Aflatoxin Biocontrol Research Articles

Aflatoxin awareness and preventive agricultural practices are key to adoption of biocontrol among maize smallholder farmers in Tanzania.

Aflatoxins are toxic secondary metabolites produced by Aspergillus species that infect staple foods like maize causing threat to public health and economic impacts. The use of atoxigenic Aspergillus species is considered one of the promising technologies to prevent aflatoxin contamination in maize. Tanzania approved the use of aflatoxin biocontrol (Aflasafe®) in 2018 and introduced it to eight districts. Adoption and effectiveness of this technology depend on many factors including application of pre- and post-harvest practices. There is scant information on awareness of biocontrol and factors which influence the adoption and effectiveness of this technology. A cross-sectional study was conducted in Tanzania to assess awareness and identify factors influencing adoption of the technology. Data was collected from 334 smallholder farmers in Kiteto and Chemba districts and analyzed using SPSS version 20; p-values < 0.05 using a two-tailed test were considered statistically significant. Results indicated 95.4% are not aware and that only 2.7% of the farmers had used biocontrol technology. The use of biocontrol was significantly associated with high income level (p = 0.001) and exposure to print media (p = 0.03) and radio (p = 0.008). The use of hybrid seed (p = 0.01), grazing (p = 0.017), and rotation of crops on yearly basis (p = 0.024) were also significantly associated with the use of biocontrol. Income limits the use of biocontrol, requiring government subsidies for Aflasafe and a premium market for aflatoxin-free maize. Aflatoxin awareness and sensitization on adherence to good pre-harvest practices should be emphasized to enhance adoption of the technology.

Mitigation of aflatoxin contamination of maize, groundnut, and sorghum by commercial biocontrol products in farmers’ fields across Burkina Faso, Mali, Niger, and Togo

BackgroundAflatoxin contamination by Aspergillus section Flavi fungi poses a significant threat to food security and public health in sub-Saharan Africa (SSA). Maize, groundnut, and sorghum are staple crops frequently contaminated with aflatoxins, sometimes at dangerous levels. Despite its detrimental effects, many farmers in SSA lack access to effective tools for mitigating aflatoxin contamination. Biocontrol based on atoxigenic isolates of A. flavus is an effective tool to limit aflatoxin contamination.MethodsThe development, testing, registration, and commercial use of the aflatoxin biocontrol product Aflasafe BF01 for use in Burkina Faso is described. In addition, the deployment of the biocontrol technology across Mali, Niger, and Togo is documented, and for the first time, the use of aflatoxin biocontrol in sorghum is reported.ResultsIn all four countries, treated crops had significantly (P < 0.05) less aflatoxins than crops from untreated fields. Most treated crops met the stringent tolerance threshold for human consumption, 4 ppb total aflatoxin. Using native atoxigenic isolates of A. flavus and employing a multi-disciplinary approach, aflatoxin biocontrol products have demonstrated significant success in reducing aflatoxin levels in treated crops compared to untreated ones.ConclusionsThis multi-year, multi-funded source study underscores the effectiveness of biocontrol strategies in mitigating aflatoxin contamination at scale, offering a regional approach for sustainable management in West Africa and potentially unlocking significant health and economic benefits for the region.

Impact of storage conditions on the shelf life of aflatoxin biocontrol products containing atoxigenic isolates of Aspergillus flavus as active ingredient applied in various countries in Africa

Aflatoxin contamination significantly threatens food safety and security, particularly in tropical and sub-tropical regions where staple crops such as maize, groundnut, and sorghum become frequently affected. This contamination is primarily caused by the fungus Aspergillus flavus. The contamination causes adverse health effects, reduced income, and trade restrictions. In response to this challenge, various technologies have been developed to mitigate the impacts of aflatoxin. Among these, biocontrol products containing atoxigenic isolates of A. flavus as the active ingredient can effectively reduce aflatoxin levels both at pre- and post-harvest. A notable example of such products is Aflasafe, which contains four atoxigenic isolates native to specific target regions. These products have undergone rigorous testing, have received regulatory approval, and are commercially available in multiple African countries. However, their manufacturing processes have evolved, and comprehensive shelf life studies for current formulations are lacking. Evaluations of the spore production ability of atoxigenic A. flavus isolates in Aflasafe products over 4 years, under various storage conditions, revealed a significant linear decrease in sporulation with storage months (P < 0.001; R2 = 0.203), with no significant differences observed between treatments. However, this marginal decline (P = 0.398) is unlikely to be sufficient to prevent the effectiveness in limiting aflatoxin. In addition, storing the products for 2 weeks at 54 °C did not affect (P > 0.05) the ability of the coated fungi to produce spores compared to when the products were stored at 24 °C. The findings contribute valuable insights for manufacturers and users of atoxigenic-based aflatoxin biocontrol products, informing best practices for product storage and utilization to ensure prolonged effectivenes in aflatoxin mitigation efforts.

Structure of Aspergillus flavus populations associated with maize in Greece, Spain, and Serbia: Implications for aflatoxin biocontrol on a regional scale.

Aspergillus flavus is the most frequently identified producer of aflatoxins. Non-aflatoxigenic members of the A. flavus L strains are used in various continents as active ingredients of bioprotectants directed at preventing aflatoxin contamination by competitive displacement of aflatoxin producers. The current research examined the genetic diversity of A. flavus L strain across southern Europe to gain insights into the population structure and evolution of this species and to evaluate the prevalence of genotypes closely related to MUCL54911, the active ingredient of AF-X1. A total of 2173L strain isolates recovered from maize collected across Greece, Spain, and Serbia in 2020 and 2021 were subjected to simple sequence repeat (SSR) genotyping. The analysis revealed high diversity within and among countries and dozens of haplotypes shared. Linkage disequilibrium analysis indicated asexual reproduction and clonal evolution of A. flavus L strain resident in Europe. Moreover, haplotypes closely related to MUCL54911 were found to belong to the same vegetative compatibility group (VCG) IT006 and were relatively common in all three countries. The results indicate that IT006 is endemic to southern Europe and may be utilized as an aflatoxin mitigation tool for maize across the region without concern for potential adverse impacts associated with the introduction of an exotic microorganism.

Efficacy of atoxigenic Aspergillus flavus from southern China as biocontrol agents against aflatoxin contamination in corn and peanuts

Aspergillus flavus is a ubiquitous facultative pathogen that routinely infects important crops leading to formation of aflatoxins during crop development and after harvest. Corn and peanuts in warm and/or drought-prone regions are highly susceptible to aflatoxin contamination. Controlling aflatoxin using atoxigenic A. flavus is a widely adopted strategy. However, no A. flavus genotypes are currently approved for use in China. The current study aimed to select atoxigenic A. flavus endemic to Guangxi Zhuang Autonomous Region with potential as active ingredients of aflatoxin biocontrol products. A total of 204 A. flavus isolates from corn, peanuts, and field soil were evaluated for ability to produce the targeted mycotoxins. Overall, 57.3% could not produce aflatoxins while 17.15% were incapable of producing both aflatoxins and CPA. Atoxigenic germplasm endemic to Guangxi was highly diverse, yielding 8 different gene deletion patterns in the aflatoxin and CPA biosynthesis gene clusters ranging from no deletion to deletion of both clusters. Inoculation of corn and peanuts with both an aflatoxin producer and selected atoxigenic genotypes showed significant reduction (74 to 99%) in aflatoxin B1 (AFB1) formation compared with inoculation with the aflatoxin producer alone. Atoxigenic genotypes also efficiently degraded AFB1 (61%). Furthermore, atoxigenic isolates were also highly efficient at reducing aflatoxin concentrations even when present at lower concentrations than aflatoxin producers. The use of multiple atoxigenics was not always as effective as the use of a single atoxigenic. Effective atoxigenic genotypes of A. flavus with known mechanisms of atoxigenicity are demonstrated to be endemic to Southern China. These A. flavus may be utilized as active ingredients of biocontrol products without concern for detrimental impacts that may result from introduction of exotic fungi. Field efficacy trials in the agroecosystems of Southern China are needed to determine the extent to which such products may allow the production of safer food and feed.

Telomere-to-telomere genome assembly of the aflatoxin biocontrol agent Aspergillus flavus isolate La3279 isolated from maize in Nigeria.

Here, we report the complete genome of the non-aflatoxigenic Aspergillus flavus isolate La3279, which is an active ingredient of the aflatoxin biocontrol product Aflasafe. The chromosome-scale assembly clarifies the deletion pattern in the aflatoxin biosynthesis gene cluster and corrects a misidentified assembly previously published for this isolate.

Genetic Diversity of Aspergillus flavus on Maize in Guatemala.

Aflatoxin contamination of maize is a leading threat to health in Guatemala. This contamination is the result of infection from Aspergillus flavus and has been effectively reduced in other countries through application of nonaflatoxigenic, indigenous strains of A. flavus. We collected 82 maize samples from throughout Guatemala in two years and isolated 272 A. flavus from these samples, including 126 unique genotypes. We provide here a phenotypic and simple sequence repeat (SSR)-based genotypic description of these isolates, as well as an analysis of the diversity of this population. High levels of genetic diversity were observed with the nonaflatoxigenic isolates in this study, but this information contributes to the development of indigenous aflatoxin biocontrol products.

Approach for quick exploration of highly effective broad-spectrum biocontrol strains based on PO8 protein inhibition

Aflatoxin is a group of strongly toxic and carcinogenic mycotoxins produced by Aspergillus flavus and other Aspergillus species, which caused food contamination and food loss problems widely across the world especially in developing countries, thus threatening human health and sustainable development. So, it is important to develop new, green, and broad-spectrum biocontrol technology for the prevention of aflatoxin contamination sources. Previously, we found that the PO8 protein from aflatoxigenic A. flavus could be used as a biomarker to predict aflatoxin production in peanuts (so the PO8 is named as an early warning molecule), which infers that the PO8 is relative to aflatoxin production. Therefore, in the study, based on inhibiting the PO8, a new and quick strategy for screening aflatoxin biocontrol strains for developing control agents was presented. With the PO8 inhibition method, four biocontrol strains (2 strains were isolated from peanut kernels with sterilized surface and another 2 strains from peanut rhizosphere soil) were selected and combined to increase prevention wide-spectrum. As a result, the combination showed over 90% inhibition to all tested aflatoxigenic A. flavus isolated from three different peanut production areas (north, middle, and south areas of China), and better than any single strain. The field experiments located in five provinces of China showed that the practice prevention effects (inhibition of aflatoxigenic fungi on the surface of the peanuts) were from 50% to over 80%. The results indicated that the strategy of inhibiting the early warning molecule PO8 can be used to develop aflatoxin control agents well.

Multiple Year Influences of the Aflatoxin Biocontrol Product AF-X1 on the A. flavus Communities Associated with Maize Production in Italy.

AF-X1 is a commercial aflatoxin biocontrol product containing the non-aflatoxigenic (AF-) strain of Aspergillus flavus MUCL54911 (VCG IT006), endemic to Italy, as an active ingredient. The present study aimed to evaluate the long-term persistence of VCG IT006 in the treated fields, and the multi-year influence of the biocontrol application on the A. flavus population. Soil samples were collected in 2020 and 2021 from 28 fields located in four provinces in north Italy. A vegetative compatibility analysis was conducted to monitor the occurrence of VCG IT006 on the total of the 399 isolates of A. flavus that were collected. IT006 was present in all the fields, mainly in the fields treated for 1 yr or 2 consecutive yrs (58% and 63%, respectively). The densities of the toxigenic isolates, detected using the aflR gene, were 45% vs. 22% in the untreated and treated fields, respectively. After displacement via the AF- deployment, a variability from 7% to 32% was noticed in the toxigenic isolates. The current findings support the long-term durability of the biocontrol application benefits without deleterious effects on each fungal population. Nevertheless, based on the current results, as well as on previous studies, the yearly applications of AF-X1 to Italian commercial maize fields should continue.

Aflatoxin biocontrol in practice requires a multidisciplinary, long-term approach

One of the most elusive food safety problems is the contamination of staple crops with the highly carcinogenic aflatoxins produced byAspergillussection Flavi fungi. Governments, farmers, institutions, consumers, and companies demand aflatoxin solutions. Many aflatoxin management technologies exist, but their real-life use and effectiveness is determined by diverse factors. Biocontrol products based on atoxigenic isolates ofA. flavuscan effectively reduce aflatoxins from field to fork. However, development, testing, and registration of this technology is a laborious process. Further, several barriers prevent the sustainable use of biocontrol products. There are challenges to have the products accepted, to make them available at scale and develop mechanisms for farmers to buy them, to have the products correctly used, to demonstrate their value, and to link farmers to buyers of aflatoxin-safe crops. Developing an effective aflatoxin management technology is the first, major step. The second one, perhaps more complicated and unfortunately seldomly discussed, is to develop mechanisms to have it used at scale, sustainably, and converged with other complementary technologies. Here, challenges and actions to scale the aflatoxin biocontrol technology in several countries in sub-Saharan Africa are described with a view to facilitating aflatoxin management efforts in Africa and beyond.

Identification of naturally occurring atoxigenic strains of Fusarium verticillioides and their potential as biocontrol agents of mycotoxins and ear rot pathogens of maize

Pre-harvest infection of seed with either of two major ear rot pathogens of maize, Fusarium verticillioides and Aspergillus flavus, leads to high economic losses and detrimental health effects to humans and animals through the production of mycotoxins, fumonisin and aflatoxin, respectively. These carcinogenic compounds are some of the most agriculturally important mycotoxins, consumption of which can lead to various cancers and stunted growth. While atoxigenic strains of A. flavus are used for biocontrol of aflatoxin, no similar biocontrol agent exists for F. verticillioides. We hypothesized that fumonisin non-producing (atoxigenic) F. verticillioides may competitively exclude or antagonize A. flavus and promote plant growth. We screened 150 isolates of Fusarium spp. collected from maize across Texas and identified several F. verticillioides isolates that did not produce fumonisin either in vitro or when grown on maize kernels. Atoxigenic strains of F. verticillioides, 302-A6 and PAL 401, variably affected plant growth following seed coating or soil injection. When pre-applied in a kernel bioassay, both strains significantly inhibited aflatoxin contamination compared to kernels later inoculated only with A. flavus. Applied as a seed coating, neither isolate inhibited aflatoxin or fumonisin accumulation in mature plants. Results confirmed that fumonisin is not required for virulence in Fusarium stalk rot infections. Application of F. verticillioides 302-A6 to developing ears reduced fumonisin accumulation by naturally-occurring toxigenic strains of F. verticillioides. This atoxigenic strain of F. verticillioides and others need to be further investigated to prove their efficacy and potential use as atoxigenic biocontrol agents to reduce fumonisin risk in corn.

Impact of frequency of application on the long-term efficacy of the biocontrol product Aflasafe in reducing aflatoxin contamination in maize.

Aflatoxins, produced by several Aspergillus section Flavi species in various crops, are a significant public health risk and a barrier to trade and development. In sub-Saharan Africa, maize and groundnut are particularly vulnerable to aflatoxin contamination. Aflasafe, a registered aflatoxin biocontrol product, utilizes atoxigenic A. flavus genotypes native to Nigeria to displace aflatoxin producers and mitigate aflatoxin contamination. Aflasafe was evaluated in farmers' fields for 3 years, under various regimens, to quantify carry-over of the biocontrol active ingredient genotypes. Nine maize fields were each treated either continuously for 3 years, the first two successive years, in year 1 and year 3, or once during the first year. For each treated field, a nearby untreated field was monitored. Aflatoxins were quantified in grain at harvest and after simulated poor storage. Biocontrol efficacy and frequencies of the active ingredient genotypes decreased in the absence of annual treatment. Maize treated consecutively for 2 or 3 years had significantly (p < 0.05) less aflatoxin (92% less) in grain at harvest than untreated maize. Maize grain from treated fields subjected to simulated poor storage had significantly less (p < 0.05) aflatoxin than grain from untreated fields, regardless of application regimen. Active ingredients occurred at higher frequencies in soil and grain from treated fields than from untreated fields. The incidence of active ingredients recovered in soil was significantly correlated (r = 0.898; p < 0.001) with the incidence of active ingredients in grain, which in turn was also significantly correlated (r = -0.621, p = 0.02) with aflatoxin concentration. Although there were carry-over effects, caution should be taken when drawing recommendations about discontinuing biocontrol use. Cost-benefit analyses of single season and carry-over influences are needed to optimize use by communities of smallholder farmers in sub-Saharan Africa.

Characterization of the Aspergillus flavus Population from Highly Aflatoxin-Contaminated Corn in the United States.

Aflatoxin contamination of corn is a major threat to the safe food and feed. The United States Federal Grain Inspection Service (FGIS) monitors commercial grain shipments for the presence of aflatoxin. A total of 146 Aspergillus flavus were isolated from 29 highly contaminated grain samples to characterize the visual phenotypes, aflatoxin-producing potential, and genotypes to explore the etiological cause of high aflatoxin contamination of US corn. Five of the isolates had reduced sensitivity (43-49% resistant) to the fungicide azoxystrobin, with the remainder all being over 50% resistant to azoxystrobin at the discriminating dose of 2.5 µg/mL. Only six isolates of the highly aflatoxigenic S morphotype were found, and 48 isolates were non-aflatoxigenic. Analysis of the mating type locus revealed 45% MAT 1-1 and 55% MAT 1-2. The A. flavus population originating from the highly aflatoxin contaminated grain samples was compared to a randomly selected subset of isolates originating from commercial corn samples with typical levels of aflatoxin contamination (average &lt; 50 ppb). Use of simple sequence repeat (SSR) genotyping followed by principal component analysis (PCoA) revealed a similar pattern of genotypic distribution in the two populations, but greater diversity in the FGIS-derived population. The noticeable difference between the two populations was that genotypes identical to strain NRRL 21882, the active component of the aflatoxin biocontrol product Afla-Guard™, were ten times more common in the commercial corn population of A. flavus compared to the population from the high-aflatoxin corn samples. The other similarities between the two populations suggest that high aflatoxin concentrations in corn grain are generally the result of infection with common A. flavus genotypes.

Aflatoxin biocontrol effectiveness in the real world-Private sector-led efforts to manage aflatoxins in Nigeria through biocontrol-centered strategies.

Aflatoxins are toxic compounds produced by several Aspergillus species that contaminate various crops. The impact of aflatoxin on the health of humans and livestock is a concern across the globe. Income, trade, and development sectors are affected as well. There are several technologies to prevent aflatoxin contamination but there are difficulties in having farmers use them. In Nigeria, an aflatoxin biocontrol product containing atoxigenic isolates of A. flavus has been registered with regulatory authorities and is now being produced at scale by the private company Harvestfield Industries Limited (HIL). The current study reports results of biocontrol effectiveness trials in maize conducted by HIL during 2020 in several locations across Nigeria and compared to untreated maize from nearby locations. Also, maize was collected from open markets to assess levels of contamination. All treated maize met tolerance thresholds (i.e., <4 ppb total aflatoxin). In contrast, most maize from untreated fields had a higher risk of aflatoxin contamination, with some areas averaging 38.5 ppb total aflatoxin. Maize from open markets had aflatoxin above tolerance thresholds with even an average of up to 90.3 ppb. Results from the trials were presented in a National Workshop attended by key officers of Government agencies, farmer organizations, the private sector, NGOs, and donors. Overall, we report (i) efforts spearheaded by the private sector to have aflatoxin management strategies used at scale in Nigeria, and (ii) deliberations of key stakeholders to ensure the safety of crops produced in Nigeria for the benefit of farmers, consumers, and industries.

Biocontrol of Aflatoxin through Biodegradation by Using Environment Friendly Microbes

Biocontrol of Aflatoxin through Biodegradation by Using Environment Friendly Microbes

Development of biochar-impregnated alginate beads for the delivery of biocontrol agents for peanut aflatoxin

The competitive inhibition of aflatoxigenic fungi by non-aflatoxigenic Aspergillus flavus has proved to be an effective method to prevent and control peanut aflatoxin contamination, and most of the currently used inoculum carriers are grains. In this study, the reliability and efficiency of replacing grain kernels with novel chitosan-coated alginate-poly(N-isopropylacrylamide) (PNIPAAm) beads impregnated with biochar (CSACB) were evaluated. Characterisation of the beads was performed by SEM, thermogravimetry analysis (TGA), and swelling properties analyses. The optimised CSACB beads had good physical stability, shelf life, and entrapment efficiency. In addition, the water-holding capacity and porous structure were excellent, as the biochar provided a beneficial microenvironment for the attachment and microbial growth of the biocontrol fungus. The effect of reducing aflatoxin in peanuts was verified experimentally. Collectively, the novel CSACB beads are suitable carriers of non-aflatoxigenic A. flavus for the biocontrol of peanut aflatoxin.

Genetic Diversity of Aspergillus flavus Associated with Chili in Nigeria and Identification of Haplotypes with Potential in Aflatoxin Mitigation.

Dried red chili (Capsicum spp.), a widely produced and consumed spice in Nigeria, is often contaminated by aflatoxins. Aflatoxins are potent mycotoxins of severe health and economic concern worldwide. Aspergillus flavus often contaminates crops with aflatoxins in warm regions; however, not all isolates are aflatoxin producers. Nonaflatoxigenic isolates have potential as biocontrol agents for aflatoxin mitigation. The current study examined the genetic diversity of A. flavus (n = 325) associated with chilies in Nigeria and identified 123 nonaflatoxigenic isolates. The Nigerian A. flavus isolates from chili were diverse at 17 microsatellite loci, with 5 to 36 alleles per locus, and included 152 haplotypes. The isolates that are active ingredients in Aflasafe, registered for aflatoxin biocontrol on maize and groundnuts in Nigeria, did not share haplotypes with the chili isolates. Of the 152 haplotypes, 65% produced aflatoxins in autoclaved maize, some of which (17%) produced >100,000 µg/kg of aflatoxins. Aflatoxins were not detected in 35% of the haplotypes. Cluster amplification pattern assay detected large deletions in the aflatoxin biosynthetic clusters of some (32%) of the nonaflatoxigenic haplotypes. Coinfection of chili with nonaflatoxigenic isolates from chilies (n = 7) and A. aflatoxiformans resulted in a significantly greater average reduction in total aflatoxins compared with that achieved by Aflasafe active ingredient isolates (P < 0.01). These nonaflatoxigenic isolates are a genetic resource for the development of biological control products for aflatoxin mitigation in chilies in Nigeria and should be evaluated under field conditions.

Resistance to Aspergillus flavus and Aspergillus parasiticus in Almond Advanced Selections and Cultivars and Its Interaction with the Aflatoxin Biocontrol Strategy.

Aflatoxin contamination of almond kernels, caused by Aspergillus flavus and A. parasiticus, is a severe concern for growers because of its high toxicity. In California, the global leader of almond production, aflatoxin can be managed by applying the biological control strain AF36 of A. flavus and selecting resistant cultivars. Here, we classified the almond genotypes by K-Means cluster analysis into three groups (susceptible [S], moderately susceptible [MS], or resistant [R]) based on aflatoxin content of inoculated kernels. The protective effects of the shell and seedcoat in preventing aflatoxin contamination were also examined. The presence of intact shells reduced aflatoxin contamination >100-fold. The seedcoat provided a layer of protection but not complete protection. In kernel inoculation assays, none of the studied almond genotypes showed a total resistance to the pathogen. However, nine traditional cultivars and four advanced selections were classified as R. Because these advanced selections contained germplasm derived from peach, we compared the kernel resistance of three peach cultivars to that shown by kernels of an R (Sonora) and an S (Carmel) almond cultivar and five pistachio cultivars. Overall, peach kernels were significantly more resistant to the pathogen than almond kernels, which were more resistant than pistachio kernels. Finally, we studied the combined effect of the cultivar resistance and the biocontrol strain AF36 in limiting aflatoxin contamination. For this, we coinoculated almond kernels of R Sonora and S Carmel with AF36 72 h before or 48 h after inoculating with an aflatoxin-producing strain of A. flavus. The percentage of aflatoxin reduction by AF36 strain was greater in kernels of Carmel (98%) than in those of Sonora (83%). Cultivar resistance also affected the kernel colonization by the biological control strain. AF36 strain limited aflatoxin contamination in almond kernels even when applied 48 h after the aflatoxin-producing strain. Our results show that biocontrol combined with the use of cultivars with resistance to aflatoxin contamination can result in a more robust protection strategy than the use of either practice in isolation.

Atoxigenic-based technology for biocontrol of aflatoxin in maize and groundnuts for Tanzania

Application of biocontrol products containing atoxigenic isolates ofAspergillus flavus to reduce aflatoxin content in crops is an effective strategy for managing aflatoxin in several regions throughout the world. We report the development and validation of two aflatoxin biocontrol products, Aflasafe TZ01 and Aflasafe TZ02, for use in maize and groundnut in Tanzania, a country frequently affected by aflatoxin contamination. Each product contains four atoxigenicA. flavus genotypes native and widely distributed in Tanzania. Efficacy tests on maize and groundnut were conducted over two years and in four regions of Tanzania where aflatoxin contamination is prevalent. Application of both products significantly (P&lt;0.05) reduced aflatoxin levels in maize and groundnut in both years and in all districts. No differences were observed in totalAspergillus sectionFlavi population in treated and untreated fields, revealing that application of the biocontrol products do not alter overallAspergillus populations in the environment. The results indicate that both products are effective tools for aflatoxin mitigation in groundnut and maize. The products were officially registered in 2018. Currently, there are scale-out and-up efforts of aflatoxin biocontrol products in Tanzania through a private sector company that is making the products available to farmers. Protecting maize and groundnut from aflatoxin contamination in Tanzania can result in health, income, and trade benefits.

Degradation of Aflatoxins B1 by Atoxigenic Aspergillus flavus Biocontrol Agents.

Aflatoxins are potent Aspergillus mycotoxins that contaminate food and feed, thereby impacting health and trade. Biopesticides with atoxigenic Aspergillus flavus isolates as active ingredients are used to reduce aflatoxin contamination in crops. The mechanism of aflatoxin biocontrol is primarily attributed to competitive exclusion but, sometimes, aflatoxin is reduced by greater amounts than can be explained by displacement of aflatoxin-producing fungi on the crop. Objectives of this study were to (i) evaluate the ability of atoxigenic A. flavus genotypes to degrade aflatoxin B1 (AFB1) and (ii) characterize impacts of temperature, time, and nutrient availability on AFB1 degradation by atoxigenic A. flavus. Aflatoxin-contaminated maize was inoculated with atoxigenic isolates in three separate experiments that included different atoxigenic genotypes, temperature, and time as variables. Atoxigenic genotypes varied in aflatoxin degradation but all degraded AFB1 >44% after 7 days at 30°C. The optimum temperature for AFB1 degradation was 25 to 30°C, which is similar to the optimum range for AFB1 production. In a time-course experiment, atoxigenics degraded 40% of AFB1 within 3 days, and 80% of aflatoxin was degraded by day 21. Atoxigenic isolates were able to degrade and utilize AFB1 as a sole carbon source in a chemically defined medium but quantities of AFB1 degraded declined as glucose concentrations increased. Degradation may be an additional mechanism through which atoxigenic A. flavus biocontrol products reduce aflatoxin contamination pre- or postharvest. Thus, selection of optimal atoxigenic active ingredients can include assessment of both competitive ability in agricultural fields and their ability to degrade aflatoxins.