Increased non-specific immune activity of vaname shrimp Litopenaeus vannamei using a leaf flour mixture from Ocimum basilicum and Piper betle and their characteristic compounds

Abstract

Immunostimulants are more suitable for improving the health of shrimp than using drugs in the prevention of diseases. This study evaluated the efficacy of bioactive compounds from Ocimum basilicum and Piper betle in increasing the non-specific immune response of Litopenaeus vannamei. Samples were collected directly from a traditional market. Shrimp was fed: O. basilicum leaf flour- and P. betle leaf flour-un-supplemented diet (P1), O. basilicum leaf flour (44.84 g/kg feed)-supplemented diet (P2), P. betle leaf flour (31.57 g/kg feed)-supplemented diet (P3), and O. basilicum leaf flour (22.42 g/kg feed)- and P. betle leaf flour (16.79 g/kg feed)-supplemented diet (P4). Phytochemical screening tests were conducted using commonly used chemical reagents, including Dragendorff, Meyer, and Liebermann-Burchard reagents. Supplementation with O. basilicum and P. betle (P4) significantly enhanced the shrimp’s immune response, namely the total hemocyte counts (THC), phagocytic activity (PA), phagocytic index (PI), and prophenoloxidase (proPO). The supplemented diet (P4) was synergistically able to improve shrimp health, with the highest values for THC (7.30 x 106 cells/mL), hyaline (61.33%), PA (84%), PI (2.19), and proPO (0.91). Flavonoids and alkaloids in the feed containing leaf flour from both plants were bioactive compounds suspected to be responsible for enhancing the immune response. These findings suggest that a leaf flour combination of O. basilicum and P. betle can be a new immunostimulant alternative to improve shrimp health for disease prevention.