Multiple mycotoxins associated with maize (Zea mays L.) grains harvested from subsistence farmers' fields in southwestern Ethiopia.

Fifty-four maize grain samples freshly harvested from subsistence farmers' fields in southwestern Ethiopia were analyzed for multiple mycotoxins using liquid chromatography-tandem mass spectrometric (LC-MS/MS) method following extraction by acetonitrile/water/acetic acid on a rotary shaker. The grain samples were contaminated with a total of 164 metabolites, of which Fusarium and Penicillium metabolites were the most prevalent accounting for 27 and 30%, respectively. All the major mycotoxins and derivatives except one (citrinin) were of Fusarium origin. Zearalenone was the most frequent major mycotoxin occurring in 74% of the samples at concentrations of 0.32-1310 µg/kg. It was followed by nivalenol (63%), zearalenone-sulfate (44%), and fumonisin B1 (41%). Nivalenol, nivalenol glucoside, and fusarenon-X were detected at unusually high levels of 8-1700 µg/kg, 21-184 µg/kg, and 33-149 µg/kg, respectively. Deoxynivalenol and DON-3 glucoside contaminated 32% of the samples, each at levels of 15.9-5140 µg/kg and 10-583 µg/kg, respectively. Moniliformin and W493B occurred in 96 and 22% samples at levels of 3.27-4410 µg/kg and 3-652 µg/kg, respectively. Fumonisins were also detected in the samples at levels of 9-6770 µg/kg (B1), 16-1830 µg/kg (B2), 9.5-808 µg/kg (B3), and 1.3-128 µg/kg (A1). This study confirmed the presence of an array of mycotoxins contaminating maize grains right from the field. The effect of the co-occurring mycotoxins on consumers' health should be investigated along with that of the newly emerging ones. Results of the current study call for application of pre-harvest mycotoxin mitigation strategies to safeguard maize-based food and feed.

Cultural and morphological characteristics of Colletotrichum sublineolum isolates infecting sorghum in eastern Ethiopia.

Colletotrichum sublineolum is the most destructive pathogen causing sorghum anthracnose worldwide. The fungus is known to have highly variable pathotypes. A characteristic study of pathogen is important to document the change occurring in population as variability in morphology indicates the existence of different pathotypes. Controlled condition experiment was conducted to examine cultural and morphological characteristics of C. sublineolum isolates infecting sorghum in eastern Ethiopia. Sorghum leaves showing symptoms of anthracnose were collected from five districts through survey. To study the characteristics of C. sublineolum, single-spore isolates representing isolate collection districts were selected from the stock cultures and cultivated on potato dextrose agar. Culture growth, colony color, elevations, texture and margin, conidial diameter and shape were used to characterize isolates. The isolates were varied significantly in many aspects. Colony colors were differed from light-gray to gray, purple-gray to cottony-gray, white to salmon-whit, plum-pink to beige and rosy brown on upper side of the petri dishes. The mean culture growth of C. sublineolum isolates showed highly significant (P < 0.01) variations among each other and ranged from 15 to 44 mm eight-days after incubation. Most of the isolates were produced hyaline, smooth walled, falcate conidia but without septa. Conidial diameter of C. sublineolum isolates showed variations with width and length ranged from 2.97 to 6.01 µm and 10.01-27.75 µm, respectively. Most isolates had smooth colony margin and few had undulated margin. This finding revealed that substantial variations were observed among C. sublineolum isolates and the existence of variable characteristic showed the presence of several sub-species of the pathogen infecting sorghum in different agro-ecologies of eastern Ethiopia.

Key Global Actions for Mycotoxin Management in Wheat and Other Small Grains.

Mycotoxins in small grains are a significant and long-standing problem. These contaminants may be produced by members of several fungal genera, including Alternaria, Aspergillus, Fusarium, Claviceps, and Penicillium. Interventions that limit contamination can be made both pre-harvest and post-harvest. Many problems and strategies to control them and the toxins they produce are similar regardless of the location at which they are employed, while others are more common in some areas than in others. Increased knowledge of host-plant resistance, better agronomic methods, improved fungicide management, and better storage strategies all have application on a global basis. We summarize the major pre- and post-harvest control strategies currently in use. In the area of pre-harvest, these include resistant host lines, fungicides and their application guided by epidemiological models, and multiple cultural practices. In the area of post-harvest, drying, storage, cleaning and sorting, and some end-product processes were the most important at the global level. We also employed the Nominal Group discussion technique to identify and prioritize potential steps forward and to reduce problems associated with human and animal consumption of these grains. Identifying existing and potentially novel mechanisms to effectively manage mycotoxin problems in these grains is essential to ensure the safety of humans and domesticated animals that consume these grains.

Genetic fingerprinting and aflatoxin production of Aspergillus section Flavi associated with groundnut in eastern Ethiopia.

BACKGROUND: Aspergillus species cause aflatoxin contamination in groundnut kernels, being a health threat in agricultural products and leading to commodity rejection by domestic and international markets. Presence of Aspergillus flavus and A. parasiticus colonizing groundnut in eastern Ethiopia, as well as presence of aflatoxins have been reported, though in this region, no genetic studies have been done of these species in relation to their aflatoxin production. RESULTS: In this study, 145 Aspergillus isolates obtained from groundnut kernels in eastern Ethiopia were genetically fingerprinted using 23 Insertion/Deletion (InDel) markers within the aflatoxin-biosynthesis gene cluster (ABC), identifying 133 ABC genotypes. Eighty-four isolates were analyzed by Ultra-Performance Liquid Chromatography (UPLC) for in vitro aflatoxin production. Analysis of genetic distances based on the approximately 85 kb-ABC by Neighbor Joining (NJ), 3D-Principal Coordinate Analysis (3D-PCoA), and Structure software, clustered the isolates into three main groups as a gradient in their aflatoxin production. Group I, contained 98% A. flavus, including L- and non-producers of sclerotia (NPS), producers of B1 and B2 aflatoxins, and most of them collected from the lowland-dry Babile area. Group II was a genetic admixture population of A. flavus (NPS) and A. flavus S morphotype, both low producers of aflatoxins. Group III was primarily represented by A. parasiticus and A. flavus S morphotype isolates both producers of B1, B2 and G1, G2 aflatoxins, and originated from the regions of Darolabu and Gursum. The highest in vitro producer of aflatoxin B1 was A. flavus NPS N1436 (77.98 µg/mL), and the highest producer of aflatoxin G1 was A. parasiticus N1348 (50.33 µg/mL), these isolates were from Gursum and Darolabu, respectively. CONCLUSIONS: To the best of our knowledge, this is the first study that combined the use of InDel fingerprinting of the ABC and corresponding aflatoxin production capability to describe the genetic diversity of Aspergillus isolates from groundnut in eastern Ethiopia. Three InDel markers, AFLC04, AFLC08 and AFLC19, accounted for the main assignment of individuals to the three Groups; their loci corresponded to aflC (pksA), hypC, and aflW (moxY) genes, respectively. Despite InDels within the ABC being often associated to loss of aflatoxin production, the vast InDel polymorphism observed in the Aspergillus isolates did not completely impaired their aflatoxin production in vitro.

Multimycotoxin and fungal analysis of maize grains from south and southwestern Ethiopia.

The natural occurrence of fungi, mycotoxins and fungal metabolites was investigated in 100 samples of maize grains collected from south and southwestern Ethiopia in 2015. The maize samples were contaminated by Fusarium, Aspergillus and Penicillium species. Using liquid chromatography tandem mass spectrometry 127 secondary metabolites were analysed. Zearalenone was the most prevalent mycotoxin, occurring in about 96% of the samples. Zearalenone sulfate was the second most prevalent, present in 81% of the samples. Fumonisin B1 was detected in 70% of the samples with a mean level of 606 µg kg-1 in positive samples, while FB2, FB3 and FB4 were detected in 62%, 51% and 60% of the maize samples with mean levels of 202, 136 and 85 µg kg-1, respectively. Up to 8% of the samples were contaminated with aflatoxins, with a maximum level of aflatoxin B1 of 513 µg kg-1. Results were higher than earlier reports for maize from Ethiopia.

Morphological Variation and Inter-Relationships of Quantitative Traits in Enset (Ensete ventricosum (welw.) Cheesman) Germplasm from South and South-Western Ethiopia.

Enset (Ensete ventricosum (Welw.) Cheesman) is Ethiopia's most important root crop. A total of 387 accessions collected from nine different regions of Ethiopia were evaluated for 15 quantitative traits at Areka Agricultural Research Centre to determine the extent and pattern of distribution of morphological variation. The variations among the accessions and regions were significant (p ≤ 0.01) for all the 15 traits studied. Mean for plant height, central shoot weight before grating, and fermented squeezed kocho yield per hectare per year showed regional variation along an altitude gradient and across cultural differences related to the origin of the collection. Furthermore, there were significant correlations among most of the characters. This included the correlation among agronomic characteristics of primary interest in enset breeding such as plant height, pseudostem height, and fermented squeezed kocho yield per hectare per year. Altitude of the collection sites also significantly impacted the various characteristics studied. These results reveal the existence of significant phenotypic variations among the 387 accessions as a whole. Regional differentiations were also evident among the accessions. The implication of the current results for plant breeding, germplasm collection, and in situ and ex situ genetic resource conservation are discussed.

Aspergillus and aflatoxin in groundnut (Arachis hypogaea L.) and groundnut cake in Eastern Ethiopia.

This study was conducted to assess major Aspergillus species and aflatoxins associated with groundnut seeds and cake in Eastern Ethiopia and evaluate growers' management practices. A total of 160 groundnut seed samples from farmers' stores and 50 groundnut cake samples from cafe and restaurants were collected. Fungal isolation was done from groundnut seed samples. Aspergillus flavus was the dominant species followed by Aspergillus parasiticus. Aflatoxin analyses of groundnut seed samples were performed using ultra performance liquid chromatography; 22.5% and 41.3% of samples were positive, with total aflatoxin concentrations of 786 and 3135 ng g-1 from 2013/2014 and 2014/2015 samples, respectively. The level of specific aflatoxin concentration varied between 0.1 and 2526 ng g-1 for B2 and B1, respectively. Among contaminated samples of groundnut cake, 68% exhibited aflatoxin concentration below 20 ng g-1, while as high as 158 ng g-1 aflatoxin B1 was recorded. The study confirms high contamination of groundnut products in East Ethiopia.

Aflatoxin B1 and total fumonisin contamination and their producing fungi in fresh and stored sorghum grain in East Hararghe, Ethiopia.

Natural contamination of sorghum grains by aflatoxin B1 and total fumonisin and their producing toxigenic fungi has been studied. A total of 90 sorghum grain samples were collected from small-scale farmers' threshing floors and 5-6 months later from underground pits during 2013 harvest from three districts of East Hararghe, Ethiopia. Mycotoxin analysis was done using enzyme-linked immunosorbent assay (ELISA). The limits of detection were in the range 0.01-0.03 µg kg-1. The results revealed that all sorghum grain samples were contaminated with both Aspergillus and Fusarium species. Aflatoxin B1 was detected at levels ranging from