ABSTRACT
The antimicrobial activities of the fruit parts extracts of Chrysophyllum canito and micro organisms associated with its spoilage. Bioactive compounds were extracted from the fruit parts using different solvents (Aqueous, Ethanol and Ethylacetate. Preliminary phytochemical analysis reveals the presence of alkaloid, Saponin, Tannin, Cyanogenic glycoside, Flavonoid, Steriod and starch. Antioxidant activities obtained were scavenging at concentration of 25% of the seed, chelating at 25% of the seed and reducing at 43% of the seed while the pulp possessed scavenging at concentration of 50.90%, chelating at 85.63% and reducing at 39.23%. the peel on the other hand possessed scavenging at concentration of 34%, chelating at 41% and 27% of reducing, therefore it is been deduced that the pulp contained more of the chelating antioxidant property. Using disc diffusion method each prepared extract was tested on the different isolated organisms and the extent of sensitivity of the tested organism and the extent of sensitivity of the tested organism on the different extracts was measured by their zones of inhibition.
Aqueous extract of the peel inhibited growth of the test organism at the highest range of 11.67 to 21.67mm, Ethanol at the range of 16.00 to 20.00mm, Ethyl acetate at the range of 6.67 to 12.33. the pulp recorded its water extract to inhibit growth of the test organism at the highest range of 10.33 to 17.33mm, Ethanol at the range of 12.67 to 15.33mm E acetate at the range of 6.67 to 7.67 while the water extract of the seed inhibited growth of the test organism at the highest range of 14.00 to 23.67mm, Ethanol at the range of 17.33 to 20.00mm, Ethyl acetate at the range of 9.33 to 12.33. a standard broad spectrum antibiotics (ciprofloxacin) was used as a control. The action of different extracts shows variation of significant difference (P< 0.05). The lowest minimum inhibitory concentration of the extracts was at the concentration 100mg/mol and the highest minimum inhibitory concentration was at 200mg/mol. Organisms obtained from the culture of the spoilt udara were Staphylococcus aureus, Bacillus, Pseudomonas aerogenosa, Lactobacillu and Proteus and fungi Aspergilus, Penicillium, Fusarium, Rhizopus, Trichoderma and Bortyodiplodia. There is basis for the traditional use of extracts of Chrysophyllum canito fruits as a local health remedy for system infections and the result of this study can be a precursor for the production of drugs for the body system infections.
TABLE OF CONTENTS
i
Certification ii
Dedication iii
Acknowledgments iv
Table of
contents v
List of
Tables x
List of
Figures xi
Abstract xii
CHAPTER ONE
1.0 Introduction 1
1.1 Aims and Objectives 4
CHAPTER TWO
2.0 Literature Review 5
2.1 Udara fruit 5
2.2 Native name of udara 6
2.3 Health benefits of
African star apple 6
2.3.1 Vitamins and minerals 7
2.3.2 Helps Diabetic condition 7
2.3.3 Medicinal uses 7
2.3.4 Help you lose weight 8
2.3.5 All round basic nutrition 8
2.4 Spoilage 8
2.5 Fruit spoilage prevention 9
2.5.1 Mechanical damage 9
2.6 Natural decay 9
2.7 Micro organism
contamination 10
2.8 Fruit spoilage
intervention 10
2.19 Botanical fruit and culinary
fruit 11
2.10 Fruit Structure 11
2.11 Fruit Development 11
2.12 Phytochemicals of Chrysophyllum cainito 13
2.12.1 Saponins 13
2.12.1.1 Health benefits of Saponins 14
2.12.2 Alkaloids 15
2.12.2.1 Functions of alkaloids 15
2.12.3 Tannins 16
2.12.3.1 Benefits of Tannins 16
2.12.4 Flavonoids 16
2.12.4.1 Health benefits of flavonoids 17
2.12.5 Glycosides 17
2.12.5.1 Health benefits of glycosides 18
2.13 Antioxidants 18
2.13.1 DPPH (2,2-Diphenyl -1-Picryl –
hydrazyl – hydrate) 19
2.13.2 FRAP (Ferric Reducing
antioxidant power) 19
2.14 Antimicrobial 20
2.15 Factors to consider while
choosing an antimicrobial agent. 21
CHAPTER
THREE
3.0 Materials and Methods 22
3.1 Materials 22
3.2 Methods 22
3.2.1 Sources of samples 22
3.3 Sampling and sample
preparation 22
3.4 Collection of test
organism 23
3.5 Sterilization of
equipment/materials 23
3.6 Preparation of sample for
extraction 23
3.7 Production of extracts 24
3.8 Phytochemical test 24
3.9 Phytochemical spot test
(Qualitative analysis) 24
3.9.1 Qualitative analysis of
phytochemicals 25
3.10 Quantitative determination
of phytochemicals 28
3.11 Antioxidant determination 31
3.11.1 2,2, Diphenyl – 1 – picry
Hydrazyl (DPPH) Phytometric assay 31
3.11.2 Ferric reducing antioxidant
power (FRAP) 32
3.11.3 Ferrous ion-chelating ability 32
3.12 Media preparation 32
3.13 Preparation of different
concentration of the fruit extract and disc impregnation 33
3.14 Antimicrobial assay 33
3.15 Determination of MIC 34
3.16 Preparation of sample for
microbial isolation 34
3.17 Pathogenicity test 35
3.18 Isolation of microbe from
spoilt udara fruits 35
3.19 Spread plate culture 35
3.20 Serial Dillution 36
3.21 Direct microbial culture
(For fungi) 36
3.22 Purification of isolate (sub
cultures) 37
3.23 Characterisation of isolates 37
3.23.1 Characterisation of fungal
isolates 37
3.23.2 Characterisation of bacterial
isolates 38
3.23.2.1 Gram stain reaction 38
3.23.2.2 Carbohydrate utilization test 39
3.24 Identification of Bacteria
and fungi 41
3.25 Statistical Analysis 42
CHAPTER FOUR
4.0 Results 43
CHAPTER FIVE
5.0 Discussion 57
5.1 Conclusion 60
5.2 Recommendation 61
Reference
Appendixes
LIST OF TABLES
TABLES TITLE PAGE
1 Taxonomical classification of Chrysophyllum cainito 6
2 Qualitative phytochemical screening
of parts of udara fruits 46
3 Antioxidant table activity 47
4 antimicrobial activities of Chrysophyllum cainito 48
5 Incidence of occurrence of fungal
isolates on C. cainito stored in nine
days 50
6 Occurrence of bacterial isolates
from spoilt udara fruit 52
7 Occurrence of fungal isolates from
spoilt udara fruit 53
8 Characteristics of bacterial
isolates from spoilt udara fruit 54
9 Characteristics of fungal isolates
from spoilt udara fruit 55
LIST OF FIGURES
FIGURES TITLE PAGE
1 Minimum
inhibitory concentration of C. cainito extract 49
2 Incidence
of occurrence and pathogenecity of fungi 51
CHAPTER ONE
1.0
INTRODUCTION
Worldwide,
infectious diseases are the leading causes of death especially in tropical
countries. The resistance of micro organisms to antibiotics coupled with the
emergence of unknown disease causing micro organisms have inspired scientists
to search for new antimicrobial agents.
Since time
immemorial, man has sufficiently and efficiently use parts of fruits to treat
and prevent diseases. Researchers are increasing their attention to curative
medicine.
In common language usage, “fruit” normally
means the fleshy seed associated structures of a plant that are sweet or sour,
and edible in the raw state, such as apples (star apple), bananas, grapes,
lemons, oranges and strawberries. Chrysophyllum canito (inn) belongs to the family sapotaceae. It
is primarily a forest tree species and its natural occurrences have been
reported in diverse ecozones in Nigeria, Uganda, Niger Republic, Cameroun and Cote
d’Ivoire (Bada, 1997). The plant often grows to a height of 36.5m though it may
be smaller (Bada, 1997). The African start apple fruit is a large berry
containing 4 to 5 flattened seeds or some times fewer due to seed abortion
(Adisa, 2000). The plant has in recent times become a crop of commercial value
in Nigeria. The fleshy pulp of the fruits is eaten especially as snack and
relished by both young and old (Cenrad, 1999). The African star apple fruit has
been found to have highest content of ascorbic acid with 1000 to 3,330mg of
ascorbic acid per 100gm of edible fruit or about 100 times that of oranges and
10 times of that of guava or cashew (Asenjo, 1946). It is reported as an
excellent source of vitamin, irons, flavours to diets and raw materials to some
manufacturing industries (Adisa, 2000; Bada, 1997). In addition, its seeds are
a source of oil, which is is used for diverse purposes. The seeds are also used
for local games (Bada, 1997). The fruits also contain 90% anacardic acid, which
is used industrially in protecting wood and as source of resin, while several
other components of the tree including the roots and leaves are used for
medicinal purposes (Adewuesi, 1997)
The fruit is common in both urban and
rural center especially during the months of December to April. The fruits are
not usually harvested from the trees, but left to drop naturally to the forest
floor floor where they are picked. Allowing the fruits to drop before picking
promotes fungal infections. Recent market survey revealed that the fruits often
deteriorate within a very short period. According to National Research Council,
2008 Chrysophyllum albidum actually
becomes bad in a period of 5 days, with the deterioration starting with the
change of color from uniform orange to one with patches and followed by
shrinking of the fruit. This study was undertaken to investigate the etiology
of post-harvest biodeterioration of African star apple fruits in Umudike, Umuahia,
Abia State, Nigeria and the effects on its nutrient (food) value. It is
commonly known as “Agbalumo” or “Osan” (Yoruba) or “Udala” (Igbo) in the local
languages. Its fruit which is pale yellow with pink coloured endocarp is
relished by both children and adults when in season. Its fully ripe fruit
becomes available from January through March in southwestern parts of Nigeria.
The pink coloured pulp and the whitish cover of the brown-coloured seeds of the
fruit are consumed while the empty pale yellow pericarp is discarded.
It is sparsely
distributed in Nigeria; grows rapidly and reaches 20m in height (Luo et al, 2002). It has numerous common
names including cainito, caimils star apple, golden leaf, tree and also milk
fruit (National Research Council, 2008). The fruit has antioxidant properties
(Luo et al, 2002; Einbond et al., 2004). It is of great important
economic value due to its diverse industrial, medicinal and food uses. The
fruits are not only consumed fresh but also used to produce jam, jellies,
stewed fruit, marmalade, syrup and several types of soft drinks. It is also
used for medical purposes due to properties of stalk and fruits. The leaves and
seed of some of these fruits and vegetables are used in pharmaceuticals. Some
of the trees are also valuable for ornamentation as an evergreen bread leaf
plant (Islam 2002). The bark is considered a tonic and stimulant and a bark
decoction is used as an antitussive (Luo et
al., 2002).
Home gardens combine physical,
social and economic functions on the land around the family home in a form of
traditional agro forestry. Home gardens supplement food and fruit supplies from
farms that are far from the settlement (Aweto, 2014). In tropics, the culture
of keeping home garden edible fruits, commonly used medicinal plants for
pediatrics ailments and other food items is a common practice especially. In
Africa, Asia and Brazil. Edible fruit trees (EFTs) are equally important in the
environmental amelioration and carbon sequestration of urban centres in
developing countries. Edible fruit tree species are either cultivated or semi
wild and protected around homesteads in rural, peri-urban and urban communities
in Nigeria. They improve the aesthetic value of the environment, render
ecological and social services, play indirect roles in improving mental,
psychological health and social well-being. These edible fruit trees and other
plants contribute to the urban greening and plant diversity in a large extent
besides their food functions.
The Ajurvedic system of medicine has
described various fruits in the treatment of diseases which play an important
role in modern health care and curing various several reports on the chemical
composition and antimicrobial activities of some fruits and their extracts that
inhibits various bacteria (Luo et al.,
2002). However studies on the chemical composition and antimicrobial activities
of Chrysophyllum cainito are limited.
Therefore, scientific evaluation of this fruit is important to elucidate its
antimicrobial effect and micro organism associated with its spoilage.
1.1 AIMS
AND OBJECTIVES
Fruits can be used
as drugs, food and flavors. This research work is aimed at the evaluation of
the antimicrobial effects of the seed, peel and pulp of Chrysophyllum cainito and some microbes associated to its spoilage
in which includes : bacteria isolates which are species of Bacillus, staphylococcus, lactobacillus, proteus and pseudomonas. The fungal isolates
includes species of Rhizopus, Aspergillus, Fusarium, Trichoderma, Botryodiplodia and Penicillium ..
·
To
elucidate its antimicrobial effects in alternative medicine in the treatment of
some infections especially enteric diseases.
·
To
determine the antimicrobial effect of Chrysophyllum
canito fruit and the
microbes associated to us spoilage.
·
To
isolate, purity and characterize microbial isolated from spoilt udara fruit
·
To
extract the bioactive components of the fruit.
·
To
carry out photochemical analysis on the fruit extracts to determine
qualitatively and quantitatively the presence of these bioactive components
·
To
determine their MIC
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