Floresanlı Yerinde Hibritleme ve Geleneksel Kültür Yöntemlerini Kullanılarak Soğutma Kulesi Örneklerinde Fungusların Tespiti

Abstract

Doğada ve insan yapımı su sistemlerinde yaygın olarak bulunan Eurotiales üyesi fungusların varlığının tespiti, fırsatçı enfeksiyon ve alerjilere yol açabilen bu funguslara karşı alınabilecek önlemler açısından önemlidir. Bu çalışmada soğutma kulesi örneklerindeki fungusların varlığı, hem floresanlı yerinde hibritleme (FISH) hem de kültür yöntemiyle araştırılmış, her iki yöntemin sonuçları karşılaştırılmıştır. Bu amaçla İstanbul’daki farklı soğutma kulelerine ait toplam 40 adet su ve biyofilm örnekleri alınarak incelenmiştir. Kültür yönteminde tüm örnekler, ön işlemlerden sonra izolasyon besiyerine ekilmiştir. Ekimlerin sonuçları 14 gün sonunda değerlendirilmiştir. FISH yönteminde ise; tüm örnekler fikse edilip uygun problar kullanılarak hibridizasyonu yapıldıktan sonra epifloresan mikroskopta incelenmiştir. FISH yöntemi ile 3 gün gibi kısa bir sürede sonuç elde edilmiştir. Ayrıca bu yöntem ile incelenen örneklerin tamamında, kültür yönteminde ise incelenen örneklerin %97.5’in de fungus tespit edilmiştir. Kültür yöntemi ile en çok izole edilen fungusların Aspergillus ve Penicillium cinsine ait oldukları belirlenmiştir. Çalışma sonucunda her iki yöntemin beraber kullanılması önerilmektedir.

Keywords

• Aspergillus,kültür bağımsız yöntem,Eurotiales,floresanlı yerinde hibritleme

Detection of Fungi in Cooling Tower Samples Using Fluorescent In Situ Hybridization and Traditional Culture-Dependent Methods

Abstract

The determination of the presence of Eurotiales members, which are widely distributed in nature and in man-made aquatic systems, is important for taking measures against those capable of creating opportunistic infections and allergies. In this study, the presence of fungi in cooling-tower samples was determined by using two different methods, one being fluorescent in situ hybridization (FISH), the second culturing; both sets of results were compared with each other. For this purpose, a total of 40 water and biofilm samples were taken from different cooling towers in Istanbul for investigation. In the culture method, all samples were applied to the isolation medium after pretreatment. The results of the applications were evaluated after 14 days. With the FISH method, all samples were fixed and hybridized by using suitable probes and were investigated under an epifluorescent microscope. Results were obtained after a short period of three days. The FISH method was able to detect 100% of fungi in all samples, whereas the culture method was able to detect 97.5% of the fungi in the samples. With culture method, the most isolated fungi were from the Aspergillus and Penicillium genera. As a result of the study, both methods are recommended to be used in conjunction.

Keywords

• Aspergillus,culture-independent method,Eurotiales,fluorescent in situ hybridization

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