Mmushi T MSc (Microbiology).pdf

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LIST OF TABLES Table 3.1.. Average MIC (mg/ml) and total activity (ml/g) of selected plant species after 24 hr incubation at 37 o C.................................................................. 91 Page Table 3.2. Average MIC (mg/ml) and total activity (ml/g) of selected plant species after 24 hr incubation at 37 o C.................................................................. 92 xvi
ABSTRACT The leaves of fifteen plant species were collected from the Lowveld Botanical Garden in Nelspruit, Mpumalanga Province, South Africa. The collection was based on a list of plants and their ethnopharmacological information provided by the Phytomedicine Programme at the University of Pretoria. The dried leaves of the plants were powdered and extracted using hexane, dichloromethane, acetone and methanol. The extracts were screened for antibacterial activity against Mycobacterium smegmatis and Rhodococcus erythropolis. The acetone extract of Milletia stulhimannii was the most active, showing activity against Mycobacterium smegmatis and Rhodococcus erythropolis with MIC values 0.13 and 0.08 mg/ml, respectively. Acetone extracts for all plants had the lowest MIC values ranging between 0.11-1.25 mg/ml and 0.08-1.25 mg/ml for M. smegmatis and R. erythropolis, respectively. Milletia stulhimannii, Albizia gummifera, Xanthocercis zambesiaca and Barringtonia racemosa extracts have shown the greatest potential for anti-tubercolosis agents. These were all active against M. smegmatis with an average MIC value of acetone extracts of 0.13 mg/ml. Apodytes dimidiata was selected for the isolation of active compounds since its activity on qualitative antibacterial activity assays was highly prominent on TLC plates in comparison to the other plant extracts. Two compounds were isolated from A. dimidiata but after purification, their MICs were above 2.5 mg/ml indicating a possible loss of activity during purification. The preliminary NMR spectra analysis suggested that the compounds were a long fatty acid and a triterpene. Future work is required to elucidate the chemical structures of the latter compounds and to test the activity of these compounds against Mycobacterium tuberculosis. xvii
Page 1 and 2: SCREENING, ISOLATION AND PURIFICATI
Page 3 and 4: DEDICATION I dedicate this work to
Page 5 and 6: TABLE OF CONTENTS Page Declaration
Page 7 and 8: 1.7 Microbiology and pathogenicity
Page 9 and 10: 6.2.1 Preparation of 1000 ml Middle
Page 11 and 12: LIST OF FIGURES Page Fig. 1.1. The
Page 13 and 14: Fig. 3.10. Fig. 3.11. Fig. 3.12. Fi
Page 15: Fig. 3.27. Fig. 3.28. Fig. 3.29. Fi
Page 19 and 20: CHAPTER ONE 1.1 Introduction Plants
Page 21 and 22: and R. erythropolis ATCC 4277strain
Page 23 and 24: Current therapeutic applications of
Page 25 and 26: agricultural (pesticides and herbic
Page 27 and 28: Fig. 1.3. The chemical structure of
Page 29 and 30: national pharmacopoeias, published
Page 31 and 32: Snyder and Kirkland (1979) tested t
Page 33 and 34: African medicinal plants. In certai
Page 35 and 36: susceptibility is determined by the
Page 37 and 38: equal volume of sterile distilled w
Page 39 and 40: Fig. 1.6. Albizia gummifera (Lemmen
Page 41 and 42: Fig.1.9. Apodytes dimidiata (The Wo
Page 43 and 44: 1.6.7. Kirkia acuminata Kirkia acum
Page 45 and 46: Fig. 1.14. (A) Maytenus udanta and
Page 47 and 48: from East Coast Fever, and people t
Page 49 and 50: 1.7. Microbiology and pathogenicity
Page 51 and 52: indicates that essential oils can a
Page 53 and 54: Rhodococcus erythropolis is an aero
Page 55 and 56: 1.9. Aim and objectives 1.9.1. Aim
Page 57 and 58: each of the finely ground samples a
Page 59 and 60: phytochemical analysis (section 2.4
Page 61 and 62: The sub-fractions of 90:10 and 85:1
Page 63 and 64: 3.2. Phytochemical analysis of plan
Page 65 and 66: BEA EMW CEF DCM HEX ACE MET DCM HEX
Page 67 and 68:
BEA EMW CEF DCM HEX ACE MET DCM HEX
Page 69 and 70:
BEA EMW CEF Fig. 3.8. Chromatograms
Page 71 and 72:
BEA EMW CEF DCM HEX ACE MET DCM HEX
Page 73 and 74:
3.4. Minimum inhibitory concentrati
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Table 3.2.Average MIC (mg/ml) and t
Page 77 and 78:
BEA EMW CEF HEX DCM ACE MET HEX DCM
Page 79 and 80:
Page 81 and 82:
Page 83 and 84:
BEA EMW CEF Fig. 3.19. Bioautograms
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BEA EMW CEF . Fig. 3.21. Bioautogra
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Of the two solvent systems used fro
Page 89 and 90:
F1 F2 F3 F4 F5 F6 F7 F8 F9 F10 F11
Page 91 and 92:
F1 F2 F3 F4 F5 F1 F2 F3 F4 F5 F1 F2
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Fig. 3.28. TLC profile showing the
Page 95 and 96:
Fig. 3.30. TLC profile of fractions
Page 97 and 98:
Fig. 3.32. TLC profile of pooled su
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Fig. 3.34. 1 H NMR spectra of compo
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Page 103 and 104:
Page 105 and 106:
Fig. 3.40. 13 C NMR spectra of comp
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Page 109 and 110:
Page 111 and 112:
CHAPTER FOUR DISCUSSION Tuberculosi
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e that active compounds may be in v
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this plant. However, as can be seen
Page 117 and 118:
Britton, G. 1991, Methods in Plant
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Eloff, J.N. 1999a. Which extractant
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Harborne, J.B., and Williams, C.A.
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Kemper, K.J. 1999. Longwood Herbal
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Middleton, E., Chithan, K. 1993. Th
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Pretorius, S.J., Joubert, P.H., Sco
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Van der Geize R., and Dijkhuizen, L
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CHAPTER SIX APPENDICES 6.1. Appendi
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121 o C and 1 atm. For preparation
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