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The Effectiveness of the Vitamin D Analog 1 α-OH-D3 in Promoting Fertility and Hatchability in the Laying Hen1
The Effectiveness of the Vitamin D Analog 1 a-OH-D3 in Promoting Fertility and Hatchability in the Laying Hen1 J. H. SOARES, JR., M. R. SWERDEL, and M. A. OTTINGER Department of Poultry Science, University of Maryland, College Park, Maryland 20742 (Received for publication August 31, 1978)
INTRODUCTION During the past decade considerable new data has been published to further the understanding of the role of vitamin D in calcium and phosphorus metabolism. It is clearly established that hydroxylation of vitamin D must occur in the liver and kidney for complete metabolic activity. In contrast to the great deal of effort that has been placed on the biochemistry and nutritional effectiveness of these hydroxylated metabolities, relatively little research has been published on their reproductive supporting capabilities. Manley et al. (1978) have reported that adding 1100 ICU of 25 hydroxycholecalciferol (25-OH-D3 )/kg of feed to a basal diet adequate in all nutrients including 2200 ICU vitamin D 3 /kg resulted in a significant improvement in hatchability of fertile eggs of Broad Breasted Large White turkey hens. Abdul-Rahim et al. (1977) have reported low hatchability and carry-over of vitamin D 3 activity from eggs of hens fed l,25-(OH) 2 -D 3 or 1 a-OH-D 3 . Norman et al. (1978) recently presented data of a 30 week study in which chicks were fed D3, l,25-(OH) 2 -D 3 or 24, 25-(OH) 2 -D 3 singly or in combination from 1 day of age. The group receiving the combination of l,25-(OH) 2 -D3 plus 24, 25-(OH) 2 -D 3 grew and reproduced significantly better than all other groups. In
'Scientific Article No. 2502. Contribution No. 5533 of the Maryland Agricultural Experiment Station (Department of Poultry Science).
another study, Sunde et al. (1978) reported gross reproductive failure in the form of low hatchability when laying hens were fed 1,25(OH) 2 -D 3 . As many as 80 to 90% of the hens given this metabolite produced eggs that failed to hatch. The embryos appeared to die after 18 days incubation and the only defect detected was a shortened upper mandible. The 25-OHD 3 metabolite appeared to be more effective than the dihydroxy metabolite but less effective than vitamin D 3 . Therefore, an experiment was designed to further study the effects of feeding the synthetic analogue 1 a-OH-D3 as the only source of vitamin D 3 on the reproductive performance of caged Single Comb White Leghorn hens in the final weeks of the typical 52-week production cycle. MATERIALS AND METHODS Forty Single Comb White Leghorn hens of the DeKalb strain in production for 16 weeks were placed in individual wire-floored laying cages for 20 weeks. They were fed a complete corn-soybean meal ration (Table 1) containing either 20 /ig/vitamin D 3 or 5 jug 1 a-OH-D3 /kg feed. Since this regimen was continued for 21 weeks, and egg production, egg shell quality, and calcium metabolism were also being studied, a negative control group could not be included. There was no other form of vitamin D added to the diets. The birds were housed in a temperature controlled room, fed ad libitum via individual aluminum feeders, and had free access to tap water. The room was windowless
1004
Downloaded from http://ps.oxfordjournals.org/ at New York University on April 17, 2015
ABSTRACT Single Comb White Leghorn hens were fed either 20 jug vitamin D3 or 5 Mg 1 a-OHD3 per kilogram diet for 20 weeks and then artificially inseminated. All eggs laid during the twenty-first week of feeding were collected and incubated. There were no differences in egg production or fertility between the two groups. However, the hens fed 1 a-OH-D, had significantly lower hatchability and a markedly higher incidence of embryonic abnormalities, indicative of a vitamin D deficiency. These data indicate that 1 a-OH-D3 , a synthetic analogue of the hormone form of vitamin D 3 , may be poorly transported into the egg. 1979 Poultry Science 58:1004-1006
RESEARCH NOTE TABLE 1. Basal diet fed SCWL bens g/100 g of diet
Total
70.618 19.070 2.270 6.500 .042 .900 .500 .050 .050 100.000
Provides Protein % Calcium T Phosphorus %
15.60 3.25 .71
Vitamin mix provides the following mg/kg of diet: riboflavin 1.5, calcium pantothenate 2, niacin 5, vitamin E 5, vitamin B, 2 .002, vitamin A 5.59 (325,000 IU/g), and ethoxyquin 5.65. Trace mineral mix provides the following in mg/kg diet: manganese 60; iron 25; copper 2; iodine 1.2; and zinc 18.
and, therefore, irradiation b y sunlight was minimized. T h e feed was freshly m i x e d every o t h e r m o n t h and s t o r e d in a refrigerator at 10 C t o minimize o x i d a t i o n . After 2 0 weeks of feeding t h e respective diets, each hen was artificially inseminated twice (at 1 0 0 0 and at 1400 hr t o o p t i m i z e fertility) with pooled rooster semen collected
within 30 m i n u t e s of use. All eggs p r o d u c e d b y t h e hens were collected, stored for 3 days at 10 C, and t h e n i n c u b a t e d . Egg p r o d u c t i o n , p e r c e n t fertility, p e r c e n t hatchability a n d e m b r y o n i c abnormalities were d e t e r m i n e d for each hen. An analysis of variance was performed o n t h e fertility and h a t c h a b i l i t y data t o d e t e r m i n e statistical significance (Sokal and Rohlf, 1 9 7 3 ) .
RESULTS AND DISCUSSION T h e results (Table 2) s h o w t h a t feeding t h e m e t a b o l i t e 1 a - O H - D 3 for 20 weeks, a m o r e t h a n a d e q u a t e period t o develop vitamin D deficiency, does n o t adversely affect egg p r o d u c t i o n or fertility rate b u t does result in a highly significant decrease in hatchability. During t h e 2 0 week feeding period, n o significant differences were seen in egg p r o d u c t i o n (unpublished d a t a ) . T h e lower survival rate of e m b r y o s in eggs from hens fed 1 a - O H - D 3 appears m a i n l y t o occur after day 18 of incubation and p r o b a b l y a b o u t day 20 since only a few of t h e dead e m b r y o s had n o t d r a w n t h e yolk sac into t h e a b d o m i n a l cavity. E m b r y o s were generally well developed e x c e p t t h a t one-third of t h e e m b r y o s n o t hatching h a d s h o r t e n e d u p p e r mandibles. T h e latter results were similar t o t h e findings of Sunde et al. ( 1 9 7 8 ) b u t m u c h less frequently observed, since these a u t h o r s f o u n d t h a t as m a n y as 80 t o 9 0 % of t h e dead e m b r y o s from l , 2 5 - ( O H ) 2 - D 3 fed hens exhibited this s y n d r o m e . Sunde et al. ( 1 9 7 8 ) p o i n t o u t t h a t shortened u p p e r m a n d i bles were also observed when hens were fed a vitamin D deficient diet for 3 weeks.
TABLE 2. Reproductive performance of laying bens fed either vitamin D3 or 1 u-OH-D^ la-OH-D, No. hens laying Eggs laid a No. fertile hens Average hen-day production % Fertility 0 % Hatchability 0 % Abnormalities"
19 86 16
18 80 13
.65 67.0 ± 3.65 58.4 ± 3.28 0
.64 61.7 ±6.16 45.4 C ± 5.19 33.0-
Total for the 21st week. Mean ± SEM based on the production of hens laying at least one fertile egg duringthe 21st week of the experiment. Percentage of total eggs. Significantly different from the control group (P<.05). Based on the total number of dead embryos.
Downloaded from http://ps.oxfordjournals.org/ at New York University on April 17, 2015
Ingredient Corn, yellow Soybean meal—49% Defluorinated phosphate Limestone Methionine Crude corn oil Sodium chloride (iodized) Vitamin premix a Trace mineral mix''
1005
1006
SOARES, JR. ET AL. of table eggs.
REFERENCES Abdul-Rahim, S., M. B. Patel, and J. McGinnis, 1977. Effect of vitamin D 3 and some vitamin D 3 metabolites on egg production, hatchability of eggs and vitamin D activity carry-over in chicks. Poultry Sci. 56:1691 (Abstr.). Manley, J. M., R. A. Voitle, and R. H. Harms, 1978. The influence of hatchability of turkey eggs from the addition of 25-hydroxycholecalciferol to the diet. Poultry Sci. 57:290-292. Norman, A. W., T. Okamoto, and H. Henry, 1978. Differential response of male and female chicken to dihydroxylated vitamin D metabolites. Fed. Proc. 37:409 (Abstr.). Sokal, R. R., and F. J. Rohlf, 1973. Assumptions of analysis of variance. Page 210—217 in Introduction to biostatistics. D. Kennedy and R. B. Park, ed. W. H. Freeman and Co., San Francisco, CA. Sunde, M. L., C. M. Turk, and H. F. DeLuca, 1978. The essentiality of vitamin D metabolites for embryonic chick development. Science 200: 1067-1069.
Downloaded from http://ps.oxfordjournals.org/ at New York University on April 17, 2015
The results reported herein for 1 a-OH-D 3 are not as severe as those reported by Sunde et al. (1978) for l,25-(OH) 2 -D 3 and indicate either a difference in the ability of the hen to transport 1 a-OH-D 3 into the egg or insufficient data. In any case, it appears likely that neither l,25-(OH) 2 -D 3 nor 1 a-OH-D 3 have adequate accessability to the egg and therefore can not presently be recommended for potential use in breeding diets. However, the fact that 1 a-OHD 3 and l,25-(OH) 2 -D 3 (the latter satisfies the definition of a hormone) do not appear to be readily transferred into the egg may well be a benefit in the regulation of production of table eggs if either is ever used in diets of hens reared for such production. This would result in very little consumption by human beings of the hormone form of vitamin D 3 . Perhaps the ultimate future use of these metabolites in egg production will be to combine one or the other with vitamin D 3 or 25-OH-D3 at specific dose levels to optimize egg shell and nutrient quality
INTRODUCTION During the past decade considerable new data has been published to further the understanding of the role of vitamin D in calcium and phosphorus metabolism. It is clearly established that hydroxylation of vitamin D must occur in the liver and kidney for complete metabolic activity. In contrast to the great deal of effort that has been placed on the biochemistry and nutritional effectiveness of these hydroxylated metabolities, relatively little research has been published on their reproductive supporting capabilities. Manley et al. (1978) have reported that adding 1100 ICU of 25 hydroxycholecalciferol (25-OH-D3 )/kg of feed to a basal diet adequate in all nutrients including 2200 ICU vitamin D 3 /kg resulted in a significant improvement in hatchability of fertile eggs of Broad Breasted Large White turkey hens. Abdul-Rahim et al. (1977) have reported low hatchability and carry-over of vitamin D 3 activity from eggs of hens fed l,25-(OH) 2 -D 3 or 1 a-OH-D 3 . Norman et al. (1978) recently presented data of a 30 week study in which chicks were fed D3, l,25-(OH) 2 -D 3 or 24, 25-(OH) 2 -D 3 singly or in combination from 1 day of age. The group receiving the combination of l,25-(OH) 2 -D3 plus 24, 25-(OH) 2 -D 3 grew and reproduced significantly better than all other groups. In
'Scientific Article No. 2502. Contribution No. 5533 of the Maryland Agricultural Experiment Station (Department of Poultry Science).
another study, Sunde et al. (1978) reported gross reproductive failure in the form of low hatchability when laying hens were fed 1,25(OH) 2 -D 3 . As many as 80 to 90% of the hens given this metabolite produced eggs that failed to hatch. The embryos appeared to die after 18 days incubation and the only defect detected was a shortened upper mandible. The 25-OHD 3 metabolite appeared to be more effective than the dihydroxy metabolite but less effective than vitamin D 3 . Therefore, an experiment was designed to further study the effects of feeding the synthetic analogue 1 a-OH-D3 as the only source of vitamin D 3 on the reproductive performance of caged Single Comb White Leghorn hens in the final weeks of the typical 52-week production cycle. MATERIALS AND METHODS Forty Single Comb White Leghorn hens of the DeKalb strain in production for 16 weeks were placed in individual wire-floored laying cages for 20 weeks. They were fed a complete corn-soybean meal ration (Table 1) containing either 20 /ig/vitamin D 3 or 5 jug 1 a-OH-D3 /kg feed. Since this regimen was continued for 21 weeks, and egg production, egg shell quality, and calcium metabolism were also being studied, a negative control group could not be included. There was no other form of vitamin D added to the diets. The birds were housed in a temperature controlled room, fed ad libitum via individual aluminum feeders, and had free access to tap water. The room was windowless
1004
Downloaded from http://ps.oxfordjournals.org/ at New York University on April 17, 2015
ABSTRACT Single Comb White Leghorn hens were fed either 20 jug vitamin D3 or 5 Mg 1 a-OHD3 per kilogram diet for 20 weeks and then artificially inseminated. All eggs laid during the twenty-first week of feeding were collected and incubated. There were no differences in egg production or fertility between the two groups. However, the hens fed 1 a-OH-D, had significantly lower hatchability and a markedly higher incidence of embryonic abnormalities, indicative of a vitamin D deficiency. These data indicate that 1 a-OH-D3 , a synthetic analogue of the hormone form of vitamin D 3 , may be poorly transported into the egg. 1979 Poultry Science 58:1004-1006
RESEARCH NOTE TABLE 1. Basal diet fed SCWL bens g/100 g of diet
Total
70.618 19.070 2.270 6.500 .042 .900 .500 .050 .050 100.000
Provides Protein % Calcium T Phosphorus %
15.60 3.25 .71
Vitamin mix provides the following mg/kg of diet: riboflavin 1.5, calcium pantothenate 2, niacin 5, vitamin E 5, vitamin B, 2 .002, vitamin A 5.59 (325,000 IU/g), and ethoxyquin 5.65. Trace mineral mix provides the following in mg/kg diet: manganese 60; iron 25; copper 2; iodine 1.2; and zinc 18.
and, therefore, irradiation b y sunlight was minimized. T h e feed was freshly m i x e d every o t h e r m o n t h and s t o r e d in a refrigerator at 10 C t o minimize o x i d a t i o n . After 2 0 weeks of feeding t h e respective diets, each hen was artificially inseminated twice (at 1 0 0 0 and at 1400 hr t o o p t i m i z e fertility) with pooled rooster semen collected
within 30 m i n u t e s of use. All eggs p r o d u c e d b y t h e hens were collected, stored for 3 days at 10 C, and t h e n i n c u b a t e d . Egg p r o d u c t i o n , p e r c e n t fertility, p e r c e n t hatchability a n d e m b r y o n i c abnormalities were d e t e r m i n e d for each hen. An analysis of variance was performed o n t h e fertility and h a t c h a b i l i t y data t o d e t e r m i n e statistical significance (Sokal and Rohlf, 1 9 7 3 ) .
RESULTS AND DISCUSSION T h e results (Table 2) s h o w t h a t feeding t h e m e t a b o l i t e 1 a - O H - D 3 for 20 weeks, a m o r e t h a n a d e q u a t e period t o develop vitamin D deficiency, does n o t adversely affect egg p r o d u c t i o n or fertility rate b u t does result in a highly significant decrease in hatchability. During t h e 2 0 week feeding period, n o significant differences were seen in egg p r o d u c t i o n (unpublished d a t a ) . T h e lower survival rate of e m b r y o s in eggs from hens fed 1 a - O H - D 3 appears m a i n l y t o occur after day 18 of incubation and p r o b a b l y a b o u t day 20 since only a few of t h e dead e m b r y o s had n o t d r a w n t h e yolk sac into t h e a b d o m i n a l cavity. E m b r y o s were generally well developed e x c e p t t h a t one-third of t h e e m b r y o s n o t hatching h a d s h o r t e n e d u p p e r mandibles. T h e latter results were similar t o t h e findings of Sunde et al. ( 1 9 7 8 ) b u t m u c h less frequently observed, since these a u t h o r s f o u n d t h a t as m a n y as 80 t o 9 0 % of t h e dead e m b r y o s from l , 2 5 - ( O H ) 2 - D 3 fed hens exhibited this s y n d r o m e . Sunde et al. ( 1 9 7 8 ) p o i n t o u t t h a t shortened u p p e r m a n d i bles were also observed when hens were fed a vitamin D deficient diet for 3 weeks.
TABLE 2. Reproductive performance of laying bens fed either vitamin D3 or 1 u-OH-D^ la-OH-D, No. hens laying Eggs laid a No. fertile hens Average hen-day production % Fertility 0 % Hatchability 0 % Abnormalities"
19 86 16
18 80 13
.65 67.0 ± 3.65 58.4 ± 3.28 0
.64 61.7 ±6.16 45.4 C ± 5.19 33.0-
Total for the 21st week. Mean ± SEM based on the production of hens laying at least one fertile egg duringthe 21st week of the experiment. Percentage of total eggs. Significantly different from the control group (P<.05). Based on the total number of dead embryos.
Downloaded from http://ps.oxfordjournals.org/ at New York University on April 17, 2015
Ingredient Corn, yellow Soybean meal—49% Defluorinated phosphate Limestone Methionine Crude corn oil Sodium chloride (iodized) Vitamin premix a Trace mineral mix''
1005
1006
SOARES, JR. ET AL. of table eggs.
REFERENCES Abdul-Rahim, S., M. B. Patel, and J. McGinnis, 1977. Effect of vitamin D 3 and some vitamin D 3 metabolites on egg production, hatchability of eggs and vitamin D activity carry-over in chicks. Poultry Sci. 56:1691 (Abstr.). Manley, J. M., R. A. Voitle, and R. H. Harms, 1978. The influence of hatchability of turkey eggs from the addition of 25-hydroxycholecalciferol to the diet. Poultry Sci. 57:290-292. Norman, A. W., T. Okamoto, and H. Henry, 1978. Differential response of male and female chicken to dihydroxylated vitamin D metabolites. Fed. Proc. 37:409 (Abstr.). Sokal, R. R., and F. J. Rohlf, 1973. Assumptions of analysis of variance. Page 210—217 in Introduction to biostatistics. D. Kennedy and R. B. Park, ed. W. H. Freeman and Co., San Francisco, CA. Sunde, M. L., C. M. Turk, and H. F. DeLuca, 1978. The essentiality of vitamin D metabolites for embryonic chick development. Science 200: 1067-1069.
Downloaded from http://ps.oxfordjournals.org/ at New York University on April 17, 2015
The results reported herein for 1 a-OH-D 3 are not as severe as those reported by Sunde et al. (1978) for l,25-(OH) 2 -D 3 and indicate either a difference in the ability of the hen to transport 1 a-OH-D 3 into the egg or insufficient data. In any case, it appears likely that neither l,25-(OH) 2 -D 3 nor 1 a-OH-D 3 have adequate accessability to the egg and therefore can not presently be recommended for potential use in breeding diets. However, the fact that 1 a-OHD 3 and l,25-(OH) 2 -D 3 (the latter satisfies the definition of a hormone) do not appear to be readily transferred into the egg may well be a benefit in the regulation of production of table eggs if either is ever used in diets of hens reared for such production. This would result in very little consumption by human beings of the hormone form of vitamin D 3 . Perhaps the ultimate future use of these metabolites in egg production will be to combine one or the other with vitamin D 3 or 25-OH-D3 at specific dose levels to optimize egg shell and nutrient quality