Dietary Supplementation of Ethanolic Lemon Peel ( Citrus limon ) Extract Enhances Growth Performance and Immune‐Antioxidant Responses in Pacific White Shrimp ( Litopenaeus vannamei )

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Abstract
Citrus ( Citrus limon ) by‐products, particularly lemon peel, are rich in bioactive compounds with known antioxidant and anti‐inflammatory properties. This study evaluated the effects of lemon peel extract (LPE) supplementation on growth performance, immune responses, and antioxidant activity in Pacific white shrimp ( Litopenaeus vannamei ). Over a 60‐day period, shrimp were fed four diets: a control diet (0%) and three diets with varying levels of LPE (0.5%, 1%, and 2%). The results showed that shrimp fed the 1% LPE diet exhibited significantly higher weight gain (WG), specific growth rate (SGR), and lower feed conversion ratio (FCR) compared to the control group ( p < 0.05). Additionally, shrimp fed 0.5% and 1% LPE showed significantly enhanced activities of antioxidant enzymes (catalase [CAT], glutathione peroxidase [GPX], superoxide dismutase [SOD]), lysozyme, total protein, and hemocyte counts, including large granular cells (GCs), semi‐granulocytes, and hyaline cells (HCs) ( p < 0.05). Total hemocyte counts (THCs) were also significantly higher in all LPE‐treated groups compared to the control ( p < 0.05). Plasma glucose levels were reduced in the 0.5% and 1% LPE treatments, though the decrease in cholesterol was not statistically significant. Moreover, the expression of integrin β , PX , and α2-M mRNA was significantly upregulated in shrimp fed the 2% LPE diet, while LGBP and proPO1 mRNA expression were higher across all LPE treatments. In conclusion, dietary supplementation with LPE, particularly at 0.5% and 1%, enhances growth performance, immune response, and antioxidant activity in L. vannamei . Therefore, LPE presents a promising and cost‐effective feed additive for shrimp aquaculture, with the potential to enhance disease resistance, support immune function, and ultimately improve shrimp health and productivity under farming conditions.

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