The biochemical profile of Aspergillus niger environmental isolates was obtained using the Biolog MicroStation test system (MicroLog3 ver. 4.20.05, Biolog Inc., United States). By using a 96-well Biolog FF MicroPlate, unique phenotypic fingerprinting can be obtained for individual strains; this is a manifestation of their catabolic potential. Metabolic reactions, dependent on the production of suitable enzymes for the oxidation of the tested substrates, are quantified by the formation of tetrazolium dye (Bochner, 2009). These results, as fingerprint reaction patterns, provide a lot of information about the individual differences in the metabolic properties of each fungus tested.

All A. niger environmental isolates were cultured in tubes on 2% slants of MEA (Malt Extract Agar HiMedia Laboratories, Mumbai, India) at 26°C for 5–7 days. Conidia were then inoculated into FF-IF inoculating fluid (72106 FF-IF inoculating fluid, Biolog, United States) according to the Biolog protocol and the density was adjusted to the recommended value of 65%, which was measured using a turbidimeter. Subsequently, we inoculated 100 μL of conidia suspension onto plates containing a pre-set of test substrates. We incubated the microplates for 72–168 h and then evaluated them using the Biolog MicroStation system.

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