Responses of hepatic biotransformation and antioxidant enzymes in Japanese medaka (Oryzias latipes) exposed to humic acid.

Humic substances, a major component of natural organic matter in surface waters, can induce biotransformation enzyme activities and influence antioxidant defense in fish. The study aimed to provide a molecular basis for the stress responses, the synthesis of biotransformation, and antioxidant enzymes in particular. Adult medaka fish (Hd-rR strain) were exposed to environmentally relevant concentrations of humic acid for 96 h. The actual humic acid concentrations in water were determined photometrically and expressed as organic carbon concentrations. Liquid chromatography with tandem mass spectrometry was used for protein profile analysis of medaka liver samples. The relative amount of isozymes was determined using the label-free quantification approach. Hepatic biotransformation enzyme activities were measured as well. Thus, ethoxyresorufin-O-deethylase activity showed a pronounced induction at the highest tested concentration (9.4 mg C/L). Various biotransformation and antioxidant isozymes responded to humic acid differently, reflecting a balanced interplay of proteins that ensures the metabolism of humic acid in fish liver. Some isozymes were not affected by humic acid. The study provides new insight into the molecular mechanisms of the fish stress response to the humic acid-related challenge.