The rising costs of prescription drugs in the maintenance of personal health and wellbeing have increased the interest in medicinal plants. The World Health Organization estimates that 65 percent-80 percent of the world’s population use traditional medicine as their primary form of health care. In this project the focus has been on the use of Leonotis leonurus extracts as a traditional medicine. The major chemical constituent of this plant is marrubiin, which is a diterpenoid labdane lactone formed from a precursor called premarrubiin. Aqueous and acetone extract (AL and OL extract, respectively) of this plant has been found to have an antithrombotic effect, with IC50 values of 3mg/ml and 6mg/ml, respectively. The extracts also have an effect on fibrinolysis, where the lysis time was decreased by more than 50 percent by the organic extract and standard marrubiin. In whole blood ADP-induced platelet aggregation, the organic extract inhibited aggregation by 68 percent at a final concentration of 138μg/ml (equivalent to 7.2μg/ml marrubiin). Marrubiin has also been screened for antithrombotic/anticoagulant activity; no antithrombotic activity has been observed but it increased the rate of fibrinolysis, by decreasing lysis time by 64 percent and also decreasing fibrin formation. From these findings it can be concluded that marrubiin has a fibrinolytic effect and antiplatelet aggregation effect. In the diabetic studies, in hyperglycemic condition, the OL (10μg/ml) extract and standard marrubiin significantly increased insulin secretion by 200 percent (2-fold) and 400 percent (4-fold), respectively, with respect to the control. The OL extract and standard marrubiin stimulated the release of insulin, the stimulatory index was significantly increased by 450 percent (4.5-fold) and 500 percent (5-fold), respectively, with respect to the control. In the apoptotic studies, in the normoglycemic and hyperglycemic conditions, the OL extract decreased the occurrence of apoptosis, in a dose-dependent manner, with the lower concentrations inducing apoptosis significantly higher than the relevant controls. Standard marrubiin did not have an effect on apoptosis in hyperglycemic condition, but it decreased the occurrence of apoptosis by 200 percent (2-fold) under normoglycemic conditions. The OL extract increased proliferation by 148 percent (1.48- fold) and 155 percent (1.55-fold) in normoglycemic and hyperglycemic conditions, respectively. The same effect was observed for standard marrubiin, where, proliferation was increased by 180 percent (1.8-fold) and 200 percent (2.0-fold) in normoglycemic and hyperglycemic conditions, respectively. RT-PCR displayed that standard marrubiin inhibited the expression of insulin by 50 percent under normoglycemic conditions.

It has been estimated that between 3000 and 4000 plant species are used for their medicinal properties throughout South Africa, with approximately 27 million South Africans making use of traditional medicines. Of this 27 million, 3 million South Africans rely on traditional medicine as their primary source of health care. Of the 250 000 to 500 000 known plant species, very few have been investigated for their pharmacological qualities, and compounds of significant medicinal value may still remain undiscovered in many plant species. The aims of this study included investigating the antimicrobial properties of Geranium incanum and Artemisia afra, both plants traditionally used for their medicinal properties, and comparing the antimicrobial activity of the latter to that of Artemisia absinthium, as well as investigating the anticancer properties of G. incanum and A. afra, and comparing the anticancer activity of the latter to that of A. absinthium. Infusions, aqueous-, methanol- and acetone extracts of the three plants were prepared and used for anticancer and antimicrobial screening. Plant specimens used to prepare extracts for antimicrobial activity were collected and extracted over three seasons, while extracts used for anticancer screening were prepared from plants collected during the summer only. Considerable variation existed in the percentage crude extract yields obtained when different extractants were used, while the season in which the plants were harvested and extracted also appeared to play a significant role in the amount of extract obtained. The plant extracts were screened for antimicrobial activity against various strains of Candida albicans, Escherichia coli, Pseudomonas aeruginosa, Enterococcus faecalis, Staphylococcus aureus and Bacillus cereus, using an agar dilution method. G. incanum and A. afra possessed activity for C. albicans, while all three plants showed activity for S. aureus and B. cereus. Activity was largely dependent on the extraction method used. iii The 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide (MTT) assay was used to screen for anticancer activity of the respective extracts, at varying concentrations, against MCF-7 (human breast adenocarcinoma) cells, HT-29 (human colonic adenocarcinoma) cells and HeLa (human cervical cancer) cells. All of the extracts showed cytotoxic activity in all three cell lines to varying extents, depending on the extract used and cell line screened. The acetone extract of A. afra proved to be the most effective inhibitor with the lowest IC50 (2.65 ± 1.05 μg/ml) having been shown in MCF-7 cells. A. afra and A. absinthium showed similar inhibitory patterns, with the methanol- and acetone extracts having been the most potent inhibitors of each of the respective cell lines in general. Fluorescence microscopy employing 4',6-diamidino-2-phenylindole dihydrochloride (DAPI) and propidium iodide (PI) staining indicated that the acetone extract of A. afra induces apoptosis in MCF-7 cells as apposed to necrosis, and the results were comparable to those obtained for cells exposed to cisplatin. Screening of the A. afra acetone extract for toxicity in normal human cells using the CellTiter-Blue® assay indicated the extract to be toxic to peripheral blood mononuclear cells (PBMC’s) at concentrations comparable to that for MCF-7 cells, while cell cycle analysis of MCF-7 cells exposed to the A. afra acetone extract indicated the extract’s ability to induce apoptosis comparable to that of cisplatin, with the extract exerting its activity at a point during or just prior to the S phase of the cell cycle.

The promotion and development of indigenous knowledge pertaining to the traditional African healing system is one of the prime objectives set out by the South African government. Despite excellent research opportunities and funding, the biggest problem with ethnopharmacological research is a lack of interaction with indigenous communities, which tends to dilute the benefits this research has to offer these communities. The primary aim of this study was thus to promote the traditional African healing system through collaborative medicinal plant research with local traditional health practitioners. The research collaboration aimed to validate some biological activities of traditional remedies used by collaborating traditional health practitioners and ensured interactive sessions where scientific literature, research practices, findings and relevant legislation were discussed and debated. The joint development of a medicinal garden was a valuable tool in realising these goals. Aqueous and ethanol extracts of Bulbine frutescens, Ornithogalum longibracteatum, Ruta graveolens, Tarchonanthus camphoratus and Tulbaghia violacea were selected for antimicrobial, anticancer and anti-diabetic screening, because of their sustainable utilisation potential. The ethanol extract of T. violacea produced the best antimicrobial activity on Bacillus subtilis (100% growth inhibition) and Candida albicans (89% growth inhibition) at 250 μg/ml. The EC50 for the ethanol extract of T. violacea against HT29 colon cancer cells was 101 μg/ml. The aqueous extracts of B. frutescens and T. camphoratus (0.5 and 50 μg/ml) produced the highest overall glucose uptake activity in Chang liver and C2C12 muscle cells. T. camphoratus was unanimously chosen by participating practitioners as the plant to be investigated further. The aqueous extract of T. camphoratus increased glucose uptake in C2C12 muscle cells through increased translocation of GLUT4 to the plasma membrane and activation of the PI3-kinase and AMP-kinase pathways. It produced some alpha-glucosidase inhibitory activity at concentrations of ≥ 200 μg/ml. Apart from interactive feedback seminars at which the findings were presented to participating practitioners, all scientific literature regarding the plants was summarised, translated, compiled and given to participating practitioners in written format. An indigenous knowledge agreement has been negotiated and will formalise the collaboration in future. It is recommended that future research focuses on plants with economic development potential that can be cultivated in the medicinal garden.

An ethnobotanical survey of plants used for the treatment of eye infections was carried out in the Eastern Cape Province of South Africa. The study revealed that 12 plant species are commonly used with Hippobromus pauciflorus (L.f) Radlk being the most frequent. The crude extracts of the leaves, stem bark and roots of the plant were investigated for antimicrobial activity against 10 bacterial and four fungal strains. The methanol extracts of the plant parts were the most active and showed appreciable activity against Gram-positive and Gram-negative bacteria. The methanol extracts of the leaves and stem bark inhibited the growth of fungi with activities ranging from 78.70 percent to 100 percent on Aspergillus niger and Penicillium notatum. The acetone extracts of the leaves and stem bark were active against A. niger (51.76 percent) and P. notatum (77.22 percent). The aqueous extract of Hippobromus pauciflorus leaves at 50, 100 and 200 mg/kg body weight doses were evaluated for anti-inflammatory, analgesic and antipyretic activities in male Wistar rats. Anti-inflammatory activity was studied by using carrageenan-and histamine induced oedema right hind paw volume while the analgesic effect was evaluated using formalin-induced pain and tail flick nociception response. The brewer’s yeast-induced pyrexia model was used for the antipyretic investigation. The extract at all the doses used significantly inhibited both the carrageenan- and histamine-induced inflammation in a manner that was not dose dependent. The extract reduced the formalin-induced pain licking as well as prolonged the reaction time in the tail flick-induced pain. The effects of the aqueous extract of the plant leaves at 50, 100 and 200 mg/kg body weight doses were investigated for 14 days on some biochemical parameters of male Wistar rats. The extract at all the doses tested did not significantly alter the levels of white blood cells, red blood cells, mean corpuscular volume, platelets, neutrophils, monocytes, lymphocytes and large unstained cells. While the levels of haemoglobin, packed cell volume and basophils increased at specific doses, those of mean corpuscular haemoglobin, mean corpuscular haemoglobin concentration and eosinophils decreased. In vitro antiviral activities of the plant against herpes simplex virus type 1(HSV-1) and coxsakie virus B6 were investigated. Cytotoxicity was evaluated by MTT assay in Vero cells. At concentrations ranging from 165-270/μg/ml there was significant antiviral activity against HSV-1, but toxicity was also noted. There was no antiviral activity against coxsakie virus B6. Bioactivity-guided fractionation of the leaves of H. pauciflorus yielded three known compounds. From the ethyl acetate fraction, two compounds were isolated (epicatechin and -sitosterol ) and from the n- hexane fractions, one compound (lupeol) was isolated. They were isolated and identified using various techniques. The antimicrobial, anti-inflammatory, analgesic and anti-pyretic activity of these compounds have been reported in literature. The structure and distribution of foliar appendages on the leaves of H. pauciflorus were examined by scanning electron microscope. The leaves have long unicellular nonglandular trichomes which were distributed over the mid rib and densely populated at the edges of the adaxial and abaxial surfaces. In general, the experiments and tests conducted in this study appear to have justified the use of Hippobromus pauciflorus for the treatment of eye infections and make a substantial contribution to the knowledge base of the use of herbal medicine for the treatment of the eye infections.

General Abstract There has been a steady increase in the use of medicinal plants in the last two decades in both developed and developing countries for prevention, management and treatment of diseases. This increase has been due to reasons such as ease of access, better cultural acceptability and compatibility, cost effectiveness and also the bid to “go natural”. Unfortunately, the notion that herbal medicines are relatively safe because they are natural has led to serious and potentially fatal consequences in phytotherapy. The lack of rigorous research to prove the effectiveness and safety of many medicinal plants is of great concern to the health care system. This thesis therefore addresses not just the efficacy, but also the safety of the extracts of the berry of Solanum aculeastrum - a medicinal plant used, among other things, for the treatment of breast cancer in the Eastern Cape Province of South Africa. Particular attention was paid to the possible effect of different processing methods of the berry extracts on inflammation, cytotoxicity, and toxicity. In studying the comparative effects of various processing methods, four different preparations of the extracts were investigated. These include fresh, dried, boiled fresh, and boiled dried berries. While the effect of processing on the anti-inflammatory properties of the extracts was not dose dependent, the percentage reduction in inflammation was highly significant and more prominent in both concentrations of the boiled fresh berries than the reference drug, indomethacin. Furthermore, the four extracts varied in their ability to act either centrally or peripherally in their effect on pain. Assessment of the analgesic response using the formalin test showed that, at both concentrations tested, none of the extracts inhibited the first phase of the formalin test. Furthermore, it was observed that boiling had differing results on the activity of the iii fresh and dried extracts. While boiling of the dried berries reduced pain in the rats, the opposite trend was observed with the boiled fresh berries. Results of the influence of processing of the berries on cytotoxicity indicated that the extracts are potent inhibitors of human breast, cervical and colonic carcinoma cells and the non-cancerous cells (both the actively dividing and confluent Chang liver cells). Although, in terms of relative potency, the fresh berries appeared to be the most active of the extracts, processing of the berries caused an increase in apoptotic cells and a subsequent decrease in the necrotic cells. The effect of processing on the safety of the berry of S. aculeastrum on the rats fed for 28 days was also investigated. The various doses (1, 10 and 25 mg/kg body weight) of all the four extracts did not alter the activity and the weight of the animals throughout the period of treatment. A reduction in organ to body weight ratio of the heart, kidney, liver and spleen was observed in all the extracts. Regarding the haematological parameters, different patterns of effect were observed between the extracts and within the treated doses. The observed alterations in the biochemical parameters by the various extract of Solanum aculeastrum berries at all the doses may have consequential effects on the normal functioning of these organs. In conclusion, this study has shown that there is some justification for the traditional use of the berries of Solanum aculeastrum in the treatment of inflammation related ailments and cancer. However, the medicinal use of the plant also poses considerable health risks. Investigation conducted into the plant’s safety revealed that the berry extracts were nephrotoxic, hepatotoxic, haematotoxic and at higher doses, fatal. Another concern with regard to the plant’s safety is the non-selectivity of its extracts in the inhibition of carcinoma, actively dividing and un-dividing cells. Assessment of the effect of the processing on the berry’s efficacy and safety as herbal iv remedy produced mixed results. On the one hand, processing seemed to improve the extract’s anti-inflammatory and analgesic activity, while reducing its cytotoxic potential. On the other hand, a reduction in the toxicity was observed on the processed extracts compared to the fresh ones. This may be an indication that processing has an overall beneficial effect on the medicinal properties of the plant and should thus be considered as a method of making the berries of Solanum aculeastrum safer for use as a herbal remedy.

Thesis (M.Sc. (Microbiology)) -- University of Limpopo, 2015 / Antioxidants play an important role in living organisms to control level of free radicals and other reactive molecules in the body to reduce oxidative damage. Synthetic antioxidant compounds are used in food industries as food additives to boost our immune systems. These compounds are associated with a number of critical side effects including liver damage and carcinogenesis. Scientists are also concerned about microorganisms that have developed resistant genes against current antibiotics used in hospitals. The aim of the study was to isolate and characterize bioactive compounds from Ricinus communis leaves with activity against Staphylococcus aureus (ATCC 29213), Enterococcus faecalis (ATCC 29212), Escherichia coli (ATCC 25922) and Pseudomonas aeruginosa (ATCC 27853). Consequently, medicinal plants are studied and considered for their efficacy and safety, because they possess bioactive compounds with various biological activities. Leaves of R. communis were collected at the University of Limpopo, Turfloop campus in Limpopo province, South Africa. The leaves were dried and milled to a fine powder. A number of trial extraction methods were employed using various solvents of different polarities on a fine powder leaves to identify the best extraction method. Plant extracts were analyzed by thin layer chromatography (TLC) developed in four mobile phases. To detect separated phytochemical compounds, TLC plates were sprayed with vanillin- sulphuric acid in methanol and heated at 110oC for optimal colour development. Qualitative antioxidant activity was determined by using 2, 2–diphenyl-1-picrylhydrazyl (DPPH) assay on TLC plates. Quantitative antioxidant activity was determined by measuring percentages scavenging activity of DPPH and 2, 2’-azinobis-(3-ethylbenzothiazoline-6-sulphonic acid (ABTS) free radical molecules by plant extracts. Antibacterial activity of all extracts was quantified by a serial microbroth dilution method while bioautography was used in qualitative analysis of the active compounds. Cytotoxicity effect of R. communis extracts was evaluated using tetrazolium-based calorimetric assay on human Caucasian skin fibroblast (Bud-8) cell line. Anti-inflammatory activity was assessed using phagoburst kit on Raw 264.7 macrophages cell line. Pure compounds were subjected to nuclear magnetic resonance spectroscopy for 1H, 13C and DEPT experiments to elucidate structures of compounds. 2 During extraction process, methanol was the best extractant, extracting greater amount of extracts than any of the other solvents. Serial exhaustive extraction method was selected as the best extraction method for extracting compounds from ground plant materials. In quantitative antioxidant assays, chloroform and methanol extracts had highest percentage scavenging activity against DPPH free radicals compared to other extracts and vitamin C. Methanol extract had the highest percentage scavenging activity of ABTS free radicals and minimum percentage scavenging activity was in hexane extract. Acetone, ethyl acetate and ethanol extracts showed strong antioxidant activity against DPPH free radicals in qualitative antioxidant assay on TLC plates. In quantitative antibacterial assay, crude extracts showed lowest minimum inhibitory concentration value of 0.13 mg/ml against all tested organisms and the highest was 1.05 mg/ml. Hexane extracts revealed potent antibacterial activity against all tested microorganisms on bioautograms. Hexane and acetone extracts also revealed anti-inflammatory activity and have ability to reduce oxidative stress. In cytotoxicity effect of plant extracts, Methanol extracts had lethal concentration for 50% of the cells (Lc50) of 784 μg/ml on Human Caucasian skin fibroblast (Bud-8) cell line while hexane extracts had Lc50 of 629 μg/ml. Plant extracts with high Lc50 are low toxic to normal cell line and preferable to work with for drug development. Bioassay-guided fractionations results in successful isolation of three antioxidant and two antibacterial compounds from R. communis using column chromatography. Isolated compounds were tested for their biological activities using qualitative DPPH assay on TLC plates for antioxidant activity and bioautography for antibacterial activity. Antioxidant compounds showed strong antioxidant activity after spraying with DPPH in methanol and antibacterial compounds showed less activity compared to the crude extracts. The study suggests the use of crude extracts to fight against pathogenic microorganisms compared to pure compounds. Compound 4 was successful identified as the mixture of stigmasterol and β-sitosterol. The present study recommends the use of R. communis leaves as the potential source of antioxidant, antibacterial and anti-inflammatory compounds. The study serves as a scientific proof for use of this plant in traditional medicine for treatment of various ailments.

Thesis (M. A. (African languages)) -- University of Limpopo, 2015

Please read the abstract in the section 00front of this document. / Dissertation (MSc (Pharmacology))--University of Pretoria, 2003. / Pharmacology / unrestricted

Thesis (Ph.D. Agriculture (Plant Production)) -- University of Limpopo, 2022 / The unique phytochemical composition of the medicinal plant cancer bush (Sutherlandia frutescens) have made its foliage to gain much attention in South Africa due to its health benefits. In situ harvesting of the plant parts of this important species serve as one potential strategy to avert its extinction through whole plant harvesting, a common practice by rural communities. However, such a strategy is limited by lack of information on the agronomic requirements of the plant species and its susceptibility to root-knot (Meloidogyne species) nematodes. The objectives of the study were four-fold, namely, to: (1) identify nodulation bacteria associated with wild S. frutescens using morphological and biochemical techniques, (2) assess the efficacy of the nodulation isolates from different centres of biodiversity of S. frutescens in Limpopo Province, South Africa (3) test the compatibility of cucurbitacin-containing phytonematicides on S. frutescens for managing population densities of Meloidogyne species and (4) determine the nutritional water productivity (NWP) of S. frutescens in association with water scarcity of the region where the plant species originated. In achieving Objective 1, nodules from S. frutescens roots were washed in distilled water and healthy, undamaged, firm and pink nodules were sterilised. Aseptic nodules from S. frutescens roots and commercial strains were transferred into a smasher biomerieux polythene bag containing 10 ml distilled water and crashed to produce a milky suspension the milky suspension was streaked on Yeast extract mannitol agar (YEMA). After gram reaction, colony characterisation includes the investigation of shape, colour, configuration, elevation and margin of bacterial colony as observed in colonies on nutrient agar plates of overnight grown microorganisms using a microscope. The medium for biochemical test was prepared, inoculated with 5 μl purified xxv bacterial cultures and incubated at 37°C for 48 h. Identification of the bacterial isolates was performed using VITEK 2 Systems (bioMérieux, Inc., North Carolina, USA). Using morphological and biochemical techniques, the bacterial species associated with roots of S. frutescens in the wild were assayed primarily those in the genera Raoutella ornithinolytica and Enterobacter cloacae species dissolvens. The VITEK 2 Systems confirmed the identification of the bacterial species from 80 to 96% of the samples. Three species were confirmed from another sampling area, Sphingomonas paucimobills, Raoutella ornithinolytica and Enterobacter cloacae species dissolvens from by 86 to 96% of the samples. In achieving Objective 2, the five treatments, namely, Bradyrhizobium spp. (Arachis) strain, Rhizobium leguminosarum strain, Tubatse strain, Sebayeng strain and untreated control, were laid-out in a randomised complete block design, with seven replications during the first season (Experiment 1) and with eight replications during the second season (Experiment 2). The seasonal interactions (Experiment 1 × Experiment 2) on plant and nutrient elements were not significant (P ≤ 0.05) and data for the two seasons were pooled (n = 75). Relative to untreated control, commercial (Bradyrhizobium and Rhizobium strain) and native strains (Tubatse and Sebayeng strain) significantly increased plant height by 31, 33, 44 and 40%, respectively, root length by 30, 41, 40 and 42%, respectively and dry shoot mass by 48,195 and 17%, respectively. Similarly, rhizobia strains significantly contributed to the increase in nitrogen assimilation by 7, 25 and 80%, respectively, protein synthesis by 13, 10, 24, 69%, respectively, and symbiotic efficiency by 31, 133, 292 and 82%, respectively. However, rhizobia inoculants had no significant effects on potassium and phosphorus in leaf tissues. In achieving Objective 3, in Mean Concentration Stimulation Point (MCSP) experiments, seven treatments, xxvi namely, 0, 2, 4, 8, 16, 32 and 64% for each phytonematicide, were arranged in a randomised complete block design (RCBD), with 8 replicates. In application interval experiments, treatments, based on “weeks-per-month-of-30 days” for M. javanica, which translated to 1, 2, 3 and 4 weeks, were arranged in a RCBD, with 10 replicates. Nemarioc-AL and Nemafric-BL phytonematicides had MCSP values of 3.43 and 4.03%, respectively, with the plant having high tolerance level to the products. The respective application interval of the two products for managing population densities of Meloidogyne species were 29 and 17 days. The dosage models for Nemarioc-AL and Nemafric-BL phytonematicides were 6.62 and 13.26%, respectively. In achieving Objective 5, the study used nine treatments designated as T1, T2, T3, T4, T5, T6, T7, T8 and T9, respectively, consisting of 1, 2, 3, 4, 5, 6, 7, 8 and 9 seedlings/hole of drip irrigation transplanted using a 3S planter under field conditions, arranged in randomised complete block design (RCBD) with 9 replications (n = 81) in two seasons. The NWP of total flavonoids, total tannin and total phenol exhibited positive quadratic relations in varied planting density suggesting that this cultural practices could be manipulated to improve NWP of cancer bush. In conclusion, the wild bacterial isolates, sampled from S. frutescens plant grown in the field, outperformed the commercial bacterial strains in enhancing the productivity of the test plants. The empirically established dosage model for Nemarioc-AL and Nemafric-BL phytonematicides could be used to control Meloidogyne species in cancer bush production. There is a need to further investigate the responses of the identified strains to the test phytonematicides. Findings of the study openend new frontiers in the development and commercialisation of the observed native bacterial strains for the cultivation of S. frutescens, which has excellent medicinal importance as a cure or management for cancer. / Agricultural Research Council-Universities Collaboration Centre, the National Research Foundation (NRF) and the Flemish Inter-University Council of Belgium

Plants are able to take up and accumulate certain environmental contaminants such as heavy metals. When the plants are ingested by man, these contaminants are transferred along the food chain. Due to the poorly regulated medicinal plant trade in South Africa, many opportunities exist for heavy metal contamination of medicinal plants namely contaminated harvest sites as well as poor drying, processing, storage, transport and manufacturing conditions. The concentrations of five heavy metals (As, Cd, Co, Ni, Pb) and six microelements (B, Cu, Fe, Mn, Mo, Zn) were determined in some commonly used South African medicinal plants obtained from street markets. Elemental content was determined using inductively coupled plasma optical emission spectrophotometry (ICP-OES). Some of the medicinal plant samples investigated contained As and Cd at levels exceeding the World Health Organization limits of 1 and 0.3 mg kg-1 respectively. Lead and Ni were detected in all the samples. Elevated Fe and Mn levels were recorded in certain plant species. The results revealed multiple metal contamination in some medicinal plant parts sold in local markets and is thus grounds for concern. The effects of Cd application on growth parameters of some medicinal plant species belonging to the Hyacinthaceae (Albuca setosa, Eucomis autumnalis, Eucomis humilis, Merwilla plumbea) gave insight into heavy metal accumulation and distribution in these species. Application of Cd at 5 mg l-1 over a 12 week period reduced growth in A. setosa. The medicinally used A. setosa bulbs accumulated 37 mg kg-1 Cd after 12 weeks. Cadmium application at 2 mg l-1 over a six week period had no effect on growth parameters of E. autumnalis or E. humilis. However, a substantial difference in total Cd accumulation was detected in the plants (40.2 and 15.3 mg kg-1 respectively). Cadmium application at 2 mg l-1 significantly reduced the fresh weight of leaves, bulbs and roots of M. plumbea. Although most of the Cd was stored in the roots, the medicinally used bulbs accumulated up to 11.6 mg kg-1 when applied at 10 mg l-1. The antagonistic effect between Cd and Zn treatments and their effect on micronutrient distribution in M. plumbea were investigated. Five treatments were evaluated: (1) Hoagland’s nutrient solution (HS) (control) (2) HS + Cd 2 mg l-1 (single) (3) HS + Cd 2 mg l-1 + Zn 50 mg l-1 (combination) (4) HS + Cd 2 mg l-1 + Zn 100 mg l-1 (combination) (5) HS + Cd 2 mg l-1 + Zn 150 mg l-1 (combination). Cadmium readily accumulated in leaves, bulbs and roots of M. plumbea when supplied at 2 mg l-1. Zinc at 50 mg l-1 led to increased Cd accumulation. However, further increases in Zn concentration showed an antagonistic effect of Zn on Cd uptake and accumulation. Thus, increasing Zn levels in soils may be favourable for reducing toxic Cd accumulation in M. plumbea plants. Boron was not significantly affected by the addition of Cd to the media. However, with an increase in Zn, leaf B content increased while the B content in the bulbs and roots decreased. Copper and Mo levels were not significantly affected by treatments with Cd or Cd/Zn combinations. Compared to the control, Cd and Cd/Zn applications caused an increase in Mn content in leaves, bulbs and roots. Iron levels of M. plumbea were not significantly affected by Cd in the media. However, with an increase of Zn in the Cd-containing media, Fe content in the leaves, bulbs and roots increased. Tulbaghia violacea is one of the few medicinal plants that is also frequently used as a leafy vegetable. Application of Cd at 2 and 5 mg l-1 to T. violacea of varying sizes (small 8 - 10 g, medium 16 - 20 g, large 80 – 95 g) elicited a difference in growth response, Cd accumulation and micronutrient distribution. Leaf length and fresh weight of leaves of the medium-size plants decreased with application of Cd at 2 mg l-1 whilst 5 mg l-1 Cd significantly decreased the number of leaves in small-sized plants. Small plants accumulated more Cd in the leaves than medium- or large-sized plants. Application of Cd at 2 mg l-1 and 5 mg l-1 lowered the leaf Cu, Fe, Mo and Zn contents in small- and medium-size plants. This study indicated that T. violacea has the ability to accumulate Cd. In addition, plant size plays an important role with regards to Cd accumulation and elemental distribution. The effect of various nutrient applications (10%, 50% and 100% Hoagland’s nutrient solutions (HS); and HS deficient in N, P or K) on growth parameters and micronutrient distribution in Dioscorea dregeana were investigated. Irrigating plants with 50% HS resulted in better growth performance, whereas a deficiency of either N, P or K negatively affected seedling growth. Plants grown in 10% HS contained higher total B, Fe and Mo levels compared to seedlings grown in 50% and 100% HS. Compared to the control, P deficiency resulted in a Fe increase in the leaves, tuber and roots while a lack of P and K significantly increased total Mn content in D. dregeana. The effect of excess Zn (100, 200 and 300 mg l-1) on growth performance, chlorophyll content and microelemental distribution on Dioscorea sylvatica was investigated. Growth parameters showed a significant decrease when supplied with Zn at 100 mg l-1. Zinc phytotoxicity was evident by the reduction in chlorophyll content. Highest Zn concentrations were detected in the roots. Certain micronutrients appear to be redistributed due to Zn toxicity. The effect of microelements (Cu, Zn) and heavy metals (Cd, Pb, Hg) on germination and seedling development of Bowiea volubilis, Eucomis autumnalis and Merwilla plumbea was investigated. Copper and Zn applied at 1 mg l.1 significantly reduced the percentage germination of E. autumnalis. Low concentrations (. 1 mg l.1) of Cu and Zn negatively affected the root growth of all three species. Mercury concentrations of 0.5 and 1 mg l.1 significantly decreased the percentage germination of B. volubilis and E. autumnalis respectively. Cadmium and Hg at 2 mg l.1 showed a negative effect on the root growth of B. volubilis. Concentrations of 0.5 mg l.1 of all heavy metals tested significantly decreased shoot length of M. plumbea. The effect of Cd on biological activity (anti-inflammatory, antibacterial and antifungal) of medicinal plants with previously confirmed activity was evaluated. When supplied with Cd at 2 mg l-1, Eucomis humilis bulbous extracts showed lower anti-inflammatory activity than the control for both COX-1 and COX-2 activity. Eucomis autumnalis bulbous extracts had greater COX-1 activity compared to the control. However, Cd suppressed the activity of COX-2. Compared with non-Cd-treated Merwilla plumbea plants (control), those supplied with Cd at 10 mg l-1 showed increased antibacterial activity against Bacillus subtilis, Klebsiella pneumoniae and Staphylococcus aureus. However, no change in activity against Escherichia coli was observed. Cadmium accumulation in the bulbs had no effect on antifungal activity of Tulbaghia violacea. Thus, optimized agricultural practices are essential for quality control of cultivated medicinal plants. The studies presented in this thesis collectively answer several questions related to heavy metal involvement in South African medicinal plants. The findings substantiate the need to regulate and monitor the South African medicinal plant trade against heavy metal contamination which will in turn provide a product of safety and quality to the consumer. / Thesis (Ph.D.)-University of KwaZulu-Natal, Pietermaritzburg, 2008.