ABSTRACT
This study was to access the antimicrobial and antioxidant properties of Tetraplera tetraptera on some human pathogens. The antimicrobial activity of Tetraplera tetraptera on some pathogens was investigated with different solvents (Ethanol, Methanol, Cold and Hot water) against test organisms (Esherichia coli, Staphylococcus aureuas, Salmonella spp, Shigella, Aspergillus and Candida albican). The different extracts were prepared at different concentration (50mg/ml, 100 mg/ml, 150mg/ml and 200mg/ml). Amongst the different Tetraplera tetraptera extracts studies, ethanolic extract generated more phytochemical constituents than the methanol, cold water and hot water extracts. The study revealed that the ethanolic extract had more significant inhibitory effect than methanolic, cold water and hot water extracts. Salmonella spp. and Staphylococcus aureus, had the highest sensitivity (15mm, and 16mm) against the methanol and ethanol extract of Tetraplera tetraptera, followed by Aspergillus niger on the ethanolic extracts of Tetraplera tetraptera at 14mm. Ciprofloxacin and Chloramphenicol were used as control. The antioxidant potentials of Tetraplera tetraptera plant extracts have been linked with their flavonoids contents may provide anti-inflammatory activity. Ethanolic extracts and methanolic extracts of Tetraplera tetraptera have broad-spectrum antimicrobial activity; hence can serve as natural therapeutic agent against some enteric pathogens.
TABLE OF CONTENTS
Title Page i
Certification iii
Dedication iv
Acknowledgement v
Table of Contents vi
List of Tables vii
Abstract ix
CHAPTER ONE
Introduction 1
1.1 Aim and Objectives 4
CHAPTER TWO
Literature
Review 6
2.1 Description of the Plant and Use 6
2.2 Classification of Phytochemicals 7
2.2.1 Tannins 7
2.2.2 Flavonoids 7
2.2.3 Organic Acids 8
2.2.4 Phenols 8
2.2.5 Nitrogenous Compounds 8
2.2.6 Terpenes or Terpenoids 8
2.2.7 Alkaloids 9
2.2.8 Saponins 9
2.3 Antioxidant
Properties of Phenolic Compounds 10
2.4 Antioxidant
Activity of Aromatic Plants 12
2.5 Antioxidant
Activity from Aroma Compounds and Essential Oils 13
2.6 Essential
Oils as Antioxidants in Foods 16
2.8 Biology and Pathogenecity of Test
Organisms 18
2.8.1 Escherichia
coli 18
2.8.2 Salmonella 18
2.8.3 Shigella 19
2.8.4 Candida
albicans 19
2.9 Comparative Study of the Phytochemical
Composition of the Leaves of
Five Nigerian Medicinal Plants 20
CHAPTER THREE
Materials and Methods 21
3.1 Sample Collection 21
3.2 Sterilization of Materials 21
3.3 Preparation of Extracts 21
3.3.1 Ethanolic and Methanolic Extract Preparation 21
3.3.2 Preparation of Different Concentration of
Extract 22
3.4 Biochemical Identification of the Test
Organisms 22
3.4.1 Catalase Test 22
3.4.2 Indole Test 22
3.4.3 Citrate Utilization Test 22
3.4.5 Motility Test 23
3.4.6 Voges-Proskauer Test 23
3.4.7 Methyl Red Test 23
3.5 Determination of Antimicrobial Activity 24
3.6 Determination of Minimum Inhibitory
Concentration (MIC) 24
3.7 Determination of Minimum Bactericidal Concentration 25
3.8 Phytochemical Screening of the Extract 25
3.8.1 Qualitative
Analysis 25
3.9 Methods
for Testing Antioxidative Activity 26
3.10 Determination of Minerals 26
3.10.1 Calcium and Magnesium Determination 26
3.11 Determination of Vitamins 27
3.11.1 Thiamine (VIT B1) Determination 27
3.11.2 Riboflavine (VIT B2) Determination 28
3.11.3 Niacin Content Determination 28
3.11.4 Determination of Vitamin C 29
CHAPTER FOUR
Results 28
4.1. Qualitative Phyto-Chemical Constituents of
Tetrapleura tetraptera Leave Extract 30
4.2. Identification and Characterization of
Indicator Bacterial 30
4.3. Identification
and Characterize of Test Organisms (Fungal) 30
4.4. Antimicrobial Activity of Tetrapleura Tetraptera Plant Extract against
Fungi,
Gram Positive and Negative Human
Pathogen 30
4.5a. Minimum Inhibitory Concentration and Maximum
Bactericidal Concentration
Value of Tetrapleura Tetraptera against Test Bacteria and Fungi 31
4.5b. Minimum Bactericidal Concentration Value of Tetrapleura Tetraptera against
Test Bacteria and Fungi 31
4.6. Vitamin Composition
Analysis of Tetrapleura Tetraptera
Plant Leaf 32
CHAPTER FIVE
5.0 Discussion, Conclusion and Recommendation 40
5.1 Discussion 40
5.2 Conclusion 42
5.3 Recommendation 42
References
LIST OF TABLES
|
TABLE
|
TITLE
|
PAGE
|
|
1
|
Qualitative Phyto-Chemical Constituents of Tetrapleura tetraptera Leave Extract
|
33
|
|
2
|
Identification
and Characterization of Indicator Bacterial
|
34
|
|
3
|
Identification and Characterize of
Test Organisms (Fungal)
|
35
|
|
4
|
Antimicrobial Activity
of Tetrapleura Tetraptera Plant
Extract Against Fungi, Gram Positive and Negative Human Pathogen
|
36
|
|
5a
|
Minimum Inhibitory
Concentration Value of Tetrapleura
Tetraptera against Test Bacteria and Fungi
|
37
|
|
5b
|
Minimum Bactericidal
Concentration Value of Tetrapleura
Tetraptera against Test Bacteria and Fungi
|
38
|
|
6
|
Vitamin
Composition Analysis of Tetrapleura
Tetraptera Plant Leaf
|
39
|
CHAPTER ONE
1.0 INTRODUCTION
The
use of these plant materials and as preservative and as a means of preventing
microorganism development in foods has become the subject of extensive studies
(Gould, 2006).Importantly, the inhibitory effects of Tetrapleura tetraptera
extracts against some human pathogens have been reported. Currently, there is a
growing demand worldwide of consumers for minimizing chemical preservation that
can be detrimental to human health consequently, spices, herbs and naturally
occurring phenolics from various plants sources are being studied in detail in
response to consumer requirements for fresher and more natural additive-free products
(Nychas, 2005)
Tetrapleura
tetraptera is one of the medicinal plants in Nigeria.
The documented biological or pharmacological activities are found to be
molluscicidal, cardiovascular, neuromuscular, hypotensive, anti-conversant,
anti ulcerative, anti-inflammatory and anti-microbial. The pods notably have an
appealing culinary use for mothers from the first day of delivery to post
parturition and as a lactation aid (Enwere, 2008).The antibacteial activity of
this plant has been formulated into soap bases using palm kernel oil. At the
same time, most of the folkloric chains agree in the traditional use of the
fruit for management of convulsion, leprosy, inflammation and rheumatoid pains
(Dalziel, 2008).
Alcoholic
and water extracts of Tetrapleura tetraptera inhibited the growth of Staphylococcus
aureus (Salako et al., 2010).The antibacterial activities of
these plants has been exploited in the formulation of the dried powdered
fruit of the plant, thus, dried powdered herbs have been formulated
into soap bases using palm kernel oil, shea butter and mixture of bases.
The formulated soaps were evaluated for organoleptic properties and
foaming ability. Soaps with mixture of these two bases were of better
qualities than those with the individual base. Incorporation of powdered
plant materials influenced both the foaming property and the hardness
of the soaps. Except for the Tetrapleura tetraptera fruit powder which
improved the foaming ability of these soaps all other herbs
including Acalypha wilkensia,Harugana madagascariensis and Ficus
exasperate depressed the foaming ability of the soaps. The extract
from Tetrapleura tetraptera exhibited anti-convulsant activity, which
could be linked to their ability to depress the central nervous
system (Akah and Nwambie, 2003). The ethanol extract and saponins from
stem-bark of Tetrapleura tetraptera exerted an inhibitory effect on
luteinizing hormone released by pituitary cells (El Izziet et al.,2010)
suggesting its use as a contraceptive agent. The nutritive quality of
the dry fruit of Tetraptera tetraptera used as spice was
assessed. The fruit shell, fruit pulp and seed contained varying amount
of nutrients such as proteins, lipids and minerals which were comparable
and some were higher than popular spices such as red pepper, onion curry
and ginger (Essien et al., 2004)
Reactive oxygen species and free radicals formed during
oxidation have been reported to contribute to diseases such as cancer,
diabetes, cardiovascular diseases and ageing (Halliwell and
Gutteridge, 2009). Anitoxidants have the ability to
protect the body from oxidative damage (Ozsoy et al., 2008) by scavenging the free
radicals and inhibiting peroxidation and other radical mediated processes. In
recent years, significant attention has been directed towards exploring
plant-based natural antioxidants, especially the phenolics and tocopherols (Chaovanalikit
and Wrolstad, 2004). Such natural antioxidants are
not only reported to have anti-carcinogenic potential that protects the foods
from oxidative deterioration but also, these are associated with other health
beneficial effects such as, lowering the incidence of aging, inflammation,
cardiovascular diseases and certain cancers (Iqbal et al., 2007). Various antioxidant
activity methods have been used to monitor and compare the antioxidant activity
of food (Halliwell and Gutteridge, 2007).
They may be free radicals which possess an unpaired electron in their outermost
shell and are capable of independent existence (Halliwell and
Gutteridge, 2007). Their half-lives vary from a few
nanoseconds for the most reactive compounds to seconds and hours for rather stable
radicals. They trigger chain reactions resulting in the oxidation of
macromolecules in order to reach a steady state (Aristidis et al., 2012). Polyphenolic compounds constitute a crucial category of
antioxidant metabolites. They are plant secondary metabolites which have at
least one aromatic ring in their molecule and usually exist in the form of
glycosides. They protect plants against harmful environmental conditions and
the attack of microorganisms and contribute to the development of several characteristics
such as color (Manach et al., 2004). Tetrapleura tetraptera and
Parkia biglobosa are two
widely utilized plants in West Africa for their perceived nutritional and medicinal value (Adetunji and Aladesanmi, 2006).The fruits of both plants are
used in foods for flavor and seasoning of different traditional
dishes. Ethnomedical practices throughout West Africa use these plants for the
treatment of several ailments including arthritis, asthma, diabetes mellitus, hypertension, epilepsy,
schistomiasis, and even prevention of post-partum contraction (Ojewole et al., 2004) and gastro-intestinal
disorders are attributable to the bioactivity of phytochemicals in the fruits of Tetrapleura
tetraptera (Adetunji and Aladesanmi, 2006). The
phyto constituents
of the fruits of Parkia biglobosa are
also used in West African ethnomedicine to treat leprosy and
hypertension. Taken together, anecdotal evidence from nutritional supplementation and
ethnomedicinal practices suggests that the biologically active molecules in the fruits of Tetrapleura
tetraptera and Parkia biglobosa are present
a potential source of drug leads.
Prior studies have confirmed the presence of glycosides,
flavonoids, saponins, essential oils, saponosides, triterpenes, coumarins,
tannins, sugars, steroids, tritepene glycosides, tannins, and polyphenolic compounds
as phyto constituents in the fruits of Tetrapleura tetraptera and Parkia
biglobosa. The fruit of Tetrapleura tetraptera shows an additional
presence of alkaloids (Ojewole et al., 2004). Since tannins, flavonoids
and polyphenols represent molecules generally known for antioxidant
bioactivities
The fruit of Tetrapleura tetraptera is used by herbalist to treat various
ailments such as diabetes mellitus, hypertension, epiley, schistosmiases, etc (John et al.,
2004). It is used extensively in soup giving to
nursing monthers to prevent post partum contractions (Nwawu and Alah,
2006) and gastrointestinal disorders, including
stomach ulceration (Noamesi et al,
1992).The fruit is said to contain caffeic acid which is reported to have anti-tumour
activity, anti-inflammatory properties, and also inhibit
HIV replication. It is therefore necessary to evaluate
phytochemical composition, antioxidant and antimicrobial properties of the Tetrapleura tetraptera, to diversity
their use as natural preservatives and as culinary spices that contain active
ingredients that promote health and reduce the risk of disease.
Preliminary studies indicate that the
incorporation of Tetrapleura tetraptera
fruit in the diet for both men and women, and as well as during pre and post-delivery
could be beneficial for relieving of many ailments. The paucity of knowledge of the phytochemical
constituents, antioxidant
and antimicrobial properties of Tetrapleura
tetraptera has resulted in their neglect and under utilization. There is therefore, the need to study further, the effects of the fruits of TTE on microorganisms
1.1 AIM
AND OBJECTIVES
To
Assess the Antimicrobial and Antioxidant of Tetrapleura
Tetraptera on Some Human Pathogens, while the specific objectives are;
· To
determine the antibacterial effects of Tetrapleura tetraptera extract on
selected isolates.
· To
characterized the phytochemical composition of the ethanol extracts of Tetrapleura
tetraptera
· To
determine the antioxidant properties of Tetrapleura tetraptera extract
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