Study of carbonate alkalinity-induced hepatic tissue damage in Hefang crucian carp (Carassius auratus) based on transcriptomic analysis

Abstract

This study investigated the effects of different sodium bicarbonate (NaHCO₃) concentrations (0 g/L, 1 g/L, and 3 g/L) on Hefang crucian carp (12.0 ± 1.1 g) over a 96-hour period. The experiment is divided into three groups, each with three replicates, and each replicate contains 30 fish. We employed a comprehensive approach integrating histology, physiological and biochemical assays, transcriptomics, as well as artificial intelligence (AI)-assisted analysis. This multifaceted method allowed us to examine changes in gill and liver morphology, osmoregulation, antioxidant capacity, immune response, and physiological metabolism. Results showed that gill and liver tissue damage increased with rising water alkalinity. Serum sodium (Na⁺), potassium (K⁺), blood ammonia, and gill Na⁺/K⁺-ATPase (NKA) levels increased significantly (p < 0.05). Hepatic antioxidant enzymes initially increased, then decreased with prolonged stress. Serum and liver immunoenzyme indices were higher in bicarbonate-treated groups compared to controls. Carbonate treatment altered lipid and glucose metabolism in both serum and liver. Transcriptome analysis, enhanced by large language models (LLMs), revealed differentially expressed genes (DEGs) significantly associated with ion binding, transport, apoptosis, and metabolism. In conclusion, excessive carbonate intake in fish alters serum physiological functions and affects hepatic metabolic functions. Crucian carp primarily regulate hepatic antioxidant systems, utilize carbohydrate breakdown for energy requirements, and employ lipids in osmoregulation. This study provides insights into fish adaptation to saline-alkaline environments and offers support for the development of aquaculture in saline-alkaline waters.