IN-VTTRO SYNERG1STIC ANTIBACTERIAL EFFECT OF AQUEOUS AND ETHANOLIC EXTRACT OF GANAAA KOLA SEEDS AND G0NGRONAMIFA#B£»«N LEAVES ON SOME SELECTED ENTERIC PATHOGENS.

ABSTRACT

The in-vitro synergi1stic antibacterial effect of aqueous and ethanolic extract of Gongronema latifolium leaves and Garcinia kola seeds investigated against Siaphylococcus aureus, Pseudomonas aeruginosa and Escherichia coli. Agar well diffusion method was used for the antibacterial activity. The susceptibility of the test organisms to the plant extracts was performed and diameters of the inhibition were recorded. The ethanolic extract of the two plants in synergy showed the highest inhibitory activity. P. aeruginosa was the most susceptible (23.3mm) to the extract in synergy while S. aureus was inhibited the least (6.3mm) for the aqueous extract and was compared to standard control ciprofloxacin. The minimum inhibitory concentration (MIC) was evaluated for each plant extract and ranged from 6.25 to 100mg/ml. The result of this research work showed that the two plants in synergy had higher antibacterial activity compared to the plant when used individually. This can be employed as a form of antibacterial agent.

TABLE OF CONTENTS

TTTLEPAGE I

CERTIFICATION II

DEDICATION III

ACKNOWLEDGEMENT - IV

TABLE OF CONTENT V

LISTOF TABLES . VIIl

ABSTRACT IX

CHAPTER ONE

10 INTRODUCTION 1

1.1 AIM AND OBJECTIVES 2
CHAPTER TWO

2.0 LITERATURE REVIEW                                                                          . 3
2.0.1 Origin and distribution ofGarcinia kola 3
2.0.2 Origin and Distribution of G. latifolium 3

2.1 PHYTOCHEMISTRYANDPHYTOCHEMICALS 4

2.2 PHYTOCHEMICAL COMPONENTS 4

2.3 PHYTO CHEMICAL AND PHYSIOLOGICAL ABILITY OF Garcinia kola 6

2.4 PHYTOCHEMTCAL AND PHYSIOLOGICAL ABILITY OF

Gongronema lalifolium 6

2.5 MEDICINAL PLANTS 7

2.6 PHYTO CHEMICALS IN THE MEDICINAL PLANTS 8

2.7 SYNERC3STIC USE OF Gangronem  latifotium AND Garcmia kola AS DRUG       9

CHAPTER THREE

3.0 MATERIALS AND METHODS 11

3.1 SOURCE OF PLANTS MATERIAL 11

32 PREPARATION OF PLANT MATERIALS 11

33 PREPARATION OF AQUEOUS EXTRACT 11

3.4 PREPARATION OF ETHANOUC EXTRACT 11

3.5 SYNERGY TEST 12

3.5 SYNERGY TEST •'                   12

3.6 PYTOCHEMICAL SCREENING 12

3.7 QUALITATIVE ANALYSIS OF PHYTOCHEMICALS.

3.7.0 Test for tannins 12

3.7.1 Test For Saponins 12

3.7.2 Flavonoids 12

3.7.3 Steroids 13

3.7.4 Phenols 13

3.8 MICROSCOPY 14

r   . ,

3.9 GRAM STAINING 14

3.10 IDENTIFICATION OF BACTERIAL ISOLATES USING BIOCHEMICAL TESTS 15

3.10.0       Coagulase Test 15

3. 10.1        CatalaseTest 15

3. I0.2       Oxidasetest 15

3. 10.3      Urease Production Test      16

3. 10.4    Indole Test , 16

3.11 DETERMINATION OF MINIMUM INHIBITORY CONCENTRATION 16

3.12  TEST FOR ANTIMICROBIAL ACTIVITY. 17
CHAPTER FOUR

4.0 RESULTS. 18

4.1 ANTIMICROBIAL ACTIVITY OF EXTRACTS 18
CHAPTER FIVE

5.0 DISCUSSION 30

5.1 CONCLUSION 31
REFERENCES 32
Appendix

LIST OF TABLE

TABLE TITLE                                                                   PAGE

1 Preliminary (Qualitative) Phytochemical screening ofGarcinia kola

and Gongroneme     latifolium.        , 18

2 Aqueous extract of Gacmia kola against test isolates. 20

•/

3 Ethanolic extract of G. kola against test isolates 21

4 Aqueous extract of G. latifoftm against test isolates 22

5 Ethnolic extract of G. fatifoluim against test isolates 23
6     Synergistic effect of aqueous extract of both test plants. 24

7 Synergistic effect of Ethanolic extract of both test   plants. 25

8 Antimicrobial activity of extract from G. kola and G. latifolium, 26

singly and in combination against bacteria pathogens (diameter of inhibition zone (mm))

9 Minimum inhibitory concentration (mg/ml) of extract of G. kola and

G.   latiffolium singly and in combination against bacteria pathogens. 28

CHAPTER ONE

1.0 INTRODUCTION

Plants are important in our everyday existence. They produce the oxygen we breathe, serve as raw materials for many industrial products such as clothes, foot wears and many others. Plants also provide raw materials for our buildings and in the manufacture of biofuels, dyes, perfumes, pesticides. Plant act generally to stimulate and supplement the body’s healing forces (Fabiola et al., 2003). Natural infection and diseases are known to be treated with herbal remedies throughout the history of mankind. Infectious diseases are the number one cause of death accounting for approximately one half of all deaths in tropical countries (Iwu et al, 1999). Today plant materials continue to play a major role in primary health care  (Jonathan et al., 2007; Jonathan and Fasidi, 2005; Jonathan and Fasidi (2003). The different parts of  plant used include root, stem, flower, fruit, twin, exudates and modified plant organs (Mahesh and Satish, (2008) ; Uniyal et al., 2006). The use of plants in traditional medical practice has a long drawn history and remains the main stay of primary health care in most of the third world. Traditional  medicines are used by about 60% of the world population, in both developing and developed countries where modern medicines are predominantly used (Myithilypriya et al., 2007). While an estimated 60-80% African  population depend solely on herbal remedies for its primary health care needs. Some plants have been observed to be rich in secondary metabolites such as tannins, terponoids, alkaloids, flavonoids, phenols, steroids and volatile oil. These compounds are responsible for their therapeutic activities (Crown, 1999; Rabe and Vanstoden, 2000). Some plants have been used as anti microbial agents, especially their extracts either as decoctions, infusions or oral administration (Okemo et al., 2001).

1.1 AIM AND OBJECTIVES

AIM

The aim of the research project is to evaluate the antibacterial effect of combined extract of selected of G. latifolium leaves and G. kola seeds aganist enteric pathogens.

OBJECTIVES

1.) To determine the antibacterial effects of individual aqueous and ethanolic plant extracts.

  2. To evaluate and compare the antibacterial effect of combined extracts of G.    latifolium and G. kola extracts.

     3.      To determine the minimum inhibitory concentration of the combined extracts.

4.  To determine the minimum beteriocidal concentration of the combined extracts.

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