Gıda katkı maddesi ferrolaktat (e585) 'ın drosophila melanogaster' de yaptığı genotoksik etkilerin smart testi ve comet assay yöntemiyle belirlenmesi
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Dosyalar
Tarih
2014
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Yayıncı
Trakya Üniversitesi Fen Bilimleri Enstitüsü
Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
Bu çalışmada, 72±4 saatlik D.melonagaster transheterezigot (mwh/flr3) larvaların ferrolaktat' lı besiyerlere maruz bırakılarak, İn vivo bir test sitemi olan somatik mutasyon ve rekombinasyon test sistemi (SMART) ile metamorfoz geçiren ergin bireylerin kanat imajinal disk hücrelerinde nokta mutasyon, delesyon, ayrılmama ve rekombinasyon sonucu meydana gelen genetik değişikliklerin Graf vd. (1984) sınıflandırma tekniğine dayanılarak kanatlardaki mutant trikomların tespit edilmesi ve istatistiksel olarak mutajenik ve rekombinojenik etkisi olup olmadığı araştırılmış olup, sonuçta 0.005' lik ferrolaktat derişimi uygulamasının kontrol grubu verileri ile karşılaştırıldığında istatistiksel anlamda fark oluşmadığı, 0.015 ve 0.025' lik ferrolaktat derişimlerinin ise istatistiksel olarak anlamlı/pozitif fark oluşturduğu gözlenmiştir. Oregon R+ soyuna ait yabanıl tip Drosophila melanogaster' in 72±4 saatlik larvalarının 24 saat süreyle ferrolaktat' lı besiyerlere maruz bırakılması ve 96±4 saatlik larvalardaki hemositlerin Irving vd. (2005) geliştirdiği teknik yardımıyla izole edilmesi, Singh vd. (1988) DNA hasarının tespit edilmesinde kullandıkları Comet Assay testi ile DNA hasarının belirlenmesi amacıyla, kuyruk yoğunluğu, kuyruk momenti ve kuyruk uzunluğu ortalamaları parametrelerinin kullanılarak kontrol grubu ile karşılaştırılması yapılmış, çalışma sonucunda; 0.005 ve 0.015' lik ferrolaktat derişimlerinin kontrol grubu verileri ile karşılaştırıldığında istatistiksel anlamda fark oluşmadığı, 0.025' lik ferrolaktat derişiminin ise istatistiksel olarak pozitif fark oluşturduğu gözlenmiştir. Sonuç olarak bu çalışmada, Drosophila melanogaster' de Kanat benek testi yöntemi (SMART) uygulaması ve Drosophila melanogaster hemositlerinde Comet Assay yöntemi ile yapılan uygulama sonuçlarında ferrolaktat' ın artan derişimlerinin DNA hasarına neden olduğu ve genotoksik risk oluşturabileceği tespit edilmiştir.
Abstract
In this study, three different concentrations 0.005, 0.015 and 0.025 of the food additive ferrous lactate (E585) were experimentally applied to Drosophila melanogaster larvae which were reared in the laborataory. The wing spot test, namely SMART (Somatic Recombination and Mutataion Test) and in Vivo Comet Assay tests were applied to analyse whether this food colouring agent has genotoxic effects in the larvae haemocytes of D. melanogaster. Three different concenntrations of ferroous lactate were added to the mediums where the flies were fed chronically during the course of the study. Also, negative control mediums prepared with distilled water and positive control mediums prepared with 4mM EMS (Ethyl Methyl Sülfonate) were used for the comparisons. The Kolmogorov-Smirnov test used to check the normality whether the data is appropriate for the parametric tests. The t-test was applied to see differences between the data groups wherever possible to test the statistical significance beyond the 0.05 level. According to SMART test results, the 0.005 concentrations of ferrous lactate did not show significant effect for inducing the mutant wing spots compared to control groups. However, the 0.015 and 0.025 concentrations of this food colouring agent induced significantly the number of mutant spots. The Comet Assay results also gave similar results. That is, the 0.005 concentration of ferrous lactate did not show significant effect for inducing the DNA damage in the larvae hemocytes compared to control groups. However, the other two concentrations of ferrous lactate signicicantly induced the damage associated with intensive, longer DNA tails and moments. Although as the concentraions of the food additive ferrous lactate increase in the medium it results more genotoxic effects, the significant genotoxic effects appear after about the 0.015 concentration in the larvae haemocytes of D. melanogaster.
Abstract
In this study, three different concentrations 0.005, 0.015 and 0.025 of the food additive ferrous lactate (E585) were experimentally applied to Drosophila melanogaster larvae which were reared in the laborataory. The wing spot test, namely SMART (Somatic Recombination and Mutataion Test) and in Vivo Comet Assay tests were applied to analyse whether this food colouring agent has genotoxic effects in the larvae haemocytes of D. melanogaster. Three different concenntrations of ferroous lactate were added to the mediums where the flies were fed chronically during the course of the study. Also, negative control mediums prepared with distilled water and positive control mediums prepared with 4mM EMS (Ethyl Methyl Sülfonate) were used for the comparisons. The Kolmogorov-Smirnov test used to check the normality whether the data is appropriate for the parametric tests. The t-test was applied to see differences between the data groups wherever possible to test the statistical significance beyond the 0.05 level. According to SMART test results, the 0.005 concentrations of ferrous lactate did not show significant effect for inducing the mutant wing spots compared to control groups. However, the 0.015 and 0.025 concentrations of this food colouring agent induced significantly the number of mutant spots. The Comet Assay results also gave similar results. That is, the 0.005 concentration of ferrous lactate did not show significant effect for inducing the DNA damage in the larvae hemocytes compared to control groups. However, the other two concentrations of ferrous lactate signicicantly induced the damage associated with intensive, longer DNA tails and moments. Although as the concentraions of the food additive ferrous lactate increase in the medium it results more genotoxic effects, the significant genotoxic effects appear after about the 0.015 concentration in the larvae haemocytes of D. melanogaster.
Açıklama
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Anahtar Kelimeler
Ferrolaktat, Genotoksisite, Somatik Rekombinasyon ve Mutasyon Testi (SMART), Drosophila Melanogaster, Comet Assay, Somatic Mutation and Recombination Test (SMART), Genotoxicity, Drosophila Melanogaster, Ferrouslactate, Comet Assay
