Assessment of genotoxic effects of organophosphate and carbamate pesticides by comet assay

Year 2022, , 136 - 142, 30.08.2022

Seher Karslı Çeppioğlu , Selma Yazar , Esra Fazilet İncedere Düzdağ , Türkan Yurdun

https://doi.org/10.26650/IstanbulJPharm.2022.1057224

Abstract

Background and Aims: Pesticide poisoning is the most widespread occupational hazard for agricultural workers in the de- veloping world, due to the extensive presence of pesticides in the environment. The aim of this study was to investigate the cytotoxicity and DNA damaging effects of organophosphosphate and carbamate pesticides.
Methods: In the present study, the cytotoxicity of chlorpyrifos methyl, azinphos ethyl, [(O-Ethyl O-(p-nitrophenyl) pheri- ylphosphonothioate] (EPN), aldicarb sulfone, and ethiofencarb were assessed by the trypan blue dye exclusion method. An alkaline comet assay was performed to assess the genotoxic effects of applied pesticides in human peripheral blood lym- phocytes.
Results: We demonstrated the cytotoxic effect of EPN following 30 and 120 min exposure at 100 µg/mL concentration. Al- though chlorpyrifos-methyl and azinphos ethyl seem to be safer concerning cytotoxicity compared to other pesticides, sig- nificantly higher DNA damage levels were determined after exposure of these pesticides for 120 min at 100 µg/mL concen- tration by in vitro comet assay. The potential DNA-damaging effects of these pesticides were sorted from high to low, as chlorpyrifos-methyl, aldicarb sulfone, EPN, and azinphos ethyl after 30 min of exposure, and were sorted as chlorpyrifos- methyl, azinphos ethyl, aldicarb sulfone, and EPN after 120 min of exposure. Our results revealed that these pesticides tend to increase DNA damage in a dose- and time-dependent manner.
Conclusion: The genotoxic effects of these widely used pesticides may cause prominent and serious health risks for hu- man populations; hence, the DNA-damaging potential of pesticides can lead to genotoxic risk and adverse health effects like cancer.

Keywords

Carbamate pesticides, Organophosphate pesticides, In vitro comet assay, Genotoxicity

Supporting Institution

Marmara University

Project Number

SAG-061/131102

References

• Abuwarda, H. N., Alashi, S. F., & Sharif, F. A. (2021). Investigating aneuploidy-inducing effect of Nemacur, Rogor, and Dursban in human peripheral blood lymphocyte cultures. Environmental and Molecular Mutagenesis, 62(5), 328-334. doi:10.1002/em.22445
• Ahmed, F. E. (2001). Analyses of pesticides and their metabolites in foods and drinks. TrAC Trends in Analytical Chemistry, 20(11), 649-661.
• Al-Samarraie, M. S. J., Karinen, R., Rognum, T., Hasvold, I., Stokke, M. O., & Christophersen, A. S. (2009). Lethal Poisoning with Ethiofen- carb and Ethanol. Journal of Analytical Toxicology, 33(7), 389-392. doi: 10.1093/jat/33.7.389
• Canna-Michaelidou, S., & Nicolaou, A. S. (1996). Evaluation of the genotoxicity potential (by Mutatox test) of ten pesticides found as water pollutants in Cyprus. Science of the Total Environment, 193(1), 27-35. doi:10.1016/s0048-9697(96)05322-3
• Cid, M. G., & Matos, E. (1984). Induction of sister-chromatid ex- changes in cultured human lymphocytes by aldicarb, a carba- mate pesticide. Mutation Research/Genetic Toxicology, 138(2), 175-179.
• Collins, A. R. (2014). Measuring oxidative damage to DNA and its repair with the comet assay. Biochimica et Biophysica Acta, 1840(2), 794-800. doi:10.1016/j.bbagen.2013.04.022
• Das, P., Shaik, A. P., & Jamil, K. (2007). Genotoxicity induced by pes- ticide mixtures: in-vitro studies on human peripheral blood lym- phocytes. Toxicology and Industrial Health, 23(8), 449-458.
• Ecobichon, D. J. (2001). Pesticide use in developing countries. Toxicology, 160(1-3), 27-33. doi: 10.1016/S0300-483x(00)00452-2
• Gaikwad, A. S., Karunamoorthy, P., Kondhalkar, S. J., Ambikapathy, M., & Beerappa, R. (2015). Assessment of hematological, bio- chemical effects and genotoxicity among pesticide sprayers in grape garden. Journal of Occupational Medicine and Toxicology, 10(1), 11.
• Hodgson, E., & Levi, P. E. (1996). Pesticides: an important but un- derused model for the environmental health sciences. Environ- mental Health Perspectives, 104(Suppl 1), 97.
• Jamil, K., Shaik, A. P., Mahboob, M., & Krishna, D. (2004). Effect of organophosphorus and organochlorine pesticides (monochro- tophos, chlorpyriphos, dimethoate, and endosulfan) on human lymphocytes in-vitro. Drug& Chemical Toxicology, 27(2), 133-144. doi:10.1081/dct-120030725
• Kaur, P., Radotra, B., Minz, R. W., & Gill, K. (2007). Impaired mito- chondrial energy metabolism and neuronal apoptotic cell death after chronic dichlorvos (OP) exposure in rat brain. Neurotoxicol- ogy, 28(6), 1208-1219.
• Kisby, G. E., Muniz, J. F., Scherer, J., Lasarev, M. R., Koshy, M., Kow, Y. W., & McCauley, L. (2009). Oxidative stress and DNA damage in agricultural workers. Journal of Agromedicine, 14(2), 206-214.
• McCauley, L. A., Lasarev, M. R., Higgins, G., Rothlein, J., Muniz, J., Ebbert, C., & Phillips, J. (2001). Work characteristics and pesticide exposures among migrant agricultural families: a community- based research approach. Environmental Health Perspectives, 109(5), 533.
• McKelvey-Martin, V. J., Green, M. H., Schmezer, P., Pool-Zobel, B. L., De Meo, M. P., & Collins, A. (1993). The single cell gel electropho- resis assay (comet assay): a European review. Mutation Research, 288(1), 47-63. doi:10.1016/0027-5107(93)90207-v
• Mehta, A., Verma, R. S., & Srivastava, N. (2008). Chlorpyrifos‐in- duced DNA damage in rat liver and brain. Environmental and Mo- lecular Mutagenesis, 49(6), 426-433.
• Mohanty, G., Mohanty, J., Jena, S. D., & Dutta, S. (2011). Geno- toxicity Testing in Pesticide Safety Evaluation. In M. Stoytcheva (Ed.) Pesticides in the Modern World-Pests Control and Pesticides Exposure and Toxicity Assessment (pp. 403-426). Rijeka, Croatia: InTech.
• Muniz, J. F., McCauley, L., Scherer, J., Lasarev, M., Koshy, M., Kow, Y., . . . Kisby, G. (2008). Biomarkers of oxidative stress and DNA dam- age in agricultural workers: a pilot study. Toxicology and Applied Pharmacology, 227(1), 97-107.
• Munoz-Quezada, M. T., Lucero, B. A., Iglesias, V. P., Munoz, M. P., Cornejo, C. A., Achu, E., . . . Villalobos, M. (2016). Chronic exposure to organophosphate (OP) pesticides and neuropsychological functioning in farm workers: a review. International Journal of Oc- cupational and Environmental Health, 22(1), 68-79. doi:10.1080/10 773525.2015.1123848
• Ni, Z., Li, S., Liu, Y., Tang, Y., & Pang, D. (1993). Induction of micro- nucleus by organophosphorus pesticides both in vivo and in vi- tro. Hua xi yi ke da xue xue bao= Journal of West China University of Medical Sciences= Huaxi yike daxue xuebao/[bian ji zhe, Hua xi yi ke da xue xue bao bian wei hui], 24(1), 82-86.
• Ojha, A., & Srivastava, N. (2014). In vitro studies on organophos- phate pesticides induced oxidative DNA damage in rat lympho- cytes. Mutation Research/Genetic Toxicology and Environmental Mutagenesis, 761, 10-17.
• Okonko, L., Ikpeme, E., & Udensi, O. (2016). Genotoxic Effect of Chlorpyrifos and Cypermethrin in Albino Rats. Research Journal of Mutagenesis, 6(1), 31.
• Petroianu, G. A., Nurulain, S. M., Hasan, M. Y., Kuca, K., & Lorke, D. E. (2015). Reversible cholinesterase inhibitors as pre-treatment for exposure to organophosphates: assessment using azinphos- methyl. Journal of Applied Toxicology, 35(5), 493-499. doi:10.1002/ jat.3052
• Proença, P., Teixeira, H., De Mendonça, M., Castanheira, F., Marques, E. P., Corte-Real, F., & Vieira, D. N. (2004). Aldicarb poisoning: one case report. Forensic Science International, 146, S79-S81.
• Rahman, M., Mahboob, M., Danadevi, K., Banu, B. S., & Grover, P. (2002). Assessment of genotoxic effects of chloropyriphos and acephate by the comet assay in mice leucocytes. Mutation Re- search/Genetic Toxicology and Environmental Mutagenesis, 516(1), 139-147.
• Sandal, S., & Yilmaz, B. (2011). Genotoxic effects of chlorpyrifos, cypermethrin, endosulfan and 2, 4‐D on human peripheral lym- phocytes cultured from smokers and nonsmokers. Environmental Toxicology, 26(5), 433-442.
• Shadnia, S., Azizi, E., Hosseini, R., Khoei, S., Fouladdel, S., Pajou- mand, A., . . . Abdollahi, M. (2005). Evaluation of oxidative stress and genotoxicity in organophosphorus insecticide formulators. Human & Experimental Toxicology, 24(9), 439-445.
• Singh, N. P., McCoy, M. T., Tice, R. R., & Schneider, E. L. (1988). A simple technique for quantitation of low levels of DNA damage in individual cells. Experimental Cell Research, 175(1), 184-191. doi:10.1016/0014-4827(88)90265-0
• Smith, G. J. (1987). Pesticide Use and Toxicology in Relation to Wildlife: Organophosphorus and Carbamate, Compounds. All U.S. Government Documents (Utah Regional Depository).
• Sun, X., Jin, Y., Wu, B., Wang, W., Pang, X., & Wang, J. (2010). Study on genotoxicity of aldicarb and methomyl. Huan jing ke xue= Huanjing kexue, 31(12), 2973-2980.
• Suratman, S., Edwards, J. W., & Babina, K. (2015). Organophos- phate pesticides exposure among farmworkers: pathways and risk of adverse health effects. Reviews on Environmental Health, 30(1), 65-79. doi:10.1515/reveh-2014-0072
• Ündeğer, Ü., & Başaran, N. (2002). Assessment of DNA damage in workers occupationally exposed to pesticide mixtures by the alkaline comet assay. Archives of Toxicology, 76(7), 430-436.
• van der Werf, H. M. (1996). Assessing the impact of pesticides on the environment. Agriculture, Ecosystems & Environment, 60(2-3), 81-96.
• Venkat, J. A., Shami, S., Davis, K., Nayak, M., Plimmer, J. R., Pfeil, R., & Nair, P. P. (1995). Relative genotoxic activities of pesticides evaluated by a modified SOS microplate assay. Environmental and Molecular Mutagenesis, 25(1), 67-76. doi:10.1002/em.2850250110
• Vigreux, C., Poul, J., Deslandes, E., Lebailly, P., Godard, T., Sichel, F., . . . Gauduchon, P. (1998). DNA damaging effects of pesticides measured by the single cell gel electrophoresis assay (comet as- say) and the chromosomal aberration test, in CHOK1 cells. Muta- tion Research/Genetic Toxicology and Environmental Mutagenesis, 419(1), 79-90.