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Analysis of Antibacterial Activity of Terminalia Sericea and Combretum Imberbe Two Combretaceae Species from Namibia a Thesis S

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Analysis of Antibacterial Activity of Terminalia Sericea and Combretum Imberbe Two Combretaceae Species from Namibia a Thesis S ANALYSIS OF ANTIBACTERIAL ACTIVITY OF TERMINALIA SERICEA AND COMBRETUM IMBERBE TWO COMBRETACEAE SPECIES FROM NAMIBIA A THESIS SUBMITTED IN PARTIAL FULFILLMENT OF THE REQUIREMENT FOR THE DEGREE OF MASTER OF SCIENCE IN MICROBIOLOGY OF THE UNIVERSITY OF NAMIBIA BY MAPIYE SAMSON (200970551) APRIL 2019 MAIN SUPERVISOR: PROF. KAZHILA C. CHINSEMBU CO-SUPERVISOR: PROF. MARIUS HEDIMBI ABSTRACT The study was conducted to investigate the antibacterial activities of stem and leaves from Terminalia sericea and the live and dead bark from Combretum imberbe. The aqueous and organic crude extracts of the two plants were tested on Gram-negative bacteria Escherichia coli, Shigella sonnei, Serratia marcescens, Helicobacter pylori and Gram-positive bacteria Enterococcus faecalis, Staphylococcus aureus and Clostridium perfringens using the disk diffusion method. All the crude extracts exhibited activity against the tested bacteria species except for extracts from C. imberbe (dead bark). Organic extracts had high activity with minimum inhibitory concentration (MIC) values of 0.1mg/ml for all plant parts against S. aureus , S. marcescens, and H. pylori whilst the aqueous extracts from T. sericea (stem and leaves) and C. imberbe (live bark) had MIC values of 1mg/ml all against S. marcescens and H. pylori. Fractions from the two plants were obtained by vacuum liquid chromatography (VLC) using different solvents with increasing polarity. The organic extract fractions from both plants were observed to have good activity against two of the seven bacterial species that were tested. The dead bark plant extracts from C. imberbe had MIC value of 10mg/ml with 100% Ethyl acetate extracts against the bacteria H. pylori. T. sericea leaf extracts had MIC value of 0.01mg/ml against H. pylori in 100% MeOH fractions whilst the stem extracts had activity at 0.01mg/ml against E. faecalis in 100% Acetone fractions. The results indicate that the two plants have good antibacterial activity hence further research is needed to isolate and identify active compounds which can be developed into effective drugs. ii TABLE OF CONTENTS ABSTRACT ............................................................................................................... II LIST OF TABLES ................................................................................................. VII LIST OF FIGURES .............................................................................................. VIII LIST OF ABBREVIATIONS ................................................................................. XI ACKNOWLEDGEMENTS ................................................................................... XII DEDICATION ....................................................................................................... XIII DECLARATIONS ................................................................................................ XIV CHAPTER 1: INTRODUCTION ........................................................................... 15 1.1 Background ................................................................................................ 15 1.1.1 Antimicrobial resistance ........................................................................... 16 1.2 Statement of the problem ................................................................................. 17 1.3 Objectives ........................................................................................................ 17 1.4 Significance of the study .................................................................................. 18 1.5 Limitations of the study ................................................................................... 18 CHAPTER 2: LITERATURE REVIEW ............................................................... 19 2.1 Medicinal plants ............................................................................................... 19 2.2 Antimicrobial ethno medicinal plants used in Namibia ................................... 20 2.3 Medicinal use of Combretaceae species in Namibia ....................................... 22 2.4 Secondary metabolites ..................................................................................... 26 2.4.1 Alkaloids ................................................................................................... 26 2.4.2 Anthraquinones ......................................................................................... 27 iii 2.4.3 Flavonoids ................................................................................................. 28 2.4.4 Terpenoids ................................................................................................. 29 2.4.5 Lectins ....................................................................................................... 30 2.4.6 Tannins ...................................................................................................... 31 2.5 Antimicrobial use ............................................................................................. 32 2.5.1 Antimicrobial resistant bacteria ................................................................ 33 2.5.2 Global burden ............................................................................................ 34 2.5.3 Burden in Africa ....................................................................................... 34 2.5.4 Burden in Namibia .................................................................................... 35 2.6 Pathogenic bacteria .......................................................................................... 35 2.6.1 Helicobacter pylori ................................................................................... 35 2.6.2 Staphylococcus aureus .............................................................................. 39 2.6.3. Pathogenic Escherichia coli ..................................................................... 42 2.6.4 Enterococcus faecalis (E. faecalis) ........................................................... 44 CHAPTER 3: MATERIALS AND METHODS ................................................... 47 3.1 Plant material collection .................................................................................. 47 3.1.1 Preparation of crude extracts..................................................................... 48 3.1.2 Test Bacteria ............................................................................................. 49 3.1.3 Preparation of plant extracts solution........................................................ 49 3.1.4. Susceptibility test ..................................................................................... 50 3.1.5. Minimum inhibitory concentration (MIC) ............................................... 50 3.1.6. Preparation of fractions ............................................................................ 51 3.1.7. Data analysis ............................................................................................ 52 CHAPTER 4: RESULTS ........................................................................................ 53 4.1 Antimicrobial assay of crude extracts .............................................................. 53 iv 4.1.1 Antibacterial activity of aqueous extracts at 100mg/ml............................ 53 4.1.2 Antibacterial activity of organic extracts at 100mg/ml ............................. 54 4.2. Minimum inhibitory concentration (MIC) ...................................................... 55 4.2.1 Antibacterial activity of aqueous crude extracts ....................................... 57 4.2.2 Antibacterial activity of organic crude extracts ........................................ 58 4.2.3 Antibacterial activity of aqueous extracts at 1mg/ml................................ 59 4.2.4 Antibacterial activity of organic extracts at 1mg/ml ................................. 60 4.2.4 Antibacterial activity of organic extracts at 0.1mg/ml .............................. 61 4.3. Antibacterial activity of fractionated extracts ................................................. 62 4.3.1 Dead bark fractions at 100mg/ml .............................................................. 62 4.3.2 Live bark fractions at 100mg/ml ............................................................... 63 4.3.3 Leaf fractions at 100mg/ml ....................................................................... 64 4.3.4 Stem fractions at 100mg/ml ...................................................................... 65 4.4 Minimum inhibitory concentration (MIC) of fractions ................................... 66 4.41 Dead bark fractions at 10mg/ml ................................................................. 68 4.4.2 Leaf and stem fractions at 10mg/ml .......................................................... 69 4.4.3 Leaf and stem extract fractions in 1mg/ml................................................ 70 4.4.4 Leaf and stem extract fractions at 0.1mg/ml ............................................. 71 4.4.5 Leaf and stem extract fractions at 0.01mg/ml ........................................... 72 CHAPTER 5: DISCUSSION .................................................................................. 74 5.1. Antibacterial activity of crude extracts ........................................................... 74 5.1.1 T. sericea ................................................................................................... 74 5.2.2 C. imberbe ................................................................................................
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