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.

Thesis (M.Sc. (Biochemistry )) --University of Limpopo, 2007 / Refer to document / The Cannon Collins Trust Fund and the National Research Foundation

Many South African medicinal plants face the threat of over-collection for use in traditional medicines. Many bulbous plants suffer as the whole plant is removed from the wild so that the bulb may be used for medicine. Micropropagation is a technique which can be used as an alternative to conventional propagation methods. Micropropagation produces many plantlets in a relatively short period of time. Different plant parts of Brunsvigia undulata F.M. Leight, a rare South African species of medicinal value, were used in an attempt to produce in vitro plantlets using micropropagation techniques. Although leaf and floral explants were successfully formed from seedling explants and twin-scales. Seeds germinated quickly in culture. Seedlings which grew from seeds were cut into sections and used to initiate bulblets. Seedling explants formed bulblets, shoots and callus best when the explants included a meristematic region. Callus from seedling explants formed shoot clusters readily when placed on hormone-free MURASHIGE and SKOOG (1962) (MS) medium. Shoots from shoot clusters formed bulblets and rooted on medium supplemented with IBA. The greatest rooting response was achieved by bulblets on 1 mgl-1 IBA. The callus which was left after shoot clusters were separated was placed back onto hormone-free MS medium. Callus explants continued to form shoot clusters. Twin-scales, cut from large parent bulbs, were cultured on 25 hormone treatments. Bulblets formed on twin-scales even in the absence of plant growth hormones. Bulblets formed by twin-scales were used to determine the effects of both medium constituents and environmental factors on bulblet multiplication. Bulblet multiplication was greatest when bulblets were split in half and cultured as half-bulblets. Optimal multiplication was achieved on hormone-free MS, with 4% sucrose, kept at high temperatures in the dark. Bulblets were successfully initiated and multiplied from both seedlings and twin-scales. Bulblets which were produced via both protocols were acclimatized relatively easily. Both explant types could be used to mass propagate Brunsvigia undulata. / Thesis (M.Sc.)-University of KwaZulu-Natal, Pietermaritzburg, 2009.

South Africa is home to a wide diversity of cultural groups, all of which utilize the flora for a variety of purposes. This is true with regard to traditional medicine systems which are similar to those of the rest of Africa south of the Sahara, with diviners (sangomas) and herbalists (inyangas) as the key health providers. In addition, the Country is rich in plant diversity with some 30 000 species of flowering plants - almost one tenth of the worlds recorded higher plants. This incorporates a large diversity of plants including trees, shrubs, herbs, bulbs and corms. The adverse effects of traditional medicinal plants and natural products are not well documented in the literature. Recently, many plants used as food or in traditional medicine have been shown to be potentially mutagenic using in vitro assays. Thus, the scientific evaluation of traditional medicine and medicinal plants is very important to validate claims made on safety and efficiency of such usages. After a survey of the available ethnobotanical literature, ten trees used in South African traditional medicine were selected. These species were: Acacia niolotica subspecies kraussiana, Acacia sieberiana, Albizia adianthifolia, Combretum kraussii, Faidherbia albida, Ficus sur, Prunus africana, Salix mucronata, Terminalia sericea and Trichilia dregeana. Plant parts including leaf, root and bark were collected from each of the selected trees (exceptions were Albizia adianthifolia, Faidherbia albida, Terminalia sericea and Prunus africana) and extracted using ethyl acetate, ethanol and water individually to ensure the extraction of compounds over a wide range of polarities. The extracts (in total, 78) were screened for antibacterial, anti-inflammatory (COX-1 and COX-2) and antiacetylcholinesterase activities and investigated for their potential mutagenic effects using the Ames test. Antibacterial activity was detected using the disc-diffusion and microdilution assays. The extracts were tested against Gram-positive bacteria: Bacillus subtilis, Staphylococcus aureus, Micrococcus luteus and Gram-negative bacteria: Escherichia coli and Klebsiella pneumoniae. Of the 78 different plant extracts 111 tested (final amount of plant material was 1 mg per disc), 84% showed activity against Gram-positive bacteria. From this percentage, 20% also showed activity against Gram-negative bacteria. The best inhibition was observed with ethyl acetate and ethanol root extracts of Terminalia sericea against both Gram-positive and Gram-negative bacteria. In the micro-dilution assay, 55% of the plant extracts showed minimum inhibitory concentration (MIC) values ~ 1.56 mg/ml against Gram-positive and/or Gram-negative bacteria. The ethyl acetate bark extract of Acacia sieberiana and the root and bark ethyl acetate extracts of Acacia nilotica inhibited bacterial growth of both Gram-positive and Gram-negative bacteria at concentrations ~ 0.8 mg/ml. The aqueous leaf extracts of Acacia sieberiana had a low MIC value (0.3 mg/ml) against Gram-negative Kleibsiella pneumoniae and the ethyl acetate extracts of the root inhibited growth of Escherichia coli with an MIC value of 0.1 mg/ml. However, these two extracts showed no activity in the disc-diffusion assay. The MIC values of the neomycin (control) were 0.8 I-Ig/ml and 3.1 I-Ig/ml against Kleibsiella pneumoniae and Escherichia coli respectively. In the anti-inflammatory test, 70% of the plant extracts from different plant parts (leaf, root, bark) of the tree investigated showed strong inhibition in both the CQX-1 and CQX-2 bioassays. The CQX-2 inhibitory effects of aqueous extracts were generally lower when compared to the organic solvent extracts. However, water extracts of Acacia nilotica was an exception (~ 90%). In the acetylcholinesterase inhibitory test, 21% of the plant extracts were active at concentrations ~ 1 mg/ml using the micro-plate assay. The lowest IC50 value was 0.04 mg/ml obtained with an ethanol bark extract of Combretum kraussii. The IC50 value of the galanthamine (positive control) was 2 I-IM. None of the investigated plants showed any potential mutagenic effects with Salmonella typhymurium strain TA 98 using the Ames test. Using bioassay-guided fractionation, anolignan B was isolated from the ethyl acetate root extract of Terminalia sericea. Antibacterial activity of anolignan B was determined using the microdilution assay. The compound possessed activity against both Gram-positive and Gram-negative bacteria. The lowest MIC value (3.8 IJg/ml) was observed with Staphylococcus aureus. MIC value of the neomycin was 1.5 IJg/ml. Anti-inflammatory activity of anolignan B was detected using the CQX-1 and CQX-2 bioasays. The compound showed strong inhibitory activity against CQX-1 and weaker activity against CQX-2. The ICso values were 1.5 mM and 7.5 mM with CQX-1 and CQX-2 respectively. The ICso values of indomethacin were 0.003 mM and 0.186 mM against CQX-1 and CQX-2 respectively. There were no potential mutagenic effects showen by anolignan B against Salmonella typhimurium strain TA 98 in the Ames test. Isolation of anolignan B from Terminalia species and the antibacterial and anti-inflammatory activities observed in this work have not been reported previously and could therefore be recorded as novel biological activities for this compound. These results also support the idea that the use of ethnobotanical data can provide a valuable short cut by indicating plants with specific uses which might likely be sources of biologically active chemicals. / Thesis (Ph.D.)-University of KwaZulu-Natal, Pietermaritzburg, 2005.

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.

Throughout the history of mankind, many infectious diseases have been treated with plant extracts. Venereal infections are one such group and are regarded as conditions that are highly responsive to traditional treatment. Aqueous, ethanol and ethyl acetate extracts of 13 plants used in South Africa for the treatment of venereal diseases were screened for in vitro antibacterial, antifungal, mutagenic and antimutagenic activities. Antibacterial activity was evaluated using the disc-diffusion and microdilution assays to determine the minimal inhibitory concentration (MIC) values of the extracts. The extracts were tested against the Gram-positive bacteria Bacillus subtilis and Staphylococcus aureus, and the Gram-negative bacteria Escherichia coli and Klebsiella pneumoniae. Among the plants tested, Gunnera perpensa, Harpephyllum caffrum, Hypoxis latifolia and Ledebouria ovatifolia showed the best antibacterial activity. The aqueous rhizome extract of Gunnera perpensa displayed good activity against Gram-negative bacteria with an MIC value of 0.78 mg/ml, and against S. aureus (0.78 mg/ml). Aqueous and ethanol extracts of H. caffrum bark were active against both Gram-positive and Gram-negative bacteria. Hypoxis latifolia aqueous corm extracts exhibited very good MIC values against K. pneumoniae (0.78 mg/ml), E. coli and S. aureus (1.56 mg/ml). Ethanol and ethyl acetate bulb extracts of Ledebouria ovatifolia displayed good activity against Bacillus subtilis bacteria with MIC values of 0.78 mg/ml and 0.39 mg/ml respectively. Antifungal activity was evaluated using the microdilution bioassay. Good activity was shown by the ethanolic bark extracts of Bersama lucens and Harpephyllum caffrum against Candida albicans. Only in the case of Harpephyllum caffrum did aqueous extracts have activity against Candida albicans. In the Ames test, all plant extracts showed a negative genotoxic response except for ethanol and ethyl acetate bulb extracts of Cyrtanthus obliquus which induced mutations in TA98. Moderate antimutagenic activity was observed with the ethyl acetate extract of G. perpensa and the ethanolic extract of H. latifolia. High antibacterial and antifungal activity detected with Harpephyllum caffrum bark extracts resulted in an investigation on seasonal and geographical variation of this inhibitory activity. Seasonal variation in antibacterial and antifungal activities was investigated in order to determine the best collection time to ensure potential high medicinal activity in plant preparations. The highest inhibitory activity was detected with plant material collected in June and December 2003, with a decline in activity when collections were made in September 2004. The chemical profiles of TLC chromatograms were compared and little variation was found, particularly in the case of plant material obtained from the Botanic Garden of the University of KwaZulu-Natal and a 'Muthi' Shop in Pietermaritzburg. Identification of active compounds from G. perpensa and H. caffrum was not successful due to insufficient amounts of isolated fractions. / Thesis (Ph.D.)-University of KwaZulu-Natal, 2006.

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.

The rhizomes of Alepidea amatymbica and Alepidea natalensis are used for medicinal purposes. Because of the increase in demand for these plants the species is becoming scarce. As the seed biology of neither species is well defined, conditions as well as treatments required for optimum germination and vigour were studied. Seeds were exposed to various physical factors such as varying light and temperature conditions and cold stratification, sowing depth and seed storage. The effects of smoke-water, butenolide (3-methyl-2H-furo [2, 3-c] pyran-2-one) a novel smoke compound and chemical substances (gibberellins, kinetin and KNO3) were also tested in order to improve seed germination. Alepidea amatymbica and A. natalensis achieved the highest seed germination (72.5% and 80%, respectively) at 25 °C under a 16 h photoperiod with a mean germination time (MGT) of 18 and 12 days, respectively. Phytochrome studies showed that A. natalensis requires light for germination. Cold stratification (5 °C) for 14-28 days significantly improved the percentage germination of both species (> 90%) compared to non-stratified seeds (control) at 25 °C under a 16 h photoperiod. Sowing A. amatymbica and A. natalensis seeds at a depth of 0.5 cm resulted in higher percentage germination compared to 2.5 cm. The highest emergence rate for A. amatymbica was 40% at a sowing depth of 0.5 cm and the lowest emergence rate was 3% at 2.5 cm. Six months storage of A. natalensis seeds at room temperature (25 ± 2 °C) showed maximum germination (99%) with a MGT of 9 days. Smoke-water treatment of A. amatymbica seeds significantly enhanced germination from 72% to 91%. Smoke and butenolide at 10 °C and 25 °C promoted germination of A. natalensis seeds in a 16 h photoperiod. Smokewater application significantly improved both germination and seedling vigour of A. natalensis. GA3 (10-8 M) was the best treatment for achieving maximum percentage germination of A. natalensis seeds. Antibacterial (two Gram-positive bacteria: Bacillus subtilis, Staphylococcus aureus and two Gram-negative bacteria: Escherichia coli, Klebsiella pneumoniae), antifungal (Candida albicans), anti-inflammatory (COX-1 and -2) and genotoxicity tests (Ames test) were carried out on petroleum ether (PE), dichloromethane (DCM), 80% ethanol (EtOH) and water extracts of the two Alepidea species. Water extracts of A. natalensis rhizomes exhibited high activity (MIC values of 0.78 mg/ml) against the four bacterial strains. High activity was also observed in the PE and DCM leaf extracts of the same plant against the Gram-positive bacteria. The PE and DCM extracts of A. amatymbica rhizomes exhibited the best activity (MIC values of 0.39 mg/ml) against Bacillus subtilis. The rest of the extracts showed low activity (MIC values >1 mg/ml). All the extracts showed activity against Candida albicans, with A. natalensis leaf extracts exhibiting the highest antifungal activity with MIC values of 0.88, 0.20 and 0.78 mg/ml for PE, DCM and EtOH, respectively. EtOH extracts had inhibition less than 40% for both A. natalensis and A. amatymbica. All the PE extracts showed higher inhibitory activity for COX-2 than for COX-1. PE and DCM extracts had percentage inhibitions above 70% in both COX-1 and COX-2 assays. The Ames test for genotoxicity revealed that none of the plant extracts were genotoxic to the Salmonella TA98 tester strain. / 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.

The post-harvest physiology of nine frequently used indigenous southern African medicinal plants was investigated, in particular the effects of storage time and accelerated ageing on the biological activity and chemical constituents of these plants. Water, ethanol and hexane extracts of fresh plant material as well as material that had been stored in dry form in paper bags at room temperature for 90 days (short-term) were tested. Three bioassays, the COX-1 anti-inflammatory assay, nematode anthelmintic assay and minimum inhibitory concentration anti-bacterial assay, were used to determine biological activity. Thin layer chromatography of all the plant extracts were used to determine changes in chemical composition. The plants tested were Alepidea amatymbica Eckl. & Zeyh., Leonotis leonurus (L.) R. Br., Drimia robusta Bak., Vernonia colorata (Willd.) Drake, Scilla natalensis Planch., Eucomis autumnalis (Mill.) Chitt. subsp. autumnalis, Bowiea volubilis Harv. ex Hook. f., Helichrysum cymosum (L.) D. Don and Siphonochilus aethiopicus (Schweinf.) B. L. Burtt. Only those plants, which are known to exhibit a particular biological activity either traditionally or scientifically, were tested in the relevant bioassays. Of the plant extracts tested for anthelmintic activity only the water extracts showed activity and very little change in activity was observed after storage. Of the plant extracts tested for anti-inflammatory activity the ethanol extracts generally yielded highest activity. S. natalensis and B. volubilis both showed an increase in cyclooxygenase inhibition (anti-inflammatory) activity after storage whereas S. aethiopicus, H. cymosum, D. robusta and V. colorata showed a loss in activity after storage. The anti-inflammatory activity of E. autumnalis did not change. The water extracts of plants tested for antibacterial activity showed no activity, whereas the ethanol extracts generally showed an increase in activity. The TLC fingerprints indicated that there was chemical break-down during storage in certain species. These corresponded to the changes in biological activity. Alepidea amatymbica, Eucomis autumnalis, Helichrysum cymosum, Leonotis leonurus, Siphonochilus aethiopicus and Vernonia colorata were investigated further as to the effect of one year's storage (long-term storage) on their chemical composition and biological activity. Similar trends to that of the 90-day storage were observed. Activity gained in plants that were stored for 90 days was retained after a year of storage. Elevated temperature and humidity (55 C and 100% relative humidity) were used to accelerate the ageing process of Alepidea amatymbica, Leonotis leonurus and Vernonia colorata plant material. Again changes in the chemical composition and biological activity were observed, and the extent of these changes was greater than those in the stored material. The compounds responsible for the cyclooxygenase inhibition in the ethanolic extracts of Alepidea amatymbica leaf material appear to be stable and were not affected by the conditions of the accelerated ageing procedure (55 C and 100% humidity for seven days), but the root material lost activity, as did the leaf material of Leonotis leonurus. The leaf material of Vernonia colorata showed a slight (8%) increase in cyclooxygenase inhibition activity. The response of the plant material to accelerated ageing with respect to antibacterial activity varied with plant species. Alepidea amatymbica root material and Vernonia colorata leaf material appear to be stable whereas the other plant materials lost activity after prolonged (25 days) ageing. / Thesis (M.Sc.)-University of Natal, Pietermaritzburg, 2003.