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

Year 2021, Volume: 15 Issue: 1, 1 - 10, 29.04.2021

Emine Müge Karakayalı Duygu Kekeç Tuna Önal Mehmet İbrahim Tuğlu

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.

Keywords

Environmental Toxicity, MTT, Apoptosis, agricultural pesticide, neurotoxicity screening test

Thanks

We would like to thank to Izmir Democracy University.

References

• Yoshida M, McGregor D. Cyantraniliprole. In Pesticide residues in food 2013. Joint FAO/WHO Meeting on Pesticide Residues. Report of the Joint Meeting of the FAO Panel of Experts on Pesticide Residues in Food and the Environment and the WHO Core Assessment Group on Pesticide Residues. Geneva, Switzerland, September 17–26 2013. Rome: FAO; 2014. p. 131–76.
• EFSA Panel on Nutrition, Novel Foods and Food Allergens (NDA); Truck D, Castenmiller J, De Henauw S, Hirsch-Ernst KI, Kearney J, Maciuk A, Mangelsdorf I, McArdle HJ, Naska A, Pelaez C, Pentieva K, Siani A, Thies F, Tsabouri S, Vinceti M, Cubadda F, Frenzel T, Heinonen M, Marchelli R, et al. Scientific Opinion on the safety of dried yellow mealworm (Tenebrio molitor larva) as a novel food pursuant to regulation (EU) 2015/2283. EFSA NDA Panel (EFSA Panel on Nutrition, Novel Foods and Food Allergens). EFSA J 2021;19(1): e06343.1–29.
• European Food Safety Authority (EFSA). Conclusion on the peer review of the pesticide risk assesment of the active substance spiromesifen. EFSA J 2007;5(7):e105.1–69.
• European Food Safety Authority (EFSA). Conclusion on the peer review of the pesticide risk assessment of the active substance cyantraniliprole. EFSA J 2014;12899.e03814.1–249.
• Regueiro J, Olguín N, Simal-Gándara J, Suñol C. Toxicity evaluation of new agricultural fungicides in primary cultured cortical neurons. Environ Res 2015;140:37–44.
• El Merhie A, Salerno M, Toccafondi C, Dante S. Neuronal-like response of N2a living cells to nanoporous patterns of thin supported anodic alumina. Colloids Surf B Biointerfaces 2019;178:32–7.
• Wang H, Meng Z, Liu F, Zhou L, Su M, Meng Y, Zhang S, Liao X, Cao Z, Lu H. Characterization of boscalid-induced oxidative stress and neurodevelopmental toxicity in zebrafish embryos. Chemosphere 2020;238:124753.
• Çayır A, Coskun M, Coskun M. Micronuclei, nucleoplasmic bridges, and nuclear buds induced in human lymphocytes by the fungicide signum and its active ingredients (boscalid and pyraclostrobin). Environ Toxicol 2014;29:723–32.
• Bielza P, Moreno I, Belando A, Grávalos C, Izquierdo J, Nauen R. Spiromesifen and spirotetramat resistance in field populations of Bemisia tabaci Gennadius in Spain. Pest Manag Sci 2019;75:45–52.
• McLean WG. Holme AD, Janneh O, Southgate A, Howard CV, Reed MG. The effect of benomyl on neurite outgrowth in mouse N2A and human SH-SY5Y neuroblastoma cells in vitro. Neurotoxicology 1998;19:629–32.
• Smith SL, Sadler CJ, Dodd CC, Edwards G, Ward SA, Park BK, McLean WG. The role of glutathione in the neurotoxicity of artemisinin derivatives in vitro. Biochem Pharmacol 2001;15:409– 16.
• Vural K, Tuglu MI. Neurotoxic effect of statins on mouse neuroblastoma NB2a cell line. Eur Rev Med Pharmacol Sci 2011;15:985– 91.
• Ferrari E, Cardinale A, Picconi B, Gardoni F. From cell lines to pluripotent stem cells for modelling Parkinson’s disease. J Neurosci Methods 2020;340:108741.
• Axelrad JC, Howard CV, McLean WG. Interactions between pesticides and components of pesticide formulations in an in vitro neurotoxicity test. Toxicology 2002;173:259–68.
• Voorhees JR, Rohlman DS, Lein PJ, Pieper AA. Neurotoxicity in preclinical models of occupational exposure to organophosphorus compounds. Front Neurosci 2017;10:590.
• Vural K, Seyrek O. The neuroprotective effect of pioglitazone on NB2a mouse neuroblastoma cell culture. Kafkas Üniversitesi Veteriner Fakültesi Dergisi 2019;25:1–8.
• Klebe RJ, Chen T, Ruddle FH. Mapping of a human genetic regulator element by somatic cell genetic analysis. Proc Natl Acad Sci USA 1970;66:1220–7.
• LePage KT, Dickey RW, Gerwick WH, Jester EL, Murray TF. On the use of neuro-2a neuroblastoma cells versus intact neurons in primary culture for neurotoxicity studies. Crit Rev Neurobiol 2005;17:27–50.
• Lee MK, Nikodem VM. Differential role of ERK in cAMP-induced Nurr1 expression in N2A and C6 cells. Neuroreport 2004;15:99– 102.
• Axelrad JC, Howard CV, McLean WG. The effects of acute pesticide exposure on neuroblastoma cells chronically exposed to diazinon. Toxicology 2003;185:67–78.
• Mete M, Aydemir I, Ünlü Ünsal Ü, Duransoy YK, Tuğlu İM, Selçuki M. Neuroprotective effects of bone marrow-derived mesenchymal stem cellsand conditioned medium in mechanically injured neuroblastoma cells. Turk J Med Sci 2016;46:1900–7.
• Sachana M, Flaskos J, Alexaki E, Glynn P, Hargreaves AJ. The toxicity of chlorpyrifos towards differentiating mouse N2a neuroblastoma cells. Toxicol In Vitro 2001;15:369–72.
• Bjørling-Poulsen M, Andersen HR, Grandjean P. Potential developmental neurotoxicity of pesticides used in Europe. Environ Health 2008;7:50.
• Hargreaves AJ, Sachana M, Flaskos J. The use of differentiating N2a and C6 cell lines for studies of organophosphate toxicity. In: Aschner M, Suñol C, Bal-Price A, editors. Neuromethods, Vol 56: Cell culture techniques. New York ((NY): Springer; 2011. p. 269– 91.
• Bilir EK, Tutun H, Sevin S, Kismali G, Yarsan E Cytotoxic effects of Rhododendron ponticum L. extract on prostate carcinoma and adenocarcinoma cell line (DU145, PC3). Kafkas Üniversitesi Veteriner Fakültesi Dergisi 2018;24:451–7.
• Lau K, McLean WG, Williams DP, Howard CV. Synergistic interactions between commonly used food additives in a developmental neurotoxicity test. Toxicol Sci 2006;90:178–87.
• Qi Y, Cao X, Abd El-Aty AM, Ma C, Li H, Jiang Z, She Y, Wang S, Wang J, Yang S. A magnetic zeolitic imidazolate framework nanohybrid for fast and efficient extraction of clothianidin, imidacloprid, acetamiprid, and thiacloprid in water. J Nanosci Nanotechnol 2019; 19:3310–8.
• Damalas CA, Koutroubas SD. Farmers' exposure to pesticides: toxicity types and ways of prevention. Toxics 2016;4:1.
• Hinojosa MG, Gutiérrez-Praena D, Prieto AI, Guzmán-Guillén R, Jos A, Cameán AM. Neurotoxicity induced by microcystins and cylindrospermopsin: a review. Sci Total Environ 2019;10:668:547– 65.