Evaluation and Detection of Ochratoxins and Aflatoxins of Aspergillus piperis by Fluorometric Spectroscopy, Gel Electrophoresis and Western Blotting Techniques

Abstract

Total ochratoxins (OTs) and aflatoxins (AFs) of Aspergillus piperis were quantitively detected by fluorometric spectroscopy in both fungal biomass (FB) and culture filtrate (CF). OTs were detected with high concentrations; 38˟10-3 and 32˟10-3 μg/g in fungal biomass and culture filtrate of A. piperis respectively, while, AFs recorded lower concentrations; 3.3˟10-3 and 2.3˟10-3 μg/g. Consequently, different types of OTs produced in FB and CF of A. piperis were identified using sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) method. The molecular weights (MWs) of protein bands from SDS-PAGE step revealed the presence of ochratoxin B, ochratoxin α and ochratoxin C (OTB, OTα and OTC) in FB of A. piperis but, the results revealed that, only OTB and OTα were found in the CF. The MWs of OTB, OTα and OTC were found to be 363, 260,430 KDa respectively. Also, these findings were confirmed by western blotting technique using the antibodies of OTB, OTα and OTC. Toxicity of different concentrations of CF of A. piperis was tested in vitro on seed germination of broad bean (Vicia faba).