title: Characterisation of the yeast and mould biota in traditional white pickled cheeses by culture-dependent and independent molecular techniques.
abstract: Artisanal white pickled cheese of Western Serbia is a product of complex microbial community which detection by culture-dependent method only is hampered by its limitations. Thus, in the present study, we used a culture-independent, semi-quantitative technique based on construction of an internal transcribed spacer (ITS)-clone library from metagenomic DNA. This approach, based on direct DNA extraction followed by amplification of fungal internal transcribed regions (ITS) cloned into plasmid and restricted by endonucleases, revealed greater species richness in analysed cheeses and their by-products (17 species in total) compared to the more commonly used techniques of the culture-dependent method (8 species) and LSU-DGGE (10 species). The most frequently occurring yeast species which are commonly associated with cheeses production were Debaryomyces hansenii, Kluyveromyces lactis and Candida zeylanoides. On the other hand, Yarrowia lipolytica and Galactomyces geotrichum were detected only in one cheese sample. Moreover, some species, mainly moulds (Filobasidium globisporum, Cladosporium sp., Aspergillus sp. or Alternaria sp.) were identified only by culture-independent methods. The discrepancies between the techniques were confirmed by low correlation factor and by different indices of general biodiversity and dominance of species. The ITS-clone library approach provides the opportunity to analyse complex fungal communities associated with food products.
authors.affiliation: Biotechnical Faculty, University of Ljubljana, Jamnikarjeva 101, 1000, Ljubljana, Slovenia.|Institute of Food Science and Biotechnology, Faculty of Chemistry, Brno University of Technology, Brno, Czech Republic.
authors.forename: Hana
authors.initials: H
authors.lastname: Šuranská
authors.affiliation: Biotechnical Faculty, University of Ljubljana, Jamnikarjeva 101, 1000, Ljubljana, Slovenia.
authors.forename: Peter
authors.initials: P
authors.lastname: Raspor
authors.affiliation: Faculty of Food Technology and Biotechnology, University of Zagreb, Zagreb, Croatia.
authors.forename: Ksenija
authors.initials: K
authors.lastname: Uroić
authors.affiliation: Institute of Molecular Genetics and Genetic Engineering, University of Belgrade, Belgrade, Serbia.
authors.forename: Nataša
authors.initials: N
authors.lastname: Golić
authors.affiliation: Faculty of Food Technology and Biotechnology, University of Zagreb, Zagreb, Croatia.
authors.forename: Blaženka
authors.initials: B
authors.lastname: Kos
authors.affiliation: Institute of Molecular Genetics and Genetic Engineering, University of Belgrade, Belgrade, Serbia.
authors.forename: Sanja
authors.initials: S
authors.lastname: Mihajlović
authors.affiliation: Institute of Molecular Genetics and Genetic Engineering, University of Belgrade, Belgrade, Serbia.
authors.forename: Jelena
authors.initials: J
authors.lastname: Begović
authors.affiliation: Faculty of Food Technology and Biotechnology, University of Zagreb, Zagreb, Croatia.
authors.forename: Jagoda
authors.initials: J
authors.lastname: Šušković
authors.affiliation: Institute of Molecular Genetics and Genetic Engineering, University of Belgrade, Belgrade, Serbia.
authors.forename: Ljubiša
authors.initials: L
authors.lastname: Topisirović
authors.affiliation: Biotechnical Faculty, University of Ljubljana, Jamnikarjeva 101, 1000, Ljubljana, Slovenia. neza.cadez@bf.uni-lj.si.
authors.forename: Neža
authors.initials: N
authors.lastname: Čadež
mesh_terms: D044822:Biodiversity; D002611:Cheese / Q000382:microbiology*; D004271:DNA, Fungal / Q000235:genetics; D021903:DNA, Ribosomal Spacer / Q000235:genetics; D005658:Fungi / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification*; D012150:Polymorphism, Restriction Fragment Length; D055771:Serbia
chemical_list: D004271:DNA, Fungal; D021903:DNA, Ribosomal Spacer