TITLE PAGE

EVALUATION OF ANTIBACTERIAL ACTIVITIES OF Bryophyllum pinnatum AND

Ocimum gratissimum LEAF EXTRACTS AGAINST Selected Gram-positive and Gram-
negative bacteria

BY

IFEZUE, KOSISO JENNIFER

REGISTRATION NUMBER: NAU/2019484055

A PROJECT SUBMITTED TO THE DEPARTMENT OF APPLIED MICROBIOLOGY

AND BREWING
FACULTY OF BIOSCIENCES
NNAMDI AZIKIWE UNIVERSITY, AWKA.

IN PARTIAL FULFILMENT OF THE REQUIREMENTS FOR THE AWARD OF THE

DEGREE OF BACHELOR OF SCIENCE (B.Sc.) IN APPLIED MICROBIOLOGY AND
BREWING.

SUPERVISOR: MISS ABANA, C. C.

JANUARY, 2024.

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 CERTIFICATION

I certify that this project titled “Evaluation of antibacterial activities of Bryophyllum pinnatum
and Ocimum gratissimum extracts against selected Gram-positive and Gram-negative bacteria
was carried out by IFEZUE, KOSISO JENNIFER with the registration number
NAU/2019484055 has satisfactorily completed the requirements for the award of Bachelor of
Science (B. Sc.) degree in Applied Microbiology and Brewing, Nnamdi Azikiwe University
Awka, Anambra State.

IFEZUE, KOSISO JENNIFER Date

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 APPROVAL

We, the undersigned hereby approve that this project titled “EVALUATION OF
ANTIBACTERIAL ACTIVITIES OF Bryophyllum pinnatum AND Ocimum gratissimum
EXTRACT AGAINST SELECTED GRAM-POSTIVE AND GRAM-NEGATIVE BACTERIA”
presented by IFEZUE, KOSISO JENNIFER with the registration number NAU/2019484055
fulfilled the requirements and should be accepted for the reward of Bachelor of Science (B. Sc.),
Nnamdi Azikiwe University Awka, Anambra State.

Miss Abana, C. C. Date

(Supervisor)

Prof. S. O. Umeh Date

(Head of Department)

Prof. I. O. Igwilo Date

(Dean, Faculty of Biosciences)

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External Examiner Date

DEDICATION

I dedicate this research project to my Heavenly Father, for his abundant mercy, grace, and loving
kindness in my life throughout my undergraduate program and the completion of this work.

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 ACKNOWLEDGEMENTS

I acknowledge God Almighty for his guidance, protections, love, and good health throughout the
course of this study.

My immense gratitude goes to my supervisor, Miss C. C. Abana for her advice, support, and
guidance through this my research. I am humbled to have worked under our tutelage. Thank you
so much for your time, effort, and untiring commitment to ensure the positive outcome of this
work.

My sincere appreciation goes to our Head of Department, Prof S.O. Umeh for her motherly
carriage in running the affairs of our department. I also appreciate my lecturers for the
knowledge impacted on me and for the advice and efforts towards my academics.

I thankfully acknowledge my parents, Mr. and Mrs. Boniface Ifezue for their underlying love,
prayers, moral and financial support throughout this period.

I am also grateful to Fr. Eya Thaddeus and Fr. Obiesie Peter for their support and advice during
this work and to my friends, Ifeoma, Stephanie, Chinecherem and Chidera for their help during
this work.

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 ABSTRACT

The misuse of antimicrobials in humans, and animals are the main drivers in the development of
drug-resistant pathogens. Antimicrobial resistance (AMR) makes infections harder to treat and
other medical procedures, like surgery, caesarean sections, and cancer chemotherapy difficult.
Medicinal plants are those which, introduced in a human or animal organism, produce
pharmacological activity. Bryophyllum pinnatum is a succulent plant from the Crassulaceae
family with abundant application in traditional medicine, used to treat different kinds of
ailments. It is commonly called “miracle plant” and “odaa opue” in Igbo. Ocimum gratissimum is
a perennial herbaceous plant with very strong aromatic smell from the family of Lamiaceae with
medicinal properties. It is commonly called “scent leaf”. The effect of these two leaves against
some selected bacteria (Staphylococcus aureus, Bacillus species, Enterobacter species, and
Escherichia coli) was analyzed using ethanol and sterile water. The leaves of B. pinnatum and O.
gratissimum were obtained from Aroma and Ifite-UNIZIK in Awka Local Government Area,
Anambra State. Using the methodology described by …………… the leaves and their extracts
were carefully prepared using ethanol and water as solvents. The antimicrobial activity of the
plants was assessed using the agar well diffusion method as described by Amala et al. (2021).
The ethanol extract of B. pinnatum showed the best antibacterial effect on Escherichia coli with
the highest inhibition zone being 20mm while it had no effect on Enterobacter species. The
synergistic activity of ethanol extract of both leaves was most effective on E. coli, followed by
Bacillus species then Enterobacter species. The synergistic activity of aqueous extract of both
leaves showed no effect on other isolates except Bacillus species with zone of inhibition
measured 7mm. From the findings of this study, it can be concluded that ethanol extracts of the
leaves were more effective than the aqueous extract and that the synergistic effect of the
ethanolic extract was more effective than the singly used extracts. Bryophyllum pinnatum and
Ocimum gratissimum inhold various medicinal properties that can be used in treatment of
various illness, hence, can be incorporated in drug production.

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 TABLE OF CONTENTS

Title page i

Certification ii

Approval iii

Dedication iv

Acknowledgements v

Abstract vi

Table of contents vii

List of figures x

List of tables xi

CHAPTER ONE

1.0 INTRODUCTION 1

1.1 The Aim of Research 3

1.2 Specific Objectives 3

1.3 Significance of The Study 3

CHAPTER TWO

2.0 LITERATURE REVIEW 4

2.1 Origin of medicinal plants 4

2.2 Antimicrobial activities of medicinal plants, a possible alternative in

treatment of multidrug resistant microbes. 5

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2.3 Bryophyllum pinnatum plant 6

2.4 Ocimum gratissimum plant 10

2.5 Synergistic activities of both plants 13

2.6 Test organisms 13

2.6.1 Escherichia coli 13

2.6.2 Staphylococcus aureus 14

2.6.3 Bacillus species 15

2.6.4 Enterobacter species 16

CHAPTER THREE

3.0 MATERIALS AND METHODS 18

3.1 Study area 18

3.2 Study design 18

3.3 Collection and authentication of leaves sample 18

3.4 Sample preparation 18

3.5 Extraction of leaves 18

3.6 Media Preparation 19

3.7 Sources and collection of bacteria isolates. 19

3.8 Confirmation and standardization of test organisms 19

3.9 Identification of the Test Organism 19

3.9.1 Using gram staining. 19

3.9.2 Using biochemical test. 20

3.10 Preparation of test organisms and seeding of Mueller Hinton agar plates 20

CHAPTER FOUR

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Results 21

CHAPTER FIVE

Discussion 27

Conclusion 28

References 29

Appendix 34

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 LIST OF FIGURES

Figure 1: Pictorial representation of Bryophyllum pinnatum plant. 9

Figure 2: Pictorial representation of Ocimum gratissimum plant. 12

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 LIST OF TABLES

Table 1: Diameter of zone of inhibition of ethanol extract of Bryophyllum pinnatum

and Ocimum gratissimum 22

Table 2: Diameter of zone of inhibition of aqueous extract of Bryophyllum pinnatum

and Ocimum gratissimum 23

Table 3: zone of inhibition of the synergistic effect of ethanol extract of both

Bryophyllum pinnatum and Ocimum gratissimum 24

Table 4: zone of inhibition of the synergistic effect of aqueous extract of both

Bryophyllum pinnatum and Ocimum gratissimum 25

Table 5: Diameter of zone of inhibition of ampicillin against the test organisms. 26

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