Search

Aflatoxin contamination of important food and feed crops occurs frequently in warm tropical and subtropical regions. The contamination is caused mainly by Aspergillus flavus and A. parasiticus. Aflatoxin contamination negatively affects health and trade sectors and causes economic losses to agricultural industries. Many pre- and post-harvest technologies can limit aflatoxin contamination but may not always reduce aflatoxin concentrations below tolerance thresholds. However, the use of atoxigenic (non-toxin producing) isolates of A. flavus to competitively displace aflatoxin producers is a practical strategy that effectively limits aflatoxin contamination in crops from field to plate. Biocontrol products formulated with atoxigenic isolates as active ingredients have been registered for use in the US, several African nations, and one such product is in final stages of registration in Italy. Many other nations are seeking to develop biocontrol products to protect their crops. In this review article we present an overview of the biocontrol technology, explain the basis to select atoxigenic isolates as active ingredients, describe how formulations are developed and tested, and describe how a biocontrol product is used commercially. Future perspectives on formulations of aflatoxin biocontrol products, along with other important topics related to the aflatoxin biocontrol technology are also discussed. Fil: Moral, Juan. Universidad de Córdoba; España Fil: Garcia-Lopez, Maria Teresa. Universidad de Córdoba; España Fil: Camiletti, Boris Xavier. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto Multidisciplinario de Biología Vegetal. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas Físicas y Naturales. Instituto Multidisciplinario de Biología Vegetal; Argentina. Universidad Nacional de Córdoba. Facultad de Ciencias Agropecuarias. Departamento de Recursos Naturales. Cátedra de Microbiología Agrícola; Argentina Fil: Jaime, Ramon. University of California at Davis; Estados Unidos Fil: Michailides, Themis J.. University of California at Davis; Estados Unidos Fil: Bandyopadhyay, Ranajit. International Institute of Tropical Agriculture; Nigeria Fil: Ortega-Beltran, Alejandro. International Institute of Tropical Agriculture; Nigeria

Aflatoxins are secondary metabolites produced by soilborne saprophytic fungus Aspergillus flavus and closely related species that infect several agricultural commodities including groundnut and maize. The consumption of contaminated commodities adversely affects the health of humans and livestock. Aflatoxin contamination also causes significant economic and financial losses to producers. Research efforts and significant progress have been made in the past three decades to understand the genetic behavior, molecular mechanisms, as well as the detailed biology of host-pathogen interactions. A range of omics approaches have facilitated better understanding of the resistance mechanisms and identified pathways involved during host-pathogen interactions. Most of such studies were however undertaken in groundnut and maize. Current efforts are geared toward harnessing knowledge on host-pathogen interactions and crop resistant factors that control aflatoxin contamination. This study provides a summary of the recent progress made in enhancing the understanding of the functional biology and molecular mechanisms associated with host-pathogen interactions during aflatoxin contamination in groundnut and maize.

Aflatoxin contamination of groundnut and maize infected by Aspergillus section Flavi fungi is common throughout Senegal. The use of biocontrol products containing atoxigenic Aspergillus flavus strains to reduce crop aflatoxin content has been successful in several regions, but no such products are available in Senegal. The biocontrol product Aflasafe SN01 was developed for use in Senegal. The four active ingredients of Aflasafe SN01 are atoxigenic A. flavus genotypes native to Senegal and distinct from active ingredients used in other biocontrol products. Efficacy tests on groundnut and maize in farmers’ fields were carried out in Senegal during the course of 5 years. Active ingredients were monitored with vegetative compatibility analyses. Significant (P < 0.05) displacement of aflatoxin producers occurred in all years, districts, and crops. In addition, crops from Aflasafe SN01-treated fields contained significantly (P < 0.05) fewer aflatoxins both at harvest and after storage. Most crops from treated fields contained aflatoxin concentrations permissible in both local and international markets. Results suggest that Aflasafe SN01 is an effective tool for aflatoxin mitigation in groundnut and maize. Large-scale use of Aflasafe SN01 should provide health, trade, and economic benefits for Senegal. [Formula: see text] Copyright © 2020 The Author(s). This is an open access article distributed under the CC BY 4.0 International license .

Aflatoxin contamination in maize and groundnut is perennial in Ghana with substantial health and economic burden on the population. The present study examined for the first time the prevalence of aflatoxin contamination in maize and groundnut in major producing regions across three agroecological zones (AEZs) in Ghana. Furthermore, the distribution and aflatoxin-producing potential of Aspergillus species associated with both crops were studied. Out of 509 samples (326 of maize and 183 of groundnut), 35% had detectable levels of aflatoxins. Over 15% of maize and 11% of groundnut samples exceeded the aflatoxin threshold limits set by the Ghana Standards Authority of 15 and 20 ppb, respectively. Mycoflora analyses revealed various species and morphotypes within the Aspergillus section Flavi. A total of 5,083 isolates were recovered from both crops. The L morphotype of Aspergillus flavus dominated communities with 93.3% of the population, followed by Aspergillus spp. with S morphotype (6%), A. tamarii (0.4%), and A. parasiticus (0.3%). Within the L morphotype, the proportion of toxigenic members was significantly (P < 0.05) higher than that of atoxigenic members across AEZs. Observed and potential aflatoxin concentrations indicate that on-field aflatoxin management strategies need to be implemented throughout Ghana. The recovered atoxigenic L morphotype fungi are genetic resources that can be employed as biocontrol agents to limit aflatoxin contamination of maize and groundnut in Ghana. [Formula: see text] Copyright © 2018 The Author(s). This is an open access article distributed under the CC BY 4.0 International license .

Aspergillus flavus frequently contaminates maize, a critical staple for billions of people, with aflatoxins. Diversity among A. flavus L morphotype populations associated with maize in Sonora, Mexico was assessed and, in total, 869 isolates from 83 fields were placed into 136 vegetative compatibility groups (VCGs) using nitrate-nonutilizing mutants. VCG diversity indices did not differ in four agroecosystems (AES) but diversity significantly differed among years. Frequencies of certain VCGs changed manyfold over single years in both multiple fields and multiple AES. Certain VCGs were highly frequent (>1%) in 2006 but frequencies declined repeatedly in each of the two subsequent years. Other VCGs that had low frequencies in 2006 increased in 2007 and subsequently declined. None of the VCGs were consistently associated with any AES. Fourteen VCGs were considered dominant in at least a single year. However, frequencies often varied significantly among years. Only 9% of VCGs were detected all 3 years whereas 66% were detected in only 1 year. Results suggest that the most realistic measurements of both genetic diversity and the frequency of A. flavus VCGs are obtained by sampling multiple locations in multiple years. Single-season sampling in many locations should not be substituted for sampling over multiple years.

In 2015, all United Nations Member States adopted the 2030 Agenda for Sustainable Development to achieve peace and prosperity for all people in the planet. Meeting that ambitious agenda depends on fulfilling all objectives of 17 Sustainable Development Goals (SDGs). Multiple approaches by diverse actors, many of them interconnected, will allow achieving each SDG. However, with compromised food security and food safety, many SDGs will not be realized. In sub-Saharan Africa (SSA), maize and groundnut are two staple crops frequently contaminated with aflatoxins, which threaten food security and food safety. Aflatoxins are extremely dangerous compounds produced primarily by the fungus Aspergillus flavus. Even at minute concentrations, aflatoxins negatively influence health, income, and trade sectors. Farmers, traders, industries, and consumers become affected. However, practical solutions exist. Non-aflatoxin producing isolates (referred to as atoxigenic) of A. flavus can decrease crop aflatoxin content when used in biocontrol formulations to competitively displace aflatoxin producers during crop development. Typically, treated crops contain 80%–100% less aflatoxin than non-treated crops. The technology was developed by USDA-ARS for use in the US and has been adapted and improved for use in SSA where several products under the tradename Aflasafe are available. There are biocontrol products registered for use in 10 SSA countries and more are being developed. On the other hand, although highly effective, biocontrol is not a panacea. Less aflatoxin occurs across value chains when biocontrol is combined with other practices. In this review, we discuss how i) aflatoxin biocontrol products are developed, manufactured, licensed, and commercialized, ii) aflatoxin management strategies are designed, and iii) integrated aflatoxin management is or will soon be contributing to achieve, in several countries, many targets of most SDGs. We present integrated aflatoxin management as a model intervention contributing to tackle several challenges impeding prosperity and peace in SSA.

Aflatoxins pose significant food security and public health risks, decrease productivity and profitability of animal industries, and hamper trade. To minimize aflatoxin contamination in several crops, a biocontrol technology based on atoxigenic strains of Aspergillus flavus is commercially used in the United States and some African countries. Significant efforts are underway to popularize the use of biocontrol in Africa by various means including incentives. The purpose of this study was to develop quantitative pyrosequencing assays for rapid, simultaneous quantification of proportions of four A. flavus biocontrol genotypes within complex populations of A. flavus associated with maize crops in Nigeria to facilitate payment of farmer incentives for Aflasafe (a biocontrol product) use. Protocols were developed to confirm use of Aflasafe by small scale farmers in Nigeria. Nested PCR amplifications followed by sequence by synthesis pyrosequencing assays were required to quantify frequencies of the active ingredients and, in so doing, confirm successful use of biocontrol by participating farmers. The entire verification process could be completed in 3–4 days proving a savings over other monitoring methods in both time and costs and providing data in a time frame that could work with the commercial agriculture scheme. Quantitative pyrosequencing assays represent a reliable tool for rapid detection, quantification, and monitoring of multiple A. flavus genotypes within complex fungal communities, satisfying the requirements of the regulatory community and crop end-users that wish to determine which purchased crops were treated with the biocontrol product. Techniques developed in the current study can be modified for monitoring other crop-associated fungi.

In sub-Saharan Africa (SSA), diverse fungi belonging to Aspergillus section Flavi frequently contaminate staple crops with aflatoxins. Aflatoxins negatively impact health, income, trade, food security, and development sectors. Aspergillus flavus is the most common causal agent of contamination. However, certain A. flavus genotypes do not produce aflatoxins (i.e., are atoxigenic). An aflatoxin biocontrol technology employing atoxigenic genotypes to limit crop contamination was developed in the United States. The technology was adapted and improved for use in maize and groundnut in SSA under the trademark Aflasafe. Nigeria was the first African nation for which an aflatoxin biocontrol product was developed. The current study includes tests to assess biocontrol performance across Nigeria over the past decade. The presented data on efficacy spans years in which a relatively small number of maize and groundnut fields (8–51 per year) were treated through use on circa 36,000 ha in commercially-produced maize in 2018. During the testing phase (2009–2012), fields treated during one year were not treated in the other years while during commercial usage (2013–2019), many fields were treated in multiple years. This is the first report of a large-scale, long-term efficacy study of any biocontrol product developed to date for a field crop. Most (>95%) of 213,406 tons of maize grains harvested from treated fields contained 90%) contained 80% less aflatoxin content than untreated crops. The frequency of the biocontrol active ingredient atoxigenic genotypes in grains from treated fields was significantly higher than in grains from control fields. A higher proportion of grains from treated fields met various aflatoxin standards compared to grains from untreated fields. Results indicate that efficacy of the biocontrol product in limiting aflatoxin contamination is stable regardless of environment and cropping system. In summary, the biocontrol technology allows farmers across Nigeria to produce safer crops for consumption and increases potential for access to premium markets that require aflatoxin-compliant crops.

Coniothyrium glycines, the causal agent of red leaf blotch in soybeans, is considered a high-consequence biological agent. With limited genomic information known, there are no molecular genotyping or detection methods available. We report the draft genome sequences of three C. glycines isolates, greatly enhancing our knowledge of this species.

Las aflatoxinas son micotoxinas producidas por hongos de suelo del género Aspergillus, principalmente A. flavus y A. parasiticus. Por su alta toxicidad, están reguladas en numerosos alimentos para consumo humano y animal. Ambas especies crecen sobre restos de material vegetal produciendo un gran número de conidios aerovagantes que pueden colonizar y contaminar diferentes cultivos como maíz, cacahuete, algodón y frutos secos, entre otros. La contaminación por aflatoxinas en almendro y pistachero, sus causas y control, han sido intensamente estudiados en California en los últimos 25 años. En España, la situación es menos conocida y está siendo abordada en el marco de dos proyectos de investigación entre la Universidad de Córdoba y la Universidad de California, Davis. Aquí, revisamos la importancia actual, la etiología, biología, y control de esta problemática en los frutos secos españoles.

In California, aflatoxin contamination of almond, fig, and pistachio has become a serious problem in recent years due to long periods of drought and probably other climatic changes. The atoxigenic biocontrol product Aspergillus flavus AF36 has been registered for use to limit aflatoxin contamination of pistachio since 2012 and for use in almond and fig since 2017. New biocontrol technologies employ multiple atoxigenic genotypes because those provide greater benefits than using a single genotype. Almond, fig, and pistachio industries would benefit from a multi-strain biocontrol technology for use. Several A. flavus vegetative compatibility groups (VCGs) associated with almond, fig, and pistachio composed exclusively of atoxigenic isolates, including the VCG to which AF36 belongs to, YV36, were previously characterized in California. Here we report additional VCGs associated with two or the three crops. Representative isolates of 12 atoxigenic VCGs significantly (P < 0.001) reduced ( > 80%) aflatoxin accumulation in almond and pistachio when challenged with highly toxigenic isolates of A. flavus and A. parasiticus. Isolates of the evaluated VCGs, including AF36, constitute valuable endemic, well adapted, and efficient germplasm to design a multi-crop, multi-strain biocontrol strategy for use in tree crops in California. Availability of such strategy would favor long-term atoxigenic A. flavus communities across the affected areas of California and this would result in securing domestic and export markets for the nut crop and fig farmer industries and most importantly health benefits to consumers.

In warm agricultural areas across the globe, maize, groundnut, and other crops become frequently contaminated with aflatoxins produced primarily by the fungus Aspergillus flavus. Crop contamination with those highly toxic and carcinogenic compounds impacts both human and animal health, as well as the income of farmers and trade. In Nigeria, poultry productivity is hindered by high prevalence of aflatoxins in feeds. A practical solution to decrease crop aflatoxin content is to use aflatoxin biocontrol products based on non-toxin-producing strains of A. flavus. The biocontrol product Aflasafe&reg, was registered in 2014 for use in maize and groundnut grown in Nigeria. Its use allows the production of aflatoxin-safe maize and groundnut. A portion of the maize treated with Aflasafe in Nigeria is being used to manufacture feeds used by the poultry industry, and productivity is improving. One of the conditions to register Aflasafe with the national regulator was to demonstrate both the safety of Aflasafe-treated maize to avian species and the impact of Aflasafe as a public good. Results presented here demonstrate that the use of maize colonized by an atoxigenic strain of Aflasafe resulted in superior (p &lt, 0.05) broiler performance in all evaluated parameters in comparison to broilers fed with toxigenic maize. Use of an aflatoxin-sequestering agent (ASA) was not sufficient to counteract the harmful effects of aflatoxins. Both the safety and public good value of Aflasafe were demonstrated during our study. In Nigeria, the availability of aflatoxin-safe crops as a result of using Aflasafe allows poultry producers to improve their productivity, their income, and the health of consumers of poultry products.

Summary In 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.

Aflatoxins pose significant food security and public health risks, decrease productivity and profitability of animal industries, and hamper trade. To minimize aflatoxin contamination in several crops, a biocontrol technology based on atoxigenic strains of Aspergillus flavus is commercially used in the United States and some African countries. Significant efforts are underway to popularize the use of biocontrol in Africa by various means including incentives. The purpose of this study was to develop quantitative pyrosequencing assays for rapid, simultaneous quantification of proportions of four A. flavus biocontrol genotypes within complex populations of A. flavus associated with maize crops in Nigeria to facilitate payment of farmer incentives for Aflasafe (a biocontrol product) use. Protocols were developed to confirm use of Aflasafe by small scale farmers in Nigeria. Nested PCR amplifications followed by sequence by synthesis pyrosequencing assays were required to quantify frequencies of the active ingredients and, in so doing, confirm successful use of biocontrol by participating farmers. The entire verification process could be completed in 3–4 days proving a savings over other monitoring methods in both time and costs and providing data in a time frame that could work with the commercial agriculture scheme. Quantitative pyrosequencing assays represent a reliable tool for rapid detection, quantification, and monitoring of multiple A. flavus genotypes within complex fungal communities, satisfying the requirements of the regulatory community and crop end-users that wish to determine which purchased crops were treated with the biocontrol product. Techniques developed in the current study can be modified for monitoring other crop-associated fungi.

Across sub-Saharan Africa, chili peppers are fundamental ingredients of many traditional dishes. However, chili peppers may contain unsafe aflatoxin concentrations produced by Aspergillus section Flavi fungi. Aflatoxin levels were determined in chili peppers from three states in Nigeria. A total of 70 samples were collected from farmers&rsquo, stores and local markets. Over 25% of the samples contained unsafe aflatoxin concentrations. The chili peppers were associated with both aflatoxin producers and atoxigenic Aspergillus flavus genotypes. Efficacy of an atoxigenic biocontrol product, Aflasafe, registered in Nigeria for use on maize and groundnut, was tested for chili peppers grown in three states. Chili peppers treated with Aflasafe accumulated significantly less aflatoxins than nontreated chili peppers. The results suggest that Aflasafe is a valuable tool for the production of safe chili peppers. Use of Aflasafe in chili peppers could reduce human exposure to aflatoxins and increase chances to commercialize chili peppers in premium local and international markets. This is the first report of the efficacy of any atoxigenic biocontrol product for controlling aflatoxin in a spice crop.

Cowpea is an important protein source for human populations in many nations across sub-Saharan Africa (SSA). However, cowpea production is constrained by bacterial blight (CoBB) caused by Xanthomonas axonopodis pv. vignicola (Xav), a disease affecting most cowpea-growing areas. A large proportion of smallholder farmers across SSA rely on traditional cowpea landraces (CLR) to produce the crop. The International Institute of Tropical Agriculture (IITA) possesses the largest collection of cowpea germplasm, including several CLR accessions. However, screening for resistance to CoBB in most of the CLR accessions maintained at IITA has not been conducted. CoBB severity was evaluated in 103 CLR accessions from five African countries, the US, The Philippines, and Sri Lanka by artificially inoculating a highly virulent Xav strain in plants grown in a screenhouse. Highly significant (P

In California, aflatoxin contamination of almond, fig, and pistachio has become a serious problem in recent years due to long periods of drought and probably other climatic changes. The atoxigenic biocontrol product

Maize is an important staple crop for the majority of the population in Uganda. However, in tropical and subtropical climates, maize is frequently contaminated with aflatoxins, a group of cancer-causing and immuno-suppressive mycotoxins produced by Aspergillus section Flavi fungi. In Uganda, there is limited knowledge about the causal agents of aflatoxin contamination. The current study determined both the aflatoxin levels in pre-harvest maize across Uganda and the structures of communities of aflatoxin-producing fungi associated with the maize. A total of 256 pre-harvest maize samples were collected from 23 major maize-growing districts in eight agro-ecological zones (AEZ). Maize aflatoxin content ranged from 0 to 3760 ng/g although only around 5% for Ugandan thresholds. For EU it is about 16% of the samples contained aflatoxin concentrations above tolerance thresholds. A total of 3105 Aspergillus section Flavi isolates were recovered and these were dominated by the A. flavus L morphotype (89.4%). Densities of aflatoxin-producing fungi were negatively correlated with elevation. Farming systems and climatic conditions of the AEZ are thought to have influenced communities' structure composition. Fungi from different AEZ varied significantly in aflatoxin-producing abilities and several atoxigenic genotypes were identified. The extremely high aflatoxin concentrations detected in some of the studied regions indicate that management strategies should be urgently designed for use at the pre-harvest stage. Atoxigenic genotypes detected across Uganda could serve as aflatoxin biocontrol agents to reduce crop contamination from fields conditions and throughout the maize value chain.

Aflatoxin accumulation in maize (Zea mays L.) kernels is a serious economic and health problem that reduces grain quality and nutritional values and causes death to livestock and humans. Understanding the genetic parameters and heterotic responses of exotic maize inbred lines can facilitate their use for developing aflatoxin resistant parents of hybrids in Africa. This study was designed to (1) determine the heterotic affinities of aflatoxin resistant exotic lines, (2) identify exotic inbreds with good combining ability, and (3) determine the mode of inheritance of resistance to aflatoxin contamination in these lines. A line × tester mating design was used to determine combining ability of 12 yellow and 13 white inbreds and classify them into heterotic groups. The inbreds were crossed to two adapted testers representing two African heterotic groups and the resulting testcrosses along with hybrid checks were evaluated in separate trials at two locations for 2 years in Nigeria. General combining ability (GCA) effects were more important than specific combining ability effects for aflatoxin and grain yield. Among 15 exotic inbred lines having negative GCA effects for aflatoxin and 13 with positive GCA effects for grain yield, six combined the two desired traits. Five white and six yellow endosperm testcrosses were found to be good specific combiners for the two desired traits. The exotic lines with negative GCA effects for aflatoxin accumulation will be used as donor parents to develop backcross populations for generating new inbred lines with much higher levels of resistance to aflatoxin accumulation.