الفهرس 
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Abstract
The present study was designed to investigate the affect of adding some phytobiotics (Fenugreek & Echinacea) and some microbial biomass (Saccharomyces cervisea & Spirulina platensis) to Nile Tilapia fish diets without or with vitamin C and E. Growth performance parameters including body weight development, body weight gain, feed intake (FI), feed conversion ratio (FCR), feed efficiency ratio (FER), protein efficiency ratio (PER), specific growth rate (SGR), fish length and fulton condition factor (FC) were recorded and calculated. However, serum biochemical parameters, serum lipid profile, serum antioxidant activity and gene expression of Nile Tilapia fish were estimated.
A total number of 330 of apparently healthy Nile Tilapia fish (Oreochromis niloticuss) were procured from a private fish farm at Cairo Governorate and transported alive to the Aquatic Animal Medicine Unit (wet lab) in plastic tank fitted with air pumps. Fish were maintained in tanks and acclimated for two weeks prior to the beginning of the experiment and fed on basal control diet. Nile tilapia fingerlings (13.0±1.0 g/fish) were randomly assigned into 10 experimental groups each with three replicates (n= 33 fish/group, 11 fish/replicate). Tested fish were stocked in 10 clean concrete ponds (220* 140*110 cm) and three cages made from plastic mesh materials of 1 mm mesh placed in each concrete pond represents three replicates for each group.
Five isoenergetic experimental diets were formulated to contain 32 % crude protein and 2872 kcal DE/kg (D1, D2, D3, D4 and D5) which fed to fish in the first 14 days of the experiment at the rate of 5% of body weight. Other
five isoenergetic experimental diets containing 28.0% crude protein and 2875 kcal DE/kg (D6, D7, D8, D9 and D10) were formulated and fed to fish at the rate of 4% of body weight at the second feeding phase (15 – 35 day). At the third feeding phase, fish fed on the same diets at 3% of body biomass. All experimental diets were formulated to meet the nutrient requirement of tilapia fish according to NRC (2011). In the first group, fish were fed on basal control diets (D1 & D6) without any additives. This group assigned as a control which the other treated groups was compared. Tilapia fish in the second experimental group fed on the same basal control diets (D1 & D6) supplemented with extra vitamin C+E at the rates of 500 and 200 mg|kg diet respectively. Nile tilapia fish in the third and fourth groups fed on second and seventh experimental diets (D2 & D7) containing 1% dried fenugreek seeds powder without or with extra 500 + 200 mg vitamin C and E/kg diet respectively. Fish in the fifth and sixth groups fed on third and eighth experimental diets (D3 &D8) containing Echinacea pruprea at the level of 5g|kg feed without or with extra vitamin C+E. In the seventh and eighth groups, fish fed on diets D4 & D9 containing 1% dried yeast (Saccharomyces cervisea) without or with Extra vitamin C + E. Nile Tilapia fish fingerlings in the ninth and tenth groups fed on fifth and tenth experimental diets (D5 & D10) supplemented with 1% Spirulina platensis without or with extra vitamin C + E (500 and 200 mg|kg). The feed quantity was readjusted in each feeding phase on the basis of the actual biomass of the fish in each group. Fish in each pond were fed by hand to apparent satiation twice daily (8.00 and 17.00) throughout the experimental period.
Experimental period was extended for 60 days duration in three feeding phases and growth performance parameters including body weight development, gain, and feed intake were recorded while, feed conversion indices, FER, PER, SGR, FC were calculated. Blood samples were collected randomly from the caudal vein of 5 fish from each group at the end of experiment. Serum biochemical parameters including total serum protein and its fractions, lipid profile, MDA, SOD, and GPx levels were determined. Pool of liver, spleen and anterior kidney were collected from nine tilapia fish. Samples were preserved in RNAlater and stored at −80˚C until RNA extraction.
The following results were obtained:
1. No mortality was recorded during the feeding period intervals in all treated groups including extra vitamin E+C, fenugreek seeds, Echinacea, Saccharomyces cervisea and Spirulina platensis supplemented groups and survival rate was enclosed to 100%.
2. Addition of extra vitamin E and C to Nile tilapia fish diets had positive effect on survival rate, growth performance including BW, BW gain feed conversion, FER, PER, specific growth rate (SGR) and Fulton condition factor. Adding extra vitamin E+C to Nile tilapia diet insignificantly (p<0.05) increased BW and WG of tilapia fish in the second group (98.40±14.36, 83.23±14.06 g) in comparison with control (94.80±11.67, 79.70±11.40 g).
3. Adding Fenugreek seeds powder to Nile tilapia diets increased significantly (p<0.05) body weight and body weight gain by about 15.79 &
18.0% in the fourth group and 14.8 &16.5 % in the third group than control at the end of the experiment (60 days).
4. Fish in the fourth group achieved best feed conversion, feed efficiency ratio, protein efficiency ratio, SGR and Fulton condition factor (1.29±0.20, 0.87± 0.15, 3.06±0.51, 3.32±0.23 and 697.07±85.38, respectively) in comparison with control (1.49±0.20, 0.70±0.09, 2.46±0.35, 3.04±0.17 and 566.13±59.04, respectively).
5. During the whole experimental period (60 days), fish in group 6 fed on diet supplemented with 0.5% Echinacea purpurea with extra vitamin E+C had significant higher body weight and weight gain (107.67±19.06, 92.03±18.82 g) than control group (94.80±11.67, 79.70±11.40 g).
6. Best feed conversion, feed efficiency ratio, protein efficiency ratio, SGR and Fulton condition factor were recorded in sixth group (1.34±0.26, 0.81± 0.16, 2.85± 0.59, 3.17± 0.27 and 656.10±105.96) in comparison with control one (1.49± 0.20, 0.70±0.09, 2.46±0.35, 3.04±0.17 and 566.13±59.04) during the whole experimental period.
7. At the end of the experiment, final weight and weight gain of fish were significantly increased in yeast supplemented groups without or with extra vitamin E+C (groups 7&8) by about 7.52, 8.19% and 8.58, 10.16 % respectively than control.
8. For the whole experimental period, fish in eighth group fed diet containing 1% Saccharomyces cervisea with vitamin E & C recorded lower FCR and higher FER, PER and SGR (1.33±0.17, 0.78±0.11, 2.75± 0.40 and 3.18±0.15 ) in comparison with fish in the seventh group without vitamin E & C (1.37±0.17, 0.76±0.11, 2.66±0.38, 3.03±0.22).
9. Fish fed on diet containing 1.0 % S. platensis with extra vitamins in group 10 recorded higher BW and BWG (104.80±11.38, 89.43±11.26 g) than those fed the same diet without vitamin E & C (101.37±12.23, 85.80±12.16 g) in comparison with control (94.80±11.67, 79.70±11.40 g).
10. Nile tilapia fish in the tenth group recorded lower FCR and higher FER, PER, SGR and Fulton condition factor (1.27±0.17, 0.82±0.10, 2.87±0.36, 3.18±0.18 and 633.27±59.49) in comparison with fish in the ninth group without vitamin E & C (1.32±0.17, 0.79±0.11, 2.76±0.39, 3.10±0.19 and
608.00±61.93).
11. Inclusion of phytobiotics or microbial biomass in tilapia fish diets without or with vitamins E+C increased significantly (P<0.05) the total serum protein, globulin values in all treated groups and fish in the sixth group recorded highest total serum protein (5.34± 0.09 g/dl) and globulin (3.77±
0.05 g/dl).
12. Feeding fish on fenugreek, Echinacea, Saccharomyces cervisea and Spirulina platensis supplemented diets showed a significant decrease in the cholesterol, triglycerides, LDL and the lowest serum cholesterol, triglycerides and LDL values was recorded in fish in the tenth group fed on diet containing 1% spirulina platensis with extra vitamin E+ C (144.77
±6.84, 92.30±4.82 and 34.03±5.19 mg/dl).
13. Fenugreek, Echinacea, Saccharomyces cervisea and Spirulina platensis supplementation without or with extra vitamins decreased significantly (P<0.05) malondialdehyde (MDA) formation, while activity of serum superoxide dismutase (SOD) and glutathione peroxidase (GPx) enzymes significantly (p<0.05) increased in all treated groups compared with control. Fish in the tenth group recorded the lowest level of
malondialdehyde (MDH) formation (14.73±2.25 µmol) and highest concentration of serum superoxide dismutase (SOD) and glutathione peroxidase (GPx) enzymes (447.67± 16.41 and 36.97± 1.71U/ml).
14. The mRNA level of IL-1β, catalase, SOD2, IL-10, and IGF-1 genes were significantly (P<0.05) increased in fish of all treated groups compared with the control one. However, the transcription of IFN-γ was insignificantly increased in the second group. Addition of fenugreek, Echinacea, Saccharomyces cervisea or Spirulina platensis to tilapia fish diets without or with vitamin C and E resulted in significant up-regulation of all the tested genes.