This study was carried out to evaluate combined and singular effects of ethanolic extract of Persian shallot (Allium hirtifolium Boiss) and synbiotic Biomin® IMBO on growth performance, innate immune responses, and antioxidant defense in zebrafish (Danio rerio). Fish with initial weight of 151.90 ± 0.31 mg were allocated in 21 10-L glass aquariums. The experimental groups were as follows: T1, control (without any supplementation); T2, 1% synbiotic; T3, 3% synbiotic; T4, 1% Persian shallot (as a medical plant); T5, 3% Persian shallot; T6, 1% Persian shallot and 1% synbiotic; T7, 3% Persian shallot and 3% synbiotic. At the end of the experiment (60 days), all treatments significantly showed higher final weight (FW), weight gain (WG), WG (%), and specific growth rate (SGR) compared with the fish fed on control diet. Furthermore, both synbiotic Biomin® IMBO and Persian shallot significantly improved intestine immune parameters including lysozyme, alternative complement hemolytic activity (ACH50), total immunoglobulin (total Ig), and myeloperoxidase (MPO) of ze-brafish compared to fish fed on control diet (p < 0.05). Also, in all experimental groups, hepatic catalase (CAT), superoxide dismutase (SOD), glutathione peroxidase (GPx), and glutathione reduc-tase (GR) activities significantly increased compared to the control group. Whereas, the highest MDA level was observed in the control group compared to the treatments (p < 0.05). Moreover, skin mucus immune parameters of zebrafish have been noticeably improved with synbiotic Biomin® IMBO and Persian shallot compared to fish fed on the control diet (p < 0.05). The results indicate that synbiotic or Persian shallot supplemented diet could enhance the general health status of the zebrafish. © 2021 by the authors. Licensee MDPI, Basel, Switzerland.