In this study, the influence of temperature variation on the blood and oxidative stress parameters of pacu Piaractus mesopotamicus juveniles was evaluated. Fish were subjected to gradual changes of temperature, over 5 days, by 24°C until reaching to experimental temperatures 18, 21, 24 (control), 27 and 30°C. There were significant differences (p < .05) in glucose and lactate of blood in both the haematological relationships and oxidative stress parameters. Glucose levels increased at 30°C and decreased at 21°C. Lactate levels were highest at 18°C. While the haematocrit was higher at all altered temperatures, haemoglobin levels decreased at 21°C and increased at 30°C, causing alterations in haematimetric indices. In the liver, glutathione‐S‐transferase (GST) activity was reduced at 18°C while antioxidant capacity against peroxyl radicals (ACAP) was highest at 30°C. Thiobarbituric acid reactive substances (TBARS) increased at 18, 21 and 27°C. In the muscle, ACAP decreased at 27 and 30°C and TBARS increased at 30°C. The results indicate that the species shows sensitivity to temperature variations. Accordingly, we conclude that with short‐term temperature variations >3°C, it is possible to induce higher alterations in the observed parameters in pacu juveniles.