الفهرس 
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Abstract
This work was conducted to study the effects of dietary supplementation of different levels (just over the required for optimum growth) and sources (Inoragnic, Organic and Nano source) of copper on growth performance, some blood biochemical parameters, innate immune response (phagocytosis), tissue copper concentration and gut microbial diversity of Juvenile Nile tilapia. The work was conducted in two experiments:
First experiment: A total number of 720 of Juvenile Nile tilapia (Initial average weight of 4.86±0.07 g) were used in the experiment which lasted for 8 weeks. The fish were divided into 9 treatments with four replicates per each (twenty fish per tank). The experimental groups were fed the basal diets supplemented with the different copper levels as follows:
Groups
Diet Copper Sulphate
CuSO4
(mg/ kg) Tri basic copper chloride
TBCC(mg/ kg) Analyzed Cu level
(mg /kg)
1 BD* 0 0 11.2
2 BD 10 0 18.5
3 BD 75 0 75.4
4 BD 150 0 147
5 BD 300 0 287
6 BD 0 10 40.2
7 BD 0 75 83.4
8 BD 0 150 154
9 BD 0 300 300
*BD = basal diet
The obtained results can be summarized as follows:
1. Growth performance of tilapia (Body weight, weight gain, feed intake, feed conversion ratio and survival % ) was not significantly affected by the different dietary copper levels included in the diet when compared with the control group. .
2. Copper concentration in the whole body of fish was significantly increased with increasing dietary copper level (P<.0001). There was no significant difference between copper sulfate or tribasic copper chloride treatments in the whole body copper concentration of fish.
3. Whole body copper retention % was significantly (P<.0001) decreased with increasing the dietary copper level, while, was significantly affected by the copper level, form of copper used and the interaction between them.
4. Hepatosomatic index of the fish was significantly reduced (P<0.05) in the copper supplemented diet. It was the highest in the control group while the lowest in the fish fed the diet supplemented with TBCC 10 mg Cu/kg .
5. Condition factor of the fish fed diet supplemented with copper sulphate (150 and 300 mg cu/kg) was not significantly different (P>0.05) when compared with the control group. Moreover, other groups showed non-significant decrease in the condition factor when compared the control group.
6. Haematocrit (PCV %) and Alkaline phosphatase enzyme activity were significantly increased in the copper supplemented groups when compared with the control group.
7. Total and direct bilirubin showed significant (P<0.05) reduction in the copper supplemented groups when compared with the control. Also, they were significantly affected by the copper level, source and the interaction between them.
8. Cholesterol level showed non-significant increase in the copper supplemented groups except fish fed diet supplemented with 10 mg cu/kg diet (group 2) of copper sulphate which showed the lowest level when compared with the control group.
9. Non-significant increase were observed in the activities of the enzymes AST , and ALT in the copper supplemented groups except for the fish fed diet supplemented with 75 mg Cu/kg diet of TBCC showed lower activity AST and ALT.
10. Glucose was significantly affected by the copper source used (Copper sulfate or Tribasic copper chloride), also was significantly affected (P<0.05) by the interaction between the copper level and source used.
11. Total protein and the albumin showed non -significant increase in the copper supplemented groups while, globulin showed inconsistent changes in the copper sulphate supplemented groups when compared with the control group , while it was non significantly increased in the TBCC supplemented groups .
12. The mean fluorescent intensity in the copper supplemented groups was not significantly (p > 0.05) affected by the dietary copper supplementation when compared with the control group. However, it showed non-significant increase with increasing dietary copper level for the two forms of copper used .
13. Six bacterial phyla were identified from the gut faecal samples of all fish (Fusobacteria , Bacteriodetes and Proteobacteria were the highly abundant phyla ).
14. Proteobacteria was altered by the dietary supplementation of copper. It showed significant (P< 0.05) increase to be the highest in fish supplemented with TBCC 300 mg Cu/ kg diet.
15. Cetobacterium , Bacteroides, Plesiomonas and Grimontia were identified as the highly abundant genera between the different treatments .
16. Genus Grimontia was significantly (P< 0.05) increased in fish supplemented with TBCC 300 mg Cu/ kg diet when compared with other treatments.
17. Cetobacterium somerae , bacteroides sp., plesiomonas shigelloides and grimontia hollisae were idnetifed as the highly abundant species .Grimontia hollisae abundance was the highest in fish supplemented with TBCC 300 mg Cu/ kg diet when compared with other treatments.
Second experiment: A total number of 600 juvenile tilapia (Initial average weight of 3.59±0.07 g) were used in the experiment which extended for 10 weeks .The fish were randomly distributed into 5 treatments with six replicates of twenty fish per tank . The experimental groups were fed the basal diets supplemented with the different copper sources as follows:
Groups Diet Copper
supplement Supplemental copper (mg kg–1) Analyzed copper
(mg kg–1)
1 BD* Control 0 12
2 BD Copper sulphate 150 184
3 BD TBCC 150 189
4 BD Chelated-Cu 150 214
5 BD Cu- NPs 150 180
*BD = basal diet
The obtained results can be summarized as follows:
1. Growth performance parameters (Body weight, weight gain, feed intake , feed conversion ratio and survival % ) were non-significantly affected by the different copper sources when compared with the control .
2. Copper concentration in the liver and whole body of the fish was significantly (P<.0001) increased in the copper supplemented groups when compared with the control groups.
3. Inorganic copper source (copper sulfate and TBCC) supplemented groups and organic form showed the highest copper concentration in fish whole body when compared with the copper oxide nanoparticle supplemented and the control.
4. There was no significant differences in the liver copper concentration in the fish fed the diets supplemented with the copper forms, copper sulphate , TBCC and chelated copper source .
5. Copper retention % in the copper supplemented groups was significantly reduced (P<.0001) when compared with the control groups, with highest retention in the copper sulfate supplemented group.
6. There was no significant differences in the hematological parameters (WBCs, RBCs, Hb, HCT , PLT, esinophils, basophils , lympohocyte, monocytes and neutrophils ) in tilapia fed diets supplemented with different copper sources .
7. WBCs, PLT, and lymphocytes was observed to be the lowest in the fish fed the diet supplemented with the Cu-NPs.
8. Glucose level showed non-significant increase in the different copper sources supplemented groups except in the chelated- Cu group .
9. Cholesterol level was non significantly decreased in the copper supplemented groups when compared with the control group.
10. AST and ALT enzyme activities showed non-significant increase in the copper supplemented groups when compared with the control.
11. AST and AlT was non significantly (p > 0.05) increased in the fish fed diets supplemented with copper sulfate and TBCC when compared with the other fish fed on the on diets supplemented with chelated form or the copper oxide nano particles.
12. Total protein, albumin and globulin were non-significantly reduced in fish fed diets supplemented with 150 mg cu/kg diet from copper sulphate, TBCC and copper-NPs , while total protein and albumin non-significantly increased in fish fed diet supplemented with 150 mg cu/kg of chelated copper source when compared with control group .
13. Mean florescent intensity was not significantly altered (P>0.05) by dietary supplementation with different copper sources.