ArticlePDF Available

Chemical composition, antiviral against avian influenza (H5N1) virus and antimicrobial activities of the essential oils of the leaves and fruits of Fortunella margarita, lour. swingle, growing in Egypt

Authors:

Abstract and Figures

Essential oils of the fresh leaves and fruits of Fortunella margarita Lour. Swingle (Family: Rutaceae) were prepared by hydrodistillation method, which resulted with 0.27 and 0.30% respectively. The resulted oils of both organs were analyzed by GC/MS which revealed the presence of twenty compounds in the leaves oil representing 86.96% of the oil, from which gurjunene, eudesmol and muurolene were identified as major compounds. The fruit's oil was found to contain fourteen compounds representing 77.77% of the oil, of which terpineol, t-carveol, limonene, muurolene and cadinene represented the major compounds. The antiviral activity of the essential oils of both leaves and fruits was tested against avian influenza-A virus (H5N1), and the results revealed higher potency of fruits oil. Moreover, the essential oils of the leaves and fruits were investigated for their antimicrobial and antifungal activities. The oil of the leaves showed antimicrobial activity higher than that of the fruits at dilution (1:50 v/v) against Bacillus subtilis, Staphylococcus aureus, Sarcina luta and Streptococcus faecalis, also it has a moderate activity against Escherichia coli, Klebsilla pneumonia and Pseudomonas aeroginosa. On the other hand, the antifungal activity of the leaves and fruits revealed that the fruits exhibited higher activity than that of the leaves against Aspergillus niger and Candida albicans.
Content may be subject to copyright.
© 2015 Nabaweya A. Ibrahim et al. This is an open access article distributed under the terms of the Creative Commons Attribution License -NonCommercial-
ShareAlikeUnported License (http://creativecommons.org/licenses/by-nc-sa/3.0/).
Journal of Applied Pharmaceutical Science Vol. 5 (01), pp. 006-012, January, 2015
Available online at http://www.japsonline.com
DOI: 10.7324/JAPS.2015.50102
ISSN 2231-3354
Chemical Composition, Antiviral against avian Influenza (H5N1)
Virus and Antimicrobial activities of the Essential Oils of the Leaves
and Fruits of Fortunella margarita, Lour. Swingle, Growing in Egypt
Nabaweya A. Ibrahim1, Seham S. El-Hawary2, Magdy M. D. Mohammed1,*, Mohamed A. Farid3, Nayera A. M. Abdel-
Wahed3, Mohamed A. Ali4, Eman A. W. El-Abd1
1 Pharmacognosy Department, Pharmaceutical and Drug Industries Research Division, National Research Center, Dokki-12311, Cairo, Egypt.
2 Pharmacognosy Department, Faculty of Pharmacy, Cairo University, Giza, Egypt. 3Chemistry of Natural and Microbial Products Department,
Pharmaceutical and Drug Industries Research Division, National Research Center, Dokki-12311, Cairo, Egypt. 4Center of Scientific Excellence for
Influenza Viruses, National Research Center, Dokki-12311, Cairo, Egypt.
ARTICLE INFO
ABSTRACT
Article history:
Received on: 28/10/2014
Revised on: 10/11/2014
Accepted on: 05/12/2014
Available online: 30/01/2015
Essential oils of the fresh leaves and fruits of Fortunella margarita
Lour. Swingle (Family: Rutaceae) were
prepared by hydrodistillation method, which resulted with 0.27 and 0.30% respectively. The resulted oils of both
organs were analyzed by GC/MS whi
ch revealed the presence of twenty compounds in the leaves oil representing
86.96% of the oil, from which gurjunene, eudesmol and muurolene were identified as major compounds. The
fruit’s oil was found to contain fourteen compounds representing 77.77% of the oil, of which terpineol, t-
carveol,
limonene, muurolene and cadinene represented the major compounds. The antiviral activity of the essential oils
of both leaves and fruits was tested against avian influenza-A virus (H5N1), and the results revealed high
potency of fruits oil. Moreover, the essential oils of the leaves and fruits were investigated for their antimicrobial
and antifungal activities. The oil of the leaves showed antimicrobial activity higher than that of the fruits at
dilution (1:50 v/v) against Bacillus subtilis, Staphylococcus aureus, Sarcina luta and Streptococcus faecalis
, also
it has a moderate activity against Escherichia coli, Klebsilla pneumonia and Pseudomonas aeroginosa. On the
other hand, the antifungal activity of the leaves and
fruits revealed that the fruits exhibited higher activity than
that of the leaves against Aspergillus niger and Candida albicans.
Key words:
Fortunella margarita Lour.
Swingle; Rutaceae; essential
oil; antiviral; antimicrobial.
INTRODUCTION
Kumquats (Fortunella spp.) belong to the Citrus genus;
their fruits are usually eaten raw as a whole fruit together with
the peel, excluding the seeds. The peel is sweet and edible with a
typical aroma due to the presence of flavonoids and terpenoids
(Koyasako and Bernhard, 1983). Fortunella margarita Lour.
Swingle (Rutaceae) is an evergreen tree native to Southeastern
Asia and more precisely to China. It is also known as the oval or
nagami kumquat. The genus Fortunella has been used in folk
medicine to treat fevers, gallstones, indigestion, hernial pain,
.
* Corresponding Author
Dr. Magdy Mostafa Desoky Mohammed, Ph.D.
Department of Pharmacognosy, National Research Center, Dokki
-
12311, Cairo, Egypt. Tel.: +202-33371718; Fax: +202-33370931
E
-mail: melhenawy111@gmail.com (Magdy M. D. Mohammed).
stomachache, hepatitis, high blood pressure, prolapse of the uterus
and anus, asthma, catarrhal cough, pneumonia, respiratory
congestion and whooping cough (Khaleel et al., 2001). Certain
Fortunella species were reported to be used as haemostatic,
antiasthmatic and in treatment of diarrhea, Fortunella margarita
Lour. Swingle is cultivated in Egypt for ornamental purposes
(Khaleel et al., 2001). Kumquats are also an excellent source of
nutrients and phytochemicals, including ascorbic acid, carotenoids,
flavonoids and essential oils (Wang et al., 2012) i.e., 3´,5´-di-C-β-
glucopyranosyl phloretin, which is a characteristic flavonoid in F.
margarita and all other Fortunella species (Ogawa et al., 2001).
Some Fortunella species i.e., F. japonica (round), F. margarita
(oval or nagami) and F. crassifolia (jingdan or meiwa) are
commonly cultivated in the Southern region of China. Most of
Fortunella species can be used for the preparation of marmalade,
fruit salad and as food preservative.
Ibrahim et al. / Journal of Applied Pharmaceutical Science 5 (01); 2015: 006-012 07
As a result of inadequate use of antiviral drugs, influenza
viruses are mutating and mutant variants are evolving. To control
the spread of these viruses as well as the expected and unexpected
mutant variants, we have a great challenge to find out more potent
antiviral agents. Plant and marine extracts, synthetic compound
and target directed compounds are the main sources of these
antiviral agents.
Antiviral and antimicrobial drugs are subject to microbial
resistance, and this has become a growing public problem all over
the world. Therefore, ample research to discover potent new
antibiotics is compulsory. Since many essential oils have been
reported to possess strong antimicrobial effects (Nakatsu et al.,
2000, Rios and Recio, 2005, Koch et al., 2008), the aims of the
present study were conducted to investigate the chemical
composition of the essential oils of both leaves and fruits of
Fortunella margarita, then the antiviral activity against pathogenic
avian influenza virus (H5N1), and also the antimicrobial, and
antifungal activities were performed.
MATERIALS AND METHODS
Plant materials
Fortunella margarita fruits and leaves were collected
from Egypt green farm at Cairo-Alexandria agriculture road.
Leaves were collected at the flowering stage while fruits were
collected in February 2007. The plant was kindly identified by
Mrs. Theresa Labib, consultant of taxonomy at the ministry of
agriculture and the former director of El-Orman botanical garden.
Voucher specimen of the whole plant (000120FC 04-09-06-25)
was kept at the garden.
Essential oil isolation
The essential oils were extracted from fresh fruits and
leaves of F. margarita by hydrodistillation method (ShunZhen et
al., 2012). The oil content of each sample was determined as mean
of triplicate. The collected oils were subjected to GC/MS analysis.
Qualitative and quantitative identification of the oil constituents
were carried out by comparing the retention times and mass
fragmentation pattern with the previously published data (Adams,
1989, Walter and Takayuki, 1980).
Analysis of the essential oil
GC/MS conditions; the compounds were separated on an
HP-5-MS fused silica capillary column (30 m x 0.25 mm ID x 0.25
μm (film thickness), Agilent, Palo Alto, CA, USA).
Helium 5.0 was used as a carrier gas at a constant flow rate of 1.5
mL/min. The GC was operated in split less injection mode and
the PTV injector was programmed from 60 to 285 ºC
(1.1 min) at 14.5 ºC/s at an injection volume of 1 micro
Litter (Ligon et al., 2008). The Trace MS Plus detector was
operated in selected ion monitoring (SIM) mode at the
ionization energy of 70 eV. The transfer line between GC
and MS was kept at 250 ºC and the ion-source temperature
was kept at 200 ºC. The MS calibration was done by auto-tuning.
Antiviral Activity
Virus and cells
Reasserted avian influenza A virus (H5N1) previously
isolated from Egypt in 2006 (rgA/chicken/Egypt/1/2006), was
used in this study to evaluate antiviral activity of the studied
extracts. Madin-Darby canine kidney (MDCK) cells used for virus
propagation were friendly obtained from St. Jude Children’s
Research Hospital. The MDCK cells were routinely passaged in
Dulbecco’s modified Eagle medium (DMEM) containing 10%
fetal bovine serum and 1% antibiotic-antimycotic mixture
(penicillin- streptomycin-amphotericin B).
MTT assay (Cytotoxicity assay)
The stock samples were diluted with Dulbecco's
Modified Eagle's Medium (DMEM) to desired concentrations.
Stock solutions of the test compounds were prepared in DMSO at
a concentration of 10% in dH2O (distilled H2O). The cytotoxic
activity of the extracts were tested in MDCK cell line by using the
3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide
(MTT) method (Hayden et al., 1980, Mossman, 1983) with minor
modification. Briefly, the cells were seeded in 96-well plates (100
μL/well at a density of 3×105 cells/mL) and treated with various
concentrations of the sample solutions. After 24 h, cells were
washed with sterile phosphate buffer (PBS) 3 times and the
supernatant was discarded. MTT solution (20 μL of 5 mg/mL) was
added to each well and incubated at 37 °C for 4 h.
% Cytotoxicity =
(Absorbance of cell without treatment – Absorbance of cell with treatment) ×100
Absorbance of cell without treatment
Then the medium was aspirated. In each well, the formed
formazan crystals were dissolved with 200 μL of acidified
isopropanol (0.04 M HCl in absolute isopropanol). An absorbance
of formazan was detected by a dual wavelength UV spectrometer
at 540 nm with 620 nm reference wavelength. The percentage of
cytotoxicity compared to the untreated cells was determined with
the equation given above. The plot of % cytotoxicity versus
sample concentration was used to calculate the concentration
which exhibited 50% cytotoxicity (TC50).
Microorganisms
Bacterial strains
Five Gram-positive bacteria strains viz; (Bacillus subtilis
NRRL 543, Staphylococcus aureus NRRL B-313, Sarcina luta
NRRL B-1018, Streptococcus faecalis NRRL 537, Arthobacter
citreus NRRL B-1258) and three Gram-negative bacteria strains
viz; (Escherichia coli NRRL B-210, Klebsilla pneumonia NRRL
B-117 and Pseudomonas aeroginosa NRRL B-23) were obtained
from both of the Northern Utilization Research and Development
Division, United State Department of Agriculture, Peoria, Illinois,
USA and the Department of Microbiology, Faculty of pharmacy
Cairo university, Egypt. The bacterial strains were revived for
bioassay by sup-culturing in fresh nutrient broth medium for 24
hours before test.
08
Ibrahim
et al. /
Journal of Applied Pharmaceutical Science 5 (01); 2015: 006-012
Fungal strains
The tested fungi including Aspergillus niger NRRL-599
and Candida albicans NRRL Y-477 were cultured on potato
dextrose agar (PDA) (2.5% agar) for 7 days at 28 °C before the
experiment was carried out.
Preparation of plates
The antibacterial and antifungal tests of the essential oils
were tested using agar diffusion method (Linday, 1962), the ready-
made nutrient agar medium (for bacterial strains) and PDA (for
fungal strains) were suspended in distilled water and autoclaved at
pressure of 1.5 atm. For bacterial strains a suspension of tested
microorganisms (0.1 mL of about 106 cells/mL) was seeded on
solid media plates (9 cm diameter) and uniformly spread with a
sterile spreader. Six to eight wells (8 mm) were made on the solid
medium with a sterile cork borer. To each well, fixed volume (0.1
mL) of diluted essential oil and carefully was placed in each well.
For fungal strains seven days old cultures of test organisms
(0.1 mL) were used. Controls were maintained with paraffin oil
only.
Antimicrobial activity of the oils
The antibacterial and antifungal tests of the essential
oils were tested using the agar diffusion method (Linday, 1962).
The treated and the controls were kept in an incubator at
37 °C for bacterial strains and at 28 °C for 24 to 72 hours. The
zones of inhibition for each well were measured.
Nystatin, erythromycin, oxacillin, methcilin and bacitracin were
included in the test as references. At the end of incubation
period, the diameter (mm) of the inhibition zones was
measured. Plates were done in triplicate and an average + SD was
recorded.
The minimum inhibitory concentration (MIC)
The stock solutions of the oils were diluted and
transferred into the first tube, and serial dilutions were performed
so that concentrations in the range of 0.001-0.02 µL/mL were
obtained. A 10 µL spore suspension of each test strain was
inoculated in the test tubes in nutrient medium and incubated for
24-72 hours at 37 °C. The control tubes containing the same
medium were inoculated only with bacterial strains suspension.
The minimal concentrations at which no visible growth was
observed were defined as the MICs which were expressed in (v/v
%).
Spore germination assay
Spore germination assay was carried out according to
(Rana et al., 1997). Three concentrations (4, 2, and 1% v/v) of
each oil sample, together with two controls (one sterile distilled
water and other of 0.1 % (v/v) methanol in sterile distilled water)
were tested for spore germination of Aspergillus niger and
Fusarium oxysporum.
Aliquots of 0.1 mL from each sample were mixed with
fungal spores obtained from 10 days old cultures of the tested
fungi and placed on separate glass slides in triplicate. Slides
containing the spores were incubated in a moist chamber at 28 °C
for 24h. Each slide was then fixed in lactophenol-cotton blue and
observed under the microscope for spore germination.
RESULTS AND DISCUSSION
Essential oils of the fresh leaves and fruits of Fortunella
margarita Lour. Swingle (Family: Rutaceae) were prepared by
hydrodistillation method, and resulted with a mean percentage of
three replicates of 0.27 and 0.30 % respectively (Table 1). The oils
of both organs were analyzed by GC/MS using the previously
mentioned conditions, which revealed the presence of twenty
compounds in the leaves representing 86.96% of the total leaves
oil, from which the major compounds were eudesmol (36.66%),
muurolene (10.26%), and gurjunene (9.98%).
The oil of the fruits were found to contain fourteen
compounds representing 77.77% of the total fruits oil, terpineol
(55.47%), t-carveol (5.51%), limonene (1.67%), muurolene
(5.51%) and cadinene (2%) represented the major compounds.
Terpineol is a naturally occurring monoterpene alcohol that has
been isolated from a variety of sources such as cajuput oil, pine
oil, and petitgrain oil (Merck Index). There are three isomers,
alpha-, beta-, and gamma-terpineol, the last two differing only by
the location of the double bond. Terpineol is usually a mixture of
these isomers with alpha-terpineol as the major constituent. It has
a pleasant odor similar to lilac and is a common ingredient in
perfumes, cosmetics, and flavors. α-Terpineol is one of the two
most abundant aroma constituents of lapsang souchong tea; the α-
terpineol originates in the pine smoke used to dry the tea (Yao et
al., 2005).
Previous reports by ShunZhen et al., (2012) of the
essential oil composition isolated from the leaves and fruit peels of
F. margarita revealed the presence of 27 compounds in the leaves
with 91.37%, while 34 compounds were identified from fruit peels
sample with 96.23%. The main components of both organs were
different, except only 13 components i.e., α-pinene, linalool, β-
caryophyllene etc., were the same but with different content.
On the other hand the volatile compositions of three
varieties of kumquats; F. crassifolia, F. margarita and F. japonica
were determined by using headspace solid phase micro-extraction
(HS-SPME) coupled with GC-MS. Twenty-eight components in
the three tested kumquat varieties were identified, with D-
limonene is the major component with 50.40%, 55.43% and
51.47%, respectively.
Furthermore, α-pinene, D-limonene, cis-α-cadinene, δ-
cadinene, iso-caryophyllene, β-elemene and acetic acid esters were
common in all samples. In addition, each kumquat variety contains
its peculiar components, i.e., F. crassifolia Swingle contains 4-
carene, F. margarita Swingle contains D-germacrene, cedrene and
linalool and F. japonica (Thunb.) Swingle contains limonene,
ocimene, 2,6,10,14-tetramethyl-heptadecanoic, cuba ene, 2,6-di-
tert-butyl-4-butyl phenol and isopropyl palmitate (Zhonghai et al.,
2009).
Ibrahim et al. / Journal of Applied Pharmaceutical Science 5 (01); 2015: 006-012 09
Fig.1 Gas ion chromatogram of the essential oil of Fortunella margarita fruits.
Fig. 2 Gas ion chromatogram of the essential oil of Fortunella margarita leaves
010
Ibrahim
et al. /
Journal of Applied Pharmaceutical Science 5 (01); 2015: 006-012
Table 1: GC/MS of the essential oil of F. margarita fruits and leaves
Peak no. Rt Components RRt (min.) of
fruits Conc.% of
fruits RRt (min.) of
leaves Conc.% of leaves
Monoterpenes
1 5.11 Linalool 0.98 0.84 - -
2 5.17 Unknown 0.99 5.51 - -
3 5.22 α-Terpineol 1 55.47 - -
4 5.38 Carveol 1.03 5.51 - -
5 5.55 Carvone 1.06 5.68 - -
6 5.78 Limonene 1.1 1.67 - -
7 6.21 Dihydrocarveol - - 0.69 2.4
8 6.41 Citronellal 1.23 5.01 - -
Sesquiterpenes and others
9 6.5 α-Cubebene - - 0.72 0.07
10 6.57 δ-Cadinene 1.25 0.67 - -
11 6.6 β-Bourbonene - - o.73 2.89
12 6.66 Bisabolene trans-gamma 1.28 0.67 - -
13 6.85 δ-Elemene - - 0.76 5.32
14 6.91 β-Cubebene - - 0.77 0.6
15 6.95 α-Gurjunene - - 0.77 0.37
16 7.1 Bisabolene - - 0.79 0.89
17 7.26 γ-Muurolene 1.39 5.51 - -
18 7.33 γ-Muurolene - - 0.82 6.53
19 7.33 Germacren B 1.4 1.75 - -
20 7.38 α-Guaiene - - 0.82 1.26
21 7.41 α-Humulene - - 0.83 2.41
22 7.51 α-Cadinene 1.44 1.34 - -
23 7.54 γ-Cadinene - - 0.84 1.55
24 7.67 γ-Eudesmol - - 0.85 0.6
25 7.88 Unknown - - 0.88 7.71
26 7.96 Unknown - - 0.89 1.88
27 8.05 Diethylphthalate 1.54 2.26 - -
28 8.14 δ-Cadinene - - 0.9 1.29
29 8.24 Unknown - - 0.91 3.01
30 8.3 Cadinol - - 0.92 2.04
31 8.34 Unknown 1.6 2.34 - -
32 8.38 β-Cedrene - - 0.94 1.89
33 8.64 Unknown 1.66 2.51 - -
34 8.7 β-Gurjunene - - 0.97 9.98
35 8.72 α-Muurolene - - 0.97 10.26
36 8.82 Cedrol 1.69 1.5 - -
37 8.89 β-Eudesmol - - 1 28.25
38 9.02 γ-Eudesmol - - 1.004 8.41
39 9.11 Germacren D-4-ol - - 1.02 0.42
40 10.72 Unknown 2.05 0.75 - -
41 11.7 Galaxolide 1.24 1 - -
RRt: Relative Retention Time to α-Terpineol in Fruits and to β-Eudesmol in Leaves
Table 2: Antiviral activity of F. margarita fruits and leaves.
Oils TC50 (µg/mL) H5N1
Initial Viral Count PFU/ml Conc. Of Sample(µg) % of reduction IC
50
(µg/mL)
F. margarita fruits 239.54
8.2 X 105
5 60.97 6.77 10 70.73
20 78.04
F. margarita leaves 185.47 5 39.02 38.89 10 41.46
20 43.9
TC50: It is the Half Maximal (50%) Toxic Concentration of the Sample on Cell Line under Examination.
PFU: Plaque Forming unit: Refers to One Infectious Virion that Is Capable of Initiation of Infection in One Cell Rises into a Plaque after Incubation.
IC50: It Is the Half Maximal (50%) Inhibitory Concentration (IC) of the Sample Reducing
Table 3: MICs of the essential oils from F. margarita fruits and leaves.
Test organism MIC (% v/v) Oil of fruits MIC (%v/v) Oil of leaves
1- Bacillus subtilis
2- Escherichia coli
3- Aspergillus niger
4- Candida albicans
2
1
0.01
0.01
0.01
ND
0.01
0.01
ND: Not Determined
Ibrahim et al. / Journal of Applied Pharmaceutical Science 5 (01); 2015: 006-012 011
Antiviral activity
Avian influenza (AI) is a highly contagious disease of
poultry caused by influenza viruses type-A of the family
Orthomyxoviridae. The rapid rate of spread and the high potential
for genetic alterations of the virus has raised the spectra of wide
spread human infection and the possibility of a pandemic. With
emergence and multi-regional spread of AI especially highly
pathogenic avian influenza (HPAI) of H5 and H7 subtypes in
poultry, the epizootic has altered the world to the prospects of a
potentially devastating human health challenge. The HPAI (H5N1)
subtype virus has caused disease outbreaks in poultry in several
countries in Asia including India. The role of antivirals is
considered critical in preparedness for avian flu originated
pandemic. Several at-risk nations and WHO have stored strategic
stockpiles of antivirals especially oseltamivir to be used at the face
of influenza pandemic. However, resistance to oseltamivir in the
H5N1 subtype in Vietnam and other human influenza A viruses
has become a cause for worry as far as pandemic preparedness is
concerned. Therefore, searching for alternatives for antivirals that
can effectively inhibit H5N1 or other influenza A viruses, and/or
act in synergy with available antivirals, is an urgent need of the
hour. Several novel antiviral agents that may be effective against
influenza virus especially H5N1 avian flu virus, are currently
under development. Among these, are plant-derived extracts,
which have become the focus of many studies due to their proven
beneficial health effects in several disease problems (Sood et al.,
2012). In the present study the essential oils of both organs
(leaves and fruits) of F. margarita were tested for the first time for
their antiviral activity against avian influenza (H5N1) virus, the
obtained results (Table 2) revealed that the fruits essential oil was
more effective (80% virus inhibition) that of the leaves, this was
attributed to the presence of α-terpineol as a major component in
fruits oil. Our obtained results comes in accordance with the
previous study by (YaDong et al., 2009) which investigated the
antiviral effect of Curcuma zedoaria volatile oil and Hypericin
perforatum liquid extract on H5N1 avian influenza virus (AIV) in
MDCK cell line and non-AIV-immunized chickens, and attributed
the virucidal activity to the presence of curcumenol and hypericin.
Antimicrobial and Antifungal activity
The essential oils of both organs (fruits and leaves)
showed antimicrobial properties (Table 3) with regard to their
inhibition zone (IZ) ranging from 16 to 31 mm. Fruits oil was
active against Gram +ve, Gram -ve, yeast and fungi. The diameter
of inhibition zone was ≥25mm at 100 µL oil concentration against
Streptococcus faecalis, Klebsilla pneumonia, Pseudomonans
aeroginosa, Asperigillus niger, and Candida albicans.
Asperigillus niger, and Candida albicans were the most
susceptible microorganisms inhibited by fruits oil, followed by the
Gram +ve bacteria Streptococcus faecalis, Gram -ve bacteria
Klebsilla pneumonia, Pseudomonans aeroginosa. However, the
fruits oil being less active (IZ < 25) against Bacillus Subtilis,
Staphylococcus aureus, Sarcina luta, Anthrobacter and
Escherishia coli. Moreover, the leaves oil showed significant
activity against all tested microorganisms except Streptococcus
faecalis and Klebsilla pneumonia, with inhibition zone 25
observed for Bacillus subtilis, Sarcina luta, Staphylococcus
aureus. This can be attributed to the presence of eudesmol as a
major component of leaves oil which was reported to have a strong
Table 4: Antimicrobial activities of the essential oils from F. margarita fruits and leaves.
Inhibition zone
diameter (mm)
Sample
Fruits oil(a) Leaves oil(a) Erythromycin 150
µg/mL Methicillin 50
µg/mL Oxacillin 10
µg/mL Bacitracin
100 µg/mL Nystatin 100
µg/mL
Gram +ve bacteria
Bacillus subtilis 21 31 26 14 -ve 18 11
Staphylococcus aureus 22 28 16 -ve -ve 12 12
Sarcina luta 20 32 25 24 24 26 -ve
Streptococcus faecalis 25 -ve 20 -ve -ve 19 -ve
Anthrobacter 18 19 28 19 24 12 12
Gram -ve bacteria
Escherichia coli 16 16 16 11 12 12 -ve
Klebsilla pneumonia 25 -ve -ve 12 14 -ve -ve
Pseudomonas aeroginosa 25 19 35 17 15 12 -ve
Yeast and Fungi
Aspergillus niger 29 25 -ve -ve -ve -ve 15
Candida albican 29 17 -ve -ve -ve -ve 15
a: 0.1 mL of Diluted Essential Oil (1:50 v/v)
Table 5: Effect of essential oils from F. margarita fruits and leaves on spore germination of A. niger and F. oxysporum.
Essential oil Conc. of oil (%v/v) Aspergillus niger Fusarium oxysporum
Oil of fruits 4
2
1
-ve
-ve
-ve
-ve
-ve
+ve
Oil of leaves 4
2
1
+ve
+ve
+ve
+ve
+ve
+ve
+ve Effect: Refers to Germination of the Fungal Strain after 24 and 48 Hours
012
Ibrahim
et al. /
Journal of Applied Pharmaceutical Science 5 (01); 2015: 006-012
antimicrobial and free radical scavenging activities (Amezouar et
al., 2012). To the best of our knowledge the IZ for both oils were
higher than that of the reference drug.
The minimum inhibitory concentrations of both oils of
Fortunella margarita obtained by the agar diffusion method are
shown in (Table 4). Both oils inhibited Asperigillus niger and
Candida albicans at 0.01% v/v. Fruits oil inhibited Bacillus
Subtilis and Escherishia coli at 2 and 1% respectively, while
leaves oil inhibited Gram +ve bacteria Bacillus Subtilis at 0.01
%v/v. The results in (Table 5) revealed that the fruits oil
completely inhibited the germination of Aspergillus niger and
Fusarium oxysporum spores at all concentrations tested except for
Fusarium oxysorum at 1% (v/v). On the other hand, leaves oil has
no inhibitory effect on seed germination of the tested fungal
strains. A separate two control run simultaneously in the present
study showed that it did not inhibit spore germination.
The mode of action of antimicrobial action of essential
oil may be due to inhibition of respiration and disrupting the
permeability barriers of the cell membrane structures (Cox et al.,
2000).
CONCLUSION
The biological activity of these essential oils is due to the
presence of synergistic effect of mixture of volatile compounds or
their major constituents. The strong antiviral activity against
pathogenic avian influenza (H5N1), and the broad spectrum of
antimicrobial and antifungal activities of the essential oils against
variety of bacterial and fungal strains so we can recommend that
the essential oil of F. margarita can be incorporated in different
pharmaceutical preparations for the first time.
REFERENCES
Adams R.P. (1989). Identification of the Essential Oils by Trap
Mass Spectroscopy. Academic press, INC, London.
Amezouar F., Badri W., Hsaine M., Bourhim N., Fougrach H.
Chemical composition, antioxidant and antibacterial activities of leaves
essential oil and ethanolic extract of Moroccan Warionia saharae Benth.
& Coss. J. Appl. Pharm. Sci. 2012; 02(05), 212-217.
Cox S.D., Mann C.M., Markham J.L. The mode of
antimicrobial action of the essential oil of Melaleuca alternifolia (tea tree
oil). J. Appl. Microbiol. 2000; 88, 170-175.
Hayden F.G., Cote K.M., Douglas R.G. Plaque inhibition assay
for drug susceptibility testing of influenza viruses. Antimicrobial Agents
Chemother.1980; 17, 865–870.
Khaleel A.E., El-Gayed S.H., Nolte J., Fobbe R. Comparitive
study of the essential oils of the leaves and fruits of Fortunella margarita
(Lour.) Swingle grown in Egypt. Al-Azhar J. Pharm. Sci. 2001; 28, 258-
266. Koch C., Reichling J., Scheele J., Schnitzler P. Inhibitory effect
of essential oils against Herpes simplex virus type 2. Phytomedicine,
2008; 15, 71-78.
Koyasako A., Bernhard R.A. Volatile constituents of essential
oils of kumquat. J. Food Sci. 2002; 48, 1807–1810.
Ligon A.P., Zuehlke S., Spiteller M. GC-MS analysis of organic
compounds in waste water and sewage sludge. J. Sep. Sci. 2008; 31, 143
– 150. Linday E.M. (1962). Practical Introduction to
Microbiology. (E and FN spon Ltd), UK. pp77. Merck Index, 11th Edition,
9103. Mossman T. Rapid colorimetric assay for cellular growth and
survival: application to proliferation and cytotoxicity assays. J. Immunol.
Methods, 1983; 65, 55-63.
Nakatsu T., Lupo A.T. Jr, Chinn J.W. Jr, Kang R.K.L. (2000).
Biological activity of essential oils and their constituents. In, Studies in
Natural Products Chemistry; Bioactive Natural Products (Part B). Edited
by Atta-ur-Rahman. Vol. (21), 571-631.
Ogawa K., Kawasaki A., Omura M., Yoshida T., Ikoma Y.,
Yano M. 3´,5´-Di-C-β-gluco- pyranosyl phloretin, a flavonoid
characteristic of the genus Fortunella. Phytochemistry, 2001; 57, 737–742.
Rana B.K., Singh U.P., Taneja V. Antifungal activity and
kinetics of inhibition by essential oil isolated from Aegle marmelos. J.
Ethnopharmacol. 1997; 57, 29-34.
Rios J.L., Recio M.C. Medicinal plants and antimicrobial
activity. J. Ethnopharmacol. 2005; 100, 80-84.
ShunZhen L., HongXing L., LiXia Z., YanMin H. Comparative
analysis on chemical components in essential oils from the
leaves and fruit peels of Fortunella margarita. Medicinal Plant. 2012; 3,
43-46. Sood R., Swarup D., Bhatia S., Kulkarni D.D., Day S., Saini M.,
Dubey S.C. Antiviral activity of crude extracts of Eugenia jambolana
Lam. against highly pathogenic avian influenza ( H5N1) virus. Indian J.
Exp. Biol. 2012; 50, 179-186.
Walter J., Takayuki S. (1980). Quantitative analysis of flavor
and fragrance volatile by Glass Capillary Gas Chromatographay.
Academic Press, INC, New York. Wang Y.-W., Zeng W.-C., Xu P.-Y.,
Lan Y.-J., Zhu R.-X., Zhong K., Huang Y.-N., Gao
H. Chemical composition and antimicrobial activity of the
essential oil of Kumquat (Fortunella crassifolia Swingle) Peel. Int. J. Mol.
Sci. 2012; 13, 3382 3393.
YaDong H., YanMei L., Qi X., ZhiJian S., LinChuan W.,
XiaoKun L., ZhiFeng H. Antiviral effect on H5N1 avian influenza virus
of compound of Curcuma zedoaria volatile oil and
Hypericin perforatum extract liquid. J. China Pharma Univ. 2009; 40 (2),
166-172. Yao S.-S., Guo W.-F., Lu Y., Jiang Y.-X. Flavor characteristics
of lapsang souchong and smoked lapsang souchong, a special Chinese
black tea with pine smoked process. J. Agric. Food Chem. 2005; 53(22),
8688–8693.
Zhonghai L., Jie B., Jilie L., Haiyan Z. Analysis of volatile
components of three kumquat species by HS-SPME/GC-MS. J. Chinese
Cereals Oils Assoc. 2009; 09.
How to cite this article:
Nabaweya A. Ibrahim, Seham S. El-Hawary, Magdy M. D.
Mohammed, Mohamed A. Farid, Nayera A. M. Abdel-Wahed,
Mohamed A. Ali, Eman A. W. El-Abd. Chemical Composition,
Antiviral against avian Influenza (H5N1) Virus and Antimicrobial
activities of the Essential Oils of the Leaves and Fruits of
Fortunella margarita, Lour. Swingle, Growing in Egypt. J App
Pharm Sci, 2015; 5 (01): 006-012.
... Type of compounds The raw material origin Greece (Corfu) [31] Vietnam [17] Columbia [26,27] Korea [25] China [32] Thailand [33] USA (California) [28] Iran [34] Japan [35] Egypt [36] Taiwan [3] Italy (Sicily) [22] Tested species [31] Vietnam [17] Columbia [26,27] Korea [25] China [32] Thailand [33] USA (California) [28] Iran [34] Japan [35] Egypt [36] Taiwan [3] Italy (Sicily) [22] Tested species significantly higher in fruit peels than in fruit pulp; moreover, it is almost twice as high in unripe fruits than in those intended for harvesting. However, if the harvest date is postponed in time, the polyphenol content starts to increase again. ...
... Type of compounds The raw material origin Greece (Corfu) [31] Vietnam [17] Columbia [26,27] Korea [25] China [32] Thailand [33] USA (California) [28] Iran [34] Japan [35] Egypt [36] Taiwan [3] Italy (Sicily) [22] Tested species [31] Vietnam [17] Columbia [26,27] Korea [25] China [32] Thailand [33] USA (California) [28] Iran [34] Japan [35] Egypt [36] Taiwan [3] Italy (Sicily) [22] Tested species significantly higher in fruit peels than in fruit pulp; moreover, it is almost twice as high in unripe fruits than in those intended for harvesting. However, if the harvest date is postponed in time, the polyphenol content starts to increase again. ...
Article
The kumquat tree occurs in practically all countries of the subtropical zone. Kumquat fruits with a characteristic shape, size, colour and taste are the smallest of edible citrus and the only ones that can be eaten as a whole with the peel. Those fruits are rich in fiber, carotenoids, vitamins and minerals. The most important components are polyphenols showing antioxidant properties, in particular flavonoids of which the one characteristic is phloretin. The essential oils extracted from them are rich in numerous terpenoids, responsible for fragrance and aroma. An extensive literature review was carried out to collect data on the chemical composition of kumquat fruit and other selected parts of the plant. These data, depending on the variety, origin, part of the plant and method of obtaining were compiled and presented in the appropriate tables and figures. The structures of the most important components were also presented. Due to their nutritional properties and biological activity (antioxidant, antimicrobial, antitumor etc.) these components are potential sources of ingredients for foods, phytopharmaceuticals and cosmetics.
... Another study performed on essential oils of leaves and fruits of Fortunella margarita (Citrus margarita) against avian influenza A virus showed greater antiviral potential of fruit oil with 80% virus inhibition as compared to its leaves, due to the presence of "α-terpineol" in fruit. Similar effects were observed with volatile oils of Curcuma zedoaria and Hypericin perforatum attributed to the presence of "curcumenol" and "hypericin" respectively [133]. This could be a potent candidate in fight against SARS-CoV-2, if checked through properly designed in-vitro and in-vivo studies. ...
Article
Full-text available
Recently, the outbreak of severe acute respiratory syndrome cornoavirus-2 (SARS-CoV-2), causing coronavirus disease 2019 (COVID-19), has become a great perturbation all around the globe and has many devastating effects on every aspect of life. Apart from the oxygen therapy and extracorporeal membrane oxygenation, Remdesivir and Dexamethasone have been proven to be efficacious against COVID-19, along with various vaccine candidates and monoclonal antibody cocktail therapy for Regeneron. All of these are currently at different stages of clinical trials. People with weak immunity are more prone to a severe infection of SARS-CoV-2. Therefore, early and judicious nutritional supplementation along with pharmacological treatment and clinician collaborations are critical in restituting the current situation. Nutritional supplements help in acquiring strong immunity to prevent the progression of disease any further. Vitamin C, vitamin D, selenium, zinc and many other nutritional and dietary supplements inhibit the production of inflammatory cytokines during a viral infection and prevents several unwanted symptoms of infection. Many dietary components like citrus fruits, black elderberry, ginger, and probiotics have the ability to attack viral replication. These supplements can also tame the overriding immune system during coronavirus infection. Keeping in view these facts, nutritional and dietary supplements can be used along with other management modalities. These nutritional and dietary supplements are potential candidates to curb the convulsive unfolding of novel COVID-19, in combination with other standard treatment protocols. In this review, various search engines were used to exploit available literature in order to provide a comprehensive review on nutritional and dietary supplements with respect to the viral infections. It will also provide a brief overview on some of the clinical trials that are in progress to assess the role of nutritional supplements, either alone or in combination with other pharmacological drugs, in fight against COVID-19.
... Santolina insularis essential oil (12.7% of compound 4) presented high activity against herpes simplex type 1 and type 2, accompanied by low cytotoxicity (Logu et al. 2000). Fortunella margarita essential oil (10.3% of compound 14 in leaves and 5.5% in fruits) presented activity against avian influenza-A virus (Ibrahim et al. 2015). ...
Article
Full-text available
The emergence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in China and its spread worldwide has become one of the biggest health problem due to the lack of knowledge about an effective chemotherapy. Based on the current reality of the SARS-CoV-2 pandemic, this study aimed to make a review literature about potential anti-coronavirus natural compounds guided by an in silico study. In the first step, essential oils from native species found in the Brazilian herbal medicine market and Brazilian species that have already shown antiviral potential were used as source for the literature search and compounds selection. Among these compounds, 184 showed high antiviral potential against rhinovirus or picornavirus by quantitative structure–activity relationship analysis. (E)-α-atlantone; 14-hydroxy-α-muurolene; allo-aromadendrene epoxide; amorpha-4,9-dien-2-ol; aristochene; azulenol; germacrene A; guaia-6,9-diene; hedycaryol; humulene epoxide II; α-amorphene; α-cadinene; α-calacorene and α-muurolene showed by a molecular docking study the best result for four target proteins that are essential for SARS-CoV-2 lifecycle. In addition, other parameters obtained for the selected compounds indicated low toxicity and showed good probability to achieve cell permeability and be used as a drug. These results guided the second literature search which included other species in addition to native Brazilian plants. The majority presence of any of these compounds was reported for essential oils from 45 species. In view of the few studies relating essential oils and antiviral activity, this review is important for future assays against the new coronavirus.
... Many essential oils have been shown to possess antiviral activity against a variety of enveloped and nonenveloped viruses such as HSV, influenza, adenovirus type 3, Junin virus, poliovirus, norovirus, and coxsackie virus B1 (Table 2). (Ibrahim, El-Hawary et al. 2015) Carvacrol, p-cymene, γ-terpinene, and β-caryophyllene Murine norovirus (El Moussaoui, Sanchez et al. 2013) Thymol, carvacrol, p-cymene, β-caryophyllene, γ-terpinene, α-zingiberene, camphene, β-sesquiphellandrene, αrcurcumene, β-phellandrene, and β-bisabolene HSV-2 (Koch, Reichling et al. 2008) Thymol, p-cymene, γ-terpinene, and (E)-Cinnamaldehyde Influenza A virus subtype H1N1 (Setzer 2016) (Hayashi, Imanishi et al. 2007) Camphor, limonene, p-mentha-1, 8-diene, α-pinene, γterpinene , germacrene D, β-caryophyllene, calamine, leptospermone, flavesone, viridiflorene, isoleptospermone, geranial, menthol, menthone, and isomenthone HSV-1 and HSV-2 (Brand, Roa-Linares et al. 2016) (Reichling, Koch et al. 2005) (Schnitzler, Schuhmacher et al. 2008) (Schuhmacher, Reichling et al. 2003) Patchoulol, δ-guaieno; gurjunene-α, α-guaiene, aromadendrene, and β-patchoulene Influenza A (H2N2) virus (Wu, Ju et al. 2013) (Wu, Li et al. 2011) treatments to fight viral infections or their symptoms (drugs). In the latter case, they may be administered through oral, injection, skin, or ocular routes depending on the nature of the formulation and the intended target. ...
Article
Full-text available
The COVID-19 pandemic has resulted in unprecedented increases in sickness, death, economic disruption, and social disturbances globally. However, the virus (SARS-CoV-2) that caused this pandemic is only one of many viruses threatening public health. Consequently, it is important to have effective means of preventing viral transmission and reducing its devastating effects on human and animal health. Although many antivirals are already available, their efficacy is often limited because of factors such as poor solubility, low permeability, poor bioavailability, un-targeted release, adverse side effects, and antiviral resistance. Many of these problems can be overcome using advanced antiviral delivery systems constructed using nanotechnology principles. These delivery systems consist of antivirals loaded into nanoparticles, which may be fabricated from either synthetic or natural materials. Nevertheless, there is increasing empHasis on the development of antiviral delivery systems from natural substances, such as lipids, phospholipids, surfactants, proteins, and polysaccharides, due to health and environmental issues. The composition, morphology, dimensions, and interfacial characteristics of nanoparticles can be manipulated to improve the handling, stability, and potency of antivirals. This article outlines the major classes of antivirals, summarizes the challenges currently limiting their efficacy, and highlights how nanoparticles can be used to overcome these challenges. Recent studies on the application of antiviral nanoparticle-based delivery systems are reviewed and future directions are described.
... The essential oil constituents of Pogostemoncablin have been found active against the H2N2 influenza-A virus [119]. The anti-avian influenza virus H5N1 activity was reported by the essential oils from leaves and fruits of Fortune llamargarita [120]. The Trachyspermum oil has been reported of producing remarkable antiviral potential against the Japanese encephalitis virus (JEV). ...
Article
Essential oils and their chemical constituents have been reported with well documented antimicrobial effects against a range of bacterial, fungal and viral pathogens. By definition, essential oils are a complex mixture of volatile organic compounds which are synthesized naturally in different parts of the plant as part of plants secondary metabolism. The chemical composition of the essential oils is dominated by the presence of a range of compounds including phenolics, terpenoids, aldehydes, ketones, ethers, epoxides and many others inferring that essential oils must be effective against a wide range of pathogens. This review article mainly focuses on the antiviral potential of essential oils and their chemical constituents especially against influenza and coronaviruses. Essential oils have been screened against several pathogenic viruses, including influenza and other respiratory viral infections. The essential oils of cinnamon, bergamot, lemongrass, thyme, lavender have been reported to exert potent antiviral effects against influenza type A virus. The essential oil of Citrus reshni leaves has been shown to be effective against H5N1 virus. The essential oil of Lippia species at a concentration of 11.1 μg/mL has been shown to induce 100% inhibition of yellow fever virus in Vero cells. Essential oils and oleoresins have been shown through in vitro and in vivo experiments to induce antiviral effects against Coronavirus infectious bronchitis virus. A study reported 221 phytochemical compounds and essential oils to be effective against severe acute respiratory syndrome associated coronavirus (SARS-CoV) using a cell-based assay measuring SARS-CoV-induced cytopathogenic effect on Vero E6 cells. The main mechanism of antiviral effects of essential oils has been found to cause capsid disintegration and viral expansion which prevents the virus to infect host cells by adsorption via the capsid. Essential oils also inhibit hemagglutinin (an important membrane protein of various viruses) of certain viruses; this membrane protein allows the virus to enter the host cell. Many essential oils and their components could inhibit the late stages of viral life cycle by targeting the redox signalling pathway. Essential oils of Thymus vulgaris, cymbopogon citratus and Rosmarinus officinalis have been found to destabilize the Tat/TAR-RNA complex of HIV-1 virus, this complex being essential for HIV-1 replication. Being lipophilic in nature, essential oils can penetrate viral membranes easily leading to membrane disintegration. The current comprehensive review will facilitate researchers to find chemical entities from plant sources as possible inhibitory agents against various viruses.
... Essential oils as antibacterials, antifungals, antioxidants, etc., have been extensively investigated in numerous studies for decades [8][9][10]. Virucidal effects of EOs extracted from numerous aromatic and herbal plants are also well documented on a variety of viruses, such as IFV, HSV, HIV, yellow fever virus, and avian influenza, etc. [11][12][13][14]. In a recent study, Cagno et al. [2] reported that the EO from Salvia desoleana Atzei & V. Picci significantly suppressed the acyclovir resistant HSV-2 strains with a 50% effective concentration value (IC50) of 28.57 ...
Article
Full-text available
The presence of resistance to available antivirals calls for the development of novel therapeutic agents. Plant-derived essential oils may serve as alternative sources of virus-induced disease therapy. Previous studies have demonstrated essential oils to be excellent candidates to treat antiviral-resistant infection associated with their chemical complexity which confers broad-spectrum mechanisms of action and non-specific antiviral properties. However, almost no comprehensive reviews are updated to generalize knowledge in this regard and disclose the interplay between the components and their antiviral activities. This review provides an up-to-date overview of the antiviral efficacy of essential oils from a wide range of plant species and their characteristic components, as well as their overall mechanisms of action, focusing on the last decade. The roles of individual components relative to the overall antiviral efficacy of essential oils, together with the antiviral activity of essential oils in comparison with commercial drugs are also discussed. Lastly, the inadequacies in current research and future research are put forward. This review will provide references in the design of new drug prototypes and improve our understanding of the proper applications of essential oils in the future.
... Both samples were tested for antiviral activity against avian influenza (H5N1) virus, and the obtained results revealed that the fruit essential oil was more effective (80% virus inhibition by the MTT (3-(4,5-dimethylthiazol-2-yl)-2,5diphenyltetrazolium bromide) assay using Madin−Darby canine kidney (MDCK) cells for virus propagation). The IC50 values obtained for the leaf and fruit essential oils were 38.89 and 6.77 g/mL, respectively [53]. ...
Article
Full-text available
Essential oils have shown promise as antiviral agents against several pathogenic viruses. In this work we hypothesized that essential oil components may interact with key protein targets of the 2019 severe acute respiratory syndrome coronavirus 2 (SARS−CoV−2). A molecular docking analysis was carried out using 171 essential oil components with SARS−CoV−2 main protease (SARS−CoV−2 M pro), SARS−CoV−2 endoribonucleoase (SARS−CoV−2 Nsp15/NendoU), SARS−CoV−2 ADP−ribose−1″−phosphatase (SARS−CoV−2 ADRP), SARS−CoV−2 RNA−dependent RNA polymerase (SARS−CoV−2 RdRp), the binding domain of the SARS−CoV−2 spike protein (SARS−CoV−2 rS), and human angiotensin−converting enzyme (hACE2). The compound with the best normalized docking score to SARS−CoV−2 M pro was the sesquiterpene hydrocarbon (E)−β−farnesene. The best docking ligands for SARS−CoV Nsp15/NendoU were (E,E)−α−farnesene, (E)−β−farnesene, and (E,E)−farnesol. (E,E)−Farnesol showed the most exothermic docking to SARS−CoV−2 ADRP. Unfortunately, the docking energies of (E,E)−α−farnesene, (E)−β−farnesene, and (E,E)−farnesol with SARS−CoV−2 targets were relatively weak compared to docking energies with other proteins and are, therefore, unlikely to interact with the virus targets. However, essential oil components may act synergistically, essential oils may potentiate other antiviral agents, or they may provide some relief of COVID−19 symptoms.
Article
There is a need for new effective antivirals, particularly in response to the development of antiviral drug resistance and emerging RNA viruses such as SARS-CoV-2. Plants are a significant source of structurally diverse bioactive compounds for drug discovery suggesting that plant-derived natural products could be developed as antiviral agents. This article reviews the antiviral activity of plant-derived natural products against RNA viruses, with a focus on compounds targeting specific stages of the viral life cycle. A range of plant extracts and compounds have been identified with antiviral activity, often against multiple virus families suggesting they may be useful as broad-spectrum antiviral agents. The antiviral mechanism of action of many of these phytochemicals is not fully understood and there are limited studies and clinical trials demonstrating their efficacy and toxicity in vivo. Further research is needed to evaluate the therapeutic potential of plant-derived natural products as antiviral agents.
Article
Essential oils and isolated essential oil compounds are known to exert various pharmacological effects, such as antibacterial, antifungal, antiviral, anti-inflammatory, anti-immunomodulatory, antioxidant, and wound healing effects. Based on selected articles, this review deals with the potential antiviral and virucidal activities of essential oils and essential oil compounds together with their mechanism of action as well as in silico studies involving viral and host cell-specific target molecules that are indispensable for virus cell adsorption, penetration, and replication. The reported in vitro and in vivo studies highlight the baseline data about the latest findings of essential oils and essential oil compounds antiviral and virucidal effects on enveloped and non-enveloped viruses, taking into account available biochemical and molecular biological tests. The results of many in vitro studies revealed that several essential oils and essential oil compounds from different medicinal and aromatic plants are potent antiviral and virucidal agents that inhibit viral progeny by blocking different steps of the viral infection/replication cycle of DNA and RNA viruses in various host cell lines. Studies in mice infected with viruses causing respiratory diseases showed that different essential oils and essential oil compounds were able to prolong the life of infected animals, reduce virus titers in brain and lung tissues, and significantly inhibit the synthesis of proinflammatory cytokines and chemokines. In addition, some in vitro studies on hydrophilic nano-delivery systems encapsulating essential oils/essential oil compounds exhibited a promising way to improve the chemical stability and enhance the water solubility, bioavailabilty, and antiviral efficacy of essential oils and essential oil compounds.
Article
The human population worldwide has been challenged since December 2019 by the emerging COronaVIrus Disease 2019 (COVID-19), caused by SARS-CoV-2. The severity of the infection ranges from mild symptoms such as fever, cough and dizziness to severe illness and even death. Herbal medicines have been long used to attenuate infectious diseases due to their lower side effects, low cost and reduced potential to cause resistance. Observations from traditional medicine revealed that many plant-derived essential oils can be effective in the prevention and treatment of viral-induced respiratory tract infections. By this review, we aimed to call for screening essential oils for prevention or treatment of COVID-19. This review further highlights the possibility that the antiviral properties of essential oils enable them to be engaged in nutraceutical and pharmaceutical industries. Pharmaceutical industries are in need of ecofriendly effective antiviral drugs and essential oils may be the future core for lead drug bio-discovery.
Article
Full-text available
The aims of this study were to analyze the chemical composition of leaves essential oil of Warionia saharae, to evaluate the antioxidant capacity (DPPH test), antibacterial properties and to investigate toxicity against Artemia salina of both oil and ethanolic extract. The results showed that essential oil was toxic (CL50 = 1,56 μg/ml). The GC–MS analysis revealed 52 compounds representing 91.54% of the total oil containing Nerolidol (25,95%) and β-Eudesmol (38,12%) as a major components. The oil (1 mg/ml) exhibited a strong antibacterial effect as a diameter of zones of inhibition (28,5 ± 2,12 and 37,5 ± 3.53 mm) against St. aureus and P. aeruginosa, respectively. While, ethanolic extract (50 mg/ml) exhibited a moderate effect against all tested bacterial strains. MICs values of oil and the extract were ranged 0,031-0,25 μg/ml and 6,25-12,5 mg/ml, respectively. The free radical scavenging activities of the oil was higher than that of ethanolic extract (IC50 = 21,49 and IC50 = 182 μg/ml, respectively).
Article
Full-text available
The aim of this study was to determine the main constituents of the essential oil isolated from Fortunella crassifolia Swingle peel by hydro-distillation, and to test the efficacy of the essential oil on antimicrobial activity. Twenty-five components, representing 92.36% of the total oil, were identified by GC-MS analysis. The essential oil showed potent antimicrobial activity against both Gram-negative (E. coli and S. typhimurium) and Gram-positive (S. aureus, B. cereus, B. subtilis, L. bulgaricus, and B. laterosporus) bacteria, together with a remarkable antifungal activity against C. albicans. In a food model of beef extract, the essential oil was observed to possess an effective capacity to control the total counts of viable bacteria. Furthermore, the essential oil showed strongly detrimental effects on the growth and morphological structure of the tested bacteria. It was suggested that the essential oil from Fortunella crassifolia Swingle peel might be used as a natural food preservative against bacteria or fungus in the food industry.
Article
Full-text available
Crude extracts of leaves and bark of E. jambolana were tested for antiviral activity against highly pathogenic avian influenza virus (H5N1) by CPE reduction assay in three different layouts to elucidate virucidal, post-exposure and preexposure antiviral activity of the extracts. The cold and hot aqueous extracts of bark and hot aqueous extract of leaves of E. jambolana showed significant virucidal activity (100% inhibition) which was further confirmed in virus yield reduction assay (-98 to 99% reduction) and by egg based in ovo assay. The selective index (CC50/EC50) of hot aqueous extract (248) and cold aqueous extract (43.5) of bark of E. jambolana showed their antiviral potential against H5N1 virus. The significant virucidal activity of leaves and bark of E. jambolana merits further investigation as it may provide alternative antiviral agent for managing avian influenza infections in poultry farms and potential avian-human transmission.
Article
Aim: To investigate the antiviral effect of compound of Curcuma zedoaria volatile oil and Hypericin perforatum extract liquid on H5N1 avian influenza virus (AIV) in MDCK cell line and non-AIV-immunized chickens. Methods: CPE inhibiton and MTT assay were used to study the inhibition of compound of Curcuma zedoaria volatile oil and Hypericin perforatum extract liquid, curcumenol and hypericin on H5N1 in MDCK, the retarding effect on H5N1 infection, the inhibition on the virus adsorption, and the therapeutic effect on the virus-infected cells. In addition, after one-week oral feeding of the liquid, non-AIV-immunized chickens were exposed to H5N1 AIV and the protecting effects of the liquid on the chickens were evaluated. The effect was also assessed after the chickens were simultaneously exposed to the liquid and and the attacking H5N1. The antiviral effects of oseltamivir and amantadine were also detected as the control groups. Results: In vitro results showed that compound of Curcuma zedoaria volatile oil and Hypericin perforatum extract liquid was effective via the mechanisms including virus killing, virus inhibiting, the prevention of the virus spread, and the treatment of the virus-related injury in MDCK. In vivo studies indicated that compound of Curcuma zedoaria volatile oil and Hypericin perforatum extract liquid prolonged the mean survival days of the chickens, showing significant preventive effect in the chickens. It was also proved that compound of Curcuma zedoaria volatile oil and Hypericin perforatum extract liquid was more effective than zedoary oil and Hypericum perforatum extract. Conclusion: Compound of Curcuma zedoaria volatile oil and Hypericin perforatum extract liquid is superior to zedoary oil and Hypericum perforatum in the treatment of H5N1 via the prevention of the virus infection and it might be very promising in the development into into effective anti-H5N1 virus drug.
Article
Recent work in the field of biologically active, essential oils is reviewed. Essential oil extraction methods that are covered include cold pressing, extraction with other essential oils, steam distillation, solvent extraction, supercritical fluid extraction, and solid phase extraction. Separation methods for the isolation of individual constituents that are covered include GC, LC, and distillation. Biological activities of essential oils and their components, including antiallergic, enzyme inhibitory, psychological, anti-inflammatory, antimutagenic, anticarcinogenic, antiviral, insect repellent, molluscicidal, and antimicrobial are also reviewed. In particular, several examples of our own and others' work in this area that are discussed include, 1) the structure and antimutagenic activity of new sesquiterpenoid eudesmol derivatives, 2) the biological activity and odor perception of optically active rose oxides, 3) the polyphenol oxidase inhibitory activity of acyclic terpene alcohols, commonly found in essential oils, that are used in cosmetic applications, 4) the effects of the diterpene phenol, totarol, in combination with known antibiotics, on a methicillin resistant Staphylococcus aureus(MRSA) strain, and 5) the synergistic antimicrobial activity of the combination of perillaldehyde and polygodial, constituents of two herbs found in traditional Japanese foods. Also reviewed are the various uses and applications of essential oils that have been reported recently; examples included in the discussion are insect and animal repellents, oral care products, pharmaceuticals, drug delivery systems, topical applications, cosmetics, food preservatives, antioxidants, industrial applications, and the uses in packaging and consumer household products.
Article
In this study 120 compounds were found and 71 volatile compounds were identified in the essential oil of kumquat. A simultaneous distillation/extraction technique was used to obtain the essential oil from the fruit. Individual compounds were identified by means of Kovats retention indices on polar and nonpolar columns and mass spectral data. Some 13 sesquiterpenes, 8 terpenes, 11 alcohols, 1 ketone, 8 aldehydes and 13 esters were identified in kumquat oil for the fist time. Limonene was the most abundant compound comprising 93% of the whole oil.
Article
The relative antiviral activities of four drugs against contemporary strains of influenza A and B viruses were determined in Madin-Darby canine kidney cell monolayers with a plaque inhibition assay. This assay proved to be a reliable, rapid method of determining 50% inhibitory concentrations that correlated well with clinically achievable drug levels and the results of clinical trials. Contemporary strains of influenza A viruses (subtypes H1N1, H3N2, HSW1N1) required amantadine hydrochloride and rimantadine hydrochloride 50% inhibitory concentrations in the range of 0.2 to 0.4 microgram/ml, whereas 50% inhibitory concentrations ranged from approximately 50 to 100 micrograms/ml against influenza B viruses. Ribavirin was approximately 10-fold less active than amantadine hydrochloride against influenza A viruses, and the ribavirin 50% inhibitory concentrations against both influenza A and B viruses ranged from 2.6 to 6.8 micrograms/ml. Inosiplex had no antiviral activity in this test system.
Article
The antifungal activity of essential oil isolated from the leaves of bael (Aegle marmelos (L.) Correa ex Roxb., Rutaceae) has been evaluated using spore germination assay. The oil exhibited variable efficacy against different fungal isolates and 100% inhibition of spore germination of all the fungi tested was observed at 500 ppm. However, the most resistant fungus, Fusarium udum was inhibited 80% at 400 ppm. Kinetic studies showed concentration as well as time dependent complex inhibition of spore germination by the essential oil.