- Email: [email protected]
25-Hydroxycholecalciferol Fails to Prevent Tibial Dyschondroplasia in Broiler Chicks Raised in Battery Brooders
01999Applied Poultly Science, Inc
25-HYDROXYCHOLECALCIFEROL FAILS TO PREVENT TIBIAL DYSCHONDROPLASIA IN BROILER CHICKS RAISED IN
BATTERY BROODERS
Primary Audience: Broiler Producers, Extension Specialists, Nutritionists, Researchers
TD-inducing diet [l] or a diet with a normal DESCRIPTION OF PROBLEMcalcium:phosphorus ratio [6]. Five ,u@g was The effectiveness of the vitamin D3 metabolite, 1,25-dihydroxycholecalciferol (1,25-(OH)ZD3) in preventing tibial dyschondroplasia (TD) in broiler chickens has been clearly documented for birds grown both in wire-floored cages and on litter [1,2,3, 4, 51. This metabolite is also known to be effective when added to either a low-calcium
sufficient when fed to chicks in batterybrooder cages [2] and 6 pgkg minimized the incidence and severityof TD in broilers raised on wood shavings [5]. Cell differentiation of chicken chondrocytes is known to be stimulated by which therefore prevents 1,25-(OH),D3 [?‘I, the accumulation of prehypertrophic cells
1 Present address: Dept. of Animal Science, Michigan State University, E. Lansing, MI
48824.
Downloaded from http://japr.oxfordjournals.org/ at North Dakota State University on June 19, 2015
KEVIN D. ROBERSON~ Department of Animal Science, University of Tennessee, Martin Bperiment Station, Martin, TN 38238 Phone: (901) 587-7279 F M : (901) 587-7968
Research Report ROBERSON
on growth performance and the incidence and severity of TD in commercial broiler chicks raised in battery-brooder cages and fed normal or low calcium levels. The effect of 25-(OH)D3 on phytate phosphorus utilization was also investigated. The replacement of cholecalciferol with 25-(OH)D3 has been reported to decrease the phosphorus requirement of broilers [14, 151 and 1,25-(OH)ZD3 is known to increase phytate phosphorus utilization in chickens [16].
MATERIALS AND METHODS A total of 192 Ross X Ross chicks were donated by a local hatchery [17] for Experiment 1.The hatching eggs were vaccinated for Marek's Disease prior to placement in the hatcher. The chicks were spray{accinated for infectious bronchitis and Vewcastle disease on the day of hatch. Dayild chicks were sexed and males and females were evenly distributed in each pen of an elec.rically heated battery-brooder with raised wire floors and incandescent bulbs (181. In Zxperiment 1,25-(OH)D3was added at 0,23, l6,69, or 92pgkg to a control corn-soybean neal diet containing 0.95% calcium (Table 1).
INGREDIENT
m-
CONTROL
INDUCING 70
%
Ground yellow corn
53.65
5651
Soybean meal (48%)
36.69
Choice white erease
5.14
35.00 5.00
Dicalcium phosphate
1.57
Limestone
1.33
I
0.28
0.45
1.86
I Salt
I
0.45
I
I nr .methinnine
I
n?n
I
.
.L"L.I.I.
,".I.
II rhrnmiv nvirl..,ILLY.....% "*I.,-
I
*"."I
".W
".W
nin
I
I
Irnt.i I
I
1
".I"
innnn A " " . " "
I I I
I I
n in " . I " ~ M M I " " . " "
I * IIaLcb L l l l L l C l a l yllrllllA p " " r u G u
111 11.
"(j "I
UIS,. &,11L,
100; iron, 60; manganese, 120; copper,& selenium,O.l; iodine, 2.1.
%itamin premc provided in mg/kg ditt. (except >\. . .acetate ~of ~ -.. . 5s noreu): vlrarnin A,JUW IU trerinyi wramin E, 11 IU (dla-toco hero1 acetate); vitamin I$,309 ICU; menadione bisulgte, 1.1; thiamine, 2.2; riboflavin, 4.4; ~
.I.
-~.&.
L-nnnITI,
chloride, 600; ethoxyquin, 125.
Downloaded from http://japr.oxfordjournals.org/ at North Dakota State University on June 19, 2015
in the growth plate that is characteristic of a dyschondroplastic lesion. Any vitamin D metabolite hydroxylated at the 1-C position can effectively decrease TD in broilers [2]. However, 25-hydroxycholecalciferol (Z-(OH)D3) was not effective when fed at lOpg/kg in addition to a diet adequate (27.5pg/kg) in cholecalciferol [l]. The vitamin D3 receptor is known to discriminate against 25-(OH)D3 in favor of 1,25-(OH)2@ [gl. Recently, interest has increased in the use of 25-(OH)D3 at higher levels than previously investigated to alleviate TD in broilers. It has recently been reported that the replacement of cholecalciferol with 69 pgkg of Z-(OH)D3 would decrease TD in broilers raised in floor pens [9]. However, similar results were not observed in battery-brooder studies.This level of 25-(OH)D3 was chosen after Yarger et al. [lo] reported that 69 pgkg in place of cholecalciferol would increase body weight and improve feed efficiency in market age broilers. Mireles et al. [9] reported increased body welght and feed intake in one experiment, but decreases in these parameters in a later experiment. Rennie and Whitehead [ l l ] found that replacement of 75 p g k g cholecalciferol with 25-(OH)D3decreased TD in one of two experiments when chicks were raised in batterybrooder cages. f i e addition of 250 pgkg 25-(OH)D3 almost completely eliminated the incidence of TD in broilers. In contrast to the report by Mireles ef al. [9], there was no effect of 25-(OH)D3 on the incidence of TD in birds raised on litter. The incidence of TD was low in this experiment, but severity was still decreased by B-(OH)D3. Genetics reportedly has a role in the effectiveness of 25-(OH)D3 and other vitamin D metabolites in preventing TD in broilers [12]. When added to a calcium-adequate diet containing 27.5 pg/kg cholecalciferol, 5 puglkg 25-(OH)D3 reduced TD and 10 pgkg maximized bone ash in chicks selected for a low incidence of TD. Zhang et al. [13] also reported a decrease in the incidence of TD in broiler chickens selected for a low incidence of TD when Z-(OH)D3 was fed, but no response was observed in chickens bred for a high incidence of TD. The objective of this study was to determine the effects of graded levels of 25-(OH)D3
55
56
rinized samples for measurement of phosphorus [24] and calcium [25] by colorimetric procedures. Excreta samples were collected from each pen in each experiment and saved for measurement of chromic oxide [26] and phytate phosphorus [27 in order to calculate phytate phosphorus retention values [B]. The experiments were terminated when the birds reached 17 days of age. All chicks were killed by cervical dislocation after body weight determination. The right tibia of each chick was scored for incidence and severity of TD by making a longitudinal cut across the proximal end of the bone [23]. The severity scores of TD ranged from 0 (no lesion) to 3 (severe lesion). The left tibia was saved for bone ash analysis [29]. The data were analyzed as described in previous publications [30,31, 32,331. In Experiment 1,both treatments containing 0 pgkg 25-(OH)D3 were analyzed as one treatment since there was adequate cholecalciferol in each treatment and there were no differences in any parameter measured.
RESULTS AND DISCUSSION There was no effect on body weight in either experiment (Tables 2 and 3). This agrees with the results of Edwards [l].However, Yarger et al. [lo] reported that 69 pgkg 25-(OH)D3 in place of a similar amount of cholecalciferol increased body weight of chickens grown to about 50 days of age in nine of 10 floor pen experiments. There was no difference in body weight in the one experiment in which 25-(OH)D3 was not effective. Rennie and Whitehead [11] did not observe a difference in body weight of 3-wk-old chicks grown in wire cages. However, they observed a decrease in 3- and 6-wk-old chickens fed 75pg/kg 25-(OH)D3 in place of cholecalciferol. Mireles et al. [9] found inconsistent results of 25-(OH)D3 on body weight of broilers grown to 7 wk of age on litter. Body weight was increased in one experiment, but decreased in another when 69 pgkg was fed. Feed efficiency was also not affected by 25-(OH)D3 in these studies. Yarger et al. [lo] reported better feed efficiency with 69 pgkg Z-(OH)D3. However, no feed efficiency responses were found by others [1, 11,121. There were no effects of 25-(OH)D3 on plasma calcium or phosphorus (Table 3). Rennie and Whitehead [ll] did not observe significant effects on plasma calcium and
Downloaded from http://japr.oxfordjournals.org/ at North Dakota State University on June 19, 2015
Crystalline 25-(OH)D3 was donated [19] and diluted to 140 mgkg with rice hulls. This premix was used to make the appropriate additions to the experimental diets. Another treatment consisted of supplemental cholecalciferol at 69p& to see if additional cholecalciferol yielded any response to 25-(OH)D3. Crystalline cholecalciferol [20] was diluted with rice hulls to 140 mgkg also. The experimental diets were fed to four pens of eight chicks per treatment. The horizontal mixer used to mix the diets was cleaned and sand was run through the mixer and auger that delivered the feed after mixing. Although cholecalciferol was not added to the premix, the complete basal diet was analyzed [21] to contain adequate ( > 20,ugkg) vitamin D activity [Z]. The mixer was also being used at this time to mix experimental swine diets containing a very high (25Opgkg added) level of cholecalciferol. Chromic oxide was added at 0.1% as an external indicator to determine phytate phosphorus retention values. The second experiment utilized 240 straight run chicks from the same source as Experiment 1. In Experiment 2, four treatments were allocated to six pens of 10 chicks each fed a low-calcium, TD-inducing diet as described by Edwards and Veltmann [23]. Crystalline cholecalciferol or 25-(OH)D3 was diluted in propylene glycol for liquid addition to the feed to see if the form of 25-(OH)D3 affected the results by supplying a more consistent source under the conditions of the study. The treatments were as follows: 1) 7 0 p g k g cholecalciferol, 2) 70 pgkg 25-(OH)D3, 3) 250 p g k g 25-(OH)D3, and 4) 250pg/kg 25-(OH)D3 added in dry form using the premix described previously. Analysis of the control diet prior to cholecalciferol supplementation indicated that vitamin D activity was present in the mixed feed at approximately 22pg (880 ICU)/kg. The brooder was placed in a room located at the dairy unit of the Martin Experiment Station, University of Tennessee, Martin, TN. Dark plastic was placed over all outside windows in the room. Clear plastic was placed over fluorescent lights (no diffusers) located on the ceiling. Lighting was continuous in the room and the battery-brooder. At 16 days of age, blood samples were obtained by cardiac puncture from one chick per pen. Plasma was extracted from the hepa-
25-HYDROXYCHOLECALCIFEROL
Research Report
57
ROBERSON
Downloaded from http://japr.oxfordjournals.org/ at North Dakota State University on June 19, 2015
25-HYDROXYCHOLECALCIFEROL 58
JAPR
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(j
Research Report 59
ROBERSON
even when TD incidence is low. However, considering that higher levels of 25-(OH)D3 used in both experiments in this study did not influence the severity of TD in the chicks, it cannot be concluded from this study that 25-(OH)D3 can effectivelydecrease the severity of TD. There was no consistent effect of 25(OH)D3 on phytate phosphorus utilization in these experiments. In Experiment 2, there was a decrease (P < .OS). in phytate phosphorus retention when 25-(OH)D3 was fed, but no effect was observed in Experiment 1.Previous unpublished work in this laboratory found no effect of dietary E-(OH)D3 on phytate phosphorus utilization. Although 1,25(OH)2D3has been shown to increase phytate phosphorus retention [13, 34, 351, the author is not aware of any reports on the effect of E-(OH)D3 on this parameter. Rennie and Whitehead [ l l ] hypothesized that high levels of 25-(OH)D3 may affect chondrocyte differentiation by binding to the vitamin D receptor and activating Dresponsive genes or by inducing endogenous synthesis of l-hydroxylated metabolites. Yarger et al. [36] did not observe a change in serum 1,25-(OH)zD3 when 25-(OH)D3 was fed at 207 pgkg. However, there appears to be no relationship between blood levels of 1,25-(OH)@3 and the incidence of TD in broilers [5,12,34,37l. Studies on the relationship between genetics and nutrition [12, 131 suggest that when broiler chickens are bred for a low incidence of TD, additives such as 25-(OH)D3 may be effective because of their interaction with other causes of the cartilage abnormality- causes related to nutrition or to some factor other than genetics. Although researchers report that dietary 25-(OH)D3 may alleviates TD in broilers, there was no indication in these studies of any effect in chicks raised in cages.
CONCLUSIONS AND APPLICATIONS 1. Supplementation of 25-(OH)D3 at levels as high as 250pglkg did not prevent TD or affect growth performance in commercial broiler chicks raised in a battery-brooder. 2. Level of dietary calcium and form of 25-(OH)D3 addition did not affect the response observed with supplementation of 25-(OH)D3.
Downloaded from http://japr.oxfordjournals.org/ at North Dakota State University on June 19, 2015
phosphorus when high levels of 25-(OH)D3 alone were added to the diet. There was no indication that 25-(OH)D3 was able to reduce the incidence of TD in broiler chicks regardless of dietary calcium level or form of Z-(OH)D3 addition (Tables 2 and 3). This agrees with the data of Edwards [l]and one of two experiments reported by Rennie and Whitehead [ll]. In the experiment in which Rennie and Whitehead [ll]did not see a response, the incidence of TD in the chicks fed cholecalciferolwas 28%. The incidence of TD in the control diet was 34% in Experiment 1and 24% in Experiment 2 in this study. When 25-(OH)D3 was found to be effective by Rennie and Whitehead [ll], the incidence of TD in chicks fed cholecalciferol was 64%. Mitchell and Edwards [12] observed a decrease in TD incidence and severity when 25-(OH)D3 was added to a calcium-adequate (0.95 %) and cholecalciferol- a de quat e (27.5pgkg) diet fed to chicks bred for a low incidence of TD. The incidence of TD in this line when Z-(OH)D3 was not added was 44 and 61%, respectively, in the two experiments in which25-(OH)D3 was studied. Hence, from their studies, it appears that a high incidence of TD must be present to elicit a response from dietary25-(OH)D3.However, Mireleset al. [9] reported a significant decrease in TD incidence when lowered from 30 to 13%. Mireles etal. [9] utilized 500 birds per treatment, which is much higher than the number of chicks used in previous studies [ll,121or the present experiments. The severity of TD was generally not affected by 25-(OH)D3 (Tables 2 and 3). However, there was a significant decrease in the severity of TD when 46 pg/kg 25-(OH)D3 was fed compared to the diets in which no 25-(OH)D3 was fed in Experiment 1.Rennie and Whitehead [ll]found that 75 pgkg Z-(OH)D3 could decrease TD in broilers
JAPR 25-HYDROXYCHOLECALCIFEROL
60
REFERENCES AND NOTES 1. Edwards, H.M., Jr., 1989. The effect of dietary cholecalciferol, 25-hydroxycholecalciferol and 1,25dihydroxycholecalciferol on the development of tibial dyschondroplasia in broiler chickens in the absence or presence of disulfiram. J. Nutr. 119647-652. 2. Edwards, H.M., Jr., 1990. Efficacy of several vitamin D compounds in the prevention of tibial dyschondroplasia in broiler chickens. J. Nutr. 12010541061.
4. Rennie, J.S., C.C. Whitehead, and B.H. Thorp, 1993. The effect of dietary 1,25dihydroxycholecalciferol in preventing tibial dyschondroplasia in broilers fed on diets imbalanced in calcium and phosphorus. Br. J. Nutr. 69:809-816.
5. Roberson, K.D. and EM.Edwards,Jr., 1996. Effect of dietary 1,2.5-dihydroxycholecalciferollevel on broiler performance. Poultry Sci. 7590-94. 6. Rennie, J.S., H A McCormaclc, C. Farquharson, J.L Berry, EB. Mmer, and C.C. Whitehead, 1995.Interaction between dieta 1,25dihydroxycholecalciferol and calcium and effects ormanagement on the occurrence of tibial dyschondroplasia, le abnormalities, and performance in broiler chickens. br. Poultry Sci. 36:4654?7. 7. Fuqnhprson, C., kS.Law, E Seawright, D.W. Burt, and C.C. Whitehead, 1993. In y i y ~effect of 1,25dihydroxycholecalciferol on the proliferation and differentiation of avian chondrocytes. J. Bone and Min. Res. 81081-1088. 8. Hughes, M.R, D.J. Baylink, P.G. Jones, and M.R Haossler, 1976. Radioligand receptor assay for 25hydraxyvitamin Dm3. Application to hypervitaminosis D J . Clin. Invest. 5861-70.
9. Mireles, A, Jr., S. Kim, B. Krautmann, J. Yarger, and L Siark, 1996. Effect of 25-hydroxycholecalciferol (z-(OH)D3) on broiler field performance and incidence of tibial dyschondro lasia(TD): Minimum D3 metabolite consumption periodq Poultry Sci. 75(Suppl):280 (Abs). 10. Yarger, J.G., CA. Saunders, J.L McNaughton, C.L Quarles, B.W.Hollis, and RW. Gray, 1995. Comparison of dietary25-hydro holecalciferol and cholecalciferol in broiler chickens. %tty Sci. 741159-1167. 11.Rennie, J.S. and C.C. Whitehead,1996. Effectiveness of dietary 25- and 1-hydroxycholecalciferolin combating tibial dyschondroplasia in broiler chickens. Br. Poultry Sci. 37413-421. 12. Mitchell, RD. and H.M. Edwards,Jr., 1997. The effects of ultraviolet light and cholecalciferol and its metabolites on the development of leg abnormalities in chickens genetically selected for a high or low incidence of tibial dyschondroplasia. Poultry Sci. 76346354. 13. Zhang, X,G. Uu,G.R McDaniel, and D.A. Roland, 1997. Responses of broiler lines selected for tibial dyschondroplasia incidence t o s u p lementary 25hydroxycholecalciferol.J. Appl. Poultrykes. 6410-416. 14. McNaoghton, J.L, EJ. Day, and B.C. Dilworlh, 1977. T h e chick's requirement for 25-hydroxy-
15. McNaughton, J.L and R Murray, 1990. Effect of 2.5-hydroxycholecalciferol (25-(OH)D!) and vitamin D3 on hos horus requirement of broilers. Poultry Sci. 69({ uppl .178 (Abs).
7.
16. Edwards, H.M., Jr., 1993. Dietary 1,25-dihydroxycholecalciferol supplementation increases h ate phosphorus utilization in chickens. J. Nutr. 123!6&7. 17. Seaboard Farms, Mayfield, KY 42066. 18. Petersime Incubator Co., Gettysburg, OH 45328.
19. Amoco BioProducts Corporation, Napenille, IL 60563. 20. Amersham, Arlington Heights, 1L 6ooo5.
21. Woodson-Tenent Laboratories, Inc., Memphis, TN 38101. 2 2 Edwards,H.M., Jr., M.A. Elliot, S. Sooncharernyin& and W.M. Brillon, 1994. Quantitative requirement
for cholecalciferol in the absence of ultraviolet light. Poultry Sci. 73288-294. 23. Edwards, H.M., Jr. and J . R Vellmann, 1983. The role of calcium and phosphorus in the etiology of tibial dyschondroplasia in young chicks. J. Nutr. 1131568-1575. 24. Goldenberg, H. and A. Fernandeq 1966. Simplified method for the estimation of inorganic phosphorus in body fluids. Clin. Chem. 128714382.
25. Moorehead, W.R and H.G. B b 1974.2-amino2-methyl-1-propanol as the alkalizing agent in an improved continuous flow cresolphtalein com lexone procedure for calcium in serum. Clin. Chem. &:14581460. 26. Fenlon, T.W. and M. Fenton, 1979. An improved procedure for the determination of chromic oxide in feed and feces. Canadian J. Anim. Sci. 59:631-634. 27. Common, RH., 1940. The phytic acid content of some poultry feeding stuffs. Analyst 65:79-83. 28. Edwards,H.M., Jr. and M.B. Gillis, 1959. A chromic oxide balance method for determining phosphate availability. PoultIy Sci. 38569-574. 29. Associalion of Ol'f'icial Analytical Chemists, 1995. Official Methods of Analysis. 16th Edition. Assn. Offic. Anal. Chemists, Washington, DC. 30. The data were anal ed by ANOVA using the General Linear Models (GrM) rocedure of SAS [31]. Regression analysis of the 25-(8H)D3 level effect was evaluated. Treatment means were separated by Duncan's multiple range test [32] with a 5% level of probability, Orthogonal contrasts were conducted when more powerful statistical comparisons of treatment effects were desired [33].
31. SAS Institute, 1985. SAS User's Guide: Statistics. Version 5 Edition. SAS Institute, Inc., Cary, NC.
32. Duncan, D.B., 1955. Multiple range and multiple
F tests. Biometrics 11:142. 33. Sleel, RG.D. and J.H. Torrie, 1980. Princi les and Procedures of Statistics. 2nd Edition. McGraw-dl1 Book Co.,New York,NY.
Downloaded from http://japr.oxfordjournals.org/ at North Dakota State University on June 19, 2015
3. Edwards, H.M., Jr., M.A. Ellfol, a n d S. Sooncharernyhg, 1992. Effect of dietary calcium on tibial dyschondro lasia. Interaction with light, cholecalciferol, 1,25-di~ydroxycholecalciferol, protein, and synthetic zeolite. Poultry Sci. 71:2041-2055.
cholecalciferol and cholecalciferol. Poultry Sci. 56511516.
Research Report ROBERSON
61 36. Yarger, J.G., C.L Quarles, B.W. Hollis, andRW. Gray, 1995. Safety of 25-hydroxycholecalciferol as a source Of cholecalciferol in pOukry rations. Poultry SCi. 74~1437-1446.
34. Robcrson, K.D. and H.M. Edwards, Jr., 1994. Effects of ascorbic acid and 1,Zdihydroxycholecalciferol on alkaline phosphatase and tibial dyschondroplasia in broiler chickens. Br. Poultry Sci. 35:763-773.
ditive effectsof 1,25dihydro~cholecalciferoland phytase on phytate phosphorus utilization and related parameters in broiler carcasses. Poultry Sci. 75:111-119.
I
and 1,Z-dihydroxycholecalciferolon the develGment of tibial dyschondroplasia in broilers during the starter and grower periods. Poultry Sci. 74:1495-1505.
Downloaded from http://japr.oxfordjournals.org/ at North Dakota State University on June 19, 2015
25-HYDROXYCHOLECALCIFEROL FAILS TO PREVENT TIBIAL DYSCHONDROPLASIA IN BROILER CHICKS RAISED IN
BATTERY BROODERS
Primary Audience: Broiler Producers, Extension Specialists, Nutritionists, Researchers
TD-inducing diet [l] or a diet with a normal DESCRIPTION OF PROBLEMcalcium:phosphorus ratio [6]. Five ,u@g was The effectiveness of the vitamin D3 metabolite, 1,25-dihydroxycholecalciferol (1,25-(OH)ZD3) in preventing tibial dyschondroplasia (TD) in broiler chickens has been clearly documented for birds grown both in wire-floored cages and on litter [1,2,3, 4, 51. This metabolite is also known to be effective when added to either a low-calcium
sufficient when fed to chicks in batterybrooder cages [2] and 6 pgkg minimized the incidence and severityof TD in broilers raised on wood shavings [5]. Cell differentiation of chicken chondrocytes is known to be stimulated by which therefore prevents 1,25-(OH),D3 [?‘I, the accumulation of prehypertrophic cells
1 Present address: Dept. of Animal Science, Michigan State University, E. Lansing, MI
48824.
Downloaded from http://japr.oxfordjournals.org/ at North Dakota State University on June 19, 2015
KEVIN D. ROBERSON~ Department of Animal Science, University of Tennessee, Martin Bperiment Station, Martin, TN 38238 Phone: (901) 587-7279 F M : (901) 587-7968
Research Report ROBERSON
on growth performance and the incidence and severity of TD in commercial broiler chicks raised in battery-brooder cages and fed normal or low calcium levels. The effect of 25-(OH)D3 on phytate phosphorus utilization was also investigated. The replacement of cholecalciferol with 25-(OH)D3 has been reported to decrease the phosphorus requirement of broilers [14, 151 and 1,25-(OH)ZD3 is known to increase phytate phosphorus utilization in chickens [16].
MATERIALS AND METHODS A total of 192 Ross X Ross chicks were donated by a local hatchery [17] for Experiment 1.The hatching eggs were vaccinated for Marek's Disease prior to placement in the hatcher. The chicks were spray{accinated for infectious bronchitis and Vewcastle disease on the day of hatch. Dayild chicks were sexed and males and females were evenly distributed in each pen of an elec.rically heated battery-brooder with raised wire floors and incandescent bulbs (181. In Zxperiment 1,25-(OH)D3was added at 0,23, l6,69, or 92pgkg to a control corn-soybean neal diet containing 0.95% calcium (Table 1).
INGREDIENT
m-
CONTROL
INDUCING 70
%
Ground yellow corn
53.65
5651
Soybean meal (48%)
36.69
Choice white erease
5.14
35.00 5.00
Dicalcium phosphate
1.57
Limestone
1.33
I
0.28
0.45
1.86
I Salt
I
0.45
I
I nr .methinnine
I
n?n
I
.
.L"L.I.I.
,".I.
II rhrnmiv nvirl..,ILLY.....% "*I.,-
I
*"."I
".W
".W
nin
I
I
Irnt.i I
I
1
".I"
innnn A " " . " "
I I I
I I
n in " . I " ~ M M I " " . " "
I * IIaLcb L l l l L l C l a l yllrllllA p " " r u G u
111 11.
"(j "I
UIS,. &,11L,
100; iron, 60; manganese, 120; copper,& selenium,O.l; iodine, 2.1.
%itamin premc provided in mg/kg ditt. (except >\. . .acetate ~of ~ -.. . 5s noreu): vlrarnin A,JUW IU trerinyi wramin E, 11 IU (dla-toco hero1 acetate); vitamin I$,309 ICU; menadione bisulgte, 1.1; thiamine, 2.2; riboflavin, 4.4; ~
.I.
-~.&.
L-nnnITI,
chloride, 600; ethoxyquin, 125.
Downloaded from http://japr.oxfordjournals.org/ at North Dakota State University on June 19, 2015
in the growth plate that is characteristic of a dyschondroplastic lesion. Any vitamin D metabolite hydroxylated at the 1-C position can effectively decrease TD in broilers [2]. However, 25-hydroxycholecalciferol (Z-(OH)D3) was not effective when fed at lOpg/kg in addition to a diet adequate (27.5pg/kg) in cholecalciferol [l]. The vitamin D3 receptor is known to discriminate against 25-(OH)D3 in favor of 1,25-(OH)2@ [gl. Recently, interest has increased in the use of 25-(OH)D3 at higher levels than previously investigated to alleviate TD in broilers. It has recently been reported that the replacement of cholecalciferol with 69 pgkg of Z-(OH)D3 would decrease TD in broilers raised in floor pens [9]. However, similar results were not observed in battery-brooder studies.This level of 25-(OH)D3 was chosen after Yarger et al. [lo] reported that 69 pgkg in place of cholecalciferol would increase body weight and improve feed efficiency in market age broilers. Mireles et al. [9] reported increased body welght and feed intake in one experiment, but decreases in these parameters in a later experiment. Rennie and Whitehead [ l l ] found that replacement of 75 p g k g cholecalciferol with 25-(OH)D3decreased TD in one of two experiments when chicks were raised in batterybrooder cages. f i e addition of 250 pgkg 25-(OH)D3 almost completely eliminated the incidence of TD in broilers. In contrast to the report by Mireles ef al. [9], there was no effect of 25-(OH)D3 on the incidence of TD in birds raised on litter. The incidence of TD was low in this experiment, but severity was still decreased by B-(OH)D3. Genetics reportedly has a role in the effectiveness of 25-(OH)D3 and other vitamin D metabolites in preventing TD in broilers [12]. When added to a calcium-adequate diet containing 27.5 pg/kg cholecalciferol, 5 puglkg 25-(OH)D3 reduced TD and 10 pgkg maximized bone ash in chicks selected for a low incidence of TD. Zhang et al. [13] also reported a decrease in the incidence of TD in broiler chickens selected for a low incidence of TD when Z-(OH)D3 was fed, but no response was observed in chickens bred for a high incidence of TD. The objective of this study was to determine the effects of graded levels of 25-(OH)D3
55
56
rinized samples for measurement of phosphorus [24] and calcium [25] by colorimetric procedures. Excreta samples were collected from each pen in each experiment and saved for measurement of chromic oxide [26] and phytate phosphorus [27 in order to calculate phytate phosphorus retention values [B]. The experiments were terminated when the birds reached 17 days of age. All chicks were killed by cervical dislocation after body weight determination. The right tibia of each chick was scored for incidence and severity of TD by making a longitudinal cut across the proximal end of the bone [23]. The severity scores of TD ranged from 0 (no lesion) to 3 (severe lesion). The left tibia was saved for bone ash analysis [29]. The data were analyzed as described in previous publications [30,31, 32,331. In Experiment 1,both treatments containing 0 pgkg 25-(OH)D3 were analyzed as one treatment since there was adequate cholecalciferol in each treatment and there were no differences in any parameter measured.
RESULTS AND DISCUSSION There was no effect on body weight in either experiment (Tables 2 and 3). This agrees with the results of Edwards [l].However, Yarger et al. [lo] reported that 69 pgkg 25-(OH)D3 in place of a similar amount of cholecalciferol increased body weight of chickens grown to about 50 days of age in nine of 10 floor pen experiments. There was no difference in body weight in the one experiment in which 25-(OH)D3 was not effective. Rennie and Whitehead [11] did not observe a difference in body weight of 3-wk-old chicks grown in wire cages. However, they observed a decrease in 3- and 6-wk-old chickens fed 75pg/kg 25-(OH)D3 in place of cholecalciferol. Mireles et al. [9] found inconsistent results of 25-(OH)D3 on body weight of broilers grown to 7 wk of age on litter. Body weight was increased in one experiment, but decreased in another when 69 pgkg was fed. Feed efficiency was also not affected by 25-(OH)D3 in these studies. Yarger et al. [lo] reported better feed efficiency with 69 pgkg Z-(OH)D3. However, no feed efficiency responses were found by others [1, 11,121. There were no effects of 25-(OH)D3 on plasma calcium or phosphorus (Table 3). Rennie and Whitehead [ll] did not observe significant effects on plasma calcium and
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Crystalline 25-(OH)D3 was donated [19] and diluted to 140 mgkg with rice hulls. This premix was used to make the appropriate additions to the experimental diets. Another treatment consisted of supplemental cholecalciferol at 69p& to see if additional cholecalciferol yielded any response to 25-(OH)D3. Crystalline cholecalciferol [20] was diluted with rice hulls to 140 mgkg also. The experimental diets were fed to four pens of eight chicks per treatment. The horizontal mixer used to mix the diets was cleaned and sand was run through the mixer and auger that delivered the feed after mixing. Although cholecalciferol was not added to the premix, the complete basal diet was analyzed [21] to contain adequate ( > 20,ugkg) vitamin D activity [Z]. The mixer was also being used at this time to mix experimental swine diets containing a very high (25Opgkg added) level of cholecalciferol. Chromic oxide was added at 0.1% as an external indicator to determine phytate phosphorus retention values. The second experiment utilized 240 straight run chicks from the same source as Experiment 1. In Experiment 2, four treatments were allocated to six pens of 10 chicks each fed a low-calcium, TD-inducing diet as described by Edwards and Veltmann [23]. Crystalline cholecalciferol or 25-(OH)D3 was diluted in propylene glycol for liquid addition to the feed to see if the form of 25-(OH)D3 affected the results by supplying a more consistent source under the conditions of the study. The treatments were as follows: 1) 7 0 p g k g cholecalciferol, 2) 70 pgkg 25-(OH)D3, 3) 250 p g k g 25-(OH)D3, and 4) 250pg/kg 25-(OH)D3 added in dry form using the premix described previously. Analysis of the control diet prior to cholecalciferol supplementation indicated that vitamin D activity was present in the mixed feed at approximately 22pg (880 ICU)/kg. The brooder was placed in a room located at the dairy unit of the Martin Experiment Station, University of Tennessee, Martin, TN. Dark plastic was placed over all outside windows in the room. Clear plastic was placed over fluorescent lights (no diffusers) located on the ceiling. Lighting was continuous in the room and the battery-brooder. At 16 days of age, blood samples were obtained by cardiac puncture from one chick per pen. Plasma was extracted from the hepa-
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even when TD incidence is low. However, considering that higher levels of 25-(OH)D3 used in both experiments in this study did not influence the severity of TD in the chicks, it cannot be concluded from this study that 25-(OH)D3 can effectivelydecrease the severity of TD. There was no consistent effect of 25(OH)D3 on phytate phosphorus utilization in these experiments. In Experiment 2, there was a decrease (P < .OS). in phytate phosphorus retention when 25-(OH)D3 was fed, but no effect was observed in Experiment 1.Previous unpublished work in this laboratory found no effect of dietary E-(OH)D3 on phytate phosphorus utilization. Although 1,25(OH)2D3has been shown to increase phytate phosphorus retention [13, 34, 351, the author is not aware of any reports on the effect of E-(OH)D3 on this parameter. Rennie and Whitehead [ l l ] hypothesized that high levels of 25-(OH)D3 may affect chondrocyte differentiation by binding to the vitamin D receptor and activating Dresponsive genes or by inducing endogenous synthesis of l-hydroxylated metabolites. Yarger et al. [36] did not observe a change in serum 1,25-(OH)zD3 when 25-(OH)D3 was fed at 207 pgkg. However, there appears to be no relationship between blood levels of 1,25-(OH)@3 and the incidence of TD in broilers [5,12,34,37l. Studies on the relationship between genetics and nutrition [12, 131 suggest that when broiler chickens are bred for a low incidence of TD, additives such as 25-(OH)D3 may be effective because of their interaction with other causes of the cartilage abnormality- causes related to nutrition or to some factor other than genetics. Although researchers report that dietary 25-(OH)D3 may alleviates TD in broilers, there was no indication in these studies of any effect in chicks raised in cages.
CONCLUSIONS AND APPLICATIONS 1. Supplementation of 25-(OH)D3 at levels as high as 250pglkg did not prevent TD or affect growth performance in commercial broiler chicks raised in a battery-brooder. 2. Level of dietary calcium and form of 25-(OH)D3 addition did not affect the response observed with supplementation of 25-(OH)D3.
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phosphorus when high levels of 25-(OH)D3 alone were added to the diet. There was no indication that 25-(OH)D3 was able to reduce the incidence of TD in broiler chicks regardless of dietary calcium level or form of Z-(OH)D3 addition (Tables 2 and 3). This agrees with the data of Edwards [l]and one of two experiments reported by Rennie and Whitehead [ll]. In the experiment in which Rennie and Whitehead [ll]did not see a response, the incidence of TD in the chicks fed cholecalciferolwas 28%. The incidence of TD in the control diet was 34% in Experiment 1and 24% in Experiment 2 in this study. When 25-(OH)D3 was found to be effective by Rennie and Whitehead [ll], the incidence of TD in chicks fed cholecalciferol was 64%. Mitchell and Edwards [12] observed a decrease in TD incidence and severity when 25-(OH)D3 was added to a calcium-adequate (0.95 %) and cholecalciferol- a de quat e (27.5pgkg) diet fed to chicks bred for a low incidence of TD. The incidence of TD in this line when Z-(OH)D3 was not added was 44 and 61%, respectively, in the two experiments in which25-(OH)D3 was studied. Hence, from their studies, it appears that a high incidence of TD must be present to elicit a response from dietary25-(OH)D3.However, Mireleset al. [9] reported a significant decrease in TD incidence when lowered from 30 to 13%. Mireles etal. [9] utilized 500 birds per treatment, which is much higher than the number of chicks used in previous studies [ll,121or the present experiments. The severity of TD was generally not affected by 25-(OH)D3 (Tables 2 and 3). However, there was a significant decrease in the severity of TD when 46 pg/kg 25-(OH)D3 was fed compared to the diets in which no 25-(OH)D3 was fed in Experiment 1.Rennie and Whitehead [ll]found that 75 pgkg Z-(OH)D3 could decrease TD in broilers
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REFERENCES AND NOTES 1. Edwards, H.M., Jr., 1989. The effect of dietary cholecalciferol, 25-hydroxycholecalciferol and 1,25dihydroxycholecalciferol on the development of tibial dyschondroplasia in broiler chickens in the absence or presence of disulfiram. J. Nutr. 119647-652. 2. Edwards, H.M., Jr., 1990. Efficacy of several vitamin D compounds in the prevention of tibial dyschondroplasia in broiler chickens. J. Nutr. 12010541061.
4. Rennie, J.S., C.C. Whitehead, and B.H. Thorp, 1993. The effect of dietary 1,25dihydroxycholecalciferol in preventing tibial dyschondroplasia in broilers fed on diets imbalanced in calcium and phosphorus. Br. J. Nutr. 69:809-816.
5. Roberson, K.D. and EM.Edwards,Jr., 1996. Effect of dietary 1,2.5-dihydroxycholecalciferollevel on broiler performance. Poultry Sci. 7590-94. 6. Rennie, J.S., H A McCormaclc, C. Farquharson, J.L Berry, EB. Mmer, and C.C. Whitehead, 1995.Interaction between dieta 1,25dihydroxycholecalciferol and calcium and effects ormanagement on the occurrence of tibial dyschondroplasia, le abnormalities, and performance in broiler chickens. br. Poultry Sci. 36:4654?7. 7. Fuqnhprson, C., kS.Law, E Seawright, D.W. Burt, and C.C. Whitehead, 1993. In y i y ~effect of 1,25dihydroxycholecalciferol on the proliferation and differentiation of avian chondrocytes. J. Bone and Min. Res. 81081-1088. 8. Hughes, M.R, D.J. Baylink, P.G. Jones, and M.R Haossler, 1976. Radioligand receptor assay for 25hydraxyvitamin Dm3. Application to hypervitaminosis D J . Clin. Invest. 5861-70.
9. Mireles, A, Jr., S. Kim, B. Krautmann, J. Yarger, and L Siark, 1996. Effect of 25-hydroxycholecalciferol (z-(OH)D3) on broiler field performance and incidence of tibial dyschondro lasia(TD): Minimum D3 metabolite consumption periodq Poultry Sci. 75(Suppl):280 (Abs). 10. Yarger, J.G., CA. Saunders, J.L McNaughton, C.L Quarles, B.W.Hollis, and RW. Gray, 1995. Comparison of dietary25-hydro holecalciferol and cholecalciferol in broiler chickens. %tty Sci. 741159-1167. 11.Rennie, J.S. and C.C. Whitehead,1996. Effectiveness of dietary 25- and 1-hydroxycholecalciferolin combating tibial dyschondroplasia in broiler chickens. Br. Poultry Sci. 37413-421. 12. Mitchell, RD. and H.M. Edwards,Jr., 1997. The effects of ultraviolet light and cholecalciferol and its metabolites on the development of leg abnormalities in chickens genetically selected for a high or low incidence of tibial dyschondroplasia. Poultry Sci. 76346354. 13. Zhang, X,G. Uu,G.R McDaniel, and D.A. Roland, 1997. Responses of broiler lines selected for tibial dyschondroplasia incidence t o s u p lementary 25hydroxycholecalciferol.J. Appl. Poultrykes. 6410-416. 14. McNaoghton, J.L, EJ. Day, and B.C. Dilworlh, 1977. T h e chick's requirement for 25-hydroxy-
15. McNaughton, J.L and R Murray, 1990. Effect of 2.5-hydroxycholecalciferol (25-(OH)D!) and vitamin D3 on hos horus requirement of broilers. Poultry Sci. 69({ uppl .178 (Abs).
7.
16. Edwards, H.M., Jr., 1993. Dietary 1,25-dihydroxycholecalciferol supplementation increases h ate phosphorus utilization in chickens. J. Nutr. 123!6&7. 17. Seaboard Farms, Mayfield, KY 42066. 18. Petersime Incubator Co., Gettysburg, OH 45328.
19. Amoco BioProducts Corporation, Napenille, IL 60563. 20. Amersham, Arlington Heights, 1L 6ooo5.
21. Woodson-Tenent Laboratories, Inc., Memphis, TN 38101. 2 2 Edwards,H.M., Jr., M.A. Elliot, S. Sooncharernyin& and W.M. Brillon, 1994. Quantitative requirement
for cholecalciferol in the absence of ultraviolet light. Poultry Sci. 73288-294. 23. Edwards, H.M., Jr. and J . R Vellmann, 1983. The role of calcium and phosphorus in the etiology of tibial dyschondroplasia in young chicks. J. Nutr. 1131568-1575. 24. Goldenberg, H. and A. Fernandeq 1966. Simplified method for the estimation of inorganic phosphorus in body fluids. Clin. Chem. 128714382.
25. Moorehead, W.R and H.G. B b 1974.2-amino2-methyl-1-propanol as the alkalizing agent in an improved continuous flow cresolphtalein com lexone procedure for calcium in serum. Clin. Chem. &:14581460. 26. Fenlon, T.W. and M. Fenton, 1979. An improved procedure for the determination of chromic oxide in feed and feces. Canadian J. Anim. Sci. 59:631-634. 27. Common, RH., 1940. The phytic acid content of some poultry feeding stuffs. Analyst 65:79-83. 28. Edwards,H.M., Jr. and M.B. Gillis, 1959. A chromic oxide balance method for determining phosphate availability. PoultIy Sci. 38569-574. 29. Associalion of Ol'f'icial Analytical Chemists, 1995. Official Methods of Analysis. 16th Edition. Assn. Offic. Anal. Chemists, Washington, DC. 30. The data were anal ed by ANOVA using the General Linear Models (GrM) rocedure of SAS [31]. Regression analysis of the 25-(8H)D3 level effect was evaluated. Treatment means were separated by Duncan's multiple range test [32] with a 5% level of probability, Orthogonal contrasts were conducted when more powerful statistical comparisons of treatment effects were desired [33].
31. SAS Institute, 1985. SAS User's Guide: Statistics. Version 5 Edition. SAS Institute, Inc., Cary, NC.
32. Duncan, D.B., 1955. Multiple range and multiple
F tests. Biometrics 11:142. 33. Sleel, RG.D. and J.H. Torrie, 1980. Princi les and Procedures of Statistics. 2nd Edition. McGraw-dl1 Book Co.,New York,NY.
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3. Edwards, H.M., Jr., M.A. Ellfol, a n d S. Sooncharernyhg, 1992. Effect of dietary calcium on tibial dyschondro lasia. Interaction with light, cholecalciferol, 1,25-di~ydroxycholecalciferol, protein, and synthetic zeolite. Poultry Sci. 71:2041-2055.
cholecalciferol and cholecalciferol. Poultry Sci. 56511516.
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61 36. Yarger, J.G., C.L Quarles, B.W. Hollis, andRW. Gray, 1995. Safety of 25-hydroxycholecalciferol as a source Of cholecalciferol in pOukry rations. Poultry SCi. 74~1437-1446.
34. Robcrson, K.D. and H.M. Edwards, Jr., 1994. Effects of ascorbic acid and 1,Zdihydroxycholecalciferol on alkaline phosphatase and tibial dyschondroplasia in broiler chickens. Br. Poultry Sci. 35:763-773.
ditive effectsof 1,25dihydro~cholecalciferoland phytase on phytate phosphorus utilization and related parameters in broiler carcasses. Poultry Sci. 75:111-119.
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and 1,Z-dihydroxycholecalciferolon the develGment of tibial dyschondroplasia in broilers during the starter and grower periods. Poultry Sci. 74:1495-1505.
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