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Here we attempted to synthesize β-1,3-glucan binding protein-based silver nanoparticles (Phβ-GBP-AgNPs) and evaluate its wound healing and disease resistance prompting ability in Oreochromis mossambicus. Using a column chromatography technique, an immune molecule, Phβ-GBP was purified from the haemolymph of rice field crab, Paratelphusa hydrodromus. Phβ-GBP-AgNPs were synthesized and described through SDS-PAGE, UV-vis spectroscopy, HR-TEM, XRD and FTIR analysis. HR-TEM revealed that the synthesized Phβ-GBP-AgNPs were spherical with a 20-40 nm size range and the particles were not aggregated. Wound and infection were experimentally generated in O. mossambicus and treated with Phβ-GBP, chem-AgNPs and Phβ-GBP-AgNPs for 20 days. The immune parameters (peroxidase, lysozyme and protease) and antioxidant enzymes (SOD and catalase) were examined in the serum of experimental fish. Phβ-GBP-AgNPs elevated the immune and antioxidant enzymes during the healing process and enhanced the wound healing percentage in fish than Phβ-GBP and chem-AgNPs. The immune parameters and antioxidant enzymes were declined in the serum of fish (treated with Phβ-GBP-AgNPs) after the mid-period of wound healing. Compared to others, relative percentage survival was increased in experimentally wounded and infected fish treated with Phβ-GBP-AgNPs against Aeromonas hydrophila. Moreover, Phβ-GBP-AgNPs exhibited less toxicity towards Artemia salina than chem-AgNPs during 24 h exposure period. As a result, Phβ-GBP-AgNPs may act as an alternative to commercial antibiotics and be considered an effective immunostimulant in treating skin lesions in intensive farming., (Copyright © 2021 Elsevier Ltd. All rights reserved.)