PHYTOCHEMICALS AND ANTIMICROBIAL ACTIVITIES OF NEWBOULDIA LEAVIS ON E.COLI AND STAPHYLOCOCCUS AUREUS

ABSTRACT

The plant kingdom offers a variety of a wide range of plants with natural antimicrobial and medicinal values. The widespread of diseases of microbial origin remains alarming, particularly of serious concerns in the tropics and sub-tropics. The high costs of newer and effective drugs and other factors have made the choice of traditional herbal medicine considered against these pathogenic microbes, and making it more economical. N. leavis is one of the plants with magical effects, which is commonly called tree of life. N. leavis has been reported to have medicinal value ranging from anti-inflammatory, anti-microbial, anti-fungi, analgesic and wound healing properties.  The results obtained from the phytochemical analysis of N. leavis leave extracts showed the presence of Alkaloids, Phenols, Tannins, Protein etc. The highest yield was obtained from the water extract with protein giving a yield of 2.92. Phytochemical are naturally occurring and are believed to be effective in combating or preventing disease due to their antioxidant properties. While in the course of this study, it was discovered that Alkaloid, Tannins, Ether, Protein, Saponin and Phenol were all positive. While in the antimicrobial effect of N. leavis it was discovered that the highest yield was obtained from the water extract with the zone of inhibition on E.coli was 38mm. It was discovered that N. leavis has a clear zone of inhibition on E.coli and meanwhile there were no zone of inhibition on Methicillin resistant S. aureus in recommendation, I suggest that N. leavis is used to treat and E.coli induced infection.

TABLE OF CONTENTS

Title Page                                                                                                                                i

Certification                                                                                                                            ii

Declaration                                                                                                                              iii

Dedication                                                                                                                              iv

Acknowledgements                                                                                                                v

List of Tables                                                                                                                          ix

Abstract                                                                                                                                  x

CHAPTER ONE

1.1       Introduction                                                                                                                1

1.2       Medicinal Uses of N. leavis                                                                                       2

1.3       Aims and Objectives                                                                                                   3

1.4       Objectives                                                                                                                   3

CHAPTER TWO

2.1       Background of the Study                                                                                           4

2.2       Newbouldia leavis                                                                                                      5

2.2.1    Scientific classification of Newbouldia leavis                                                5

2.2.2    Medicinal uses of Newbouldia leavis                                                                          6

2.3       Phytochemicals                                                                                                          7

2.3.1    Phytochemical constituents of plants                                                                        8

2.4       Terpenoids                                                                                                                  8

2.5       Saponins                                                                                                                     9

2.6       Tannins                                                                                                                       10

2.7       Alkaloids                                                                                                                    11

2.8       Escherichia coli                                                                                                           12

2.8.1    Taxonomy                                                                                                                   12

2.8.2   Morphology and Biochemical Characteristics                                                             13

2.8.3   Habitat and Growth Characteristics                                                                            13

2.8.4    Pathogenic Escherichia coli                                                                                        14

2.8.5   Gastrointestinal Infection                                                                                            14

2.9       Staphylococcus aureus                                                                                                14

2.9.1   Taxonomy                                                                                                                    15

2.9.2   Morphology and Growth Characteristics                                                                    15

2.9.3    Natural Habitat                                                                                                           17

2.9.4    Virulence factors of S. aureus                                                                                    18

2.9.5   Pathogenesis of S. aureus                                                                                            18

CHAPTER THREE: MATERIALS AND METHODS                                                 

3.1 Collection and Preparation of Plant Material                                                                   19

3.2  Preparation of Plant Extract                                                                                            19

3.3  Extraction of Plant Material                                                                                            19

3.4  Phytochemcial Testing                                                                                                     20

3.4.1 Determination of Saponin                                                                                              20

3.4.2 Determination of Total Phenol                                                                                      21

3.4.3 Protein Determination                                                                                                    22

3.4.4 Test for Tannins                                                                                                             23

3.4.5 Test for Alkaloids                                                                                                          23

3.5 Media Used                                                                                                                      24 

3.5.1 Preparation of Media                                                                                                     24

3.5.2 Sterilization                                                                                                                    24

3.5.3 Sources of the micro-organisms Used                                                                           24

3.5.4  Biochemical confirmation at the test organism                                                             25

3.6       Identification of bacteria isolates                                                                               25

3.6.1    Gram staining                                                                                                              25

3.7       Biochemical cultural confirmation                                                                              26

3.7.1 Catalase Test                                                                                                                  26

3.7.2 Coagulase Test                                                                                                               26

3.7.3    Citrate Test                                                                                                                 26

3.7.4    Motility, Indole, Urease Test (MIU)                                                                          27

3.7.5 Triple Sugar iron Test                                                                                                     27

3.7.6 Oxidase Test                                                                                                                  28

3.8 Antimicrobial Susceptibility Testing                                                                                 28

3.8.1    Preparation of Paper Disc                                                                                           28

3.8.2 Agar Disc Diffusion Method                                                                                         28

CHAPTER FOUR: RESULTS                                                                                           30

4.1       The Morphology and biochemical characterization of isolates used in this study      29

4.2       The qualitative phytochemical composition of Newbouldia leavis from

different extracts                                                                                                        29

4.3       Antimicrobial activity of water extract of Newbouldia leavis against the bacterial

isolate measured by zone of inhibition                                                                       29

4.4       Antimicrobial activity of ethanol extract of Newbouldia leavis measured by

zone of inhibition                                                                                                        29

CHAPTER FIVE: DISCUSSION, CONCLUSION AND RECOMMENDATIONS 

5.1       Discussion                                                                                                                   34

5.2       Conclusion                                                                                                                  35

5.3       Recommendations                                                                                                      36

REFERENCES

LIST OF TABLES

TABLE                                              TITLE                                                            PAGE

4.1:           Morphology and Biochemical Identification of Isolate                               30

4.2:         The Qualitative Phytochemical Composition of Newbouldia leavis

      from different extracts                                                                                     31

4.3:        Antimicrobial Activity of Water Extract of Newbouldia leavis

 measured by Zone of Inhibition                                                                          32                         

4.4:      Antimicrobial Activity of Ethanol Extract of Newbouldia leavis

 measured by Zone of Inhibition                                                                          33

CHAPTER ONE

1.1       Introduction

The wide spread of diseases of microbial origin remains alarming, very high and particular of serious concerns in the tropics and sub-tropics. Microbial infections can lead to serious and lethal complications. The high cost of newer and effective drugs and other factors, has made the choice of traditional herbal remedies against these infections unavoidable and more economical (Okonko et al., 2009). Nosocomial infection is an infection whose development is favored by hospital environment. It may develop in a hospitalized patient without having been present or incubating at the time of admission, or it may be acquired in hospital but only appears after discharge. Since ancient times, plants have been found to be an essential source of natural products for maintaining human health. There are numerous compounds in plants that have efficient effects on animals and posses’ high therapeutic properties, which can be utilized in the treatment and care of human and other animal diseases (Ubulom et al., 2013). Modern day medicine at the turn of the millennium has been characterized by breath-taking advances that are revolutionizing the practice of medicine. However, invasive devices especially intravascular devices of all types and urinary catheters continue to be essential for the management of critically ill patients. Permitting lifesaving treatment and physiologic monitoring, these ubiquitous devices, unfortunately, are a major cause of institutionally acquired infections, especially in intensive care units. These infections have fueled the advancing crisis in antibiotic resistance and have major economic consequences in the form of extended length of hospital stay and attributable mortality. The risk of device-related infection, more than anything else, has forced medicine to accept the necessity for nosocomial infection control. In general, invasive devices of all types are far more important in determining susceptibility to nosocomial infection than are underlying diseases. The emergence and spread of microbes that are resistant to cheap and effective first-choice drugs has become a common occurrence. The problem is even more evident in bacterial infections which contribute most to the global infectious disease burden such as diarrheal, respiratory tract, meningitis, sexually transmitted infections, and tuberculosis (WHO, 2002). Plants have traditionally provided a source of hope for novel drug compounds, as plant herbal mixtures have made large contributions to human health and well-being. Owing to their popular use as remedies for many infectious diseases, searches for substances with antimicrobial activity in plants are frequent (Betoni et. al., 2006). Plants are rich in a wide variety of secondary metabolites, such as Tannins, Terpenoids, Alkaloids, and Flavonoids, which have been found in vitro to have antimicrobial properties (Lewis  et. al., 2006).

1.2       Medicinal Uses of N. laevis

Scientifically, N. leavis has been reported to have medicinal value ranging from anti inflammatory, antioxidant, anti-microbial, anti-fungi, analgestic and wound healing properties (Chukwujeku et al., 2005; Kuete et aI., 2007). Specifically, the stem bark mixed with clay and red pepper has been reported to be effective against pneumonia, fever, cold, cough and for treating different illness like bone lesions (Idu, 2009). N. leavis is widely used in African folk medicine for the treatment of malaria and fever, stomachache, coughs, sexually transmitted diseases, tooth ache, breast cancer, and constipation (Arbonnier et al., 2004).

In South Eastern and part of the Midwestern Nigeria, the plant is used for the treatment of septic wounds and eye problems according to (Usman et al., 2007). Scientific reports on the phytochemical constituents of the plant revealed the presence of alkaloids and phenylpropanoids in the root, flavonoids, and tannins in the leaf as revealed by (Chukwujeku et al., 2005).

1.3       Aims and Objectives 

To investigate the phytochemical and antimicrobial activity of Newbouldia leavis on Methicillline resistant Staphylococcus aureus and E.coli.

1.4       Objectives

1.         To determine the qualitative phytochemical constituents of Newbouldia leavis.

2.         To isolate and confirm the text organisms.

3.         To determine the antimicrobial activity of Newbouldia leavis on Methicillin resistant Staphylococcus aureus and E.coli.

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