Sulaiman Ali Al Yousef
Sulaiman Ali Al Yousef
Sulaiman Ali Al Yousef
ABSTRACT
Recently, the use of essential oils against fungal infection has gained highly
importance because of acquired resistance against a large number of drugs. The
aim of study was testing the biological activity of essential oil vapors against some
Aspergillus spp., estimating chemical composition, active ingredient and
Keywords mechanism of action of tested oils. Lemongrass and peppermint essential oil
Lemongrass; volatiles were preliminary evaluated via vapor phase using volatilization method.
peppermint; Growth inhibition, minimum inhibitory concentration (MIC), and minimum lethal
antifungal concentration (MLC) of these pathogenic fungi were used to detect their
activity; susceptibility to each of the essential oils. Morphological changes of treated fungi,
Aspergillus as well as its spores germination was observed by light and electron microscopy.
Also, chemical analysis of essential oils were achieved by GC, GC-MS.
spp.;
Lemongrass oil was found to be highly fungicidal, as it showed the lowest MIC and
essential oils. MLC values and the highest growth inhibition; in a range of concentrations (15 to
20 l/0.4l air space) it was effective in inhibiting fungal viability and spore
germination. The main morphological changes caused by Lemongrass oil in A.
niger were observed under both the light and electron microscope; these included a
reduction in conidition, loss of pigmentation and disrupted conidispore structure.
The results show that Lemongrass oil produces a fungitoxic effect, which supports
its possible use in medicine to cure mycotic infections.
Introduction
Many of the plant species found that it can 2005) activity. Lemongrass (Cymbopogon
be used medically (Ali-Shtayeh and Abu, citratus) is widely used in traditional
1999). Essential oils are aromatic medicine in many countries around the
substances which are obtained from world. Among its attributable properties
various plant parts by steam distillation; are those related to antibacterial and
many of which exhibit antibacterial antifungal activities (Inouye et al., 2001,
(Ozcan et al., 2006), antifungal (Chee and Inouye et al., 2006; Bansod and Rai, 2008;
Lee, 2007) and antiviral (Khan et al., Revathi et al., 2012), as well as analgesic
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Doses of the essential oils were applied by Assessment of the effect of oil volatiles
gaseous contact as described above in on fungal morphology
order to determinate their potential to
inhibit fungal spore germination A spore Morphology alterations caused by sub-
suspension (300 l harvested from a 7 day lethal dose of lemongrass oil against A.
old culture (106 spores/ml) was spread on niger were observed in duplicate by light
glass slides which were incubated in an and electron microscope. According to
atmosphere of the essential oils at 30C for The following the slide culture technique
24 hours. At the end of incubation period, was performed (Pereira, et al, (2011). A
each slide was fixed with lacto-phenol- small block of Czapek-Dox agar was
cotton blue stain and observed under the transferred to a glass slides in a Petri dish
light microscope for spore germination; and then inoculated with the test fungus.
approximately 150 spores were counted. The required dose was pipetted onto a
filter paper disc as described above and the
Assessment if minimum inhibitory plate was incubated at 30C for 5 days.
concentration (MIC) and minimum Control assays lacking the essential oil
lethal concentration (MLC) of oil under test were included. After incubation,
volatiles the slides were observed under a
stereomicroscope at 200X. In E/M studies,
In order to demonstrate the MIC and MLC
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Table.1 the chemical composition of the essential oil from Lemongrass and
Peppermint analyzed by GC-MS
Lemongrass Peppermint
Component % Conc. Component % Conc.
pinene 0.06 thujene 0.04
cis-sabinene hydrate 0.11 Linalool 0.72
1-8 cineole 0.12 1,8-Cineole 8.15
Geranyl acetate 0.23 Menthyl acetate 5.28
Geraniol 0.72 Isomenthone 9.47
Terpinolene 1.52 Myrcene 0.45
-caryophyllene 2.05 Limonene 1.8
Linalool 3.82 trans-sapinin hydrate 1.42
Limonene 4.12 Sabinene 0.91
3-myrcene 18.5 -pinene 1.05
Neral* 23.92 Menthone 28.46
Geranial* 46.25 Menthol 24.5
*a mixture of Neral and geranial is called Citral
Table.2 Antifungal activity of lemongrass and peppermint essential oil against
Aspergillus spp. growth in vitro.
Means in each column having the same superscript letters are not significantly different a
p 0.05*Results expressed in percent of spore germination inhibition in comparison
with the control assay
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Table.4 MIC and MLC of response to volatiles of oil of Lemongrass and Peppermint
against Aspergillus spp.
Means in each column having the same superscript letters are not significantly
different a p 0.05
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and Kunicka, (2003) and Hamza et al., chrysogenum, Mucor fragilis, and
(2009), have also demonstrated synergistic Rhizopus stolonifer (Duarte et al., 2005;
effects of the different compounds which Inouye et al,, 2007; Agarwal et al., 2008;
are present in natural essential oils; our van Vuuren et al., 2009; Yigit et al.,
results are therefore in agreement with 2009). Then, antimicrobial activity of the
previous findings. peppermint oil samples decreased in the
order of total content of 1,8-cineole,
Previous investigations on M. piperita oil menthol and -caryophyllene decreasing
composition are consistent with our results (Gochev et al., 2008).
in which menthol and menthone were
found to be the major fungus-inhibitory Microscope studies on the susceptibility of
compounds (Rohloff, 1999; Maffei, 1999; A.niger, to Lemongrass show that the
Gerherman et al., 2000; Aflatuni et al., fungus underwent major morphological
2000). Based on the findings from a changes induced by lemongrass stress
variety of laboratory studies, Peppermint (Figure 2) which were generally consistent
oil and menthol have been shown to with the results of earlier publications and
possess action against a variety of fungi, confirm the fungicidal effect of
including Aspergillus niger, A. flavus, A. Lemongrass. Earlier studies have also
fumigatus, A. sulphureus, A. austriaca, demonstrated the inhibitory effect of
Candida spp., Fusarium spp, lemongrass on hyphal growth and spore
Trichophyton spp., Penicillium formation in A. niger, as well as plasma
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