PHARMACOGNOSTIC AND ANTIBACTERIAL STUDIES OF ACACIA SIEBERIANA VAR WOODII (FABACEAE) STEM BARK

TABLE OF CONTENTS
Title Page
Abstract
Table of content
Abbreviations, Symbols and Glossaries

CHAPTER ONE
1.0       INTRODUCTION
1.1       Traditional medicine
1.2       Plants in traditional medicine
1.3       Medicinal plants as antibacterial agents
1.4       Statement of Research Problem
1.5       Justification
1.6       Aims and Objectives
1.7       Hypothesis

CHAPTER TWO
2.0       LITERATURE REVIEW
2.1       Botanical description of fabaceae
2.1.1    Economic and medicinal importance of fabaceae
2.1.2    Reported phytochemical constituents of fabaceae
2.2       Botanical description of genus Acacia
2.2.1    Economic importance of Acacia species
2.2.2  Reported Phytochemical Constituents of Acacia species
2.3       Botanical description of Acacia sieberiana
2.3.1    Economic and medicinal importance of Acacia sieberiana
2.3.2    Reported phytochemical constituents of Acacia sieberiana
2.3                   Enteric Bacteria

CHAPTER THREE
3.0       MATERIALS AND METHOD
3.1       Materials, Solvents and Reagents
3.1.1    Materials
3.1.2    Solvents
3.1.3    Reagents
3.2       Collection and Identification of A. sieberiana
3.3       Preparation of A. sieberiana
3.4       Determination of Botanical Features and Physical Constants A. sieberiana Stem Bark
3.4.1    Microscopical studies of the stem bark
3.4.2    Chemomicroscopical studies of the stem bark
3.4.3    Physical constants determination of the stem bark
3.5       Screening of Phytochemical Constituents of A. sieberiana Stem Bark
3.5.1    Extraction of the plant stem bark
3.5.2    Preliminary phytochemical studies of the hexane and methanol extracts of the stem bark
3.5.3    Thin layer chromatography of the hexane extract
3.5.4    Column chromatography of the hexane extract
3.6       Determination  of  Margin  of  Safety  of  A.  sieberiana  Stem  Bark Extracts
3.6.1    Acute toxicity (LD50)
3.7       Determination of Antibacterial Studies of A. sieberiana Stem Bark against Some Enteric Bacteria
3.7.1    Bacterial isolates
3.7.2    Preparation of stock solution
3.7.3    Preparation of media for bacterial growth
3.7.4    Mc-Farland 0.5 barium sulphate turbidity standard
3.7.5    Determination of zones of inhibition for Fraction AS1, hexane and methanol extracts from A. sieberiana
3.7.6    Determination of minimum inhibitory concentration of Fraction AS1, hexane and methanol extracts from A. sieberiana
3.7.7    Determination of minimum bactericidal concentration of Fraction AS1, hexane and methanol extracts from A. sieberiana

CHAPTER FOUR
4.0       RESULTS
4.1       Determination of Botanical Features and Physical Constants of A. sieberiana Stem Bark
4.1.1    Microscopical Studies of the Stem Bark
4.1.2    Chemo microscopical Studies of the Stem Bark
4.1.3    Physical Constants Determination of the Stem Bark
4.2       Screening of Phytochemical Constituents of A. sieberiana Stem Bark
4.2.1    Extraction of the stem bark
4.2.2    Preliminary phytochemical studies of the stem bark
4.2.3    Thin-Layer Chromatographic studies of hexane extract from  A. sieberiana
4.2.4    Column chromatography of hexane extract from A. sieberiana
4.3       Margin of Safety of A. sieberiana Stem Bark Extracts
4.3.1    Acute toxicity study (LD50) of the stem bark extracts
4.4       Determination of Antibacterial Studies of A. sieberiana Stem Bark Extracts against Some Enteric Bacteria
4.4.1    Zones of inhibition of Fraction AS1, hexane and methanol extracts from A. sieberiana against some enteric bacteria
4.4.2    Minimum inhibitory concentration of Fraction AS1, hexane and methanol extract from A. sieberiana against some enteric bacteria
4.4.4    Minimum bactericidal concentration of Fraction AS1, hexane and methanol extract from A. sieberiana against some enteric bacteria

CHAPTER FIVE
5.0       DISCUSSION

CHAPTER SIX
6.0       SUMMARY, CONCLUSION AND RECOMMENDATION
REFERENCES
APPENDICES

ABSTRACT
Acacia sieberiana var. Woodii (Fabaceae) is traditionally used as a remedy for stomach aches and ulcers among some localities in Northern Nigeria. The plant was investigated sequel to the reports of its ethnomedicine uses in the management of stomach ache and ulcers. Anatomical and physical constants studies were carried out using standard procedures. Phytochemical (preliminary studies, thin layer and column chromatography) were also carried out on the hexane and methanol extracts. Toxicity and antibacterial studies of the hexane and methanol extracts, and the isolated compound were investigated. The microscopical studies revealed the presence of cellulose cell wall, lignified and suberized cell walls, tannins and calcium crystals. Anatomical features identified in the stem bark consist of epidermis, phellogen, phelloderm, and cortex. The percentage moisture content and ash values were observed to be 9.0% and 10.5% respectively. Water extractive value was 1.2% while ethanol extractive value was 1.6%.The preliminary phytochemical studies as well as the TLC chromatogram of the hexane extract revealed the presence of steroids and triterpenes while the methanol extract contains tannins, flavonoids, alkaloids, steroids and triterpenes. Fraction AS1collected through column chromatography was suggested to be a hydrocarbon on the basis of its colourless and oily nature. The acute toxicity study carried out indicated that the hexane extract was slightly toxic while the methanol extract was moderately toxic. Antibacterial studies of the stem bark carried out showed zones of inhibition between 16 to 27 mm for the hexane and methanol extracts against Helicobacter pylori, Escherichia coli, Salmonella typhi and

Shigella dysenteriae. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were between the range of 2.5 and 10 mg/ml and 5.0 to 20 mg/ml respectively. Fraction AS1 exerted a dimension of zones of inhibition between 26 to 30 mm against all the bacterial strains except S. typhi. It was also able to exert MIC and MBC against E. coli and H. pylori at 12.5 and 25µg/ml respectively, while it had MIC and MBC at the lowest concentrations of 6.25 and 12.5 µg/ml against S. dysenteriae. Results of the present studies had shown that A. sieberiana and fraction AS1 collected from this plant have wide antibacterial property.


CHAPTER ONE
1.0              INTRODUCTION
1.1              Traditional Medicine
Traditional medicine can be said to be a comprehensive knowledge system that encompasses the utilization of substances, dosages and practices based on socio-cultural norms and religious beliefs as well as witnessed experiences and observations of a specific group (Tom et al., 2008). The practice is used in various therapies by the indigenous population all over the world. It has been documented that about 80% of the people in developing countries rely on traditional medicines for their primary health care need, which can be attributed to increased poverty, ignorance as well as unavailability of modern health facilities (Omonike, 2010; Christiana et al., 2012).

1.2              Plants in Traditional Medicine
Plant based traditional remedies are the oldest forms of health care delivery known to mankind and it has been used in all cultures throughout history. Plants have been used as early as 5000-4000BC in different parts of the world for the treatment of various ailments (Nyananyo and Akada, 2011; Kunle et al., 2012). A vast knowledge on how to use these plants against diseases may be expected to have accumulated in areas where the use of these plants is of great importance. The knowledge of these plants was developed gradually as it passed from generation to generation and has laid foundation of many health care systems all over the world (Azmatullah et al., 2011; Kunle et al., 2012).......

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