Medicinal bulbs form part of the diversified flora in South Africa. The plants are used extensively in South African traditional medicine in the treatment of various ailments. Due to the ever-increasing demand and the unrestricted collection of medicinal plants from the wild, many of these slow growing bulbous plant species are driven into over-exploitation and extinction. The main parts collected for use are the underground bulbs, leading to the destructive harvesting of the whole plant. This form of plant harvesting poses threats to the long term sustainability of these plant resources from their natural habitats. Sustainable harvesting of these plants should be within the limits of their capacity for self-renewal. However, this seldom occurs with the often inconsiderate medicinal plant gatherers. Conservation of these plants is therefore necessary. A strategy that would take into consideration the sustainable harvesting and perhaps simultaneously provide similar medicinal benefits, would be the substitution of bulbs with leaves of the same plant. This study was aimed at evaluating the seasonal pharmacological and phytochemical properties in bulbs/corms and leaves of medicinal bulbs with a view of promoting the substitution of bulbs with leaves in traditional medicinal use. Four medicinal bulbous plants, Tulbaghia violacea, Hypoxis hemerocallidea, Drimia robusta and Merwilla plumbea were evaluated for the pharmacological and phytochemical properties in their bulbs/corms and leaves in spring, summer, autumn and winter seasons, with a view of promoting the use of leaves as a conservation strategy. Dried plant materials were sequentially extracted with petroleum ether (PE), dichloromethane (DCM), 80% ethanol (EtOH) and water in each season. The extracts were tested for activities against Gram-positive (Bacillus subtilis and Staphylococcus aureus), Gram-negative (Escherichia coli and Klebsiella pneumoniae) bacteria and the fungus Candida albicans using the in vitro microdilution assays to obtain minimum inhibitory concentrations (MIC) and minimum fungicidal concentrations (MFC). The four plant species were also evaluated for their ability to inhibit cyclooxygenase (COX-1 and COX-2) enzymes. Spectrophotometric methods were used to evaluate saponin and phenolic contents of samples from the four plant species in each season. Antibacterial activity was fairly comparable between bulbs/corms and leaves of H. hemerocallidea, T. violacea, and M. plumbea, with at least one extract showing some good activity (MIC < 1 mg/ml) in most of the seasons. Bulb extracts of D. robusta did not show good antibacterial activity while the leaf extracts showed good activity (0.78 mg/ml) against B. subtilis in spring, summer, and autumn and S. aureus (0.78 mg/ml) in autumn. The best antibacterial activity was recorded in winter, with MIC values as low as 0.195 mg/ml from the DCM bulb extracts of T. violacea against K. pneumoniae and S. aureus and PE corm extracts of H. hemerocallidea (0.195 mg/ml) against B. subtilis. Good antibacterial activity from water extracts were only recorded from corm extracts of H. hemerocallidea in summer, autumn and winter, H. hemerocallidea leaf extracts in autumn and winter, and M. plumbea bulb extracts in autumn. The leaf extracts of all the screened plant species demonstrated good fungicidal activity in autumn, with H. hemerocallidea corm water extracts recording an MFC value as low as 0.39 mg/ml. The leaf extracts of H. hemerocallidea (water), D. robusta (DCM) and M. plumbea (DCM) had good MFC values of 0.78 mg/ml each, in spring. The DCM leaf extracts of T. violacea also showed good fungicidal activity (0.78 mg/ml) in summer, while corm water extracts of H. hemerocallidea had an MFC value of 0.39 mg/ml in winter. There were no fungicidal activities recorded from all the bulb extracts in all the seasons. All the PE and DCM extracts in all the tested plant samples recorded between moderate (40-70%) and high (> 70%) COX-1 and COX-2 inhibition levels across all seasons. The EtOH corm extracts of H. hemerocallidea also demonstrated moderate to high inhibitory activity against COX-1 enzyme across all seasons. Bulb and leaf extracts of T. violacea showed selective inhibitory activity for COX-2 enzyme in all the seasons. The highest COX inhibitory levels were recorded in COX-2 from the PE leaf (spring) and bulb (autumn) extracts of T. violacea, with both recording 100% inhibitory activity. Phytochemical analysis revealed higher total phenolic compounds in bulbs/corms and leaves of all the analysed plant species, to be either higher in spring or winter. Plant material collected in autumn had the least levels of total phenolics. An almost similar trend to that of total phenolics was observed for flavonoids, gallotannins and condensed tannins in most plant samples, with higher levels either in spring or winter. Total saponins were consistently higher in winter than in the other seasons in all the screened plant species. There were in some cases, relationships between the peaks in the levels of some phytochemical compounds and the observed levels of bioactivity in different assays. The results obtained from this study demonstrate that the leaves of the screened plant species may substitute or complement bulbs in the treatment of certain ailments in traditional medicine. Thus, plant part substitution can be sustainably utilised in the conservation of these plant species while retaining the same medicinal benefits. / Thesis (M.Sc.)-University of KwaZulu-Natal, Pietermaritzburg, 2010.

The use of traditional medicine is a popular practice in South Africa especially among rural dwellers due to several reasons such as availability of natural products, cultural beliefs, preference of natural products to synthetically derived drugs and the high cost of modern drugs. Traditional healers in South Africa play key roles in administering treatment for all sorts of ailments using plants. The aim of this study was to evaluate the efficacy of seven selected medicinal plants that are used in traditional medicine to treat stomach-related ailments for their pharmacological and phytochemical properties. Plant material was extracted sequentially with ethyl acetate (EtOAc), ethanol (EtOH) and water. The extracts were evaluated for their antimicrobial activities using the microdilution technique against two Gram-positive (Enterococcus faecalis ATCC 19433 and Staphylococcus aureus ATCC 12600) bacteria and a Gram-negative (Escherichia coli ATCC 11775) bacterium. A modified microdilution technique was used to screen for antifungal activity against a yeast-like fungus (Candida albicans ATCC 10231). Only the EtOAc extract of Tetradenia riparia demonstrated good antibacterial activity against the Gram-negative E. coli, all the other extracts that were active only showed good antibacterial activity against the two Gram-positive (E. faecalis and S. aureus) bacteria with MIC values <1 mg/ml. None of the extracts that exhibited good inhibitory activity showed corresponding bactericidal activity against the bacterial test strains, suggesting that the observed activity were all inhibitory. Good antifungal activity with an MIC value <1 mg/ml was observed in only 5 extracts, and none of the extracts exhibited corresponding fungicidal activity. The in vitro colorimetric assay for anthelmintic activity against Caenorhabditis elegans revealed that almost all the extracts possessed moderate to high anthelmintic properties. The EtOAc extract of T. riparia had the best activity at MLC value of 0.004 mg/ml. The anti-inflammatory activity of the plant extracts was tested using the cyclooxygenase assays to determine their inhibitory potential against COX-1 and COX-2 enzymes. All the EtOAc extracts demonstrated both COX-1 and COX-2 inhibitory activity in the range of 50.7 ± 2.4 to 99.5 ± 0.5%. Apart from the EtOH extracts of C. multicava that showed high inhibitory activity against both COX-1 and COX-2, all the other EtOH extracts were COX-2 selective. Aqueous extracts exhibited poor inhibitory activity against both COX-1 and COX-2 enzymes with the exception of T. riparia and Coddia rudis that showed good inhibitory activity (69.1 ± 0.9 and 92.65 ± 0.7%) against COX-1 and COX-2 respectively. The standard plate incorporation assay for the Ames test was carried out to determine the potential genotoxic effects of the plant extracts and this revealed that all the extracts were non-mutagenic towards Salmonella typhimurium tester strains TA98, TA100 and TA1537 without metabolic activation. However, further studies incorporating metabolizing enzymes are needed to confirm the safe use of the studied plants. Phytochemical analysis revealed relatively high amounts of total phenolics, gallotannins and flavonoids in all the evaluated plants. Total and steroidal saponins were detected in only two plant samples, Canthium spinosum and Cassinopsis ilicifolia (bark). These findings present useful information on the types of bioactive compounds that could be responsible for the pharmacological activities observed among some of the plant extracts. The results obtained in this study showed different levels of pharmacological activities among all the evaluated medicinal plants which provide scientific validation for their use in traditional medicine as antimicrobial agents. Phytochemical analysis provides valuable information for further study that will be aimed at isolation and identification of the bioactive principles in the evaluated plant species. / Thesis (M.Sc.)-University of KwaZulu-Natal, Pietermaritzburg, 2011.

Woo Ka-yan. / "September 2003." / Thesis (Ph.D.)--Chinese University of Hong Kong, 2003. / Includes bibliographical references. / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Electronic reproduction. Ann Arbor, MI : ProQuest Information and Learning Company, [200-] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Mode of access: World Wide Web.

Wong, Ying Chun. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2007. / Includes bibliographical references (leaves 73-78). / Abstracts in English and Chinese. / Acknowledgements --- p.I / Abstract --- p.II / 摘要 --- p.IV / List of Tables --- p.VI / List of Figures --- p.VII / List of Abbreviations --- p.VIII / Chapter Chapter 1 --- Introduction --- p.3 / Chapter 1.1 --- Chemical Constituents of Rhodiola Genus --- p.5 / Chapter 1.1.1 --- Phenylethyl Derivatives --- p.5 / Chapter 1.1.2 --- Phenylpropanoids --- p.8 / Chapter 1.1.3 --- Phenolic Derivatives --- p.11 / Chapter 1.1.4 --- Flavonoids --- p.14 / Chapter 1.1.4.1 --- Flavone and Flavone Glycosides --- p.15 / Chapter 1.1.4.2 --- Flavonols and Their Glycosides --- p.17 / Chapter 1.1.4.3 --- Flavan-3-ol Derivatives --- p.23 / Chapter 1.1.5 --- Monoterpenoids --- p.26 / Chapter 1.1.6 --- Triterpenes --- p.30 / Chapter 1.1.7 --- Miscellaneous Compounds --- p.32 / Chapter 1.2 --- Biological Activities of Rhodiola Genus --- p.33 / Chapter 1.2.1 --- Anti-oxidative Effect --- p.34 / Chapter 1.2.1.1 --- Protective Effect on Ischemia and Reperfusion --- p.34 / Chapter 1.2.1.2 --- Anti-Aging Effect --- p.35 / Chapter 1.2.2 --- Learning and Memory --- p.36 / Chapter 1.2.3. --- Immune Response --- p.37 / Chapter 1.2.4 --- Anti-cancer Effect --- p.38 / Chapter 1.3 --- Objective --- p.39 / Chapter Chapter 2 --- Experimental --- p.40 / Chapter 2.1 --- General Experimental Procedures --- p.40 / Chapter 2.2 --- Plant Materials --- p.40 / Chapter 2.3 --- Extraction and Isolation --- p.41 / Chapter 2.3.1 --- Isolation and Purification of the Ethyl Acetate (E.A.) Fraction --- p.41 / Chapter 2.3.2 --- Isolation and Purification of the Butanol Fraction --- p.44 / Chapter 2.4 --- Characterization of the Isolated Compounds --- p.46 / Chapter 2.4.1 --- β-Sitosterol (1) --- p.46 / Chapter 2.4.2 --- Tyrosol (2) --- p.46 / Chapter 2.4.3 --- trans-Hydroxycinnamic acid (3) --- p.47 / Chapter 2.4.4 --- Geranyl-β-glucopyranoside (4) --- p.47 / Chapter 2.4.5 --- Neryl-β-glucopyranoside (5) --- p.48 / Chapter 2.4.6 --- Hexyl β-Glucopyranoside (6) --- p.48 / Chapter 2.4.7 --- Gallic Acid (7) --- p.49 / Chapter 2.4.8 --- Epigallocatechin-3-Gallate (8) --- p.49 / Chapter 2.4.9 --- Rhodiolgin (9) --- p.50 / Chapter 2.4.10 --- lsolariciresinol-9-β-Glucopyranoside (10) --- p.51 / Chapter 2.4.11 --- Rhodiooctanoside (11) --- p.52 / Chapter 2.4.12 --- Sacranoside B (12) --- p.52 / Chapter Chapter 3 --- Results and Discussion --- p.53 / Chapter 3.1 --- Structural Determination of the Isolated Compounds --- p.53 / Chapter 3.1.1 --- Identification of β-sitosterol (1) --- p.53 / Chapter 3.1.2 --- Identification of Tyrosol (2) --- p.54 / Chapter 3.1.3 --- Identification of trans-Hydroxycinnamic Acid (3) --- p.55 / Chapter 3.1.4 --- Identification of Geranyl-jS-glucopyranoside (4) --- p.56 / Chapter 3.1.5 --- Identification of Neryl-β-glucopyranoside (5) --- p.58 / Chapter 3.1.6 --- Identification of Hexyl β-Glucopyranoside (6) --- p.59 / Chapter 3.1.7 --- Identification of Gallic Acid (7) --- p.60 / Chapter 3.1.8 --- Identification of (-)-Epigallocatechin 3-Gallate (8) --- p.61 / Chapter 3.1.9 --- Identification of Rhodiolgin (9) --- p.63 / Chapter 3.1.10 --- Identification of lsolariciresinol-9-β-glucopyranoside (10) --- p.65 / Chapter 3.1.11 --- Identification of Rhodiooctanoside (11) --- p.67 / Chapter 3.1.12 --- Identification of Sacranoside B (12) --- p.69 / Chapter Chapter 4 --- Conclusion --- p.70 / References --- p.73

Renal Cell Carcinoma (RCC) has low 5 year survival rate and is resistant to radiation and chemotherapy. Phenethyl Isothiocyanate (PEITC) is a naturally occurring phytochemical that has a variety of anti-cancer properties. Here we explore two anti-cancer properties of PEITC: induction of apoptosis and induction of cell cycle arrest in RCC cells and the underlying mechanisms. We used two human RCC cell lines Caki-1 and Caki-2. Survival and cell proliferation was assayed using Calcein AM. Annexin V staining was used to measure apoptosis. Caspase-3/7 induction was measured using a fluorescent substrate. Cell cycle was studied using Propidium Iodide staining. DNA damage was determined using phospho [gamma]-H2AX antibody. Protein expression and phosphorylation was determined using immunoblotting. PEITC significantly reduced survival of Caki-1 and Caki-2 cells and inhibited their proliferation as determined by Calcein AM. 15 and 20 [mu]M PEITC induced apoptosis in both cell lines and induced caspase-3/7 activity. Western blot analysis revealed caspase-8, caspase-9 and Bid cleavage as well as upregulation of the death receptors Fas and DR5. Lower doses (up to 10 [mu]M) arrested Caki-1 cells in G2/M phase, and this was associated with increased p38 and MK2 (Thr334) phosphorylation. The p38 inhibitor SB203850 inhibited this G2 arrest induced by PEITC. 15 and 20 [mu]M PEITC treatment resulted in increased [gamma]-H2AX phosphorylation suggesting DNA damage, but this was completely blocked by caspase inhibitor. In summary, our study shows that PEITC induces apoptosis in Caki-1 and Caki-2 cells by upregulating Fas and DR5 and activating the downstream apoptosis cascade. PEITC does not cause direct DNA damage to the cells; the observed DNA damage is a result of the apoptotic process and is blocked by caspase inhibitor. PEITC induces G2/M arrest in Caki-1 cells and the mechanism involves p38 phosphorylation which activates MK2. Inducing cell cycle arrest and apoptosis may play an important role in the anti-cancer properties of PEITC. Fully understanding the mechanism by which PEITC induces apoptosis and cell cycle arrest in RCC cells may lead to development of novel chemotherapeutic drugs against RCC. / text

Sclerocarya birrea (marula, Anacardiaceae) is a highly-valued indigenous tree in most parts of sub-Saharan Africa because of its medicinal and nutritional properties. The marula tree is adapted to the semi-arid conditions that characterise most parts of sub-Saharan Africa and renders them unsuitable for conventional crop agriculture. The unique nutritional properties of marula and its high tolerance to dry conditions provide opportunities for its development into a plantation crop. On the other hand, the demand for marula plant parts, mainly the bark and roots as medicinal remedies, poses a great threat to wild populations. In the long term, the growing demand of marula products in the food, pharmaceutical and cosmetic industries will not be sustainable from wild populations alone. Plant tissue culture technologies can be useful for in vitro manipulation and mass propagation of the plant in the process of domestication and conservation. The aims of the project were to determine the optimum conditions for seed germination, in vitro propagation and plant regeneration, and to evaluate the potential bioactivity of secondary metabolites from its renewable plant parts as an alternative option in the conservation of S. birrea. An ex vitro seed germination study indicated that after-ripening and cold stratification are critical factors. Cold stratification (5 °C) of marula nuts for 14 days improved germination (65%) as compared to non-stratified nuts (32%). Direct shoot organogenesis was achieved from leaf explants through the induction of nodular meristemoids on Murashige and Skoog (MS) (1962) medium and woody plant medium (WPM) supplemented with 6-benzyladenine (BA) in combination with naphthalene acetic acid (NAA), indole-3-butryric acid (IBA) and indole-3-acetic acid (IAA). Induction of nodular meristemoids from 86% of the leaf cultures was achieved on a MS medium with 4.0 ìM BA and 1.0 ìM NAA. High levels (78–100%) of induction were also achieved on WPM with different concentrations of BA (1.0–4.0 ìM) and IBA (1.0–4.0 ìM). The highest conversion of nodular meristemoids into shoots on MS initiation medium was only 22% for 4.0 ìM BA and 1.0 ìM NAA. This was improved to 62% when nodular clusters were cultured in MS liquid medium. Histological studies revealed high numbers of unipolar meristematic buds developing from globular nodules. These embryo-like structures have in the past been mistaken for true somatic embryos. The initiation of high numbers of nodular meristemoids per explant provides potential for automated large-scale clonal propagation in bioreactors, in vitro phytochemical production and the development of synthetic seed technology, similar to somatic embryogenesis. Plant regeneration through nodule culture has potential for application in mass micropropagation and plant breeding of S. birrea. Adventitious shoot and root induction are important phases in micropropagation. Plant growth regulators play an important role in these developmental processes, and the type and concentration used have major influences on the eventual organogenic pathway. Three auxins (IAA, IBA and NAA) and four aromatic cytokinins (6-benzyladenine, meta-topolin, meta-topolin riboside, and meta-methoxytopolin riboside) were evaluated for their potential to induce adventitious shoot and root formation in S. birrea shoots, hypocotyls and epicotyls. Among the evaluated cytokinins, the highest adventitious shoot induction (62%) was achieved on MS medium supplemented with meta-topolin (8.0 ìM). The lowest adventitious shoot induction (2.5%) was obtained on MS basal medium containing 2.0 ìM meta-methoxytopolin riboside. The highest adventitious shoot induction for hypocotyls was 55% on MS medium supplemented with 8.0 ìM meta-topolin. For the tested auxins, IBA induced adventitious rooting in 91% of shoots at a concentration of 4.0 ìM after 8 weeks in culture. However, the in vitro rooted plants only survived for two weeks when transferred ex vitro. A temperature of 25 °C and 16-h photoperiod were optimum for adventitious root induction. Stomatal density (number per mm2) on the abaxial leaf surfaces was higher for the 16-h photoperiod treatment (206.6 ± 15.28) compared to that for a 24-h photoperiod (134.6 ± 12.98). Normal mature stomata with kidney-shaped guard cells and an outer ledge over the stomatal pore were observed for in vitro plants growing under a 16-h photoperiod. Total phenolic content, proanthocyanidins, gallotannins, flavonoids, and antioxidant activities of S. birrea methanolic extracts were evaluated using in vitro bioassays. Methanolic extracts of the young stem bark and leaves contained high levels of these phytochemicals. Sclerocarya birrea young stem extracts contained the highest levels of total phenolics (14.15 ± 0.03 mg GAE g-1), flavonoids (1219.39 ± 16.62 ìg CE g-1) and gallotannins (246.12 ± 3.76 ìg GAE g-1). Sclerocarya birrea leaf extracts had the highest concentration of proanthocyanidins (1.25%). The EC50 values of the extracts in the DPPH free radical scavenging assay ranged from 5.028 to 6.921 ìg ml-1, compared to ascorbic acid (6.868 ìg ml-1). A dose-dependent linear curve was obtained for all extracts in the ferric-reducing power assay. All the extracts exhibited high antioxidant activity comparable to butylated hydroxytoluene based on the rate of â-carotene bleaching (89.6 to 93.9%). Sclerocarya birrea provides a source of secondary metabolites which have potent antioxidant properties and may be beneficial to the health of consumers. Sclerocarya birrea young stem and leaf ethanolic extracts exhibited high bioactivity (MIC < 1.0 mg ml-1) against both Gram-positive (Bacillus subtilis and Staphylococcus aureus) and Gram-negative (Escherichia coli and Klebsiella pneumoniae) bacteria. The highest activity (MIC = 0.098 mg ml-1 and total activity = 1609.1 ml g-1) was recorded for young stem extracts against B. subtilis. The highest activity (MIC = 1.56 mg ml-1 and MFC = 1.56 mg ml-1) in the antifungal assay against Candida albicans was observed for young stem ethanolic extracts. Sclerocarya birrea extracts had moderate acetylcholinesterase (AChE) inhibition activity. The dichloromethane (DCM) and methanol (MeOH) fractions exhibited dose-dependent acetylcholinesterase inhibitory activity. The highest AChE inhibitory activities were from leaves (DCM fraction, IC50 = 0.1053 mg ml-1) and young stems (MeOH fraction, IC50 = 0.478 mg ml-1). High inhibitory activity against cyclooxygenase (COX-1 and COX-2) enzymes was observed. All extracts and fractions showed high COX-1 enzyme inhibition (90.7-100%). Petroleum ether (PE) and dichloromethane fractions also exhibited high inhibition against COX-2 enzyme (77.7-92.6%). The pharmacological activities observed suggest that S. birrea renewable plant parts (leaves and young stems) provide a substantial source of medicinal secondary metabolites. Based on these results, plant part substitution can be a practical conservation strategy for this species. / Thesis (Ph.D.)-University of KwaZulu-Natal, Pietermaritzburg, 2009.

The crisis of newly emerging diseases and the resistance of many pathogens to currently used drugs, coupled with the adverse side-effects of many of these drugs have necessitated the continuous search for new drugs that are potent and efficacious with minimal or no adverse side-effects. The plant kingdom is known to contain many novel biologically active compounds, many of which could potentially have a higher medicinal value when compared to some of the current medications. Indeed, the use of plants in traditional medicine, especially in African communities, is gaining more importance due to their affordability and accessibility as well as their effectiveness. Exponential population growth rates in many developing countries has resulted in heavy exploitation of our plant resources for their medicinal values. In addition, plant habitat destruction arising from human developmental activities has contributed to the fragmentation or loss of many plant populations. Owing to these factors, many plant species with horticultural and/or medicinal potential have become either extinct or are threatened with extinction. These threatened species cut across different taxonomic categories including shrubs, trees and succulents. Without the application of effective conservation strategies, the medicinal and/or horticultural potential of such threatened species may be totally lost with time. The extinction of such species could lead to the loss of potential therapeutic compounds and/or genes capable of being exploited in the biosynthesis of new potent pharmaceutical compounds. / Thesis (Ph.D.)-University of KwaZulu-Natal, Pietermaritzburg, 2009.

Among all the major infectious human diseases, gastro-intestinal infections caused by microbial pathogens are a major cause of morbidity and infant death in developing countries, largely due to inadequate sewage disposal and contaminated water. Traditional health practitioners in South Africa play a crucial role in providing health care to the majority of the population. Many plants are locally used by South African traditional healers to treat microbial infections related to gastro-intestinal tracts. Ethnopharmacological and ethnobotanical studies using traditional knowledge as a selection strategy has given priority to certain plants for isolation and identification of plant novel bioactive compounds. Pharmacological and phytochemical studies of the investigated twelve medicinal plant species (from 10 families) extensively used as antimicrobials against gastro-intestinal infections was necessary to validate the use of the plants. Furthermore, to provide sufficient preliminary information for the isolation and identification of active compounds that are present in the investigated plants. Plant parts were sequentially extracted using petroleum ether (PE), dichloromethane (DCM) and 70% ethanol (EtOH). Cold water and boiled (decoction) extracts of the plant materials were prepared non- sequentially. Among the extracts, EtOH yielded the highest amount of plant substances. A total number of 85 extracts were evaluated for antibacterial activity, 80 for antifungal activity, 64 for anti-inflammatory activity, and 27 biologically active extracts were tested for genotoxicity. The microdilution method was used to determine the minimum inhibitory concentration values in the antibacterial assay against two Gram-negative bacteria (Escherichia coli ATCC 11775 and Klebsiella pneumoniae ATCC 13883) and two Gram-positive bacteria (Bacillus subtilis ATCC 6051 and Staphylococcus aureus ATCC 12600). A modified microdilution method was used to determine the minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) values in the antifungal assay against Candida albicans. Cyclooxygenase assay was used to evaluate the anti-inflammatory activity of the extracts against cyclooxygenase-1 and -2 (COX-1 and COX-2) enzymes. The plant extracts were screened first at a concentration of 250 ƒÊg/ml per test sample, and then further screened at concentrations of 125 and 62.5 ƒÊg/ml for extracts that inhibited the COX-2 enzyme. The Ames test was used to test for genotoxicity in extracts that showed interesting pharmacological activities using Salmonella typhimurium strain TA98. Among the screened extracts, 25 extracts showed good antibacterial activity with MIC values . 1.0 mg/ml. Dichloromethane extracts exhibited the greatest antibacterial activity, and Gram-positive bacteria were most susceptible. The best antibacterial activity was exhibited by Becium obovatum leaf EtOH extracts with an MIC value of 0.074 mg/ml. A broad spectrum antibacterial activity was observed by leaf extracts of Cucumis hirsutus (PE), Haworthia limifolia (PE), Protea simplex (PE and DCM) and Dissotis princeps (EtOH) against both Gram-negative and Gram-positive bacteria. No interesting antibacterial activity was exhibited by water extracts with the exception of Dissotis princeps water extract with a good antibacterial activity against Gram-positive and Gram-negative bacteria. In the antifungal assay, 6 extracts showed interesting antifungal activity. Protea simplex leaf PE extract showed the best fungicidal activity with an MFC value of 0.014 mg/ml. The best overall antifungal activity was observed in plant EtOH extracts. Some extracts from Agapanthus campanulatus (leaves and roots), Dissotis princeps (leaves), Gladiolus dalenii (corms) and Protea simplex (leaves) showed good activity against Candida albicans. Twenty one extracts inhibited the COX-1 enzyme, while fifteen extracts inhibited the COX-2 enzyme at the lowest screening concentration of 62.5 ƒÊg/ml. The highest COX-1 inhibition at a concentration of 62.5 ƒÊg/ml was exhibited by Diospyros lycioides leaf PE extract (89.1%) while Agapanthus campanulatus root DCM extract showed the highest COX-2 inhibitory activity (83.7%) at the same concentration. In the Ames test, no genotoxicity was observed in any of the extracts, however more tests need to be done to confirm these results. Thin layer chromatograms of the organic solvent plant extracts were developed. The fingerprints of the plant extracts showed colours of bands at different Rf values when viewed under UV254 and UV366 suggesting that the investigated plant species contained different compounds in the extracts. In the quest to understand the source of the plants pharmacological activities, total phenolic compounds including condensed tannins, gallotannins and flavonoids were quantitatively investigated in terms of their amounts in the aqueous methanol extracts of the plants materials using spectrophotometric methods. Alkaloids and saponins were qualitatively determined. The amounts of total phenolics were determined by the Folin Ciocalteu assay, condensed tannins were determined by the butanol-HCl assay, while rhodanine and vanillin assays were used to determine the amounts of gallotannins and flavonoids respectively. Dragendorff reagent was used to detect alkaloids in the plant extracts on thin layer chromatographic plates, while the froth test was employed to detect saponins. Secondary metabolites varied with plant parts and species with Cyperus textilis (leaf) having the highest amounts of total phenolics, condensed tannins and flavonoids. The highest amount of gallotannins was detected in Protea simplex leaf extracts. All the investigated plant materials with the exception of Haworthia limifolia leaf, Protea simplex leaf, Antidesma venosum leaf and Dissotis princeps leaf tested positively to saponins. Alkaloids were detected in Haworthia limifolia leaf (PE and EtOH), Cucumis hirsutus leaf (EtOH), Becium obovatum root (DCM), Protea simplex root and bark (EtOH), Agapanthus campanulatus root (DCM) and leaf (EtOH), Cyperus textilis root (DCM), Vernonia natalensis leaf (PE), Antidesma venosum leaf (PE), Diospyros lycioides leaf (PE) and Dissotis princeps leaf (DCM) extracts. The results obtained from the investigation of the pharmacology and phytochemistry of the plant species used to treat microbial infections related to gastro-intestinal tracts, provide sufficient preliminary information to validate the use of some of the plants in traditional medicine. The information provided might be considered sufficient for further studies aimed at isolating and identifying the active compounds in the plant species, and evaluating possible synergism amongst the isolated compounds. / Thesis (M.Sc)-University of KwaZulu-Natal, Pietermaritzburg, 2009.

Traditional medicine in South Africa is part of the culture of the people and has been in existence for a long-time. Although animal components form part of the ingredients used, plant material constitutes the major component. South Africa is endowed with vast resources of medicinal and aromatic plants which have been employed for treatment against various diseases for decades. A large number of South Africans still depend on traditional medicine for their healthcare needs due to its affordability, accessibility and cultural importance. Helminth infections are among the variety of diseases treated by traditional healers. These infections are regarded as neglected tropical diseases (NTDs) due to their high prevalence among the economically disadvantaged living in rural areas in different regions of the world. / Thesis (M.Sc.)-University of KwaZulu-Natal, Pietermaritzburg, 2009.

Certain crop plants are susceptible to pathogens or unable to develop efficient microbial symbioses. These crops adversely impact soil biological quality with consequences on plant health and productivity of cropping systems. Chickpea is a rotational pulse crop with two types: kabuli and desi, and several cultivars. Cultivation of chickpea has inconsistent effects on soil microbial communities and subsequent wheat crops. I conducted field studies and used high throughput molecular analyses to explore the variations among chickpeas to identify cultivars developing fungal communities that are conducive to plant health and productivity. I also carried out greenhouse studies and used biochemical analyses to investigate the response of chickpea cultivars to arbuscular mycorrhizal (AM) fungi and non-AM fungal endophytes and identify the influence of root and root metabolites on the endophytic and pathogenic fungi. Cultivars and types of chickpeas and environmental conditions promoted different fungal communities in the root endosphere. Funneliformis and Claroideoglomus were the dominant AM fungal genera and Fusarium and Alternaria were the dominant non-AM fungal genera in the roots of chickpea. The roots of cultivars CDC Corrine, CDC Cory and CDC Anna hosted the most diverse fungal communities in contrast to CDC Alma and CDC Xena roots which hosted the least diverse communities. Plant response to AM and non-AM fungal endophytes varied with genotype and type of chickpea. The root symbiosis effectively promoted plant growth in CDC Cory, CDC Anna and CDC Frontier and stimulated nitrogen fixation in CDC Corrine. Cultivars of chickpea responded differently to dual inoculation of the AM and non-AM fungal endophytes. Co-inoculation with AM and non-AM fungal endophytes had additive effects on CDC Corrine, CDC Anna and CDC Cory but non-AM fungal endophytes reduced the positive effect of AM fungi in Amit and CDC Vanguard. Desi chickpea appeared to form more efficient symbioses with soil fungal resources than kabuli chickpea. Protein(s) up-regulated in the mycorrhizal roots of the desi chickpea CDC Anna suppressed the growth of the fungal endophytes Trichoderma harzianum and Geomyces vinaceus and of the pathogens Fusarium oxysporum and Rhizoctonia sp. The formation of AM symbiosis decreased the production of root bioactive metabolites soluble in 25% methanol. Some of the root metabolites stimulated the growth of Trichoderma harzianum and Geomyces vinaceus, and a few inhibited Rhizoctonia sp. and Fusarium oxysporum. A few metabolites with contrasting effects on the different fungal species were detected. The non-protein phytochemicals had selective effects on the endophytes and pathogens whereas the antifungal proteins of mycorrhizal roots were non-selective. Overall the study reveals a "genotype effect" of chickpea on the soil microbiota suggesting the possibility to improve the performance of this crop through the selection of genotypes improving the communities of root associated fungi, by associating and responding to beneficial fungi and repressing the pathogens.