"Fishcide" Effect of the Fungicide Difenoconazole in Freshwater Fish (labeo Rohita): A Multi-Endpoint Approach.

Difenoconazole is widely used to protect crops, fruits, and vegetables. However, this fungicide can enter aquatic environments and cause harmful effects to non-target organisms and induce little-known biological disorders. Thus, aiming to expand our knowledge about the ecotoxicity of difenoconazole on freshwater ichthyofauna, we aimed to determine the median lethal concentration (LC50) of difenoconazole and evaluate its possible impacts from different toxicity biomarkers, using freshwater fish Labeo rohita as a model system. Using the probit analysis method, the 96 h LC50 value of difenoconazole in the fish was calculated as 4.5 mg L-1. Posteriorly, fish were exposed to two sublethal concentrations (0.45 mg L-1 1/10th and 0.9 mg L-1 1/5th LC50 value) for 21 days. A significant reduction of superoxide dismutase (SOD) and catalase (CAT) activity was noted in the gill, liver, and kidneys of fish compared to the control groups. The level of glutathione-S-transferase (GST) and lipid peroxidation (LPO) activity was higher in all vital tissues of difenoconazole-treated fish. Histological alterations in the gill include epithelial lifting, lamellar fusion, hypertrophy, and epithelial necrosis. At the same time, the liver showed pyknotic nucleus, vacuolation, cellular edema and tubular necrosis, shrinkage of glomeruli, vacuolation, and pyknotic nuclei in the kidney. DNA damage was increased significantly with tail formation based on the concentration and time-dependent manner. Therefore, our study confirms that the exposure of L. rohita to difenoconazole induces negative biological consequences and sheds light on the danger