Lung infections in cystic fibrosis (CF) patients are very serious and are usually fatal. Prolonged antibiotic treatment can result in the development of antibiotic resistance. Thus, studies are required to determine if traditional medicinal plants have any significant effects (i.e. antibacterial effects in this case) on certain bacteria causing lung infections in CF patients, providing possible alternative treatment. The aim of this study was to test the effects of several Eastern Cape medicinal plants on specific bacteria causing lung infections in CF patients, for possible antibacterial activities. Four plants traditionally used to treat lung infections (Artemisia afra, Dodonaea angustifolia, Leonotis leonurus & Tulbaghia violacea), and different strains of four bacteria (Staphylococcus aureus, Pseudomonas aeruginosa, Pseudomonas species & Klebsiella pneumoniae), usually causing lung infections were investigated. Methanol, aqueous and acetone extraction methods were used to make extracts of the various plants chosen. A decoction of each plant was also investigated. An adapted microtitre plate method for testing the antibacterial activity of the plant extracts against the chosen bacteria was used. The minimum inhibitory concentration (MIC) of each plant extract was determined by means of this method.

Artemisia afra is one of the oldest, most well known and widely used traditional medicinal plants in South Africa. It is used to treat many different medical conditions, particularly respiratory and inflammatory ailments. There is no reported evidence of its use for the treatment of cancer but due to its reported cytotoxicity, an investigation of the mode of cell death induced by an ethanol A. afra extract using two cancer cell lines was done. IC50 values of 18.21 and 31.88 μg/mL of ethanol extracts were determined against U937 and HeLa cancer cells, respectively. An IC50 value of the aqueous extract was greater than 250 μg/mL. The ethanol extract was not cytotoxic against confluent control cell lines, Chang Liver and Vero cells. The effect of the cytotoxic ethanol A. afra extract on U937 and HeLa cells and their progression through the cell cycle, apoptosis and mitochondrial membrane potential was investigated. After 12 hours of treatment with A. afra a delay in G2/M phase of the cell cycle was evident. Apoptosis was confirmed using the TUNEL assay for DNA fragmentation, as well as fluorescent staining with annexin V-FITC. Apoptosis was evident with the positive control and A. afra treatment at 24 and 48 hours. JC-1 staining showed a decrease in mitochondrial membrane potential at 24 hours. It was deduced that A. afra ethanol extract induces caspase-dependent apoptosis in a mitochondrial dependent manner. Plants harbour many compounds that are not only useful to the plants but also to mankind. Many metabolites have been isolated from A. afra and their biological activity characterised. Due to observed apoptosis induction, isolation of cytotoxic compounds was done and a new sesquiterpene lactone from A. afra was isolated. Structural elucidation of the compound was done by IR, 1D and 2D NMR, CD and mass spectrometry and it was identified as isoalantolactone. HeLa cancer cells were treated with isoalantolactone and cytotoxicity was exhibited in a dose-dependent manner. A low IC50 value of 8.15 ± 1.16 μM was achieved. This study showed that isoalantolactone is partly responsible for the observed A. afra cytotoxicity. Due to the evidence of G2/M arrest, the anti-mitotic potential and the possible onset of mitotic catastrophe by A. afra and isoalantolactone was investigated. It was evident from various flow cytometric analysis of cyclin B1 and phospho-H3 and confocal microscopy that A. afra does possess anti-mitotic activity by causing hyperpolymerisation of tubulin and cells progress into the mitotic phase where M arrest is experienced. The anti-inflammatory activity of sesquiterpene lactones is well documented; however, the anti-inflammatory activity of A. afra is not. Here, it is reported that the production of NO and COX-2 protein levels in RAW 264.7 cells decrease in the presence of A. afra and isoalantolactone after stimulation with LPS. The activated NF-κB subunit, p65 was also investigated. The results suggest that A. afra and isoalantolactone inhibit p65 activation as a decrease in the activated subunit was evident. Thus, the results indicate that exposure to A. afra and isoalantolactone induces an anti-inflammatory response. In conclusion, this study shows, for the first time, the mechanism of induced apoptosis, the anti-mitotic and anti-inflammatory activity of A. afra and its isolated compound, isoalantolactone. It also proves that although extensive research may have been done on a particular plant, as with A. afra, more can be discovered leading to the identification of new compounds and integration of signalling pathways that can be exploited for the treatment of various diseases and ailments.

A mini dissertation submitted in partial fulfillments for the degree of master of Environmental Education in the department of Comparative and Science Education at the University of Zululand, 2006. / Most tribes in South Africa still make use oftraditional medicines and use prescriptions ofherbs and other natural materials (Matsiliza and Barker 2001). Medicinal plants from Northern KwaZulu-Natal (KZN), particularly areas such as UIundi, Empangeni, Eshowe Mtubatuba and Nongoma are no exception and are a constituent of a traditional culture. The informal trade in indigenous plants for medicinal use (umuthi) is growing constantly (Mander, 1998). There are many reasons for this, including an expanding population coupled with modernisation of society that is hastened by the development of roads, improved communication methods and the migration of people from villages to cities. The prohibitive cost of modern medicines, together with the high rate of unemployment is forcing many people to revert to traditional medicines and even to gather and sell medicinal plants as a living. This results in over-exploitation of some plant species, sometimes to the brink ofextinction (Matsiliza and Barker 2001). This study aims at investigating the implications of plant harvests that are sold at bus ranks in northern KwaZuluNatal, and at identifYing target species for threatened plant programmes with the help of data gathered via direct interviews with medicinal plant sellers. It also aims at identifying and remedying knowledge gaps that may exist among plant gatherers about propagation techniques applicable to the medicinal plants they harvest.

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

Mycobacterium tuberculosis and Human Immuno-Deficiency Virus (HIV) have a high prevalence in South Africa. The development and spread of drug resistant tuberculosis is a serious problem which is exacerbated by tuberculosis (TB) co-infection in HIV patients. Traditional medicinal plants like Annona muricata and Artemisia afra are used for respiratory ailments and antiviral therapies respectively. The aim of this study was to evaluate Annona muricata (ethanolic extract) and Artemisia afra (ethanolic and aqueous extracts) for inhibitory activities against M. tuberculosis and HIV. In vitro bioassays for anti-TB activity included: microplate alamar blue assay (MABA), flow cytometry and ρ-iodonitrotetrazolium chloride assays while anti-HIV activity was determined using an HIV-1 reverse transcriptase colorimetric ELISA kit and an HIV-1 integrase colorimetric immunoassay. Cytotoxicity of plant extracts were assessed by the MTT assay on Chang Liver and HepG2 cells. Potential synergistic effects were determined using the basis of Combination Index. Potential interactions of plant extracts with drug metabolic pathways were evaluated with the Glutathione-S-Transferase assay kit as well as the CYP3A4 assay kit. A. muricata ethanolic extract exhibited anti-TB activity with MIC 125 μg/mL. MABA was shown to be the most sensitive and effective method for the detection of anti-TB activity. Artemisia afra aqueous extract showed HIV-1 reverse transcriptase inhibition exhibiting ˃85 percent inhibition at 1 mg/mL while the ethanolic extracts of A. afra and A. muricata showed inhibition of HIV-1 integrase activity at ˃86.8 percent and ˃88.54 percent respectively at concentrations >0.5 - 4 mg/mL. The aqueous extract of A. afra displayed inhibition of HIV-1 integrase ˃52.16 percent at 0.5 mg/mL increasing to 72.89 percent at 4 mg/ml of the extract. A. muricata was cytotoxic at an IC50 of 30 μg/mL and 77 μg/mL on Chang Liver and HepG2 cells respectively, whilst A. afra aqueous and ethanol extracts were not cytotoxic to both cell lines. The ethanolic extract of A. muricata showed both antagonistic and synergistic properties at various IC values, when used in conjunction with rifampicin. A. afra ethanolic extract interrupted GST activity while aqueous extracts of A. afra and A. muricata had a slight effect. All extracts interrupted CYP3A4 activity, however the ethanolic extracts of A. muricata and A. afra showed greater inhibition than the aqueous extract of A. afra. These extracts should be investigated further as they could be an important source of compounds for treatment of M. tuberculosis and HIV respectively.

The phytochemistry of Ochna serrulata (Hochst.) Walp. was investigated for the first time; two new dimeric chalcones (5-deoxyurundeuvine C and serrulone A) and two new biflavonoid derivatives (4,4’,7-tri-O-methylisocampylospermone A and 4”’-de-Omethylafzelone A) were isolated. These compounds were isolated along with the known compounds lophirone A, afzelone B, campylospermone A, isocampylospermone A, ochnaflavone, 2”,3”-dihydroochnaflavone, lophirone C, psilosin, 3’-O-methylpsilosin, a cyanoglucoside, epicatechin, (2’,4’- dihydroxyphenyl)acetic acid, methyl (2’,4’-dihydroxyphenyl)acetate, irisolone 4’- methyl ether, iriskumaonin 3’-methyl ether, 3',4'-dimethoxy-6,7-methylenedioxyisoflavone, lophirone L, syringaresinol and 16α,17-dihydroxy-entkauran-19-oic acid. The growth inhibitory effect of these compounds was evaluated against three cancer cell line panel of TK 10 (renal), UACC62 (melanoma) and MCF7 (breast) using a sulforhodamine B (SRB) assay. Ochnaflavone and 3’-methoxypsilosin demonstrated selectivity and only inhibited the growth of melanoma cancer cells. However, ochnaflavone showed higher activity by totally inhibiting the growth of melanoma cancer cells at 12.91 μM, whereas, 3’-O-methylpsilosin has this effect at a concentration of 14.11 μM. Lophirone C, a dimeric chalcone, demonstrated the highest cytotoxic activity amongst all isolated compounds against renal, melanoma and breast cancer cells with TGI at 35.63 μM, 11.67 μM and 30.35 μM, respectively. Lophirone A, a rearranged biflavonoid, showed TGI against these cancer cells at 58.96 μM, 26.23 μM and 40.01 μM, respectively. The rest of the compounds showed no significant cytotoxicity against the three cancer cells. The new biflavonoid, 4,4’,7-tri-O-methylisocampylospermone A demonstrated the highest antimalarial activity against chloroquine-resistant strains of Plasmodium falciparum (FCR-3) with IC50 of 11.46 μM, followed by ochnaflavone (17.25 μM). iv Serrulone A (26.52 μM), lophirone A (29.78 μM), 5-deoxyurundeuvine C (31.07 μM), lophirone C (35.31 μM), 4”’-de-O-methylafzelone A (38.43 μM), afzelone B (39.54 μM), irisolone 4’-methyl ether (40.72 μM) and syringaresinol (42.66 μM) were moderately active. The following compounds exhibited the lowest antimalarial activity, 2”,3”-dihydroochnaflavone (61.86 μM), iriskumaonin 3’-O-methyl ether (93.69 μM),3’- O-methylpsilosin (106.35 μM) and16α,17-dihydroxy-ent-kauran-19-oic acid (106.48 μM). Owing to the observed and reported biological/pharmacological activity, ochnaflavone (an ether-linked biflavone consisting of apigenin and luteolin moieties) was selected for synthetic studies. An older method, nucleophilic aromatic substitution (SNAr) was successfully applied in the construction of the diary ether. Oxidative ring cyclization of the ether-linked dimeric chalcone was achieved by using heated pyridine and iodine. The two methods can be extended further in the synthesis of other novel biflavones with ether linkage. / Thesis (Ph.D.)-University of KwaZulu-Natal, Pietermaritzburg, 2012.

The broad objective of the study was to determine and validate medicinal plants used by resource-limited farmers to control internal and external parasites in goats in the Eastern Cape Province, South Africa. A survey was conducted among 50 farmers and three herbalists to determine medicinal plants used to control parasites in goats. The survey revealed nine plant species belonging to eight families that were used. Among the identified plant species, Aloe ferox, Acokanthera oppositifolia and Elephantorrhiza elephantina were the plants having the highest Fidelity Level for their use, each scored 100.00 percent, followed by Albuca setosa (83.33 percent). These plants were then selected for validation studies. Gas-Chromatography-Mass-Spectrometry (GC-MS) revealed 7, 33, 26 and 32 bioactive phytochemicals in A. ferox, E. elephantina, A. oppositifolia and A. setosa, respectively. Terpenes and fatty acids were present, oxygenated terpenes being the most abundant hydrocarbons present in all the four plant species. The effect of acetone, methanol and ethanol extracts of leaves of Aloe ferox and Acokanthera oppositifolia on tick repellency and acaricidal activity were investigated on blood engorged Ambylomma hebraeum and Rhipicephalus decoloratus ticks at concentration 15, 30 and 50 percent. The 30 and 50 percent acetone extract of A. ferox and Dazzel dip had the highest acaricidal properties of 100 percent. The 50 percent methanol extract of A. oppositifolia and 50 percent acetone extract of A. ferox had the highest repellency activity of 89 percent and 85.33 percent, respectively. Results from this study revealed that the efficacy of medicinal plants used by farmers to control ticks vary with the type of solvent used for extracting the bioactive compounds. Furthermore, it revealed that Aloe ferox and A. opppositifolia plant extracts possess repellent and acaricidal activities. In a study to investigate the anthelminthic effect of crude extracts of Elephantorrhiza elephantina and Albuca setosa plants, significant anthelminthic effect on nematodes was observed in both plants. In this study, all E. elephantina and A. setosa extracts caused paralysis and mortality. Methanol was the most effective solvent in extracting bioactive compounds and methanol extract showed the best anthelminthic effects among the crude extracts investigated in both plants. The least time taken for the worms to be paralysed was 8.33 mins and 14.33mins in 100mg/ml methanol extracts of E. elephantina and A. setosa, resepectively. Methanol extract of E. elephantina and A. setosa (100mg/ml) had the highest anthelminthic activity and mortality was recorded after 18mins and 20mins, respectively. Results from this study revealed that these two plants possess anthelminthic activities. The study revealed that resource-limited farmers use medicinal plants to control internal and external parasites in goats. Gas-Chromatography-Mass-Spectrometry analysis showed that these plants contain bioactive compounds that have a potential in controlling parasites. Validation studies showed that A. ferox and A. oppositifolia possess repellent and acaricidal activities whereas A. setosa and E. elephantina possess anthelminthic activities.

Ph.D. (Chemistry) / The Forensic Chemistry Laboratory of Johannesburg (FCL JHB) is tasked with the chemical analysis of a variety of samples to assist in determining the cause of death where unnatural cause is suspected. Some of the samples submitted to the laboratory have a herbal or muti connotation, but a large portion of these cases turn out to have no herbal components present as only pharmaceutical or agricultural products are detected in these samples. This study combined, for the first time, forensic investigation, chemistry and botany to create a unique platform needed for the identification of poisonous plants and their components in forensic exhibits and viscera. The research was focussed on the poisonous plants previously detected at the laboratory, as well as the requests received for the analysis of muti/toxic plant components. The selection of plants included Nicotiana glauca, Datura stramonium / Datura ferox, Callilepis laureola, Boophone disticha / Ammocharis coranica, Abrus precatorius, Ricinus communis, Nerium oleander / Thevetia peruviana and Bowiea volubilis. All these species are known to have caused fatalities, hence their choice. Nicotiana glauca has been implicated in the deaths of at least 15 people since 2001. It was previously detected by GC-MS (EI) in plant exhibits, but could not be detected in a viscera matrix. A selective extraction method for alkaloids was used to extract botanical and viscera samples. Anabasine was successfully detected on the HPLC-MS (EI) system but this detection technique was not considered sensitive enough. A very sensitive HPLC-MS method was developed on the ZMD detector by using electrospray technology. This method outperformed both electron impact detectors (GC and HPLC) and could detect 1ng/ml anabasine with relative ease in full scan mode. Datura stramonium and D. ferox have not been previously positively linked to any human poisoning or death due to exposure to botanically derived products at the FCL JHB. Atropine and scopolamine were successfully ionised in ESI positive mode and could be detected at 10 pg/ml and 100 pg/ml level respectively. The identities of the compounds were confirmed by characteristic ISCID fragmentation patterns. The developed method was successfully applied to a suspected heart attack case. The results proved conclusively that the deceased was given D. ferox seeds as part of his meal and an overdose of atropine and scopolamine contributed to his death...

Type II diabetes is becoming a growing problem in developed countries worldwide. The median age for diagnosis was around sixty, but recent surveys have shown that the entire age distribution curve shifting left. The incidence of type II diabetes is thought to be parallel with the growing rate of obesity associated with an unhealthy western diet. Type II diabetes is an expensive disease to manage, it is for this reason that cheaper medication needs to be investigated in the form of traditional plants, such as Sutherlandia frutescens. Prescription medication, such as tricyclic antidepressants, may also increase body weight thereby playing a role in obesity. The cause of weight gain in such cases may go unrecognized or lead to cessation of the medication with or without the practitioner’s knowledge or approval. It is therefore necessary to investigate the causative agents responsible for the excessive weight gain. Drinking water containing extracts of S. frutescens or metformin was administered to two groups of eleven insulin resistant male Wistar rats. The insulin resistant control group received water without any medication. Rats were sacrificed after 8 weeks allowing for fasting blood glucose, insulin and tissue glycogen content determination. Glucose uptake was also determined using [3H] deoxyglucose. The effect of the medication and the diet on muscle post receptor insulin signaling proteins was determined through Western blots. Liver proteomics was also performed using 2-D electrophoresis. In a separate experiment 26 male Wistar rats were exposed to strepotozotocin toxin, 7 of these rats received intravenous insulin treatment, 7 rats received S. frutescens extract and 7 rats received a combination of both medications, the remaining 5 received no treatment and served as the control. Rats were sacrificed after 6 days allowing for fasting blood glucose, insulin and tissue glycogen content determination. Two groups of 14 male Wistar rats received amitriptyline or trimipramine (common tricyclic antidepressants) in their drinking water, the control group (30 rats) received water without any medication. The rats’ weight and food consumption was monitored throughout the trial and their oxygen consumption was also determined. Rats were sacrificed after 6 weeks or 14 weeks of medicinal compliance allowing for fasting blood glucose, insulin and tissue glycogen content determination. Glucose uptake was also determined using [3H] deoxyglucose. S. frutescens treatment normalized circulating serum insulin levels and significantly increased the rate of glucose clearance. Certain post receptor insulin signaling proteins were also significantly increased relative to the insulin resistant control group. 2-D electrophoresis identified the normalization of protein levels associated with the urea cycle. S. frutescens was also able to, independently; maintain normoglycaemic levels in the strepotozotocin treated group. The tricyclic antidepressants significantly increased blood glucose levels while significantly reducing tissue glycogen levels for both sacrifice periods. Serum insulin remained unchanged while a significant increase in insulin degradation and insulin degrading enzyme levels were found for both antidepressants. S. frutescens shows promise as a low cost antidiabetic medication for future use. Although the antidepressants did not promote weight gain, the increase in blood glucose levels may be cause for concern in patients with a pre-disposition toward developing diabetes.

It is estimated by the World Health Organization (WHO) that by 2040 neurodegenerative disorders will collectively surpass cancer as the primary cause of death in industrialised countries (WHO,2006). Natural flora represents one of the most important therapeutic sources in modern drug discovery, however only a limited number of plant species have been screened for their neuroprotective value. The neuroprotective potential of eleven Algerian and two South African medicinal plant extracts were assessed in this study, aiming to identify promising candidates for future research. Neurodegenerative disorders such as Parkinson’s disease are characterised by distinct biochemical features, including protein misfolding/-aggregation, excessive oxidative stress and apoptotic cell death. As such, medicinal plant extracts were screened for biological properties directly relevant to neurodegeneration. The capacity to induce autophagy was also investigated as mounting evidence suggests that activation of this pathway may reduce abnormal protein aggregation and promote neuronal survival.