– Comparative Antibacterial, Anti-Venom And Phytochemical Studies Of Indigofera Capitata Kotschy And Indigofera Conferta Gillett In Albino Rat –
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ABSTRACT
This study was conducted to establish and compare anti-bacterial, anti-venom and phytochemical properties of Indigofera capitata and I. conferta in albino rats.
The two species of plants belong to the family Fabaceae. They are found growing on red soil orold farm land. The two plant materials were collected, dried, grounded and successively extracted using maceration with petroleum ether, ethanol and aqueous.
The extract of each solvent of the two plants was used for acute toxicity studies, in-vitro anti-bacterialand in-vivo anti-venom studies: Oedema forming activity, bleeding and clotting timeand histopathological examination of liver, kidney and heart were also carried out.
The most active extracts (ethanol and aqueous) were further subjected to thin layerchromatographic (TLC) studies. Antibacterial activities of all the extracts (petroleumether, ethanol and aqueous) of both plants showed significant effects (P<0.05) against the growth of Staphylococcus aureus and Salmonella typhi.
Indigofera confertapetroleum ether extract had MIC of 6.2 mg/ml and 12.5 mg/ml and MBC of 12.5 mg/mland 25.0 mg/ml, ethanol extract showed MIC of 12.5 mg/ml and MBC of 25.0 mg/mland aqueous extract showed MIC of 6.25
TABLE OF CONTENTS
Cover page: ……………….i
Title page:………………… ii
Declaration: ……………..…..iii
Certification: ………………… iv
Dedication: ………………….. v
Acknowledgements: …………..…….vi
Abstract: …………….….viii
Table of content: …………….x
List of Figures ………….xvii
List of Tables ………….xviii
List of Plates: ……………..xx
Appendix……………………….xxiii
CHAPTER ONE
1.0 Introduction………………..1
1.1 Plant Resources as Potential Traditional Medicine……………….1
1.2 Description and Distribution of Indigofera species…………….3
1.3 Staphylococci…………..…….3
1.4 Salmonella……………….4
1.5 Classification and morphology of snakes………….……5
1.6 General description of species of cobra…………….6
1.6.1 Taxonomy of cobra snakes…………..…….7
1.6.2 Species of cobra and their common names…………….7
1.7 Naja species (spitting cobras) Family Elapidae…….8
1.8 Anti- snake venom and its limitations……………10
1.9 Classification of anti-venom of snake……………11
1.10 Statement of the research problems……..….11
1.11 Justification of the studies………….12
1.12 Aim and Objectives……………….13
1.12.1 Aim……………13
1.12.2 Objectives…….……..13
1.13 Hypothesis………..….13
CHAPTER TWO
2.0 Literature review…………15
2.1 Morphological Features and Classification of Indigofera capitata and Indigofera conferta………15
2.2 Phytochemical Constituents of Indigofera species………..15
2.3 Biological Activities of some Indigofera species……..…16
2.4 Indigofera Species used against Bacteria………..18
2.5 Traditional uses of Some Indigofera species………19
2.6 Medicinal uses of some Indigofera species…………20
2.7 Plant Constituents as Crude Drugs…….…….22
2.8 Ethnomedical Uses Of Some Indigofera Species……..23
2.9 Evaluation of Plant Crude Drugs…………..23
2.10 Flavonoids and their Classification………….25
2.10.1 Medicinal uses of flavonoid….………..27
2.10.2 Toxicity of flavonoids…………….27
2.11 Saponins…………….28
2.12 Snake Venoms…………….29
2.12.1Composition of Snake Venom………….29
2.12.2 Types Of Snake Venom………….30
2.12.3 Therapeutic role of Snake Venom……………..30
2.12.4 Fibrinolytic Activity of Snake Venom……………..30
2.12.5 Cardiotonic Activity of Snake Venom……….31
2.12.6 Muscular Depolarization and Haemolysis Of Snake Venom………..31
2.13 Naja nigricollis Venom and its effect to Man…..……..31
2.14 Bleeding and Clotting Time…………..32
2.15 Oedema Forming Activity…………….32
2.16 Medicinal Plants Active Against Snake Bites………….33
CHAPTER THREE
3.0 Materials and Methods…………………49
3.1 Plant Collection and Identification…………49
3.2 Preparation of Plant Materials…………..49
3.3 Extraction of Indigofera capitata and Indigofera conferta
whole plants………..………..……………..……..…..……………………….49
3.4 Experimental animals………….……….………..……….………………..50
3.5 Antibacterial Activities of Crude Extracts of Indigofera capitata and Indigofera conferta……..50
3.6 Acute Toxicity Studies of Indigofera capitata and Indigofera confert……..51
3.7 In vivo Evaluation of the Crude Extracts of Indigofera capitata and Indigofera conferta against Naja nigricollis envenomed Rats.…….51
3.7.1 Capturing of snake and collection of venom…………………………..…….51
3.7.2 Determination of minimum lethal dose of Naja nigricollis venom (LD99) on mice…………… 52
3.7.3 Determinations of bleeding and clotting time of rats treated with different extracts of Indigofera capitata and Indigofera
conferta against Naja nigricollis envenomed rats…..52
3.7.4 Determination of Oedema forming activity on the effect of different extracts of Indigofera capitata and Indigofera conferta against Naja nigericollis envenomed rats………… 54
3.8 Histopathological studies on the effect of extracts of Indigofera capitata and Indigofera conferta against Naja
nigricollis envenomed Rats…..……..55
3.9 Thin Layer Chromatography of the most Active Extracts of Indigofera capitata and Indigofera conferta.…………56
3.10 Statistical Analysis………….……………………………………………….57
CHAPTER FOUR
4.0 Results…………………….58
4.1 Extractive yield of Indigofera capitata and Indigofera conferta…..…………………59
4.2 Acute Toxicity Studies of Indigofera capitata and Indigofera conferta in Albino Rats………..………60
4.3 Antibacterial Activities of different Extract Indigofera capitata and I. conferta……………………..62
4.4 Bleeding and Clothing Time of Albino Rats Treated with different Extracts of Indigofera capitata against LD Naja nigricollis venom…….…………..70
4.5 Bleeding and Clotting Time of Albino Rats Treated different Extracts of Indigofera conferta against Naja nigricollis Venom…….………..74
4.6 Oedema Forming Activities in Albino Rats Treated with different Extracts of Indigofera capitata against LD Naja nigricollis venom……………..….. 78
4.7 Oedema Forming Activities in Albino Rats Treated with different Extracts of Indigofera conferta against LD99
Naja nigricollis Venom……………..….…82
4.8 Histopathological Investigation of the Effect of Different Extracts of Indigofera capitata and Indgofera conferta against Naja nigricollis envenomed Rats…….………… 86
4.8.1 Histopathological responses of liver, kidney and heart in albino rats treated with different doses of petroleum ether
whole plant extracts of Indigofera capitata against LD99 Naja nigricollis enom……………….93
4.8.2 Histopathological responses of liver, kidney and heart inalbino rats treated with different doses of ethanol whole plant extracts of Indigofera capitata against LD99 Naja nigricollis venom…….…………97
4.8.3 Histopathological responses of liver, kidney and heartin albino rats treated with different doses of aqueous whole
plant extract of Indigofera capitata against LD99 Naja nigricollis venom……..…………101
4.8.4 Histopathological responses of liver, kidney and heart in albino rats treated with different doses of petroleum ether whole plant extract of Indigofera conferta against Naja nigricollis venom……….…………….…..….105
4.8.5 Histopathological responses of liver, kidney and heart inalbino rats treated with different doses of ethanol whole plant extract of Indigofera conferta against LD99 Naja nigricollis venom…………..109
4.8.6 Histopathological responses of liver, kidney and heartin albino rats treated with different doses of aqueous whole plant of Indigofera conferta against LD99 Naja nigricollis venom……………….…….113
4.9 Thin Layer Chromatography of the most Active Extracts of Indigofera capitata and Indigofera conferta………..…………..……….117
CHAPTER FIVE
5.0 Discussion…………………118
CHAPTER SIX
6.0 Summary, Conclusion and Recommendation…………………….……131
6.1 Summary……………….131
6.2 Conclusion…………….132
6.3 Recommendation…………….134
References……………………135
Appendices………………147
REFERENCES
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