Genotoxicity of azadirachtin on Galleria mellonella L. Lepidoptera: Pyralidae

Year 2018, Volume: 11 Issue: 3, 24 - 30, 15.12.2018

Emine Duman Hülya Altuntaş

https://izlik.org/JA26EC83UU

Abstract

In this paper, genotoxic effects of the pure azadirachtin AZA on the larval hemocytes of model insect and storage pest Galleria mellonella L. Lepidoptera: Pyralidae was investigated. The comet assay was performed to measure and analyze DNA damage in larval hemocytes at Anadolu University Eskisehir Technical University between 2017 and 2018 years. For this purpose, sublethal AZA doses 0.5, 1, 1.5, and 2 μg/larva given to G. mellonella larvae via insect force feeding method were used to monitor tail intensity, tail moment and tail migration the commonly known comet parameters. These DNA damage indicators were analyzed in hemocytes obtained from larvae at 24 and 72 h post force feeding. All comet parameters at all doses of AZA increased in comparison with negative and positive control at 24 and 72 h. At 72 h post force feeding with median lethal dose of AZA, a significant increase in DNA damage indicators was observed in larval hemocytes as compared with untreated groups. Consequently, this study showed that AZA caused significant damage in the genome of G. mellonella larvae even at sublethal doses and the comet assay was useful in the monitoring of in vivo genotoxicity of AZA in larva hemocytes

Keywords

azadirachtin , COMET , Galleria mellonella , genotoxicity , hemocytes

Azadirachtin’in Galleria mellonella Lepidoptera: Pyralidae üzerindeki genotoksisitesi

https://izlik.org/JA26EC83UU

Abstract

Bu çalışmada, azadirachtin AZA ’in model böcek ve depo zararlısı olan Galleria mellonella L. Lepidoptera: Pyralidae ’nın larval hemositleri üzerindeki genotoksik etkileri araştırılmıştır. Larval hemositlerdeki DNA hasarının analiz edilmesi ve ölçülmesi comet analizi kullanılarak, 2017-2018 yılları arasında Anadolu Üniversitesi Eskişehir Teknik Üniversitesi ’nde gerçekleştilmiştir. Bu amaçla, hemositlerdeki kuyruk yoğunluğu, kuyruk momenti ve kuyruk göçü gibi önemli comet parametrelerinin analiz edilmesi için G. mellonella larvalarına, böcekler için ağızdan besleme yöntemiyle verilen subletal AZA dozları 0.5, 1, 1.5 ve 2 μg/larva kullanılmıştır. Ağızdan besleme uygulamasından 24 ve 72 saat sonra larvalardan elde edilen hemositlerde bu DNA indikatörleri analiz edilmiştir. AZA uygulanmasından 24 ve 72 saat sonra, larval hemositlerde belirlenen tüm comet parametrelerinde negatif ve pozitif kontrol grubuna kıyasla hem doza hem de zamana bağlı olarak artış meydana geldiği görülmüştür. Özellikle, ortalama letal doz AZA uygulamasından 72 saat sonra, larval hemositlerdeki DNA hasarı indikatörlerinde kontrol gruplarına kıyasla önemli bir artış gözlenmiştir. Sonuç olarak, bu çalışma AZA’nın subletal dozlarda bile G. mellonella larval genomunda önemli oranda hasara neden olduğunu ve larval hemositlerinde AZA’nın genotoksisitesinin in vivo değerlendirilmesinde comet analizinin kullanışlı olduğunu göstermiştir

Keywords

azadirachtin , COMET , Galleria mellonella , genotoksisite , hemosit

References

• Altuntaş, H., Demirci, S. N. Ş., Duman, E., Ergin, E. (2016). Toxicological and physiological effects of ethephon on the model organism, Galleria mellonella L. 1758 (Lepidoptera: Pyralidae). Türkiye Entomoloji Dergisi, 40(4), 413-423.
• Altuntaş, H., Duman, E. (2017). Toxicological effects of the entomopathogenic Purpureocillium lilacinus on the model organism Galleria mellonella. Biological Diversity and Conservation, 10(1), 153-159.
• Anderson, D., Dobrzynka, M. M., Jackson, L. I., Yu T. W., Brinkworth, M. H. (1997). Somatic and germ cell effects in rats and mice after treatment with 1,3-butadiene and its metabolites, 1,2-epoxybutane and 1,2,3,4-diepoxybutane. Mutation Research, 391, 233-242.
• Aribi, N., Oulhaci, M. C., Kilani-Morakchi, S., Sandoz, J. C., Kaiser, L., Denis. B., Joly, D. (2017). Azadirachtin impact on mate choice, female sexual receptivity and male activity in Drosophila melanogaster (Diptera: Drosophilidae). Pesticide Biochemistry and Physiology, http://dx.doi.org/10.1016/j.pestbp.2017.09.002.
• Asaduzzaman, M., Shim, J. K., Lee, S., Lee, K. Y. (2016). Azadirachtin ingestion is lethal and inhibits expression of ferritin and thioredoxin peroxidase genes of the sweetpotato whitefly Bemisia tabaci. Journal of Asia-Pacific Entomology, 19, 1-4.
• Boulahbel, B., Aribi, N., Kilani-Morakchi, S., Soltani, N. (2015). Insecticidal activity of azadirachtin on Drosophila melanogaster and recovery of normal status by exogenous 20-hydroxyecdysone. African Entomology, 23, 224-233.
• Charriere, J. D., Imdorf, A. (1997). Protection of honey combs from moth damage. Swiss Bee Research Center Federal Dairy Research Station, CH-3003, Communication, Nr. 24, Liebefeld, Bern, Switzerland, 3-4.
• Collins, A. R., (2004). The comet assay for DNA damage and repair: principles, applications, and limitations. Molecular Biotechnology, 26, 249-261.
• Dere, B., H. Altuntaş, H., Nurullahoğlu, Z. U. (2015). Insecticidal and oxidative effects of azadirachtin on the model organism Galleria mellonella L. (Lepidoptera: Pyralidae). Archives of Insect Biochemistry and Physiology, 89, 138- 152.
• Er, A., Taşkıran, D., Sak, O. (2017). Azadirachtin-induced effects on various life history traits and cellular immune reactions of Galleria mellonella (Lepidoptera: Pyralidae). Arch Archives of Biological Sciences, 69(2), 335-344.
• Fairbairn, D. W., Walburger, D. K., Fairbairn J. J., O’Neill, K. L. (1996). Key morphologic changes and DNA strand breaks in human lymphoid cells: Discriminating apoptosis from necrosis. Scanning, 18, 407-416.
• Huang, J. F., Shui, K. J., Li, H. Y., Hu, M. Y., Zhong, G. H. (2011). Antiproliferative effect of azadirachtin on Spodoptera litura Sl-1 cell line through cell cycle arrest and apoptosis induced by upregulation of p53. Pesticide Biochemistry and Physiology, 99, 16-24.
• Knopper, L. D. (2005). Use of the Comet Assay to Assess Genotoxicity in Mammalian, Avian, and Amphibian Species. Canadian Wildlife Service, Technical Report Series, 429.
• Kwadha, C. A., Ong’amo, G. O., Ndegwa, P. N., Raina, S. K., Fombong, A. T. (2017). The Biology and Control of the Greater Wax Moth, Galleria mellonella. Insects, 8, 61.
• Maria-Packiam, S., Emmanuel, C., Baskar, K., Ignacimuthu, S. (2015). Feeding Deterrent and Genotoxicity Analysis of a novel Phytopesticide by using Comet Assay against Helicoverpa armigera (HÜbner) (Lepidoptera: Noctuidae). Archives of Biology and Technology, 58(4), 487-493.
• Martelli, A., Carrozzino, R., Mattioli, F., Bucci, G., Lamarino, G., Brambilla, G. (2002). DNA damage in tissues of rat treated with potassium canrenoate. Toxicology, 171, 95-103.
• Muangphra, P., Gooneratne, R. (2011). Toxicity of Commercial Neem Extract to Earth worms (Pheretima peguana). Corporation Applied and Environmental Soil Science, doi:10.1155/2011/925950.
• Mukhopadhyay, I., Chowdhuri, D. K., Bajpayee, M., Dhawan, A. (2004). Evaluation of in vivo genotoxicity of cypermethrin in Drosophila melanogaster using the alkaline Comet assay. Mutagenesis, 19(2), 85-90.
• Nicolopoulou-Stamati, P., Maipas, S., Kotampasi, C., Stamatis, P., Hens, L. (2016), Chemical pesticides and human health: the urgent need for a new conceptin agriculture. Front Public Health, 4, 148.
• Olive, P. L., Banáth, J. P. (2006). The comet assay: a method to measure DNA damage in individual cells. Nature Protocols, 1, 23-29.
• Olson, S. (2015). An analysis of the biopesticide market now and where it is going. Outlooks on Pest Management, 26, 203-206.
• Qari, S. H., Abdel-Fattah, N. A. H., Shehawy, A. A. (2017). Assessment of DNA Damage and Biochemical Responses in Rhyzopertha dominica Exposed to Some Plant Volatile Oils. Journal of Pharmacology and Toxicology, 12(2), 87- 96.
• Qiao, J., Zou, X., Lai, D., Yan, Y., Wang, Q., Li, W., Deng, S., Xu, H., Gu, H. (2013). Azadirachtin blocks the calcium channel and modulates the cholinergic miniature synaptic current in the central nervous system of Drosophila. Pest Management Science, 70, 1041-1047.
• Rajaguru, P., Vidya, L., Baskarasethupathi, B., Kumar, P. A., Palanivel, M., Kalaiselvi, K. (2002). Genotoxicity evaluation of polluted groundwater in human peripheral blood lymphocytes using the comet assay. Mutation Research, 517, 29-37.
• Rembold, H., Annadurai, R. S. (1993). Azadirachtin inhibits proliferation of Sf9 cells in monolayer culture. Z Naturforsch, 48, 495-499.
• Sanchez, L. E. C., Osornio, J. J. J., Herrera, M. A. D. (2010). Secondary metabolites of the annonaceae, solanaceae and meliaceae families used as biological control of insects. Tropical and Subtropical Agroecosystems, 12, 445-462.
• Saxena, A., Kesari, V. P. (2016). Lack of genotoxic potential of pesticides, spinosad, imidacloprid and neem oil in mice (Mus musculus). Journal of Environmental Biology, 37, 291-295.
• Shao, X., Lai, D., Zhang, L., Xu, H. (2016). Induction of autophagy and apoptosis via PI3K/ AKT/TOR pathways by Azadirachtin A in Spodoptera litura Cells. Scientific Reports, 6, 35482.
• Shu, B., Wang, W., Hu, Q., Huang, J., Hu, M., Zhong, G. (2015). A comprehensive study on apoptosis induction by azadirachtin in Spodoptera frugiperda cultured cell line Sf9. Archives of Insect Biochemistry and Physiology, 89, 153-168.
• 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, 184-191.
• Zhang, Y., Zhong, Y., Luo, Y., Kong, Z. M. (2000). Genotoxicity of two novel pesticides for the earthworm, Eisenia fetida. Environmental Pollution, 208(2), 271-278.