Mycotoxins are toxic secondary metabolites produced by filamentous fungi. Aflatoxins and fumonisins are among the most toxic mycotoxins. They are a significant risk factor for a cocktail of chronic health conditions including cancer of the liver, oesophagus and kidney, teratogenicity, neural tube defects, interference with lipid metabolism, a weakened immune system and a negative impact on micronutrient absorption in both man and animals. This study compared natural contamination of peanuts, peanut butter and sorghum from Gaborone, Botswana and Bulawayo, Zimbabwe with aflatoxins and fumonisins. In total 34 peanut samples, 34 sorghum samples and 11 peanut butter samples were collected randomly from retail shops and informal markets in the two cities. Fungal contamination was determined using standard mycology methods. Aflatoxin and fumonisin contamination was determined using HPLC-FLD. A. flavus/parasiticus species were detected in 66% and 100% of randomly analysed peanut samples from Bulawayo and Gaborone respectively and 27% (3/11) of peanut butter samples from Bulawayo. 67% of randomly analysed sorghum samples from Bulawayo showed A. flavus/parasiticus and Fusarium species contamination while none of the randomly analysed sorghum samples from Gaborone showed any fungal contamination. Furthermore aflatoxins were not detected in any of the sorghum samples; however 61% (11/18) of the Bulawayo sorghum samples showed fumonisin contamination (Range: 8 – 187 ng/g). Three of the peanut samples from Bulawayo were contaminated with aflatoxins (range: 6.6 – 622 ng/g) and no aflatoxins were detected in Gaborone peanuts. All 11 peanut butter samples from Bulawayo were contaminated with aflatoxins (Mean: 73.5 ng/g, Range: 6.8-250 ng/g) and AFB1 was the most prevalent. These preliminary results indicate that peanut butter and peanuts from Bulawayo are contaminated with high levels of aflatoxins. Stricter policing of regulations should be implemented to ensure compliance by manufacturers and public health interventions implemented in vulnerable communities. / Life & Consumer Sciences / M. Sc. (Life Sciences)

Mycotoxins are toxic secondary metabolites produced by filamentous fungi. Aflatoxins and fumonisins are among the most toxic mycotoxins. They are a significant risk factor for a cocktail of chronic health conditions including cancer of the liver, oesophagus and kidney, teratogenicity, neural tube defects, interference with lipid metabolism, a weakened immune system and a negative impact on micronutrient absorption in both man and animals. This study compared natural contamination of peanuts, peanut butter and sorghum from Gaborone, Botswana and Bulawayo, Zimbabwe with aflatoxins and fumonisins. In total 34 peanut samples, 34 sorghum samples and 11 peanut butter samples were collected randomly from retail shops and informal markets in the two cities. Fungal contamination was determined using standard mycology methods. Aflatoxin and fumonisin contamination was determined using HPLC-FLD. A. flavus/parasiticus species were detected in 66% and 100% of randomly analysed peanut samples from Bulawayo and Gaborone respectively and 27% (3/11) of peanut butter samples from Bulawayo. 67% of randomly analysed sorghum samples from Bulawayo showed A. flavus/parasiticus and Fusarium species contamination while none of the randomly analysed sorghum samples from Gaborone showed any fungal contamination. Furthermore aflatoxins were not detected in any of the sorghum samples; however 61% (11/18) of the Bulawayo sorghum samples showed fumonisin contamination (Range: 8 – 187 ng/g). Three of the peanut samples from Bulawayo were contaminated with aflatoxins (range: 6.6 – 622 ng/g) and no aflatoxins were detected in Gaborone peanuts. All 11 peanut butter samples from Bulawayo were contaminated with aflatoxins (Mean: 73.5 ng/g, Range: 6.8-250 ng/g) and AFB1 was the most prevalent. These preliminary results indicate that peanut butter and peanuts from Bulawayo are contaminated with high levels of aflatoxins. Stricter policing of regulations should be implemented to ensure compliance by manufacturers and public health interventions implemented in vulnerable communities. / Life and Consumer Sciences / M. Sc. (Life Sciences)

Philosophiae Doctor - PhD / Immunosensors are affinity ligand-based biosensor solid-state devices in which the immunochemical reaction is coupled to a transducer. The specificity of the molecular recognition of antigens by antibodies to form a stable complex is the basis of the immunosensor on the electrode. The development of such a sensor requires a better design and preparation of an optimum interface between the biomolecules and the detector material. The immunosensors were developed based on Polyaniline derivative composite. Novel water soluble PdTe quantum dots (QD) was synthesized and characterized by different physical techniques such as UV-Visible (UV-VIS), Fluorescence Spectroscopy (PL), Scanning Electron Microscopy (SEM), Transmission Electron Microscopy (TEM) and X-Ray Diffraction (XRD). The electroactivity of such synthesized quantum dots was studied by cyclic voltammetry in aqueous media. The synthesis of poly(2,5- dimethoxyaniline)-multi wall 'carbon nanotubes nanocomposite was carried out by electropolymerization in situ of 2,5-dimethoxyaniline - multi wall carbon nanotubes (PDMA-MWCNT) from aqueous dispersion containing acid-treated multi wall carbon nanotubes (MWCNT) and 2,5-dimethoxyaniline subsequently modifying a glassy carbon electrode in acid media. An undoped PDMA was also prepared for control. The composite for this work, consists of layer-by-layer method to form a multilayer film of QDs and PDMA-MWCNT. The method used was as follows; the drop coating of quantum dots followed by electrodeposition of poly(2,5- dimethoxyaniline )-carbon nanotubes onto surface of glassy carbon. The PDMA-CNT was characterized by UV-Visible (UV-Vis), Fourier Transform Infrared Spectroscopy (FTIR) and Scanning Electron Microscopy (SEM). The electrochemical characterisation of PDMA-CNT was carried out using cyclic voltammetry (CV) and Electrochemical Impedance Spectroscopy (EIS). The composite (QDs-PDMA-MWCNT) was also characterized using above mentioned techniques. The electrochemical immunosensor for fumonisin a mycotoxin was prepared by dropcoating of mycotoxins antibody onto the composite modified glassy carbon electrode. The response profiles of fumonisins sensors system were obtained from electrochemical impedance spectroscopy (EIS) measurements. The fumonisin immunosensor was used for the detection of fumonisins in certified com reference materials. For comparison reasons, analysis of such mycotoxins was carried out by using conventional analytical method enzyme-linked immunosorbent assay (ELISA). The EIS response of FBI immunosensor (GCEIPT-PDMA-MWCNT/anti-Fms-BSA) gave a linear range of 7 to 49 ng L-I and the corresponding sensitivity and detection limits were 0.0162 ka L ng-I and 0.46 pg L-I, respectively. Hence the limit of detection of GCEIPT-PDMA-MWCNT immunosensor for fumonisins in com certified material was calculated to 0.014 and 0.011 ppm for FBI, and FB2 and FB3, respectively. These results are lower than those obtained by ELISA, a provisional maximum tolerable daily intake (PMTDI) for fumonisins (the sum of FBI, FB2, and FB3) established by the Joint FAO / WHO expert committee on food additives and contaminants of 2 ug kg" and the maximum level recommended by the U.S. Food and Drug Administration (FDA) for protection of human consumption (2-4 mg L-I).

Thesis (MSc)--University of Stellenbosch, 2000. / ENGLISH ABSTRACT: Please see fulltext for abstract / AFRIKAANSE OPSOMMING: Sien asb volteks vir opsomming

Thesis (MScAgric (Plant Pathology)--Stellenbosch University, 2008. Includes bibliography.summary,list of figures and appendices. / ENGLISH ABSTRACT: Fusarium spp. and Aspergillus spp. are toxin-producing fungi associated with maize and groundnut. Fusarium verticillioides produces fumonisins in maize, and Aspergillus flavus produces aflatoxins in maize and groundnut kernels. Both toxins are responsible for carcinogenesis in humans and animals. Contamination of maize and groundnut with mycotoxins is often most severe in rural areas where subsistence farmers are unaware of their existence and follow agricultural practices that might contribute to their production. A questionnaire was, therefore, compiled to investigate agricultural decisions in rural areas that may influence mycotoxin contamination of crops. During 2006 and 2007, maize and groundnut samples were collected in the Eastern Cape, KwaZulu-Natal (KZN), Limpopo, and Mpumalanga provinces. Mycotoxin levels were quantified using the ELISA technique, and the incidence of Fusarium spp. in maize grain was determined by plating maize kernels out on Fusarium selective medium. Fumonisin-producing Fusarium spp. were also quantified using real-time PCR (TaqMan). The incidence of A. flavus and A. parasiticus in groundnut was determined by plating out kernels on potato dextrose agar. Fumonisin contamination levels in maize samples ranged from 0-21.8 parts per million (ppm) and aflatoxin levels ranged from 0- 49 parts per billion (ppb), depending on the region where samples were collected. Aflatoxin levels in groundnut ranged from 0-160.1 ppb. Fusarium verticillioides was the most common Fusarium sp. in maize followed by F. subglutinans and F. proliferatum, respectively. Regression analyses showed a positive correlation between fumonisin-producing Fusarium species when determined by real-time PCR and fumonisin concentration (r2=0.866). Regression analyses further showed a highly significant positive correlation between A. flavus and aflatoxin contamination (r2=0.10235). Samples from northern KZN contained levels of mycotoxins that were far in excess of the maximum levels set by the Food and Drug Administration in the USA. In South Africa there are currently no regulations with regard to the maximum allowable levels of fumonisin in human food. The high incidence of mycotoxin contamination of human food in subsistence farming systems indicates the need for awareness programmes and further research. / AFRIKAANSE OPSOMMING: Fusarium spp. en Aspergillus spp. is toksien-produserende fungi wat met mielies en grondbone geassosieer word. Fusarium verticillioides produseer fumonisiene in mielies, terwyl, A. flavus aflatoksiene in mielies en grondbone produseer. Beide toksiene is karsinogenies vir mens en dier. Die vlakke van toksien-kontaminasie is meestal die ergste in landelike gebiede waar bestaansboere onbewus is daarvan. Landboupraktyke wat deur die boere toegepas word vererger dikwels die probleem. ‘n Vraelys is saamgestel om vas te stel watter landboupraktyke in landelike gebiede toegepas word, en hoe dit toksien-kontaminasie in mielies en grondbone beinvloed. In die 2006 en 2007 seisoene is mielie- en grondboonmonsters in Kwa-Zulu-Natal (KZN), die Oos Kaap, Limpopo en die Mpumalanga provinsie versamel. Toksien-vlakke is gekwantifiseer deur gebruik te maak van die ELISA tegniek. Die insidensie van Fusarium spp. in mielies was bepaal deur pitte op Fusariumselektiewe agar uit te plaat. Fumonisien-produserende Fusarium spp. was ook gekwantifiseer deur van kwantitatiewe PCR (TaqMan) gebruik te maak. Die voorkoms van A. flavus en A. parasiticus is bepaal deurdat mielie- en grondboonpitte op aartappel dekstrose agar uit te plaat. Fumonisien-vlakke in die mieliemonsters het gewissel van 0-21.8 dele per miljoen (dpm), terwyl aflatoksienvlakke gewissel het van 0-49 dele per biljoen (dpb), afhangende van die omgewing waar monsters versamel is. Aflatoksien vlakke in die grondboonmonsters het gewissel van 0-160.1 dpb. Fusarium verticillioides is die meeste vanuit mielies geisoleer, gevolg deur F. subglutinans en F. proliferatum. Regressie analises het ‘n positiewe korrelasie tussen fumonisien konsentrasie en fumonisien-produserende spp. aangedui waar daar gebruik gemaak is van die kwantitatiewe PCR (r2= 0.866). Regressie analises het ‘n hoogs betekenisvolle positiewe korrelasie getoon tussen A. flavus en aflatoksien kontaminasie (r2=0.10). Monsters van noordelike KZN het toksienvlakke bevat ver bokant die maksimum toelaatbare vlakke is soos bepaal deur die Food en Drug Administrasie in die Verenigde State van Amerika. Daar is tans geen regulasies in Suid Afrika wat die maksimum toelaatbare vlakke van fumonisiene in voedsel vir menslike gebruik bepaal nie. Die hoë voorkoms van mikotoksien-kontaminasie in bestaansboer-sisteme, dui die belangrikheid van verdere navorsing en bewusmakings-progamme aan.

Thesis (DSc (Biochemistry))--Stellenbosch University, 2009. / The fumonisin mycotoxins are known to be the causative principle for several animal diseases and are associated with the development of liver and oesophagus cancer and neural tube defects in humans. The thesis focuses mainly on the characterisation of the compounds from maize cultures of the fungus Fusarium verticillioides, isolated from maize, the toxicological effects in animals, mechanism involved in hepato- and nephrocarcinogenicity and discussing the major differences and contradictions in the literature together with their impact on setting relevant risk assessment parameters to safeguard human health. Controversies include the importance of non-genotoxicity vs genotoxicity in the development of cancer, the role of threshold effects in carcinogenesis and the establishment of realistic risk assessment parameters that will also be applicable in developing countries. Recent approaches suggest that thresholds should also apply for genotoxic carcinogens as interaction with the DNA is only one event in the multi-step process of cancer development and therefore could not be taken as the basis for applying a no-effect threshold for genotoxins. It would appear that a carcinogen such as fumonisin, whether it is labeled genotoxic or non-genotoxic per se, exhibits some degree of risk at any level due to additive or synergistic interactions with other xenobiotics and/or dietary constituents. The underlying mechanisms of fumonisin-induced carcinogenicity includes the disruption of sphingolipid, phospholipids and fatty acid metabolism, which plays a major role in the modulation of apoptotic and cell proliferative pathways related to cancer development. Interactive responses between arachidonic acid and ceramide affect downstream cell signal transduction pathways and depending on the cell type the disruption of these pathways could either stimulate or inhibit cell proliferation which eventually will determine the induction of apoptosis and hence affect cell survival. The modulating roles of dietary constituents such as vitamins, protein and the South African herbal teas are also highlighted as they affected the outcome of toxicological assays, thus determining thresholds of the adverse effects in specific target organs that will impact risk assessment parameters. Regulation of the fumonisins in food and the associated risk are debated from many perspectives. In developing countries there is a lack of quality control implying that maize highly contaminated with mycotoxins may directly enter the food chain of adults and children as control of mycotoxins is difficult or in some cases totally absent. The interaction of politics, economy and technology will eventually determine the impact on health as the regulation of fumonisin in food differs between countries. Knowledge about the biological effects of the fumonisins is currently playing an important role in the development of simple and inexpensive methods to reduce the levels of the fumonisin in maize by targeting specific populations at risk.

Made available in DSpace on 2016-04-28T20:17:26Z (GMT). No. of bitstreams: 1 2006 - Rodrigo Alcantara de Carvalho.pdf: 803403 bytes, checksum: 7a35f34784a3f9cb58b545f52959a6a0 (MD5) Previous issue date: 2006-09-22 / The world wide contamination of foods and byproducts with mycotoxins is a problem of extreme importance. Mycotoxins are secondary metabolites produced by fungi that have adverse effects to human and animal health. The fumonisins and aflatoxins are the mycotoxins of greatest agro-economic importance due to the damage brought to animal production. Important steps in the synthesis of these mycotoxins are described, as such as their metabolism and adverse effects. This work evaluates the determination of the biomarkers sphinganin (Sa) and sphingosin (So) in poultry serum as a diagnose method for fumonisin exposition, and compares the obtained data with ration analysis. Of the evaluated animals only 49% exhibited 2 sphingolipids (Sa and So), 53,04% exhibited only So and 2,04% exhibited only Sa. It was detected at ration the presence of fumonisin B1 at concentration of 374,92μg.kg-1 and aflatoxins B1, B2, and G1 at concentrations of 440,99μg.kg-1, 169,92μg.kg-1 and 494,58μg.kg-1, respectively. It was possible to partially establish the grade of intoxication of the poultry and to verify that the sensibility of the biomarker can be affected by the presence of fumonisin such as aflatoxins. / A contamina??o mundial de alimentos e subprodutos com micotoxinas ? um problema de extrema import?ncia. As micotoxinas s?o metab?litos secund?rios produzidos por fungos, que possuem efeitos adversos ? sa?de humana e animal. As fumonisinas e aflatoxinas s?o as micotoxinas de maior import?ncia agro-econ?mica pelos preju?zos acarretados ? produ??o animal. Passos importantes na s?ntese destas micotoxinas s?o descritos, assim como seu metabolismo e efeitos adversos. Este trabalho avalia a determina??o dos biomarcadores esfinganina (Sa) e esfingosina (So) no soro de frangos de corte como m?todo de diagn?stico para exposi??o ?s fumonisinas e compara os dados obtidos com a an?lise da ra??o. Dos animais avaliados apenas 49% exibiram quantidades detect?veis de esfingolip?deos no soro, dentre estes, apenas 44,9% exibiram os 2 esfingolip?deos (Sa e So), 53,06% exibiram somente So e 2,04% exibiram somente Sa. Foi detectada na ra??o a presen?a de fumonisina B1 na concentra??o de 374,92μg.kg-1 e de aflatoxinas B1, B2 e G1 nas concentra??es de 440,99μg.kg-1, 169,92μg.kg-1 e 494,58μg.kg-1, respectivamente. Foi poss?vel estabelecer parcialmente o grau de intoxica??o dos frangos e verificar que a sensibilidade do biomarcador pode ser afetada tanto pela presen?a de fumonisina quanto de aflatoxina.

In questa tesi di dottorato sono stati indagati i punti critici del ciclo vitale del fungo Fusarium verticillioides, produttore di fumonisine in mais. Le fumonisine sono metaboliti secondari prodotte da funghi appartenenti tutti al genere fusarium e sono state classificate come sostanze cancerogene per l'uomo e per gli animali. Gli argomenti trattati nei vari capitoli sono stati: contaminazione da fumonisine nel mais coltivato nel nord Italia ed effetto delle diverse tecniche colturali, produzione di spore, dispersione di spore, infezione, parametri ecologici che condizionano la crescita e l'accumulo di fumonisina ed il controllo con mezzi chimici in campo su F. verticillioides e Ostrinia nubilalis, un lepidottero che aumenta la contaminazione. / Aim of this work was to collect missing information about critical point of the disease life cycle, to develop a conceptual model for the dynamic simulation of the life cycle of Fusarium verticillioides in maize and the consequent production of fumonisin B1 in kernels. Fumonisin is a secondary metabolite reported as toxigenic in humans and animals. Issues treated are: Fumonisin contamination in maize in Italy and effect of field managements, spore production, spore dispersal, variables influencing growth and toxin accumulation and chemical control of the disease in field.

Aflatoxin B1 (AFB1) and Fumonisin B1 (FB1), potentially cytotoxic and carcinogenic mycotoxins are common contaminants of agricultural commodities in South Africa and thus could be detrimental to the human immune system. Many of the cytotoxic effects of AFB1 require its bioactivation to an epoxide, which will bind covalently to macromolecules to form protein and DNA adducts. Fumonisin B1 is a competitive inhibitor of sphingosine and sphinganine N aceyltransferase, which are key components in the pathways for sphingolipid biosynthesis. Accumulation of free sphingoid bases, which are both cytotoxic and mitogenic, could provide a plausible explanation for the toxicity and carcinogenicity of FB1. The cytotoxic effects of AFB1 and FB1 on normal human lymphocytes, individually and in combination were assessed using the methylthiazol tetrazolium (MTT) bioassay. Two different methods of treatment were used, the treatment of isolated normal human lymphocytes for 12, 24, 48, 72 and 96 hours and whole blood treated for 12 hours. Flow cytometry and fluorescent microscopy were used to determine whether AFB1 and FB1 (5uM and 50uM), individually or in combination, were capable of inducing apoptosis, necrosis or nuclear fragmentation in isolated lymphocytes and whole blood treated for 12 hours. DNA damage was evaluated using the comet assay. The results showed that AFB1routinely induced higher levels of cytotoxicity in isolated lymphocytes than FB1. In the combination treatment, the mitogenic properties of FB1 appeared to partially counteract the cytotoxic effect exerted by AFB1. When whole blood was treated with the same concentration and ratio of toxin, FB1 was shown to be more cytotoxic than AFB1. The combination treatment of whole blood was shown to be cytotoxic in a dose dependent manner. The toxins appeared to exert a greater cytotoxic effect, when treated in combination than individually at higher concentrations. Aflatoxin B1 induced increased levels of apoptosis and necrosis in isolated lymphocytes while treatment with the FB1 resulted in increased levels of apoptosis at both concentrations. Treatment with the combination also resulted in increased levels of apoptosis. The levels of apoptosis were reduced in whole blood lymphocytes when compared to isolated lymphocytes. However, treatment with AFB1 and FB1 resulted in increased levels of apoptosis. Both AFB1 and FB1 are capable of inducing nuclear fragmentation. Treatment with FB1 (5uM and 50uM) resulted in greater degree of fragmentation than AFB1. The most nuclear fragmentation was induced by the 5uM combination treatment. The 50uM combination treatment of isolated lymphocytes induced the most DNA damage. As both toxins are common contaminants and have been known to coexist, this could be a potential area of concern for public health. / Thesis (M.Med.)-University of KwaZulu-Natal, 2004.

The use of the multi-mycotoxin screen based on dialysis to analyze foods and feeds for mycotoxins, is well documented. This study investigated the possibility of incorporating FB I into the screen. Maize meal (25g) was spiked with AFB I , CPA, FB1, ST and ZEA and extraction was done using acetonitrile/4% potassium chloride (90:10 v/v). The recoveriesof the mycotoxins were 77.4, 61.5, 97.4, 79.8 and 98% respectively on analysis by HPLC. Fumonisin B1 could not be completely incorporated into the screen due to its reaction with sodium hydrogen carbonate, which is a component in the method. Thus, FB I was determined in a separate portion of the extract. The high cost of FBI standards which are often of inferior purity necessitated that FB I standards be locally produced in the laboratory using Fusarium moniliforme MRC 826, a good producer of FB 1 . In this study, production of FB I was carried out using a stirred jar fermenter and patty cultures. The yields were 160mg/1 and 6mg/g of FB I for the two methods respectively. Methyl esterification of tricaballylic acid moieties of FB I was done for effective clean-up. This was achieved by derivatizing FBI, with diazomethane. It was found that other functional groups besides the tricaballylic acid moieties of FB I were undesirably methylated as well, which made cleanup by this method difficult as shown by electrospray mass spectrometric analysis. Attempts to de-methylate FBI methyl esters with esterase was not successful. Analysis of human faecal samples was carried out with the view of developing a short term marker for assessing human exposure to FB I . Faeces from rural (20) and urban (23) volunteers were analyzed by high performance liquid chromatography. The results showed that 35% of the rural samples and 9% of the urban volunteers had detectable amounts of FB I ranging from 0.600 to 19.56 mg/kg. There was a significant difference (p = 0.04)between the two population groups. A study was carried out to assess the occurrence of FBI in a rural area of Tugela valley in Kranskop magisterial district of KwaZulu Natal. A questionnaire was administered to gather information on the family health and nutrition. Raw (stored) and processed foods and faeces, were collected for analysis of FB1. A similar control study was carried out in the urban area of Durban Metro. Homes were mapped out using the GIS for easy follow up. Oesphageal cancer (OC) incidence from the local hospital and weather data for the study area were collected from South African Weather Bureau, Johannesburg. The questionnaire results showed that the common diseases were mainly of respiratory origin (24% and 26%) from both rural and urban groups respectively. Food analysis (by HPLC) showed that the number of maize samples with FB I were higher in the rural area (31.9%) in comparison to the urban samples (6.1%). The level ranged from 0.092-22.225 mg/kg in food and 0.513-39 mg/kg in faeces. The mean concentration of FB i in the faeces and maize samples showed a similar significant difference of 0.014 between the two groups. However, these concentrations were much lower than those of high OC area in Transkei (117 mg/kg). There was no detection of FBI in fermented food products. / Thesis (M.Med.Sci.)-University of Natal, Durban, 1998.