Table 1 Effects of supplementation of calcidiol, DHA-rich microalgal biomass or oil, and EPA-rich microalgal biomass on body weight, feed intake, and gain: feed ratio in broiler chickens in Experiments 1 and 2 (0-6 wk)
Experiment 1
|
Parameters
|
BD
|
BD+Cal
|
BD+DHA
|
BD+Cal+DHA
|
SEM
|
P-value
|
|
BW 0 day (g/chick)
|
41.85
|
41.77
|
41.80
|
41.65
|
0.17
|
0.85
|
|
BW 0-6 wk (g/chick)
|
2913.20a
|
3232.62b
|
3084.55ab
|
3160.82ab
|
51.20
|
<0.01
|
|
BWG 0-6 wk (g/chick)
|
2871.34a
|
3190.84b
|
3042.75ab
|
3119.17ab
|
57.25
|
<0.01
|
|
FI 0-6 wk (g/chick)
|
4860.10a
|
5050.36ab
|
5240.20ab
|
5450.56b
|
115.4
|
<0.01
|
|
Gain: Feed
|
0.59a
|
0.63b
|
0.58a
|
0.57a
|
0.008
|
<0.01
|
|
Experiment 2
|
|
BD
|
BD+DHA
|
BD+DHA+EPA
|
BD+DHA
+Cal
|
BD+DHA+EPA+Cal
|
SEM
|
P-value
|
BW 0 day (g/chick)
|
42.25
|
42.32
|
42.39
|
42.43
|
42.36
|
0.07
|
0.64
|
BW 0-6 wk (g/chick)
|
2522.33a
|
2598.92ab
|
2774.17b
|
2533.25ab
|
2592.92ab
|
62.82
|
0.04
|
BWG 0-6 wk (g/chick)
|
2480.08a
|
2556.60ab
|
2731.78b
|
2490.82ab
|
2550.56ab
|
62.84
|
0.04
|
FI 0-6 wk (g/chick)
|
3875.52a
|
3867.97a
|
4413.40b
|
4075.90ab
|
4474.10b
|
121.8
|
<0.01
|
Gain: Feed
|
0.64ab
|
0.66b
|
0.62ab
|
0.61ab
|
0.57a
|
0.02
|
0.02
|
BW: Body weight; BWG: Body weight gain; FI: Feed intake.
Experiment 1; BD = Corn-soybean basal diet; BD+Cal = BD + 10000 IU calcidiol/Kg of diet; BD+DHA = BD + 1% microalgal biomass; BD+Cal+DHA = BD+Cal + 1% microalgal biomass.
Experiment 2; 0-3 Wk: BD = Corn-soybean basal diet; BD+DHA= BD + 1.5 g/kg DHA oil; BD+DHA+EPA = BD+DHA + 0.3 g/kg Nannochloropsis sp CO18; BD+DHA+Cal = BD+DHA + 6000 IU/kg calcidiol; BD+DHA+EPA+Cal = BD+DHA+EPA + 6000 IU/kg calcidiol.
4-6 Wk: BD = Corn-soybean basal diet; BD+DHA= BD + 3.0 g/kg DHA oil; BD+DHA+EPA = BD+DHA + 0.6 g/kg Nannochloropsis sp CO18; BD+DHA+Cal = BD+DHA + 12000 IU/kg calcidiol; BD+DHA+EPA+Cal = BD+DHA+EPA + 12000 IU/kg calcidiol.
Means bearing the different superscripts (a, b) in a row differ significantly (P <0.05).
Data are expressed as means (n = 6 cages and 6 birds/cage) and were analyzed by one-way ANOVA.
Table 2 Effects of supplementation of calcidiol and DHA-rich microalgal biomass on plasma lipid profile of broiler chickens in Experiment 1
|
BD
|
BD+Cal
|
BD+DHA
|
BD+Cal+DHA
|
SEM
|
P-value
|
Wk 3
|
TG (mg/dL)
|
22.48
|
22.21
|
25.27
|
28.07
|
3.24
|
0.65
|
TC (mg/dL)
|
133.17
|
136.11
|
139.29
|
129.68
|
5.03
|
0.62
|
NEFA (µmol/mL)
|
0.21
|
0.18
|
0.17
|
0.16
|
0.02
|
0.27
|
Wk 6
|
TG (mg/dL)
|
20.26
|
23.49
|
21.84
|
22.52
|
1.97
|
0.55
|
TC (mg/dL)
|
101.16b
|
77.96a
|
97.76b
|
100.53b
|
4.42
|
<0.01
|
NEFA (µmol/mL)
|
0.24
|
0.24
|
0.24
|
0.28
|
0.03
|
0.69
|
NEFA: Non-esterified fatty acid; TC: Total cholesterol; TG: Triglyceride.
BD = Corn-soybean basal diet; BD+Cal = BD + 10000 IU calcidiol/kg of diet; BD+DHA = BD + 1% microalgal biomass; BD+Cal+DHA = BD+Cal + 1% microalgal biomass.
Means bearing the different superscripts (a, b) in a row differ significantly (P <0.05).
Values are expressed as means of 6 birds/treatment and data were analyzed using one-way ANOVA.
Table 3 Effects of supplementation of DHA-rich microalgal oil, EPA-rich microalgal biomass, and calcidiol on plasma and tissue lipid profile of broiler chickens at wk 3 in Experiment 2
|
BD
|
BD+DHA
|
BD+DHA
+EPA
|
BD+DHA
+Cal
|
BD+DHA
+EPA+Cal
|
SEM
|
P-value
|
Plasma
|
PL (mg/dL)
|
102.28
|
90.22
|
91.54
|
100.57
|
104.44
|
6.26
|
0.48
|
TG (mg/dL)
|
31.64
|
25.51
|
33.74
|
25.23
|
34.95
|
2.88
|
0.08
|
TC (mg/dL)
|
100.3
|
103.29
|
90.64
|
105.5
|
106.76
|
2.50
|
0.41
|
NEFA (µmol/mL)
|
219.42
|
203.65
|
202.14
|
229.52
|
212.39
|
15.60
|
0.72
|
Liver
|
PL (mg/g tissue)
|
16.72
|
14.74
|
15.20
|
16.14
|
15.80
|
2.42
|
0.16
|
TG (mg/g protein)
|
66.76
|
70.12
|
64.81
|
65.08
|
70.04
|
5.17
|
0.95
|
TC (mg/g protein)
|
15.28
|
14.12
|
15.02
|
14.80
|
14.61
|
1.14
|
0.81
|
NEFA (µmol/g protein)
|
45.34
|
46.80
|
41.12
|
41.95
|
43.44
|
3.80
|
0.65
|
Breast
|
PL (mg/g tissue)
|
6.01
|
4.16
|
6.12
|
5.76
|
6.09
|
0.44
|
0.06
|
TG (mg/g protein)
|
21.44
|
21.16
|
19.30
|
21.05
|
20.82
|
1.36
|
0.73
|
TC (mg/g protein)
|
6.36
|
6.10
|
6.53
|
6.69
|
6.80
|
1.24
|
0.80
|
NEFA (µmol/g protein)
|
5.27
|
5.12
|
4.85
|
4.96
|
4.79
|
0.70
|
0.56
|
Thigh
|
PL (mg/g tissue)
|
3.75
|
3.25
|
3.43
|
3.64
|
4.34
|
0.52
|
0.64
|
TG (mg/g protein)
|
23.50
|
22.14
|
21.60
|
23.44
|
22.40
|
1.22
|
0.87
|
TC (mg/g protein)
|
4.56
|
4.60
|
4.53
|
4.37
|
4.32
|
0.51
|
0.64
|
NEFA (µmol/g protein)
|
5.50
|
5.33
|
5.06
|
5.53
|
5.24
|
0.83
|
0.71
|
BD = Corn-soybean basal diet; BD+DHA= BD + 1.5 g/kg DHA oil; BD+DHA+EPA= BD+DHA + 0.3 g/kg Nannochloropsis sp CO18; BD+DHA+Cal = BD+DHA + 6000 IU/kg calcidiol; BD+DHA+EPA+Cal = BD+DHA+EPA + 6000 IU/kg calcidiol.
Means bearing no superscripts did not differ significantly (P >0.05).
Values are expressed as means of 6 birds/treatment and data were analyzed using one-way ANOVA.
Table 4 Effects of supplementation of DHA-rich microalgal oil, EPA-rich microalgal biomass, and calcidiol on plasma and tissue lipid profile of broiler chickens at wk 6 in Experiment 2
|
BD
|
BD+DHA
|
BD+DHA
+EPA
|
BD+DHA
+Cal
|
BD+DHA
+EPA+Cal
|
SEM
|
P-value
|
Plasma
|
PL (mg/dL)
|
78.92
|
61.70
|
67.09
|
76.26
|
70.74
|
4.50
|
0.08
|
TG (mg/dL)
|
40.37b
|
27.84ab
|
18.72a
|
39.23b
|
28.77ab
|
3.76
|
<0.01
|
TC (mg/dL)
|
101.78b
|
89.57ab
|
83.76a
|
90.29ab
|
79.84a
|
3.53
|
<0.01
|
NEFA (µmol/mL)
|
87.31b
|
62.01a
|
76.64ab
|
85.62b
|
78.77ab
|
4.21
|
<0.01
|
Liver
|
PL (mg/g tissue)
|
14.60
|
12.31
|
12.48
|
14.66
|
13.65
|
1.74
|
0.22
|
TG (mg/g protein)
|
71.40b
|
68.16b
|
58.72a
|
70.85b
|
66.10ab
|
6.12
|
<0.01
|
TC (mg/g protein)
|
14.70
|
13.30
|
13.55
|
14.06
|
13.57
|
1.37
|
0.95
|
NEFA (µmol/g protein)
|
45.98
|
41.51
|
34.05
|
42.14
|
38.82
|
3.10
|
0.15
|
Breast
|
PL (mg/g tissue)
|
3.60
|
2.89
|
3.09
|
3.38
|
2.76
|
0.19
|
0.32
|
TG (mg/g protein)
|
23.77b
|
22.92b
|
18.07a
|
23.06b
|
22.88b
|
2.11
|
0.04
|
TC (mg/g protein)
|
5.82
|
4.61
|
4.69
|
5.41
|
5.59
|
0.30
|
0.05
|
NEFA (µmol/g protein)
|
5.08b
|
4.67ab
|
3.78a
|
4.50ab
|
4.44ab
|
0.91
|
0.03
|
Thigh
|
PL (mg/g tissue)
|
3.34
|
2.47
|
2.92
|
2.83
|
2.56
|
0.32
|
0.12
|
TG (mg/g protein)
|
26.62b
|
24.47ab
|
21.52a
|
24.16ab
|
23.96ab
|
2.16
|
0.01
|
TC (mg/g protein)
|
4.00
|
3.93
|
3.63
|
3.88
|
3.78
|
0.11
|
0.21
|
NEFA (µmol/g protein)
|
5.18
|
4.99
|
4.58
|
5.07
|
4.53
|
1.01
|
0.90
|
BD = Corn-soybean basal diet; BD+DHA= BD + 3.0 g/kg DHA oil; BD+DHA+EPA = BD+DHA + 0.6 g/kg Nannochloropsis sp CO18; BD+DHA+Cal = BD+DHA + 12000 IU/kg calcidiol; BD+DHA+EPA+Cal = BD+DHA+EPA + 12000 IU/kg calcidiol.
Means bearing the different superscripts (a, b) in a row differ significantly (P <0.05).
Values are expressed as means of 6 birds/treatment and data were analyzed using one-way ANOVA.
Table 5 Effects of supplementation of calcidiol and DHA-rich microalgal biomass on tibia bone properties of broiler chickens at wk 6 in Experiment 1
|
BD
|
BD+Cal
|
BD+DHA
|
BD+Cal+DHA
|
SEM
|
P-value
|
Extension at Maximum Load (mm)
|
2.13
|
2.12
|
2.06
|
2.21
|
0.10
|
0.79
|
Energy at Maximum Load (J)
|
0.47a
|
0.56b
|
0.44a
|
0.49ab
|
0.04
|
0.02
|
Maximum Slope (mm/N)
|
0.09
|
0.05
|
0.05
|
0.04
|
0.01
|
0.06
|
Maximum Load (N)
|
500.19
|
515.46
|
514.46
|
519.89
|
31.48
|
0.97
|
Maximum Extension (mm)
|
2.37
|
2.49
|
2.44
|
2.30
|
0.17
|
0.14
|
BD = Corn-soybean basal diet; BD+Cal = BD + 10000 IU calcidiol/kg of diet; BD+DHA = BD + 1% microalgal biomass; BD+Cal+DHA = BD+Cal + 1% microalgal biomass.
Means bearing the different superscripts (a, b) in a row differ significantly (P <0.05).
Values are expressed as means of 6 birds/treatment and data were analyzed using one-way ANOVA.
Table 6 Effects of supplementation of DHA-rich microalgal oil, EPA-rich microalgal biomass, and calcidiol on tibia bone properties of broiler chickens at wk 6 in Experiment 2
|
BD
|
BD+DHA
|
BD+DHA
+EPA
|
BD+DHA
+Cal
|
BD+DHA
+EPA+Cal
|
SEM
|
P-value
|
Total BMD (g/cm3)
|
0.27ab
|
0.26a
|
0.29ab
|
0.31b
|
0.30ab
|
0.01
|
0.04
|
Total Bone Volume (%)
|
34.09a
|
33.75a
|
37.40ab
|
38.21b
|
36.17ab
|
1.50
|
0.04
|
Cortical BMD (g/cm3)
|
0.60
|
0.58
|
0.61
|
0.66
|
0.65
|
0.03
|
0.38
|
Cortical Bone Volume (%)
|
99.999
|
99.999
|
100.000
|
99.992
|
99.999
|
0.0014
|
0.20
|
Trabecular
BMD (g/cm3)
|
0.10
|
0.10
|
0.12
|
0.10
|
0.10
|
0.02
|
0.94
|
Trabecular Bone Volume (%)
|
9.97
|
10.11
|
11.36
|
10.40
|
9.99
|
1.44
|
0.97
|
Trabecular Thickness (mm)
|
0.14
|
0.14
|
0.15
|
0.16
|
0.15
|
0.01
|
0.19
|
Trabecular Separation (mm)
|
1.20
|
1.31
|
1.22
|
1.24
|
1.36
|
0.11
|
0.52
|
Degree of Anisotropy
|
1.70
|
1.72
|
1.65
|
1.66
|
1.63
|
0.05
|
0.72
|
BD = Corn-soybean basal diet; BD+DHA= BD + 3.0 g/kg DHA oil; BD+DHA+EPA = BD+DHA + 0.6 g/kg Nannochloropsis sp CO18; BD+DHA+Cal = BD+DHA + 12000 IU/kg calcidiol; BD+DHA+EPA+Cal = BD+DHA+EPA + 12000 IU/kg calcidiol.
BMD = Bone mineral density.
Means bearing the different superscripts (a, b) in a row differ significantly (P <0.05).
Values are expressed as means of 6 birds/treatment and data were analyzed using one-way ANOVA.