© 2018 John Wiley & Sons Ltd This study explores the anaesthetic activity of Alpinia galanga oil (AGO) in fish. Cyprinus carpio (koi carp) was used as a fish model. It was found that the induction time to stage 3 anaesthesia and the recovery time of the fish after exposure to AGO were exponentially and polynomially correlated to AGO concentrations. The viability of normal blood cells of koi carp anaesthetized with 500 mg/L AGO was found to be higher than 90% for normal red blood cells and white blood cells and 89% for peripheral blood nuclear cells indicating nontoxicity of AGO to the fish. A concentration of 300 mg/L of AGO was the most suitable for anaesthetizing koi carp due to the safety and effectiveness aspects as being ideally fitted to anaesthetic criteria. This concentration gave the induction time of 205.55 ± 5.07 s and the recovery time of 202.50 ± 9.30 s. Determination of stress biomarker such as blood cortisol and glucose as well as gene expression showed that the blood cortisol level of the fish anaesthetized with AGO was similar to normal levels. Moreover, blood glucose level was significantly less increased than those anaesthetized with tricaine methanesulfonate. Gene expressions of the fish cortisol receptor, cytochrome oxidase subunit1, heat shock protein 70 and Na + /K + -ATPaseα 3 were significantly reduced after exposure to AGO indicating the advantages of AGO on fish stress reduction. Thus, AGO is a promising natural source for an alternative fish anaesthetics.