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Antibacterial activity of Garlic (ALLIUM SATIVUM) on Staphylococcus aureus

and Escherichia coli

Article · October 2017

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 International Journal of Current Science and Studies (IJCSS)
www.ijcss.com

Antibacterial Activity of Garlic (Allium sativum) on

Staphylococcus Aureus and Escherihia Coli
Tijjani, A1, Musa, D.D.2, Aliyu, Y.3

Department of Biological Sciences, Federal University Dutsin-Ma, Katsina Nigeria

Abstract: Studies were carried out to determine the antibacterial activity of Garlic ethanol, and aqueous extracts on Staphylococcus
aureus and Escherichia coli, Media was prepared according to the producing companies instructions and sterilized in autoclave at
121oC under pressure of 15 PSI after incubation at 37oC for 24 hrs, fresh forms of Allium sativum (Garlic) were purchased from
Dutsin-ma market Katsina state. The MICs of the garlic extract was determined using 2ml of different concentrations of the garlic
extract was added to 18 ml of pre-sterilized molten nutrient agar at 40°C. Result obtained revealed that the aqueous and ethanol extracts
of Garlic has antibacterial activity on the test organisms. Hence have the potentials to be used as a template for the development of new
class of antibiotics.

Keywords: Anti-bacterial, Garlic, Ethanol, Aqueous, Allium sativum

1. Introduction
A medicinal plant can be defined as any plant in which one
or more of its parts contains substances that can be used for
therapeutic purposes or which are precursors for the
synthesis of useful drugs (Burkill, 2004). The use of natural
plant products is not new. In fact, it dates back to earliest
history. An early school of thought about the medicinal uses
of plants was called "the doctrine of signatures". It was
believed that all plants existed for human use and that the
proper use was clearly recognizable by the shape of the plant
with bladder-like leaves which were good for ailments of the
bladder-organ (Ward and Hetzel, 1999). Plants produce
many organic substances which have potential value in the
treatment of diseases (Akinyemi et al., 2006).
The most important thing about garlic is that, it has not been
known to contain any poison to animal and man as no cases
have been reported.
Gomaa and Hashish, 2003 reported that Allium sativum
(garlic) extract inhibited the growth of Shigela dysenteriae,
Escherichia coli, Staphylococcus aureus, Salmonella typhi.
Galli et al.(1985) in their studies on the antimicrobial action
of extracts of garlic (AIlium sativum) reported the inhibitory
action of garlic extract on gram-positive and gram-negative
bacteria such as Bacilli and Clostridia, yeasts and moulds.
The aim of this research work was to determine the
antibacterial activity of aqueous and ethanolic garlic extract
(Allium sativum) on Staphylococcus aureus and Escherichia
coli.

Sugar produced during the process of photosynthesis are 2. Methodology

converted by a series of well-defined processes, for example,
phosphorylation, to all the other organic substances found in
plants and in so doing incorporate nitrogen, sulphur to give
proteins and glycerides together with many phenolic
compounds, steriods, terpenes, alkaloids and other chemical
substances. Medicinal properties of plants are normally
dependent on the presence of certain phytochemical bases
such as alkaloids, anthraquinones, cardiac glycosides, tannin
and polyphenols (Gundiza, 1985). Allium sativum (garlic)
belongs to the family Liliaceae and genus Allium. It is a
hardy perennial herb with narrow leaves. It has bulb-like
root consisting of several bulbils (cloves) and the bulb is
milkish in colour. It has a strong smelling, powerful arid
penetrating odour and is used as a condiment in fish, meat
preparations and in various fruit and vegetable preserves
(Dutta, 1998). Allium sativum (garlic) has been extensively
found in India, China, Asia, Southern Europe, North
America and the other Northern part of Nigeria. Garlic
belongs to the genus Allium which is a general name for all
substances which form allicins through enzymatic action.
When fresh garlic is crushed, a characteristic odour is given-
off which is mainly due to the disulphides compound allicin.
Allins are odourless organic disulphides or their oxides and
allicins are formed from allin by the action of enzyme
allinase. Allicins have been reported to have antibiotic
activity and it is considered to be responsible for most of
garlic's pharmacological activities (Stoll and Seebeck, 2006).
Culture media
Media was prepared according to the producing companies
instructions and sterilized in autoclave at 121oC under
pressure of 15 PSI after incubation at 37oC for 24 hrs, used
for culture and diagnosis of bacteria used in this study
(Forbes et al., 2007).
Collection of Plant Material
The fresh forms of Allium sativum (Garlic) were purchased
from Dutsin-ma market Katsina state.
Tested microorganism
The test microorganism (Staphylococcus aureus and E. coli)
that was used in this study was obtained from the
Microbiology laboratory of Federal University Dutsin-ma.
The bacterial was then isolated and first sub-cultured in a
nutrient agar and incubated at 37°C for 24hours.
Extraction from fresh garlic
Fifty grams (50g) of garlic was crushed and socked in 100ml
distilled water in a sterile sample bottle and kept at room
temperature for 4days. The socked garlic was then filtered
using filter paper (Whatman no.1). Different concentration
of the garlic extract was made by diluting the crude garlic
with distilled water such as 500mg/ml, 400mg/ml,
300mg/ml, 200mg/ml, 100mg/ml and 50mg/ml. The Ethanol

Volume 1 Issue 1, October 2017

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Paper ID: 14101703 47
 International Journal of Current Science and Studies (IJCSS)
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extract was prepared following same procedure with the Table 2: phytochemical screening of ethanol extract of
exception of solvent which was 95% ethanol instead of Allium sativum
sterilized distilled water. Phytochemical compd. Indication
Tannins -
Garlic extracts activity test well diffusion method Saponins +
screening Cardiac glycosides +
Alkaloids +
Screening of the anti-bacterial activity was performed by Flavonoids -
well diffusion technique (Saeed et al., 2005). The Mueller Key= + present; - absent
Hinton agar plates were seeded with 0.1ml of the
standardized inoculums of bacteria. The inoculums were Table 3: phytochemical screening of water extract of Allium
spread evenly over plate with sterile glass spreader. The sativum
seeded plates were allowed to dry in the incubator at 37oC Phytochemical compd. Indication
for 20 minutes. A standard crack border of 6 mm diameter Tannins +
Saponins -
was used to cut uniform wells on the surface of the plates,
Cardiac glycosides +
and 0.1ml of each concentration was introduced in the well.
Alkaloids +
The inoculated plates were incubated at 37oC for 24hrs and Flavonoids -
zone of inhibition diameter was measured to using nearest Key= + present; - absent
millimeter (mm).
Table 4: Effect of different concentration of aqueous garlic
Determination of the minimum inhibitory concentrations extract on Staphylococcus aureus and Escherichia coli
(MIC) Zone of Inhibition (mm)
CONC (mg/ml) S. aureus E. coli
The MICs of the garlic extract was determined using the 500 0 0
method described by Akinpelu and Kolawole (2004). 2ml of 400 2 4
different concentrations of the garlic extract was added to 18 300 7 10
ml of pre-sterilized molten nutrient agar at 40°C. The 200 13 12
medium was then poured into Petri dishes and allowed to 100 18 15
set. The surfaces of the media were allowed to dry before 50 21 20
streaking with 24hours old bacterial isolates. The plates
were later incubated in an incubator at 37°C for up to Table 5: Effect of different concentration of ethanol garlic
24hours after which they were examined for the presence or extract on Staphylococcus aureus and Escherichia coli
absence of growth. The MIC was taken as the lowest Zone of Inhibition (mm)
concentration that will prevent the growth of the test CONC (mg/ml) S. aureus E. coli
bacterial isolates. 500 0 0
400 12 5
Determination of minimum bactericidal concentrations 300 17 10
(MBC) 200 20 12
100 25 17
50 30 25
The MBC of the extract was determined using Olorundare et
al., (1992) with little modification. Samples were taken from
plates with no visible growth in the MIC assay and sub- Table 6: MIC result for aqueous extraction
CONC (mg/ml) S. aureus E. coli
cultured onto freshly prepared nutrient agar plates and later
500 - -
incubated at 37°C for 24 hours.
400 - -
300 - -
The MBC was taken as the concentration of the extract that 200 + +
did not show any growth on a new set of agar plates. 100 + +
50 + +
3. Result Key: + no inhibition, - there is inhibition
The result for MIC is at 300mg/ml.
Table 1: Characteristics of the garlic extract used
Ethanol extracts of Aqueous extract of Table 7: MIC result for ethanol extraction
garlic garlic CONC (mg/ml) S. aureus E. coli
Weight used 50g 50g 500 - -
Color Brownish Brownish 400 - -
Smell True garlic odor True odor of garlic 300 - -
Volume of 200 - -
100ml (90%) 100ml
solvent 100 + +
50 + +
Key: + no inhibition, - there is inhibition

The result for MIC is at 200mg/ml. The result for MBC is

200mg/ml was observed in ethanol garlic extract while the
MBC of 300mg/ml was observed in aqueous garlic extract.

Volume 1 Issue 1, October 2017

Licensed Under Creative Commons Attribution CC BY
Paper ID: 14101703 48
 International Journal of Current Science and Studies (IJCSS)
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4. Discussion
The result obtained from this study showed that garlic
possessed anti-bacterial effect against Gram positive
(Staphylococcus aureus) and Gram negative (Escherichia
coli) bacteria just as it was described by Louis Pasteur that
Onion and Garlic juices (extract) possess antibacterial
activities against both Gram positive and Gram negative
bacteria (Whitemore and Naidu, 2000).
[11] Saeed, S., and Tariq, P., (2005). Anti-bacterial activities
of Mentha piperita, Pisum sativum and Momordica
charantia, Pak. Journal of Botany., 37:997-213.
[12] Ward, J.A., and Hetzel, H.R., (1999). Biology, Today
and Tomorrow. 2nd Edition, West Publishing Company,
USA. 487 – 504.

The sensitivity of the bacteria was gradually increased with

the increased in the garlic extract concentration. According
to Cheesbrough (1984) an antimicrobial agent which the
diameter of the zone of inhibition is above 3mm, the
organism is said to be sensitive but if it is 2mm or less than
that, the organism is said to have resistant against the
particular agent.

Therefore, considering the statement of Cheesbrough, and

comparing the result obtained in table 1, 2, 3 and 4 above
showed that the ethanol and water extract of garlic (Allium
sativum) have antimicrobial effect of Staphylococcus aureus
and Escherichia coli. This activity is not unconnected with
the presence of the phytochemicals identified.

The result above showed that ethanol extract have higher

antimicrobial activities on both Staphylococcus aureus and
Escherichia coli than water garlic extract due to the diameter
of their zone of inhibition.

References
[1] Akinyemi, K.O., Oluwa, O.K. and Omomigbehin., E.O.
(2006). Antimicrobial Activity of Crude Extracts of
three medicinal plants used in South –west Nigeria Folk
Medicine on some Food Borne Bacterial Pathogens.
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[2] Akinpelu, D.A., Kolawole, D.O., (2004). Phytochemical
and Antimicrobial Activity of leaf extract of Piliostigma
thonningii (Shum.). Sci. Focus 7:64-70.
[3] Burkill, H.M., (2004). The Useful plants of West
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Gardens, Kew. 612.
[4] Dutta A.C. (1998). Botany for Degree Students. Oxford
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[5] Forbes, B.A., Sahm, D.F.,Weissfeld, A.S., (2007).
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[6] Gundiza, M., (1985). Phytochemical Screening of
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[7] Galli, A., Frazetti, L., and Briguglio, D., (1985).
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[8] Gomaa, N.F., and Hashish, M.H., (2003). Journal Egypt
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[9] Olorundare, E.E., Emudianugbe, T.S., Khaar, G.S,
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[10] Stoll, A., and Seebeck, E., (2006). Chemical
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Wiley Online Library, Pg 11 – 35.
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