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Table 1.

Feed formulation with EDTA supplementation.

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Table 2.

Distribution of fish (initial weight = 30.00±1.30g) in experimental design having three stocking densities (LD, MD, HD) and their replicates at four different level of CaNa2 EDTA supplementation (E0,E1,E2,E3).

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Table 3.

Primer sequence of genes.

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Table 4.

Three density treatments (LDE, MDE, HDE) were analyzed (Mean ± SE) for total body length (cm), total body weight (g), specific growth rate (%), condition factor (%), hepatosomatic index (%), viscerosomatic index (%), FCR, and survival rate (%), with each treatment having four levels of EDTA supplementation (E0 = 0 g/kg, E1 = 5 g/kg, E2 = 10 g/kg, E3 = 15 g/kg).

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Table 5.

The proximate composition (%) of muscle samples was analyzed (mean ± SE) for each of the three density treatments (LDE, MDE, and HDE), each of which had four levels of EDTA supplementation (E0 = 0 g/kg, E1 = 5 g/kg, E2 = 10 g/kg, and E3 = 15 g/kg).

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Table 6.

Muscle samples from three density treatments (LDE, MDE, HDE) were analyzed (Mean ± SE) for amino acids (%), with each treatment having four EDTA supplementation levels (E0 = 0 g/kg, E1 = 5 g/kg, E2 = 10 g/kg, and E3 = 15 g/kg).

The results are given as milligrams of amino acid (mg/gcp) per gram of crude protein.

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Table 7.

Four EDTA supplementation levels (E0 = 0 g/kg, E1 = 5 g/kg, E2 = 10 g/kg, and E3 = 15 g/kg) were applied to each of the three density treatments (LDE, MDE, and HDE) for which the analysis of fatty acids in total lipids extracted from muscle samples was done (Mean ± SE).

The values are given as a percentage (%) of the total fats in the body.

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Table 7 Expand

Fig 1.

Three density treatments (LDE at 2.00 kg/m3, MDE at 3.50 kg/m3, HDE at 5.00 kg/m3) with four EDTA supplementation levels (E0 = 0 g/kg, E1 = 5 g/kg, E2 = 10 g/kg, and E3 = 15 g/kg) were used to assess the levels of (A) amylase, (B) lipase, and (C) protease (Mean ± SE). The three density treatments (LDE, MDE, and HDE) have significant differences (P < 0.05) indicated by superscripts in uppercase, bold, and red letters. Significant differences (P < 0.05) between the four EDTA supplementation levels within each density treatment are shown by superscripts in lowercase characters.

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Fig 1 Expand

Fig 2.

Three density treatments (LDE at 2.00 kg/m3, MDE at 3.50 kg/m3, and HDE at 5.00 kg/m3) were used to measure the cortisol level (Mean ± SE).

Each treatment had four levels of EDTA supplementation (E0 = 0 g/kg, E1 = 5 g/kg, E2 = 10 g/kg, and E3 = 15 g/kg). The three density treatments (LDE, MDE, and HDE) have significant differences (P < 0.05) indicated by superscripts in uppercase, bold, and red letters. Significant differences (P < 0.05) between the four EDTA supplementation levels within each density treatment are shown by superscripts in lowercase characters.

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Fig 2 Expand

Table 8.

Three density treatments (LDE, MDE, and HDE) were used to analyze the blood hematological parameters (Mean ± SE).

Each treatment had four levels of EDTA supplementation (E0 = 0 g/kg, E1 = 5 g/kg, E2 = 10 g/kg, and E3 = 15 g/kg).

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Table 9.

Plasma samples were subjected to three density treatments (LDE, MDE, and HDE) with four EDTA supplementation levels (E0 = 0 g/kg, E1 = 5 g/kg, E2 = 10 g/kg, and E3 = 15 g/kg) before the blood biochemistry parameters were analyzed (Mean ± SE).

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Table 10.

The assay of antioxidant biomarkers (Mean ± SE) was conducted from plasma samples from three density treatments (LDE, MDE, HDE), each with four EDTA supplementation levels (E0 = 0 g/kg, E1 = 5 g/kg, E2 = 10 g/kg, E3 = 15 g/kg).

These biomarkers included catalase (U/ml), superoxide dismutase (ng/ml), glutathione peroxidase (IU/ml), and malondialdehyde (nmol/ml).

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Fig 3.

Four EDTA supplementation doses (E0 = 0 g/kg, E1 = 5 g/kg, E2 = 10 g/kg, and E3 = 15 g/kg) were present in each of the three density treatments (LDE: 2.00 kg/m3, MDE: 3.50 kg/m3, HDE: 5.00 kg/m3) where the levels of gene expression for (A) Somatostatin 1, (B) Interleukin 1-β, and (C) POMC-α (Mean ± SE) were determined. The three density treatments (LDE, MDE, and HDE) have significant differences (P < 0.05) indicated by superscripts in uppercase, bold, and red letters. Significant differences (P < 0.05) between the four EDTA supplementation levels within each density treatment are shown by superscripts in lowercase characters.

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Fig 4.

Histological changes in gills.

Light micrographs of a paraffin section stained with eosin (10x). A(LDEO), B(LDE1), C(LDE2), D(LDE3), E(MDE0), F(MDE1), G(MDE2), H(MDE3), I(HDEO), J(HDE1), K(HDE2), L(HDE3). PL: Primary lamellae; SL: Secondary lamellae; DPL: Degeneration of primary lamellae; DSL: Degeneration of secondary lamellae; TD: Tissue debris; BC: Blood congestion; N: necrosis; EL: epithelial lifting; LF: Lamellar fusion; E: Edema; H: Hyperplasia.

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Fig 4 Expand

Table 11.

Histopathological scoring of tilapia (Oreochromis niloticus) gills in three density treatments (LDE, MDE, HDE) with each treatment having four levels of EDTA supplementation (E0 = 0 g/kg, E1 = 5 g/kg, E2 = 10 g/kg, E3 = 15 g/kg).

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