The present study evaluated the anaesthetic activity of tobacco (Nicotiana tabacum) leaf dust in fish. GIFT tilapia was used as an experimental animal with the total study duration of 24 hours. The fish size of 4.58±0.68 g was randomly distributed into the 20-liter plastic tub at a stocking density of 10 fishes per container. Anaesthetics were added into experimental tubs of seven different groups viz. one control (0 mg/L), followed by six treatment groups (25, 50, 75, 100, 125 and 150 mg/L) in triplicates. Results showed that an increase in the concentration of tobacco leaf dust shortened the induction time, but delayed the recovery time in fish (p<0.05). There was no mortality observed in control (without sedative) and up to the dosage of 25 to 100 mg/L, but after that mortality ensued from 6 to 16% in the groups exposed to a high dose of more than 100 mg/L (p<0.05). The temperature in the tanks showed no difference among all groups, while DO, pH, and C02 were affected by anaesthetics given (p<0.05). The results of this study are encouraging that tobacco leaf dust has the potential to sedate the fish, and it minimizes the water quality concerns at a correct level. Based on the result of this study, tobacco leaf dust at the levels of 25 to 100 mg/L can be recommended to the fish farmers for safe and successful transportation of GIFT tilapia without causing the mortality.