Sexuality generates diversity in the aflatoxin gene cluster: evidence on a global scale.

Aflatoxins are produced by Aspergillus flavus and A. parasiticus in oil-rich seed and grain crops and are a serious problem in agriculture, with aflatoxin B1 being the most carcinogenic natural compound known. Sexual reproduction in these species occurs between individuals belonging to different vegetative compatibility groups (VCGs). We examined natural genetic variation in 758 isolates of A. flavus, A. parasiticus and A. minisclerotigenes sampled from single peanut fields in the United States (Georgia), Africa (Benin), Argentina (Córdoba), Australia (Queensland) and India (Karnataka). Analysis of DNA sequence variation across multiple intergenic regions in the aflatoxin gene clusters of A. flavus, A. parasiticus and A. minisclerotigenes revealed significant linkage disequilibrium (LD) organized into distinct blocks that are conserved across different localities, suggesting that genetic recombination is nonrandom and a global occurrence. To assess the contributions of asexual and sexual reproduction to fixation and maintenance of toxin chemotype diversity in populations from each locality/species, we tested the null hypothesis of an equal number of MAT1-1 and MAT1-2 mating-type individuals, which is indicative of a sexually recombining population. All samples were clone-corrected using multi-locus sequence typing which associates closely with VCG. For both A. flavus and A. parasiticus, when the proportions of MAT1-1 and MAT1-2 were significantly different, there was more extensive LD in the aflatoxin cluster and populations were fixed for specific toxin chemotype classes, either the non-aflatoxigenic class in A. flavus or the B1-dominant and G1-dominant classes in A. parasiticus. A mating type ratio close to 1∶1 in A. flavus, A. parasiticus and A. minisclerotigenes was associated with higher recombination rates in the aflatoxin cluster and less pronounced chemotype differences in populations. This work shows that the reproductive nature of the population (more sexual versus more asexual) is predictive of aflatoxin chemotype diversity in these agriculturally important fungi.

Mites associated with stored grain commodities in Benin, West Africa.

After insects, mites are the major arthropod pests that inhabit stored agricultural products worldwide. To determine the acarofauna that infests cowpea, maize, paddy rice and sorghum in Benin (West Africa), surveys were conducted in some principal markets (Dantokpa, Glazoue and Parakou) of this country. A total of 555 samples of grains and debris were collected in May and September 2011. More than 56 species belonging to 24 mite families were recorded in the four products. These mite species included predators, parasites, fungivorous, phytophagous and other groups whose feeding habits are not well known. The family Cheyletidae was the most prevalent and the most diverse predatory mite family encountered, in which Cheyletus malaccensis Oudemans was the most abundant species. Several families of mite pests and mites responsible for allergies (Acaridae, Glycyphagidae, Pyroglyphidae, Pyemotidae and Saproglyphidae) were also detected. The three most dominant and frequent species were C. malaccensis, Suidasia nesbitti (Hughes) and Suidasia sp. Statistical analysis showed that densities of these three mite species were higher in Parakou than in Glazoue and Dantokpa, on one hand, and higher in debris than in grains, on the other hand. The densities of S. nesbitti and Suidasia sp. decreased significantly during the dry season, whereas C. malaccensis remained stable throughout the two samplings. Of all grains, sorghum was the least infested with mites. This study shows that in Benin mites are present in stored agricultural products to which they cause serious damage, and may cause various allergies to people.

Potential use of the bushmint, Hyptis suaveolens, for the control of infestation by the pink stalk borer, Sesamia calamistis on maize in southern Benin, West Africa.

Maize production in Benin, especially in resource-poor farmers' fields, is constrained by stemborers among other factors. One of the major stemborers in southern Benin is Sesamia calamistis Hampson (Lepidoptera: Noctuidae). African farmers cannot afford to use commercial insecticides for controlling stemborers - they are expensive and unsuitable for durable pest management systems due to eco-toxicity. There is therefore a need for cheaper and environmentally friendly methods and botanicals offer an attractive alternative. The bushmint, Hyptis suaveolens (L.) Poit. (Lamiales: Lamiaceae), was compared with the commercial insecticide Furadan (carbofuran) for the control of S. calamistis on maize Zea mays L. (Poales: Poaceae). Trials were conducted in the screenhouse and in the field during the minor cropping season in 2004 at the International Institute of Tropical Agriculture (IITA)-Benin station. The variables measured included numbers of egg masses per plant, eggs per egg mass (in the screenhouse study), population density of S. calamistis, percentage of infested plants and/or ears, and deadhearts in the field. Irrespective of the variable considered, the aqueous extract of H. suaveolens compared favorably with Furadan while maize surrounded by live H. suaveolens plants had lower S. calamistis densities.

Effect of storage conditions on microbiological and physicochemical quality of shea butter.

Storage conditions are key constraints for quality assurance of the shea (Vitellaria paradoxa Gaertner) butter. In the Sudan savannah Africa, storage conditions of butter produced by women vary across and among processors, traders and consumers. These conditions could impact the quality of the products and reduced their access to international market. The present study attempted to investigate the effect of storage duration and packaging materials on microbiological and physicochemical characteristics of shea butter under tropical climatic conditions. Five packaging materials traditionally used in shea butter value chain were tested for their efficacy in storing shea butter freshly produced. Total germs, yeasts and mould varied with packaging materials and storage duration. After 2 months of storage, moisture content of butter remained constant (5%) whereas acid value increased from 3.3 to 5.4 mg KOH/g, peroxide value from 8.1 to 10.1 meq O2/kg and iodine value dropped from 48.8 to 46.2 mg I2/100 g in shea butter irrespectively to the storage materials used. The basket papered with jute bag was the less effective in ensuring the quality of butter during storage while plastic containers and plastic bags seemed to be the best packaging materials.

Workgroup report: public health strategies for reducing aflatoxin exposure in developing countries.

Consecutive outbreaks of acute aflatoxicosis in Kenya in 2004 and 2005 caused > 150 deaths. In response, the Centers for Disease Control and Prevention and the World Health Organization convened a workgroup of international experts and health officials in Geneva, Switzerland, in July 2005. After discussions concerning what is known about aflatoxins, the workgroup identified gaps in current knowledge about acute and chronic human health effects of aflatoxins, surveillance and food monitoring, analytic methods, and the efficacy of intervention strategies. The workgroup also identified public health strategies that could be integrated with current agricultural approaches to resolve gaps in current knowledge and ultimately reduce morbidity and mortality associated with the consumption of aflatoxin-contaminated food in the developing world. Four issues that warrant immediate attention were identified: a) quantify the human health impacts and the burden of disease due to aflatoxin exposure; b) compile an inventory, evaluate the efficacy, and disseminate results of ongoing intervention strategies; c) develop and augment the disease surveillance, food monitoring, laboratory, and public health response capacity of affected regions; and d) develop a response protocol that can be used in the event of an outbreak of acute aflatoxicosis. This report expands on the workgroup's discussions concerning aflatoxin in developing countries and summarizes the findings.

Diversity in Secondary Metabolites Including Mycotoxins from Strains of Aspergillus Section Nigri Isolated from Raw Cashew Nuts from Benin, West Africa.

In a previous study, raw cashew kernels were assayed for the fungal contamination focusing on strains belonging to the genus Aspergillus and on aflatoxins producers. These samples showed high contamination with Aspergillus section Nigri species and absence of aflatoxins. To investigate the diversity of secondary metabolites, including mycotoxins, the species of A. section Nigri may produce and thus threaten to contaminate the raw cashew kernels, 150 strains were isolated from cashew samples and assayed for their production of secondary metabolites using liquid chromatography high resolution mass spectrometry (LC-HRMS). Seven species of black Aspergilli were isolated based on morphological and chemical identification: A. tubingensis (44%), A. niger (32%), A. brasiliensis (10%), A. carbonarius (8.7%), A. luchuensis (2.7%), A. aculeatus (2%) and A. aculeatinus (0.7%). From these, 45 metabolites and their isomers were identified. Aurasperone and pyranonigrin A, produced by all species excluding A. aculeatus and A. aculeatinus, were most prevalent and were encountered in 146 (97.3%) and 145 (95.7%) isolates, respectively. Three mycotoxins groups were detected: fumonisins (B2 and B4) (2.7%) ochratoxin A (13.3%), and secalonic acids (2%), indicating that these mycotoxins could occur in raw cashew nuts. Thirty strains of black Aspergilli were randomly sampled for verification of species identity based on sequences of ß-tubulin and calmodulin genes. Among them, 27 isolates were positive to the primers used and 11 were identified as A. niger, 7 as A. tubingensis, 6 as A. carbonarius, 2 as A. luchuensis and 1 as A. welwitschiae confirming the species names as based on morphology and chemical features. These strains clustered in 5 clades in A. section Nigri. Chemical profile clustering also showed also 5 groups confirming the species specific metabolites production.

Effect of essential oils on the growth of Fusarium verticillioides and fumonisin contamination in corn.

Essential oils extracted by hydrodistillation from local plants in Benin, western Africa, and oil from seeds of the neem tree (Azadirachta indica) were evaluated in vitro and in vivo for their efficacy against Fusarium verticillioides infection and fumonisin contamination. Fumonisin in corn was quantified using a fluorometer and the Vicam method. Oils from Cymbopogon citratus, Ocimum basilicum, and Ocimum gratissimum were the most effective in vitro, completely inhibiting the growth of F. verticillioides at lower concentrations over 21 days of incubation. These oils reduced the incidence of F. verticillioides in corn and totally inhibited fungal growth at concentrations of 8, 6.4, and 4.8 microL/g, respectively, over 21 days. At the concentration of 4.8 microL/g, these oils did not affect significantly fumonisin production. However, a marked reduction of fumonisin level was observed in corn stored in closed conditions. The oils adversely affected kernel germination at 4.8 microL/g and therefore cannot be recommended for controlling F. verticillioides on stored corn used as seeds, when used at this concentration. The oil of neem seeds showed no inhibitory effect but rather accelerated the growth of F. verticillioides.

Life table study of Sitotroga cerealella (Lepidoptera: Gelichiidae), a strain from West Africa.

Life table studies for the Angoumois grain moth, Sitotroga cerealella (Olivier), a pest on stored maize, Zea mays L., in West Africa, were conducted as part of the expansion of a mathematical simulation model that has been developed for two pests of stored maize. The effects of four temperatures (20, 25, 30, and 35 degrees C) and two relative humidity levels (44 and 80%) on developmental time, age-specific survivorship and fecundity, sex ratio, and intrinsic rate of natural increase (r(m)) of S. cerealella were investigated. Sex ratio was close to 1:1 at all temperatures and humidity. Minimum development time occurred close to 32 degrees C and 80% RH for both males and females, and developmental time of females was significantly shorter than that of males. Immature survivorship was highest between 25 and 30 degrees C and 80% RH and lowest at 35 degrees C under both humidity conditions. A similar low level was found at 20 degrees C and 44% RH. The greatest fecundity (124 eggs per female) occurred at 20 degrees C, 80% RH. The maximum r(m) value was 0.086 d(-1) at 30 degrees C and 80% RH, but the growth rate declined dramatically at 35 degrees C. If compared with the few other life table studies conducted on this species on maize in India and North America, some variation among the strains becomes evident. A common conclusion for the current study and previous ones is that optimal population development for S. cerealella occurs at approximately 30 degrees C and at high humidity.

A comprehensive study to explore differences in mycotoxin patterns from agro-ecological regions through maize, peanut, and cassava products: a case study, Cameroon.

A total of 420 samples were collected from agrarian households. Whereas 51% (215/420) of the samples were contaminated with one or more toxins, the contamination rates for maize, peanut, and cassava products were 74, 62, and 24%, respectively. The fumonisins (20-5412 µg/kg), aflatoxin B1 (6-645 µg/kg), roquefortine C (1-181 µg/kg), and deoxynivalenol (27-3842 µg/kg) were the most prevalent contaminants in maize. For peanut samples, aflatoxin B1 (6-125 µg/kg) and ochratoxin A (0.3-12 µg/kg) were the main contaminants, whereas aflatoxin B1 (6-194 µg/kg) and penicillic acid (25-184 µg/kg) were detected in the cassava products. Exposures calculated through maize intake for fumonisin B1 and aflatoxin B1 were several-fold higher (2-5 for fumonisin B1 and 10(4)-10(5) for aflatoxin B1) than the health-based guidance values of 2 µg/kg bw/day and 0.15 ng/kg bw/day, respectively. The study design constitutes a good model that can be implemented in other sub-Saharan African countries.

Degradation of rotenone in yam bean seeds ( Pachyrhizus sp.) through food processing.

The purpose of this research is to screen different processes that could potentially decrease or even eliminate rotenone, a toxic isoflavonoid, from Pachyrhizus seeds. Yam bean seeds have very interesting nutritional characteristics, especially their high protein and lipid contents, and could potentially increase food security in under-nourished populations. However, they contain rotenone, a natural molecule previously used as an insecticide inhibiting the respiratory mitochondrial chain. It was also proven to be toxic to mammals as chronic exposure leads to the development of Parkinson-like symptoms in rats. As the thermosensitivity of rotenone had been reported, this study tested different processes (drying, roasting, boiling, frying, alcohol extraction), tegument removal, and traditional Beninese culinary recipes. Rotenone was then quantified in end-products by a validated method, associating microwave extraction, solid phase extraction (SPE), and HPLC-UV. With these processes a rotenone removal of up to 80% was obtained. The most effective methods were the drying and roasting of the seeds and the maceration of their flour in local alcohol. Rotenone degradation and elimination were confirmed by cytotoxic assays, effectively inducing a decrease in sample toxicity.