Distribution of nontoxigenic strains of Aspergillus flavus throughout pistachio growing areas in Iran

Aflatoxins are highly toxic secondary metabolites produced by various Aspergillus species in food and feed. Nontoxigenic strains of A. flavus have been used for reducing aflatoxin of several crops. To determine the frequency of atoxigenic A. flavus population, samples were collected from different pistachio producing areas. A serial dilution method and AFPA medium was used to isolate Aspergillus section Flavi isolates. The ability of the isolates to produce aflatoxins was assayed using fluorescence detection (FD) around fungal colonies under UV 366 nm and color changing of colony after ammonia vapor (AV) exposure in the primary screening. First screening stage was validated by thin layer chromatography (TLC) after rice flour inoculation. The species of A. flavus were identified morphologically and molecularly based on species-specific primers. Overall, 524 isolates out of 681 isolates were identified as A. flavus. 53A. flavus isolates from nut and 10 isolates from soil did not produce any aflatoxins based on the methods used. Comparison of cultural and analytical methods showed FD methods overestimated the frequency of nontoxigenic strains while AV in combination with coconut agar medium produced similar results compare to TLC assays. The AV method can be used in the most laboratories as a practical, cheap, and rapid tool. Further efforts for assessment of competition potential of nontoxigenic strains versus toxigenic strains will be useful