MSc (Microbiology) / Department of Microbiology / BACKGROUND: Medicinal plants form an important part of the Southern African cultural heritage. Indigenous populations, for example the Vha-Venda people, tend to use medicinal plants in formulations rather than western medicines for health and survival. In order to certify and give scientific credibility to the use of medicinal plants formulations used by Vha-Venda people for the treatment of diseases, several assays were carried out. The present study was aimed at assessing phytochemical content, antimicrobial, antioxidant and cytotoxic activities of four indigenous Venda medicinal plants in a formulation and compare their activity with each plant used individually. METHODS: Peltophorum africanum (roots), Pterocarpus angolensis (bark), Terminalia sericea (roots) and Ximenia caffra (roots) were collected from the Thohoyandou area. The collected plant parts were extracted with methanol and water respectively. Individual plant extracts and Five designed formulations were tested for their antimicrobial activity against Staphylococcus aureus ATCC 25923 (Methicillin Resistant), Staphylococcus aureus ATCC 33591(Methicillin Susceptible), beta lactamase producing Klebsiella pneumonia (ATCC 700603) and extended spectrum beta lactamase producing E. coli (ATCC 35218), four clinical isolates of Candida spp and Cryptococcus neoformans using the Broth dilution method. Minimum bactericidal concentration (MBC) of the extracts was determined by culturing the contents of minimum inhibitory concentration (MIC) on nutrient agar. Similarly, minimum fungicidal concentration (MFC) was also determined by culturing contents of MIC in sabouraud dextrose agar (SDA). Extracts were further assessed for their total phenolic content, total flavonoid content and Qualitative phytochemical analysis. The antioxidant ability of the plants extracts and formulations to scavenge free radical DPPH was also determined. The plant formulations were assessed for their anti-HIV activity using the reverse transcriptase colorimetric assay kit. Cytotoxicity against human lymphatic endothelial cells (HLEC) was determined using MTT assay. RESULTS: Methanolic and aqueous extracts of T. sericea exhibited the best antifungal and antibacterial activities whilst P. angolensis and X. caffra showed poor activities. Methanolic plant formulations showed good activities compared to aqueous formulations. However, Fractional Inhibition Concentration Index showed that there was 1 synergistic interaction, 25 additive interactions and 14 antagonistic interactions between the plant extracts. The methanolic formulation 3 showed the best overall phenolic content at 11.85±0.109 mgGAE/g whilst aqueous X. caffra extract showed the least content at 4.546±0.104 mgGAE/g. Higher total flavonoid contents were seen in methanolic formulation 4 at 2.75±0.02 mgQE/g. Qualitative phytochemical analysis revealed the presence of flavonoids, phenolics, terpenoids, tannins, saponins and steroids in 80% of the tested plant extracts and formulations. All plant extracts and formulations exhibited good antioxidant activity against DPPH. The methanolic formulation showed the best antioxidant activity with IC50 of 0.094 ± 0.33μg/ml. For anti- HIV inhibition, all formulations at 200μg/ml exhibited higher percentage of HIV-1 reverse transcriptase inhibition with methanolic mixture 3 being the best overall at 97.5% activity whilst aqueous mixture5 was the least active with 63.03% inhibition activity. Moreover, the best anti-HIV activity at 100μg/ml was exhibited by methanolic mixture 3 at 71% inhibition. Furthermore, aqueous X. caffra, mixture 2 inhibited 26% and 51% at 12.5mg/ml and 3.125mg/ml respectively. Peltophorum africanum and mixture 5 inhibited 34%, 54% and 43% at 3.125mg/ml, 6.25mg/ml and 12.5 mg/ml respectively of Human Lymphatic Endothelial cells growth. CONCLUSIONS: The results from the study indicated that most of the commonly used traditional medicinal Plants in the Venda region when mixed together have merit for use in traditional medical practice as they have shown good antimicrobial activities, good antioxidant xviii activities, good phytochemical activities and good cell proliferation activity. However some formulations showed antagonistic interaction against bacteria. Some Individual medicinal plants showed toxicity at higher concentrations against immune cells. Whereas formulations promoted cell proliferation, therefore, the use of such individual plants in the treatment of infections should be highly monitored as they may pose a health threat to normal immune cells. Generally, plants are potential pharmacological agents which needs to be preserved and harvested with care. / NRF

MSCAGR (Plant Production) / Department of Plant Production / Indigenous trees play important roles in livelihoods for rural communities. Sclerocarya birrea and Berchemia discolor are indigenous in Africa and are used in rural communities for food and livestock feed, as well as for medicinal and construction purposes. These trees are subject to attack by insects, which can result in lower tree productivity. However, there is no documented information about insects found on both tree species in South Africa. This study investigated the seasonal abundance and diversity of insects on S. birrea and B. discolor in Tshikundamalema Area, Limpopo Province, South Africa. Five trees were selected for each of the two tree species at two sampling sites with different vegetation cover, one site mainly the woodland bushveld while the other site was savannah grasslands. Canopy fogging was used to sample insects. Insect samples were collected at one time point for each of the seasons: summer (February), autumn (May), winter (July) and spring (November). Insects were identified to morphospecies and Simpson’s diversity index was used to compare insect diversity on both trees. Analysis of variance was used to compare the abundance of insects across seasons. The most abundant insects were collected from Coleoptera, followed by Hemiptera, Hymenoptera, Diptera, Lepidoptera and Orthoptera. A total of 3259 insects belonging to the six Orders were collected. All specimens belonging to Coleoptera, Hemiptera and Hymenoptera (3216 in total) were identified to Family level, with a total of 97 morphospecies, belonging to 19 Families. Insects from Diptera, Lepidoptera and Orthoptera were not identified further due to their low numbers. The abundance and diversity of insects were affected by seasons, sites and tree species. The number of insects was high during the spring season with 1782 insects, and summer coming second with 1104 individuals, followed by autumn (238 insects), then winter (92). B. discolor sampled for a great number of insects (1741) as compared to S. birrea (1475). The woodland bushveld (1924 insects) surpassed the savannah grassland (1292 insects) site in terms of the insect numbers sampled. / NRF

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MSCAGR ( Plant Production) / Many high valued tree species of medicinal significance in the Limpopo Province, South Africa exhibit seed dormancy, and also contain aromatic oils which inhibit rooting of their stem cuttings. These plant species are under pressure due to human over-exploitation. The main objective of this study was to investigate effects of rooting hormones on the rooting ability that will help in domesticating some of the selected high valued medicinal plants, Elaeodendron transvaalense (bushveld saffron), Brackenridgea zanguebarica (yellow peeling plane), and Warburgia salutaris (pepper-bark tree). Stem cuttings of these plant species were prepared and treated with various concentrations (500, 1000 and 2000 ppm) of IBA, IAA and NAA in different growth media (Natural soil, farm soil and hygromix) at a nursery house. 180 experimental units were sown and arranged in Randomized Complete Block Design (RCBD), each treatment replicated five times and then monitored for a period of three months. Data were only obtained from Brackenridgea zanguebarica as other species dried up two weeks after sprouting. The two variables measured from the experiments were sprouted stems and number of leaves. The data obtained were subjected to analysis of variance and least significant difference (LSD) at 5% probability level was used to compare treatment using STATISTICA software analysis package. The hormone, hormone concentration, growth media and their interactions had effect on sprouted stems and number of leaves produced on Brackenridgea zanguebarica cuttings, with no record of rooting ability. IBA (500 ppm and 1000 ppm) and control (without rooting hormone) showed high significant results with natural soil and farm soil in terms of leaf production compared to the hygromix, which is significantly lower from others. IBA at the various concentrations (500, 1000 and 2000 ppm) and the control gave the highest percentage sprouted stem on both natural soil and farm soil as compared with other hormones at the same / NRF

MSc (Botany) / Department of Botany / Since time immemorial plants have traditionally been used for various purposes in many parts of the world including Vhembe district, Limpopo province of South Africa. The use of C. abbreviata plants at Matsa village of Vhembe district also contributes significantly to the livelihood of the community. However, little work has been made in the past to properly document and promote the knowledge. The purpose of the present study was to record and analyse the ethnobotanical knowledge of C. abbreviata within the people of Matsa village around Nzhelele in Vhembe district. Semi-structured questionnaires were administered on 50 informants from different households selected randomly to gather data regarding the local name of C. abbreviata, the uses and its availability. Significantly higher number of medicinal use was reported by elderly people including both men and women as compared to middle-aged, younger people. Other use categories included firewood, furniture and joinery production and shade for people in the area. Awareness on the importance of the species should therefore be created amongst the people, especially the young ones. Understanding the ethnobotanical knowledge of indigenous species within an area is crucial towards development of its management plan. / NRF

PhD (Botany) / Department of Botany / Brackenridgea zanguebarica Oliv. (Ochnaceae) commonly known as “Yellow Peeling Plane”, is an important medicinal plant species, and one of the most threatened tree species that is endemic to Thengwe village in Vhembe District, South Africa. It is widely harvested for traditional purposes and difficult to propagate sexually and asexually. Nowadays, it has become threatened and this unique plant population is believed to be declining in the Brackenridgea Nature Reserve (BNR). According to the Red List of South African plants, B. zanguebarica is considered a critically endangered medicinal plant. All these facts combined with a lack of knowledge on the conservation of this species prompted this study on its conservation. To ascertain the effective conservation and proper management of this multipurpose tree, this study was conducted with the aim to improve its conservation strategy by investigating current threats to the existence of B. zangueberica, the chemical constituents from the different plant parts, the propagation methods of the species, and the factors delaying the germination of its seeds. In this study, an assessment of the current population threats was important as a first step in conservation; this would update the status of the species in its natural habitat. The current status of Brackenridgea zanguebarica was conducted through an extensive field survey in the Brackenridgea Nature Reserve (BNR), where a belt transect method was deployed to record quantitative information, while the harvesting damage was estimated using a sliding scale from 0 to 5. The structure of the populations in terms of the stem diameter size classes showed a bell shape vi pattern. About 60% of individuals showed some signs of plant parts extraction as the stem bark was reported to be the main plant part harvested. The population structure, however, was dominated by juvenile plants due to the selective harvesting of mature individuals; this hinders fruit production leading to poor or slow seed production. The present study aimed to compare the chemical constituents and the antimicrobial activities of some parts of this plant species. The extracts were screened for phytochemicals using standard methods. Thin-layer chromatography (TLC) was carried out to compare the chemical constituents using various solvent systems of varying polarity. Agar well diffusion and broth microdilution methods were used to evaluate the antimicrobial activity and the minimum inhibitory concentrations (MIC) of plant extracts respectively, against three bacteria and four fungi. The phytochemical analysis revealed a close similarity of various phytoconstituents of the three plant parts (stem bark, twigs and leaves). TLC analysis showed a slight difference in the acetone extract of different plant parts with more bands on the leaves than the stem bark and twigs. Acetone extracts of the three plant parts exhibited varying degrees of antimicrobial activity against Staphylococcus aureus and Candida glabrata, while none of the extracts showed any activity against Escherichia coli, Klebsiella pneumonia, Candida albicans, C. krusei, and C. parapsilosis. However, analysis of the antibacterial activity of various extracts revealed that the best inhibitory activity was produced by the stem bark extract compared to the leaf and twig extract. vii The sustainable way to meet the growing demand of Brackenridgea zanguebarica is to increase its availability through propagation practices. Unfortunately, the conservation of B. zanguebarica is hampered by lack of information on how it can be propagated. Sexual and asexual propagations were conducted to determine an efficient method to increase the number of individuals of this plant species. Seed propagation was carried out to study the effect of various pre-treatments on germination, in order to understand the germination requirements; vegetative propagation was conducted to assess if B. zanguebarica could be successfully propagated via stem cuttings if the appropriate growth hormones and growing media were applied. Brackenridgea zanguebarica seed did not germinate at all under any of the conditions tested. However, the results, showed the potential of propagating this species from stem cutting despite the poor results obtained. Thus B. zanguebarica was found to be difficult to propagate sexually and asexually. Seeds serve as a means of reproduction and a vital element to ensure the survival of plant species. Seed germination is controlled by a number of factors. The propagation of B. zanguebarica via seed is very difficult and research has not been undertaken to understand possible factors that may delay its germination. This study was aimed to investigate and describe both the morphological and anatomical features of B. zanguebarica seed, in order to identify structural features implicated in its poor germination. To achieve this goal, a morphological and anatomical study was conducted based on the observation of seeds under a scanning electron microscope (SEM) and light microscopy (LM). Water uptake potential was assessed by the increase in seed mass. The morpho-anatomical analysis indicated that some structures of the seed, such as seed coat, and the presence of endosperm viii surrounding the embryo that consisted of two prominent cotyledons, seem to greatly hinder the germination of this species. The research findings provided information regarding the seed structure which is one important step in identifying useful techniques to improve seed germination for conservation purposes. / NRF