Akut Difenokonazol’ün Daphnia magna üzerindeki Genotoksik Etkileri

Abstract

Difenokonazol, tarımda sıkça kullanılan bir fungisit olup; sucul canlılar için zararlı olabilmektedir. Difenokonazol’un sucul canlılarda olan toksisitesi henüz iyi bilinmemektedir. Triazol mantar ilaçları arasında, Difenokonazol (PEN) birçok ülkede en yaygın kullanılan fungusitlerden biridir. Bu çalışmada, Daphnia magna üzerinde farklı dozlarda Difenokonazolün genotoksik etkisi araştırıldı. Ayrıca Kontrol Grubu ve diğer doz grublarıyla yapılan çalışma, Ekonomik İşbirliği ve Kalkınma Örgütü (OECD) standartları 202 ve 212 (OECD, 2012, 2009) tarafından belirlenen standart metodolojiye uygun olarak gerçekleştirildi. Çalışmada, Daphnia magna 10 gün boyunca kontrol grubu (0 mg/L) ve üç farklı dozda Difenokonazole (1.00, 1.5, 2.5 mg/L) maruz bırakıldı. Deneyin sonunda, dokuların Hasar Frekansı (%), Arbitrary Birim (%), ve Genetik Hasar İndeksi (%) belirlemek için Comet testi kullanıldı. Doz gruplarından 2.5 mg L-1 grubu, Daphnia magna üzerinde en yüksek hasar frekansına sahip olduğu(45.33±1.52) (p<0.001) tespit edildi. Elde ettiğimiz bulguların Difenokonazol’un maruziyeti sonrasında Daphnia magna'da DNA hasar düzeylerinde önemli bir artış olduğunu göstermesine bağlı olarak kullanılan bu fungisitin daphnidler için genotoksik olabileceğini işaret etmiştir.

Keywords

• pestisit
• Difenkonazol
• komet test
• DNA Hasarı

Genotoxic Effects of Acute Difenoconazole Exposure on Daphnia magna

Abstract

Difenoconazole, a fungicide often used in agriculture, is harmful to aquatic organisms. However, its toxicity to aquatic organisms is not yet well recognized. Among the triazole fungicides, Difenoconazole (PEN) is one of the most extensively used in many countries. In this study, the genetic impacts of different Difenoconazole doses on Daphnia magna was investigated. Experiments involving the control group and treatments were executed in compliance with the standard methodology outlined by the Economic Cooperation and Development Organization (OECD) standards 202 and 212 (OECD, 2012, 2009). In the present study, Daphnia magna were exposed to a control group (0 mg/L) and three different doses of (1.00, 1.5, 2.5 mg/L) difenoconazole for a period of 10 days. At the end of the experiment, comet assay was used to determine the damage frequency (%), Arbitrary unit (%), and Genetic damage index (%) of tissues. The 2.5 mg L-1 group showed notably greater damage frequencies (45.33±1.52) on Daphnia magna (p<0.001). Our findings indicated a considerable increase in DNA strand breakage in Daphnia magna after exposure to difenoconazole, indicating that the fungicide is genotoxic to daphnids.

Keywords

• Pesticide,
• Difenoconazole,
• Comet Assay
• DNA Damage

References

1. Ankley, G. T., & Villeneuve, D. L. (2006). The fathead minnow in aquatic toxicology: past, present and future. Aquatic toxicology, 78(1), 91-102.
2. Barillet, S., Adam-Guillermin, C., Palluel, O., Porcher, J. M., & Devaux, A. (2011). Uranium bioaccumulation and biological disorders induced in zebrafish (Danio rerio) after a depleted uranium waterborne exposure. Environmental Pollution, 159(2), 495-502.
3. Brunelli, E., Bernabò, I., Sperone, E., & Tripepi, S. (2010). Gill alterations as biomarkers of chronic exposure to endosulfan in Bufo bufo tadpoles. Histology and histopathology.
4. Cavalcante, D. G. S. M., Martinez, C. B. R., & Sofia, S. H. (2008). Genotoxic effects of Roundup® on the fish Prochilodus lineatus. Mutation Research/Genetic Toxicology and Environmental Mutagenesis, 655(1-2), 41-46.
5. Chen, X., Zheng, J., Zhang, J., Duan, M., Xu, H., Zhao, W., ... & Xu, Y. (2022). Exposure to difenoconazole induces reproductive toxicity in zebrafish by interfering with gamete maturation and reproductive behavior. Science of the Total Environment, 838, 155610.
6. Ergenler, A., & Turan, F. (2023). DNA Damage in Fish Due to Pesticide Pollution. Natural and Engineering Sciences, 8(3), 195-201.
7. European Food Safety Authority (EFSA), Álvarez, F., Arena, M., Auteri, D., Leite, S. B., Binaglia, M., ... and Villamar‐Bouza, L. (2024). Peer review of the pesticide risk assessment for the active substance difenoconazole in light of confirmatory data submitted. EFSA Journal, 22(7), e8921.
8. Felten, V., Toumi, H., Masfaraud, J. F., Billoir, E., Camara, B. I., & Férard, J. F. (2020). Microplastics enhance Daphnia magna sensitivity to the pyrethroid insecticide deltamethrin: Effects on life history traits. Science of the Total Environment, 714, 136567.

9. Gao, S., Jiang, J., Li, X., Ye, F., Fu, Y., & Zhao, L. (2021). An environmentally safe formulation with enhanced solubility and fungicidal activity: Self-assembly and characterization of Difenoconazole-β-CD inclusion complex. Journal of Molecular Liquids, 327, 114874.
10. Jiang, J., Chen, L., Wu, S., Lv, L., Liu, X., Wang, Q., & Zhao, X. (2020). Effects of difenoconazole on hepatotoxicity, lipid metabolism and gut microbiota in zebrafish (Danio rerio). Environmental Pollution, 265, 114844.
11. Knudsen, J. J., Holbech, H., Madsen, S. S., & Bjerregaard, P. (2011). Uptake of 17β-estradiol and biomarker responses in brown trout (Salmo trutta) exposed to pulses. Environmental pollution, 159(12), 3374-3380.
12. Li, X., Bai, Y., Zhu, W., Shi, X., & Xu, S. (2022). The endoplasmic reticulum-mitochondrial crosstalk is involved in the mitigation mechanism of eucalyptol on imidacloprid toxicity in Ctenopharyngodon idellus kidney cells. Fish & shellfish immunology, 127, 99-108.
13. Moreira, R. A., Daam, M. A., Vieira, B. H., Sanches, A. L. M., Reghini, M. V., da Silva Mansano, A., ... & Rocha, O. (2017). Toxicity of abamectin and difenoconazole mixtures to a Neotropical cladoceran after simulated run-off and spray drift exposure. Aquatic Toxicology, 185, 58-66.
14. Mu, X., Chai, T., Wang, K., Zhang, J., Zhu, L., Li, X., & Wang, C. (2015). Occurrence and origin of sensitivity toward difenoconazole in zebrafish (Danio reio) during different life stages. Aquatic toxicology, 160, 57-68.
15. Mu, X., Chai, T., Wang, K., Zhu, L., Huang, Y., Shen, G., ... & Wang, C. (2016). The developmental effect of difenoconazole on zebrafish embryos: A mechanism research. Environmental pollution, 212, 18-26.
16. Mu, X., Pang, S., Sun, X., Gao, J., Chen, J., Chen, X., ... & Wang, C. (2013). Evaluation of acute and developmental effects of difenoconazole via multiple stage zebrafish assays. Environmental Pollution, 175, 147-157.
17. Na, M. J., Lee, W. Y., & Park, H. J. (2024). Difenoconazole Induced Damage of Bovine Mammary Epithelial Cells via ER Stress and Inflammatory Response. Cells, 13(20), 1715.
18. Norusis, M. J. (1993). Advanced statistics. SPSS for Windows, Release 6.0, 578.
19. OECD. (1994). OECD Guidelines for the Testing of Chemicals. Oecd.
20. OECD Organisation for Economic Co-operation and Development (2012). Test Guideline No. 211: Daphnia Magna Reproduction Test. In: Organisation for Economic Co-operation and Development Paris, France.
21. OECD Organisation for Economic Co-operation and Development (2012). Test Guideline No. 211: Daphnia Magna Reproduction Test. In: Organisation for Economic Co-operation and Development Paris, France.
22. Strähle, U., Scholz, S., Geisler, R., Greiner, P., Hollert, H., Rastegar, S., ... & Braunbeck, T. (2012). Zebrafish embryos as an alternative to animal experiments—A commentary on the definition of the onset of protected life stages in animal welfare regulations. Reproductive Toxicology, 33(2), 128-132.
23. Tekin, Z., Ergenler, A., Zaman, B. T., Tahir Günkara, Ö., Depci, T., Turan, F., & Bakırdere, S. (2024). Microwave‐Assisted Synthesis Method for the Production of Carbon Nano Dots and Its Genotoxicity Assessment. ChemistrySelect, 9(37), e202403483.
24. Teng, M., Qi, S., Zhu, W., Wang, Y., Wang, D., Dong, K., & Wang, C. (2018). Effects of the bioconcentration and parental transfer of environmentally relevant concentrations of difenoconazole on endocrine disruption in zebrafish (Danio rerio). Environmental Pollution, 233, 208-217.
25. Ton, S. S., Chang, S. H., Hsu, L. Y., Wang, M. H., & Wang, K. S. (2012). Evaluation of acute toxicity and teratogenic effects of disinfectants by Daphnia magna embryo assay. Environmental pollution, 168, 54-61.
26. Turan, F., & Ergenler, A. (2022). Investigation of the genotoxic effect of acetamiprid in Cyprinus carpio using the micronucleus analysis and the comet assay. Turkish Journal of Maritime and Marine Sciences, 8(2), 80-89.
27. Turan, F., & Ergenler, A. (2023). The Genotoxic Damage in Cyprinus carpio Exposed to Abamectin. Natural and Engineering Sciences, 8(2), 119-128.
28. Wang, J., Gao, X., Liu, F., Dong, J., & Zhao, P. (2022). Difenoconazole causes cardiotoxicity in common carp (Cyprinus carpio): Involvement of oxidative stress, inflammation, apoptosis and autophagy. Chemosphere, 306, 135562.
29. Wang, K., Wu, J. X., & Zhang, H. Y. (2012). Dissipation of difenoconazole in rice, paddy soil, and paddy water under field conditions. Ecotoxicology and Environmental Safety, 86, 111-115.