Samuel M. Imathiu

Molecular studies to identify the Fusarium species responsible for HT-2 and T-2 mycotoxins in UK oats

High levels of Fusarium mycotoxins HT-2 and T-2 have been detected in UK oats since surveys started in 2002. Fusarium langsethiae and the closely related species F. sporotrichioides have previously been associated with the contamination of cereals with type A trichothecenes HT-2 and T-2 in Nordic countries. Preliminary microbiological analysis of UK oat samples with high concentrations of HT-2 and T-2 detected and isolated F. langsethiae and F. poae but not the other type A trichothecene producing species F. sporotrichioides, F. sibiricum and F. armeniacum. Two hundred and forty oat flour samples with a known mycotoxin profile were selected from a previous four year study (2002-2005) to cover the full concentration range from below the limit of quantification (<20 μg/kg) to 9,990 μg/kg HT-2+T-2 combined. All samples were analysed for the DNA of F. langsethiae, F. poae and F. sporotrichioides based on previously published PCR assays. F. langsethiae was detectable in nearly all samples; F. poae was detected in 90% of samples whereas F. sporotrichioides was not detected in any sample. A real-time PCR assay was developed to quantify F. langsethiae DNA in plant material. The assay could quantify as low as 10(-4)ngF. langsethiae DNA/μl. Based on this assay and a previously published assay for F. poae, both species were quantified in the oat flour samples with known HT-2+T-2 content. Results showed a good regression (P<0.001, r(2)=0.60) between F. langsethiae DNA and HT-2+T-2 concentration. F. poae DNA concentration was not correlated to HT2+T2 concentration (P=0.448) but was weakly correlated to nivalenol concentration (P<0.001, r(2)=0.09). Multiple regression with F. langsethiae and F. poae DNA as explanatory variates identified that both F. langsethiae and F. poae DNA were highly significant (P<0.001) but F. poae DNA only accounted for an additional 4% of the variance and the estimate was negative, indicating that higher concentrations of F. poae DNA were correlated with slightly lower concentrations of HT2+T2 detected. A stronger regression (P<0.001, r(2)=0.77) between F. langsethiae DNA and HT-2+T-2 was obtained after extraction and quantification of DNA and mycotoxins from individual oat grains. The results from this study provide strong evidence that F. langsethiae is the primary, if not sole, fungus responsible for high HT-2 and T-2 in UK oats.

Fusarium langsethiae pathogenicity and aggressiveness towards oats and wheat in wounded and unwounded in vitro detached leaf assays

In vitro detached leaf assays involving artificial inoculation of wounded and unwounded oat and wheat leaves were used to investigate the potential pathogenicity and aggressiveness of F. langsethiae, which was linked recently to the production of type A trichothecenes, HT-2 and T-2 in cereals in Europe. In the first two experiments, two assays compared disease development by F. langsethiae with known fusarium head blight pathogen species each used as a composited inoculum (mixture of isolates) at 10°C and 20°C and found all fungal species to be pathogenic to oat and wheat leaves in the wounded leaf assay. In the unwounded leaf assay, F. langsethiae was not pathogenic to wheat leaves. Furthermore, there were highly significant differences in the aggressiveness of pathogens as measured by lesion length (Pu2009<u20090.001). In the second two experiments, pathogenicity of individual F. langsethiae isolates previously used in the composite inoculum was investigated on three oat and three wheat varieties. The wounded leaf assay showed that all isolates were pathogenic to all oat and wheat varieties but only pathogenic towards oat varieties in the unwounded assay. Highly significant differences (Pu2009<u20090.001) in lesion length were found between cereal varieties as well as between isolates in the wounded assay. Significant differences in lesion lengths (Pu2009=u20090.014) were also observed between isolates in the unwounded assay. Results from the detached leaf assays suggest that F. langsethiae is a pathogen of wheat and oats and may have developed some host preference towards oats.

Evaluation of pathogenicity and aggressiveness of F. langsethiae on oat and wheat seedlings relative to known seedling blight pathogens

Fusarium and Microdochium species are causal agents of seedling blight of small-grain cereal crops where they may contribute to a significant reduction in crop establishment and final yield. Two experiments were carried out to investigate the potential pathogenicity and aggressiveness of F. langsethiae, a recently identified fungus linked with the contamination of cereals with high levels of the trichothecene mycotoxins, HT-2 and T-2. An artificial seed inoculation method involving conidial suspensions was used and the experiments conducted in a growth cabinet set at either 5 or 15°C with a 12xa0h photoperiod. Known seedling blight pathogens of the genus Fusarium and Microdochium were used for comparison. At 15ºC, F. culmorum, M. nivale and M. majus caused seedling blight of oats and wheat with F. culmorum, on average being the most aggressive than the latter two. At 5ºC, only F. culmorum and M. nivale caused seedling blight of oats and wheat. Under the experimental conditions employed, F. langsethiae and F. poae failed to produce seedling blight disease indicating that these two species are not pathogenic to oat and wheat cultivars, Gerald and Claire respectively, at the seedling stage of development. They are therefore unlikely to affect crop establishment and other yield components such as tiller number, grain yield per head as well as grain weight if there is no subsequent foot-rot and/or head blight where infected seeds are sown.

Low permeability triple-layer plastic bags prevent losses of maize caused by insects in rural on-farm stores

Participatory on-farm trials were conducted to assess effectiveness of Purdue Improved Crop Storage (PICS™) bags for storage of maize in small-scale farmers’ stores in rural villages in eastern Kenya. A PICS bag is a three-layered hermetic bag-system that forms a barrier against the influx of oxygen and the escape of carbon dioxide. Jute, woven polypropylene or PICS bags were filled with shelled maize grain, purchased from the participating farmers, and the three sets of bags kept in the farmers’ own stores for 35xa0weeks. Oxygen and carbon dioxide levels in the PICS bags were monitored, as well as the temperature and relative humidity in all the bags. Grain moisture, live insect population, grain damage and weight loss were examined at intervals of seven weeks. Oxygen and carbon dioxide composition demonstrated that PICS bags are capable of sustaining good air-barrier properties under farmer storage conditions. Moreover, moisture content of maize stored in PICS bags did not change throughout the storage period whereas the moisture content of maize stored in polypropylene and jute bags decreased significantly in the final 14xa0weeks. Maize stored in PICS bags remained free from insect infestation and the weight loss due to insect damage was below 1xa0%. On the contrary, polypropylene and jute bags permitted profuse build-up of insect populations. At 35xa0weeks, grain damage reached 77.6xa0% and 82.3xa0% corresponding to 41.2xa0% and 48.5xa0% weight loss in the polypropylene and jute bags respectively. These findings demonstrate that PICS bags are effective in controlling losses caused by storage pests under farmer storage conditions.

In vitro growth characteristics of Fusarium langsethiae isolates recovered from oats and wheat grain in the UK

Fusarium langsethiae is a fungus that has recently been implicated in the contamination of small-grain cereal crops such as oats, wheat and barley with high levels of HT-2 and T-2 toxins in many European countries. The epidemiology of this fungus is not well known and may therefore be a bigger problem than currently thought to be. A study was carried out investigating the in vitro growth characteristics of F. langsethiae isolates from contaminated oats and wheat at various temperatures; 15, 20, 25 and 30 °C. Results indicated similar growth trends of oats and wheat isolates of F. langsethiae. Wheat isolates grew significantly (p<0.001) faster than oat isolates although this difference may have been confounded by the age of cultures, with oat isolates collected one year earlier. The estimated optimum growth temperature for all isolates was 24 °C. Isolates were macro-morphologically categorized as having lobed or entire colony margins, and either possessing one of the following colony colours: white, orange or purple. Since the estimated optimum growth temperature of F. langsethiae is typical in temperate summers when small-grain cereals are flowering, it is possible that this species can infect, colonise and possibly contaminate the developing grains with HT-2 and T-2 toxins which are of food safety concern.

Evaluation of the proximate composition, antioxidant potential, and antimicrobial activity of mango seed kernel extracts

Abstract After pulp extraction in fruit processing industry, a significant quantity of mango seed kernels are discarded as solid wastes. These seed kernels can be ideal raw materials for obtaining extracts rich in bioactive compounds with good antioxidant properties. The conversion of these wastes into utilizable food ingredients would help in reducing environmental problems associated with processing waste disposal. In order to determine their potential use, this study evaluated some of the biochemical characteristics and antimicrobial potential of mango seed kernel extracts on medically important human bacterial and fungal pathogens. Four mango varieties (Apple, Ngowe, Kent and Sabine) from Makueni and Embu counties in Kenya were used for this study. The analyzed mango seed kernel powders were found to contain on average, 6.74–9.20% protein content. Apple and Ngowe mango seed kernels had significantly higher fat content of 13.04 and 13.08, respectively, while Sabine from Makueni had the least fat content of 9.84%. The ash, fiber, and carbohydrate contents ranged from 1.78 to 2.87%, 2.64 to 3.71% and 72.86 to 75.92%, respectively. The mean percentage scavenging ability of mango kernel extracts at the concentration of 20 mg/mL was 92.22%. Apple and Sabine mango kernel extracts had significantly high inhibition zones of 1.93 and 1.73 compared to Kent and Ngowe with 1.13 and 1.10, respectively, against E. coli. For C. albicans, the inhibition of Kent mango kernel extract, 1.63, was significantly lower than that of Ngowe, Apple, and Sabine with 2.23, 2.13, and 1.83, respectively. This study demonstrates that mango seed powder is an abundant and cost‐effective potential natural antibiotic and antifungal that can be utilized in addressing the challenge of food poisoning and infections caused by pathogenic microorganisms in the food industry.

Agronomic practices influence the infection of an oats cultivar with Fusarium langsethiae

Fusarium langsethiae, a toxigenic fungus known to contaminate small-grain cereals with type A trichothecene mycotoxins, HT-2 and T-2 was described as a new species in 2004. HT-2 and T-2 are some of the most potent Fusarium toxins in eukaryotes, capable of inhibiting protein synthesis. The epidemiology of F. langsethiae is not well understood and with the intent of the European Commission to set maximum levels of contamination of cereals with these toxins, importance is currently placed in trying to understand the fungal infection process and its favorable growth conditions. A field study was carried out to investigate the effect of artificially inoculated oats straw, ploughing and minimum tillage with and without incorporated crop debris (straw) on infection and mycotoxin production by F. langsethiae on oats cultivar Gerald. The results indicated that cultural field practices had effects on the infection of oats by F. langsethiae. Fusarium langsethiae DNA was quantified in significantly larger amounts (p 0.05) on oat infection by F. langsethiae as quantified by DNA concentration. HT-2+T-2 quantification and analysis, gave no good evidence that either inoculation or cultural practice had any significant influence on the concentration of mycotoxins in the samples (p>0.05), but samples from minimum tillage with incorporated straw plots resulted in 2.5 times more HT-2+T-2 toxins than samples from ploughed with removed straw. These findings indicate the importance of tillage and crop debris management in the mitigation in an effort to prevent F. langsethiae infection, colonization and possible contamination of oats with HT-2 and T-2 toxins.

Effects of Varying Grilled Sorghum Content on the Quality Parameters of Urwagwa, a Traditional Rwandese Banana-based Alcoholic Beverage

An alcoholic beverage called Urwagwa is a common and popular traditional Rwandese drink that is produced from banana juice supplemented with coarsely ground grilled sorghum. Due to limited information on how different sorghum concentration affected the physico-chemical characteristics of this traditional beverage, a study was carried out to investigate how varying this ingredient influenced the quality of the final product. Different levels of grilled coarsely ground sorghum (0%, 2%, 5%, 7%, 10% and 15%) were mixed with banana juice and natural fermentation allowed to take place in six separate fermenters for six days at room temperature (25°C). Four parameters; ethanol content, total soluble solids, pH and titratable acidity were monitored after every 24 hours for 144 hours. Results obtained at the end of fermentation indicated that the ethanol content in all fermenters ranged from 10-12% v/v, total soluble solids 8.1-9.7°Brix, pH 4.0-4.7 and titratable acidity 0.6-0.9%. The highest fermentation rate was observed in fermenters containing ground grilled sorghum compared to the control (0% sorghum added).The highest total soluble solid at the completion of fermentation was found in banana beverage with zero percent of sorghum while the highest pH was generally found in fermenters with sorghum combinations. High titratable acidity was observed in fermenter with zero percent of sorghum. The concentration of 5% of ground grilled sorghum was found to result to the most desirable alcoholic banana beverage as it gave the highest yield of ethanol content and other biochemical parameters, as well as a more desirable flavor. Based on these findings, it could be recommended that the traditional Urwagwa brewers adopt the 5% of sorghum content in the production process in order to obtain a better quality and more acceptable alcoholic beverage.

The effect of different processing methods on nutrient and isoflavone content of soymilk obtained from six varieties of soybean grown in Rwanda

Abstract Soymilk is rich in nutrients and isoflavones, and could greatly promote nutrition and health. However, this product is not widely accepted due to an objectionable beany flavor. Several methods involving heat treatment and soaking in basic solutions prior to soymilk extraction have been reported to reduce the objectionable flavor. However, the effects of such treatments on the nutritional value and isoflavone content of soymilk, and the responses of different soybean varieties to nutrient extraction by these methods is not well studied. The aim of this study was to determine the effect of three processing methods on protein, fat, minerals, and isoflavone content in soymilk from six soybean varieties grown in Rwanda (Peka‐6, SB 24, Sc. Sequel, Sc, Squire, and a local variety) to find the best variety and processing method. The first method (M1) involved soaking soybeans in water for 12 hr prior to milk extraction, M2 involved blanching in NaHCO 3 prior to extraction and M3 involved soaking in NaHCO 3 solution for 16 hr and subsequent cooking prior to extraction. M1 resulted in significantly higher nutrient and isoflavone extraction than M2 and M3. Thus, M1 extracted more nutrients and can be recommended for soymilk production. However, where consumers prefer soymilk obtained by M2 or M3, Sc Squire and the local variety may be recommended. Sc. Squire has another advantage of higher isoflavone content than the other varieties. Further comprehensive studies on the sensory acceptability of products made from different varieties by different methods among different consumer categories will be necessary.

Effects of pretreatment during drying on the antioxidant properties and color of selected tomato varieties

Abstract Drying is essential in lowering the water activity and increasing the shelf stability of perishables. Thus, this study investigated the effect of pretreatment on the retention of the antioxidant properties and color of four tomato varieties; that is, Anna F1, Kilele, Prostar F1, and Riogrande during drying. Prepared quarters were treated by spraying with 0.5% sodium metabisulfate, 0.5% calcium chloride, and distilled water. The quarters were oven dried at 50°C, 60°C, and 70°C to 13% moisture content. Lycopene, β carotene, total phenolics, color, and moisture content were determined in both the fresh and dried samples. Initial moisture content among the four varieties did not differ significantly and ranged between 94.2 and 94.6%. Results showed that the main effects were significant (p < .05) on all measurable variables. Significantly (p < .05) higher retention levels in lycopene, β carotene, total phenolics, and lightness was observed in chemically pretreated samples compared to the control during drying.