Intensive recirculating systems are a fast-developing sector of aquaculture. While several warm-water fish have been reared in aquaponics, almost no data are available for cold-water species. The determination of nitrate toxicity thresholds in recirculating aquaculture is crucial. Different pollutants are typically more toxic at elevated temperatures. We investigated the performance of Oncorhynchus mykiss under two different nitrate levels and two temperatures. We applied a 2 × 2 factorial design, where fish (9.78 ± 0.51 g) were exposed to nitrate concentrations of 40 or 110 mg/L NO3− and to temperatures of 17 °C or 21 °C for 20 days. This study focused on understanding the physiological responses of rainbow trout to relatively low nitrate levels under heat stress in order to investigate the feasibility of integrating this species into commercial aquaponics. The growth, condition, and expression of genes involved in metabolism, heat shock, antioxidant, and immune response were assessed in the liver, together with the activities of enzymes related to glucose and fatty acid metabolism. High nitrate levels at 17 °C affected the condition but did not alter growth, leading to increased glycolytic potential and, occasionally, a greater reliance on lipid oxidation. Antioxidant defense was mainly induced due to high nitrates and the similar expression patterns of antioxidant genes observed under high nitrate at both 17 °C and 21 °C. Warm exposure decreased condition and growth, leading to greatly reduced glucokinase transcription, irrespective of the nitrate levels. Exposure to 21 °C and high nitrate led to equivalent growth and condition as well as to a milder inflammatory response combined with metabolic readjustments (enhancement of glycolytic and lipid oxidation pathways) compared to the low nitrates at 21 °C. Based on the results, rearing at a temperature close to 21 °C should be avoided for fingerling growth, while NO3− concentration until 110 mg/L may not have severe impacts on fingerling health and growth at 17 °C. In addition, rainbow trout fingerlings can tolerate a 20-day exposure at 21 °C and NO3− up to 110 mg/L. Additional factors should always be considered, such as specific water quality parameters, for a comprehensive approach to assessing the feasibility of rainbow trout aquaculture in aquaponics.
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