Molecular and Microscopic Detection of Microsporidia in Some

Year 2014, Volume: 1 Issue: 4, 450 - 453, 28.10.2014

Olgay Tekın Duran Sezer Kubra Sogancı Rahsan Ivgın Tunca

Abstract

 

The silkworm, an easy to grow and quite precious insect, is affected by many diseases like other organisms. The microsporidia spores caused to Pebrine which is quite dangerous and causes serious economic damage. In this study, 450 silkworm eggs (Bombyx mori L.) collected from Bursa province in Turkey were fed into ten different groups until the 4th and 5th larval stages. Nine groups were fed with unwashed mulberry leaves whereas only one group was fed with washed ones. The samples at the 4th and 5th larval stages from each group were screened for N. bombycis and Vairimorpha sp. using both microscopic and molecular methods. There were no nosema spores detected in dyed samples under microscope (100Xobjective). The primers, specific for N. bombycis NBEF35F, NBEF957R and Vairimorpha sp. M1196F, M11822R classified into microsporidia, were used for detection of disease from isolated DNAs. There was no pathogens detected using molecular technique. In conclusion, considering that infection is due to horizontal transmission, the study results may demonstrate that food materials and silkworm from breeding region are safe for disease pathogens. This study is the first molecular application for detection of N. bombycis and Vairimorpha sp. in Turkey.

Keywords

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Türkiye’de Bazı İpek Böceği (Bombyx mori L.) Populasyonlarında Microsporadianın Moleküler ve Mikroskobik Tespiti

Abstract

 

Yetiştirilmesi kolay ve oldukça kıymetli olan ipek böceği, diğer canlılarda da olduğu gibi birçok hastalıktan etkilenmektedir. Bunlardan birisi microsporadia sporlarının neden olduğu oldukça tehlikeli ve ciddi anlamda ekonomik zarara neden olan Pebrine hastalığıdır. Bu çalışmada, Bursa ilinden toplanan 450 adet ipek böceği (Bombyx mori L.) yumurtaları 4. ve 5. larval döneme kadar 10 farklı gruba ayrılarak beslenmiştir. İlk 9 grup yıkanmamış dut yaprakları ile beslenirken 1 grup yıkanmış dut yaprakları ile beslenmiştir. 4. ve 5. larval dönemdeki örnekler N. bombycis ve Vairimorpha sp. tespiti için mikroskobik ve moleküler metotlar kullanılarak taranmıştır. Boyanmış örnekler mikroskop (100Xbüyütme) altında incelendiğinde nosema sporuna rastlanmamıştır. İzole edilmiş DNA örneklerinde Nosema bombycis’e özgü NBEF35F, NBEF957R ve aynı zamanda microsporidia içinde yer alan Vairimorpha sp. M1196F, M11822R primerleri kullanılarak hastalık taraması yapılmıştır ve hastalık patojenine rastlanmamıştır. Sonuç olarak, horizontal bulaşma göz önüne alındığında, çalışma yetiştirmenin yapıldığı bölgenin ve yiyecek materyallerinin patojen bakımından güvenli olduğunu göstermiştir.

Keywords

Microsporidia, Vairimorpha, Nosema bombycis, PZR, İpekböceği (Bombyx mori L.)

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