Avaliação do risco da exposição humana aos fungicidas triazóis no Sul de Minas Gerais: conectando biomonitoramento e toxicologia computacional

Abstract

Brazil stands as the world’s leading coffee producer, where the extensive use of pesticides is economically critical yet poses health and environmental risks due to their non-selective mechanisms of action. Specifically, triazole fungicides are widely used in agriculture to manage fungal diseases and are known to disrupt mammalian CYP450 and liver microsomal enzymes. This research establishes a framework for risk characterization of human exposure to triazole fungicides by internal-dose biomonitoring, biomarker measurements, and integration of high-throughput screening (HTS) data via computational toxicology workflows from the Integrated Chemical Environment (ICE). Volunteers from the southern region of Minas Gerais, Brazil, were divided into two groups: farmworkers and spouses occupationally and environmentally exposed to pesticides from rural areas (n = 140) and urban area individuals not occupationally exposed to serve as a comparison group (n = 50). Three triazole fungicides, cyproconazole, epoxiconazole, and triadimenol, were detected in the urine samples of both men and women in the rural group by gas chromatography coupled to mass spectrometry. Biomarker analysis indicated significant alterations in the rural group (Kruskal-Wallis p < 0.0001). Increased frequency of cellular changes associated with genotoxic effects by the oral mucosa cell cytome assay and oxidative imbalance, particularly among men occupationally exposed to pesticides. Additionally, elevated plasma bile acids indicating early and possibly reversible changes, as no significant differences were observed in liver enzyme levels (AST, ALT, and γ-GT). The hormones androstenedione and testosterone were significantly reduced in the farmworker group (Mann-Whitney p < 0.0001). The data show a significant inverse association of testosterone with cholesterol, LDL, VLDL, triglycerides, and glucose and a direct association with HDL (Spearman’s correlation p < 0.05). In the ICE workflow, active in vitro HTS assays were identified for the three measured triazoles and three other active ingredients from the pesticide formulations. The curated HTS data confirm bioactivities predominantly related to steroid hormone metabolism, cellular stress processes, and CYP450 enzymes impacted by fungicide exposure at occupationally and environmentally relevant concentrations based on the in vitro to in vivo extrapolation (IVIVE) models. To assess the risk associated with urinary triazole levels, Estimated Daily Intake (EDI) and Hazard Quotient (HQ) were calculated. The highest EDIs were observed for epoxiconazole, ranging from 0.47 to 6.31 μg/kg-bw/day for men and 0.49 to 8.77 μg/kg-bw/day for women in the exposed group. In the worst-case scenario, considering the highest detected urinary triazole value, the calculated HQ for epoxiconazole was 2.1 for men and 2.9 for women. These results characterize the potentially significant human health risk, particularly from the high frequency and intensity of exposure to epoxiconazole. This study showcases the critical role of biomonitoring and utility of computational tools in evaluating pesticide exposure and minimizing the adverse health effects of chronic exposure.