This does not necessarily conflict with the present observations, for if "fortified cod liver oil" contained activated animal sterol, for example, as a fortification, such an oil would tend to be more efficacious than the U.S.P. reference cod liver oil for turkeys.