View Full Table | Close Full ViewTable 1.

Formulation of the enrichment solution

 
Mineral Chemical form Concentration Amount/embryo
Fe Bioplex Fe1 1.6 mg/mL 0.96 mg
Zn Bioplex Zn1 1 mg/mL 0.6 mg
Mn Bioplex Mn1 0.6 mg/mL 0.36 mg
Ca Organic Ca-HMB2 0.6 mg/mL 0.36 mg
Cu Bioplex Cu1 0.03 mg/mL 0.018 mg
P KH2Po4 1.4 mg/mL 0.845 mg
Maltodextrin1 4 mg/mL 2.4 mg
Vitamin A3 1,333 IU/mL 800 IU
Vitamin D3 303 IU/mL 182 IU
Vitamin E3 1.33 IU/mL 0.8 IU
1Bioplex (Alltech inc., Nicholasville, KY); chelated minerals are linked to a mixture of AA and di- and tripeptides.
2HMB = β-hydroxy-β-methylbutyrate. Sigma-Aldrich (St. Louis, MO).
3Phibro Animal Health Corp. (Teaneck, NJ).



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Composition of starter (36 h posthatch until d 10), grower (d 11 to 21), and finisher (d 22 to 54) diets fed to both groups in the experiment

 
Diet
Item Starter Grower Finisher
Ingredient, %
    Wheat 39.95 45.06 50.03
    Corn 17.80 18.40 10.80
    Dried distillers grains 4.00 9.40
    Soybean meal 27.18 19.44 14.01
    Sunflower meal 4.00 6.00 7.00
    Wheat middling 4.00
    Vegetable fat 2.64 3.02 5.06
    Limestone 1.43 1.25 1.22
    Dicalcium phosphate 1.50 1.30 1.00
    NaCl 0.22 0.20 0.18
    Vitamin–trace mineral premix1 0.90 0.80 0.75
    Hy·D2 0.05 0.05
    L-Lysine sulfate 0.11 0.23 0.32
    Methionine hydroxyl analog 0.11 0.12 0.12
    L-Thr 0.04 0.07
Calculated energy and nutrient composition, as-fed
    ME, kcal/kg 2,955 3,050 3,170
    CP, % 21.0 19.0 18.0
    Ca, % 1.16 1.00 0.90
    Total P, % 0.70 0.64 0.60
    Available P, % 0.50 0.45 0.40
    Fat, % 4.5 5.0 7.2
    Fiber, % 3.3 3.6 3.9
    Ash, % 6.19 5.57 5.24
1Provided per kilogram of grower diet (The starter diet contained 12.5% higher mineral and vitamin amounts and the finisher diet contained 6.25% lower mineral and vitamin amounts): vitamin A, 12,000 IU; vitamin D3, 5,000 IU; vitamin E, 70 IU; vitamin K, 3 mg; vitamin B1, 3 mg; vitamin B2, 7.5 mg; nicotinic acid, 50 mg; pantothenic acid, 14 mg; vitamin B6, 4 mg; folic acid, 1.5 mg; vitamin B12, 0.015 mg; biotin, 0.25 mg; Fe, 30 mg as ferrous sulfate; Zn, 100 mg as zinc oxide; Mn, 100 mg a manganous oxide; Cu, 20 mg as copper sulfate; I, 1.2 mg as potassium iodide; Co, 0.2 mg as cobalt oxide; and Se, 0.3 mg as sodium selenite.
2Hy·D = 25-hydroxycholecalciferol (DSM, Basel, Switzerland).



View Full Table | Close Full ViewTable 3.

Tibia and femur length, weight, and relative weight of the control and enriched groups in the prenatal and growing periods

 
Length, cm
Weight, g
Relative weight, mg/g BW
Control
Enriched
Control
Enriched
Control
Enriched
Sampling day1 Mean SE Mean SE Mean SE Mean SE Mean SE Mean SE
E19 Tibia 2.94 0.03 2.94 0.03 0.23 0.10 0.25 0.10 6.97 0.28 7.30 0.28
Femur 2.08 0.04 2.15 0.03 0.17 0.01 0.18 0.01 4.82 0.17 5.10 0.16
E21 Tibia 3.17 0.03 3.17 0.03 0.29 0.01 0.30 0.01 6.91 0.17 7.07 0.17
Femur 2.38 0.03 2.31 0.03 0.20 0.01 0.21 0.01 4.90 0.09 4.83 0.10
d 3 Tibia 3.54 0.02 3.54 0.02 0.50 0.01 0.48 0.01 7.65 0.21 7.08 0.21
Femur 2.58 0.03 2.54 0.03 0.32 0.01 0.30 0.01 4.87*2 0.10 4.47 0.11
d 7 Tibia 4.30 0.04 4.44* 0.04 1.02 0.03 1.04 0.03 7.08 0.19 7.17 0.19
Femur 3.23 0.05 3.21 0.05 0.71 0.02 0.73 0.02 4.88 0.14 5.00 0.14
d 14 Tibia 5.55 0.07 5.99* 0.06 2.73 0.11 3.29* 0.09 7.03 0.20 8.02* 0.18
Femur 4.07 0.07 4.25 0.06 2.05 0.08 2.35* 0.07 5.17 0.10 5.74* 0.10
d 28 Tibia 8.44 0.10 8.47 0.11 9.37 0.54 9.37 0.57 7.22 0.20 7.47 0.21
Femur 5.83 0.07 5.84 0.08 6.66 0.40 6.58 0.43 5.14 0.16 5.24 0.17
d 54 Tibia 11.88 0.14 12.00 0.18 24.36 1.40 23.52 1.76 6.87 0.17 6.63 0.21
Femur 7.99 0.12 7.94 0.12 16.81 0.89 14.64 0.89 4.80 0.19 4.45 0.19
1E19 and E21 = embryonic days.
2Means with asterisk mark a significant difference between the groups (P < 0.05); n = 8 per group on each sampling day.



View Full Table | Close Full ViewTable 4.

Bone mechanical properties in the prenatal and growing periods. stiffness, maximal load and work to fracture (WTF) of the tibia and femur1,2

 
Stiffness, N/mm
Maximal load, N
WTF, N x mm
Control
Enriched
Control
Enriched
Control
Enriched
Sampling day Mean SE Mean SE Mean SE Mean SE Mean SE Mean SE
E19 Tibia 32.26 2.15 33.19 2.15 12.11 0.81 12.94 0.81 21.68 1.81 21.44 1.57
Femur 44.42 2.57 52.20* 2.38 14.93 1.13 18.78* 1.04 12.67 1.67 19.40* 1.55
E21 Tibia 44.89 2.80 58.55* 2.80 14.28 1.16 18.66* 1.16 37.11 2.76 45.99* 2.76
Femur 35.79 2.10 44.16* 2.24 13.62 0.80 17.48* 0.86 24.98 2.55 23.99 2.73
d 3 Tibia 58.37 2.12 69.26* 2.12 22.87 0.58 22.09 0.58 54.77 1.85 53.05 1.85
Femur 80.44 2.85 89.10* 2.85 24.00 1.15 23.11 1.15 47.04 3.65 43.64 3.65
d 7 Tibia 156.5 25.7 146.9 29.5 73.7 12.6 70.6 9.9 103.5 7.2 99.7 6.7
Femur 126.4 10.3 124.7 10.3 52.1 4.5 57.5 4.5 65.6 3.1 56.5 3.1
d 14 Tibia 246.7 14.1 294.3* 14.1 139.3 7.7 185.8* 7.7 154.1 15.4 149.7 15.4
Femur 219.0 12.8 229.7 12.0 160.3 8.7 177.2 8.1 173.3 21.7 187.1 20.3
d 28 Tibia 361.2 38.7 380.6 44.7 270.1 19.3 237.8 22.3 445.8 103.0 463.5 119.0
Femur 266.1 25.1 311.4 29.0 197.8 17.9 210.1 20.7 546.3 61.7 679.5 71.3
d 54 Tibia 309.2 28.5 344.3 28.5 455.1 46.0 429.8 46.0 710.8 95.9 642.3 95.9
Femur 430.5 29.9 409.8 34.5 312.5 20.7 285.8 24.0 527.1 71.5 485.2 82.6
1E19 and E21 = embryonic days.
2Means with asterisk mark a significant difference between the groups (P < 0.05); n = 8 per group on each sampling day.



View Full Table | Close Full ViewTable 5.

Cortical bone structural properties in the prenatal and growing periods. Cortical area, medullary area, crossectional thickness, and maximal moment of inertia (Imax) of the tibia and femur

 
Cortical area, mm2
Medullary area, mm2
Crossectional thickness, mm
Imax, mm4
Control
Enriched
Control
Enriched
Control
Enriched
Control
Enriched
Sampling day1 Mean SE Mean SE Mean SE Mean SE Mean SE Mean SE Mean SE Mean SE
E19 Tibia 1.95 0.07 1.82 0.08 0.57 0.06 0.57 0.06 0.25 0.01 0.28 0.02 0.49 0.04 0.46 0.04
Femur 1.89 0.10 1.81 0.10 0.66 0.05 0.87*2 0.06 0.25 0.01 0.25 0.01 0.46 0.04 0.53 0.04
E21 Tibia 1.45 0.10 1.51 0.11 1.24* 0.08 0.98 0.08 0.27 0.02 0.31 0.02 0.49 0.04 0.44 0.05
Femur 1.80 0.10 1.98 0.09 1.11* 0.07 0.88 0.06 0.32 0.01 0.34 0.01 0.53 0.04 0.57 0.04
d 3 Tibia 1.58 0.04 1.60 0.04 1.73 0.10 1.80 0.10 0.26 0.01 0.26 0.01 0.70 0.04 0.73 0.04
Femur 1.67 0.04 1.81* 0.04 1.63 0.27 1.50 0.27 0.28 0.01 0.29 0.01 0.67 0.04 0.72 0.04
d 7 Tibia 3.30 0.15 3.58 0.14 2.26 0.08 2.22 0.08 0.43 0.02 0.44 0.02 2.38 0.18 2.44 0.18
Femur 4.55 0.14 4.60 0.14 2.24 0.08 2.16 0.08 0.53 0.01 0.53 0.01 3.97 0.18 3.89 0.18
d 14 Tibia 11.13 0.63 12.47 0.59 2.98 0.15 3.37 0.14 0.94 0.03 1.02 0.03 16.3 1.66 20.45* 1.55
Femur 13.31 0.59 14.93 0.55 4.32 0.33 4.80 0.31 0.88 0.01 0.90 0.01 23.3 2.05 29.97* 1.92
d 28 Tibia 22.32 1.27 23.15 1.36 11.11 1.26 11.67 1.35 1.21 0.07 1.21 0.07 93.7 11.2 102.8 12.0
Femur 27.87 0.01 28.97 1.16 19.72 1.48 19.02 1.71 1.07 0.06 0.98 0.07 157.9 13.7 154.1 14.6
d 54 Tibia 37.99 3.29 38.87 3.80 38.32 2.88 33.68 3.32 1.00 0.09 1.20 0.10 433.4 61.6 367.0 72.8
Femur 39.02 2.97 36.76 2.97 56.57 4.97 60.64 4.97 0.71 0.06 0.67 0.06 485.3 60.9 471.2 60.9
1E19 and E21 = embryonic days.
2Means with asterisk mark a significant difference between the groups (P < 0.05); n = 8 per group on each sampling day.



View Full Table | Close Full ViewTable 6.

Analysis of some of the different minerals in the tibia expressed in percent of the dry weight of the bone

 
Ca, %
Mn, %
P, %
Zn, %
Control
Enriched
Contro
Enriched
Control
Enriched
Control
Enriched
Sampling day1 Mean SE Mean SE Mean SE Mean SE Mean SE Mean SE Mean SE Mean SE
E19 17.46 1.23 26.83*2 2.39 0.002 0.001 0.008* 0.001 9.32 0.60 14.00* 1.23 0.01 0.002 0.02 0.004
E21 42.18 0.97 43.48 1.77 0.002 0.000 0.012* 0.001 22.14 0.53 22.66 0.92 0.06 0.004 0.06 0.003
d 3 41.64 2.65 35.09 3.83 0.002 0.000 0.006* 0.001 21.04 1.41 17.77 1.79 0.05 0.005 0.05 0.006
d 7 47.19* 2.55 39.26 1.61 0.002 0.000 0.003* 0.000 24.24* 1.32 20.57 0.87 0.05 0.003 0.06 0.010
s1E19 and E21 = embryonic days.
2Means with asterisk differ from the control (P < 0.05); n = 8 per group on each sampling day.