X, 125p / Thesis (M.Sc. (Botany)) --University of Limpopo, Turfloop Campus, 2010 / A study of the role played by muthi shops and street vendors on the trade of indigenous medicinal plants of the Limpopo Province was undertaken in order to develop strategies that will prevent further loss of wild population. This study further investigated the conservation status and in situ availability of targeted medicinal plants, as well as suitable methods to replace wild collections with cultivated ones. Existing environmental legislation was interrogated to ascertain its effectiveness in practice. Nearly 231 medicinal plants were traded at the 16 investigated muthi shops and street vendors, accounting for a calculated 0.96 tonnes of plant material traded annually. Roots were the most preferred item traded. Open access communal lands are the main supply source for muthi markets, coupled with the destructive harvesting methods and involvement of unscrupulous middlemen in collecting medicinal material possesses a serious impact on the survival of medicinal plants. The above factors have already led to a significant decline in the availability of some species in the Limpopo Province. The cultivation of the eight most frequently encountered medicinal species was investigated. Cultivation information provided by indigenous nurseries indicates that medicinal plant species can be cultivated. Efforts to protect species through national and provincial legislation have been ineffective. Environmental laws were totally unknown by all of muthi traders interviewed. Unregulated exploitation of medicinal plants continued regardless of the fact that environmental compliance officers are aware of legislative protection given to specific species and plants in general. Failure to stabilize the status of medicinal plants in Limpopo Province will have not only negative effect on the Province environment, but also on the overall health status of the majority of people living in Limpopo Province. / N/A

Since ancient times, indigenous plants have been used by traditional healers for treating various ailments. Sutherlandia frutescens is one of the most commonly used medicinal plants of southern Africa. This widely distributed plant has been traditionally used to treat cancer and HIV patients; however scientific validation is still in high demand. This research aimed to phytochemically characterise the various extracts prepared and to determine if any chemotypes were present. Subsequent biological characterisation was carried out to preliminary ascertain whether this medicinal plant could have anti-cancer and/or immunemodulating properties and which compounds might be responsible for these actions. Various traditional and organic extracts were prepared. Extracts, fractions and compounds generated were analysed and chemical profiles obtained. Column chromatographic techniques were used to isolate and purify compounds and structure elucidation was carried out using various analytical techniques. Sulforhodamine B and cytometric bead array assays were performed to determine the biological activities of samples generated. / Life and Consumer Sciences / (M. Sc. (Life Sciences)

Text in English / The introduction of the New Growth Path (NGP) and the New Industrial Policy Framework (NIPF) have created an opportunity for the industrialisation of indigenous medicinal and aromatic plants (MAPs Amarula (Sclerocarya birrea), Buchu (Agathosma betulina), and Pelargonium (Pelargonium sidoides) for job creation and economic development in South Africa. The aim is to create a locally sustainable industry which creates jobs as well as improvement of the balance of payments. The National Growth Plan highlights some of the opportunities for industrialization as well as creation of decent jobs. There is increasing pressure on the harvesting of wild indigenous medicinal and aromatic plants (MAPS). The establishment of a local pharmaceutical and nutraceutical industry for processing MAPS remains a challenge in South Africa and many low income countries. The objective of this study was to assess the challenges and their impact looking at particular species, Amarula, Buchu and Pelargonium by using structured focus group interviews with users in communities and businesses in KwaZulu Natal Province, Eastern Cape and the Western Cape. More data/information was also collected from key stakeholder companies including desk-top reviews. Data/information was analysed using the SPSS computer programme and the Atlas software to summarize data and the results. Graphs and tables were used where necessary. The qualitative research reported on some qualitative assessments and challenges facing the establishment of medicinal and aromatic plants in South Africa. Comparative findings from BRICS were also reported. The results of the study indicated that there is continuous exploitation of several medicinal plant species, for plant trade, from the wild and substantial loss of their habitat. The research identifies a number of challenges such as: poor standards for raw materials, lack of research and development linking industry driven revenues, demand exceeding supply, regulatory risk, community risk, lack of understanding of judicious agronomic practices, ethical challenges, liquidity risk as well as political instability fail to attract foreign direct investments. The research concludes that for the establishment of a viable local pharmaceutical industry, iii beverage industry and nutraceutical industry with sustainable harvesting, there are serious hegemonic challenges which will require multi-stakeholder analysis. However, for the South African (SA) industry to be competitive, detailed feasibility studies need to be done comparing SA with other BRICS countries. There is also a need for the Research and Development to be strengthened. It must also be linked into industrial development in the industry. Furthermore there is a need to initiate the incentivizing of the pharmaceutical manufacturing to lower the cost of setting up the businesses in the industry and lessen the need to harvest from South African forests. Furthermore there is a need for programmes of production so that key species are given opportunity for cultivation and the attention they deserve to preserve these depleting resources. / Agriculture and  Animal Health / M.Sc. (Agriculture)

Text in English / The introduction of the New Growth Path (NGP) and the New Industrial Policy Framework (NIPF) have created an opportunity for the industrialisation of indigenous medicinal and aromatic plants (MAPs Amarula (Sclerocarya birrea), Buchu (Agathosma betulina), and Pelargonium (Pelargonium sidoides) for job creation and economic development in South Africa. The aim is to create a locally sustainable industry which creates jobs as well as improvement of the balance of payments. The National Growth Plan highlights some of the opportunities for industrialization as well as creation of decent jobs. There is increasing pressure on the harvesting of wild indigenous medicinal and aromatic plants (MAPS). The establishment of a local pharmaceutical and nutraceutical industry for processing MAPS remains a challenge in South Africa and many low income countries. The objective of this study was to assess the challenges and their impact looking at particular species, Amarula, Buchu and Pelargonium by using structured focus group interviews with users in communities and businesses in KwaZulu Natal Province, Eastern Cape and the Western Cape. More data/information was also collected from key stakeholder companies including desk-top reviews. Data/information was analysed using the SPSS computer programme and the Atlas software to summarize data and the results. Graphs and tables were used where necessary. The qualitative research reported on some qualitative assessments and challenges facing the establishment of medicinal and aromatic plants in South Africa. Comparative findings from BRICS were also reported. The results of the study indicated that there is continuous exploitation of several medicinal plant species, for plant trade, from the wild and substantial loss of their habitat. The research identifies a number of challenges such as: poor standards for raw materials, lack of research and development linking industry driven revenues, demand exceeding supply, regulatory risk, community risk, lack of understanding of judicious agronomic practices, ethical challenges, liquidity risk as well as political instability fail to attract foreign direct investments. The research concludes that for the establishment of a viable local pharmaceutical industry, iii beverage industry and nutraceutical industry with sustainable harvesting, there are serious hegemonic challenges which will require multi-stakeholder analysis. However, for the South African (SA) industry to be competitive, detailed feasibility studies need to be done comparing SA with other BRICS countries. There is also a need for the Research and Development to be strengthened. It must also be linked into industrial development in the industry. Furthermore there is a need to initiate the incentivizing of the pharmaceutical manufacturing to lower the cost of setting up the businesses in the industry and lessen the need to harvest from South African forests. Furthermore there is a need for programmes of production so that key species are given opportunity for cultivation and the attention they deserve to preserve these depleting resources. / Agriculture and  Animal Health / M. Sc. (Agriculture)

Since ancient times, indigenous plants have been used by traditional healers for treating various ailments. Sutherlandia frutescens is one of the most commonly used medicinal plants of southern Africa. This widely distributed plant has been traditionally used to treat cancer and HIV patients; however scientific validation is still in high demand. This research aimed to phytochemically characterise the various extracts prepared and to determine if any chemotypes were present. Subsequent biological characterisation was carried out to preliminary ascertain whether this medicinal plant could have anti-cancer and/or immunemodulating properties and which compounds might be responsible for these actions. Various traditional and organic extracts were prepared. Extracts, fractions and compounds generated were analysed and chemical profiles obtained. Column chromatographic techniques were used to isolate and purify compounds and structure elucidation was carried out using various analytical techniques. Sulforhodamine B and cytometric bead array assays were performed to determine the biological activities of samples generated. / Life and Consumer Sciences / (M. Sc. (Life Sciences)

PhD (Chemistry) / Department of Chemistry / Terminalia sericea Burch. ex. DC (Combretaceae) is one of the 50 most popular medicinal plants in Africa. The fruit, leaves, stems and roots are commonly used for the treatment of cough, skin infections, diabetes, diarrhoea, venereal diseases and tuberculosis. However, the roots are most commonly used in the preparation of traditional medicines. Pharmacological studies have revealed that the crude root extracts display antibacterial activity against several Gram-positive and Gram-negative bacteria. Anolignan b, termilignan b and arjunic acid are reported to be the major antibacterial constituents present in the roots. Other compounds isolated from the roots include resveratrol-3-rutinoside, sericic acid, sericoside and arjunglucoside I. Authorities worldwide, including the Medicines Control Council of South Africa, have begun to regulate herbal drugs sold in the form of commercial formulations. Quality control of herbal drugs is challenging, since the chemical profiles of the raw materials may vary, depending on the origin of the plant material and the way that it was handled and processed. The chemistry, in turn, impacts on the safety and efficacy of the plant material. To date, there are no available data on parameters that can be used to standardise the quality of T. sericea raw materials. The aim of this study was therefore to provide information on the variation of the chemical constituents that contribute to the biological effects of the roots of T. sericea and also establish its safety. Since the compounds previously isolated from the roots were not commercially available, isolation of the major constituents of the roots was undertaken to obtain analytical standards. A crude dichloromethane:methanol (1:1) extract was initially fractionated using silica gel column chromatography, where after, some of the fractions were further purified using silica gel and Sephadex LH-20 column chromatography. Final purification of the enriched fractions was achieved using preparative high performance liquid chromatography coupled with mass spectrometry (prep-HPLC-MS). The structures of these compounds were subsequently elucidated using one- and two- dimensional nuclear magnetic resonance (NMR) spectroscopy and mass spectrometry and identified as sericic acid (340 g), sericoside (500 g), resveratrol-3-rutinoside (240 mg) and arjunglucoside I (74 mg). The chemical variation within the crude root extracts of samples (n = 42) from ten populations in the Limpopo Province of South Africa, was determined using ultra performance liquid chromatography-mass spectrometry (UPLC-MS). A method was developed for the simultaneous determination of sericic acid, resveratrol-3-rutinoside, sericoside and arjungluicoside I in the extracts using UPLC with photodiode array detection (PDA). The method was validated according to the guidelines of the International Council for Harmonisation (ICH). A regression coefficient (R2) of 0.998 was obtained for sericic acid, resveratrol-3- rutinoside and arjunglucoside I, while the R2 value for sericoside was 0.999, indicating a linear relationship between the concentration and the detector response. Satisfactory limits of detection for sericic acid (25.2 ng/mL), sericoside (11.6 ng/mL), resveratrol-3-rutinoside (23.3 ng/mL) and arjunglucoside I (8.81 ng/mL) were determined. Recoveries of 98 % and 80% were obtained for samples spiked with 12.5 μg/mL and 25 μg/mL of resveratrol-3-rutinoside, respectively, indicating that the method is accurate. The intra- and inter-day variation in resveratrol-3-rutinoside concentration, measured over three days, indicated excellent analytical precision, since all the relative standard deviations were below 0.70 %. The quantitative data revealed that sericic acid (1.59 to 8.45 mg/g), sericoside (2.07 to 20.17 mg/g), resveratrol-3-rutinoside (0.65 to 29.82 mg/g) and arjunglucoside I (0.86 to 8.44 mg/g dry weight) were the major constituents of the root samples, but their concentrations were highly variable. Chemometric analysis of the aligned UPLC-MS data was used to investigate similarities and differences in the chemical profiles of the samples using an untargeted approach. A principal component analysis (PCA) model was constructed and subsequently hierarchical cluster analysis (HCA) indicated the presence of two main groups, which were found to be independent of the populations to which the samples belong. Classes, based on the HCA class identifiers, were subsequently assigned to the samples, and an orthogonal projection to latent structures-discriminant analysis (OPLS-DA) model was then constructed, (R2 cum = 0.996 and Q2 cum = 0.967). The corresponding loadings plot allowed sericic acid, sericoside and resveratrol-3-rutinoside to be identified as biomarkers associated with the first group. Quantitative, rather than qualitative differences were responsible for the observed clustering pattern. Techniques that could be applied in quality control protocols for T. sericea root were investigated. High performance thin layer chromatography (HPTLC) analysis of the root extracts was optimised by testing different developing solvents and visualization reagents. The presence of the sericic acid (Rf = 0.80), sericoside (Rf = 0.49) and resveratrol-3-rutinoside (Rf = 0.36) were clearly visible on the plates. There were visible variations in the concentrations of resveratrol-3-rutinoside in representative samples from the 10 populations, corresponding to the UPLC results. The powdered samples were then analysed by mid-(MIR) infrared spectroscopy. Chemometric analysis of the data revealed no definitive clustering pattern. Partial least squares-discriminant analysis (PLS-DA) calibration models were established from the MIR spectral data combined with the accurate UPLC-values, for the prediction of the sericoside (R2Y = 0.848, Q2 = 0.757, RMSEP = 2.70 mg/g) and resveratrol-3-rutinoside (R2Y = 0.794, Q2 = 0.695, RMSEP = 4.37 mg/g) concentrations in powdered root samples. The antibacterial activities of the root extracts, column fractions and isolated compounds were determined using three Gram-positive and five Gram-negative bacteria, all selected due to their ability to cause intestinal and skin disorders. Extracts and fractions containing high concentrations of sericic acid exhibited the highest activities against Klebsiella pneumoniae (ATCC 13883), Pseudomonas aeruginosa (ATCC 27858), Salmonella typhimurium (ATCC 14028), Staphylococcus aureus (ATCC 25923), Staphylococcus epidermidis (ATCC 12223) and Shigella sonnei (ATCC 9292). The pure compound (sericic acid) was highly active against S. sonnei (MIC 0.078 μg/mL), a Gram- negative bacterium. There were no variations in the activity of the crude extracts against B. cereus and P. aeruginosa, while the MIC values obtained against S. typhi were variable and ranged from 0.25 to 1.0 mg/mL. Sericoside and resveratrol-3-rutinoside did not display any activity. The anti-oxidant activities were evaluated using DPPH (2,2-diphenyl-1-picrylhydrazyl) and reducing power assays. The anti-oxidant assays revealed that resveratrol-3- rutinoside exhibited lower activity (DPPH = 186 μg/mL; RP = 184 μg/mL) compared to the crude extract (DPPH = 22.3 μg/mL; RP = 24.4 μg/mL) and ascorbic acid (DPPH = 11.3 μg/mL, RP = 145 μg/mL). Sericic acid and sericoside did not display any anti- oxidant activities. The variation in the anti-oxidant activities (4.58 to 26.0 μg/mL) of the samples from different populations was an indication of chemical variability. A toxicity study of the raw powdered plant material was conducted using vervet monkeys (Chlorocebus pygerythrus). Biochemical analysis (liver function tests, kidney function tests and hematology), physical and physiological examinations were conducted. The subjects were fed a normal diet supplemented with T. sericea root powder (2.14 g/kg per day) for 120 days, where after the diet was returned to normal (washout) for another 30 days. The treatment groups presented with elevated serum enzymes at Week 4, followed by the reduction of the elevated serum enzymes levels at Week 12. These results indicate short-term hepatotoxic effects, followed by hepatoprotective activity. Reduction of the serum glucose at Week 4 suggests hypoglycemic potential. However, elevated serum creatinine levels indicated possible nephrotoxicity. In conclusion, this study has indicated the variability in the chemical constituents of the roots of T. sericea, which affects the antibacterial and anti-oxidant activities. Sericic acid, resveratrol-3-rutinoside, and sericoside were, for the first time, identified as biomarkers that can be used for the quality control of raw root material to be used in herbal products. Sericic acid was also found to be the main antibacterial constituent of the roots. The hepatoprotective, nephrotoxic and hematotoxic effects observed in monkeys to which the root powder had been administered is cause for concern. / NRF