ABSTRACT
The upsurge in the prevalence of side effects of many synthetic antimicrobial agents and incidence of multidrug resistant bacteria has spurred scientists on the research for plant based antimicrobial of therapeutic potentials. Azadirachta indica and Psidium guajava present such potential of high medicinal value. These plants are used in Nigeria traditionally as condiments and for treatment of various ailments such dental plague . The phytochemical composition of Azadirachta indica and Psidium guajava showed the presence of Tannin, Alkaloid, Saponin and Flavonoid. Aqueous and ethanol extracts of Azadirachta indica and Psidium guajava leaves were screened for antibacterial activity against Candida albican and Streptococcus mutans. Both extracts were found to exhibit selective inhibition against the isolates. The diameter zones of inhibition exhibited by the extracts were between 2 ± 0.01 – 10 ± 0.10 mm. The Minimum Inhibitory Concentration (MIC) determined by the agar diffusion method was between 10.00 and 2.50 mg/ml-1. Ethanol extracts showed more `inhibitory effect compared to the aqueous extracts. Results obtained show that the extracts of Azadirachta indica and Psidium guajava possess some level of antibacterial activities against Steptococcus mutans and Candida albicans.
TABLE OF CONTENTS
Title
Page i
Certification
ii
Dedication
iii
Acknowledgements
iv
Table
of Contents v
Lists
of Tables vii
Abstract
viii
CHAPTER ONE
1.0 Introduction 1
1.1
Aims and Objectives 4
1.2 Objectives
4
CHAPTER
TWO
2.0 Literature
Review 5
2.1 Dogoyaro/Neem
(Azadirachta indica) 5
2.2 Phytochemicals
of Azadirachta
indica 8
2.3 Antimicrobial
Activity 9
2.4 Antibacterial
and Anti-Fungal Activities 10
2.5 Guava
(Psidium guajavaLinn.) 10
2.6
Streptococcus
mutans 14
2.7
Candida
albicans 15
CHAPTER THREE
3.0 Materials
and Methods 17
3.1 Collection of Samples 17
3.2
Media
Used 17
3.3 Sterilization 17
3.4 Collection of Test Organisms 17
3.5 Isolation and Identification of Bacteria and Fungi 17
3.6 Preparations
of Plant Extracts 18
3.7 Phytochemical Analysis 18
3.7.1 Test for alkaloid 18
3.7.2 Test for flavonoid 19
3.7.3 Determination of saponins 19
3.7.4 Test for tannin 20
3.8 Antibiotic Sensitivity Testing 20
3.8.1 Agar diffusion method 20
3.9 Determination of Minimum Inhibitory
Concentration (MIC) 20
CHAPTER FOUR
4.0
Results 22
CHAPTER FIVE
5.0 Discussion Conclusion Recommendation 27
5.1 Discussion 27
5.2 Conclusion 28
5.3 Recommendation 28
References 29
LIST OF TABLES
Table
Title Page
1:
Antibacterial activity
of the aqueous of Azadirachta
Indica and Psidium guajava against Streptococcus
mutans and Candida albicans 23
2:
Antibacterial activity
of the ethanol extract of Azadirachta
Indica and Psidium guajava against Streptococcus mutans
and Candida albicans 24
3: Phytochemical composition of Azadirachta
Indica and Psidium guajava 25
4: Minimum inhibitory concentration
(MIC) of the aqueous and
ethanol
extracts of Azadirachta indica and Psidium guajava 26
CHAPTER ONE
1.0 INTRODUCTION
The microorganisms present in the mouth
consist of a diverse and populous collection of bacteria, fungi, and transient
viruses. Bacteria make up the largest number of varieties (Akinyele et al.,2014).
More than 350 cultivable bacterial species have been identified in the oral
cavity and molecular analyses suggest that an equal number of non-cultivable
flora are also present (Marcotte and Lavoie, 1998). Bacterial accumulation on oral
surfaces is a major factor in the development of most of the common dental
diseases such as dental caries, plaque and periodontal disease (Vohra et
al.,2012).
Dental plaque is a complex biofilm found
on the tooth surface which forms the major reason of the development of dental
caries. The accumulation and development of plaque depends upon the outcome of
the interactions between the adhesiveness of plaque to the tooth surface and
the physical shear forces which serve to dislodge and remove the plaque (Vohraet
al.,2012). Tooth brushing with toothpaste is the most widely practiced form
of oral hygiene in most countries (Pannutiet al.,2003) and the success
of any toothpaste, in part, lies on its ability to eliminate pathogenic oral
microflora. A wide range of chemicals, mainly antimicrobial agents, have been
added to toothpastes in order to produce a direct inhibitory effect on plaque
formation (Fine etal., 2006). Fluoridated toothpastes have been proven
to protect teeth against attack from bacteria (Vohraet al.,2012).
Nigerians basically employ two
methods to remove debris (plaque) from the mouth which are either by use of
tooth brush and paste or by use of parts of various plants native to West
Africa, referred to as “African chewing sticks” (Bankoleet al.,2012).
Meanwhile, in absence of toothpaste, a large proportion of persons living in
rural areas and some who are also in the urban areas make use of the natural
toothbrush known as “chewing sticks. Ojoet al.,(2007) defined chewing
sticks as parts of higher plants which are cut into suitable lengths and used
for the maintenance of oral hygiene. They are important Non Timber Forest
Product (NTFP) widely used for dental cleaning in the tropical West Africa
(Akande and Hayashi, 1998). Plants from which chewing sticks are derived are
abundant and diverse in Nigeria rural communities. Almost the entire rural
population in Nigeria use chewing sticks for orodental hygiene (Bankoleet
al.,2012).
This is due to the fact that the
Nigerian chewing sticks, are readily available, cheap and efficacious (Kareem et
al.,2012). Chewing sticks are recommended for oral hygiene by the World
Health Organization, and some of them, or their extracts, are also used in
ethno-medical treatment of oral infections (Ndukwe, et al.,2005).
Several studies have demonstrated the antiplaque and antibacterial activities
of extracts of these Nigerian Chewing Sticks (NCS) against oral bacteria, such
as Streptococcus mutans and Streptococcus mitiswhich are
organisms commonly implicated in dental caries and orodental infections. The
plants used as chewing sticks are carefully selected for certain properties
such as foaminess, hardness, or bitterness and certain species are more popular
than the others, in which a great number of these plant species have related
medicinal properties that may be antibacterial and are as important today as
they were thousands of years ago (Hullins, 2010).
In addition, Hoodaet al.,(2010)
reported that extracts of chewing sticks can be incorporated into tooth pastes or
used as mouth washes. The choice of chewing sticks to be used in most cases
depends on its cleansing action of the teeth; the therapeutic value, or
preferred taste or flavour. The sticks (which may be stem or root with bark
removed or retained) are cut to convenient lengths and washed thoroughly with
fresh water to get rid of any dirt (Ogundiyaet al.,2006). The usefulness
of chewing sticks in oral hygiene maintenance has been considered comparatively
effective as tooth brush (Van-Palentsteinet al.,2002).
Oral diseases impact our
quality of life and may lead to systemic and threatening diseases. The
relationship between the high incidence of oral diseases and microorganisms is
well known. Because of the increased microbial resistance to antibiotics, toxic
and harmful effects of few common antimicrobial agents, there is a continuous
need for alternative therapies which areaffordable, non toxic and effective,
such plants (Van-Palentsteinet al.,2002).
Alternative medicines are
commonly included in therapeutic and diagnostic disciplines outside the
conventional health system(Bankoleet al.,2012). In developing countries, access to dental healthcare is
restricted and expensive and thus it is limited to emergency dental care
procedures (Bankoleet al.,2012).
Dental caries is o most
common infection of all oral diseases. It is proved that cariogenic
microorganisms, especially Streptococcus mutans plays an essential role
in the pathogenesis of dental caries. It is involved in the initiation of
almost all carious lesions in enamel (Bankoleet
al.,2012).
The main cause of dental
caries is attributing to oral biofilm, also known as dental plaque, a film of
microorganisms sticking to the tooth surface
(Bankole et al.,2012). Streptococcus mutanshas
the ability to metabolize dietary sucrose and synthesize glucan by cell surface
and extracellular glucosyl transferase. This glucan is an insoluble sticky or
slimy gel relatively inert and resistant to bacterial hydrolytic enzymes which
causes plaque to adhere tenaciously to tooth surface (Akinyeleet al.,2014). Streptococcus mutans organisms in the plaque
produce organic acids such as lactic acid that gradually destroy the enamel to
form a cavity to Streptococcus mutans, Lactobacillus acidophilus
probably also play role in acid production in the plaque and involved in root
caries and periodontal diseases Enterococcus faecalis (Akinyeleet
al.,2014).
Candida
albicans, are some other microbial
species that knowingly cause several oral diseases, such as dental caries,
endodontic infections, periodontal diseases and oral Candidiasis. The most effective method to eradicate C.albicans is
the use of sodium hypochlorite and 2% chlorhexidine(Van-Palentstein
et al.,2002).
1.1
AIMS AND OBJECTIVES
The aim of this research is to study
the antimicrobial effect of Azadirachta indica (Dogoyaro /Neem) and Psidium guajava L. (Guava) on Streptococcus
mutans and Candida albican.
1.2 OBJECTIVES
1.
To determine the phytochemical
constituents of Azadirachta indica
(Dogoyaro/Neem) and Psidium guajava L. (Guava) .
2. To determine the antimicrobial activity of Azadirachta
indica
(Dogoyaro/Neem) and Psidium guajava L. (Guava)on Streptococcus
mutans and Candida albican.
3. To determine the minimal inhibitory concentration
of Azadirachta indica (Dogoyaro/Neem) and Psidium guajava L. (Guava)on Streptococcus
mutans and Candida albican.
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