Integrated Transcriptomic and Metabolomic Analyses Reveal Physiological and Hepatic Metabolic Responses of Largemouth Bass (Micropterus salmoides) to Subacute Saline–Alkaline Stress

Freshwater scarcity makes saline–alkaline water essential for sustainable aquaculture. Current research has primarily focused on individual salinity or alkalinity effects on fish, with limited studies addressing their interactive effects. We found significant synergistic toxicity between salinity and alkalinity (r ≈ −0.950/−0.925) in largemouth bass (Micropterus salmoides), demonstrating higher salinity levels corresponding to lower 96-h median lethal concentration (96 h LC50) values for alkalinity, and vice versa. A subsequent 56-day subacute stress trial (salinity: 6‰; alkalinity: 20 mmol/L) assessed the impact on largemouth bass through growth efficiency, histopathology, biochemical assays, transcriptomics, and metabolomics, comparing a saline–alkaline group (SA) with a normal control group (NC). There were no significant differences in growth or survival between the SA and NC groups, but the SA group exhibited pathological changes in gill and liver tissues. Biochemically, the SA group exhibited elevated malondialdehyde, glutathione, and blood urea nitrogen levels, whereas glutathione peroxidase activity significantly decreased. Integrated transcriptomics and metabolomics analyses demonstrated that saline–alkaline stress disrupts lipid, amino acid, and steroid metabolism in largemouth bass, affecting steroid biosynthesis, fatty acid metabolism, glycerophospholipid metabolism, and cysteine and methionine metabolism pathways. Fish adapt by adjusting gene expression and metabolite levels to maintain metabolic balance. This study highlights adaptive mechanisms and applications for sustainable largemouth bass culture in saline–alkaline environments.