Mycopath (2009) 7(1): 51-57
Antifungal activity of some medicinal plants
used in Jeddah, Saudi Arabia
Fardos M. Bokhari
Faculty of Sciences, Biology Department, King Abdel Aziz University,
P. O. Box 12161, Jeddah 21473, Saudi Arabia
* Corresponding author’s e-mail: email@example.com
Development of more effective and less toxic antifungal agents is required for the treatment of
dermatophytosis. Plants and their extraction preparations have been used as medicines against infectious
diseases. In this research, the lemon grass [Cymbopogon citrates DC.) Stapf.], lantana (Lantana camara
L.), nerium (Nerium oleander L.), basil (Ocimum basilicum L.) and olive leaves (Olea europaea L.) were
extracted with either water or different organic solvent to investigate their antifungal activities in vitro. The
methanol extract of lemon grass, lanta and nerium followed by their ethyl acetate extracts showed the
highest activities against Trichophyton rubrum. These inhibited the growth of T. rubrum by 85-90 and 80-
85%, respectively at a concentration of 100 µg ml-1, while aqueous extracts inhibited the growth of this
fungus at the same concentration by 32-77%. The activity of the methanolic extracts of the 5 selected
plants was determined against different pathogenic fungi including Microsporum canis, M. gypseum, and T.
mentagrophytes. Extracts of lemon grass were the most effective followed by lantana. . Nerium and basil
showed moderate activities. The lowest activity was recorded for olive extract. The five dermatophytes
differed with regard to their susceptibility to plant extracts. Trichophyton rubrum was the most susceptible
dermatophyte, followed by Microsporum canis, M. gypseum, and T. mentagrophytes, respectively. The
MICs of these most active plants ranged from 25 to 125 µg ml−1. In conclusion, ethanolic extracts of some
medicinal plants can be used to treat infections with pathogenic fungi.
Key words: Antifungal activities, dermatophytes, MIC, Microsporum, Trichophyton.
Skin, hair, nail, and subcutaneous tissues in
human and animal are subjected to infection by
several organisms, mainly fungi named
dermatophytes and cause dermatophytoses
(Valeria et al., 1996; Amer et al., 2006).
Dermatophytoses are one of the most frequent skin
diseases of human, pets and livestock (Tsang et
al., 1996). The disease is widely distributed all
over the world with various degrees and more
common in men than in women. There are three
genera of mould that cause dermatophytosis.
These are Epidermophyton, Trichophyton and
Microsporum. Contagiousness among animal
communities, high cost of treatment, difficulty of
control and the public health consequences explain
their great importance (Chermette et al., 2008). A
wide variety of dermatophytes have been isolated
from animals, but a few zoophilic species are
responsible for the majority of the cases, viz.
Microsporum canis, Trichophyton
mentagrophytes, Trichophyton equinum and
Trichophyton verrucosum, as also the geophilic
species Microsporum gypseum (Hasegawa, 2000;
Mahmoudabadi and Zarrin, 2008). According to
the host and the fungal species involved, the
typical aspect of dermatophytic lesions may be
modified. A few antifungal agents are available
and licensed for use in veterinary practice or
human being treatment. The use of systemic drugs
is limited to treat man or animal due to their high
toxicity and problems of residues in products
intended for human consumption (Araujo et al.,
2009). Different treatments have been
recommended to control dermatophtes. In general,
pharmacological treatment option include
antifungal agents [Aly, 1997; Agwa et al., 2000],
but recently the use of some natural plant products
has been emerged to inhibit the causative
organisms. The antimicrobial and antitoxin
properties of some plants, herbs, and their
components have been documented since the late
19th century (Saadabi, 2006). These natural plants
involve garlic, lemon grass, datura, acacia, a
triplex, ginger, black seed, neem, basil, eucalyptus,
alfalfa and basil (Omarand Abd-El-Halim, 1992;
Aly et al., 2000; Aly and Bafiel, 2008). They are
safe to human and the ecosystem than the chemical
antifungal compounds, and can easily be used by
the public who used them for thousands of years to
enhance flavor and aroma of foods as well as its
economic value (Shelef et al., 1980; Shelef, 1983).
52 Fardos M. Bokhari
Early cultures also recognized the value of these
plant materials in medicine . Plant extract has
been used traditionally to treat a number of
infectious diseases including those caused by
bacteria, fungi, protozoa and viruses (Soylu et al.,
2005; Yoshida et al., 2005; Nejad and Deokule,
2009). A number of reports are available in vitro
and in vivo efficacy of plant extract against plant
and human pathogens causing fungal infections
(Natarajan et al., 2003). The activity of plant
extract against dermatophytosis i.e. the superficial
infections of skin or keratinised tissue of man and
animals can be very well visualized from the
reports of Venugopal and Venugopal (1995).
They reported the activity of plant extracts against
88 clinical isolates of dermatophytes which
includes Microsporum cannis, M, audouinii
Trichophyton rubrum T mentagraphytes, T
violaccum, Tsimii, T verrucosum T erinacci and
Epidermophytn floccosum by agar dilution
technique. While Vlietinck et al. (1995) reported
clinical findings of Rwandese medicinal plants
(267 plant extracts) used by traditional healers to
treat microbial infections and found 60% of these
extracts were active against dermatophytes. All
the above reports and many others have utilized
plant extract, juice or oil for the in vitro or in vivo
evaluation of the infections caused by various
species of dermatophytes viz. Trichophyton,
Microsporum, Epidermophyton and yeast like
fungi of genera Canddia, Cryptococcus,
Rhodotorula and Torulopsis trichosporon. Up to
now more than 200 different biologically active
substances have been isolated from plant extract,
among them organosulphur compounds such as
allicin, azoenes and diallyltrisulfide. Eugenol,
phenolic compound, the most important
biologically active compound found in many plant
extract (Kähkönen et al., 1999; Aly and Bafiel,
The present study was designed to evaluate
the in vitro antidermatophyte activity of some
plant extracts. The antifungal activities of water
and organic plant extract are compared. The
percentage of inhibition and MIC are also
Materials and Methods
The fungi used were obtained from the
culture collection of Dr. R. Bonally, Laboratoire
de Biochemie Microbienne, Fac. De Pharmacie,
Nancy, France. Microsporium ferruginum,
Trichophyton mentagrophytes and
Epidermatophyton sp. were isolated and identified
from contaminated dust samples collected from
different hospitals in Garbia, Egypt (Amer et al.,
2006). All fungi were stored on sabouraud
dextrose agar (Oxoid) slants in the refrigerator at 4
°C prior to use.
Medicinal pant materials
Samples of five medicinal plants, i.e. basil
leaves (Ocinum bacilicum), lantana leaves and
flowers (Lantana camara), lemon grass stalk and
leaves (Cymbopogon citratus), nerium leaves
(Nerium oleander) and olive leaves (Olea
europaea) were collected during October 2007
from different districts of Jeddah city, Saudi
Arabia and identified by Botany department,
Faculty of Sciences, Tanta Uni., Egypt. The plants
were brought to the laboratory and thoroughly
washed in running tap water to remove debris and
dust particles and then rinsed in distilled water.
Preparation of aqueous and organic medicinal
For aqueous and organic extraction, 10
grams of each sun-dried medicinal plant material,
were cut into small pieces and then macerated by
blender 1–2 mm separately and the powder
produced was blended with 100 ml of either
distilled water (cold or hot) or organic solvent
(ethyl alcohol, methanol, n-butanol, ethyl acetate
or chloroform), (1:10 w/v). Then, they were
extracted under cold conditions for 24 h. The
resultant extract was filtered through a glass wool
filter and then rinsed with a small quantity (about
30 ml) of 96% ethyl alcohol. The extracts
solutions were evaporated under reduced pressure
at 40 °C. Subsequently, the extracts were diluted
by distilled water and stored in the deep freezer at
-10 °C and later lyophilized in a freeze dryer.
Antimicrobial activity of the above
mentioned extracts was determined, using the agar
well diffusion assay method as described by
Holder and Boyce (1994). Dimethyl sulfoxide
DMSO was used as a negative control and
Griseofulvin was used as a positive control. The
plates were done in triplicates and were incubated
at 37 °C. The antimicrobial activity was taken on
the basis of diameter of zone of inhibition, which
was measured after 7 days of incubation and the
mean of three readings is presented. The presence
of inhibition of the treated fungus was calculated
using Griseofulvin as standard (100% inhibition).
The plant extract and the standard antifungal
Mycopath (2009) 7(1): 51-57
Antifungal activity of some medicinal plants 53
agents were dissolved in DMSO, 100%
biologically inert substances.
Determination of minimal inhibitory concentra-
tion of plant extract on fungal growth
The MIC was determined by the methods
described by Chand et al. (1994) and modified by
Aly (1997). Each well of a 96 well ELISA tray
was filled with 175µl of an exponentially growing
culture (106~107CFU ml-1). To each well, 20 µl
solution of each concentration of the test
substance, or the appropriate solvent as control,
was added. The ELISA trays were incubated for
40 minutes before 5 µl of a 0.2% w/v solution of
Fluorescein diacetate (FDA) in acetone was added.
Incubation was continued for 90 minutes more and
the resulting green color from the hydrolysis of
FDA was measured at 490 nm (referenced to 630
nm) and blanked against control wells containing
microbial cultures only, using an MR7000
automatic ELISA tray reader. The agar plates
were incubated overnight (37 °C) and CFU was
counted using a colony counter. The MIC
corresponded to the minimum concentration of the
compound that caused 99% cell inhibition with
respect to the CFU's in a control which contained
microbial cultures and sterile distilled water or
solvent replacing the test compound.
Each experiment has three replicates and
three determinations were conducted. Means of
variable and standard deviation were recorded.
Results and Discussion
Many investigations were carried out to
discover plant products that inhibit the fungi like
Trichophyton rubrum and Microsporum canis.
These two species cause common infections in
humans which are difficult to control effectively,
and the pharmaceutical arsenal currently available
against them is rather limited (Evans and White,
1967; Levine, 1982; Gupta et al., 1991; Jansen et
al., 1991). Hence, plant products that inhibit their
growth without harming the host represent
potential therapeutic agent. As stated earlier, five
different plants belonging to different families
(Table 1) used rationally by Saudi Arabia people
were collected from Jeddah, and extracted with
water or organic solvents and their antifungal
activities were detected against T. rubrum which is
considered one of the fungi usually causes disease
in keratinized epithelial structures such as hairs
and nails and can invade the dermis, particularly in
immunocompromised patients (Maoz and
Neeman, 1998). The antifungal activities of the
plant extracts obtained using different organic
solvents were compared with that of Griseofulvin
and the % of inhibition was calculated (Table 2).
Extracts were obtained through the extracting
action of the appropriate solvent on a dry plant and
the active compounds are thus contained in the
solvent used. Each type of extract is defined by
the way it is prepared and the nature of the solvent.
The extraction process is always studied to respect
the integrity of the active molecules. In this
experiment, Methanol extract of lemon grass was
the best to suppress the growth of T. rubrum (90%
inhibition), followed by lantana and nerium
methanolic extract (85-88% inhibition). The
methanol extract of basil as well as olive inhibited
T. rubrum growth by 73-75%. Extraction of basil
and olive with chloroform was found to be the best
(77-80% inhibition) compared to the other extracts
due to the presence of some essential oil which
could be extracted with chloroform. Extraction of
lemmon grass, lantana or nerium with either ethyl
acetate extract, n-butanol or diethyl ether was less
active against T. Rubrum compared to their ethanol
extract. Aqueous extract of cold or hot water of all
examined plants showed the lowest activity against
T. Rubrum compared to different organic solvents
The activity of methanol extract of the five
selected plants against different dermatophytes
were summarized in Table 3. It was found that
lemon grass extract showed maximum antifungal
activity against T. mentagrophytes, followed by T.
verrucosum, M. canis and E. floccosum (Table 3).
Moderate activity was recorded against different
dermatophyted by using lantana. Less activities
were recorded for nerium, basil and olive. Plant
derived compounds are of interest in this context
because they comprise safer or more effective
substitutes for synthetically produced
antimicrobial agents (Dupuis et al., 1972). The
plant extracts used in folkloric medicine in
Palestine, Saudi Arabia (Abdulmoniem, M. A. and
Saadab, 2006; Aly and Bafiel, 2008), Egypt (El-
Fadaly et al., 1999), Mexico (Navarro et al., 1996)
and India (Jain et al., 2004) were investigated for
their antifungal activity and their use to treat
pathogenic fungi. Lemon and lantana extracts
showed excellent antidermatophytic properties
compared to other plant extract which may be due
to free and bound flavonoid fractions, showed the
greatest fungicidal properties. The maximum zone
of inhibition was recorded in the presence of free
flavonoid fraction of the plant extract against T.
rubrum and T. terrestre, which were the most
susceptible fungus for all the extracts tested (Jain
Mycopath (2009) 7(1): 51-57
54 Fardos M. Bokhari
et al., 2004). Lemmon grass extract has shown
itself to be among the most significant of these
newly uncovered natural, nontoxic therapies, and
has proven itself to be one of the most important
antimicrobial agent successfully used for
treatments of all kinds of infections arising from
fungi, virus, bacteria, parasites, and other
microscopic invaders (Mohamed et al., 2006).
The methanol extracts of lantana (leaves and
flowers) showed antifungal activity (20 mm)
against M. gypseum, T. mentagrophytes, M. canis,
and T. gypseum. On the contrary, olive extract
showed the lowest activity against all tested
dematophytes (8-10 mm). More activities by olive
leaves were recorded against plant pathogenic
fungi including, Alternaria solani, Botrytis cinerea
and Fusarium culmorum (Winkelhousen et al.,
2005). They added that the activity was attributed
to the presence of phenolic compounds which can
be hold a good promise as a natural fungicide
against common pathogens of crops. Nwachukwu
and Umechuruba (2006) found that Leaf extracts
of neem, basil, bitter leaf and paw-paw, which are
cheap and environmentally safe, are promising for
protecting African yam bean seeds against major
seed-borne fungi. Many of the herbs properties
can be traced back to the flavonoids that plant
contains. Similarly, Olive leaves contain
oleuropein, eleonic Acid and other qualities that
can be of benefit to treat humans dermatophytes.
In this research, the percentage of inhibition (Table
4) was calculated after comparing with
Griseovolvin (100% inhibition). The maximum
activity was obtained from lemon grass which was
ranged from 75-95%, followed by lantana extract
which inhibited the fungal growth by 50-80% and
nerium and basil extract decreased growth by 30-
50%. The lowest activity was recorded for olive
leaves which inhibited the growth by 20-33.3 %.
The activity index was calculated. It was ranged
from 65-69% for both lemon grass and lantana,
38-39% for basil and nerium and 27% for olive
MICs of the six plant extracts were
calculated by using flurocin diacetate method
(Table 5). It was ranged from 1.0-1.5 µg/ml for
Griseofulvin. The MIC for the different plant
extracts were ranged from 25-75 for both lemon
grass, lantana and basil and from 100-175 µg/ml
for nerium and olive extract. The antifungal
activities of griseofulvin were determined by
Araújo et al. (2009) using broth microdilution
technique, against dermatophytes and the minimal
inhibitory concentrations (MICs) for Trichophyton
mentagrophytes, T. rubrum and Microsporum
canis were ranged from 0.03-1 µg/ml. It can be
concluded that, MICs calculated were greater than
that obtained for Griseofulvin. Further studies are
needed to determine the antifungal compound(s) in
such plant extract (isolation, separation and
identification) as well as its formulation to be
applicable as alternative methods to be used in
treatment of skin and skin structures diseases in
human and animal. Therefore, such results are of a
significant value that confirms the therapeutic
potency of some plants used in traditional
medicine. It should form a good basis for further
phytochemical and pharmacological investigation
(Prasad et al., 2009). Useful antimicrobial
phytochemicals are: phenolics and polyphenols
(such as simple phenols and phenolic acids,
quinones, flavones, flavonoids, and flavonols.
tannins, coumarins); terpenoids and essential oils;
alkaloids; lectins and polypeptides; plus other
compounds. The mechanisms thought to be
responsible for these phytochemicals against
microorganisims vary and depend on these
compounds (Aly and Bafiel, 2008). Their
mechanism of actions may include enzyme
inhibition by the oxidized compounds, and act as a
source of stable free radical and often leading to
inactivation of the protein and loss of function.
They have the ability to complex with extracellular
and soluble proteins and to complex with bacterial
cell walls and disrupt microbial membranes (Ali,
1999), some have ability to intercalate with DNA,
formation of ion channels in the microbial
membrane, competitive inhibition of adhesion of
microbial proteins to host polysaccharide receptors
The ultimate conclusion of this study
supports the traditional medicine use of different
plant extracts in treating different infections
caused by pathogenic fungi in Saudi Arabia either
by using a single or combined extracts. It also
suggests that a great attention should be paid to
medicinal plants which are found to have plenty of
pharmacological properties that could be
sufficiently better when considering a natural food
and feed additives to improve human and animal
Mycopath (2009) 7(1): 51-57
Antifungal activity of some medicinal plants 55
Table 1: Common and scientific names of some plants used to detect their antifungal activities in vitro.
Used part Family Scientific name Common name
Stalk and leaves Gramineae Oymbopogon citrates Lemon grass
Leaves and flower Verbenaceae Lantana camara Lantana
leaves Apocynaceae Nerium oleander Nerium
Stem and leaves Labiate
leaves Oleaceae Olea europaea Olive
Table 2: The % of fungal inhibition of aqueous and organic extract of different plants at concentration
100 µg/ml compared to Griseofulvin (100% inhibition) against Trichophyton rubrum.
Type of the extract chloroform n-butanol Ethyl
80 84 85 80 77 66 90 Lemon grass
60 80 85 80 32 44 88 Lantana
30 67 80 80 42 32 85 Nerium
80 57 73 44 30 24 73 Basil
77 55 75 67 30 26 75 Olive
The results were compared with that obtained for Griseofulvin which considered 100% inhibition.
Table 3: The antifungal activity of methanolic extract (diameter of the inhibition zone, mm) of different
plant extracts against different pathogenic fungi.
Diameter of the inhibition zone (mm) olive Basil Nerium Lantana Lemon
10 ±0.6 12 ±0.7 15 ±0.7 20 ±0.9 30 ±1.5 40 ± 2.5 Microsporum canis
10±0.5 15 ±0.6 14 ±0.8 20 ±0.7 22 ±0.5 40 ± 1.5 Microsporum gypseum,
10 ±0.4 16 ±0.5 16 ±0,6 20 ±0.6 38 ±1.4 40 ± 0.9 Trichophyton
8 ±0.5 20 ±0.3 20 ±0.9 20 ±1.5 30 ±1.6 40 ±0.9 Trichophyton verrucosum
10± 0.4 12 ±0.5 14 ±0.4 18 ±0.9 30 ±1.5 38 ±0.5 Epidermophyton floccosum
9.5 15 16 19.5 30 39.5 Activity Index*
GSF: Griseofulvin, *Activity index was calculated as the mean value of net zones of inhibition (mm)
against the five fungal test strains
Table 4: The % inhibition of the of different plant extracts compared to Griseofulvin (100% inhibition)
against different dermatophytes. Inhibition compared to Griseofulvin% olive Basil Nerium Lantana Lemon
25.0 30.0 37.5 50.0 75.0 100 Microsporum canis
25.0 37.5 35.0 80.0 55.0 100 Microsporum gypseum,
33.3 42.4 33.3 66.6 95.0 100 Trichophyton rubrum
20.0 50.0 50.0 75.0 75.0 100 T. verrucosum
26.3 31.5 36.0 78.9 79.0 100 Epidermophyton floccosum
27.0 39 38.0 69.0 65.0 100 Activity Index
*Activity index was calculated as the mean value of net zones of inhibition (mm) against the five fungal
Mycopath (2009) 7(1): 51-57
56 Fardos M. Bokhari
Table 5: Minimal inhibitory concentration (MIC) µg/ml of different plant extract using Fluorescein
diacetate method and compared with Griseofulvin.
Minimal inhibitory concentration (MIC)
grass lantana Nerium Basil Olive
Microsporum canis 1.5 ± 0.3 25 ± 3.6 50 ± 6.1 100 ±11.5 50 ±7.0 175 ± 3.0
Microsporum gypseum 1.0 ±0.1 25 ± 4.4 50 ± 4.1 100 ±14.0 50 ±6.0 175 ±7.8
Trichophyton rubrum 1.5 ± 0.5 25 ± 2.9 75 ± 7.0 100 ±8.0 125 ±11.0 150 ±12.0
T. mentagrophytes 1.5 ± 0.7 25 ± 5.2 50 ±5.0 125 ±6.3 50±3.0 100 ±5.9
T. verrucosum 1.0 ± 0.4 50 ± 3,9 50 ±4.4 125±11.1 75±4.2 125 ±13.0
GSF: Griseofulvin , All the values given, are the mean value of three reading.
The author appreciates the technical assistance of Dr. Magda Mohamed Aly, Biology Department, Faculty
of Science, King Abdel Aziz University, in this research
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