Effects of dietary walnut (Juglans regia) leaves extract on immunity, gene expression responses, and disease resistance in Oreochromis niloticus

The dietary effects of walnut leaf extract (WLE) on the growth, immunity, and resistance of Oreochromis niloticus to bacterial infection have been investigated. Five diets were prepared with various WLE doses of 0, 250, 500, 750, and 1000 mg/kg, termed as Con (control), WLE250, WLE500, WLE750, and WLE1000, respectively. Fish (11.67 ± 0.21 g) were fed these diets for 60 days and then challenged with Plesiomonas shigelloides. Before the challenge, it was observed that dietary WLE did not significantly affect the growth, blood proteins (globulin, albumin, and total protein), and liver function enzymes (ALT and AST) activities. The WLE250 group significantly increased serum SOD and CAT activities more than other groups. The serum immunological indices (lysozyme and myeloperoxidase activities) and hematological parameters (phagocytic activity %, phagocytic index, respiratory burst activity, and potential activity) were significantly increased in the WLE groups compared with the Con group. The expression of IgM heavy chain, IL-1β, and IL-8 genes were significantly upregulated in all WLE-supplemented groups in comparison with the Con group. The fish survival rates (SR; %) post challenge in the Con, WLE250, WLE500, WLE750 and WLE1000 groups were 40.0%, 49.3%, 86.7%, 73.3%, and 70.7%, respectively. The Kaplan-Meier survivorship curves illustrated that the highest SR% was found in the WLE500 group (86.7%) amongst the other groups. Accordingly, we can suggest that feeding O. niloticus with a diet supplied with WLE at a dose rate of 500 mg/kg over 60 days could enrich haemato-immune responses and increase the fish survival against the challenge with P. shigelloides. These results recommend using WLE as a herbal dietary supplement to substitute antibiotic use in aquafeed.