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UNIVERSITY ------------------ OF -------------------- JOHANNESBURG THE ETHNOBOTANICAL, ANTIMICROBIAL AND PHYTOCHEMICAL SCREENING OF SELECTED MEDICINAL PLANTS FROM GA- MASHASHANE, LIMPOPO PROVINCE, SOUTH AFRICA BY LESIBA ABRAM PAPO Dissertation submitted in fulfilment of the requirements for the degree o f Masters in Botany In Botany and Plant Biotechnology Faculty of Science University of Johannesburg Supervisor: Professor A Moteetee Co-supervisor: Professor S van Vuuren July, 20 17 DECLARATION I, Lesiba Abram Papo, student number: 200935782, solemnly declare that this dissertation is my own original work. It is being submitted for the degree of Master of Botany at the University of Johannesburg, South Africa. This work has not been submitted before for any degree or examination and where sources are used in the study, they have been well referenced. Lesiba Abram Papo Date i DEDICATIONS I would like to dedicate this study to my grandfather, Elias Moshemane Papo and my beloved grandmother, Raesetsa Rachel Papo. To my grandfather, thank you for being the mouthpiece and the source of this indigenous information documented in the study. Your gentle spirit, guidance and the care you bestowed upon the entire family have always been the chief cornerstone to draw our strength to endure. To my grandmother, I have no words, you clothed us with your love, your gentle spirit and compassion and taught us to fix our eyes to the God above, and for that, thank you for shaping me and raising me into the person that I am today. Ke tloga ke leboga ditlou!! ii ACKNOWLEDGEMENTS ❖ Blessed be the name of God, our Heavenly father for providing me with the strength, opportunity, and time to start and seal the study. ❖ I would like to thank my supervisor, Professor Annah Moteetee. You have always being a mother to me and the rest of the Taxeth group. Your ability to see the good and the potential in us, and also to nurture us with your wisdom. It is an honour to work under your supervision, for you to grant us opportunities to challenge ourselves and for you to modify us into successful individuals, thank you for everything and above all for your Ubuntu. ❖ To my co-supervisor, Professor Sandy Van Vuuren, thank you for the tireless assistance and guidance with the antimicrobial screening. ❖ To my informants, thank you for your warm-hearted souls and providing me with your invaluable ethnobotanical information, your co-operation is highly appreciated. ❖ Malakia Manamela, Inter Letjeku and Johannes Mantjiu, thank you for assisting me with some plants collection and surveys, your efforts are highly appreciated. ❖ Mutsinda Ramavhunga, thank you for assistance with the study map area. ❖ To the Papo and Makuya family, thank you for your prayers, for the love, care and believing in me that I can make it. ❖ To my beloved mother, Josephine Papo, thank you for everything and moulding me into a success. Your love and words of encourangement are highly appreciated. in ❖ To my colleagues, Lerato Seleteng-Kose, Phumzile Madondo and the rest of the Taxeth group, thank you for being part of the academic family and strengthening each other through tough times. ❖ My beloved sister, Mpho Papo, National Research Foundation and the University of Johannesburg, thank you for financial assistance to make this study possible. ❖ Moloko Dikgale, Katlego Riet, Lesetja Kgobe, and Karabo Robaya thank you for being the brothers who kept me focused throughout. ❖ To my beloved lady and friend, Abigail Lethuba, thank you for the love, care, compassion and your ability to constantly keep me happy and focused. Your presence in my life is indeed a blessing. IV CONFERENCE PRESENTATIONS ❖ Papo, L. A., van Vuuren, S.F., and Moteetee A.N., 2016. The ethnobotanical, antimicrobial and phytochemical screening of selected medicinal plants from Ga- Mashashane, Limpopo Province, South Africa. Podium presentation at the joint conference of the 42nd annual SAAB and 12th bi-annual SASSB conferences held at University of the Free State, 10-13th January, 2016. ❖ Papo, L. A., van Vuuren, S.F., and Moteetee A.N., 2016. The ethnobotanical, antimicrobial and phytochemical screening of selected medicinal plants from Ga- Mashashane, Limpopo Province, South Africa. Oral presentation for the 5th postgraduate centre, Research and innovation cross-faculty symposium held at University of Johannesburg, 13th October 2015. ❖ Papo, L. A., van Vuuren, S.F., and Moteetee A.N., 2016. The ethnobotanical, antimicrobial and phytochemical screening of selected medicinal plants from Ga- Mashashane, Limpopo Province, South Africa. Oral presentation at the postgraduate Botany Symposium held at University of Johannesburg Lapa, 27th October, 2015. v ABSTRACT Medicinal plants are widely recognised and acknowledged in rural areas and globally they contribute to the healthcare system within various countries. The core indigenous knowledge on the use of these medicinal plants resides in elderly people and it is traditionally passed from one generation to the next through oral transmission, and often not efficiently documented. The Ga-Mashashane area, situated in the Capricorn District of the Limpopo Province is no exception to the risk of losing some of this information through oral transmission. However, ethnobotanical studies have been conducted at District level within the Province covering Waterberg, Capricorn, Sekhukhune, Mopani and Vhembe. The aim of the current study was to explore and document the indigenous knowledge on the use of medicinal plants by the people in the Ga-Mashashane area and to determine the efficacy of these medicinal plants through antimicrobial and phytochemical screening. Knowledge on the use of medicinal plants was gathered through structured questionnaires, which considered socio-demographics, plant organs used, dosage, and administration method. Forty-five medicinal plants were recorded to treat various human ailments in the area, and the majority of these plants were noted from Anacardiaceae, Asteraceae, Fabaceae, and Verbenaceae. Twelve new uses were recorded in the current study with Senecio barbertonicus Klatt (used as a purgative), recorded for the first time. Priority was given to gastrointestinal ailments treated with 22 medicinal plants, followed by respiratory ailments (18), skin ailments (16), and sexually transmitted infections (eight), while non-infectious ailments are treated with four species. Peltophorum africanum was the most highly utilized medicinal plant to treat all infectious ailments, followed by Acokanthera oppositifolia, Carissa bispinosa, Carpobrotus edulis, and Eucalyptus VI camaldulensis used to treat three infections. Oral administration, inhalation, soaking, topical ointment application, and smoke therapy were recorded as the preferred method for administration of the medicinal plants. The plants were further dried and ground for pathogen-specific antimicrobial screening against 16 pathogens of the gastrointestinal, respiratory, skin, and sexually transmitted infections, as well as qualitative phytochemical screening. Plants with outstanding noteworthy actitities were recorded from the organic extracts of Ozoroa sphaerocarpa (MIC range 0.1 - 1.0 mg/ml) and Searsia lancea (MIC range 0.1 - 1.0 mg/ml) against all six gastrointestinal pathogens. The organic extract of Peltophorum africanum exhibited potent activity against two respiratory pathogens (Klebsiella pneumoniae and Staphylococcus aureus both at 1.0 mg/ml) while the organic extract of Ozoroa insignis demonstrated excellent activity against Gardnerella vaginalis and Neisseria gonorrhoea at 1.0 mg/ml and 0.1 mg/ml respectively. Interestingly, the organic extracts of eight medicinal plants highlighted to treat STIs exhibited noteworthy activity against N. gonorrhoea at MIC range of 0.1 - 0.8 mg/ml. Antimicrobial screening against skin pathogens resulted in the organic extracts of Asparagus racemosus (MIC range of 0.1 - 1.0 mg/ml), Peltophorum africanum (0.3 - 1.0 mg/ml), and Ximenia caffra (0.5 -1.0 mg/ml) exhibiting potent activity by inhibiting all four pathogens known to cause skin infections. Noteworthy activity with essentials oils were recorded from Eucalyptus camaldulensis (1.0 mg/ml) and Lippia javanica (1.0 mg/ml) when screened against S. aureus. Phytochemical screening was performed on plants with newly recorded uses and previously deficient phytochemical information. Zanthoxylum humile emerged as the only plant with all screened phytochemicals; alkaloids, coumarins, flavonoids, glycosides, tannins, terpenoids, and saponins. In addition the aqueous extract of Z. humile demonstrated the lowest MIC value of 0.75 mg/ml against K. pneumoniae. Englerophytum magaliesmontanum and Ozoroa sphaerocarpa