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Fish and Shellfish Immunology
ELsevier
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Abstract: |
Two experiments were conducted in this study, using 250 Oreochromis niloticus (O. niloticus) (average weight
30.28 ± 0.27 g). The first experiment was conducted to investigate the 96-h lethal concentration 50 (LC50) of
copper chloride (CuCl2) using the probit analysis, seventy fish was divided into seven different concentration of
CuCl2 (0, 22, 23, 24, 25, 26, and 27 mg/L), the accurate Cu concentrations were (1.23, 5.36, 6.02, 6.98, 7.05,
7.93, 8.12 mg/L Cu). The second experiment was conducted for investigating the effect of dietary supplementation
with thyme (Thymus vulgaris, T. vulgaris) and sweet basil (Ocimum basilicum, O. basilicum) essential oils
(TEO and BEO respectively) against sub-lethal Cu exposure (1/10 96-h LC50 of CuCl2). About 180 fish was
divided into six groups in triplicate (10 fish/replicate, 30 fish/group). Group 1 (C) was kept as a control group
with no Cu exposure and was fed the control basal diet. Group 2 (C–Cu) was fed the control basal diet and
simultaneously exposed to 1/10 of the 96 h LC50 of CuCl2 (2.574 mg/L) as a sub-lethal concentration of Cu,
where the realistic Cu concentration was 3.976 mg/L. Group 3 (TEO) and group 4 (BEO) were fed the diets
fortified with 1%TEO and BEO, respectively without exposure to Cu. Group 5 (TEO-Cu) and group 6 (BEO-Cu)
were fed the diets fortified with 1%TEO and 1%BEO, respectively, and simultaneously exposed to 1/10 of the 96
h LC50 of CuCl2 (2.574 mg/L). The growth and behavioral performance, immunological response and its related
gene expression, antioxidant status, stress biomarker indicators, apoptosis biomarkers, and histopathological
alteration were investigated. The results of the first experiment showed that the 96-h LC50 of CuCl2 in O. niloticus
was 25.740 mg/L with lower and upper confidence limits of 25.152 and 26.356 mg/L, respectively. The results of
the second experiment showed that sub-lethal Cu exposure induced growth retardation (lowered final body
weight, total weight gain, and specific growth rate %), behavioral abnormalities (slower swimming activity and
feeding performance), immunosuppression (lowered nitric oxide, complement-3, lysozyme, total proteins, albumin,
and globulin), and lowering the hepatic antioxidant functions (higher MDA, and lower SOD, CAT, and
GPx) in the exposed fish. Furthermore, alteration in the immune-related genes expression (down-regulation of IL-
10 and TGF-β and up-regulation of IL-1β, IL-6, IL-8, and TRL-4), hepato-renal dysfunction (elevated ALT, AST,
urea, and creatinine), and high levels of serum stress indicators (cortisol and glucose) were markedly evident.
sub-lethal Cu toxicity induced significant up-regulation of apoptosis biomarkers involving, nuclear factor-κβ (NF-
κβ), Bcl-2 Associated X-protein (BAX), meanwhile, the expression of B-cell lymphoma 2 (BCL2)
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