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Evaluation of relative bioavailability of 25-hydroxycholecalciferol to cholecalciferol for broiler chickens.

INTRODUCTION

Cholecalciferol (vitamin [D.sub.3]) has been used as a feed additive to regulate calcium (Ca) and phosphorus (P) metabolism and bone development in animals for many years. Vitamin [D.sub.3] undergoes 25-hydroxylation in animal livers to transform 25-hydroxycholecalciferol (25-OH-[D.sub.3]). The commercial 25-OH-[D.sub.3] has been produced and approved for use in poultry and swine feed in China in 2014.

Previous research has shown that body weight and feed efficiency of broiler chickens fed with 25-OH-[D.sub.3] were greater than those of the birds fed with vitamin [D.sub.3] (Yarger et al., 1995; Fritts and Waldroup, 2003). Replacing vitamin [D.sub.3] by 25-OH-[D.sub.3] at 50% has a beneficial effect on the growth performance of broilers (Koreleski and Swiatkiewicz, 2005). These data indicate that the relative bioavailability (RBV) of 25-OH-[D.sub.3] is higher than that of vitamin [D.sub.3].

However, no consistent results have been obtained in the RBV of 25-OH-[D.sub.3] to vitamin [D.sub.3]. Soares et al. (1995) reviewed that the RBV of 25-OH-[D.sub.3] to vitamin [D.sub.3] ranged from 1.0 to 4.0 when Ca absorption, plasma Ca, bone ash, bone strength, and tibial dyschondroplasia were used as the criteria. Atencio et al. (2005) found that the RBV values of 25-OH-[D.sub.3] to vitamin [D.sub.3] were 1.38, 1.33, 1.28, and 1.11 for egg production, hatchability, late embryo mortality, and body ash of the progeny in broiler breeder hen diets, respectively. These data reveal the differences in the RBV of 25-OH-[D.sub.3] to vitamin [D.sub.3] among the studies. The RBV of 25-OH-[D.sub.3] to vitamin [D.sub.3] in poultry diets should be further clarified.

Researchers usually use the tibia to evaluate the RBV of vitamin D derivatives. In fact, the differences in growth and mineralization among the femur, tibia, and metatarsus in poultry have been observed (Applegate and Lilburn, 2002; Goetting-Fuchs et al., 2012; Han et al., 2015). The femur and metatarsus should also be used as criteria to evaluate the RBV of 25-OH-[D.sub.3] to vitamin [D.sub.3].

Therefore, the present study was conducted to investigate the effects of 25-OH-[D.sub.3] and vitamin [D.sub.3] on the growth performance and development of the femur, tibia, and metatarsus and to re-evaluate the RBV of 25-OH-[D.sub.3] to vitamin [D.sub.3] in broilers fed with Ca- and P-deficient diets.

MATERIALS AND METHODS

Birds, diets, and management

All of the procedures adopted in this study were approved by the Animal Care Committee of Shangqiu Normal University.

On the day of hatch, 450 female Ross 308 broiler chickens were assigned to nine treatments, with five replicates of ten birds each. The initial body weight of broiler chickens was 46.5[+ or -]1.9 g. Birds from 1 to 13 d of age were reared in stainless steel starter cages (70 cmx70 cmx30 cm). At 14 d, the broilers were transferred to stainless steel grower cages (190 cmx50 cmx35 cm). The basal diet contained 0.50% Ca and 0.25% non-phytate phosphorus (NPP) and was not supplemented with vitamin D. Vitamin [D.sub.3] was fed at 0, 2.5, 5.0, 10.0, and 20.0 [micro]g/kg and 25-OH-[D.sub.3] was fed at 1.25, 2.5, 5.0, and 10.0 [micro]g/kg. The birds were provided mash diet (Table 1) and water ad libitum. The lighting system consisted of 23 h of light from 1 d to 3 d and 20 h of light from 4 d to 21 d. Room temperature was controlled at 33[degrees]C from 0 d to 3 d, 30[degrees]C from 4 d to 7 d, 27[degrees]C from 8 d to 14 d, and 24[degrees]C from 15 d to 21 d.

25-OH-[D.sub.3] and vitamin [D.sub.3]

Crystalline 25-OH-[D.sub.3] (98%) and vitamin [D.sub.3] (99%) were supplied by Changzhou Book Chemical Co., Ltd. (Changzhou, China) and Jiaxing Tianhecheng Biological Technology Co., Ltd. (Jiaxing, China), respectively. The 25OH-[D.sub.3] and vitamin [D.sub.3] solutions were prepared by the method of Biehl and Baker (1997). Crystalline 25-OH-[D.sub.3] and vitamin [D.sub.3] were weighed and dissolved in ethanol. Then, they were diluted to a final concentration of 10 mg/L of 25-OH-[D.sub.3] or vitamin [D.sub.3] in a solution of 5% ethanol and 95% propylene glycol. After the preparation, the solution of 25-OH-[D.sub.3] or vitamin [D.sub.3] was added to the diets.

Sample collection

The birds were weighed on d 21 after 12 h of fasting. Ten chicks per treatment were randomly selected for the collection of blood, femur, tibia, and metatarsus. Plasma samples (5 mL) were collected through cardiac puncture and centrifuged for 10 min at 3,000xg at 20[degrees]C. The birds were killed after collecting the blood samples. The femur, tibia, and metatarsus of the individual birds were excised and frozen at -20[degrees]C for analysis.

Sample analysis

Plasma Ca and inorganic phosphorus (Pi) were determined using a Shimadzu CL-8000 analyzer (Shimadzu Corp., Kyoto, Japan) following the instructions of the manufacturer.

The left femur, tibia, and metatarsus were boiled for 5 min to loosen the muscle tissues using the method of Hall et al. (2003). The meat, connective tissue, and fibula bone were completely removed using scissors and forceps. The bones were placed in a container of ethanol for 24 h (removing water and polar lipids) after cleaning. Afterward, the bones were further extracted in anhydrous ether for 24 h (removing non-polar lipids). The bones were dried at 105[degrees]C for 24 h before weighing. The bone ash content was determined by ashing the bone in a muffle furnace for 48 h at 600[degrees]C.

The right tibia was utilized to analyze the breaking-strength, which was determined using an all-digital electronic universal testing machine (Shenzhen Hengen Instrument Co. Ltd., Shenzhen, China). The tibias were cradled on two support points measuring 4 cm apart. Force was applied to the midpoint of the same face of each tibia using a 50 kg load cell with a crosshead speed of 10 mm/min (Jendral et al., 2008).

Dietary and bone Ca were determined by the ethylene diamine tetraacetic acid titration method, and P was determined by photometric methods after reaction with ammonium molybdate and ammonium metavanadate (Han et al., 2013).

Statistical analysis

Replicate means are the experimental units in the statistical analysis. Data were analyzed with a general linear model of the SAS software (SAS Institute, 2002). The RBV of 25-OH-[D.sub.3] was determined using vitamin [D.sub.3] as the standard source by the slope ratio method (Littell et al., 1997). Feed intake differed for the dietary treatments so that vitamin D intake rather than vitamin D content was used as the independent variable for regression analysis. The model is as follows: y = a+[b.sub.1][x.sub.1]+ [b.sub.2][x.sub.2], where y is the response, [x.sub.1] is vitamin [D.sub.3] intake, [x.sub.2] is 25-OH-[D.sub.3] intake, a is the intercept, and [b.sub.1] and [b.sub.2] are the slope of vitamin [D.sub.3] and 25-OH-[D.sub.3], respectively. Orthogonal comparisons were performed to determine the linear and quadratic effects of the 25-OH-[D.sub.3] or vitamin [D.sub.3] levels on growth performance and bone mineralization. The basal diet treatment was included for both vitamin [D.sub.3] and 25-OH-[D.sub.3] when conducting orthogonal polynomial contrast test. Means were compared by conducting Tukey test when probability values were significant (p<0.05).

RESULTS

Growth performance

Dietary 25-OH-[D.sub.3] linearly affected body weight gain (BWG), feed intake (FI), feed efficiency, and mortality in 1to 21-d-old broiler chickens (p<0.05, Table 2). Vitamin [D.sub.3] levels also influenced the above parameters (p<0.05). 25OH-[D.sub.3] or vitamin [D.sub.3] did not affect the plasma Ca or Pi concentration (p>0.05).

Bone mineralization

The femur, tibia, and metatarsus are three leg bones in poultry. They reflect the body bone quality of birds. The linear relationships between the level of 25-OH-[D.sub.3] or vitamin [D.sub.3] and the weight, length, ash weight, and the percentage of ash, Ca, and P of the femur in broiler chickens were observed (p<0.05, Table 3).

Dietary 25-OH-[D.sub.3] or vitamin [D.sub.3] linearly improved the tibia breaking-strength (p<0.05, Table 4). Similar results were observed in the relationship between the level of 25OH-[D.sub.3] or vitamin [D.sub.3] and the weight, length, ash weight, and the percentage of ash, Ca, and P of the tibia.

Increasing the 25-OH-[D.sub.3] or vitamin [D.sub.3] level linearly increased the weight, length, ash weight, and the percentage of ash, Ca, and P of the metatarsus (p<0.05, Table 5).

Relative bioavailability of 25-OH-[D.sub.3] to vitamin [D.sub.3]

The slope ratio method was used to evaluate the RBV of 25-OH-[D.sub.3] to vitamin [D.sub.3] in broiler chickens (Table 6). Vitamin [D.sub.3] and 25-OH-[D.sub.3] intake was used as the independent variable for regression analysis. The slopes of vitamin [D.sub.3] and 25-OH-[D.sub.3] were 16.706 and 30.862, respectively, when BWG was used as the criterion. Thus, the RBV value of 25-OH-[D.sub.3] to vitamin [D.sub.3] was 1.85 (namely 185%, 30.862 divided by 16.706).

Using BWG as the criterion, the RBV value of 25-OH[D.sub.3] to vitamin [D.sub.3] was 1.85. When the weight, length, ash weight, and the percentage of ash, Ca, and P of the femur were used as the criteria, the RBV of 25-OH-[D.sub.3] to vitamin [D.sub.3] were 1.88, 1.82, 2.00, 2.03, 2.45, and 2.22, respectively. Using the same parameters of the tibia as the criteria, the RBV of 25-OH-[D.sub.3] to vitamin [D.sub.3] were 2.12, 1.86, 2.17, 2.13, 2.52, and 2.52, respectively. Metatarsus mineralization was also used as a criterion. The above RBV values were 2.05, 1.89, 2.00, 1.76, 1.73, and 1.65, respectively.

Generally, the bioavailability of 25-OH-[D.sub.3] is higher than that of vitamin [D.sub.3] in broilers. The average RBV of 25OH-[D.sub.3] to vitamin [D.sub.3] is approximately 2.03 (namely 203%) in promoting growth performance and bone mineralization in 1- to 21-d-old broiler chicken diets.

DISCUSSION

Previous research has shown that vitamin [D.sub.3] levels linearly improve growth and bone quality when broilers are fed with Ca- and NPP-deficient diets; by contrast, growth is quadratically or not significantly affected by vitamin [D.sub.3] levels when the Ca and NPP contents are sufficient (Aburto et al., 1998; Baker et al., 1998; Rao et al., 2009). Thus, the Ca- and NPP-deficient diet was designed in the present study. The optimal dietary Ca to NPP ratio is 2.0 to promote growth performance and bone mineralization in broiler chickens (Bar et al., 2003; Rao et al., 2007). Therefore, the Ca and NPP levels were 0.50% and 0.25%, respectively.

In the comparison of 25-OH-[D.sub.3] to vitamin [D.sub.3] at the 3.125 [micro]g/kg level, the RBV of 25-OH-[D.sub.3] to vitamin [D.sub.3] ranged from 1.08 to 4.00 in broiler breeder hens (Atencio et al., 2005). However, no statistical differences in the performance between 25-OH-[D.sub.3] and vitamin [D.sub.3] were observed when their level reached 12.5 [micro]g/kg in hens (Atencio et al., 2005) or 50 gg/kg in broiler chickens (Fritts and Waldroup, 2003). These data indicate that the 25-OH[D.sub.3] or vitamin [D.sub.3] level should not exceed their requirement when evaluating the RBV of 25-OH-[D.sub.3] to vitamin [D.sub.3].

The optimal level of 25-OH-[D.sub.3] was 10 [micro]g/kg for promoting bone ash in broiler chicken diets (Goodgame et al., 2011). Thus, the level of 25-OH-[D.sub.3] ranged from 1.25 to 10 [micro]g/kg in the present study. No significant differences were observed in the tibia ash, breaking-strength, and the contents of Ca and P among the broilers fed with vitamin [D.sub.3] ranging from 25 to 1,000 [micro]g/kg (Han et al., 2013). Therefore, vitamin [D.sub.3] levels were lower than those of our previous research (Han et al., 2013) and were at 2.5, 5, 10, and 20 [micro]g/kg in the present study. Our results revealed the linear relationship between the 25-OH-[D.sub.3] or vitamin [D.sub.3] level and the performance or bone mineralization in broiler chickens.

The positive effects of 25-OH-[D.sub.3] or vitamin [D.sub.3] on growth performance and tibia weight, breaking-strength, and the percentage of ash, Ca, and P in broiler chickens have been observed (Fritts and Waldroup, 2003; Rao et al., 2006; Han et al., 2012). The improvement of vitamin D derivatives on performance and bone of birds was caused by the increase of Ca and P utilization. Research has shown that addition of vitamin [D.sub.3] (Qian et al., 1997), 25-OH-[D.sub.3] (Ledwaba and Roberson, 2003), 1[alpha]-OH-[D.sub.3] (Shirley, 2003; Han et al., 2012), or 1,25-[(OH).sub.2]-[D.sub.3] (Edwards, 2002) improves the retention of Ca and P in broiler chicken diets.

Previous research has shown that when bone ash was used as criteria, the RBV of 25-OH-[D.sub.3] to vitamin [D.sub.3] ranged from 1.0 to 2.5 (Soares et al., 1995). The RBV was from 1.11 to 1.38 in broiler breeder hens (Atencio et al., 2005). The present study showed that the RBV of 25-OH-[D.sub.3] to vitamin [D.sub.3] is approximately 2.03 for promoting growth performance and bone mineralization in 1- to 21-d-old broiler chickens. The differences in RBV values among the criteria were observed. Tibia gave the highest RBV of 25OH-[D.sub.3] to vitamin [D.sub.3] (2.22) and followed by femur (2.07). The BWG and metatarsus criteria yielded the lowest values (1.85).

CONCLUSIONS

The present study indicates that the average RBV of 25OH-[D.sub.3] to vitamin [D.sub.3] is approximately 2.03 (namely 203%) for promoting growth performance and bone mineralization in 1- to 21-d-old broiler chickens fed with Ca- and P-deficient diets.

http://dx.doi.org/10.5713/ajas.15.0553

CONFLICT OF INTEREST

We certify that there is no conflict of interest with any financial organization regarding the material discussed in the manuscript.

ACKNOWLEDGMENTS

This study was supported by the National Natural Science Foundation of China (31101732), the Innovation Scientists and Technicians Troop Construction Projects of Henan Province, the Foundation of the Education Department of Henan Province (16A230003), and the Shangqiu Normal University Foundation (2013GGJS10).

REFERENCES

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J. C. Han1 (2),*, G. H. Chen (1,2), J. G. Wang (1,2), J. L. Zhang (1,2), H. X. Qu (1), C. M. Zhang (1), Y. F. Yan (1),*, and Y. H. Cheng (3)

(1) Department of Animal Science, College of Life Science, Shangqiu Normal University, Shangqiu, Henan 476000, China

* Corresponding Authors: J. C. Han. Tel: +86-0370-2592849, E-mail: j.c.han@hotmail.com / Y F. Yan. Tel: +86-0370-2594568, E-mail: yanyf01@sina.com

(2) College of Animal Husbandry and Veterinary Science, Henan Agricultural University, Zhengzhou, Henan 450002, China.

(3) Department of Biotechnology and Animal Science, National Ilan University, I-Lan 26047, Taiwan.

Submitted Jul. 2, 2015; Revised Aug. 25, 2015; Accepted Oct. 14, 2015
Table 1. Ingredients and nutrient composition of the basal diet

Items                                   Basal diet

Ingredient (%)
  Corn                                    60.73
  Soya bean meal (43% CP)                 32.00
  Soya bean oil                            1.60
  Soya bean protein isolate (65% CP)       3.47
  Limestone                                0.67
  Dicalcium phosphate                      0.71
  L-lysine HCl (98%)                       0.14
  DL-methionine (98%)                      0.14
  Trace mineral premix (1)                 0.01
  Vitamin premix (2)                       0.03
  Choline chloride (50%)                   0.20
  Sodium chloride                          0.30
Nutrient composition
  Metabolizable energy (kcal/kg)         2,975.20
  Crude protein (CP, %)                   21.24
  Analyzed calcium (Ca, %)                 0.52
  Analyzed total phosphorus (TP, %)        0.49
  Non-phytate phosphorus (NPP, %)          0.25

(1) The trace mineral premix provided the following
(per kg of diet): 80 mg iron; 40 mg zinc; 8 mg copper;
60 mg manganese; 0.35 mg iodine; and 0.15 mg selenium.

(2) The vitamin premix provided the following
(per kg of diet): 8,000 IU vitamin A; 20 IU vitamin E;
0.5 mg menadione; 2.0 mg thiamine; 8.0 mg riboflavin;
35 mg niacin; 3.5 mg pyridoxine; 0.01 mg vitamin B12;
10.0 mg pantothenic acid; 0.55 mg folic acid;
and 0.18 mg biotin.

Table 2. Effects of vitamin D3 and 25-OH-D3 on growth performance
and plasma mineral concentration in 1- to 21-d-old broiler
chickens

                                                 Growth

Vitamin [D.sub.3]     25-OH-[D.sub.3]   BWG (g/bird)   FI (g/bird)
([micro]g/kg)          ([micro]g/kg)

0                            0            233 (d)        431 (d)
2.5                                       338 (c)       621 (bc)
5.0                                       433 (b)        709 (b)
1 0.0                                     584 (a)        932 (a)
20.0                                      602 (a)        955 (a)
                           1.25           321 (c)        585 (c)
                            2.5           426 (b)        702 (b)
                            5.0           556 (a)        866 (a)
                           1 0.0          574 (a)        927 (a)
SEM                                          19            27
p value
  Vitamin [D.sub.3]       Linear           <0.001        <0.001
                         Quadratic         0.002          0.006
  25-OH-[D.sub.3]         Linear           <0.001        <0.001
                         Quadratic         0.036          0.089

                                                 Growth

Vitamin [D.sub.3]     25-OH-[D.sub.3]   FE (BWG/FI)   Mortality (%)
([micro]g/kg)          ([micro]g/kg)

0                            0           0.542 (c)       26 (a)
2.5                                     0.548 (bc)       12 (b)
5.0                                      0.612 (a)        0 (b)
1 0.0                                    0.627 (a)        0 (b)
20.0                                     0.631 (a)        0 (b)
                           1.25         0.549 (bc)       10 (b)
                            2.5         0.606 (ab)        0 (b)
                            5.0          0.642 (a)        0 (b)
                           1 0.0         0.619 (a)        0 (b)
SEM                                        0.007            2
p value
  Vitamin [D.sub.3]       Linear          <0.001         <0.001
                         Quadratic         0.363          0.010
  25-OH-[D.sub.3]         Linear          <0.001         <0.001
                         Quadratic         0.067         <0.001

                                                Plasma

Vitamin [D.sub.3]     25-OH-[D.sub.3]   Ca (mg/dL)   Pi (mg/dL)
([micro]g/kg)          ([micro]g/kg)

0                            0             5.45         3.07
2.5                                        6.69         3.27
5.0                                        6.90         3.83
1 0.0                                      6.93         4.82
20.0                                       6.98         4.64
                           1.25            5.72         3.24
                            2.5            5.70         3.60
                            5.0            6.70         3.66
                           1 0.0           6.73         4.17
SEM                                        0.16         0.18
p value
  Vitamin [D.sub.3]       Linear          0.043        0.022
                         Quadratic        0.171        0.890
  25-OH-[D.sub.3]         Linear          0.013        0.038
                         Quadratic        0.732        0.797

25-OH-[D.sub.3], 25-hydroxycholecalciferol; BWG,
body weight gain; FI, feed intake; FE, feed efficiency;
Ca, calcium; Pi, inorganic phosphorus;
SEM, standard error of the mean.

(a-d) Means in the same column without a common
superscript differ significantly (p<0.05).

Table 3. Effects of vitamin [D.sub.3] and 25-OH-[D.sub.3]
on femur mineralization in 1- to 21-d-old broiler chickens

Vitamin [D.sub.3]     25-OH-[D.sub.3]   Weight (g)    Length (cm)
([micro]g/kg)             (gg/kg)

0                            0           0.56 (d)      3.43 (f)

2.5                                      0.63 (cd)     3.75 (ef)
5.0                                      0.78 (b)     4.34 (bcd)
1 0.0                                    1.01 (a)      4.69 (ab)
20.0                                     1.07 (a)      4.87 (a)
                           1.25          0.69 (bc)    3.85 (def)
                            2.5          0.80 (b)     4.10 (cde)
                            5.0          0.98 (a)     4.58 (abc)
                           1 0.0         1.02 (a)      4 77 (ab)
SEM                                        0.03          0.08
p value
  Vitamin [D.sub.3]       Linear          <0.001        <0.001
                         Quadratic         0.352         0.268
  25-OH-[D.sub.3]         Linear          <0.001        <0.001
                         Quadratic         0.424         0.590

Vitamin [D.sub.3]     25-OH-[D.sub.3]     Ash (g)       Ash (%)
([micro]g/kg)             (gg/kg)

0                            0           0.17 (d)      26.64 (e)
2.5                                      0.20 (d)          0
5.0                                      0.29 (c)     37.55 (bc)
1 0.0                                    0.40 (ab)     39.55 (b)
20.0                                     0.45 (a)      43.71 (a)
                           1.25          0.23 (cd)     33.07 (d)
                            2.5          0.29 (c)     36.37 (bcd)
                            5.0          0.38 (b)      38.64 (b)
                           1 0.0         0.45 (a)      44.14 (a)
SEM                                        0.02          0.81
p value
  Vitamin [D.sub.3]       Linear          <0.001        <0.001
                         Quadratic         0.288         0.033
  25-OH-[D.sub.3]         Linear          <0.001        <0.001
                         Quadratic         0.457         0.368

Vitamin [D.sub.3]     25-OH-[D.sub.3]     Ca (%)         P (%)
([micro]g/kg)             (gg/kg)

0                            0           10.30 (e)     4.92 (f)
2.5                                     12.70 (cd)     6.04 (de)
5.0                                     13.01 (cd)    6.78 (cde)
1 0.0                                   14.02 (abc)    7.13 (bc)
20.0                                    15.20 (ab)     7.88 (ab)
                           1.25         11. 31 (de)    5.91 (e)
                            2.5         13.48 (bc)    6.68 (cde)
                            5.0         14.52 (abc)    6.95 (cd)
                           1 0.0         15.86 (a)     8.22 (a)
SEM                                        0.28          0.16
p value
  Vitamin [D.sub.3]       Linear          <0.001        <0.001
                         Quadratic         0.318         0.207
  25-OH-[D.sub.3]         Linear          <0.001        <0.001
                         Quadratic         0.761         0.936

25-OH-[D.sub.3], 25-hydroxycholecalciferol;
SEM, standard error of the mean.

(a-f) Means in the same column without a common
superscript differ significantly (p<0.05).

Table 4. Effects of vitamin [D.sub.3] and 25-OH-[D.sub.3]
on tibia mineralization in 1- to 21-d-old broiler chickens

Vitamin [D.sub.3]      25-OH-       BS (N)     Weight (g)
([micro]g/kg)         [D.sub.3]
                       (gg/kg)

0                         0       20.28 (d)     0.66 (d)
2.5                               24.94 (d)     0.80 (d)
5.0                               39.87 (c)    1.01 (bc)
10.0                              54.10 (b)     1.32 (a)
20.0                              68.91 (a)     1.38 (a)
                        1.25      21.51 (d)    0.83 (cd)
                         2.5      39.83 (c)     1.06 (b)
                         5.0      49.32 (bc)    1.32 (a)
                        10.0      71.18 (a)     1.39 (a)
SEM                                  2.88         0.04
p value
  Vitamin [D.sub.3]    Linear       <0.001       <0.001
                      Quadratic     0.088        0.633
  25-OH-[D.sub.3]      Linear       <0.001       <0.001
                      Quadratic     0.009        0.282

Vitamin [D.sub.3]      25-OH-      Length      Ash (g)      Ash (%)
([micro]g/kg)         [D.sub.3]     (cm)
                       (gg/kg)

0                         0       4.71 (d)    0.18 (e)     26.82 (f)
2.5                               5.26 (c)    0.26 (d)    32.70 (de)
5.0                               5.71 (bc)   0.37 (c)    36.70 (cd)
10.0                              6.23 (a)    0.51 (b)    39.61 (abc)
20.0                              6.38 (a)    0.57 (ab)   41.65 (ab)
                        1.25      5.38 (c)    0.26 (d)     31.81 (e)
                         2.5      5.52 (c)    0.39 (c)    36.88 (cd)
                         5.0      6.12 (ab)   0.50 (b)    37.76 (bc)
                        10.0      6.30 (a)    0.59 (a)     42.93 (a)
SEM                                 0.09        0.02         0.78
p value
 Vitamin [D.sub.3]     Linear      <0.001      <0.001       <0.001
                      Quadratic     0.125       0.765        0.059
 25-OH-[D.sub.3]       Linear      <0.001      <0.001       <0.001
                      Quadratic     0.195       0.581        0.269

Vitamin [D.sub.3]      25-OH-       Ca (%)        P (%)
([micro]g/kg)         [D.sub.3]
                       (gg/kg)

0                         0        9.76 (e)      5.08 (d)
2.5                                11.51 (d)    6.00 (cd)
5.0                               13.18 (bc)    6.83 (bc)
10.0                              13.81  (bc)   7.14 (ab)
20.0                              14.13 (ab)    7.15 (ab)
                        1.25      11.23 (de)     5.53 (d)
                         2.5      12.44 (cd)    6.77 (bc)
                         5.0      13.14 (bc)    6.91 (abc)
                        10.0       15.59 (a)     7.77 (a)
SEM                                  0.27          0.14
p value
  Vitamin [D.sub.3]    Linear       <0.001        <0.001
                      Quadratic      0.003        0.015
  25-OH-[D.sub.3]      Linear       <0.001        <0.001
                      Quadratic      0.288        0.715

25-OH-[D.sub.3], 25-hydroxycholecalciferol;
BS, breaking-strength; SEM, standard error of the mean.

(a-f) Means in the same column without a common
superscript differ significantly (p<0.05).

Table 5. Effects of vitamin [D.sub.3] and 25-OH-[D.sub.3]
on metatarsus mineralization in 1- to 21-d-old broiler chickens

Vitamin [D.sub.3]      25-OH-     Weight (g)    Length (cm)
(gg/kg)               [D.sub.3]
                       (gg/kg)

0                         0        0.50 (c)      3.56 (f)
2.5                                0.58 (c)      3.84 (ef)
5.0                                0.80 (b)      4.29 (bc)
1 0.0                              0.91 (ab)     4.61 (ab)
20.0                               0.97 (a)      4.65 (a)
                        1.25       0.59 (c)      3.92 (de)
                         2.5       0.80 (b)      4.23 (cd)
                         5.0       0.94 (ab)    4.55 (abc)
                        1 0.0      0.94 (ab)    4.58 (abc)
SEM                                  0.03          0.06
p value
  Vitamin [D.sub.3]    Linear       <0.001        <0.001
                      Quadratic      0.133         0.052
  25-OH-[D.sub.3]      Linear       <0.001        <0.001
                      Quadratic      0.071         0.024

Vitamin [D.sub.3]      25-OH-       Ash (g)       Ash (%)
(gg/kg)               [D.sub.3]
                       (gg/kg)

0                         0        0.12 (c)      22.20 (e)
2.5                                0.17 (c)     29.02 (cd)
5.0                                0.25 (b)     31.69 (bcd)
1 0.0                              0.30 (ab)    33.06 (abc)
20.0                               0.35 (a)      36.71 (a)
                        1.25       0.16 (c)      27.41 (d)
                         2.5       0.25 (b)     29.50 (cd)
                         5.0       0.30 (ab)    31.65 (bcd)
                        1 0.0      0.34 (a)     35.86 (ab)
SEM                                  0.01          0.69
p value
  Vitamin [D.sub.3]    Linear       <0.001        <0.001
                      Quadratic      0.582         0.029
  25-OH-[D.sub.3]      Linear       <0.001        <0.001
                      Quadratic      0.519         0.593

Vitamin [D.sub.3]      25-OH-       Ca (%)         P (%)
(gg/kg)               [D.sub.3]
                       (gg/kg)

0                         0        7.68 (e)      3.67 (f)
2.5                                10.23 (d)     4.94 (de)
5.0                               11.77 (bc)    5.81 (abcd)
1 0.0                             12.01 (abc)   5.93 (abc)
20.0                               13.20 (a)     6.68 (a)
                        1.25       10.39 (d)     4.92 (e)
                         2.5      10.84 (cd)    5.40 (cde)
                         5.0      11.32 (cd)    5.67 (bcde)
                        1 0.0     12.88 (ab)     6.35 (ab)
SEM                                  0.25          0.14
p value
  Vitamin [D.sub.3]    Linear       <0.001        <0.001
                      Quadratic     <0.001         0.007
  25-OH-[D.sub.3]      Linear       <0.001        <0.001
                      Quadratic      0.076         0.104

25-OH-[D.sub.3], 25-hydroxycholecalciferol;
SEM, standard error of the mean.

(a-f) Means in the same column without a common
superscript differ significantly (p<0.05).

Table 6. Relative bioavailability (RBV) of 25-OH-[D.sub.3]
to vitamin [D.sub.3] based on vitamin D intake (gg/bird)
in 1- to 21-d-old broiler chickens with the slope
ratio method

                                           Slope [+ or -] SE

Criteria                     Intercept     Vitamin [D.sub.3]

Growth performance
  Weight gain                 334.545    16.706 [+ or -] 1.799
Femur mineralization
  Weight                       0.666      0.024 [+ or -] 0.003
  Length                       3.813      0.065 [+ or -] 0.008
  Ash weight                   0.216      0.014 [+ or -] 0.001
  Ash percentage              32.147      0.669 [+ or -] 0.075
  Ca percentage               11.776      0.198 [+ or -] 0.030
  P percentage                 5.797      0.121 [+ or -] 0.015
Tibia mineralization
  Weight                       0.838      0.033 [+ or -] 0.004
  Length                       5.242      0.070 [+ or -] 0.009
  Ash weight                   0.267      0.018 [+ or -] 0.002
  Ash percentage              31.847      0.594 [+ or -] 0.083
  Ca percentage               11.216      0.187 [+ or -] 0.027
  P percentage                 5.876      0.085 [+ or -] 0.017
Metatarsus mineralization
  Weight                       0.632      0.020 [+ or -] 0.003
  Length                       3.919      0.046 [+ or -] 0.007
  Ash weight                   0.175      0.010 [+ or -] 0.001
  Ash percentage              26.868      0.570 [+ or -] 0.073
  Ca percentage                9.767      0.202 [+ or -] 0.027
  P percentage                 4.733      0.113 [+ or -] 0.016

                                Slope [+ or -] SE

C riteria                        25-OH-[D.sub.3]      p value

Growth performance
  Weight gain                30.862 [+ or -] 3.726    <0.001
Femur mineralization
  Weight                      0.045 [+ or -] 0.005    <0.001
  Length                      0.118 [+ or -] 0.017    <0.001
  Ash weight                  0.028 [+ or -] 0.003    <0.001
  Ash percentage              1.356 [+ or -] 0.156    <0.001
  Ca percentage               0.485 [+ or -] 0.061    <0.001
  P percentage                0.269 [+ or -] 0.031    <0.001
Tibia mineralization
  Weight                      0.070 [+ or -] 0.008    <0.001
  Length                      0.130 [+ or -] 0.019    <0.001
  Ash weight                  0.039 [+ or -] 0.004    <0.001
  Ash percentage              1.263 [+ or -] 0.171    <0.001
  Ca percentage               0.472 [+ or -] 0.056    <0.001
  P percentage                0.214 [+ or -] 0.034    <0.001
Metatarsus mineralization
  Weight                      0.041 [+ or -] 0.006    <0.001
  Length                      0.087 [+ or -] 0.014    <0.001
  Ash weight                  0.020 [+ or -] 0.002    <0.001
  Ash percentage              1.004 [+ or -] 0.152    <0.001
  Ca percentage               0.350 [+ or -] 0.057    <0.001
  P percentage                0.187 [+ or -] 0.033    <0.001

Criteria                     [R.sup.2]    RBV [+ or -] SE

Growth performance
  Weight gain                  0.73      1.85 [+ or -] 0.23
Femur mineralization
  Weight                       0.73      1.88 [+ or -] 0.24
  Length                       0.66      1.82 [+ or -] 0.29
  Ash weight                   0.81      2.00 [+ or -] 0.20
  Ash percentage               0.73      2.03 [+ or -] 0.21
  Ca percentage                0.66      2.45 [+ or -] 0.41
  P percentage                 0.71      2.22 [+ or -] 0.29
Tibia mineralization
  Weight                       0.70      2.12 [+ or -] 0.31
  Length                       0.64      1.86 [+ or -] 0.29
  Ash weight                   0.79      2.17 [+ or -] 0.23
  Ash percentage               0.65      2.13 [+ or -] 0.32
  Ca percentage                0.68      2.52 [+ or -] 0.40
  P percentage                 0.53      2.52 [+ or -] 0.59
Metatarsus mineralization
  Weight                       0.59      2.05 [+ or -] 0.37
  Length                       0.59      1.89 [+ or -] 0.34
  Ash weight                   0.71      2.00 [+ or -] 0.27
  Ash percentage               0.65      1.76 [+ or -] 0.26
  Ca percentage                0.62      1.73 [+ or -] 0.24
  P percentage                 0.60      1.65 [+ or -] 0.27
COPYRIGHT 2016 Asian - Australasian Association of Animal Production Societies
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Article Details
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Author:Han, J.C.; Chen, G.H.; Wang, J.G.; Zhang, J.L.; Qu, H.X.; Zhang, C.M.; Yan, Y.F.; Cheng, Y.H.
Publication:Asian - Australasian Journal of Animal Sciences
Article Type:Report
Date:Jul 13, 2016
Words:6103
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