Table 1
Ingredients and proximate nutrient composition percentage of the control diet s (g/Kg diet, dry matter)
Ingredients %
|
|
Fish meal (65%)
|
10
|
Soybean meal (45%)
|
46.3
|
Corn gluten meal
|
3.00
|
Yellow corn
|
16.6
|
Wheat flour
|
18.5
|
Soybean oil
|
3.4
|
Vitamin mixture*
|
0.8
|
Mineral mixture**
|
0.5
|
DiCaP
|
0.6
|
Choline chloride
|
0.2
|
Stay C***
|
0.1
|
Composition (%)
|
|
Crude protein
|
32.1
|
DE (Kcal/Kg)
|
3001
|
Crude lipid
|
5.74
|
Ash
|
5.169
|
Crude fiber
|
4.11
|
Ca
|
0.798
|
P
|
0.825
|
Cu (mg/kg)
|
13.700
|
*Vitamin (g/kg premix): Thiamin HCl, 0.44; Riboflavin, 0.63; Pyridoxine HCl, 0.91; DL pantothenic acid, 1.72; Nicotinic acid, 4.58; Biotin, 0.21; Folic acid, 0.55; Inositol, 21.05; Menadione sodium bisulfite, 0.89; Vitamin A acetate, 0.68; Vitamin D3, 0.12; dL-alpha-tocoperol acetate, 12.63; Alpha-cellulose, 955.59.
**Trace mineral, copper-free (g/100 g premix): Cobalt chloride, 0.004; Furrous sulfate, 4.000; Magnesium sulfate anhydrous, 13.862; Manganous sulfate monohydrate, 0.650; Potassium iodide, 0.067; Sodium selenite, 0.010; Zinc sulfate hepahydrate, 13.193; Alpha-cellulose, 67.964.
***Stay C®, (L-ascorbyl-2-polyphosphate 35%).
Table 2
Growth performance, feed utilization and biological parameters of Nile tilapia as affected by dietary additives of bulk and Nano copper
|
Experimental treatments
|
± MSE
|
P value
|
Control
|
Bulk-Cu
(mg kg-1 diet)
|
|
Nano-Cu
(mg kg-1 diet)
|
3
|
6
|
|
3
|
6
|
Initial body weight (IBW; g fish-1)
|
9.09
|
9.08
|
9.10
|
|
9.11
|
9.12
|
0.014
|
0.2903
|
Final body weight (FBW; g fish-1)
|
41.14c
|
48.18b
|
47.47b
|
|
52.67a
|
52.10a
|
1.02
|
0.0001
|
Final body length (FBL)
|
13.03d
|
14.30b
|
14.10c
|
|
14.77a
|
14.70a
|
0.56
|
0.0001
|
Condition factor (K)
|
1.86a
|
1.65b
|
1.69b
|
|
1.64b
|
1.64b
|
0.53
|
0.0003
|
Weight gain (WG; g fish-1)
|
32.05c
|
39.10b
|
38.37b
|
|
43.56a
|
42.98a
|
1.11
|
0.0001
|
Specific growth rate (SGR; % day-1)
|
1.79c
|
2.00ab
|
1.99b
|
|
2.05a
|
2.05a
|
0.02
|
0.0001
|
Fish survival (FS %)
|
80.00b
|
93.33a
|
96.67a
|
|
100a
|
100a
|
1.17
|
0.0017
|
Feed intake (FI; g fish-1)
|
59.61d
|
68.42b
|
65.24c
|
|
71.44a
|
70.77ab
|
1.41
|
0.0001
|
Feed conversion ratio (FCR)
|
1.86a
|
1.75b
|
1.70c
|
|
1.65d
|
1.64d
|
0.54
|
0.0001
|
Protein efficiency ratio (PER)
|
1.79d
|
1.90c
|
1.96b
|
|
2.03a
|
2.02a
|
0.21
|
0.0001
|
Means followed by different small letters in the same row are significantly different (P < 0.05, one-way ANOVA)
Table 3
Histomorphometric of intestine of Nile tilapia as affected by dietary additives of bulk and Nano Copper
|
Experimental treatments
|
± MSE
|
P value
|
Control
|
Bulk-Cu
(mg kg-1 diet)
|
|
Nano- Cu
(mg kg-1 diet)
|
3
|
6
|
3
|
6
|
Anterior intestine
|
Villi Height
|
322.15c
|
479.52b
|
489.33b
|
|
685.81a
|
691.52a
|
3.69
|
0.001
|
Villi width
|
30.25c
|
33.87b
|
334.08b
|
|
32.52b
|
38.62a
|
0.562
|
0.001
|
Goblet cells number
|
27.12c
|
33.00c
|
39.90b
|
|
40.19ab
|
47.42a
|
0.236
|
0.001
|
MSR‡‡
|
8.27c
|
8.82c
|
8.35c
|
|
13.29b
|
16.17a
|
0.452
|
0.001
|
AAV††
|
45.66b
|
46.13b
|
44.46b
|
|
66.29a
|
47.95b
|
0.598
|
0.001
|
Crypt width
|
58.42b
|
62.13a
|
66.02a
|
|
62.51a
|
48.92c
|
1.89
|
0.002
|
Posterior intestine
|
Villi Height
|
186.22d
|
192.58c
|
268.59b
|
|
274.33a
|
292.69a
|
20.23
|
0.039
|
Villi width
|
52.23c
|
66.22b
|
75.89b
|
|
78.51b
|
89.26a
|
5.69
|
0.008
|
Goblet cells number
|
43.58
|
42.00
|
41.09
|
|
41.01
|
42.05
|
0.987
|
0.154
|
MSR‡‡
|
4.29a
|
5.82b
|
6.12b
|
|
9.05a
|
9.57a
|
0.878
|
0.001
|
AAV††
|
10.65c
|
12.08c
|
13.04c
|
|
15.30a
|
18.20a
|
2.56
|
0.702
|
Crypt width
|
42.03b
|
48.65b
|
59.12a
|
|
38.93c
|
35.65c
|
1.59
|
0.001
|
Means followed by different letters in the same row are significantly different (P < 0.05, one-way ANOVA)
‡‡MSR = mucosal to serosal amplification ratio
††AAV = absorption area of villous
Table 4
Hematological parameters of Nile tilapia as affected by dietary additives of bulk and Nano Copper
|
Experimental treatments
|
± MSE
|
P value
|
Control
|
Bulk-Cu
(mg kg-1 diet)
|
|
Nano-Cu
(mg kg-1 diet)
|
3
|
6
|
|
3
|
6
|
Hb (g dL-1)
|
10.11c
|
11.50b
|
12.70a
|
|
12.55a
|
13.25a
|
0.578
|
0.0318
|
Hct (%)
|
28.27d
|
36.37c
|
38.33b
|
|
36.44c
|
40.10a
|
2.919
|
0.0001
|
RBCs (×10^12 L-1)
|
2.57c
|
2.98b
|
3.14a
|
|
2.9b
|
3.23a
|
0.381
|
0.0001
|
MCV (fl)
|
110.09b
|
122.76a
|
122.82a
|
|
126.80a
|
124.25a
|
2.064
|
0.0032
|
MCH (pg)
|
39.44
|
39.63
|
40.96
|
|
44.08
|
41.66
|
1.196
|
0.4705
|
MCHC (g dL-1)
|
35.82
|
32.08
|
33.29
|
|
34.64
|
33.49
|
1.719
|
0.4531
|
WBCs (×10^9 L-1)
|
219.48b
|
170.27d
|
205.85c
|
|
216.88b
|
240.48a
|
2.562
|
0.0001
|
LYM (×10^9 L-1)
|
208.51b
|
161.76d
|
195.56c
|
|
206.04b
|
228.46a
|
2.434
|
0.0001
|
MID (×10^9 L-1)
|
10.20b
|
7.92d
|
9.57c
|
|
10.09b
|
11.18a
|
0.119
|
0.0001
|
GRAN (×10^9 L-1)
|
0.77b
|
0.60d
|
0.72c
|
|
0.76b
|
0.84a
|
0.059
|
0.0001
|
Means followed by different small letters in the same row are significantly different (P<0.05, one-way ANOVA)
*Hb = Hemoglobin; †Hct = Hematocrit; ‡RBCs = Red blood cells count; §MCV = Mean corpuscular volume; ¶MCH = Mean corpuscular hemoglobin; **MCHC = Mean corpuscular hemoglobin concentration; ††MPV = Mean platelet volume; ‡‡OCCB = Oxygen carrying capacity of blood
WBCs = White blood cells (as a total count)
LYM means lymphocyte absolute count. Lymphocytes occur in two forms: B cells,
Lymphocyte percent. Leukocyte that normally makes up about 25% of the total white blood cells count.
MID means mid-range absolute count
Mid-range percent
GRAN means Granulocyte absolute count.
Table 5
Alanine aminotransferase, aspartate aminotransferase, plasma total protein, albumin, globulin, albumin/globulin ratio and plasma lipids profiles of Nile tilapia as affected by dietary additives of bulk and Nano Copper
|
Experimental treatments
|
± MSE
|
P value
|
Control
|
Bulk-Cu
(mg kg-1 diet)
|
|
Nano-Cu
(mg kg-1 diet)
|
3
|
6
|
3
|
6
|
ALT (U L-1)
|
34.78a
|
22.36c
|
23.45b
|
|
22.08c
|
21.60c
|
0.294
|
0.0001
|
AST (U L-1)
|
93.53a
|
77.68b
|
76.27b
|
|
75.42b
|
75.12b
|
0.812
|
0.0001
|
TP (g dL-1)
|
2.57c
|
2.54c
|
2.71b
|
|
3.26a
|
2.85b
|
0.203
|
0.0140
|
Al (g dL-1)
|
0.90c
|
0.92c
|
0.98bc
|
|
1.15a
|
1.04ab
|
0.034
|
0.0052
|
Gl (g dL-1)
|
1.67c
|
1.62c
|
1.67c
|
|
2.11a
|
1.87b
|
0.197
|
0.0137
|
Al/Gl ratio
|
0.57b
|
0.58b
|
0.65a
|
|
0.54b
|
0.54b
|
0.044
|
0.8014
|
TC (mg dL-1)
|
179.53a
|
158.10b
|
157.86b
|
|
154.10b
|
158.42b
|
2.722
|
0.0011
|
TG (mg dL-1)
|
180.14a
|
138.27b
|
125.18c
|
|
135.20b
|
132.63bc
|
2.548
|
0.0001
|
HDL-C (mg dL-1)
|
35.43c
|
44.91b
|
46.01a
|
|
44.26b
|
46.53a
|
0.028
|
0.0001
|
LDL-C (mg dL-1)
|
108.08a
|
85.53b
|
86.81b
|
|
82.80b
|
85.37b
|
2.940
|
0.0016
|
VLDL-C (mg dL-1)
|
36.03a
|
27.65b
|
25.04c
|
|
27.04b
|
26.53bc
|
0.509
|
0.0001
|
|
|
|
|
|
|
|
|
|
|
|
Means followed by different small letters in the same row are significantly different (P<0.05, one-way ANOVA)
Where: *ALT = Alanine aminotransferase; †AST = Aspartate aminotransferase.
*TP = Total protein; †Al = Albumin; ‡Gl = Globulin; §Al/Gl ratio = Albumin/globulin ratio. *TC = Total cholesterol; †TG = Triglycerides; ‡HDL-C = High-density lipoprotein cholesterol; §LDL-C = Low-density lipoprotein cholesterol; ¶VLDL-C = Very low-density lipoprotein cholesterol.
Table 6
Lysozyme, Catalase, Superoxide dismutase, Glutathione peroxidase, Glutathione, Glutathione S-transferases, glutathione-disulfide reductase and Malondialdehyde of Nile tilapia as affected by dietary additives of bulk and Nano Copper
|
Experimental treatments
|
± MSE
|
P value
|
Control
|
Bulk-Cu
(mg kg-1 diet)
|
|
Nano-Cu
(mg kg-1 diet)
|
0.3
|
0.6
|
0.3
|
0.6
|
LZM (ng mL-1)
|
0.70c
|
0.80b
|
0.81b
|
|
0.84a
|
0.85a
|
0.055
|
0.0001
|
CAT (ng mL-1)
|
50.50d
|
51.10d
|
58.30b
|
|
52.80c
|
63.43a
|
1.389
|
0.0001
|
SOD (U mL-1)
|
25.06c
|
29.15b
|
34.26a
|
|
28.23b
|
33.86a
|
0.395
|
0.0001
|
GPx (ng mL-1)
|
25.94d
|
27.94c
|
31.08b
|
|
26.10d
|
32.42a
|
0.156
|
0.0001
|
MDA (nmol mL-1)
|
73.95a
|
69.20b
|
69.17b
|
|
66.64b
|
62.74c
|
0.806
|
0.0001
|
Means followed by different small letters in the same row are significantly different (P<0.05, one-way ANOVA)
Where: *LZM = Lysozyme; †CAT = Catalase; ‡SOD = Superoxide dismutase; §GPx = Glutathione peroxidase; ¶GSH = Glutathione; **GST = Glutathione S-transferases; ††GSR = glutathione-disulfide reductase; ‡‡MDA = Malondialdehyde.
Table 7
Proximate analysis of flesh of Nile tilapia as affected by dietary additives of bulk-Se and Nano copper
|
Experimental treatments
|
± MSE
|
P value
|
Control
|
Bulk-Cu
(mg kg-1 diet)
|
|
Nano-Cu
(mg kg-1 diet)
|
3
|
6
|
3
|
6
|
Chemical composition of fish flesh
|
Moisture (%)
|
74.80a
|
74.61ab
|
74.10bc
|
|
73.72c
|
73.67c
|
0.185
|
0.0076
|
Protein (%)
|
18.79d
|
19.41c
|
19.89b
|
|
20.18a
|
20.37a
|
0.048
|
0.0001
|
Fat (%)
|
2.93a
|
2.50b
|
2.31c
|
|
2.20cd
|
2.05d
|
0.015
|
0.0001
|
Ash (%)
|
2.01e
|
2.25d
|
2.43c
|
|
2.59b
|
2.73a
|
0.032
|
0.0001
|
Cu (mg kg-1)
|
0.49d
|
0.51bc
|
0.63b
|
|
1.18a
|
1.23a
|
0.014
|
0.0001
|
Means followed by different small letters in the same row are significantly different (P<0.05, one-way ANOVA)