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Highly toxigenic strains of Aspergillus flavus have been reported to frequently contaminate maize, causing fatal aflatoxin poisoning in Kenya. To gain insights into the environmental and genetic factors that influence toxigenicity, fungi (n = 218) that were culturally identified as A. flavus were isolated from maize grains samples (n = 120) from three regions of Kenya. The fungi were further characterized to confirm their identities using a PCR-sequence analysis of the internal transcribed spacer (ITS) region of rDNA which also revealed all of them to be A. flavus. A subset of 72 isolates representing ITS sequence-based phylogeny cluster and the agroecological origin of maize samples was constituted for subsequent analysis. The analysis of partial calmodulin gene sequences showed that the subset consisted of A. flavus (87%) and Aspergillus minisclerotigenes (13%). No obvious association was detected between the presence of seven aflatoxin biosynthesis genes and fungal species or region. However, the presence of the aflD and aflS genes showed some association with aflatoxin production. The assessment of toxigenicity showed higher aflatoxin production potential in A. minisclerotigenes isolates. Given that A. minisclerotigenes were mainly observed in maize samples from Eastern Kenya, a known aflatoxin hotspot, we speculate that production of copious aflatoxin is an adaptative trait of this recently discovered species in the region.

Filamentous fungi were isolated from ten spices collected from markets in Indonesia. The aim was to enumerate fungal contamination and to determine the toxigenicity of Aspergillus flavus strains on each of the spices. Viable fungal populations were determined using a dilution method. Toxigenicity of Aspergillus flavus was determined by culture on a quick screening coconut agar and by PCR using four sets of primers specific for aflatoxin pathway genes. All the tested spices were contaminated by storage fungi, Species of Aspergillus was the most commonly isolated moulds followed by species of Fusarium, Mucor, Penicillium and Rhizopus. The greatest number of Aspergillus flavus isolates were found on white pepper, followed by nutmeg, cardamom, and black pepper. The greatest number of Aspergillus chevalieri isolates were found on coriander, followed by nutmeg. Fifty strains of A. flavus were isolated, all of the strains produced large sclerotia and biseriate conidiophores. Their toxigenicity was assayed by the presence of yellow pigment on a quick screening coconut agar medium and PCR amplification of regulatory and structural genes in the aflatoxin pathway.

Aflatoxins contaminate foodstuff posing a severe threat to human health because chronic exposure is linked to liver cancer while acute exposure may cause death. Therefore, it is of interest to reduce the contamination of crops by aflatoxins in the field and post-harvest. Among the current technologies being developed is the deployment of non-aflatoxigenic strains of Aspergillus species to competitively exclude aflatoxigenic conspecifics from crops in the field thereby curtailing aflatoxin production by the former. The success in this endeavor makes the non-aflatoxigenic fungi good candidates for biological control programs. However, the current techniques for segregating non-aflatoxigenic from aflatoxigenic fungi suffer two main drawbacks: they are based on morphological and chemical tests with a combination of visual color changes detected in a culture plate which suffer some degree of inaccuracy. Secondly, the existing methods are incapable of accurately quantifying aflatoxin production by fungi in culture. We developed a culture system for inducing aflatoxin production by Aspergillus using maize kernels as growth substrate followed by quantification using ELISA. The method was compared to the Dichlorvos-Ammonia (DV-AM) method for determining aflatoxigenicity. Our findings encapsulate a method more robust than the currently used DV-AM approach because, for the first time, we are able to assess aflatoxigenicity and aflatoxigenic variability among Aspergillus species earlier classified as non-aflatoxigenic by the DV-AM method. Furthermore, the new method presents an opportunity to attribute toxin production by actively growing fungal cultures. We believe this method when further developed presents a chance to study and predict fungal behavior prior to field trials for biological control programs.