Investigation of the moderate toxicity of agricultural pesticides cyantraniliprole, boscalid and spiromesifen in vitro using neurotoxicity screening test

Abstract

Objectives: Although industrial products used as agricultural pesticides are considered safe, they are likely to lead to chronic problems due to their long-term effects. The neurotoxicity screening test (NST) is a method based on the inhibition of neurite extension of neurons that do not not die with toxic effects. In this study, we aimed to investigate the moderate neurotoxic effects and reveal the potential dangers of agricultural pesticides in vitro using NST. Methods: Cyantraniliprole, boscalid and spiromesifen were used as agricultural pesticides on the mouse neuroblastoma cell line N2a. Neurite extension of neurons was performed by taking them into the proliferation medium followed by the differentiation medium. Cell viability and proliferation were analyzed using the MTT test. The percentage of neurite inhibition was calculated by measuring neurite outgrowth by NST. Oxidative stress was analyzed by NOS staining with h-score and apoptosis was shown using the apoptotic index in TUNEL staining. Results: Cyantraniliprole, boscalid and spiromesifen at high concentrations caused neurite inhibition, decreased proliferation and reduced the viability of cultured neurons. These agricultural pesticides were found to be significantly moderate toxic for neurons by increasing oxidative stress and apoptosis. Conclusion: We conclude neurite inhibition may be important in early recognition for detecting and preventing the neurotoxic effect of pesticides, and NST is an important in vitro test that can predict the long-term effects of neurotoxic agents. In the present study, we observed cyantraniliprole, boscalid and spiromesifen had moderate neurotoxic effects in varying degrees using NST. This means that pesticides may behave toxic even in permissible limits for chronic exposure.