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  • About
  • The Global ETD Search service is a free service for researchers to find electronic theses and dissertations. This service is provided by the Networked Digital Library of Theses and Dissertations.
    Our metadata is collected from universities around the world. If you manage a university/consortium/country archive and want to be added, details can be found on the NDLTD website.
291

A study of the effects of storage methods on the quality of maize and household food security in Rungwe District, Tanzania.

Mboya, Rose. January 2011 (has links)
A sample of 260 farm households that were randomly selected in Katumba ward, Rungwe district, Tanzania were studied for the effects of storage methods on the quality of maize grain and household food security using qualitative and quantitative methods. Maize storage problems, amounts of maize that farm households harvested and amounts of maize that farm households lost to pests per year, food security status and farm households’ perceptions concerning their food security status were investigated using face - to - face semi - structured and structured interviews. Common storage methods that farm households used to store maize and the dietary importance of maize were investigated through interviews, seasonal calendars and the matrix for scoring and ranking. The quality of maize was investigated through conducting mycological analysis and through investigating levels of insect infestation using the incubation method on maize samples collected from a sub-sample of 130 farm households at harvest and after five months of storage period. It was found that farm households in Katumba ward preferred maize meal rather than other types of food that provide bulk such as rice and green bananas/plantains. Maize contributed 66.8 % - 69.5 % of the total energy and 83 - 90 % of the total protein required per day, and farm households stored maize using roof and sack storage methods. It was also found that 34.5 % of 2323 tonnes of maize that were harvested per annum in Katumba ward were lost to pests during storage. Fusarium, Diplodia, Aspergillus and Penicilliums species were identified as the main fungal pathogens that attacked stored maize. Sitophilus zeamais, Sitotroga cerealella and rodents were also identified as the main maize storage pests. About 25 % of the maize samples that were collected at harvest and 93 % of the maize samples that were collected from the same farm households after five months of storage were infested by either Sitophilus zeamais or Sitotroga cerealella or both. Maize samples from the two storage systems had an average number of 80 insect pests per 120 maize kernels (or 51 g of maize), amounting to 1569 insects per kg. The high levels of insect infestation reduced the amount of maize that could have been available to the farm households and subjected stored maize to fungal infections and subsequent contaminations, thus, rendering the farm households vulnerable to food insecurity. Furthermore, it was also found that most of the infestation of maize by insect pests and moulds in Katumba ward occurred during storage, and that farm households were not well informed concerning maize storage and the negative effects that fungal activities in maize can have on the health of the consumers. An average of 87717 μg/kg fumonisins, 596 μg/kg aflatoxins, 745 μg/kg ochratoxins and 1803 μg/kg T-2 toxins were detected in the maize samples. Currently, there are no set standards for T-2 toxins, whereas the internationally accepted standards for aflatoxins, fumonisins and ochratoxins in cereals are 20 μg/kg, 4 mg/kg and 50 μg/kg, respectively. It was concluded that the levels of mycotoxins detected in maize from Katumba ward were far above the internationally accepted standards and that the farm households were at risk of ill health through consuming maize meals made from contaminated maize grain. The presence of high concentrations of mycotoxins, together with the high levels of insect infestation in the maize led to the conclusion that reduction of the nutrient content of the maize grain in Katumba ward was inevitable. Thus, the pests that infested maize stored using the roof and sack storage methods in this ward compromised not only the availability of food, but also the utilization of the nutrients in the maize and its safety, leading to the farm households’ food insecurity. It was further concluded that the quality of maize stored using roof and sack storage methods in Katumba ward was low and that the roof and sack storage methods were inadequate for protecting stored maize from pests. It was recommended that an efficient method for rapid drying of maize prior to storage be found, that the roof and sack storage methods be improved so that they can effectively protect stored maize from moisture content problems. It was also recommended that the farm households’ awareness concerning maize storage and food security be raised, and that the extension staff in Katumba ward should urge the Tanzanian government to implement an agricultural policy which promotes efficient maize storage and maize quality in order to improve the current status quo. Above all, since maize is the predominant staple, it was recommended that the maize breeding program in Tanzania should emphasize development of maize varieties that are resistant to ear rots, storage insects and to contamination by mycotoxins as part of a larger program to improve food security in this part of the country. Breeding programs that aim at enhancing the nutritional value of maize were also recommended. / Thesis (Ph.D.)-University of KwaZulu-Natal, Pietermaritzburg, 2011.
292

Genetic improvement of Zambian maize (Zea mays L.) populations for resistance to ear rots and a survey of associated mycotoxins.

Mweshi, Mukanga. January 2009 (has links)
Maize ear rots are among the most important impediments to increased maize production in Africa. Besides yield loss, they produce mycotoxins in their host whose contamination has been linked to several human and animal mycoses. The main objectives of the studies reported on in this thesis were (i) to investigate farmer perceptions of maize ear rot disease and prospects for breeding for host plant resistance in Zambia; and (ii) to establish the levels of incidence and extent of maize ear rot infection as well as the level of mycotoxins in the maize crops of smallholder farms in central and southern Zambia; (iii) to appraise the field inoculation techniques and assess them for their suitability for the Zambian environmental conditions, (iv) to determine the combining ability of Zambian maize populations for resistance to ear rot and investigate the genetic basis of this resistance; and (v) to investigate both direct and indirect responses to full-sib selection for ear rot resistance in Zambian maize populations. A participatory rural appraisal (PRA) was conducted in four communities, involving a total of 90 farmers. Participatory methods were used, such as focused group discussions, group interviews, participant scoring and ranking. Farmers ranked and scored the various constraints affecting their maize production in general and the maize ear rots in particular. Ear rots were ranked as the third most important biotic stress and it was evident that although farmers were aware of the disease, they were not aware of mycotoxins. This was reflected in the way they disposed of rotten maize: either by feeding livestock or eating it in periods of hunger. The survey of ear rots and mycotoxins was carried out in the Southern and Central Provinces of Zambia. A total of 114 farms were covered in the survey: maize samples were collected and both ear rot fungi and mycotoxins were isolated. Fusarium and Stenocarpella were the most frequently isolated fungi from smallholder farms. The levels of fumonisins on these farms ranged from 0.05 to 192 ppm, while those of aflatoxins were between 1.5 and 10.6 ppb. In 50% of the farmsteads surveyed, the mycotoxins, i.e. fumonisins and aflatoxins, exceeded the recommended FAO/WHO 1limits of 2 ppm and 2 ppb, respectively. Five field inoculation techniques namely, colonised toothpick, leaf whorl placement, ear top placement, spore suspension spray, and silk channel injection, were evaluated over three seasons in a series of experiments. It was found that the leaf whorl placement of inoculums, followed by colonized toothpick method, gave a constant ranking of genotypes across locations and years compared to the other three methods. In addition, the use of a mixture of ear rots as inoculum was as effective as its principal single species constituents. In the population diallel analysis, five broad-based maize populations were crossed in a diallel and evaluated under artificial ear rot inoculation using an inoculum mixture of three ear rot fungi, Aspergillus flavus, Fusarium verticilloides and Stenocarpella maydis at four locations in Zambia. The purpose was to estimate general (GCA) and specific combining ability (SCA) and investigate genotype x environment interaction. GCA effects were found not to be significant for disease severity but were significant for grain yield across environments. Populations with a strong GCA effect for disease severity across sites included PRA783244c3, Pop25, MMV600, and ZUCASRc2. Across sites, the F1 combinations, MMV600 x Pop25, ZUCASRc2 X Pop25, and Pop25 x PRA783244c2 had strong SCA effects for root lodging, ear drooping, husk cover and ear insect damage. In a related diallel analysis of 10 full-sib families derived from these populations, it was observed that resistant x susceptible families and their reciprocal crosses performed better than their resistant parents, suggesting an over dominant expression of resistance. Both maternal and non maternal effects were observed to be influencing resistance to ear rots. There was a preponderance influence of non-additive gene action. A response to full-sib recurrent selection was conducted in four locations in Central Zambia. Out of the 343 families created in 2005/6 season, 10% were selected from each population and recombined to create five new populations. These, with the original populations, were evaluated in four sites during the 2007/8 season. There was a net reduction in ear rot incidence and rot severity in the new synthetic population. Pop10 had the largest reduction in disease severity. The predicted gain per cycle was -4.1% and realized gain was -2.5% for disease incidence, and 0.19% and 19.4% for grain yield. Genetic variability was maintained though with low heritability estimates. Negative but at times strong association between grain yield and ear rot disease severity was detected suggesting that in general selecting for ear rot resistance would enhance grain yield in the five populations. Overall the importance of the ear rots and mycotoxins in compromising yield and health of the communities in Zambia, respectively, were confirmed and support the call to improve maize varieties for resistance to ear rots. The results indicate that the five populations could be enhanced for ear rot resistance through population improvement procedures such reciprocal recurrent selection that exploit both additive and non-additive variation. Selection might be compromised by the large genotype x environment interaction effects, and large reciprocal effects and their interaction with the environments. To enhance repeatability genotypes should be artificially inoculated, by placing the inoculum in the leaf whorl followed by colonized toothpick inoculation, and screened in many environments to identify genotypes with stable resistance to ear rots. / Thesis (Ph.D) - University of KwaZulu-Natal, Pietermaritzburg, 2009.
293

Modulation of Aflatoxin B1 production by Aspergillus flavus / Modulation de la production d’Aflatoxine B1 chez Aspergillus flavus

Verheecke, Carol 25 November 2014 (has links)
Les mycotoxines sont des molécules toxiques produites par de nombreuses espèces fongiques. Les seules mycotoxines avérées aujourd’hui cancérigènes pour l’homme sont les aflatoxines. Elles sont produites par le genre Aspergillus principalement et sont retrouvées tout au long de la chaine alimentaire (champs, stockage, transformation, etc.). A cause du réchauffement climatique, la France devient de plus en plus exposée à la présence de ces mycotoxines. Afin de limiter l’exposition des consommateurs, de nombreuses stratégies de prévention ou de décontamination sont développées. Dans ce contexte, nous avons recherché à mettre au point un système de lutte biologique permettant de prévenir la production d’aflatoxines sur le maïs au champ. Pour cela, nous avons choisi des bactéries issues du sol et déjà connues pour être commercialisées pour la lutte biologique, les actinomycètes. Nous avons étudié l’interaction in vitro sur boites de Pétri entre Aspergillus flavus, principal producteur d’aflatoxines, et certains actinomycètes. Nous avons démontré que l’interaction peut réduire la concentration en aflatoxines mesurée par HPLC. De plus, certains isolats bactériens sont aussi capables de réduire, en culture pure, la concentration d’aflatoxine B1 dans le milieu. Des premiers tests d’adsorption ont été réalisés pour comprendre la nature de ce mécanisme. Par ailleurs, une étude approfondie via RT-qPCR sur 6 souches bactériennes du genre Streptomyces sp. A montré que celles-ci étaient capables d’impacter l’expression de différents gènes impliqués dans la voie de biosynthèse chez A. flavus et A. parasiticus. Enfin, nous avons complété les données déjà existantes sur l’impact de facteurs environnementaux (température, disponibilité en eau et du temps d’incubation) sur la production d’aflatoxines. / Mycotoxins are toxic contaminants of foodstuffs produced by a wide range of fungal species. Aflatoxins are the only mycotoxins carcinogenic for humans. They are mainly produced by the Aspergillus genus and can be found at each step of the agrofood chain (e.g. field, storage, process). Due to climate changes, France is starting to be exposed to aflatoxins. In order to limit the consumer exposure, many prevention or decontamination techniques have been developed. To this aim, we started the development of a biocontrol against aflatoxins accumulation for maize field application. Actinomycetes, are soil-borne bacteria that has already been commercialized as biocontrol. In Petri dishes, we studied the in vitro interaction between some actinomycetes and Aspergillus flavus, the main aflatoxins producer. We revealed that the interaction reduced the aflatoxins content (monitored by HPLC). Moreover, some bacterial isolates were able to reduce pure-aflatoxin B1 added in the medium. To understand this mechanism, adsorption tests has been conducted. Otherwise, RT-qPCR methodology was used to study the impact of Streptomyces-Aspergillus sp. on aflatoxin gene expression. Finally, the current knowledge of the impact of environmental factors (temperature, water activity and incubation time) on aflatoxins production was supplemented.
294

Base génétique et potentiel d’évolution de la pathogénicité de Fusarium graminearum, bio-agresseur fongique des céréales / Genetic basis and evolutionary potential of the pathogenicity of the fungus Fusarium graminearum

Laurent, Benoit 07 December 2016 (has links)
Le champignon Fusarium graminearum est l'un des principaux agents responsables de la fusariose des épis, une maladie nécrosante des céréales associée à une contamination des grains et des aliments par des mycotoxines. De récentes observations suggèrent une évolution de l’agressivité des populations de ce pathogène, questionnant l’efficacité et la durabilité des moyens de luttes actuels. Mieux anticiper cette évolution nécessite une meilleure caractérisation de la diversité phénotypique et génotypique existante entre souches. Six nouveaux génomes de F. graminearum ont été séquencés et ont permis l’identification et la caractérisation de 243 000 variations génétiques. La majorité de ces variants (77%) est concentrée dans des îlots de polymorphisme, représentant 32% du génome et enrichis en probables effecteurs liés à la pathogénicité de F. graminearum. La construction d’une population recombinante, et son génotypage avec 1 300 marqueurs moléculaires, ont permis le développement de la première carte génétique à haute-densité de l’espèce. La corrélation entre le taux de recombinaison et le polymorphisme a mis en évidence une organisation « à deux-vitesses » du génome de cette espèce. Finalement, l’intégration de ces données dans une approche de génétique quantitative a permis l’identification d’un locus polymorphe, affectant le gène FgVeA, et responsable de 90% de la variation d’agressivité et de la production de mycotoxine observée. Les différents résultats obtenus durant ces travaux font l’objet d’une discussion générale sur le potentiel adaptatif et d’évolution de ce pathogène. / F. graminearum is one of the main causal agents of the fusarium head-blight (FHB), a cereal disease leading to head necrosis, in addition to grain and food/feed contamination by stable and toxic metabolites. Recent observations refer to an increase of pathogenicity, questioning efficiency and durability of current management practices. In order to anticipate this evolution, we must bring a deeper characterization of the currently existing diversity. Six new genomes of F. graminearum were sequenced, and 243,000 genetic variations have been identified and characterized. Seventy seven percent of the total number of the variants was located within 32% of the genome, delineating highly polymorphic islands. These islands are enriched with probable effectors linked to Fusarium’s pathogenicity. The construction and the genotyping on 1,300 molecular markers of a recombinant population have enabled the development of the first high-density genetic map of the species. The remarkable correlation between polymorphism and recombination rate highlighted the 'two-speed' genome organization of this pathogen. Finally, the integration of these data through a quantitative genetic approach allowed the discovery of one quantitative trait locus, likely to affect the gene FgVeA, and responsible for 90% of the observed variation of aggressiveness and mycotoxin production. These results are discussed in the light of F. graminearum’s adaptive potential and evolution.
295

Impact des mycotoxines sur le microbiote intestinal humain, cas particulier du déoxynivalénol / Impact of mycotoxins on the human gut microbiota, particular case of deoxynivalenol

Saint-Cyr, Manuel 18 December 2013 (has links)
Le déoxynivalénol (DON) est une mycotoxine qui contamine la plupart des cultures de céréales dans toutes les régions du monde. Capable de résister aux procédés de transformation subies par les céréales, le DON peut se retrouver alors, à l'état de contaminants dans les matières premières (céréales) ainsi que dans les denrées alimentaires transformées destinées à l'Homme (pâtes, pain, bières) et à l'animal (granulés) à des concentrations supérieures aux limites règlementaires. Malgré les efforts de recherche pour caractériser les multiples aspects de l’impact d’une contamination par le DON, les effets bactériologiques de cette mycotoxine n’étaient pas encore documentés chez l’Homme. L'Agence Nationale de Sécurité Sanitaire (Anses), dans le cadre de sa mission de protection du consommateur, a donc souhaité évaluer l'impact d'une contamination au DON sur le microbiote intestinal humain (MIH). Dans cette étude, nous avons d’abord évalué la cinétique du DON chez le porc et chez le rat, puis nous avons utilisé un modèle de rats à flore humanisée pour évaluer l'impact d'une exposition sub-chronique de la mycotoxine sur la composition du MIH. Le DON est un contaminant rapidement distribué et éliminé. Au sein du tractus digestif, il entraine des changements bactériologiques significatifs chez certains principaux groupes bactériens composant le MIH. Cette étude apporte des données complémentaires à l’analyse du risque lié à l’exposition du DON chez l’Homme et montre l’intérêt des modèles animaux étudiés dans des scénarii particuliers d’exposition au DON. / Deoxynivalenol (DON) is one of the most prevalent mycotoxins present in cereal crops worldwide. Able to withstand the transformation process undergone by grains, DON can be found as contaminant in raw materials (cereals) and in processed food for humans (pasta, bread, beer) and animals (grains) at concentrations upper the control limits. Despite research efforts to characterize various aspects of the impact of DON contamination, the microbiological effects of DON were not documented in humans. The French agency for food, environmental and occupational health and safety (Anses), as part of its mission to protect the consumer, wanted to assess the impact of DON contamination on the Human Gut Microbiota (HGM). In this study, we first evaluated the kinetics of DON in pigs and rats, and then we assessed the impact of an oral subchronic exposure of deoxynivalenol on the composition of HGM in a human microbiota-associated rats model. DON is a contaminant rapidly distributed and eliminated. In gut, DON leads to significant changes in some of the main bacterial groups of the HGM. This study provides additional data to analyze the risk exposure of DON in humans and shows the interest of these animal models in studies dealing with particular scenarios of DON exposure.
296

HUMAN AND ANIMAL HEALTH RISK ASSESSMENT OF MYCOTOXIN MIXTURES IN MAIZE: FROM FUNGAL PRODUCTION AND OCCURRENCE TO HARMONISED RISK CHARACTERISATION

PALUMBO, ROBERTA 03 April 2020 (has links)
Maize is the principal staple food/feed crop exposed to mycotoxins, and the co-occurrence of multiple mycotoxins and their metabolites has been well documented. Dietary (co)-exposure to mycotoxins is associated with human and animal health concerns as well as economic losses. The present thesis aims to apply a holistic approach for the risk assessment of mycotoxin mixtures in food and feed, i.e. from fungal production and occurrence to harmonised risk characterisation. This was done in three folds. Firstly, available environmental, ecological, and agronomic factors that may affect the relative abundance of co-occurring mycotoxins in the contaminated crops were collected from peer-reviewed literature, with focus on maize (Chapter I). Secondly, (co-)occurrence data on mycotoxins in core cereals was extracted from available articles in the scientific literature and analysed to estimate potential pattern of co-exposure in humans and animals (Chapter II). Finally, Chapter III investigates the applicability of the EFSA guidance to multiple mycotoxins through a scenario of possible co-exposure in humans and animals, using maize as a case study. In particular, a human and animal risk assessment to mycotoxin mixture in maize was conducted using a modelled component-based approach for selected mixture of mycotoxins, that, according to our data, co-occur in maize based feed and food products. / Maize is the principal staple food/feed crop exposed to mycotoxins, and the co-occurrence of multiple mycotoxins and their metabolites has been well documented. Dietary (co)-exposure to mycotoxins is associated with human and animal health concerns as well as economic losses. The present thesis aims to apply a holistic approach for the risk assessment of mycotoxin mixtures in food and feed, i.e. from fungal production and occurrence to harmonised risk characterisation. This was done in three folds. Firstly, available environmental, ecological, and agronomic factors that may affect the relative abundance of co-occurring mycotoxins in the contaminated crops were collected from peer-reviewed literature, with focus on maize (Chapter I). Secondly, (co-)occurrence data on mycotoxins in core cereals was extracted from available articles in the scientific literature and analysed to estimate potential pattern of co-exposure in humans and animals (Chapter II). Finally, Chapter III investigates the applicability of the EFSA guidance to multiple mycotoxins through a scenario of possible co-exposure in humans and animals, using maize as a case study. In particular, a human and animal risk assessment to mycotoxin mixture in maize was conducted using a modelled component-based approach for selected mixture of mycotoxins, that, according to our data, co-occur in maize based feed and food products.
297

Využití hmotnostní spektrometrie ke stanovení markerů oxidativního stresu a mykotoxinů / Application of Mass Spectrometry for the Determination of Oxidative Stress Markers and Mycotoxins

Čumová, Martina January 2015 (has links)
The first topic presented in the dissertation thesis is determination of isoprostanes as markers of oxidative stress and other compounds affected by presence of oxidative stress. Isoprostanes iPF2-III, iPF2-VI, iPF2-VI, astaxanthin and polyunsaturated fatty acids (PUFA), especially arachidonic acid (AA) were monitored in Atlantic salmon eggs (Salmo salar). Methods for the determination of these compounds have been developed and optimized using chromatographic separation coupled to conventional or mass spectrometric detection. Freshly laid eggs, eyed embryos and non-viable eggs were used to test a general hypothesis that egg viability can be affected by susceptibility to oxidative stress, either through the specific fatty acid concentration and/or the antioxidant capacity of the eggs. Levels of isoprostanes and arachidonic acid (AA) were significantly higher in non-viable eggs than in control (eyed embryos) as well as relative abundance of PUFA. While no difference of isoprostanes was found between freshly laid and control those from the Atlantic stock except iPF2-VI which was observed under the LOQ in the control. Higher levels of PUFA and AA in comparison with the control were observed in the freshly laid eggs. However, the only statistically significant difference was observed in the amount of astaxanthin. Different levels of PUFA and astaxanthin may be related to their biochemical consumption during the development of eggs. This work evaluated potential effect on the viability of eggs Salmo salar due to the presence of oxidative stress. The monitoring of mycotoxins in food and feed was the subject of the second topic. Mycotoxins are secondary metabolites produced by fungi. They are ubiquitous undesirable natural contaminants that are toxic for humans and animals. Today are known more than 500 mycotoxins. However, only few of them are regulated by the European Union. The European Food Safety Authority (EFSA) was asked by the European Commission to provide a scientific opinion on other mycotoxins for which statutory limits could be developed. In this study is proposed simultaneous screening allowing fast, reliable and sensitive approach, identification and quantification of 17 mycotoxins in food and feed sample. The method includes both mycotoxins regulated by the EU and selected mycotoxins required by the EFSA (aflatoxins, deoxynivalenol, nivalenol, zearalenone, fumonisin, ochratoxin A, T-2 toxin, HT-2 toxin, enniatins and beauvericin). Analytes are isolated by the modified QuEChERS method. For separation and target mycotoxins detection, ultra-performance liquid chromatography coupled to tandem mass spectrometry (UPLC –MS/MS) was employed. The method also allows determination of ergot alkaloids (ergocornine, ergosine, ergocryptine, ergocristine and their respective epimers). The developed method was used either for monitoring mycotoxins and ergot alkaloids in feed and raw materials and barley and malt prepared from it.
298

Estimating mycotoxin exposure and increasing food security in Guatemala

Garsow, Ariel V. January 2022 (has links)
No description available.
299

Microbiota and mycotoxins in traditional beer of the greater Kimberley area and associated brewing and consumption practices

Ikalafeng, Bridget Keromamang January 2008 (has links)
Thesis (D. Tech.) -- Central University of Technology, Free State, 2008 / The purpose of this study was to evaluate brewing and consumption practices and to screen for micro-organisms and mycotoxins associated with traditional beer produced and consumed in the marginal urban settlements of the city of Kimberley in the Northern Cape Province of South Africa. The survey study revealed that traditional beer is no longer being brewed for traditional purposes only, as was the case in the past, but rather for commercial gain. Both brewers and consumers, however, appeared to be largely unaware of disease-causing micro-organisms present on the hands or bodies of handlers that can be transferred to the beverage during the handling process, and were seemingly not conversant with regard to the effects of hazardous ingredients sometimes incorporated during the brewing process. Unemployment and a lack of education emerged as pivotal factors related to the production of traditional beer and the ignorance of the associated safety thereof. The survey further indicated that although facilities such as the availability of potable water (taps in yards) and flushing toilets were sometimes in place, other facilities such as basins with hot running water were often not available. In commercially produced and homebrewed traditional beer the mean counts for total coliforms and Staphylococcus spp. were circa 105 cfu.ml-1 whereas the TVC (Total Viable Counts) and total fungi counts were 106 and 107 cfu.ml-1 respectively. The total coliforms and Staphylococcus spp. counts for homebrewed traditional beer were approximately one log-phase higher than the commercial version. The counts in the homebrewed beer probably originated from contamination during handling, while in the commercial product contamination originated either in the raw ingredients or during postprocessing and consumption. Apart from staphylococci, considerable numbers of total coliforms indicating faecal contamination were noted. A rapid, easy, reliable and accurate technique that could be used to quantify the level of mycotoxins (deoxynivalenol and citrinin) in the beer was developed through validation of the ELISA Ridascreen methodology. Using this method, the deoxynivalenol (DON) level in the beer samples was found to exceed the recommended levels suggested by the European Union, while citrinin levels in the samples varied between 35.6 ppb and 942.2 ppb. In the case of citrinin there were statistically significant differences between spring, summer and winter samples, confirming the seasonal impact on fungal growth and consequent mycotoxin production. An R2-value of 0.409 was noted between DON and citrinin, indicating a weak positive association. Finally, an awareness programme in the format of a poster with accompanying subscripts was developed to address issues of safety and hygiene of traditional beer in the study area. The poster utilises animatedstyle colour images of selected practices that need to be addressed, accompanied by slogans summarising the particular image in English, Afrikaans and Setswana. It is envisaged that, as part of a comprehensive awareness programme, the poster will contribute greatly to the quality, safety and promotion of traditional beer in the area.
300

Molecular interactions of arbuscular mycorrhizal fungi with mycotoxin-producing fungi and their role in plant defense responses

Ismail, Youssef 11 1900 (has links)
Les trichothécènes de Fusarium appartiennent au groupe des sesquiterpènes qui sont des inhibiteurs la synthèse des protéines des eucaryotes. Les trichothécènes causent d’une part de sérieux problèmes de santé aux humains et aux animaux qui ont consommé des aliments infectés par le champignon et de l’autre part, elles sont des facteurs importants de la virulence chez plantes. Dans cette étude, nous avons isolé et caractérisé seize isolats de Fusarium de la pomme de terre infectée naturellement dans un champs. Les tests de pathogénicité ont été réalisés pour évaluer la virulence des isolats sur la pomme de terre ainsi que leur capacité à produire des trichothécènes. Nous avons choisi F. sambucinum souche T5 comme un modèle pour cette étude parce qu’il était le plus agressif sur la pomme de terre en serre en induisant un flétrissement rapide, un jaunissement suivi de la mort des plantes. Cette souche produit le 4,15-diacétoxyscirpénol (4,15-DAS) lorsqu’elle est cultivée en milieu liquide. Nous avons amplifié et caractérisé cinq gènes de biosynthèse trichothécènes (TRI5, TRI4, TRI3, TRI11, et TRI101) impliqués dans la production du 4,15-DAS. La comparaison des séquences avec les bases de données a montré 98% et 97% d'identité de séquence avec les gènes de la biosynthèse des trichothécènes chez F. sporotrichioides et Gibberella zeae, respectivement. Nous avons confrenté F. sambucinum avec le champignon mycorhizien à arbuscule Glomus irregulare en culture in vitro. Les racines de carotte et F. sambucinum seul, ont été utilisés comme témoins. Nous avons observé que la croissance de F. sambucinum a été significativement réduite avec la présence de G. irregulare par rapport aux témoins. Nous avons remarqué que l'inhibition de la croissance F. sambucinum a été associée avec des changements morphologiques, qui ont été observés lorsque les hyphes de G. irregulare ont atteint le mycélium de F. sambucinum. Ceci suggère que G. irregulare pourrait produire des composés qui inhibent la croissance de F. sambucinum. Nous avons étudié les patrons d’expression des gènes de biosynthèse de trichothécènes de F. sambucinum en présence ou non de G. irregulare, en utilisant le PCR en temps-réel. Nous avons observé que TRI5 et TRI6 étaient sur-exprimés, tandis que TRI4, TRI13 et TRI101 étaient en sous-exprimés en présence de G. irregulare. Des analyses par chromatographie en phase-gazeuse (GC-MS) montrent clairement que la présence de G. irregulare réduit significativement la production des trichothécènes par F. sambucinum. Le dosage du 4,15-DAS a été réduit à 39 μg/ml milieu GYEP par G. irregulare, comparativement à 144 μg/ml milieu GYEP quand F. sambucinum est cultivé sans G. irregulare. Nous avons testé la capacité de G. irregulare à induire la défense des plants de pomme de terre contre l'infection de F. sambucinum. Des essais en chambre de croissance montrent que G. irregulare réduit significativement l’incidence de la maladie causée par F. sambucinum. Nous avons aussi observé que G. irregulare augmente la biomasse des racines, des feuilles et des tubercules. En utilisant le PCR en temps-réel, nous avons étudié les niveaux d’expression des gènes impliqué dans la défense des plants de pommes de terre tels que : chitinase class II (ChtA3), 1,3-β-glucanase (Glub), peroxidase (CEVI16), osmotin-like protéin (OSM-8e) et pathogenèses-related protein (PR-1). Nous avons observé que G. irregulare a induit une sur-expression de tous ces gènes dans les racines après 72 heures de l'infection avec F. sambucinum. Nous avons également trové que la baisse provoquée par F. sambucinum des gènes Glub et CEVI16 dans les feuilles pourrait etre bloquée par le traitement AMF. Ceci montre que l’inoculation avec G. irregulare constitut un bio-inducteur systémique même dans les parties non infectées par F. sambucinum. En conclusion, cette étude apporte de nouvelles connaissances importantes sur les interactions entre les plants et les microbes, d’une part sur les effets directs des champignons mycorhiziens sur l’inhibition de la croissance et la diminution de la production des mycotoxines chez Fusarium et d’autre part, l’atténuation de la sévérité de la maladie dans des plantes par stimulation leur défense. Les données présentées ouvrent de nouvelles perspectives de bio-contrôle contre les pathogènes mycotoxinogènes des plantes. / Fusarium trichothecenes are a large group of sesquiterpenes that are inhibitors of eukaryotic protein synthesis. They cause health problems for humans and animals that consume fungus-infected agricultural products. In addition some of Fusarium trichothecenes are virulence factors of plant pathogenesis. In this study, sixteen Fusarium strains were isolated and characterized from naturally infected potato plants. Pathogenicity tests were carried out to evaluate the virulence of these isolates on potato plants and their trichothecene production capacity. We chose F. sambucinum strain T5 as a model for this study because it was the most aggressive strain when tested on potato plants. It induces a rapid wilting and yellowing resulting in plant death. This strain produced 4,15-diacetoxyscirpenol (4,15-DAS) when grown in liquid culture. We amplified and characterized five trichothecene genes (TRI5, TRI4, TRI3, TRI11, and TRI101) involved in the production of 4,15-DAS. Nucleotide BLAST search showed 98% and 97% sequence identity with trichothecene biosynthetic genes of F. sporotrichioides and Gibberella zeae, respectively. We used F. sambucinum to determine if trichothecene gene expression was affected by the symbiotic arbuscular mycorrhizal fungus (AMF) Glomus irregulare. We found that the growth of F. sambucinum was significantly reduced in the presence of G. irregulare isolate DAOM-197198 compared with controls that consisted of carrot roots without G. irregulare or F. sambucinum alone. Furthermore, inhibition of the growth F. sambucinum was associated with morphological changes, which were observed when G. irregulare hyphae reached F. sambucinum mycelium, suggesting that G. irregulare may produce compounds that interfere with the growth of F. sambucinum. Using real-time qRT-PCR assays, we assessed the relative expression of trichothecene genes of F. sambucinum confronted or not with G. irregulare. When G. irregulare was confronted with F. sambucinum, TRI5 and TRI6 genes were up-regulated, while TRI4, TRI13 and TRI101 were down-regulated. We therefore used GC-MS analysis to determine whether G. irregulare affects trichothecene production by F. sambucinum. We found that the production of 4,15-DAS trichothecene was significantly reduced in the presence of G. irregulare compared with controls that consisted of carrot roots without G. irregulare or F. sambucinum alone. Interestingly, 4,15-DAS pattern was reduced to 39 μg/ml GYEP medium by G. irregulare compared to 144 μg/ml GYEP with F. sambucinum grown with carrot roots or F. sambucinum alone respectively. We tested the AMF capacity to induce defense responses of potato plants following infection with F. sambucinum. The response of AMF-colonized potatoes to F. sambucinum was investigated by tracking the expression of genes homologous with pathogenesis-related proteins chitinase class II (ChtA3), 1,3-β-glucanase (gluB), peroxidase (CEVI16), osmotin-like protein (OSM-8e) and pathogenesis-related protein (PR-1). We found that the AMF treatment up-regulated the expression of all defense genes in roots at 72 hours post-infection (hpi) with F. sambucinum. We also found that a decrease provoked by F. sambucinum in gluB and CEVI16 expression in shoots could be blocked by AMF treatment. Overall, a differential regulation of PR homologues genes in shoots indicates that AMF are a systemic bio-inducer and their effects could extend into non-infected parts. In conclusion, this study provides new insight into on the interactions between plants and microbes, in particular the effects of AMF on the growth and the reduction of mycotoxins in Fusarium. It also shows that AMF are able to reduce the disease severity in plants by stimulating their defense. The data presented provide new opportunities for bio-control against mycotoxin-producing pathogens in plants.

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