BackgroundUsing anaesthetics is an important application in aquaculture especially where the fish transportation, vaccination, grading, sorting activities and many other handling operations have been conducted during the different stages of production periods in the farms or hatcheries.ObjectivesThis study aimed to evaluate the efficacy of oregano essential oil (OO) as an anaesthetic for Nile tilapia and to determine the optimal concentration and post‐application stress effects compared to clove oil (CO).MethodsNile tilapia juveniles were exposed to different concentrations of OO (20–40–60–80–100 mg L−1) and CO (50 mg L−1) for different time periods to determine the optimal concentration and exposure time. After the effective concentration of OO was determined, in the second experiment, stress parameters (glucose, plasma cortisol) were analysed after 0, 2, 6, 12 and 24 h of application. The results compared to the control group and CO results.ResultsThe study found that OO which has carvacrol compound higher than 78% was an efficient anaesthetic for Nile tilapia, and the effective concentration was found to be 60 mg L−1. No significant differences were found between opercular beats in any concentration of OO and CO. Basal glucose level in blood without anaesthesia application and recorded at 39.33 mg dL−1 and significantly lower than OO and CO at first two sampling points, 0 and 2 h (p < 0.05). According to plasma cortisol level results, although CO experimental group showed secondary stress response at 12 h (17.91 ± 4.21 ng mL−1), OO and CO group cortisol levels decreased at 24 h after anaesthesia application 7.13 ± 0.14and 7.01 ± 0.54 ng mL−1, respectively, below the control group cortisol concentration (12.28 ± 1.81 ng mL−1).ConclusionsThese findings have important implications for the aquaculture industry as the use of OO as an anaesthetic could reduce the stress and mortality associated with traditional anaesthetics. Further research is needed to evaluate the efficacy of OO as an anaesthetic agent for other fish species and to determine the optimal concentration and exposure time for different species.