The herbicide market is expanding rapidly due to the global increase in herbicide usage. Dairy cows are susceptible to herbicide exposure through the ingestion of contaminated plants, which can adversely affect the mammary gland health and reduce milk production. Dinitramine, a synthetic herbicide in the dinitroaniline family, is typically used to control weeds by inhibiting their sprouting and root development. While previous studies have demonstrated the cytotoxicity of dinitramine in aquatic organisms, research on its toxicity in cattle is limited. In this study, bovine mammary epithelial cells (MAC-T) were used to verify the detrimental effects on dairy cows, especially on the mammary glands. First, we evaluated the cytotoxic effects of dinitramine on MAC-T cells and examined various cellular responses to dinitramine treatment, including alterations in apoptotic cells, mitochondrial dysfunction, and calcium dysregulation. Moreover, the expression levels of AKT and MAPK signaling proteins were confirmed in response to dinitramine treatment. Alterations in the mRNA levels of genes related to milk production and inflammatory response following dinitramine exposure were evaluated using quantitative PCR. Finally, we assessed the binding affinity between dinitramine and the target proteins using in silico molecular docking analysis. Overall, the cumulative evidence of the various toxic effects of dinitramine on MAC-T cells suggests its potential to reduce both milk yield and quality.
Keywords: Apoptosis; Bovine mammary gland; Calcium; Dinitramine; Mitochondrial dysfunction; Molecular docking.
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