Investigation of methoxyfenozide insecticide genotoxicity with Allium and SMART methods
Abstract
Aim of study: To assess the genotoxicity of methoxyfenozide by the Allium test and Somatic Mutation and Recombination Test (SMART).
Area of study: The Institute of Science Laboratories at Nevsehir Haci Bektas Veli University.
Material and Methods: In the Allium test, onion root tips were exposed to methoxyfenozide concentrations of 0.005 mL/L, 0.01 mL/L, and 0.02 mL/L for 24 hours. Post exposure, root tips preparations were analyzed for Mitotic Index (MI) and Chromosome Aberrations (CA). In the SMART, 72 ± 4-hour old transheterozygous larvae, derived from the standard cross of multiple wing hairs (mwh) males and flare (flr3) virgin females, were exposed to methoxyfenozide at concentrations of 0.1 mL/L, 0.2 mL/L, and 0.4 mL/L in an instant medium. Preparations were made from the wings of emerging adult Drosophila to evaluate clone induction frequencies and wing spots.
Main results: The Allium test indicated a concentration-dependent decrease in MI values 5.88 ± 0.40, 4.71 ± 0.44, and 3.97 ± 0.47 at 0.005 mL/L, 0.01 mL/L, and 0.02 mL/L, respectively, compared to the negative control (7.28 ± 0.32). Methoxyfenozide induced common abnormalities such as irregularity, bridges, vagrant, stickiness, and disorientation. The SMART method showed higher clone induction frequencies in both mwh/flr3 and mwh/TM3 flies compared to the negative control. Single and twin clones were observed on the wings of transheterozygous flies.
Research highlights: The findings indicate that methoxyfenozide may be genotoxic to both plant and animal systems. However, further research is needed to fully understand the environmental impacts and potential risks to living organisms linked with the agricultural use of methoxyfenozide.
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Akyil D, Özkara A, 2015. Bitkilerde genotoksisite çalışmalarında yaygın olarak kullanılan parametreler ve uygulanan test sistemleri. Erzincan Üniv Fen Bilimleri Enstitüsü Dergisi 8(2):223–247. https://doi.org/10.18185/eufbed.83628
APVMA, 2002. Evaluation of the new active Methoxyfenozide in the product PRODIGY 240 SC INSECTICIDE The Australian Pesticides and Veterinary Medicines Authority (APVMA). https://www.apvma.gov.au/node/13851 [16 November 2023].
Barakat HM, Mahfoz HM, El-Atroush H, Mohammed MA, 2010. Comparative Study Between the Effects of the Synthetic Fungicide Mancozeb and the Biological Fungicide Plant Quard on Allium cepa plant. Egypt J Genet Cytol 39(1): 99–113.
Bıçakçı U, Çavuşoğlu K, Yapar K, Acar A, Yalçın E, 2017. Allium cepa L. Kök Ucu Hücrelerinde Diazinon Toksisitesinin Araştırılması. J Inst Sci Technol 7(3): 49–56. https://doi.org/10.21597/jist.2017.16150
Bonciu E, Firbas P, Fontanetti CS, Wusheng J, Karaismailoğlu MC, Liu D, Menicucci F, Pesnya DS, Popescu A, Romanovsky AV, Schiff S, Ślusarczyk J, de Souza CP, Srivastava A, Sutan A, Papini A, 2018. An evaluation for the standardization of the Allium cepa test as cytotoxicity and genotoxicity assay. Caryologia 71(3):191–209. https://doi.org/10.1080/00087114.2018.1503496
Budak A, 2005. The Detection of Genotoxic Activity of the Deltamethrin and Permethrin by Somatic Mutation and Recombination Test With. Ahi Evran Üniversitesi Kırşehir Eğit Fak Derg 6(1): 87–93.
Carton B, Smagghe G, Tirry L, 2003. Toxicity of two ecdysone agonists, halofenozide and methoxyfenozide, against the multicoloured Asian lady beetle Harmonia axyridis (Col., Coccinellidae). Journal of Applied Entomology 127(4): 240–242. https://doi.org/10.1046/j.1439-0418.2003.00734.x2
Chauhan LKS, Saxena PN, Gupta SK, 1999a. Cytogenetic effects of cypermethrin and fenvalerate on the root meristem cells of Allium cepa. Environ Exp Bot 42(3): 181–189. https://doi.org/10.1016/S0098-8472(99)00033-7
Chauhan LKS, Saxena PN, Gupta SK, 1999b. Effects of deltamethrin on the ultrastructures of the root meristem cells of Allium cepa. Pestic Biochem Physiol 64(3): 135–147. https://doi.org/10.1006/pest.1999.2416
Chen Y, Liu X, Dong F, Xu J, Wu X, Zheng Y, 2021. Characterization of the fate and distribution of methoxyfenozide in a water-plant-fish-sediment microcosm using a multimedia fugacity model. Sci Total Environ 755:142482. https://doi.org/10.1016/j.scitotenv.2020.142482
de Morais CR, Bonetti AM, Carvalho SM, de Rezende AAA, Araujo GR, Spanó MA, 2016. Assessment of the mutagenic, recombinogenic and carcinogenic potential of fipronil insecticide in somatic cells of Drosophila melanogaster. Chemosphere 165: 342–351. https://doi.org/10.1016/j.chemosphere.2016.09.023
de Morais CR, Carvalho SM, Carvalho Naves MP, Araujo G, de Rezende AAA, Bonetti AM, Spanó MA, 2017. Mutagenic, recombinogenic and carcinogenic potential of thiamethoxam insecticide and formulated product in somatic cells of Drosophila melanogaster. Mutat Res Genet Toxicol Environ Mutagen 823: 1–10. https://doi.org/10.1016/j.chemosphere.2017.08.108
Demir E, Kaya B, Kocaoglu Cenkci S, Cetin H, Marcos R, 2014. In vivo Genotoxicity of Four Synthetic Pyrethroids with Combinations of Piperonyl Butoxide (PBO) Using the Drosophila SMART Assay. Ekoloji Derg 23(92): 9–18. https://doi.org/10.5053/ekoloji.2014.922
European Food Safety Authority (EFSA), 2017. Peer review of the pesticide risk assessment of the active substance methoxyfenozide. EFSA Journal 15(9): e04978. https://doi.org/10.2903/j.efsa.2017.4722
El-Ghamery AA, El-Nahas AI, Mansour MM, 2000. The action of atrazine herbicide as an inhibitor of cell division on chromosomes and nucleic acids content in root meristems of Allium cepa and Vicia faba. Cytologia 65(3): 277–287. https://doi.org/10.1508/cytologia.65.277
The Food and Agriculture Organization (FAO), 2017. The future of food and agriculture and challenges. http://www.fao.org/3/a-i6583e.pdf
Feulgen R, Rossenbeck H, 1924. Mikroskopisch-chemischer Nachweis einer Nucleinsäure vom Typus der Thymonucleinsäure und die darauf beruhende elektive Färbung von Zellkernen in mikroskopischen Präparaten. Hoppe-Seyler’s Z Physiol Chem 135(5–6): 203–248.
Firbas P, Amon T, 2014. Chromosome damage studies in the onion plant Allium cepa L. Caryologia 67(1): 25–35. https://doi.org/10.1080/00087114.2014.891696
Fisher A, Colman C, Hoffmann C, Fritz B, Rangel J, 2018. The effects of the insect growth regulators methoxyfenozide and pyriproxyfen and the acaricide bifenazate on honeybee (Hymenoptera: Apidae) forager survival. J Econ Entomol 111(2): 510–516. https://doi.org/10.1093/jee/tox347
Fiskesjö G, 1985. The Allium test as a standard in environmental monitoring. Hereditas 102(1): 99–100. https://doi.org/10.1111/j.1601-5223.1985.tb00471.x
Frei H, Würgler FE, 1988. Statistical methods to decide whether mutagenicity test data from Drosophila assays indicate a positive, negative, or inconclusive result. Mutat Res Environ Mutagen and Relat Subj 203(4): 297–308. https://doi.org/10.1016/0165-1161(88)90019-2
Garcia FP, Cortés Ascencio SY, Gaytan Oyarzun JC, Ceruelo Hernandez A, Vazquez Alavarado P, 2012. Pesticides: classification, uses and toxicity. Measures of exposure and genotoxic risks. J Res Environ Sci Toxicol 1(11): 279–293.
Giraudo M, Califano J, Hilliou F, Tran T, Taquet N, Feyereisen R, Le Goff G, 2011. Effects of hormone agonists on Sf9 cells, proliferation and cell cycle arrest. PLoS ONE 6(10): e25708. https://doi.org/10.1371/journal.pone.0025708
Graf U, 1995. Analysis of the relationship between age of larvae at mutagen treatment and frequency and size of spots in the wing somatic mutation and recombination test in Drosophila melanogaster. Experientia 51(2): 168–173. https://doi.org/10.1016/0165-1161(92)90032-H
Graf U, van Schaik N, 1992. Improved high bioactivation cross for the wing somatic mutation and recombination test in Drosophila melanogaster. Mutat Res Environ Mutagen and Relat Subj 271(1): 59–67. https://doi.org/10.1016/0165-1161(92)90032-H
Graf U, Würgler FE, Katz AJ, Frei H, Juon H, Hall CB, Kale PG, 1984. Somatic mutation and recombination test in Drosophila melanogaster. Environ Mutagen 6(2): 153–188. https://doi.org/10.1002/em.2860060206
Grant WF, Salamone MF, 1994. Comparative mutagenicity of chemicals selected for test in the International Program on Chemical Safety’s collaborative study on plant systems for the detection of environmental mutagens. Mutat Res Fundam Mol Mech Mutagen 310(2): 187–209. https://doi.org/10.1016/0027-5107(94)90113-9
Hamaidia K, Soltani N, Hamaidia CK, 2016. Ovicidal activity of an insect growth disruptor (methoxyfenozide) against Culex pipiens L. and delayed effect on development. J Entomol Zool Stud 4(4): 1202–1207.
Haouari-Abderrahim J, Rehimi N, 2014. Biological and reproduction activities of mosquito larvae of Culiseta morsitans (Theobald) after treatment by ecdysone agonist methoxyfenozide. Annu Res Rev Biol 4(24): 4152–4165. https://doi.org/10.9734/ARRB/2014/11366
Inceer H, Hayirlioglu-Ayaz S, Ozcan M, 2009. Genotoxic effects of the insecticide cypermethrin on the root meristem cells of sunflowers (Helianthus annuus L.). Bull Environ Contam Toxicol 83(5): 652–656. https://doi.org/10.1007/s00128-009-9856-8
Kalefetoğlu Macar T, 2021. Investigation of cytotoxicity and genotoxicity of abamectin pesticide in Allium cepa L. Environ Sci Pollut Res 28(2): 2391–2399. https://doi.org/10.1007/s11356-020-10708-0
Kastenbaum MA, Bowman KO, 1970. Tables for determining the statistical significance of mutation frequencies. Mutat Res 9: 527–549. https://doi.org/10.1016/0027-5107(70)90038-2
Kwiatkowska M, Reszka E, Woźniak K, Jabłońska E, Michałowicz J, Bukowska B, 2017. DNA damage and methylation induced by glyphosate in human peripheral blood mononuclear cells (in vitro study). Food Chem Toxicol 105: 93–98. https://doi.org/10.1016/0027-5107(70)90038-2
Lindsley D, Zimm G, 1992. The genome of Drosophila melanogaster. Academic Press.
Liu Y, Wang X, Wang S, 2024. Environmental fate and safety analysis of methoxyfenozide application to control litchi and longan pests. Environ Sci Pollut Res 31(25): 37316–37325. https://doi.org/10.1007/s11356-024-33677-0
Marcos R, Carmona ER, 2019. The Wing-Spot and the Comet Tests as useful assays for detecting genotoxicity in Drosophila. Methods Mol Biol 2031: 337–348. https://doi.org/10.1007/978-1-4939-9646-9_19
McGill M, Pathak S, Hsu TC, 1974. Effects of ethidium bromide on mitosis and chromosomes: a possible material basis for chromosome stickiness. Chromosoma 47(2): 157–166. https://doi.org/10.1007/BF00331803
Mesi A, Kopliku D, 2013. Cytotoxic and Genotoxic Potency Screening of Two Pesticides on Allium cepa L. Procedia Technol 8: 19–26. https://doi.org/10.1016/j.protcy.2013.11.005
Mesi AD, Kopliku D, 2015. Cyto- and Genotoxic Activity of Pesticide Cypermex Plus 550 EC on Allium cepa L. J Int Environ Appl Sci 10(4): 475–481.
Mosallanejad H, Badisco L, Swevers L, Soin T, Knapen D, Broeck J Vanden, Smagghe G, 2010. Ecdysone signaling and transcript signature in Drosophila cells resistant against methoxyfenozide. J Insect Physiol 56(12): 1973–1985. https://doi.org/10.1016/j.jinsphys.2010.08.022
Osaba L, Aguirre A, Alonso A, Graf U, 1999. Genotoxicity testing of six insecticides in two crosses of the Drosophila wing spot test. Mutat Res Genet Toxicol Environ Mutagen 439(1): 49–61. https://doi.org/10.1016/S1383-5718(98)00173-9
Pitchakarn P, Inthachat W, Karinchai J, Temviriyanukul P, 2021. Human hazard assessment using Drosophila wing spot test as an alternative in vivo model for genotoxicity testing—A review. Int J Mol Sci 22(18): 9932. https://doi.org/10.3390/ijms22189932
Rank J, Nielsen MH, 1993. A modified Allium test as a tool in the screening of the genotoxicity of complex mixtures. Hereditas 118: 49–53. https://doi.org/10.1111/j.1601-5223.1993.t01-3-00049.x
Sapbamrer R, Hongsibsong S, Sittitoon N, Amput P, 2019. DNA damage and adverse neurological outcomes among garlic farmers exposed to organophosphate pesticides. Environ Toxicol Pharmacol 72: 103241. https://doi.org/10.1016/j.etap.2019.1032411
Sharma S, Vig A, 2012. Genotoxicity of Atrazine, Avenoxan, Diuron and Quizalofop-P-ethyl Herbicides using the Allium cepa Root Chromosomal Aberration Assay. Terr Aquat Environ Toxicol 6(2): 90–95.
Toğay VA, Aşci Çelik D, 2022. Thiacloprid’in insan lenfositlerinde DNA hasarına etkisi. Med J Suleyman Demirel Univ 29(4): 597–602. https://doi.org/10.17343/sdutfd.1200362
Tudi M, Ruan HD, Wang L, Lyu J, Sadler R, Connell D, Chu C, Phung DT, 2021. Agriculture development, pesticide application and its impact on the environment. Int J Environ Res Public Health 18(3): 1112. https://doi.org/10.3390/ijerph18031112
Varona-Uribe ME, Torres-Rey CH, Díaz-Criollo S, Palma-Parra RM, Narváez DM, Carmona SP, Briceño L, Idrovo AJ, 2016. Exposure to pesticide mixtures and DNA damage among rice field workers. Arch Environ Occup Health 71(1): 3–9. https://doi.org/10.1080/19338244.2014.910489
Wu SP, Sun LL, Zhong H, Yan SC, Wu HQ, Wang ZY, Cao CW, 2015. Effects of methoxyfenozide on ecdysone receptor complex expression in larval Asian gypsy moth. Open Life Sci 10(1): 269–277. https://doi.org/10.1515/biol-2015-0025
Yildiz M, Arıkan SE, 2008. Genotoxicity testing of quizalofop-P-ethyl herbicide using the Allium cepa anaphase-telophase chromosome aberration assay. Caryologia 61(1): 45–52. https://doi.org/10.1080/00087114.2008.10589608
Zarate N, Díaz O, Martínez AM, Figueroa JI, Schneider MI, Smagghe G, Viñuela E, Budia F, Pineda S, 2011. Lethal and sublethal effects of methoxyfenozide on the development, survival, and reproduction of the fall armyworm, Spodoptera frugiperda (J. E. Smith) (Lepidoptera: Noctuidae). Neotrop Entomol 40(1): 129–137. https://doi.org/10.1590/S1519-566X2011000100020
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