Ispitivanje toksigenog potencijala gljiva primenom jednostavnog trijažnog metoda

Potential for the biosynthesis of aflatoxin B1 (AFLB1), ochratoxin A (OTA), diacetoxyscirpenol (DAS), T-2 toxin (T2), and zearalenone (ZON) was investigated in different fungal species belonging to the genera: Aspergillus, Fusarium and Penicillium. The majority of investigated isolates originated from cereal grains, crushed oil soybean seed and fodder mixtures. The simple screening method developed by Filtenborg et al. (1983) was applied with few modifications concerning the type of the medium and cultivation temperature. In order to optimise the biosynthetic conditions for different mycotoxins, the following control cultures, known as mycotin producers were used: OTA - A. ochraceus CBS 108.08, DAS - F. semitectum (SL-B i SL-C), T2 - F. sporotrichioides (ITM-391, M-1-1, R-2301) and ZON - F. graminearum (GZ-LES). The fungi were cultivated on the standard medium (YESA - 2% yeast extract, 15% sucrose and 2% agar, pH 6.5), three modifications of the basic medium (YESAZn - the standard medium supplemented with 0.23 mg/l ZnSO4 x 5 H2O; PPSA - the medium in which yeast extract was replaced with peptone-1; PPSAZn - the medium in which yeast extract was replaced with peptone-1 and supplemented with 0.23 mg/l ZnSO4 x 5 H2O), and the potato-dextrose agar (PDA). The earlier biosynthesis of tested mycotoxins was recorded under the following cultivation conditions of fungal species: AFLB1 - after 14 days on PDA at 27±1°C, OTA - after 10 days on YESA and YESAZn at 27±1°C, DAS - after 10 days on PPSA and PPSAZn at 27±1°C, T2 - after 7 days on PPSAZn and PPSA at room temperature (20-24°C), and ZON - after 1 week on YESA and YESAZn at room temperature (21-24°C).Potencijal za biosintezu aflatoksina B1 (AFLB1), ohratoksina A (OTA), diacetoksiscirpenola (DAS), T-2 toksina (T2) i zearalenona je ispitan kod izolata gljiva koje pripadaju rodovima Aspergillus, Fusarium i Penicillium. Izolati su uglavnom bili poreklom sa zrna žitarica i stočne hrane. Primenjena je jednostavna trijažna metoda F i l t e n b o r g-a i sar. (1983) kod koje su izvršene izvesne modifikacije u smislu tipa podloge i temperature kultivacije gljiva. U cilju optimizacije uslova za testiranje toksigenog profila odabranih gljivičnih izolata upotrebljene su kontrolne kulture za koje je prethodno dokazano da su proizvođači sledećih mikotoksina: OTA-a A. ochraceus CBS 108.08, DAS-a F. semitectum (SL-B i SL-C), T2 - F. sporotrichioides (ITM-391, M-1-1, R-2301) i ZON-a F. graminearum (GZ-LES). Gljive su gajene na standardnoj podlozi (EKSA - 2% ekstrakta kvasca i 15% saharoze, pH 6,5), tri modifikacije osnovne podloge (EKSAZn-standardna podloga sa dodatkom 0,23 mg/l ZnSO4 x 5 H2O; PPSA - podloga u kojoj je ekstrakt kvasca zamenjen peptonom-1; PPSAZn - podloga u kojoj je ekstrakt kvasca zamenjen peptonom-1 i kojoj je dodato 0,23 mg/l ZnSO4 x 5 H2O; pH 6,5) i krompir dekstroznoj podlozi (pH 6,5). Biosinteza ispitanih mikotoksina je najranije konstatovana pri sledećim uslovima gajenja gljiva: AFLB1 - posle 14 dana kultivacije na KDA i 27±1°C, OTA - posle 10 dana kultivacije na EKSA i EKSAZn i 27±1°C, DAS-a posle 10 dana kultivacije na PPSA i PPSAZn na 27±1°C, T2 - posle 7 dana na PPSA i PPSAZn i sobnoj temperaturi (20-24°C), i ZON-a posle nedelju dana na EKSA i EKSAZn i sobnoj temperaturi (21-24°C)