Effects of Difenoconazole on Tubifex tubifex: Antioxidant Activity, Insights from GUTS Predictions, and Multi-Biomarker Analysis

The increasing demand for agricultural products has led to a rise in pesticide use, resulting in the pollution of aquatic habitats and raising significant health concerns for both aquatic life and humans. Difenoconazole, a triazole fungicide, is becoming increasingly popular in agriculture, yet its effects on non-target organisms, such as annelids, are not well understood. This study aimed to investigate the toxicological effects of difenoconazole and assess its potential impact on toxicity biomarkers, using <i>Tubifex tubifex</i> as a model organism, to better understand the ecotoxicity of difenoconazole on freshwater annelids. The 96-h LC₅₀ value of difenoconazole was determined to be 2.68 mg/L. Sublethal concentrations (10% and 20% of the 96-h LC₅₀ value; 0.268 and 0.536 mg/L, respectively) caused significant changes in the activities of oxidative stress enzymes. A concentration- and time-dependent decrease in the activities of superoxide dismutase (SOD), catalase (CAT), and glutathione transferase (GST) was observed compared to control organisms. Additionally, malondialdehyde (MDA) concentrations increased throughout the exposure period. An Integrated Biomarker Response (IBR) assessment was used to characterize and illustrate the impact of difenoconazole on <i>T. tubifex</i>. In conclusion, exposure to this fungicide appears to reduce the survival rate of <i>T. tubifex</i> at acute levels and disrupt its normal behavioral patterns. Moreover, it alters oxidative stress enzyme levels during sublethal exposure. Long-term exposure to the fungicide could potentially have population-level consequences, including a reduction in the number of individuals within a population.