ABSTRACT
The study evaluated the antibacterial activity of sweet orange (Citrus sinensis) and lime (Ctrus aurantifolia) leaf extract with different solvents (methanol and aqueous) on some pathogenic organisms such as Pseudomonas aeruginosa, Staphylococcus aureus and Salmonella typhi. The fresh leaves of lime (Citrus aurantifolia) and orange (Citrus sinensis) samples were collected at National Root Crop Research Institute Umudike in Ikwuano, Abia State, Nigeria. The result showed that the Methanol and aqueous leaf extracts of lime (Citrus aurantifolia) and Sweet orange (Citrus sinensis) inhibited the growth of the entire bacterial isolates tested. It was observed that the methanol extracts had more significant inhibitory effect on the test organisms than the aqueous extracts. The methanolic leaf extracts of lime (Citrus aurantifolia) recorded a pronounced antibacterial activity against the test pathogens with zones of inhibition varying between 20 mm against Staphylococcus aureus isolate to 23 mm against Pseudomonas aeruginosa then 22mm against Salmonella typhi at the concentrations of 200mg/ml, whereas the aqueous extracts of lime (Citrus aurantifolia) showed minimal antibacterial activity against the isolates with zones of inhibition varied between 18 mm against Staphylococcus aureus isolate to 21 mm against Pseudomonas aeruginosa then 20mm against Salmonella typhi at the concentrations of 200mg/ml compared to the methanol extracts. The methanolic leaf extract of Sweet orange (Citrus sinensis) was more effective in inhibiting Salmonella typhi with zone of inhibition ranging between 15mm and 21mm as compared to Pseudomonas aeruginosa with zone of inhibition ranging between 13mm to 20mm then Staphylococcus aureus with zone of inhibition ranging between 13mm to 18mm. The antimicrobial activity of aqueous leaf extracts of Sweet orange (Citrus sinensis) was evaluated against the test organisms and it was more effective in inhibiting Pseudomonas aeruginosa with zone of inhibition ranging between 14mm to 19mm, this is followed by Salmonella typhi with inhibition zone ranging from 14mm to 18mm whereas Staphylococcus aureus had the list inhibition zone ranging from 14mm to 16mm. The results of this study suggest that the leaf extract of sweet orange (Citrus sinensis) and lime (Ctrus aurantifolia) can be used as an antibacterial agent against infections caused by Pseudomonas aeruginosa, Staphylococcus aureus and Salmonella typhi. It could be concludeD that methanol was the best extractive solvent for antibacterial activity of sweet orange (Citrus sinensis) and lime (Ctrus aurantifolia) against all the tested organisms.
TABLE OF CONTENTS
Title
Page i
Certification ii
Dedication iii
Acknowledgement iv
Table
of Contents v
List
of Tables vii
Abstract viii
CHAPTER ONE
1.0 Introduction 1
1.1 Aim
and Objectives 3
CHAPTER TWO
2.0 Literature Review 4
2.1 Plant Description 5
2.2 Brief
History and Geographical Distribution of C. aurantifolia 6
2.3 Botany of Citrus sinensis 6
2.4 Domestic Uses of Citrus aurantifolia 7
2.5 Ethnomedical Uses of Citrus aurantifolia 8
2.5.1 Leaves 8
2.5.2 Juice 9
2.5.3 Other parts of the plant 9
2.6 Pharmacological Activities of C.
aurantifolia and C sinensis 10
2.6.1 Antibacterial activity 10
2.6.2 Antifungal and antiaflatoxigenic activity 10
2.6.3 Anticancer/ cytotoxic activity 11
2.6.4 Antioxidant activity 11
2.6.5 Immuno-modulatory activity 12
2.6.6 Nutritional content of C. aurantifolia and C.
sinensis fruit juice 12
2.7 Phytomedicine 14
2.8 Risk
of Phytomedicine 14
2.9 Phytochemicals 14
2.10 Classes
of Phytochemical 15
2.10.1 Alkaloids 15
2.10.2 Glycosides 16
2.10.3 Flavonoids 16
2.10.4 Phenolics 17
2.10.5 Saponins 17
2.10.6 Tannins 18
2.11 Mechanism
of Action of Phytochemical 18
2.12 Health Benefits of Phytochemicals Obtained
from Citrus 19
2.13 Test Organisms 20
2.13.1 Salmonella
typhi 20
2.13.2 Staphylococcus aureus 21
2.13.3 Pseudomonas aeruginosa 21
CHAPTER THREE
3.0 Materials
and Methods 23
3.1 Study
Area 23
3.1 Sample
Collection 23
3.2 Sterilization
Methods 23
3.3 Preparation
of Crude Extracts 23
3.4 Bacterial
Strains and Cultural Conditions 24
3.5 Confirmation
of Test Isolate 24
3.5.1 Morphological
appearance 24
3.5.2 Gram Staining 25
3.5.3 Motility test 25
3.5.4 Biochemical
Tests 25
3.6 Evaluation of Antimicrobial Activity 28
3.7 Determination
of the Minimum Inhibitory Concentration (MIC) 28
3.8 Determination of the
Minimum Bactericidal Concentration (MBC) 29
CHAPTER FOUR
4.0 Result 30
CHAPTER FIVE
5.0 Discussion,
Conclusion and Recommendation 37
5.1 Discussion 37
5.2 Conclusion 40
5.3 Recommendation 40
References 41
LIST
OF TABLES
|
TABLE
|
TITLE
|
PAGE
|
|
1
|
Morphological and Biochemical Identification, Gram
Reaction and Sugar Utilization Profile of the
test organisms
|
31
|
|
2
|
Antimicrobial activity of the Methanol and aqueous leaf
extracts of lime (Citrus aurantifolia)
on the test organisms
|
32
|
|
3
|
Antimicrobial activity of the Methanol and aqueous leaf
extracts of Sweet orange (Citrus
sinensis) on the test organisms
|
33
|
|
4
|
The antimicrobial activities of the control drug
(Streptomycin)
|
34
|
|
5
|
The MIC and MBC values (mg/ml) of the Methanol and
aqueous leaf extracts of lime (Citrus
aurantifolia) on the test organisms
|
35
|
|
6
|
The MIC and MBC values (mg/ml) of the Methanol and
aqueous leaf extracts of Sweet orange (Citrus
sinensis) on the test organisms
|
36
|
CHAPTER ONE
1.0 INTRODUCTION
Medicinal
plants are the natural resources in developing of new drugs. Nature has been a
source of medicinal agent for thousands of years and an impressive number of
modern drugs have been isolated from natural sources, based on their use in traditional
medicine (Chaudhari et al., 2012).
This is due to increased source awareness of the limited ability of the
synthetic pharmaceutical products to control major diseases. The basic
molecular and active structures for synthetic fields are provided by rich
natural sources (Preeti et al.,
2010). Medicinal plants are used for the ailment of several microbial and
non-microbial originated diseases due to their valuable effects in health care
(Akroum et al., 2009). The
affordability, reliability, availability and low toxicity of medicinal plants
in therapeutic use has made them popular and acceptable by all religions for
implementation in medical health care all over the world. Plants are indeed the
first material used in alternative medicine type of remedy against many
diseases (Amjad et al., 2005). Herbal
medicine has been widely used as an integral part of primary health care in
many countries (Akinyemi et al.,
2005). Medicinal plants may constitute a reservoir of new antimicrobial
substances to be discovered. The World Health Organization (WHO) reported that
4 billion people (80% of the world’s population) use herbal medicines for some
aspect of primary healthcare (Fabricant and Farnsworth, 2001). In developing
countries, 80% of people use traditional medicines which are based on plant
products. Thus many studies have been conducted on medicinal plants. Currently
80% of the world population depends on plant derived medicine for the first
line of primary health care for human alleviation because it has no side
effects (Chaudhari et al., 2012).
Plants are important source of medicines and presently about 25% of
pharmaceutical prescriptions in the United States contain at least one plant derived
ingredient (Pandey et al., 2011). In
the last century, roughly 121 pharmaceutical products were formulated based on
the traditional knowledge obtained from various sources (Hasan et al., 2009).
Bacterial
infections are one of the prominent causes of health problems, physical
disabilities and mortalities around the world. Medicinal plants are a rich
source of antimicrobials and provide a safer and cost effective way of treating
bacterial infections. The antibacterial activity of natural products from medicinal
plants is applicable for the treatment of bacterial, fungal and viral diseases.
Medicinal plants have been recognized as potential drug candidates because they
possess drug like properties. Even though pharmacological industries have
produced a number of new antibiotics in the last three decades, resistance to
these drugs by microorganisms has increased (Bernhoft, 2010). Secondary
metabolites or phytochemicals such as phenols, flavonoids, alkaloids,
terpenoids, and essential oil have proved to be responsible for the
antimicrobial activity of plants (Hwang et
al., 2001). Phenol and polyphenol group of compounds consist of thousands
of diverse molecules with heterogeneous structure with common feature of having
one or more phenol ring. Several workers have reported that phenolic compounds
such as gallic acid, coumarins, polyphenols, caffeic acid, cinnamic acid,
pyrogallol, eugenol show antimicrobial activity against virus, bacteria and
fungi (Saify et al., 2005).
Of
all the plant spread by nature upon the surface of the globe, there are none
more beautiful than those we know under the name of citron, lemon, and orange
trees which botanists have included under the technical and generic name
Citrus. No other equals them in beauty of leaf, delightful odor of flower, or
splendor and taste of fruit. These brilliant qualities have made the Citrus a
favorite in all countries. The genus Citrus, belonging to the Rutaceae family,
comprise of about 140 genera and 1,300 species. Citrus sinensis (Orange),
Citrus paradise (Grapefruit), Citrus limon (Lemon), Citrus reticulata
(Tangerine), Citrus grandis (Shaddock), Citrus aurantium (Sour orange), Citrus
medica (Citron), and Citrus aurantifolia (Lime) are some important fruit of
genus Citrus (Anwar et al., 2008).
Citrus are well known as one of the world’s major fruit crops that are produced
in many countries with tropical or subtropical climate. Brazil, Japan, China,
Mexico, Pakistan, and countries of the Mediterranean region, are the major
Citrus producers. The Citrus fruits and their by–products are of high economic
and medicinal value because of their multiple uses, such as in food industry,
cosmetics and folk medicine (Silalahi, 2002). In additions to large scale
consumption as fresh fruits, the fruits are mainly processed to produce juice.
The waste of Citrus processing industry left after juice extraction, such as
peels, seeds and pulps, corresponding to about 50% of the raw processed fruit,
can be used as a potential source of valuable by products. Specifically, the
Citrus peels, commonly treated as agro-industrial waste, are a potential source
of valuable secondary plant metabolites and essential oils (Andrea et al., 2003).
1.1 AIM AND OBJECTIVES
The
aim of this study was to determine the antibacterial effects of sweet orange (Citrus sinensis) and lime (Citrus aurantifilia) against Pseudomonas aureginosa, Staphylococcus aureus, Escherichia coli
and Salmonella typhi by agar well
diffusion method.
The objectives
are;
1. To
determine the antibacterial effects of sweet orange (Citrus sinensis) and lime (Citrus
aurantifilia) against some selected pathogens
2. To
determine the MIC of sweet orange (Citrus
sinensis) and lime (Citrus
aurantifilia) extracts
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