The cockroach Gromphadorhina coquereliana can survive at low temperatures under extensive periods of cold stress. To assess energy management and insect adaptation in response to cold, we measured mitochondrial activity and oxidative stress in muscle and fat body tissues from G. coquereliana under a fluctuating thermal regime (FTR; stressed at 4°C for 3 h on 3 consecutive days, with or without 24 h recovery). Compared with our earlier work showing that a single exposure to cold significantly affects mitochondrial parameters, here, repeated exposure to cold triggered an acclimatory response, resulting in unchanged mitochondrial bioenergetics. Immediately after cold exposure, we observed an increase in the overall pool of ATP and a decrease in typical antioxidant enzyme activity. We also observed decreased activity of uncoupling protein 4 in muscle mitochondria. After 24 h of recovery, we observed an increase in expression of antioxidant enzymes in muscles and the fat body and a significant increase in the expression of UCP4 and HSP70 in the latter. This indicates that processes related to energy conversion and disturbance under cold stress may trigger different protective mechanisms in these tissues, and that these mechanisms must be activated to restore insect homeostasis. The mitochondrial parameters and enzymatic assays suggest that mitochondria are not affected during FTR but oxidative stress markers are decreased, and a 24 h recovery period allows for the restoration of redox and energy homeostasis, especially in the fat body. This confirms the crucial role of the fat body in intermediary metabolism and energy management in insects and in the response to repeated thermal stress.

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