Theses and dissertations (Applied Sciences)
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Item Incidence and characterization of Fusarium species in crown rot of bananas(2002) Ramsunder, Kumindra Devrajh; Odhav, Bharti; Okole, Blessed N.Fusarium species produce toxic mycotoxins that are known to exert adverse health effects in humans and animals. No attempts have been made to establish mycotoxin-producing capabilities of isolates of Fusarium species from bananas exhibiting symptoms of crown rot. Crown rot is one of the most serious post harvest problems in banana and the disease is caused by different fungal species, principally Fusarium species. Banana, which is of great economic significance in growing countries (i.e. Costa Rica, Cameroon, Ecuador) is seriously affected by crown rot and is a major cause of fruit lossItem The isolation and characterization of phytoalexin and constitutive agents from plants for mycotoxin control(2000) Mohanlall, Viresh; Odhav, BhartiPlant medicine is an important area of commercial activity in South Africa. This is a rapidly expanding market, thus we are evaluating natural and stressinduced compounds (phytoalexins) from plants as agents that may be able to control mycotoxins. Natural compounds from Bridelia micrantha, Warburgia salutaris, Lippia javanica and Scenecio serratuloides and stress-induced compounds (phytoalexins) from Citrus sinensis cv Valencia were screened for antitunqal and antimycotoxic activity by bioautography against a test organism (Cladosporium cladosporoides) and mycotoxin producing fungi (Fusarium moniliforme and Aspergillus flavus).Item The potential of spice oils in the control of mycotoxin producing fungi(2000) Juglal, Sarla; Odhav, BhartiSpice oils are known to exhibit antifungal activity and therefore have the potential to control mycotoxin production. There is a need in the food industry to find measures to control mycotoxins that are frequently associated with grains that form the staple diet of the majority of the population in South Africa. Clove, cinnamon, oregano, tumeric, eucalyptus, neem, aniseed, mace and nutmeg oils were tested to determine their inhibitory potential against growth of Aspergillus parasiticus and Fusarium moniliforme using the agar overlay technique. Varying concentrations of the spice oils, ranging from 0.1 ppm to 2.0 ppm, were incorporated into broth cultures of A. parasiticus and maize patty cultures ofF. moniliforme. Levels of production of aflatoxins and fumonisin were determined using standard thin layer chromatography and highpressure liquid chromatography methods. In addition, the active component of the spice oils were isolated, characterised and tested. The inhibitory potential of these compounds for field use was tested by incorporating clove oil, whole cloves and ground cloves in sampItem Partial characterization of toxigenic Fusarium(2004) Govender, Leroosha; Govinden, R.Various methods have been developed for the analysis of Fusarium and its toxins. Advances in molecular biology can lead to efficient characterization of this group of fungi. This study was undertaken to examine random amplified polymorphic DNA, volatile compound production and hydrolytic enzyme production by 19 Fusarial isolates. These techniques were employed to assess their abilities in differentiating Fusarium species and F. verticillioides strains and extending the analysis to discriminate toxin producing capabilities amongst these fungiItem Concurrent analysis of the mycotoxins, cyclopiazonic acid, moniliformin and ochratoxin A using capillary zone electrophoresis(2000) Govender, Urishani; Odhav, BhartiMycotoxins are a group of natural poisons produced by certain strains of fungal species when they grow under favourable conditions on a wide variety of different substrates. These toxins have been implicated in a wide range of acute diseases in man and animals. Their toxic effects include oesophageal cancer and liver diseases in humans, and carcinogenic effects in experimental rats and poultry. Hence, there is a need to monitor toxin levels in food commodities.Item Modulating effects of Fumonisin B1 and Ochratoxin A on immune cells in human carcinoma(2005) Adam, Jamila Khatoon; Odhav, BhartiFumonisin B1 (FB1) and ochratoxin A (OTA) represent examples of mycotoxins of greatest public health and agro-economic significance. They exert adverse effects on humans, animals and crops that result in illnesses and economic losses. Fumonisin B1 are cancerpromoting metabolites of Fusarium proliferatum and F verticillioides, (formerly moniliforme), and are implicated in oesophageal cancer. Ochratoxins are metabolites of both Aspergillus and Penicillium species. These compounds are known for their nephrotoxic effects in all animal species and may promote tumours in humans. In man OTA exhibits unusual toxicokinetics, with a half-life in blood of 840 h (35 days) after oral ingestion. Although much is known regarding the toxicology of these toxins, little is known of the effects of these toxins on the immune system. The aim of this study was to determine and compare the immunornodulating effects of FB1 and OTA in human carcinoma. Initial experiments involved isolating lymphocytes and neutrophils from healthy volunteers. The isolated cells were exposed to either FB1 or OTA on a dose and time dependent level and LD50 of the toxins was determined. Thereafter, challenge tests were performed, whereby lymphocytes and neutrophils isolated from volunteers, oesophageal cancer patients and breast cancer patients were exposed to the LD50 dose of either FB1 or OTA for the appropriate time. The effect of the toxins was demonstrated by viability studies, light microscopy and electron microscopy. Cytokine receptors (CK, TNF and CSF) were evaluated by immuno-cytochemical methods and the levels of circulating cytokines (IL -1, IL-6, IL-8, IL-10 and TNF-a) were determined using ELISA kits.Item Molecular characterization of aflatoxigenic and non-aflatoxigenic aspergillus isolates(2007) Mngadi, Phakamile TruthFor decades the genus Aspergillus (of fungi) has been classified based on morphological and growth criteria. Members of the Aspergillus section Flavi are economically valuable and methods of differentiating them are thus very important. Several molecular methods have been developed to distinguish these strains. Also, a number of biochemical and genetic studies have been used in order to provide a better means of classification (Lee et al., 2004). Aflatoxins, the most frequently studied mycotoxins, are produced by certain Aspergillus species/strains/isolates of fungi. The aflatoxin biosynthetic pathway studies have led to a number of discoveries. Several structural and regulatory genes (and their enzymes) involved in the biosynthesis of aflatoxins have been discovered and purified (Trail et al., 1995). Aflatoxin production and contamination of agricultural crops are major causes of economic losses in agriculture. Thus, better methods of characterization/differentiation are required for both aflatoxigenic and non-aflatoxigenic isolates. Molecular biology is one of the current tools used to differentiate between these isolates. Polymerase Chain Reaction (PCR)-based randomly amplified polymorphic DNA (RAPD) analysis has been used successfully in the analysis of DNA relatedness of species of fungi, bacteria, plants and animals. Dendograms which evaluate/assess the likeness between different isolates has also been used (Martinez et al., 2001). Restriction fragment length polymorphism (RFLP) analysis has been applied to a number of studies to detect differences between fungi and to establish relationships between them. Therefore, the scope of this study was to investigate RAPD analysis (with dendograms) and detection of RFLPs by hybridization as molecular methods that can distinctly differentiate or characterize the aflatoxigenic and non-aflatoxigenic Aspergillus isolates.Item Chemoprotective action of natural products on cultured human epithelial cells exposed to aflatoxin B1(2005) Reddy, Lalini; Odhav, BhartiPrevious studies indicate that a mutation in the non-oncogenic p53 gene is epidemiologically linked to human HCC (Ozturk, 1991; Chan et al., 2003). Hsu et al. (1991) found this link in Chinese, South African and Asian patients and Hollstein et al. (1993) found the same gene mutation in Taiwanese patients. The incidence of these aberrations is reported to be about 20- 50% in HCC’s (Kishimoto et al., 1997). There is sufficient evidence to indicate that carotenoids in addition to their well known antioxidant properties (Paiva and Russel, 1999), also affect intercellular communication, immune responses, neoplastic transformations and growth control, and cellular levels of enzymes that detoxify carcinogens (Zhang et al., 1991; Brockman et al., 1992; Pryor et al., 2000). To date studies carried out have used the rat (Foote et al., 1970; Gradelet et al., 1998) and the mule duckling model (Cheng et al., 2001) to show the protective effect of these carotenoids against AFB1 exposure. Of the well known carotenoids, lycopene and beta- carotene occur in abundance in fruits and vegetables and are safe for human consumption. Aflatoxin B1 frequently induces mutations of the p53 gene which is linked to HCC. Although there is much evidence from epidemiological studies linking the beneficial aspects of carotenoids to the prevention of cancer, the cellular and molecular mechanisms need to be understood in order to implement large scale intervention strategies to prevent AFB1 induced carcinoma. The use of chemical or dietary interventions to alter the susceptibility of humans to the actions of carcinogens and to block, retard or reverse carcinogenesis is an emerging chemoprotective strategy for disease prevention (Abdulla and Gruber, 2000; Kensler et al., 2003; Bingham and Riboli, 2004). Chemoprotection by natural products involves maintaining cellular integrity, preventing DNA alterations, activation of p53 suppressor protein and apoptosis. The aim of this study was thus to investigate the cellular and molecular mechanisms by which beta-carotene and lycopene may prevent the AFB1-induced toxic changes in human hepatocytes. In order to achieve this aim, the following objectives were set out: i. To optimise an in vitro system for the evaluation of AFB1 damage to cultured hepatocytes. ii. To determine the biochemical protection offered by beta-carotene and lycopene to AFB1-exposed hepatocytes, by measuring the mitochondrial activity, cell viability and ROS levels using appropriate enzyme assays and flow cytometry. iii. To determine the cellular protection offered by beta-carotene and lycopene to AFB1-exposed hepatocytes, by studying the morphological changes at the structural and ultrastructural levels using phase contrast light and electron microscopy respectively. iv. To determine the molecular protection offered by beta-carotene and lycopene to AFB1-exposed hepatocytes, by detecting apoptotic bodies as genomic markers and measuring the levels of p53 protein and AFB1-N7-guanine adducts produced.