Int'l Conf.

on Advances in Environment, Agriculture & Medical Sciences (ICAEAM’14) November 16-17, 2014 Kuala 
Lumpur (Malaysia) 

Antibacterial Effect of Garlic (Allium

Sativum) On Staphyloccus Aureus: An In
Vitro Study
Mohamed A. Eltaweel
http://dx.doi.org/10.17758/IAAST.A1114040 47 
Therefore such plants should be investigated
to understand Abstract—The methanol and aqueous suspensions of the dried
their properties, safety and efficacy and for a
search of new Allium sativum (Liliaceae) bulbs extract was screened for its anti-
potent antimicrobial compounds and fractions [11].
microbial activity using the agar-well diffusion method. It is tested against Gram-positive bacteria (Staphylococcus
aureus). The suspensions were tested at concentrations of 1, 10, 100 and 1000 μg/ml . All suspensions showed an
inhibitory effect against tested bacteria. The highest zone of inhibition was estimated with the highest concentration of
aqueous suspension (48 mm) followed by the highest concentration of the methanolic suspensions (1000 μg/ml)
which reached to 34 mm. The other concentrations either methanolic or
The problem of microbial resistance is growing and the outlook for the use of antimicrobial drugs in the
future is still uncertain [12]. In general, bacteria have the genetic ability to transmit and acquire resistance
to drugs [13]. The success story of chemotherapy lies in the continuous search for new drugs to counter
the challenge posed by resistant strains of microorganisms[14]. aqueous showed various inhibitory effect
on the tested bacteria.
II. M
ATERIAL
Keywords—Antimicrobial activity, traditional herbal, Allium sativum, Staphylococcus aureus

I. I
NTRODUCTION Many infectious diseases have been known to be treated with herbal remedies
throughout the history of mankind. Natural products, either as pure compounds or as standardized plant
extracts, provide unlimited opportunities for new drug leads because of the unmatched availability of
chemical diversity [1]. Garlic (Allium sativum) has traditional dietary and medicinal applications as an
anti-infective agent [2], [3].
In vitro evidence of the antimicrobial activity of fresh and freeze-dried garlic extracts against many
bacteria [4], [5], fungi [6], and viruses [7] supports these applications. Although hundreds of plant species
have been tested for antimicrobial properties, the vast majority of have not been adequately evaluated [8].
Recently, medicinal plants have become the focus of intense study regarding their conservation and
potential pharmacological effects. Indeed, the search for new pharmacologically active agents, through
the screening of natural sources such as microbial fermentations and plant extracts, has led to the
discovery of many clinically useful drugs that now play major roles in the treatment of human diseases [9],
[10].
About 80 % of individuals from developed countries use traditional medicine, which has compound
derived from medicinal plants.
Mohamed A. Eltaweel, Faculty of Science, University of Misurata, Libya. e-mail: Eltaweel1963@yahoo.com
A
ND
M
ETHODS
A. Preparations of plant materials Garlic (Allium sativum) used in the present study was purchased from
the local market. Bulbs were peeled and washed from the foreign particles. A Fifty grams of the bulbs
were squeezed. The squeezed sample was sucked at methanol for 8hs with 10 minutes interval shaking.
The extraction was filtered using muslin cloth and then Whatman no. 1 filter paper. The filtrate was
evaporated at 45oC to dryness and the dried substance was kept in sterile bottle under refrigerated
condition until use.
The dry weight of the extract was obtained by allowing the solvent to evaporate and was used to
determine concentration in mg/ml. methodology based on [15]. Four concentrations of each extract (1, 10,
100, and 1000 mg/ml) were prepared by resuspending the dried extract in methanol separately. Other
same concentrations were resuspended in sterile distilled water (aqueous).
B. Tested microorganisms The test microorganisms (Staphylococcus aureus ) used in this study was
obtained from the Misurata Central Hospital Laboratory. The bacterial isolate was first subcultured in a
nutrient broth (Oxoid) and incubated at 37°C for 18 h .
C. Antimicrobial activity The antibacterial activity of the crude extracts was determined in accordance with
the agar-well diffusion method described by [16]. Cup-plate method was used to test the antibacterial
activities of the aqueous and methanolic Allium sativum against the test bacteria isolates.
Eighteen hour broth cultures were diluted appropriately using McFarland scale (0.5 McFarland which is
about106 cfu/ml). The molten sterile nutrient agar (20 ml) was poured into sterile petri dish and allowed to
set.
The sterile nutrient agar plates were flooded with 0.2 ml of the standardized inoculum. A sterile cork
borer (No. 6) was
Int'l Conf. on Advances in Environment, Agriculture & Medical Sciences (ICAEAM’14) November 16-17, 2014 Kuala 
Lumpur (Malaysia) 
http://dx.doi.org/10.17758/IAAST.A1114040 48 used to bore equidistant cups into the agar plate.
One drop of
The methanolic extract showed lower
action compared to the molten agar was used to seal the bottom of the bored hole,
the aqueous extract (resuspension) as
antimicrobial agents. so that the extract will not sip beneath the agar. Approximately
This may be due to little diffusion properties
of the extract in 50 μl of the crude extract at different concentrations (1 mgml1,
the agar or because fresh plants contain
active substances 10 mgml1, 100 mgml1 and 1000 mgml1 ) were introduced into the
which may be affected or attributed by the
used solvent. wells.
Reference [18] reported that 100 mgml-' of
the garlic A control was prepared by putting 50 μl of freshly prepared
aqueous extract shows an inhibition zone of
19.3mm on sterile distilled water in one of bored hole at the plates
Staphylococcus aureus. also reported that
garlic methanol extract containing aqueous suspensions where 50 μl of methanol use at
at concentration of 100mg/ml shows an
inhibition zone of 11mm the plates containing methanolic suspensions. One hour pre-
on the same bacteria. diffusion time was
allowed, after which the plates were
However the inhibitory zone of the same
concentration at this incubated at 37°C for 18 h. The zones of inhibition were then
study showed was higher effect (27 mm)
comparing to the measured in millimeter. The above method was carried out in
mentioned study. This is may be due to the
species difference triplicates and the mean of the triplicate results were taken.
or the garlic difference in different biologic condition. [19]
D. Preparation of Inoculum About 18 hour broth culture of the test bacteria isolate was suspended into
sterile nutrient broth. It was standardized according to National Committee for Clinical Laboratory
Standards [17] by gradually adding normal saline to compare their turbidity to McFarland standard of 0.5
which is approximately 1.0 × 106 cfu/ml.
reported that S. aureus (NSA1) was most susceptible to the active principles present in garlic, which had
a zone of growth inhibition diameter of 28 mm in water, 23, 24 and 32 mm in chloroform, ethanol and
metronidazolem respectively.
Such results were similar from data reported by Vuddhakul [20] who observed that garlic extracts
inhibited the growth of Staphylococcus aureus. In contrast [21]. Reported that crude extracts of garlic did
not exhibit any in vitro inhibition on the
III. RESULTS AND DISCUSSION
growth of test organisms including Staphylococcus spp.
The large sizes of zones of growth inhibition
produced by The resuspension extract aqueous, and methanolic of
the garlic extracts against the tested bacteria
indicated the Allium sativum and (bulbs) was investigated for their
potency of the active principles in garlic.
antimicrobial activity against Staphylococcus aureus. The
The study focused on the garlic
antibacterial activity on S. ―hole plate‖ diffusion method was used in testing various
aureus has shown that dilute solutions of
garlic can concentrations of this extract.
completely inhibit the growth of S. aureus at the
concentration The results depicted in Table 1 indicate that the high
of more than 7.50 mg/ml. This could be due
to the action of concentrations of Allium sativum aqueous extract (1000
biological active ingredient of allicin which
exhibits its mgml') used in this study had the highest inhibitory effects
antimicrobial activity mainly by immediate
and total inhibition (48mm) against the tested microorganisms. However, this
of RNA synthesis, although DNA and protein
syntheses are extract showed inhibition action of 19 mm even at minimal
also partially inhibited, suggesting that RNA
is the primary concentration (1 mg/ml) used in this study. The other
target of allicin action [22]. concentrations of
the aqueous phase (10 and 100 mgml-') gave
Using the same protocol garlic has a
bactericidal effect at an inhibition zones of 23 and 27mm, respectively.
the lower concentration of 30.00 mg/ml for
clinical isolate of Methanolic extract of Allium sativum at all concentrations
S. aureus. However, this concentration level
may vary as shows an inhibitory effect against the Staphylococcus aureus.
different authors have stated; for instance
160 mg/ml was The highest inhibition zone obtained was 34 mm with the
observed by [23]. concentration of 1000
mgml-'. Concentrations of 1, 10 and 100 mg/ml-' showed an inhibition zones of 16, 17 and 21 mm,
IV. C
ONCLUSION
respectively.
TABLE I A
NTIBACTERIAL
A
CTIVITIES
P
ROFILE
O
F
A
LLIUM
S
ATIVUM
E
XTRACT RESUSPENSIONED IN METHANOL AND STERILE
DISTILLED WATER AGAINST
It is concluded from this study that Allium sativum extract has antimicrobial activity against
Staphylococcus aureus. It is expected that using natural products as therapeutic agents will
STAPHYLOCOCCUS AUREUS BACTERIA.
probably not elicit resistance in microorganisms. It is essential Zone of
inhibition (mm)
Extract suspension
(1 μg/ml)
(10 μg/ml) (100 μg/ml)
(1000 μg/ml)
that research should continue to isolate and purify the active components of this natural herb and use in
experimental Methanolic Aqueous 16± 0.3 17 ± 0.5 21 ± 0.6 19 ± 0.9 23 ± 0.5 27 ± 0.3 34 ± 1.0 48 ± 0.6
animals.
This study showed that aqueous extract phase of Allium
REFERENCES
sativum gave better results compared to the metanolic phase of
[1] P. J. Parekhm and S. V. "Chandam In vitro
Antimicrobial Activity and the same extract at this study at all concentration tested . The
Phytochemical Analysis of Some Indian
Medicinal Plants", Turk J Biol., broad antimicrobial action of the aqueous extract of the Allium sativum
used in the study could be ascribed to the water soluble components which are naturally occurring in the
plant materials.
31: 53-58, 2007. [2] L. D. Lawson, "Garlic: a review of its medicinal effects and indicated active compounds", p.
176–209. In L. D. Lawson and R. Bauer (ed.), 1998, Phytomedicines of Europe: their chemistry and biological activity.
ACS Symposium Series, no. 691. American Chemical Society, Washington, D.C.
Int'l Conf. on Advances in Environment, Agriculture & Medical Sciences (ICAEAM’14) November 16-17, 2014 Kuala 
Lumpur (Malaysia) 
[3] H. D Reuter, H. P. Koch, and L. D. Lawson. " Therapeutic effects and applications of garlic and its preparations",
In H. P. Koch and L. D. Lawson (eds.) 1996, Garlic. The science and therapeutic application of Allium sativum L. and
related species–2nd Edn., pp: 135-212.Williams & Wilkins, Baltimore, Md. [4] C. J. Cavallito, and J. H. Bailey "Allicin,
the antibacterial principle of Allium sativum. I. Isolation, physical properties, and antibacterial action", J. Am. Chem.,
Soc. 66:1950, 1944. [5] L. P. Rees, S. F. Minney, N. T. Plummer, J. H. Slater, and D. A. Skyrme. "A quantitative
assessment of the antimicrobial activity of garlic" (Allium sativum). World. J. Microbiol. Biotechnol., 9:303–307, 1993.
[6] M. Adetumbi,, G. T. Javor, and B. H. S. Lau. "Allium sativum (Garlic) inhibits lipid synthesis by Candida albicans",
Antimicrob. Agents Chemother, 30:499–501, 1986. [7] N. D. Weber,, D. O. Anderson, J. A. North, B. K. Murray, L. D.
Lawson, and B. G. Hughes, "In vitro virucidal effects of Allium sativum (garlic) extract and compounds", Planta Med.,
58:417–423, 1992. [8] M. F. Balandrin, J. A. Klocke, E. S. Wurtele and W. H. Bollinger, ''Natural plant chemicals:
Sources of Industrial and Medicinal materials''. Science, 228: 1154-1160, 1985. [9] S. Yue-Zhongm "Recent natural
products based drug development. A pharmaceutical industry perspective", J. Nat. Prod., 61: 1053 – 1071, 1988. [10]
S. G. Leitao,, O. Castro,, E. M. Fonseea,, L. S. Juliao,, E. S. Tavares, and R. R. Leo, R. C. Vieira,,.D. R. Oliveira, G.
G. Leitao,, V. Martino,, V. Sulsen,, Y. A. Barbosa,, D.P. Pinheiro,, P. E. Silva,, D. F Teixeira, and M. C. Lourenco,
"Screening of general and south American plantsextracts for antimycobacterial activity by the Alamar Blue test", Rev.
Bras. Farmacogn., 16: 6 – 11, 2006. [11] J. N. Ellof, "Which extractant should be used for the screening and isolation
of antimicrobial components from plants?," Journal of Ethnopharmacology, 60: 1-8, 1998. [12] G. F. N. Gislene, L.
Juliana, C. F. Paulo and L. S. Giuliana "Antimicrobial activity of plant extracts and photochemical on antibiotic
resistant", Brazilian Journal of Microbiology, 31:247-256, 2000. [13] M. L. Cohen, "Epidemiology of Drug Resistance:
Implications for a
Post- antimicrobial" Era. Sci., 257: 1050-1055, 1992. [14] Z. Y. El Astal, A. E. R. A. Ashour, and A. A. M. Kerrit, "
Antimicrobial activity of some medical plant extracts in Palestine", Pakista Journal of Medical Science, 21 (2) 187-93,
2005. [15] J. E. C. Betoni, R. P. P. Mantovani, L.C. Di Stasi, A. Fernandes Junior, "Synergism between plant extract
and antimicrobial drugs used on Staphylococcus aureus diseases", Mem. Inst. Oswaldo Cruz., 101: 387- 390, 2006.
[16] O. Irobi, M. Moo-Young., W. Anderson, S. Daramola "Antimicrobial activity of the bark of Bridelia ferruginea
(Euphorbiaceae)", Intern. J. Pharmacog, vol. 34 pp. 87-90, 1994. [17] National Committee for Clinical Laboratory
Standard, Performance standard for antimicrobial disc susceptibility testing, Twelfth [International Supplement;
Approved standard M100-S12, National Committee for Clinical Laboratory Standards, Wayne, Pa, 2002. [18] G. Iram,
S. Mariam,, S. Halima,, M. A. Shahbaz, , Q. S. Zahoor and A. M. Athar "Inhibitory effect of Allium sativum and
Zingiber officinale extracts on clinically important drug resistant pathogenic bacteria", Annals of Clinical Microbiology
and Antimicrobials, 8-11. 2012. [19] M. A. EL-mahmood, "Efficacy of crude extracts of garlic (Allium sativum Linn.)
against nosocomial Escherichia coli, Staphylococcus aureus, Streptococcus pneumoniea and Pseudomonas
aeruginosa", Journal of Medicinal Plants Research, Vol. 3(4) 179-185, 2009. [20] V. Vuddhakul, P. Bhooponga, F.
Hayeebilana, and S. Subhadhirasakulb "Inhibitory activity of Thai condiments on pandemic strain of Vibrio
parahaemolyticus", Food Microbiol., 24: 413-418, 2007. [21] R. A Onyeagba, O. C. Ugbogu, C.U. Okeke, and O.
Iroakasi "Studies on the antimicrobial effects of garlic (Allium sativum Linn), ginger (Zingiber officinale Roscoe) and
lime (Citrus aurantifolia Linn)", Afr. J. Biotechnol., 3(10): 552-554, 2004. [22] R. Feldberg, S. Chang, A. Kotik, M.
Nadler, Z. Neuwirth, D. Sundstrom and N. Thompson "In vitro mechanism of inhibition of bacterial growth by allicin.
Antimicrob", Agents Chemother, 32: 1763- 1768, 1988. [23] G. Sivam, J. Lampe, B. Ulness, S. Swanzy and J Potter
"Helicobacter pylori-in vitro susceptibility to garlic (Allium sativum) extract", Nutr. Cancer, 27: 118-121, 1997.
http://dx.doi.org/10.17758/IAAST.A1114040 49