The primary aim of the present study is to evaluate the highest concentrations of heavy metals (HMs) in Nile catfish (Clarias gariepinus, C. gariepinus) and water samples, as well as to investigate the efficiency of pomegranate peels (PPs) (supplemented either through water or diet) in enhancing fish immunity and counteracting the toxicity of high concentrations of HMs. Water and C. gariepinus samples were collected from two private fish farms in Al Sharkia Governorate. Mercury (Hg) showed the highest concentration (0.13 ppm). The adsorption capacity of PP was evaluated by testing different concentrations, 0.3, 0.6, 0.9, 1.2, and 1.5 g L-1, wherein 1.5 g L-1 revealed the highest Hg adsorption efficiency. The results indicated that Hg concentration was decreased with increased PP concentration until 72 h. In a trial that lasted for 60 days, 240 C. gariepinus (75.12 ± 3.12 g) were randomly divided into eight equal groups with three replicates per group. The first group (CT) served as the negative control (fish fed on a basal diet). The second group (PPW) was fed on a basal diet and supplemented with 0.3 g L-1 of PP via water. The third (PPD1) and fourth (PPD2) groups received basal diets enriched with 1 and 2 g PP powder/kg diet. respectively. The fifth group (Hg) served as the positive control that was fed on a basal diet and exposed to 0.13 ppm of Hg. The sixth (Hg + PPW), seventh (Hg + PPD1), and eighth (Hg + PPD2) groups were exposed to 0.13 ppm of Hg and received the same type of treatment as in second, third, and fourth groups. Hg exposure significantly elevated aspartate aminotransferase (AST), alanine aminotransferase (ALT), urea, and creatinine levels, as well as oxidative stress biomarkers, including reduced glutathione (GSH) and malondialdehyde (MDA). Pomegranate supplementation through diet elevated the levels of red blood cells (RBCs), hemoglobin (Hb), packed cell volume (PCV), lysozyme, and anti-protease activity. Moreover, PP supplementation through water revealed minimum urea and creatinine levels, and the highest nitric oxide level. Moreover, Hg residue level in fish muscles noticeably decreased in the PP-treated groups. These results demonstrated the efficiency of PP supplementation (either through water or diet) in enhancing fish immunity and counteracting subchronic Hg toxicity.
Keywords: Antitoxic; Clarias gariepinus; Heavy metals; Immunity; Mercury; Pomegranate peel.