Azadirachtin’in Galleria mellonella Lepidoptera: Pyralidae üzerindeki genotoksisitesi

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

Anahtar Kelimeler:

azadirachtin, COMET, Galleria mellonella, genotoksisite, hemosit

Genotoxicity of azadirachtin on Galleria mellonella L. Lepidoptera: Pyralidae

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,

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