Three antioxidant properties (corresponding to the enzymes superoxide dismutase (SOD), catalase, and glutathione peroxidase) were measured in the livers of Scophthalmus maximus under different salinities/temperatures (5, 10, 20, 30, and 40‰/17, 20, 23, 25, and 28 °C). Split-plot (SP) analysis, additive main effects, and multiplicative interaction (AMMI) and genotype × environment interaction (GGE) biplots were used to analyze genotype × salinity/temperature interactions for antioxidant properties. The results of the SP analysis show that the activity of the three antioxidant properties was significantly (p < 0.01) affected by salinity/temperature, antioxidant, and salinity/temperature × antioxidant interaction. The results of the AMMI analysis for salinity reveal that the effect of genotype, salinity, and genotype × salinity interaction on antioxidant properties reached a significant level (p < 0.001); 92.1065%, 2.6256%, and 4.4360% of the total sum of squares for antioxidant property activity were attributable to the effects of genotype, salinity, and genotype × salinity interaction, respectively. The results of GGE biplot analysis for salinity reveal differences in the activity ranking of the three antioxidant properties under five salinities; this difference expands with an decrease or increase in salinity from 30‰ (optimum salinity). A salinity of 5‰ had the strongest ability to identify the three antioxidant properties. The five experimental salinities were divided into one region, and SOD activity was the highest in this region. In a comprehensive analysis of stability and activity, SOD had the best activity and stability. The results of AMMI analysis for temperature reveal that genotype, temperature, and genotype × temperature interaction had significant effects on the antioxidant properties (p < 0.001); 82.4720%, 4.0666%, and 12.0968% of the total sum of squares for antioxidant property activity were attributable to the effects of genotype, temperature, and genotype × temperature interaction, respectively. The results of GGE biplot analysis for temperature reveal a large difference in the activity ranking of antioxidant properties between 17 °C and the other four temperatures, while only small differences in the activity rankings were detected among the other four temperatures. The difference in the activity ranking of antioxidant properties was greatest between the temperatures of 17 and 20 °C. A temperature of 17 °C showed the strongest ability to distinguish the three antioxidant properties. Additionally, the five test temperatures were grouped into one region, and comprehensive analysis of activity and stability showed that SOD had the best activity and stability.

Turbot (Scophthalmus maximus) is an economically vital cold-water fish frequently challenged by summer heat and low salinity. However, the temporal response of physiological circadian rhythms to such long-term stress remains underexplored. This study investigated antioxidant, immune, and endocrine rhythms in turbot acclimated to control (16 °C, 30 ppt), high-temperature (23 °C), and low-salinity (10 ppt) conditions for 30 days. Subsequently, time-series sampling was performed every 4 h for 72 consecutive hours. Under optimal conditions, hepatic superoxide dismutase (SOD), serum alanine aminotransferase (ALT), and melatonin exhibited robust 24 h rhythms. Long-term stress disrupted this homeostasis through divergent mechanisms. Low-salinity stress induced “rhythmic remodeling,” maintaining balance via phase shifts or novel infradian (48–72 h) oscillations in thyroid hormones (T3, T4) and ALT, without oxidative damage. Conversely, high-temperature stress triggered “rhythmic collapse,” characterized by a loss of daily rhythms in SOD and ALT, sustained inflammation indicated by elevated acid phosphatase (ACP), metabolic depression (suppressed T3), and malondialdehyde accumulation. These findings demonstrate that heat stress poses a more destructive threat to circadian integrity than hyposmotic stress. Consequently, the rhythmic dynamics of ACP, ALT, T3, and T4 are identified as critical indicators of stress status, serving as potential biomarkers for screening stress-tolerant strains for selective breeding.

Quinoa germ meal (QGM) is a protein-rich by-product with potential as an alternative protein source; however, its effects on growth performance and intestinal health in marine carnivorous fish remain unclear. Juvenile turbot (Scophthalmus maximus) were fed five isonitrogenous (45.6% crude protein) and isolipidic (9.8% crude lipid) diets for 8 weeks: a fishmeal-based control diet (C) and four experimental diets in which fishmeal was replaced with QGM at 10% (Q10), 20% (Q20), 30% (Q30), and 40% (Q40). Growth performance, muscle proximate composition, intestinal histomorphology, and intestinal transcriptomic profiles were analyzed. Growth performance parameters, including final body weight, weight gain rate, specific growth rate, daily feed intake, and condition factor, decreased significantly with increasing QGM inclusion levels compared with the control (p < 0.05). Feed conversion ratio increased significantly only when replacement exceeded 30% (p < 0.05), while survival rate was unaffected (p > 0.05). Muscle crude lipid content was significantly reduced in all QGM-fed groups (p < 0.05), whereas crude protein, moisture, and ash contents were unchanged. Intestinal mucosal fold height increased in the Q30 and Q40 groups, while submucosal width decreased in the Q40 group (p < 0.05). Transcriptomic analysis revealed a dose-dependent increase in differentially expressed genes, mainly enriched in ribosome-related pathways, linoleic acid metabolism, and protein digestion and absorption. High dietary inclusion of QGM (>30%) impaired growth performance in juvenile turbot, whereas low inclusion levels (≤20%) exerted minimal adverse effects. Quinoa germ meal represents a potential alternative protein source, yet its effective utilization requires further optimization to maintain growth performance.

This study evaluated the effects of different water salinities (0, 10, 20 [control], 30, and 40; Practical Salinity Scale, dimensionless) over 56 days on growth performance, muscle biochemical composition, and antioxidant responses in serum and liver of Planiliza haematocheilus. After one-week pre-acclimation at 20, juveniles were directly transferred to the experimental salinities; thus, all groups except the 20 control experienced acute salinity transfer followed by 56-day exposure. Therefore, acute transfer stress and subsequent salinity exposure were confounded and could not be separated in this design. Survival was highest at 20 and lowest at 0. Weight gain rate and specific growth rate peaked at 30 and were lowest at 0. Under this direct-transfer protocol, the optimal salinity range for growth and survival was approximately 20–30 for these ~93 g juveniles. Hepatosomatic index differed significantly among salinity groups. Muscle moisture content was negatively correlated with salinity. Crude protein content was highest in the salinity-40 group, whereas crude fat was highest at 20 and lowest at 0; ash content did not differ significantly among treatments. Superoxide dismutase (SOD) activity in serum and liver showed no clear trend with salinity, but most groups differed significantly from the control. Liver SOD activity was highest at 10 and lowest at 30. Serum catalase (CAT) activity did not differ significantly among salinity groups, whereas liver CAT activity tended to decrease and then increase with rising salinity. Salinity had limited effect on serum malondialdehyde (MDA) content, except for a peak at 40, whereas liver MDA content decreased with increasing salinity and was significantly higher in the low-salinity groups. These results indicate that, under the direct-transfer protocol, P. haematocheilus juveniles (initial weight ~93 g) pre-acclimated to 20 and then directly transferred showed optimal growth and survival at 20–30, and that liver antioxidant parameters may serve as sensitive biomarkers of salinity stress. However, because acute transfer stress was confounded with subsequent 56-day salinity exposure, these findings are specific to this direct-transfer protocol and developmental stage and should not be extrapolated to gradual acclimation protocols or smaller juveniles.