THE ANTIBACTERIAL ACTIVITY OF

THE ROOT AND STEM BARK OF

BRIDELIA FERRUGINEA

By
Luka, Mela Ilu
(UJ/2014/PGMD/0104)

Supervisor; Prof. Yvonne T. Kandakai-Olukemi

INTRODUCTION
The use of plants for medicinal purposes dates
back to antiquity (Ogunyemi, 1998).
WHO has estimated that 80% of the worlds
inhabitant rely chiefly on traditional medicines
(Akerele in Olukemi, 1997).
Majority of plant-derived drugs were
discovered as a result of the correlation
between the use of plant in traditional
medicine and drugs obtained from them
(Olukemi and Kandakai-Olukemi, 2005).
Out of 119 plant-derived drugs, 88(74%) were
discovered as a result of studies to isolate
active substance responsible for the plants use
in traditional medicine (Fansworth et al., 1985).

INTRODUCTION CONTD
Bridelia ferruginea belongs to the
family Euphorbiaceae.
It is usually a gnarled shrub which
sometimes reaches the size of a tree
under suitable conditions.
The tree is 6 - 15 m high, up to 1.5 m
and branching low down. The bark is
dark grey, rough and often scaly
(Rashid et al., 2000).
Its common names are Ola (Igbo), Kizni
or kirni (Hausa), Marehi (Fulani), Ira
lodan (Yoruba)

INTRODUCTION CONTD
The bark extract was reported of having
potential for water treatment (Kolawole and
Olayemi, 2003)
The root of the plant is used as chewing
sticks and the root bark is used for intestinal
and bladder disorder remedies as well as skin
diseases (De-Bruyne et al., 1997).
The bark decoction is used for toothache and
in Ivory Coast for dysentery and diarrhoea or
as a laxative (Gill, 1992).
The bark, leaves and roots are ingredients of
Yoruba (in Nigeria) infusions chiefly
administered to children (Burkil, 1994).

STATEMENT OF THE PROBLEM

The occurrence of multiple antibiotics
resistance developed by microorganisms
against the available synthetic antibiotics
has increased astronomically in the last
decade (Jones and Pfaller, 1998).
The development of this resistance to
drugs calls for active search for more
effective as well as affordable
antimicrobial agents.
This problem has prompted tremendous
effort to explore for more potent
antimicrobial agents, especially of natural
origin to counter this resistance.

STATEMENT OF THE PROBLEM CONTD

Bridelia ferruginea may provide

natural source of antimicrobial that
will provide novel compounds that
may be employed in controlling
some infections globally.

JUSTIFICATION
Literature reports and ethnobotanical
records suggest that Bridelia ferruginea
have tremendous potentials as important
sources for remedy of certain bacterial
infections and of new compounds for
antimicrobial drugs syntheses .
Herbal preparations of Bridelia
ferruginea, though found to be useful in
treating some conditions but the extent
of use is often met with a major setback
from the fact that no scientific evidence is
available for many of such preparations.

JUSTIFICATION CONTD
This study aimed at finding and
establishing a scientific basis for
the use of the plant in search of
effective and affordable cure to
certain bacterial infections.

RESEARCH HYPOTHESIS
The chemical components in the root and
stem bark of Bridelia ferruginea is
responsible for the antimicrobial activity.
There is significant difference between
Minimum Inhibitory Concentration (MIC) and
Minimum Bactericidal Concentration (MBC)
of aqueous and ethanolic extract of the root
of Bridelia ferruginea against the test
organisms.
There is significant difference between MIC
and MBC of aqueous and ethanol extract of
the stem of Bridelia ferruginea against the
test organisms.

AIM OF THE STUDY

The aim of this study is to
determine the antibacterial activity
of aqueous and ethanol extracts of
the root and stem bark of Bridelia
ferruginea.

SPECIFIC OBJECTIVE
To identify some of the phytochemical
properties of the root and stem bark
extracts of Bridelia ferruginea
To determine the Minimum Inhibitory
Concentration (MIC) of the aqueous and
ethanol extract of the root and stem bark of
Bridelia ferruginea against some selected
gram positive and gram negative bacteria.
To determine the Minimum Bactericidal
Concentration (MBC) of the aqueous and
ethanol extract of the root and stem bark of
Bridelia ferruginea against the test
organisms.

MATERIALS AND METHODS

Collection of plant materials;
Fresh root and stem bark of
Bridelia ferruginea will be collected
from the field and identified in the
Herbarium of Federal College of
Forestry, Jos, Plateau State.

Preparation of extracts
Fresh root and bark of Bridelia
ferruginea will be air dried at room
temperature for three to five days
then oven-dried at 50oC for 5 - 7
days to remove the residual
moisture.
The dried root and bark will then be
pulverized using a mechanical
grinder and the powder stored in an
air-tight container until needed for
use.

Preparation of extracts
contd

The root will also be extracted

using the same process. In each of
these case, the solution will be
filtered using Whatmann No 1 filter
paper after allowing it to stand
overnight.

Phytochemical Screening
The extract of the plants will be
analyzed for the presence of
Alkaloids, Saponins, Tannins,
Cardiac Glycoside, Anthraquinones,
Steroid, and flavonoids, according
to standard methods (Ngbede et.
al., 2008) as described below;

Test for Alkaloids

3 grams of extract will be stirred
with ethanol containing 3% tartaric
acid.
The filtrate is then shared into 3
beakers and tested for alkaloids as
follows-Hagars reagent, Mayers
reagent, and Marquins reagent will
be added and in the first, second,
and third beaker respectively.
Precipitations in any of 3 tests will
indicate the presence of alkaloid.

Test for Tannins

In to 10ml of freshly prepared 10% potassium
hydroxide (KOH) in a beaker, 0.5 g of extract
will be added followed by shaking to dissolve.
The appearance of a dirty precipitate
indicates the presence of tannin.

Test for steroids

100mg of extract will be dissolved in 2 ml of
chloroform.
Sulphuric acid is then carefully added to form
a lower layer.
A reddish brown colour at the interface is
indicative of the presence of steroidal ring

Test for Saponins

0.5 g of the plant extract will be
shaked with water in test tube,
frothing which persists on warming
is considered as preliminary
evidence for the presence of
Saponins.
Few drops of olive oil will be
added to the extract and shake
vigorously.
Formation of soluble emulsion in
the extract indicates the presence

Test for Anthraquinones

0.5 g of the extract will be added in
to dry test tube followed by
addition of 5 ml chloroform and the
mixture shaken for 5 min.
The extract is filtered and the
filtrate shaken with an equal
volume of 100% ammonia solution.
A pink violet or red colour in the
ammoniacal layer (lower layer)
indicates the presence of free
anthraquinones.

Test for Cardiac glycoside

100 mg of extract will be dissolved
in 1 ml of glacial acetic acid
containing one drop of ferric
chloride solution.
This is then under layered with 1 ml
of concentrated sulphuric acid.
A brown ring obtained at the
interface indicates the presence of
de-oxysugar characteristics of
cardenolides.

Test for Flavonoids

2 g of the powdered leaves root
and stem bark will be completely
mixed with acetone.
The residue will then be extracted
in warm water after evaporating
the acetone in water bath.
The mixture it then filtered while
still hot followed by cooling the
filtrate before use.
5 ml of 20% sodium hydroxide will
be added to equal volume of the

The Test Organisms

The bacteria to be used for this study
include six clinical isolates; Escherichia
coli, Pseudomonas aeroginosa, Proteus
mirabilis, Staphylococcus aureus, Bacillus
cereus and Stapylocoocus epidermidis.
Escherichia coli NCTC 10418 and
Staphylococcus aureus NCTC 6571 will
serve as a control strains.
This will be obtained from NVRI, Vom, Jos.
The pure cultures of the test organisms
will be prepared in nutrient broth in test
tubes and will be kept in the refrigerator
at 4oC until needed for use.

Preparation of the ethanol and

aqueous of the root and stem
bark the plant

5g each of the stem bark and root

of Bridelia ferruginea will be
weighed and dissolve in 50 ml of
95% ethanol and water to form
100mg/ml concentration.
This forms the stock solution. The
extraction will be done by gentle
and continues agitation of the
mixture for 3hrs.
The mixture will then be filtered.

Culture media to be used

Nutrient agar and nutrient broth
media will be used as culture
media.
Nutrient broth will be used for the
determination of Minimum
Inhibitory Concentration (MIC).
Nutrient agar will be used for the
determination of Minimum
Bactericidal Concentration (MBC).
The media will be prepared and
sterilized as instructed by the

Determination of Minimum
Inhibitory Concentration (MIC)
The tube dilution method described
by Scott, 1999 will be used.
9 tubes labelled 1-9 with each tube
containing 5ml of nutrient broth.
5ml of the crude extract of the root
in the desired concentration will be
introduced to tube 1 and mixed
thoroughly.
5ml of the content in tube 1 will
then be transferred to tube 2 and
subsequently up to tube 8.

Determination of Minimum Inhibitory

Concentration (MIC) Contd
Tube 9 will serve as a control with no
extract.
To all the tubes (1-9), 0.02ml of 24hr
broth cultures of the test organisms
diluted to yield an inoculum equivalent
to 0.5 Mcfarland standard will be added.
The tubes will then be incubated at 37 0C
for 24hr after which they will be
examined for microbial growth.
The MIC of both extract of the root and
stem bark will then be determined.

Determination of Minimum
bactericidal Concentration (MBC)
The MBC will be determined by
culturing the tubes that showed no
growth during MIC determination.
The lowest concentration at which
no growth is observed on the agar
will be noted as the MBC.

Data Analysis
Data will be analysed using SPSS
statistical software version 20.0
Result will be presented using
charts and tables as appropriate.

Selected Reference
Owoseni AA, Ogunnusi T (2006). Antibacterial
Effects of Three Selected Chewing Sticks Extracts
on Lactobacillus sp. Int. J. Trop.Med. 1(3): 103-106.
Shears P (1993). A review article of bacterial
resistance to antimicrobial agents in tropical
countries. Ann. Trop. Paediatr. 13: 219-226.
Jones RN, Pfaller MA (1998). Bacterial resistance: a
worldwide problem. Diagnostic Microbiol. Infectious
Dis., 31: 379-88.
Hart CA, Kariuki S (1998). Antimicrobial resistance
in developing countries. Brit. Med. J., 317: 647-650.
Sofowora EA (1982). Medicinal Plants and
Traditional medicine in Africa.Spectrum Books Ltd.
John Wiley and Sons, Chichester, United Kingdom
pp. 159-189.

Selected Reference
Malcolm SA, Sofowora EA (1969). Antimicrobial
activities of selected Nigerian Folk remedies and
their constituent plants. Lioydia 32: 512-517.
Akubue PI, Mittal GC (1982). Clinical evaluation
of a traditional herbal practice in Nigeria: a
preliminary report. J. Ethnopharmacol. 6: 355359.
Iwu MM (1980). Anti-diabetic properties of
Brideliaferruginea. Plant Med. 39: 247.
Gill LS (1992).Ethnobotanical uses of Plants in
Nigeria.Univesity of Benin Press.
Burkil HM (1994). The Useful Plants of West
Tropical Africa.Vol. 2.The Royal Botanic Garden,
Kew.pp. 636.

Thank
You For
Listenin
g