Biodiversity of Airborne Fungi in the Indoor Environment of Refrigerators Used in Houses

Year 2017, Volume: 8 Issue: 2, 109 - 124, 31.10.2017

Ahmet Asan , Soner Ozdil Burhan Sen Suzan Okten

Abstract

In this study, it was aimed to determination of microbial diversity from indoor air flora of refrigerators used at four different homes with conventional and molecular identifications. Research materials were obtained by taking samples twice each months from all refrigerators (totally twenty four times) at first and last weeks of October, November and December months of 2012. Air sampler (Millipore) were used to take samples and 100 L air was aspirated for each sampling. Dichloran Glycerol Agar (DG 18) medium was used on the sampling process. All isolates taken like a pure culture and to incubated at 25°C during 7 days after to passaged on slant Potato Dextrose Agar (PDA) medium. After that, these pure cultures at PDA medium were stored at 4°C as storage cultures. Conventional identifications of obtained isolates were diagnosed according to their features by using related monographs after to cultured on PDA, Malt Extract Agar (MEA), Czapek Yeast Extract Agar (CYA), Czapek Yeast Extract Agar with 20% Sucrose (CY20S), Glycerol Nitrate Agar (G25N), Czapeks-Dox Agar (CZ), Creatine Sucrose Agar (CREA), Yeast Extract Sucrose Agar (YES) and DG18  mediums. On the molecular identification, sequence analysis of obtained fungal amplicons were determined on Sanger Method at ABI Prism 377 DNA Sequencer (Applied Biosystems, ABD)  by using ABI prism Big Dye Terminator Cycle Sequencing Ready Reaction Kit and to compared with similar sequences on gene bank by BLAST Analysing. As a result of this study, it was identified what 34 % Penicillium, 24 % Cladosporium, 17 % Alternaria, 11 % Aspergillus species and 9 % other species as a percentage distribution of obtained fungi isolates.

Keywords

Airborne fungus, ITS gene region, Refrigerator, RT-Q-PCR, Sanger Sequencing

Evlerde Kullanılan Buzdolaplarının İç Ortamında Havayla Taşınan Fungusların Biyoçeşitliliği

Abstract

Bu çalışmada, 4 farklı evde kullanılan buzdolaplarının iç ortam havasında bulunan mikrofungusların morfolojik ve moleküler identifikasyonu yapılarak, buzdolabı hava ortamındaki mikrofungal çeşitliliğin tesbiti 

amaçlanmıştır. Araştırma materyali Ekim, Kasım ve Aralık 2012 aylarının ilk ve son haftasında 4 farklı istasyondan (her ay ikişer kez olmak üzere) toplam 24 kez örnekleme yapılarak elde edilmiştir. Hava örnekleme cihazı (Millipore) ile, bir örneklemede 100 L hava aspire edilerek alınmıştır. Örnekleme işleminde Dichloran Glycerol Agar (DG 18) besiyerleri kullanılmıştır. İzolatlar saf kültür olarak elde edilip yatık Potato Dextrose Agar (PDA)  besiyerine pasaj alınarak 25°C'de 7 gün inkübe edilmiş ve daha sonra stok kültür olarak 4°C'de saklanmıştır. Elde edilen izolatlar özelliklerine göre PDA, Malt Extract Agar (MEA), Czapek Yeast Extract Agar (CYA), Czapek Yeast Extract Agar with 20% Sucrose (CY20S), Glycerol Nitrate Agar (G25N), Czapeks-Dox Agar (CZ), Creatine Sucrose Agar (CREA), Yeast Extract Sucrose Agar (YES) ve DG 18 besiyerlerine ekilerek morfolojik teşhisleri ilgili monograflardan yararlanılarak yapılmıştır. Moleküler teşhiste ise elde edilen fungal amplikonların dizi analizleri Sanger yöntemiyle, ABI prism Big Dye Terminator Cycle Sequencing Ready Reaction Kit kullanılarak, ABI Prism 377 DNA Sequencer'da (Applied Biosystems, ABD) belirlenmiştir ve gen bankasındaki benzer sekanslarla BLAST Analizi yapılarak kıyaslanmıştır. Araştırmanın sonucunda teşhisi yapılan mantarların yüzdelik dağılımları % 34 Penicillium, % 24 Cladosporium, % 17 Alternaria, % 11 Aspergillus türleri ve % 9 diğer türler olarak tespit edilmiştir. 

Keywords

Havayla taşınan funguslar, ITS gen dizisi, Buzdolabı, RT-Q-PCR, Sanger Dizileme

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