Phenotypic differentiation of species from Aspergillus section Flavi on neutral red desiccated coconut agar

In order to facilitate easy and rapid identification of aflatoxin-producing Aspergillus species, the phenotypic traits of Aspergillus section Flavi isolates were examined on neutral red desiccated coconut agar (NRDCA). Phenotype variations in colony morphology and the relationship between colour/intensity of fluorescence and aflatoxin production were assessed. The isolates included 10 Aspergillus minisclerotigenes strains, 11 non-aflatoxigenic Aspergillus flavus L strains, 29 aflatoxigenic A. flavus L strains and 20 strains each of Aspergillus parasiticus and Aspergillus parvisclerotigenus. The NRDCA medium supported morphological differentiation of the four species based on colony features, conidia type and colour. In particular, the two very closely related minisclerotial species, A. minisclerotigenes and A. parvisclerotigenus, were clearly differentiated by their colony colour on NRDCA. All toxigenic isolates produced aflatoxins in the culture medium in varying quantities. Plates of aflatoxigenic A. flavus L strains fluoresced bluish purple/lavender around the colony on the obverse and pastel blue on the reverse side due to aflatoxin B production while those of A. minisclerotigenes, A. parasiticus and A. parvisclerotigenus fluoresced with a light blue or light turquoise ring around the colony on the obverse and light sky blue or cadet blue on the reverse side depending on the amount of aflatoxin B and G produced. The colour of fluorescence significantly correlated (r=0.95, P=0.001) with the type(s) of aflatoxins produced by the isolates. In addition, the concentration of aflatoxins significantly (r=0.92; P=0.001) influenced the intensity of fluorescence in the aflatoxin-producing species. NRDCA can therefore be used for the rapid identification of Aspergillus section Flavi species based on colonial characteristics, and grouping of species into B and B+G aflatoxin producers within 5 days thus obviating the need for chemical analysis of the culture.