Degeneration of aflatoxin gene clusters in Aspergillus flavus from Africa and North America

Open Access journal; Published online: 31 August 2016Aspergillus flavus is the most common causal agent of aflatoxin contamination of food and feed. However, aflatoxin-producing potential varies widely among A. flavus genotypes with many producing no aflatoxins. Some non-aflatoxigenic genotypes are used as biocontrol agents to prevent contamination. Aflatoxin biosynthesis genes are tightly clustered in a highly conserved order. Gene deletions and presence of single nucleotide polymorphisms (SNPs) in aflatoxin biosynthesis genes are often associated with A. flavus inability to produce aflatoxins. In order to identify mechanisms of non-aflatoxigenicity in non-aflatoxigenic genotypes of value in aflatoxin biocontrol, complete cluster sequences of 35 A. flavus genotypes from Africa and North America were analyzed. Inability of some genotypes to produce aflatoxin resulted from deletion of biosynthesis genes. In other genotypes, non-aflatoxigenicity originated from SNP formation. The process of degeneration differed across the gene cluster; genes involved in early biosynthesis stages were more likely to be deleted while genes involved in later stages displayed high frequencies of SNPs. Comparative analyses of aflatoxin gene clusters provides insight into the diversity of mechanisms of non-aflatoxigenicity in A. flavus genotypes used as biological control agents. The sequences provide resources for both diagnosis of non-aflatoxigenicity and monitoring of biocontrol genotypes during biopesticide manufacture and in the environment

Aspergillus section Flavi community structure in Zambia influences aflatoxin contamination of maize and groundnut

Article purchased; Published online: 19 August 2017Aspergillus section Flavi have been implicated as causal agents of aflatoxin contamination in Africa. However, Aspergillus populations associated with aflatoxin contamination in Zambia have not been adequately detailed. Most of Zambia's arable land is non-cultivated and Aspergillus communities in crops may originate in non-cultivated soil. However, relationships between Aspergillus populations on crops and those resident in non-cultivated soils have not been explored. Because characterization of similar fungal populations outside of Zambia have resulted in strategies to prevent aflatoxins, the current study sought to improve understanding of fungal communities in cultivated and non-cultivated soils and in crops. Crops (n = 412) and soils from cultivated (n = 160) and non-cultivated land (n = 60) were assayed for Aspergillus section Flavi from 2012 to 2016. The L-strain morphotype of Aspergillus flavus and A. parasiticus were dominant on maize and groundnut (60% and 42% of Aspergillus section Flavi, respectively). Incidences of A. flavus L-morphotype were negatively correlated with aflatoxin in groundnut (log y = 2.4990935 − 0.09966x, R2 = 0.79, P = 0.001) but not in maize. Incidences of A. parasiticus partially explained groundnut aflatoxin concentrations in all agroecologies and maize aflatoxin in agroecology III (log y = 0.1956034 + 0.510379x, R2 = 0.57, P < 0.001) supporting A. parasiticus as the dominant etiologic agent of aflatoxin contamination in Zambia. Communities in both non-cultivated and cultivated soils were dominated by A. parasiticus (69% and 58%, respectively). Aspergillus parasiticus from cultivated and non-cultivated land produced statistically similar concentrations of aflatoxins. Aflatoxin-producers causing contamination of crops in Zambia may be native and, originate from non-cultivated areas, and not be introduced with non-native crops such as maize and groundnut. Non-cultivated land may be an important reservoir from which aflatoxin-producers are repeatedly introduced to cultivated areas. The potential of atoxigenic members of the A. flavus-L morphotype for management of aflatoxin in Zambia is also suggested. Characterization of the causal agents of aflatoxin contamination in agroecologies across Zambia gives support for modifying fungal community structure to reduce the aflatoxin-producing potential

Influence of wounding and temperature on resistance of maize landraces from Mexico to aflatoxin contamination

Open Access journal; Published online: 24 Sept 2020Maize is a staple for billions across the globe. However, in tropical and sub-tropical regions, maize is frequently contaminated with aflatoxins by Aspergillus section Flavi fungi. There is an ongoing search for sources of aflatoxin resistance in maize to reduce continuous exposures of human populations to those dangerous mycotoxins. Large variability in susceptibility to aflatoxin contamination exists within maize germplasm. In Mexico, several maize landrace (MLR) accessions possess superior resistance to both Aspergillus infection and aflatoxin contamination but their mechanisms of resistance have not been reported. Influences of kernel integrity on resistance of four resistant and four susceptible MLR accessions were evaluated in laboratory assays. Wounds significantly (P 0.05) proportionally support higher aflatoxin accumulation suggesting differential influences by some resistance factors between sporulation and aflatoxin biosynthesis. Physical barriers (i.e., wax and cuticle) prevented both aflatoxin accumulation and A. flavus sporulation in a highly resistant MLR accession. In addition, influence of temperature on aflatoxin contamination was evaluated in both viable and non-viable kernels of a resistant and a susceptible MLR accession, and a commercial hybrid. Both temperature and living embryo status influenced (P < 0.05) resistance to both aflatoxin accumulation and A. flavus sporulation. Lower sporulation on MLR accessions suggests their utilization would result in reduced speed of propagation and associated epidemic increases in disease both in the field and throughout storage. Results from the current study should encourage researchers across the globe to exploit the large potential that MLRs offer to breed for aflatoxin resistant maize. Furthermore, the studies provide support to the importance of resistance based on the living host and maintaining living status to reducing episodes of post-harvest contamination

Prevalence and mitigation of aflatoxins in Kenya (1960-to date)

Article PurchasedAflatoxins are highly toxic metabolites of several Aspergillus species widely distributed throughout the environment. These toxins have adverse effects on humans and livestock at a few micrograms per kilogram (μg/kg) concentrations. Strict regulations on the concentrations of aflatoxins allowed in food and feed exist in many nations in the developing world. Loopholes in implementing regulations result in the consumption of dangerous concentrations of aflatoxins. In Kenya, where ‘farm-to-mouth’ crops become severely contaminated, solutions to the aflatoxins problem are needed. Across the decades, aflatoxins have repeatedly caused loss of human and animal life. A prerequisite to developing viable solutions for managing aflatoxins is understanding the geographical distribution and severity of food and feed contamination, and the impact on lives. This review discusses the scope of the aflatoxins problem and management efforts by various players in Kenya. Economic drivers likely to influence the choice of aflatoxins management options include historical adverse health effects on humans and animals, cost of intervention for mitigation of aflatoxins, knowledge about aflatoxins and their impact, incentives for aflatoxins safe food and intended scope of use of interventions. It also highlights knowledge gaps that can direct future management efforts. These include: sparse documented information on human exposure; few robust tools to accurately measure economic impact in widely unstructured value chains; lack of long-term impact studies on benefits of aflatoxins mitigation; inadequate sampling mechanisms in smallholder farms and grain holding stores/containers; overlooking social learning networks in technology uptake and lack of in-depth studies on an array of aflatoxins control measures followed in households. The review proposes improved linkages between agriculture, nutrition and health sectors to address aflatoxins contamination better. Sustained public awareness at all levels, capacity building and aflatoxins related policies are necessary to support management initiatives

Aflatoxin contamination of non-cultivated fruits in Zambia

Open Access Journal; Published online: 09 Aug 2019Wild fruits are an important food and income source for many households in Zambia. Non-cultivated plants may be as susceptible as crops to aflatoxin contamination. Concentrations of aflatoxins in commonly consumed wild fruits from markets and characteristics of associated aflatoxin-producers need to be determined to assess the aflatoxin risk posed by handling, processing, storage, and consumption. Samples of Schinziophyton rautanenii (n = 22), Vangueriopsis lanciflora (n = 7), Thespesia garckeana (n = 17), Parinari curatellifolia (n = 17), Ziziphus spp. (n = 10), Adansonia digitata (n = 9), and Tamarindus indica (n = 23) were assayed for aflatoxin using lateral-flow immunochromatography from 2016 to 2017. Aflatoxins were above Zambia’s regulatory limit (10 μg/kg) in S. rautanenii (average = 57 μg/kg), V. lanciflora (average = 12 μg/kg), and T. garckeana (average = 11 μg/kg). The L strain morphotype of Aspergillus flavus was the most frequent member of Aspergillus section Flavi in market samples, although Aspergillus parasiticus and fungi with S morphology were also found. All fruits except T. indica supported both growth (mean = 3.1 × 108 CFU/g) and aflatoxin production (mean = 35,375 μg/kg) by aflatoxigenic Aspergillus section Flavi. Innate resistance to aflatoxin producers was displayed by T. indica. For the other fruits, environment and infecting fungi appeared to have the greatest potential to influence aflatoxin concentrations in markets. This is the first report of aflatoxins and aflatoxin-producers on native fruits in Zambia and suggests mitigation is required

Founder events influence structures of Aspergillus flavus populations

Open Access Article; Published online: 09 June 2020In warm regions, agricultural fields are occupied by complex Aspergillus flavus communities composed of isolates in many vegetative compatibility groups (VCGs) with varying abilities to produce highly toxic, carcinogenic aflatoxins. Aflatoxin contamination is reduced with biocontrol products that enable atoxigenic isolates from atoxigenic VCGs to dominate the population. Shifts in VCG frequencies similar to those caused by the introduction of biocontrol isolates were detected in Sonora, Mexico, where biocontrol is not currently practiced. The shifts were attributed to founder events. Although VCGs reproduce clonally, significant diversity exists within VCGs. Simple sequence repeat (SSR) fingerprinting revealed that increased frequencies of VCG YV150 involved a single haplotype. This is consistent with a founder event. Additionally, great diversity was detected among 82 YV150 isolates collected over 20 years across Mexico and the United States. Thirty‐six YV150 haplotypes were separated into two populations by Structure and SplitsTree analyses. Sixty‐five percent of isolates had MAT1‐1 and belonged to one population. The remaining had MAT1‐2 and belonged to the second population. SSR alleles varied within populations, but recombination between populations was not detected despite co‐occurrence at some locations. Results suggest that YV150 isolates with opposite mating‐type have either strongly restrained or lost sexual reproduction among themselves

Aflatoxin contamination of dried insects and fish in Zambia

Open Access Article; Published online: 17 Aug 2018Dried insects and fish are important sources of income and dietary protein in Zambia. Some aflatoxin-producing fungi are entomopathogenic and also colonize insects and fish after harvest and processing. Aflatoxins are carcinogenic, immune-suppressing mycotoxins that are frequent food contaminants worldwide. Several species within Aspergillus section Flavi have been implicated as causal agents of aflatoxin contamination of crops in Africa. However, aflatoxin producers associated with dried fish and edible insects in Zambia remain unknown, and aflatoxin concentrations in these foods have been inadequately evaluated. The current study sought to address these data gaps to assess potential human vulnerability through the dried fish and edible insect routes of aflatoxin exposure. Caterpillars (n = 97), termites (n = 4), and dried fish (n = 66) sampled in 2016 and 2017 were assayed for aflatoxin by using lateral flow immunochromatography. Average aflatoxin concentrations exceeded regulatory limits for Zambia (10 μg/kg) in the moth Gynanisa maja (11 μg/kg), the moth Gonimbrasia zambesina (Walker) (12 μg/kg), and the termite Macrotermes falciger (Gerstacker) (24 μg/kg). When samples were subjected to simulated poor storage, aflatoxins increased (P < 0.001) to unsafe levels in caterpillars (mean, 4,800 μg/kg) and fish (Oreochromis) (mean, 23 μg/kg). The L strain morphotype of A. flavus was the most common aflatoxin producer on dried fish (88% of Aspergillus section Flavi), termites (68%), and caterpillars (61%), with the exception of Gynanisa maja, for which A. parasiticus was the most common (44%). Dried fish and insects supported growth (mean, 1.3 × 109 CFU/g) and aflatoxin production (mean, 63,620 μg/kg) by previously characterized toxigenic Aspergillus section Flavi species, although the extent of growth and aflatoxigenicity depended on specific fungus-host combinations. The current study shows the need for proper storage and testing of dried insects and fish before consumption as measures to mitigate human exposure to aflatoxins through consumption in Zambia

Biological control of aflatoxins in maize and groundnut through use of aflasafe products developed for Ghana

United States Agency for International Developmen