Antibacterial Activity of Leaves and Fruits Extracts

1

Antibacterial Activity of Leaves and Fruits Extracts

Of the Gumbil Tree (Cordia africana L.)

Fadia SaifAldin Edrees Mohmmed

B. Sc. In Biology, Faculty of Education, University of

Holy Quran and Islamic Sciences, 2011.

Postgraduate Diploma in Biosciences and Biotechnology, Faculty

of Engineering and Technology, University of Gezira, 2013

A Dissertation

Submitted to the University of Gezira, in Partial Fulfillment of the

Requirement for the Award of the Degree of Master of science

in

Biosciences and Biotechnology (Biosciences)

Center of Biosciences and Biotechnology

Faculty of Engineering and Technology,

6/ 1/ 2015

Antibacterial Activity of Leaves and Fruits Extracts



2

Supervision Committee:

Name Position Signature

Prof. Awad Mohamed Abdel-Rahim Main Supervisor …………...

Dr. Yasir Mohamed Abdelrahim Co-supervisor …………...

Date of Examination: 6/ January /2015

Antibacterial Activity of the Extracts of the Leaves and Fruits



Examination Committee:

3

Name Position Signature

Prof. Awad Mohamed Abdel-Rahim Chairperson …………...

Prof. Ahmed Elawad Elfaki External Examiner …………...

Dr. Mai Abdalla Ali Abdalla Internal Examiner ………...

Date of Examination: 6/ January /2015

Dedication

To my

Dear father

Beloved mother

Brothers and sisters

(my grandmother)

Uncles and my aunts

Teachers

&

Friends

4

ACKNOWLEDGEMENTS

My grateful thanks to ALLAH who gave me the health and ability to

achieve this humble study, and with his will this achievement was

properly completed.

I would like also to express my thanks to all those who helped

me to carry out this work especially Professor Awad Mohamed

Abdel- Rahim, the main supervisor and Dr. Yasir Mohammed

Abdel-Rahim, co- supervisor and Dr. Mutaman Ali for assistance.

Thanks are also due the Technical Staff of the Microbiology

Laboratory, Faculty of Engineering and Technology, University of

Gezira.

Great thanks are due to my family and friends for support and

encouragement.

5

Antibacterial Activity of Leaves and Fruits Extracts of the Gumbil

Tree (Cordia africana L.)

Fadia Saif Aldean Edrees Mohmmed

M. Sc. In Biosciences and Biotechnology (Biosciences)

Center of Biosciences and Biotechnology

Faculty of Engineering and Technology

University of Gezira

ABSTRACT

Plant extracts were used to treat different diseases because they are

cheap and effective, but pharmacists are often reluctant to prescribe

them because of knowledge deficiency, real concerns about the

product safety, concerns about liability, and the presence of

pathogens contamination. The use of plants and their extracts as

remedies for curing many diseases have stimulated studies for

investigating the presence of effective antimicrobial substances in

them. The present study was investigating the antibacterial

properties of the tree Cordia africana (Gumbil) leaves and fruits

extracts. The inhibition zone and the total plate count methods were

used in this study. The results of the study showed that the extracts

were highly effective against the two tested bacteria. It inhibited

growth of the gram- negative (E. coli) bacterium. The extracts of

leaves with methanol, hexane, ethanol and water were effective

giving; 10, 10, 10 and 12 mm diameter, respectively. However, the

extracts of the fruit with ethanol and hexane were highly effective

both giving 12 mm compared methanol, chloroform and petroleum

6

ether which gave 10 mm. The water extract was the least (8 mm).

However, the effect of the fruit extracts was more serious against

Staph. aureus than on E. coli. Hexane and petroleum ether and

ethanol were highly effective (12, 11, 11 mm, diameter, respectively)

while methanol and chloroform gave intermediate effects (10 mm)

and the water was the least effective (7 mm).The inhibitory effect of

the water extract against the tested bacteria was more effective at the

higher concentrations of the extracts (75 and 100%,). The effect of

the water extract on the number of bacterial colonies was highly

pronounced. The number of E. coli was decreased from 226 colonies

at the control treatment to 29 colonies only at the higher

concentration. A similar was recorded for the Staph. aureus. It could

be recommended that, further tests should be carried on other

microorganisms, and the active antibacterial components need to be

verified.

7

شجشة انقبيم )أوساق وراسانشاؽ انؼاد نبكخيشيا نسخخهظاث Cordia africana L.)

فاديت سيف انذي ادسيس يحذ

اناجسخيش في انعهىو وانخقيت انبيىنىجيت )عهىو أحيائيت(

يشكز انعهىو وانخقيت انبيىنىجيت

كهيت انهذست وانخكىنىجيا

جايعت انجزيشة

انهخض

انباحاث في يعانجت انعذيذ ي حسخخذو يسخخهظاث

األيشاع ورنك ألها سخيظت وفعانت، غيش أ انخخظظي في يجال انظيذنت غيش يخحسي

السخخذايها ورنك نعذو انعشفت انخايت بها ونعذو انخأكذ ي ساليخها وقذسحها في انعانجت

حها نعانجت بعغ ووجىد انخهىد بانكائاث انشػت بها. اسخخذاو انباحاث ويسخخهظا

االيشاع حفزث انباحزي نذساست انىاد انفعانت انىجىدة بها كؼاداث نهكشوباث. حى في هزا

Cordia)انبحذ دساست انخىاص انزبـت نهبكخيشيا بسخخهظاث أوساق وراس شجشة انقبيم

africana) شث انخائج . حى اسخخذاو ؿشيقخي انـقت انزبـت وانعذد انكهي نهسخعشاث. أظه

ى انبكخيشيا أ نخهك انسخخهظاث فعانيت عانيت ػذ انبكخيشيا انخي حج دساسخها. فقذ ربــج

واناء أكزش (E. coli)انسانبت نظبغت جشاو . كاج يسخخهظاث االوساق بانيزاىل وانهكسي

س بااليزاىل يهى قـش بانخىاني . بيا كاج يسخخهظاث انزا 01,01,01,01فعانيت حيذ أعـج

يهى يقاست بسخخهظاث انيزاىل 01وانهكسي ها االكزش فعانيت حيذ اعـ كالها

يهى قـشا. وكا انسخخهض انائي هى االقم 01وانكهىسوفىسو وااليزش انبخشوني انخي اعـج

يهى(. هزا وكاج يسخخهظاث انزاس اكزشفعانيت عهي انبكخيشيا4فعانيت ) (Staph. aureus )

. وكاج يسخخهظاث انهكسي وااليزش انبخشوني وااليزاىل أكزش E. coli يقاست بانبكخيشيا

يهى عهي انخىاني(. بيا أعـج 00.00.01) Staph. aureusفعانيت ػذ انبكخيشيا

يهى( وكا انسخخهض انائي هى 01يسخخهظاث انيزاىل وانكهىسوفىسو حأريشاث يخىسـت )

(. كا انخاريش انائي انزبؾ نهسخخهض انائي أكزش فعانيت عه انخشكيزاث يهى3األقم فعانيت )

%( . كا حأريش انسخخهض انائي عهي حعذاد انسخعشاث انبكخيشيت أكزش 011,31انعانيت )

يسخعشة فقؾ 15يسخعشة في حجشبت انقاست اني 112وػىحا. حيذ اخفغ انخعذاد ي

ئج يشابهت في حسجيهها نبكخيشيا عه انخشكيز انعاني. انخا Staph. aureus حىطي انذساست .

8

بإجشاء انزيذ ي االخخباساث عه كائاث حيت دقيقت أخشي وكزنك نهخعشف عه انكىاث

. انفعانت ػذ انبكخيشيا

List of Contents

------

-

Topic

------------------------------------------------------------------

-----------

Main page

Supervision committee

Examination committee

Page

No.

I

II

III

Dedication Iv

Acknowledgements V

Abstract (English) VI

Abstract (Arabic) VII

List of Contents VIII

9

List of Tables X

List of Figures Xi

CHAPTER ONE

INTRODUCTION

Introduction 1

Objectives 4

CHAPTER TWO

LITERATURE REVIEW

2.1. Introduction 5

2.2. Plant investigated 8

2.2.1

2.2.2

2.2.3

2.2.4

2.2.5

2.2.6

2.2.7

2.2.8

2.3.

2.4

2.4.1

2.4.1.1

2.4.1.2

Local names

Geo graphic distribution

Tree management

Taxonomy and nomenclature

Botanical Description

Products and services

Biology

Ecology

Antimicrobial activity of plant extracts

Bacteria

Bacteria under test

E. coli

Staphylococcus aureus

8

8

9

9

9

10

10

11

11

13

13

12

CHAPTER THREE

MATERIALS AND METHODS

3.1. Sample collection 14

3.2 Source of cultures 14

3.3. Media used 14

11

3.3.1

3.3.1.1

3.3.1.2

Bacteriological media

Nutrient Agar

Nutrient Broth

14

14

14

3.4. Methods Used 15

3.4.1 The Cup-Plate Agar Diffusion Method 15

3.4.2 The Dilution Plate Method 15

3.5. Statistical Analysis 15

CHAPTER FOUR

RESULTS

4.1. Effect of Different Extracting Solvents 16

4.2.

4.3

Effect of Different Concentrations of the Aqueous

Extracts on the Inhibition Zone

Effect of Different Concentrations of the Extracts

on the Number of Colonies

21

21

CHAPTER FIVE

DISCUSSION

30

CHAPTOR SIX

CONCLUSIONS AND RECOMMENDATIONS

5.1 Conclusions 33

5.2 Recommendations 33

REFERENCES 34

11

List of Tables

Table

No.

Title Page

No.

4.1 Effect of different plant parts extracts obtained by

different solvents on the inhibition zone (mm) of E. coli

17

4.2. Effect of different plant parts extracts obtained by

different solvents on the inhibition zone (mm) of Staph.

aureus

19

4.3. Effect of different concentrations of Cordia africana

extracts on the inhibition zone of (mm) E. coli

22

4.4. Effect of different concentrations of Cordia africana 24

12

extracts on the inhibition zone (mm) of Staph. aureus


(leaves) extracts on the number of colonies of E. coli

26


(leaves) extracts on the number of colonies of Staph.

aureus

28

List of Figures

Figure

No.

Title Page

No

4.1 Effect of different plant parts extracts obtained by

different solvents on the inhibition zone of (mm) E.

coli

18

4.2. Effect of different plant parts extracts obtained by

different solvents on the inhibition zone of (mm)

Staph. aureus

20

4.3. Effect of different concentrations of Cordia africana 23

13

extracts on the inhibition zone (mm) of E. coli


extracts on the inhibition zone of (mm) Staph. aureus

25


leaves extracts on the number of colonies of E. coli

27


leaves extracts on the number of colonies of Staph.

aureus

29

INTRODUCTION

Natural products from plants have played major sustaining roles in the life of

humans, especially for food sources and for medicinal products. Nature has provided

mankind with folk medicines for centuries and continues to be the richest source of

bioactive chemicals for the development of modern drugs.

Herbalism is a traditional medicinal or folk medicine practice, based on the

use of plants and plant extracts. Herbalism is also known as botanical medicine,

medical herbalism, herbal medicine, herbolog, and phytotherapy. The scope of herbal

medicine is sometimes extended to include fungal and bee products, as well as

minerals, shells and certain animal parts.

Pharmacognosy is the study of medicines derived from natural sources

(Acharya and Shrivastava, 2008). Herbs had been used by all culture throughout

history. It was an integral part of the development of modern civilization. A herb is a

plant or a plant part used for its scent, flavor or throughout properties. Herbal

medicine products are dietary supplements that people take to improve their health.

Many herbs have been used for a long time for claimed health benefits. They are sold

as tablets, capsules, powders, teas, extracts of fresh or dried plants. However, some

can cause health problems, some are not effective and some many interact with other

drugs taken by the patient (Internet, 2010).

14

Clinical microbiologists have two reasons to be interested in the topic of

antimicrobial plant extracts. First, it is very likely that these photochemical

compounds, will find their way into the arsenal of antimicrobial drugs prescribed by

physicians; several are already being tested in humans. It is reported that, on average,

two or three antibiotics derived from microorganisms are launched each year (Clark,

1996). New sources, especially plant sources, are also being investigated. Second, the

public is becoming increasingly aware of problems with the over prescription and

misuse of traditional antibiotics. In addition, many people are interested in having

more autonomy over their medical care (Alper, 1998).

Worldwide spending on finding new anti-infective agents (including vaccines) is

expected to increase 60% from the spending levels (Alper, 1998). New sources,

especially plant sources, are also being investigated. Second, the public is becoming

increasingly aware of problems with the over prescription and misuse of traditional

antibiotics. In addition, many people are interested in having more autonomy over

their medical care. A multitude of plant compounds (often of unreliable purity) is

readily available over-the –counter from herbal suppliers and natural-food stores, and

self-medication with these substances is common place. The use of plant extracts, as

well as other alternative forms of medical treatment, is enjoying great popularity in

the late 1990s. Earlier in this decade, approximately one-third of people surveyed in

the United States used at least one "unconventional" therapy during the previous year

(Eisenberg et al., 1993).

About 25 percent of the prescription drugs dispensed in the United States

contain at least one active ingredient derived from plant material. Some are made

from plant extracts; others are synthesized to mimic a natural plant compound

(Internet, 2008).

The World Health Organization (WHO, 2002) estimates that 4 billion people

(80% of the world population) presently use herbal medicine for some aspects of

primary health care. Herbal medicine is a major component in all of the traditional

medicine. WHO (2002) also stated that out of the 119 plant-derived pharmaceutical

medicine, about 74 percent was used in modern medicine in ways that correlated

directly with their traditional uses as plant medicine by native cultures (Internet,

2008).

Substances derived from the plants, remain the basis for a large proportion of

commercial medications used today for treatment of heart diseases, high blood

15

pressure, pain, asthma, and other health problems. For example; the use of clove plant

in modern medicine. The herb has been in use since 1775. At present, the powder

leaves of the plant are known as the cardiac stimulant digitalis to the millions of heart

patients, it keeps them alive worldwide (Internet, 2008).

Traditional use of medicines is recognized as a way to learn about the

potential future medicines. In 2001, researchers identified 122 compounds used in

main stream medicine which were derived from "ethno medical" plant sources; 80%

of these compounds were used in the same or related manner as the traditional ethno

medical use (Fabricant and Farnsworth, 2001).

Many plants synthesize substances that are useful to the maintenance of health

in humans and animals. These include aromatic substances, most of which are phenols

or their oxygen-substituted derivatives such as tannins. In many cases, these

substances serve as plant defense mechanisms against predation by microorganisms,

insects, and herbivores.

Finding healing powers in plant is an ancient idea. People on all continents

have long time applied poultices and imbibed infusions of hundreds, if not thousands,

of indigenous plants, dating back to prehistory. There is evidence that Neanderthals

living 60,000 years used plants such as holly hock for traditional medicine (Stockwell,

1988 and Thomson, 1978).

These plants are still widely used in ethno medicine around the world.

Historically, therapeutic results have been mixed; quite often cured or symptom relief

resulted. Poisonings occurred at a high rate, also. Currently, of the one-quarter to one

half of all pharmaceuticals dispensed in the United States having higher-plant origins,

very few are intended for use as antimicrobials.

Gumbil (Cordia africana) is a wide spread tree in tropical Africa. It is

spreading, from Guinea east to Eritrea, Ethiopia and Kenya, and south to Angola,

Zimbabwe, Mozambique and northern South Africa. It is also found in Saudi Arabia

and Yemen, and has been planted in many tropical countries. In Sudan the tree is

confined to the areas of Ad- Damazin, Dafror (Jabal Merra) and Kordofan

(Drummond, 1981).

Generally, Cordia species (Boraginaceae) contain flavonoids (Sertie et al.,

1990) polyphenols (Marston et al., 1988) anti-androgenic triterpenoids, saponins,

sesquiterpenes and cromenes (Dos Santos et al., 2005). Antifungal and larvicidal

16

properties of Cordia sp. have been referred to the presence of heteroterpenoids,

naphthoquinones and naphthoxirene (Loset et al., 1998).

In the Sudanese traditional medicine, Cordia africana (Gumbil) is widely

known as a treatment for liver disease, fever and intestinal complaints such as

dysentery (Burkill, 1985). The polysaccharide from the fruit pulp of Cordia africana

consists mainly of galactose, mannose, xylose, arabinose, glucose, rhamanose,

galacturinc acid and pharmaceutic acid and about 2.5% protein. It may have

applications in food, textile and pharmaceutical industries and could act as an

emulsifying agent also in systems with relative high salt concentration. Moreover, the

wood is moderately durable, and moderately resistant to termite and pinhole borer

attacks (Bekele_ Tesemm, 2007).

Objectives of the study

General objective:

The overall aim of this study was to find environmentally friendly

antibacterial compounds against two bacteria (E. coli and Staph. aureus).

Specific objectives:

1. Different solvents will be tested as extracting agents.

2. The study has been made to investigate the antibacterial activity of the

extracts from Cordia africana (Gumbil).

3. The extracts from the different parts of the tree will be examined for their activity against the two bacteria (E. coli and Staph. aureus) using different concentrations.

17

CHAPTER TWO

LITERATURE REVIEW

2.1 Introduction A forest (also called a wood or woods) is best defined as an ecosystem or an

assemblage of ecosystems dominated by trees and other woody vegetations. Today,

there are more than 250 definitions of the term "forest." These definitions differ

basically, on the emphases or concerns of different people. The total world forested

area is approximately 3.9 Billion hectares: this represents 30% of the land surface area

(Lund, 2006(.

Forest plays a crucial role in providing a wide range of products, including

food and fruits, fodder for livestock (Campbell and Tewaria1995; CIFOR, 2005), fuel

wood, charcoal and timber, which remains the basic forest product even in the highly

technological society. The livelihood of the majority of rural people depends on the

forest as prime sources of fuel wood as well as for non timber forest product such as

fruits, fibers and medicine (Barrow et al., 2002; Campbell, 1986; Falconer 1990; FAO

1983 ( .

18

Non wood forest products )NWFP), play a vital role in the livelihood of people

in and around the forests (Grimes et al., 1994). Innovative forest management could

be efficient in providing a wide variety of active compounds with diverse structures

and biological activities. Forests contain different phytochemical compounds as

natural products, for example Flavonoids (Yang et al., 1990( and sesquiterpenes

which were isolated from Cleome droserifolia, family Ceomaceae. (Hussein et al.,

1994). Quinine, a known alkaloid, was isolated from the cinchona genus of trees,

located mainly is South America (Brogger and Kharazmi, 2011). Taxol (paclitaxe), a

cytotoxic diterpen alkaloid, was isolated from the park of the pacific yew tree Taxys

brevifolia in the late 1960s (Wall et al., 1966). The most known and most important

Sudanese NWFP is the Gum Arabic, obtained from Acacia Senegal (hashab), but also

from Acacia seyal (talha). Sudan is also one of the world‘s biggest export of olibanum

resins from the tree (Bosswellia sp( .

Recently, plant possessing insecticidal substances has generated extraordinary

interest as potential source of natural insect control agent (Murray, 2006). The

concept of pesticide, now, has a profound change and does not stress certainly

"killings”, but rather "regulation of the pests". It is though that in the 21th century,

pesticides should be called environment acceptable pesticide or environment friendly

pesticide (Li and Li, 2007; Xu et al., 2005; Wu, 2004). Therefore, in the process of

searching for a safe replacer of the chemically synthetic pesticides, plant source

pesticides have become alternative because plant source pesticides have the advantage

of reducing harmful particle quantity entering into the environment, being safe to

human and livestock, decomposing quickly in nature, being not inferior to the

chemically synthetic pesticides in the toxicity aspect, and being not easy for the

pathogen and the pest to have resistance (Yu et al., 2001) .

Other vegetal NWFP are fodder (e.g. Ziziphus sp., Acacia sp.): food (fruits, e.g.

Hyphaene thebaica;nuts, e.g. Cordeauxia edulis; and edible oils, e.g. Vitellaria

paradoxa, Balanites aegyptica); medicines (e.g.Tamarindus indica ); dyes (e.g Henna

Lawsoniainermis, Prosopis africana): fibers (e.g.Borassus aethiopum); latex (e.g.

Landolfia ovariensis); and tannins (e.g.Rizophora mucronate, Acacia nilotica).

Honey and bees wax are the only faunal NWFP on which documentation exists.

Altogether, about 150 NWFP are used in Sudan (Badi, 1993). Plant products are of

major importance, especially for subsistence (e.g food, medicines and fodder). At

least six NWFP are exports: Gum Arabic (Acacia sp.), gum loban )Boswellia

19

papyrifera, Commiphora sp.), as well as, the fruits of aradabe (Tamarindus indica).

The total value of foreign trade amounts to US$6929587 (Sulieman and Eldoma,

1994).

Much of the medicinal use of plants seems to have been developed through

observations of wild animals, and by trial. Indeed, well into the 20th century much of

the pharmacopoeia of scientific medicine was derived from the herbal lore of native

people. Mainstream medicine is increasingly receptive to the use of antimicrobial and

other drugs derived from plants, as traditional antibiotic (products of microorganisms

or their synthesized derivatives), become these have been ineffective and as new,

particularly viral diseases, remain intractable to this type of drugs. Another driving

factor for the renewed interest in plant antimicrobial in the past 20 years has been the

rapid rate of (plant) species extinction (Lewis and Elvin, 1995). Records from the

ancient Egyptians and Chinese showed that plants were used for the preparation of

hundreds of drugs, covering a most impressive array of health problems and diseases

(Osbourn and Lanzotti, 2009).

Natural products have, until recently, been the primary source of commercial

medicines and drug leads. A recent survey showed that 61% of the 877 drugs

introduced worldwide can be traced to or were inspired by natural products. However,

beginning in the 1990s natural product drug discovery was virtually eliminated in

most big pharmaceutical companies. This was primarily due to the promise of the

then-emerging field of combinatorial chemistry (Cseke et al., 2004), whereby huge

libraries of man-made small molecules could be rapidly synthesized and evaluated as

drug candidates. However, from 1981 to 2002, no combinatorial compounds became

approved as drugs, although several are currently in late-stage clinical trials. At the

same time, the number of new drug pharmaceutical corporations is painfully aware.

The haystack is larger, but the needle within it is more elusive. This has led only

recently to a new found respect for the privileged structures inherent within natural

products (De Simone et al., 2004). Of the roughly 350,000 species of plants believed

to exist, one-third reported, only a fraction of them have been chemically investigated.

Many countries have developed systems for exploration as well as preservation. At

the same time, habitat loss is the greatest immediate threat to biodiversity within their

borders and has developed systems for exploration as well as preservation. At the

same time, habitat loss is the greatest immediate threat to biodiversity (Frankel et al.,

1995). Plants produce an amazing array of organic chemicals with an enormous

21

diversity of structural types. Many of these phytochemicals are essential for plant

growth and development and are widely used by humans and other animals as food

sources. They include a wide variety of polysaccharides such as cellulose, starch and

fructans, the polyphenols, lignin, fatty acids and lipids, proteins such as enzymes and

structural components in cell membranes; and nucleic acids such as DNA and RNA.

Many more have undoubtedly evolved in response to ecological pressures of

competition, including plant-to-plant competition for light and space, herbivory from

marauding insects and other fauna, as well as bacterial and fungal infections (e.g.

phytoalexins). These biological active compounds are not only necessary for the well-

being, survival, and evolution of the plants that produce them, but also for humans,

who have exploited them for industrial processes (e.g. ethanol from corn and

sugarcane as an alternative energy source and, currently, pharmaceutical

biotechnology and nanotechnology), S-methiyltcytosine vicine convicine theobromine

zeatin construction (e.g. houses, bridges, barrels, baseball, bats, fences, insulation),

fuel (e.g., crop residues, wood chips, and sawdust), agricultural (e.g., fruits, vegetable,

herbs and spices, wine and beer products, animal feeds, forest and horticultural

products, and landscape plants), medical/pharmaceutical, religious purposes,

recreational, and even spiritual/ religious purposes (Leland et al., 2006). There is an

increasing demand for biologically active research approach. The synthetic chemical

pharmaceuticals showed various side effects on the functioning of different parts of

the body, both internally and therapeutic potential, due to the presence of active

pharmacologically important substances, such as terpenes, alkaloids, flavonoids and

glycosides (Yusuf et al., 2002 and Farrukh and Ahmed, 2003). Screening is a tool in

discovering new biologically active molecules which have been found to be most

productive in the area of antimicrobial activity in India and abroad (Grierson and

Afolayan, 1999 and Karuppusamy et al., 2001).

2.2 The Plant Investigated (Gumbail)

2.2.1 Local Names

In Afrikaans it is called (grootblaarpieringbessie); in Amharic (wanza); in

Arabic (gambil); in English (Sudan teak), in East African (cordia, large-leafed

cordia); in French (sebestierd' Afrique); in Yuganda (mukebu); in Swahili

(mringaringa, mringamringa, mukumari, makobokobo); in Tigrigna (auhi, ekhi,

21

awhi); and its Trade name is mukebu, mukumari (Orwa. et al., 2009). It has the

scientific name (Cordia africana Lam).

The tree is a deciduous tree up to 30 m tall, characterized by a very dense

foliage consisting of large, dark green, cordial leaves (Bekele-Tesemma, et al., 1993).

It is a large shady tree spreading for 10 m. It is sometimes named Cordia abyssinica

and the local name in Sudan is "gumbail" (Bein, 1996).

2.2.2 Geographic Distribution

The origin Cordia Africana is Africa. It is widely spreaded in tropical Africa. It

is spreading, from Guinea east to Eritrea, Ethiopia and Kenya, and south to Angola,

Zimbabwe, Mozambique and northern South Africa. It is also found in Saudi Arabia

and Yemen, and has been planted in many tropical countries.

In Sudan the tree is confined to the areas of Ad- Damazin, Dafror (Jabal Merra)

and Kordofan (Drummond, 1981).

2.2.3 Tree Management

The species grows fairly fast, reaching 7-8 m in 7 years; management

practices include pollarding, lopping and coppicing. After extraction, seeds are dried

in the sun to 6-8% mc; can be stored for at least 1 year in hermetic storage at 3 C°.

with no loss in viability. There are about 18 000 seeds/kg (Orwa et al., 2009).

2.2.4 Taxonomy and Nomenclature

Kingdom: Plantae

Class: Angiospermae

Subclass: Eudicots

Superorder: Asteridae

Order: Boraginales

Family: Boraginaceae

Subfamily: Cordioideae

Genus: Cordia

Species: africana

Scientific name: Cordia africana (Quattrocchi, 2000).

22

2.2.5 Botanical Description

A small tree 6-15 m high. Bole small much-branched, typically curved or

crooked. Bark pale brown, shallowly fissured. Leaves alternate, ovate to suborbicular

7.5- 17 x 2.5 – 7.5 cm, glabrous above shortly tomentose beneath, petioles 3- 5cm

long, glabrous. Inflorescence shortly pedicellate or subsessile, massed in compact

panicles covering the crown of trees with a mass of white flowers; sepals strongly

ribbed, about 1 cm long, brown tomentellous; petals white, long exerted, funnel-

shaped, about 2.5 cm long, lobes short, rounded; stamens 5, epipetalous; style 4- fid.

Fruit is brown ovoid drupes about 1.25 cm diameter with persistent calyx at base

(Hamza, 1990).

2.2.6 Products and Services

Food: Mature fruits have a sweet, mucilaginous, edible pulp.

Fodder: Leaves provide fodder for the dry season. Apiculture: C. africana provides

good bee forage, as the flowers yield plenty of nectar. Beehives are often

placed in the trees.

Fuel: Trees are a good source of firewood.

Timber: The heartwood is pinkish-brown, reasonably durable, relatively termite

resistant; it works easily and polishes well but is often twisted and difficult

to saw. It is used for high-quality furniture, doors, windows, cabinet

making, drums, beehives, joinery, interior construction, mortars, paneling

and veneering.

Medicine: The fresh, juicy bark is used to tie a broken bone; this splint is changed

occasionally with a fresh one until the bone is healed.

Shade or shelter: C. africana is planted as a shade tree in coffee plantations; it is

usually left in the fields, as it provides excellent shade for crops.

Soil improver: Leaf fall in the dry season is heavy, and the leaves make good mulch.

Ornamental: Trees are planted in amenity areas (Orwa et al., 2009).

23

2.2.7 Biology

Flowering starts when the trees are 3-5 years old. In Sudan, flowering occurs in

October to December and fruiting from January to April; in Kenya, flowering is from

April to June. It is repeated at intervals over several weeks and is evidently triggered

of by rain showers. After pollination by insects, fruit development takes a period of

almost 6 months. Fruit is eaten and probably dispersed by birds (Orwa et al., 2009).

2.2.8 Ecology

The species occurs at medium to low altitudes, in woodland, savannah and bush,

in warm and moist areas, often along river banks. It will grow in drier conditions but

thrives in good rainfall areas and is scattered in occurrence. It occurs in afro-montane

rainforest and undifferentiated afro-montane forest (mixed Podocarpus forest), usually

along margins and in clearings. It is an early colonizer in forest regrowth. It is often

left when forests are cleared for cultivation, as the tree is an excellent shade tree for

crops. Also found in riverine forest and secondary bushland, transgressing into humid

types of woodland. In West Africa, this species seems to be restricted to montane and

submontane habitats; it has limited distribution in the lowland habitats of the

Democratic Republic of Congo (Orwa et al., 2009).

2.3 Antimicrobial Activity of Plant Extracts

Plants are potent biochemical factories and have been components of

phytomedicine since times immemorial. Man is able to obtain from them a wondrous

assortment of industrial chemicals. Plants- based natural constituents can be derived

from any part of the plant, like bark, leaves, flowers, roots, fruits, seeds, etc. and any

part of the plant may contain active components. The beneficial medicinal effects of

plant materials are typically result from the combinations of secondary products

present in the plant. The medicinal actions of plants are unique to particular plant

species or groups and are consistent with the concept that the combination of

secondary products in a particular plant is taxonomically distinct (Wink, 1999).

Medicinal plants have been used for centuries as remedies for human diseases because

they contain components of therapeutic value. Plants containing near infrared (NIR)

spectroscopy, were used to classify insect- infested and sound seeds of a tropical

24

multipurpose tree, Cordia africana Lam. A calibration model derived by partial least

squares regression of orthogonal signal corrected spectra resulted in a 100%

classification rate. Difference spectrum and partial least squares weight indicated that

absorbance differences between insect–infested and sound seeds might have been due

to differences in components as well as moisture content. The results showed the

possibility of using NIR spectroscopy in the seed cleaning process in the future,

provided that appropriate sorting instruments are developed. The acceptance of

traditional medicine as an alternative form of health care and the development of

microbial resistance to the available antibiotics has led researchers to investigate the

antimicrobial activity of medicinal plants (Hostettmann and Nakanishi , 1979).

2.4 Bacteria

Many of us know bacteria only as "germs," invisible creatures that can invade

our bodies and make us sick. Few know that many bacteria not only coexist with us

all the time, but help us do an amazing array of useful things like the production of

vitamins, the break down of some garbage, and even the maintenance of our

atmosphere. Bacteria consist of only a single cell, but don’t let their small size and

seeming simplicity fool you. They are an amazingly complex and fascinating group of

creatures. Bacteria have been found to live in temperatures above the boiling point

and in cold that would freeze your blood. They "eat" everything from sugar and starch

to sunlight, sulfur and iron. There is even a species of bacteria, Deinococcus

radiodurans, that can withstand blasts of radiation 1,000 times greater than would kill

a human being (American Society for Microbiology, 2014).

There are thousands of species of bacteria, but all of them are basically one of

three different shapes. Some are rod or stick-shaped and called bacilli. Others are

shaped like little balls and called cocci others still are helical or spiral in shape. Some

bacteria cells exist as individuals while others cluster together to form pairs, chains,

squares or other groupings.

Bacteria fall into a category of life called the prokaryotes (pro-carry-oats).

Prokaryotes genetic material, or DNA, IS not enclosed in a cellular compartment

called the nucleus. Bacteria and archaea are the only prokaryotes. All other life forms

are Eukaryotes (you-carry- oats), creatures whose cells have nuclei.

Bacteria are used in industry in a number of ways that generally exploit their

natural metabolic capabilities. They are used in manufacture of foods and production

25

of antibiotics, drugs, vaccines, starter cultures, insecticides, enzymes, fuels and

solvents. In addition, with genetic engineering technology, bacteria can be

programmed to make various substances used in food science, agriculture and

medicine. The genetic systems of bacteria are the foundation of the biotechnology

industry discussed below (Internet, 2008).

In industry the pharmaceutical, lactic acid bacteria, such as Lactobacillus,

Lactococcus and Streptococcus are used in the manufacture of dairy products such as

cheeses, including cottage cheese and cream cheese, cultured butter, sour cream,

buttermilk, yogurt and kefir. In the wine making process, when the yeast cells are kept

short of oxygen, they cause sugars in grape juice to ferment, forming alcohol and

carbon dioxide. In the making of cheese and yoghurt, bacteria perform a similar job,

turning lactose into lactic acid. Rennet is then added to separate the curds and whey.

2.4.1 Bacteria under Test

2.4.1.1 E. coli

E. coli is a Gram negative rod-shaped bacterium that is commonly found in the

lower intestine. Most E. coli strains are harmless. But some can cause serious humans

food poisoning (Vogt and Dippold, 2005). The harmless strains are part of the normal

flora of the gut (Bentley and Meganthan, 1982).

E. coli are not always confined to the intestine and their ability to survive for

brief periods outside the body makes them an ideal indicator organisms to test

environment samples for fecal contamination. The bacterium can also be grown easily

and its genetics are comparatively simple and easily manipulated or duplicated trough

a process of mutagenesis, making it one of the best- studied prokaryotic model

organisms, and an important species in biotechnology and microbiology.

E. coli was discovered by the German pediatrician and bacteriologist Theodor

Escherich in 1885, and is now classified as part of the Enterobacteriaceae family of

gamma-proteobacteria (Feng et al., 2002).

2.4.1.2 Staphylococcus aureus

26

The Gram- positive cocci forms a heterogeneous collection of

approximately 16 genera that colonize human. Features that they have in common are

their spherical shape, their Gram strain reaction and the absence of endspores. There

are two groups of bacteria, catalase-positive genera (Staphylococcus, Micrococcus

and Stomatococcus) and catalase-negative genera (Streptococcus. Entreococcus and

related organisms) as described by Murray et al., (1998).

Staphylococcus aureus and related organisms are generally wide spread in

nature, their normal habitats being the skin, skin glands and the mucus membranes of

humans and other mammals (Devries, 1986).

27

CHAPTER THREE

MATERIALS AND METHODS

3.1 Sample Collection

Sample of different parts of the tree (Cordial africana) fruits and leaves were

obtained from Darfor and were collected in June 2014.

Aqueous extracts of the powder (50 - 500g/ml) of the seeds and leaves were added

to the different media used, to give different concentration (0.00, 25.00, 50.00, 75.00

and 100 mg/ml)

3.2 Source of Cultures

Two bacterial isolates (E. coli and Staphylococcus aureus) were obtained from

the laboratory in the Department of Food Science and Technology, University of

Gezira, Wad Medani, Sudan.

3.3 Media used

3.3.1 Bacteriological Media

3.3.1.1 Nutrient Agar

The medium composition is as follows :(g/l)

Lap-lemco powder 1.0

Yeast extract 2.0

Peptone 5.0

Sodium chloride 5.0

Agar 15.0

Twenty- eight grams of an already prepared medium (Himedia Ltd.), were used

per one liter (1000 ml) of distilled water. The medium was dispensed into flask (250

ml), and autoclaved at 121 Cº (151b/in2) for 15 minutes, then poured into sterile Petri

dishes, which were allowed to solidity and Kept inverted into a refrigerator before

use.

3.3.1.2 Nutrient Broth

The medium composition as above except that agar was not added.

28

3.4 Methods Used

3.4.1 The Cup-Plate Agar Diffusion (Inhibition Zone) Method

This method was used, using Nutrient Agar (N.A) in this method two ml of

standardized bacterial cell suspension (10×105) of E. coli or of Staphylococcus aureus

were thoroughly mixed with 200 ml of sterile molten nutrient agar. The medium was

then distributed into sterile Petri-dishes and left to solidify at room temperature for 24

hours. Sterile Whatman glass fiber discs (No.5) were saturated with the extract of

Cordia africana, then allowed to dry and transferred centrally on the surface of the

solidified medium in each plate. The plate was then incubated at room temperature for

72hours and the inhibition zones were measured as described by Barry et al. (1970)

and Cruickshank et al. (1975). Three replicates were made for each treatment.

3.4.2 The Dilution Plate Method

The effects of the aqueous extracts of Cordia africana on bacterial growth were

assessed by the Dilution Plate Method. Nutrient broth medium batches containing

different concentration of the Cordia africana extract were distributed into

McCartney bottles and each was inoculated by one ml of the bacterial suspension

prepared as above inoculated bottles (three per treatment) were incubated at room

temperature and the bacterial growth was measured by the dilution plate method, were

1ml was drawn from each bottle and placed in a tube containing 9 ml sterile distilled

water and serially diluted.

From each dilution 0.1 ml was removed by a sterile pipette and placed

on the surface of an already solidified Nutrient Agar (N.A) in Petri- dishes and

spreader on that surface, using a sterile glass rod. Inoculated plates were kept (28- 30

C°) and the number of colonies was counted at different intervals of time. The number

was calculated as a log number of colonies per one ml. The logs of the numbers were

plotted against time.

3.5 Statistical Analysis

The statistical analysis methods used in the work was ANOVA and Duncan‘s

MRT.

29

CHAPTER FOUR

RESULTS

4.1 Effect of Different Extracting Solvents

The present study investigated the antibacterial activity of the extracts of Cordia

africana (Gumbil) plant using two bacteria (E. coli and staph. aureus). Different

extracting solvents were used including; chloroform, ethanol, methanol, petroleum

ether, as well as water (aqueous extract). The cup plate inhibition zone method was

used for the comparison between the different solvents. Results in Table (4.1.) and

Fig. (4.1.) are showing the effects of the extracts obtained by the different solvents on

the inhibition zone of the bacterium, E. coli.

From the results it is clear that generally, ethanol and hexane were better than

the others. These were followed by methanol, petroleum ether, water, and then

chloroform. However, water extracts were also effective compared to the chloroform

treatment. The results also showed that the different plant parts were showing

different antibacterial effects when extracted with different solvents. The fruits gave

better effects when extracted with hexane and ethanol, while the leaves were highly

effective when extracted with methanol. Petroleum ether and chloroform leaf extracts

as well as the water extract of fruits, the were least in inhibiting E. coli growth.

Intermediate effects were shown by the extracts of leaves with both ethanol and

hexane as well as with methanol extract of the fruits.

On the other hand, Table (4.2) and Fig (4.2) are showing the effects of the

extracts obtained by the different solvents on the inhibition zone of the bacterium,

Staph. aureus. The results indicated that generally, the water extracts of the leaves

were better than the all other extracts. The results also confirmed that the different

plant parts were showing different antibacterial effects when extracted with different

solvents. However, the water extracts of the fruits and the chloroform extracts of the

leaves were the least effective. The fruits gave better effects when extracted with all

solvents except with water, while the leaves were only highly effective when

extracted with water and ethanol. Petroleum ether, methanol, chloroform and hexane

leaf extracts, were less effective in inhibiting E. coli growth.

31

Table (4.1): Effect of different plant parts extracts obtained by different solvents

on inhibition zone (mm) of E. coli

Solvents Plant parts

Mean Leaf Fruit

Chloroform 6 10 8

Ethanol 10 12 11

Hexane 10 12 11

Methanol 12 10 9

P. ether 8 10 9

Water 10 8 9

Mean 9.33 10.33

Anova table

S E S D F Fcrit

Leaf 0.84 2.57 Row 1.38 6.39

Fruit 0.61 1.5 Colum 6.62 7.71

31

Figure (4.1): Effect of different plant parts extracts obtained by different

solvents on inhibition zone (mm) of E. coli

32

Table (4.2): Effect of different plant parts extracts obtained by different solvents

on the inhibition zone (mm) of Staph. aureus

Anova table

Solvents Plant parts Mean

Leaf Fruit

Chloroform 6 10 8

Ethanol 10 11 10.5

Hexane 8 12 10

Methanol 8 10 9

P.ether 8 11 9.5

Water 18 7 12.5

Mean 9.67 10.17

S E S D F Fcrit

Leaf 1.75 4.27 Row 0.28 5.50

Fruit 0.703 1.72 Colum 0.045 6.61

33

Figure (4.2): Effect of different plant part extracts obtained by different solvents

on inhibition zone of (mm) Staph. aureus

34

4.2 Effect of Different Concentrations of the Aqueous Extracts on

the Inhibition Zone

The effect of the different concentrations of the aqueous extracts plant parts

(leaves and fruits) on inhibition zone of both bacteria (E. coli and Staph. aureus) were

also investigated in the present study. The inhibition zone method described above

was also for the present study.

The results of the effects on the different concentrations of the extracts of the

different plant parts on E. coli are shown in Table (4. 3) and on Fig. (4. 3). The results

showed that there is an increasing effect with the increasing concentration of the

extract reaching its maximum at the concentration 100%. However, with the fruit

extracts both concentration of 25 and 50% were giving a similar effect (6 mm), while

the other two concentrations 75 and 100% were also similarly effective (8 mm).

The effects of the different concentrations of the aqueous plant parts (Leaves

and Fruits) on inhibition zone of bacterium Staph. aureus are shown in Table (4.4)

and Fig. (4.4). The results showed that there is an increasing effect with the increasing

concentration of the extract reaching its maximum at the concentration 75% and for

both extracts the inhibition zone was decreased at the maximum concentration

(100%). The maximum effect for both leaves and the fruits extracts at the

concentration 75% was 20 mm and 10 mm, respectively.

The effects of the different concentrations, on both E. coli and Staph. aureus,

were statistically significant compared to the control.

4.3 Effect of Different Concentrations of the Extracts on the Number

of Colonies

The effects of the extracts of leaves and fruits of Cordia africana on the number of

bacterial colonies were investigated using the dilution plate method described in the

materials and methods section.

The extracts were found highly effective in reducing the number of colonies of

both bacteria, E. coli (Table 4.5 and Fig. 4.5) and Staph. aureus (Table 4.6 and Fig.

4.6). The effect was much greater against Staph. aureus, the fact which indicated that

the bacterium Staph. aureus is more sensitive to the extracts compared to E. coli.

35

Table (4.3): Effect of different concentrations of Cordia africana extracts on the

inhibition zone of (mm) of E. coli.

Concentration %

Plant parts

Mean

Leaf Fruit

0 0 0 0

25 10 6 8

50 10 6 8

75 10 8 9

100 10 8 9

Mean 8 5.6

Anova table

F crit F SD SE

3.84 1.56 Row 4.47 2 Leaf

4.46 6.97 Colum 3.28 1.47 Fruit

36

Figure (4.3): Effect of different concentrations of Cordia africana

extracts on the inhibition zone (mm) of E. coli.

37

Table (4.4): Effect of different concentrations of Cordia africana extracts on

inhibition zone (mm) of Staph. aureus

Concentration %

Plant parts

Mean

Leaf Fruit

0 0 0 0

25 10 7 8.5

50 10 8 9

75 20 10 15

100 18 7 12.5

Mean 6.4 11.6

Anova table

Fcrit F SD SE

6.39 5.24 Row 1.69 3.54 Leaf

7.71 5.47 Colum 3.78 7.92 Fruit

38

Figure (4.4): Effect of different concentrations of Cordia africana extracts on

inhibition zone of Staph. aureus

39

Table (4.5) : Effect of different concentrations of Cordia africana (leaves) extract

on the number of colonies of E. coli

Concentration% Incubation Period (days)

Mean 2 4 6 8

0 171 183 208 226 197

25 49 60 73 93 68.75

50 35 45 51 67 49.5

75 20 27 35 51 39.25

100 9 17 24 29 19.75

Mean 56.8 66.4 78.2 93.2

Anova Table

Fcrit F SD SE Days

6.39 364.28 Row 34.80 33.56 6

7.71 9.45 Colum 77.82 75.05 8

41

Figure (4.5) : Effect of different concentrations of Cordia africana (leaves) extract

on the number of colonies of E. coli

41

Table (4.6) : Effect of different concentrations of Cordia africana (leaves) on the

number of colonies of Staph. aureus

Concentration

%

Incubation Periods (days)

Mean 2 4 6 8

0 114 121 136 148 129.75

25 45 57 61 73 59

50 34 45 46 56 45.25

75 19 31 39 44 33.25

100 7 18 25 29 19.75

Mean 43.8 54.4 61.4 70

Anova Table

Fcrit F SD SE Days

2.90 310.23 Row 34.67 19.53 6

3.28 17.49 Colum 46.49 20.79 8

42

Figure (4.6): Effect of different concentrations of Cordia africana (leaves) on

number of colonies of Staph. aureus

43

CHAPTER FIVE

DISCUSSION

The present study was investigating the biological activity of the extracts of

leaves and fruits of Gumbail (Cordia africana) against two bacteria (E. coli and

Staph. aureus). Two methods were employed; the cup-plate agar diffusion (inhibition

zone) method and the count plate agar (dilution plate) method. From the results it is

clear that the leaf and the fruit extract of Cordia africana using the inhibition zone

method gave inhibition zones of growth against the two bacteria (E. coli and Staph.

aureus) far greater than that of the control treatment. Results obtained by the count

plate method, showed that the number of colonies of both bacteria was highly

significantly reduced, even at the lower concentrations.

The results also indicated that the Cordia africana leaf and fruit extracts are

more effective against Staph. aureus, compared to the results on E. coli. Antibacterial

activity of plant extracts are well documented (Alicia, 1981). Vlietinck et al. (1995).

Screened about 100 medicinal plants used by traditional healers to treat infection in

Rwanda, for their antibacterial, antifungal and antiviral properties. Their study

showed that 45% of the plants were active against Staph. aureus, 2% against E. coli

16% against Pseudomonas aerogenosa and 7% against Candida albicans. Moreover,

about 27% of the plants tested exhibited antiviral properties. In Sumatra (Indonesia),

114 plant extracts were assayed for their antibacterial activity (Ahmed, 2002). About

82% of the extracts were active against Staph. aureus while 32% of them were active

against E. coli . However, the plant Garcinia cowa, which was reported to be rich in

xanthenes showed a moderate antibacterial activity against Staph. aureus (Pattalung

et al., 1994). According to Encamation et al. (1994), the flavones isolated from the

medicinal plant Culliandra colitornia exhibited an antibacterial activity against the

tow bacteria isolates tested. The extracts of plant leaves of Tagetes minuta were found

to exhibit some activity against both Gram positive and Gram negative bacteria

(Tereschuk et al., 1997).

In Sudan many studies were carried out for testing the antimicrobial activity of

some medicinal plants. Ahmed (2004) tested the extracts of 10 plants against Gram

positive and Gram negative bacteria as well as against Condida albicans. He found a

marked effect against the Gram positive Staph. aureus followed by E. coli and

Condida albicans, respectively. Plants may represent a potential source of antibiotics

44

as evidenced by the huge number of studies dealing with antimicrobial activities

(Aldar man 1976; Marth; Pratt, 1977; and Pelcazar et al., 1977). The use of plants and

their extracts as remedies for curing many diseases have stimulate studies for

investigating the presence of effective antimicrobial substances in them (Ahmed;

2002, Abdel Daim , 2001; Sulieman et al., 2008; Abdel-Rahim et al., 2010).

Clinical microbiology has two reasons to be interested in the antimicrobial

activity of plant extracts. First, it is very likely that these phytochemicals will find

their way into the arsenal of antimicrobial drugs prescribed by physicians; several are

already being tested in humans. It is reported that, on average, two or three antibiotics

derived from microorganisms are launched each year (Clark, 1996). New sources,

especially plant sources, are also being investigated. Second, the public is becoming

increasingly aware of the problems with the over prescription and misuse of

traditional antibiotics. In addition, many people are interested in having more

autonomy over their medical care (Alper, 1998).

Terrestrial plants in particular were used as the basis of sophisticated

traditional pharmacopeias form as early as 2600 BC, and some of the earliest

documentations come from inscriptions from Mesopotamia. Antimicrobial activity of

plant extract are well documented (Alicia, 1981). Vlietinck et al. (1995) and Odebiyi

and Sofowra (1979) screened about 100 medicinal plants used by traditional hearlers

to treat infection in Rwanda, Nigeria and Sudan for their antibacterial, antifungal and

antiviral properties. Roodt (1992) reported antibacterial activity of kigelia africana

bark extract (aqueous, methanol and chloroform) against Escherchia coli,

Enterobacter aerogens, Klebsiella pneumonia, Salmonella typhi, proteus vulgaries,

Pseudomonas aeruginosa, Staphylococcus aureus and Baceillus cereus. Also he

showed that the methanolic bark extracts were highly effective against Salmonella

typhi and Proteus vulgari. In recent years, a significant revival of interest in the

natural products as a potential source for new medicines has been observed among

academia as well as pharmaceutical companies. Several modern drugs (40% of the

modern drugs in use) have been developed from natural products. More precisely,

39% of the 520 new approved drugs between 1983 and 1994 were natural products or

their derivatives, and 60-80% of the antibacterial and the anticancer drugs were from

natural origins. In 2000, approximately 60% of all drugs in clinical trials for the

multiplicity of cancers had natural origins (Cragg et al., 1997). Plants contain

significant levels of natural genetic and phenotypic variation between individuals

45

within a species for traits ranging from development to metabolism to pathogen

resistance. Natural products have been a source of therapeutic agents for thousands of

years, and an impressive number of modern drugs have been derived from natural

sources, many based on their use in traditional medicine. Preliminary phytochemical

analysis showed that the bark extracts of C. macleodii possess phenolics compounds,

saponins, tannin, coumarines. Phytoconstituents such as saponins, phenolics

compounds and glycosides have been reported to inhibit bacterial growth and to be

protective to the plants producing them against bacterial and fungal infections

(Mather and Gonzalel, 1982).

The extracts of leaves and fruits of Cordia africana were highly effective in

reducing the number of colonies of both bacteria E. coli and Staph. aureus. however

Staph. aureus is more sensitive to the extracts compared to E. coli. The number of

both bacteria colonies is highly reduced.

46

CHAPTER SIX

CONCLUSIONS AND RECOMMENDATIONS

6.1 Conclusions

1. The study proved that the extracts of leaves and fruits of Cordia africana

were more effective on the Gram positive Staph. aureus bacterium than on

the Gram negative bacterium E. coli.

2. The inhibitory effect against the tested bacteria was more effective when

using higher concentrations of the extracts; the effect was decreasing with

increasing dilution.

3. The study indicated that the aqueous extracts of leaves and fruits of Cordia

africana contain antimicrobial activity against the two bacteria tested.

6.2 Recommendation

1. It could be recommended that Cordia africana (gumbil) can be used as

antibacterial activity.

2. Further studies are recommended for the identification the antibacterial

chemical components in the extracts of Cordia africana.

3. The study recommended more attention for benefits from researches made

in the field of medicinal plants and the application and use of their

findings.

47

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