Mixture effects of imidacloprid and difenconazole on enzymatic activity and gene expression in small yellow croakers (Larimichthys polyactis)

Agrochemicals usually exist as mixtures in aqueous ecosystems and have harmful impacts on the natural environment. Nonetheless, the combined effects and underlying mechanisms of agrochemicals on aqueous organisms remain poorly understood. In the present study, the interactive effects of imidacloprid (IMI) and difenconazole (DIF) on the embryos of small yellow croakers (Larimichthys polyactis) were assessed using various toxicological assays, including acute toxicity, enzymatic activity, and gene expression changes. The results showed that DIF (72-h LC50 value of 0.20 mg L−1) had higher toxicity than IMI (72-h LC50 value of 12.5 mgL−1). Simultaneously, combinations of IMI and DIF exerted synergistic acute effects on the embryos of L. polyactis. In addition, the SOD, CAT, GST, and CarE activities were noticeably altered in most single and mixed exposures, relative to the untreated control. The expression of four genes (cyp19a1b, ngln2, klf2a, and socs3a) related to the immune system, endocrine system, and neurodevelopment was also surprisingly altered when the embryos of L. polyactis were subjected to individual and combined exposures relative to the untreated control. Changes in enzymatic activity and gene expression might provide early warning indices for the identification of agrochemical co-exposure. The results of this study provide valuable insights into the comprehensive toxicity of agrochemical mixtures to L. polyactis. Further studies on the long-term effects of agrochemical mixtures on marine fish should be conducted to formulate definitive conclusions concerning hazards.