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

Year 2017, Volume: 8 Issue: 1, 68 - 75, 28.04.2017

Duygu Kadaifçiler , Zuhal Zeybek

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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