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Chemical composition and antioxidant, antibacterial and
antifungal activities of the essential oils from
Bidens pilosa Linn. var. Radiata
Farah Deba, Tran Dang Xuan, Masaaki Yasuda, Shinkichi Tawata
*
Department of Bioscience and Biotechnology, Faculty of Agriculture, University of the Ryukyus, 1 Senbaru, Nishihara-cho, Okinawa 903-0213, Japan
Received 17 January 2007; received in revised form 12 April 2007; accepted 17 April 2007
Abstract
The present study describes the chemical composition, antibacterial and antifungal activities of essential oils from Bidens pilosa,a
traditional medicinal plant widely distributed in the subtropics and tropics. The essential oils from the fresh leaves and flowers of B.
pilosa were analyzed by GC–MS. Forty-four components were identified, of which b-caryophyllene (10.9% and 5.1%) and s-cadinene
(7.82% and 6.13%) were the main compounds in leaves and flowers, respectively. The oils and aqueous extracts of leaves and flowers
were subjected to screening for their possible antioxidant activities by using 2,20-diphenyl-1-picrylhydrazyl (DPPH) and b-carotene
bleaching methods. In the former case, the essential oils from leaves and flowers were found to be superior to all aqueous extracts tested
with an IC
50
value of 47.5 and 49.7 lg/ml, respectively, whereas all extracts and essential oils seemed to inhibit the oxidation of linoleic
acid in the latter case. The oils from B. pilosa exerted significant antibacterial and antifungal activities against six bacteria and three fun-
gal strains. The inhibitory activity of the flower essential oils in Gram-negative bacteria was significantly higher than in Gram-positive.
Our findings demonstrate that the essential oils and aqueous extracts of B. pilosa possess antioxidant and antimicrobial activities that
might be a natural potential source of preservative used in food and other allied industries.
Ó2007 Elsevier Ltd. All rights reserved.
Keywords: Essential oils; Antifungal activity; Bidens pilosa; Antibacterial activity
1. Introduction
The food industry at present is facing a tremendous pres-
sure from consumers for using chemical preservatives to pre-
vent the growth of food borne and spoiling microbes. To
reduce or eliminate chemically synthesized additives from
foods is a current demand worldwide. A new approach to
prevent the proliferation of microorganism or protect food
from oxidation is the use of essential oils as preservatives.
Essential oils of plants are of growing interest both in the
industry and scientific research because of their antibacte-
rial, antifungal, and antioxidant properties and make them
useful as natural additives in foods (Pattnaik, Subraman-
yam, Bapaji, & Kole, 1997). Free radical oxidation of the
lipid components in food due to the chain reaction of lipid
peroxidation is a major strategic problem for food manufac-
turers. Due to undesirable influences of oxidized lipids on
the human organisms, it is essential to decrease lipid perox-
idation products in food (Karpin
´ska, Borowski, & Dan-
owska-Oziewicz, 2001). Reactive oxygen species are
reported to be a causative agent of various diseases such as
arthritis, asthma, dementia, mongolism, carcinoma and Par-
kinson’s disease (Perry et al., 2000). Plant essential oils and
their extracts have had a great usage in folk medicine, food
flavoring, fragrance, and pharmaceutical industries (Kus-
menoglu, Baser, & Ozek, 1995).
Bidens pilosa Linn. var. Radiata (family Asteraceae) is
widely distributed in the subtropical and tropical regions
of the world. It is 30–100 cm in height with yellow flowers
and is well known as hairy beggar ticks, sticks tights, and
Spanish needles. The plant is used in various folk medicines
0956-7135/$ - see front matter Ó2007 Elsevier Ltd. All rights reserved.
doi:10.1016/j.foodcont.2007.04.011
*
Corresponding author. Tel.: +81 98 895 8803; fax: +81 98 895 8734.
E-mail address: b986097@agr.u-ryukyu.ac.jp (S. Tawata).
www.elsevier.com/locate/foodcont
Available online at www.sciencedirect.com
Food Control 19 (2008) 346–352
such as anti-inflammatory, antiseptic, liver-protective,
blood-pressure lowering, and hypoglycemic effects (Dimo
et al., 2002). Phenylpropanoid glucosides, polyacetylenes,
a diterpenes, flavonoids, and flavone glycosides have been
identified as bioactive components from this plant (Chiang
et al., 2004). These compounds were suggested to be
involved in the antioxidant (Chiang et al., 2004), antibacte-
rial and antimicrobial activities (Rabe & Staden, 1997). The
plant has been widely used in Taiwan as a traditional medi-
cine and as a major ingredient of herbal tea, which is believed
to prevent inflammation and cancer (Yang et al., 2006).
To the best of our knowledge, there are no available
reports on chemical composition and biological activities
of the essential oils from B. pilosa. Therefore, the aim of
the present work was carried out to study in vitro antioxi-
dant, antibacterial, and antifungal activities of the essential
oils as well as the water extract of B. pilosa in addition to
evaluate the component of essential oils by GC–MS. Essen-
tial oils obtained by steam-distillation were analyzed for
their possible antioxidant activities by two complementary
methods, namely DPPH free radical-scavenging and b-car-
otene bleaching methods due to identify all possible mech-
anisms characterizing an antioxidant.
2. Materials and methods
2.1. Plant material, extraction of essential oils, and
preparation of the water extract
The fresh leaves and flowers of B. pilosa at vegetable
stage were collected from nature around the campus of
University of the Ryukyus, Okinawa, Japan, in April
2006. Fresh leaves or flowers (each 400 g) of B. pilosa were
steam-distilled for 4 h. The distillates of each were
extracted with 200 ml diethyl ether and conducted twice.
The solvent was carefully removed under vacuum at
30 °C. The essential oils thus obtained were stored at
4°C for testing and analyzing. After completion of
steam-distillation, the aqueous mixtures of leaves or flow-
ers were filtered to collect water extract which was evapo-
rated to dryness on a rotary evaporator at 40 °C.
2.2. Identification by GC–MS
An aliquot of 1 ll oils dissolved in diethyl ether and
adjusted to 1000 ppm was injected into GC–MS (QP-
2010, Shimadzu Co., Kyoto, Japan). The DB-5MS column
was 30 m in length, 0.25 mm i.d., and 0.25 lm in thickness
(Agilent Technologies, J&W Scientific Products, Folsom,
CA). The carrier gas was helium. The operating condition
of GC oven temperature was maintained as: initial temper-
ature 50 °C for 5 min, programmed rate 5 °C/min up to
final temperature 280 °C with isotherm for 5 min. The
injector and detector temperatures were set at 250 and
280 °C, respectively. The essential oil components were
identified by comparing their retention times and mass
fragmentation pattern with those of standards and MS
library (Shimadzu’s GCMS solution software, version
2.4). The quantity of oil components was compared using
peak area measurements.
2.3. Antioxidant activity
2.3.1. DPPH assay
In this assay, antioxidant activity of essential oils was
evaluated by measuring the bleaching of the purple-colored
ethanol solution of DPPH (Burits & Bucar, 2000). The rad-
ical scavenging ability was determined according to the
method described by Abe, Murata, and Hirota (1998).
One milliliters from a 0.5 mM ethanol solution of the
DPPH radical was mixed to 2.0 ml of different concentra-
tions of essential oils from leaves and flowers and was
added 2 ml of 0.1 M sodium acetate buffer (pH 5.5). The
mixtures were well shaken and kept at room temperature
in the dark for 30 min. The absorbance was measured at
517 nm using a Shimadzu UV–Vis spectrophotometer mini
1240, Kyoto (Japan). The authentic a-tocopherol and butyl
hydroxyl toluene (BHT) were used as a positive control
while ethanol was as negative one. Inhibition (I%) of
DPPH radical was calculated using the equation:
I%¼ðAo As=AoÞ100
Where Ao is the absorbance of the control (containing all re-
agents except the test compound), and As is the absorbance
of the test compound. The IC
50
value represented the con-
centration of the essential oils that caused 50% inhibition.
2.3.2. b-Carotene bleaching assay
In this assay, antioxidant capacity is determined by mea-
suring the inhibition of the volatile organic compounds and
the conjugated diene hydroperoxides arising from linoleic
acid oxidation (Dapkevicius, Venskutonis, Van Beek, &
Linseen, 1998). Antioxidant activity was carried out
according to the b-carotene bleaching method (Siddhuraju
& Becker, 2003) with minor modifications. b-Carotene
(2.0) mg was dissolved in 10 ml chloroform. Linoleic acid
(20 ll) and Tween-40 (200 mg) were mixed with 1 ml of
the chloroform solution. The chloroform was evaporated
under vacuum at 45 °C, then 50 ml oxygenated water was
added, and the mixture was vigorously shaken. The emul-
sion obtained was freshly prepared before each experiment.
The b-carotene-linoleic acid emulsion (250 ll) was trans-
ferred to a 96-well microplate. Ethanolic solutions (30 ll)
of the sample extracts, BHT and a-tocopherol at
1000 ppm were added onto plate. An equal amount of eth-
anol was used as control. Absorbance was taken at 492 nm
after incubation for every 15 min until 180 min at 45 °C
using a microplate reader (Benchmark plus microplate
170-6930j1, BIO-RAD Company).
2.4. Antibacterial activities
All bacterial strains have been kindly provided by Lab-
oratory of Microbiology, University of the Ryukyus, Oki-
F. Deba et al. / Food Control 19 (2008) 346–352 347
nawa, Japan. Antibacterial activities of all extracts using
ampicillin as the reference standard were tested against
Bacillus subtilis,Bacillus cereus,Bacillus pumilus,Esche-
richia coli and Pseudomonus ovalis. Essential oils, water
extracts and ampicillin were dissolved in 10% Tween 80
which was used as the control. Antibacterial activity was
determined by using the disc diffusion method (Karaman
et al., 2003). One hundred microliters of test organisms
[10
6
colony forming units CFU/ml] grown in nutrient broth
media for 24 h were spread over the surface of solid nutri-
ent agar medium in 9 cm diameter Petri dishes. Filter paper
discs (6 mm in diameter) loaded with each of the samples
and ampicillin were placed on the surface of the nutrient
agar which were incubated at 37 °C for 24 h, and then
the diameters of inhibition zones were measured in
millimeters.
2.5. Fungicidal bioassay
The tested fungi including Corticum rolfsii f.sp. (Curzi,
Roma N.S.), Fusarium solani (MAFF 237472) and Fusar-
ium oxysporum (Mormodiace, S.K. Sun & J.W. Huang)
were kindly provided from the Laboratory of Plant Pathol-
ogy, University of the Ryukyus, Okinawa, Japan. C. rolfsii,
F. solani and F. oxysporum were isolated from Mangifera
indica,Nicotiana tabacum L. and Balsam pear plants,
respectively. The three phytopathogens have been charac-
terized by causing the following symptoms: basal rot dis-
ease of tomato, brown root rot in peanuts and fusarium
yellows on asparagus. These phytopathogens were cultured
on potato dextrose agar (PDA) (1.5% agar) for 4 days at
27 °C before the experiment was carried out. One ml vol-
ume of each diluted solution was mixed with 8.0 ml of
PDA and was then put in the Petri dish. Finally, the spores
from a mycelia disk (5.7 mm in diameter) were put on the
centre of the PDA plate. All plates were put into an incu-
bator (27 °C) for 4 days and controls contained 10% Tween
80 only. The antifungal activity was measured by agar dilu-
tion method (Taira, Tawata, Kobamoto, Toyama, & Yas-
uda, 1994) and expressed as percentage inhibition against
the mycelia growth diameter.
2.6. Statistical analysis
Means and standard errors (SE) of the samples were cal-
culated. Each treatment was carried out with three repli-
cates. Mean differences were determined by using Fisher’s
Protected LSD test at 5% level of significance. All statisti-
cal analyses were performed using SAS version 8.2 (SAS
Institute, 1999–2001).
3. Results and discussion
3.1. Chemical composition of the essential oils
Fresh leaves and flowers part of B. pilosa were subjected
to steam-distillation and the colorless and yellowish essen-
tial oils were obtained 0.08% and 0.06%, w/w, respectively.
The results obtained by GC–MS analysis of the essential oils
from B. pilosa are presented in Table 1. Forty-four compo-
nents were identified and the major essential oils belonging
to terpenes were b-caryophyllene (10.9% and 5.1%) and s-
cadinene (7.82% and 6.13%), in the leaves and flowers,
respectively. The other chemical components were a-pinene,
limonene, b-trans-ocimene, b-cis-ocimene, s-muurolene, b-
bourbonene, b-elemene, b-cubebene, a-caryophyllene,
caryophyllene oxide, and megastigmatrienone.
To the best of our knowledge, the essential oils of B. pil-
osa have never been reported. This work is therefore the
Table 1
Essential oil components of fresh leaves and flowers of B. pilosa
Components RI
a
Peak area %
Leaves Flowers
Acetal 815 0.03 0.09
Cis-3-Hexen-1-ol 854 0.10 –
b
a-Pinene 930 0.99 5.97
Camphene 947 0.02 0.06
b-Phellandrene 971 0.01 0.15
b-Pinene 975 0.07 0.39
b-Myrcene 988 0.29 1.54
Cis-3-Hexenyl acetate 1005 0.77 –
3-Carene 1006 – 0.65
m-Cymol 1024 0.08 0.11
Limonene 1029 0.34 2.12
b-trans-Ocimene 1036 0.55 1.64
b-cis-Ocimene 1047 1.45 1.46
c-Terpinene 1059 – 0.05
b-Linalool 1100 0.43 0.09
(4E,6Z)-2,6-Dimethyl-2,4,6-octatriene 1129 0.24 0.70
trans-Verbenol 1144 – 0.11
cis-Verbenol 1148 0.11 0.18
4-Terpineol 1183 0.15 0.41
p-Cymen-8-ol 1198 0.26 0.35
Bornyl acetate 1287 0.15 0.18
Elixene 1313 0.25 0.32
a-Cubebene 1318 0.17 0.21
Ylangene 1326 0.13 0.13
s-Muurolene 1329 1.04 0.78
b-Bourbonene 1332 1.10 0.99
b-Elemene 1333 1.00 0.67
a-Bergamotene 1341 0.12 0.07
b-Caryophyllene 1345 10.9 5.1
b-Cubebene 1348 2.23 1.77
b-Farnesene 1353 0.72 0.29
a-Caryophyllene 1357 1.55 1.00
(+)-Epi-bicyclosesquiphellandrene 1358 0.27 0.25
Isoledene 1362 0.67 0.47
s-Cadinene 1365 7.82 6.13
b-Bisabolene 1372 0.31 0.03
b-Gurjunene 1375 0.44 0.24
()-b-Cadiene 1376 0.82 0.23
2,5,9-Trimethylcycloundeca-4,8-dienone 1387 0.82 0.16
trans-Nerolidol 1388 0.39 0.13
Ent-spathulenol 1396 0.33 0.18
Caryophyllene oxide 1398 1.47 1.03
Megastigmatrienone 1444 5.35 2.04
Diphenylenemethane 1450 1.94 1.77
a
Retention index relative to n-alkanes on the DB-5 column.
b
Not detected.
348 F. Deba et al. / Food Control 19 (2008) 346–352
first report on the essential oils from these tropical and sub-
tropical species. Among identified compound, b-caryophyl-
lene is well-known for its anti-inflammatory and local
anaesthetic activities (Ghelardini, Galeotti, Mannellia,
Mazzanti, & Bartolini, 2001). This compound is also used
in spice blends, soaps, detergents, creams and lotions,
and is widely used in food products and beverages (Sko
¨ld,
Karlberg, Matura, & Bo
¨rje, 2006).
3.2. Antioxidant activity
3.2.1. DPPH free radical scavenging activity
Free radical scavenging capacities of the extracts, mea-
sured by DPPH assay are shown in Fig. 1. There is no sig-
nificant difference between leaves and flower essential oils.
The essential oils of both plant parts had stronger activity
than the aqueous extracts. The activity of the extracts is
proportional to the concentrations and the lower IC
50
value reflects better protective action. The leaves and flow-
ers essential oils were able to reduce the stable free radical
2,20-diphenyl-1-picrylhydrazyl (DPPH) to the yellow col-
ored diphenylpicrylhydrazine with an IC
50
of 47 and
50 lg/ml, whereas those of the synthetic and natural anti-
oxidant activity were 21 and 36 lg/ml, respectively. The
flower aqueous extracts were found to be less efficient in
radical scavenging with an IC
50
value of 172 lg/ml,
whereas leaves aqueous extracts was 61 lg/ml. B. pilosa
essential oils contained monoterpenes and oxygenated terp-
enes such as a-pinene, ocimene, verbenol and b-myrcene.
Moreover, trying to correlate the observed activity with
the chemical composition of the oils, it is noteworthy to
cite the work of Ruberto and Baratta (2000), who studied
the antioxidant activity of 98 pure essential oils chemical
components and showed that monoterpene hydrocarbons
had a significant protective effect, with several variants
due to the different functional groups. Furthermore, some
researchers show that some essential oils rich in non-
phenolic compounds also have antioxidant potentials
(El-Massry, El-Ghorab, & Farouk, 2002). Table 1 shows
that essential oils of B. pilosa are markedly rich in non-
phenolic components. Because of this, activities of B. pilosa
essential oils can be attributed to the nonphenolic
constituents.
3.2.2. Antioxidant activity measured by b-carotene
bleaching method
In the b-carotene bleaching method, the degree of lino-
leic acid oxidation is determined by measuring oxidation
products (lipid hydroperoxides, conjugated dienes, and vol-
atile by-products) of linoleic acid which simultaneously
attack b-carotene, resulting in bleaching of its characteris-
tic yellow color in ethanolic solution. All extracts of B.pil-
osa inhibit the oxidation of linoleic acid, which is an
important issue in food processing and preservation
(Fig. 2). Inhibition of the breakdown of lipid hydroperox-
ides to unwanted volatile products allows us to determine
secondary antioxidants in related mechanisms. It has been
well reported that phenolic compounds are able to donate a
hydrogen atom to the free radicals thus stopping the prop-
agation chain reaction during lipid oxidation process (Sa
´n-
chez-Moreno, Larrauri, & Sauro-Calixto, 1998). Leaves
essential oils and aqueous extracts of leaves and flowers
exhibited the higher antioxidant activities than those of
the flower oils. The lower activity of the flower essential oils
of B. pilosa may also be due to its volatility at higher tem-
perature. The antioxidant effect of essential oils depends
not only on the temperature but also on many factors such
as their structural features, the character of the lipid sys-
tem, and on the binding of the fatty acids.
3.3. Antibacterial activities
As can be seen in Table 2, essential oils and the water
extracts obtained from B. pilosa showed moderate antimi-
crobial activity against all microorganisms tested. In gen-
eral, leaves essential oils exhibited higher stronger activity
than the others. The antibacterial activity of essential oils
and aqueous extracts were in the range of less than 7 mm
and not higher than 20 mm. Among all essential oils and
extracts, the flower essential oils were the most active
against E. coli inhibition. It has been demonstrated that
monoterpene hydrocarbons and oxygenated monoterpenes
in the flower essential oils are able to destroy cellular integ-
rity, and thereby inhibit respiration and ion transport pro-
cesses. This is strongly supported by the study on the effects
of different essential oils components on outer membrane
permeability in Gram-negative bacteria (Helander et al.,
1998). The sesquiterpene b-caryophyllene is known to play
a critical part in plant defense (Ulubelen et al., 1994). The
maximum activity of leaves essential oils was observed
against Gram-positive bacteria B. cereus and B. subtilis
but these oils had poor activity on the growth of Gram-
negative bacteria. Most studies investigating on the action
of essential oils against food spoilage organisms and food
borne pathogens agree that, essential oils are slightly more
active against Gram-positive than Gram-negative bacteria
(Lambert, Skandamis, Coote, & Nychas, 2001). The anti-
bacterial activity of the essential oils of B. pilosa against
0
20
40
60
80
100
120
140
160
180
Leaves oil
Flower oil
Leaves
extract
Flower
extract
α−
tocopherol
BHT
Plant
p
arts
DPPH IC
50
(µg/ml)
ccb
a
de
Fig. 1. Free radical scavenging activities of the essential oil and aqueous
extracts from B. pilosa. BHT and a-tocopherol as a positive controls.
Values are means of three replications ± SE.
F. Deba et al. / Food Control 19 (2008) 346–352 349
studied bacteria may be due to the presence of a high con-
centration of b-caryophyllene, since the antimicrobial
properties of caryophyllene and caryophyllene oxide found
in Salvia sclarea were also observed by Ulubelen et al.
(1994). The antimicrobial activity of the essential oils of
B. pilosa is apparently related to its terpenes type compo-
nents such as pine, myrcene, limonene, ocimene, linalool
and verbenol (Table 1), since there is a relationship
between the chemical structures of the most abundant oils
and their antimicrobial activities. Although the mechanism
of action of terpenes is not fully understood, it is thought
to involve membrane disruption by the lipophilic com-
pounds (Cowan, 1999). The essential oils containing terp-
enes are also reported to possess antimicrobial activity
(Dorman & Deans, 2000), which are consistent with our
present studies.
3.4. Antifungal activities
The results of antifungal activity assays showed that the
essential oils and aqueous extract of B. pilosa had inhibi-
tory effects on the growth of fungi (Table 3). F. solani
was most suppressed as its growth was mostly reduced by
all tested doses, followed by F. oxysporum and C. rolfsii.
On the other hand, the fungitoxic activities of the flower
essential oils were higher than those of the leaves. At
100 ppm, the essential oils of B. pilosa appeared to be effec-
tive against growth of these phytopathogens above 80%.
The volatile oils consist of complex mixtures of numerous
components. The major or trace compound(s) might give
rise to the antifungal activity. Possible synergistic and
antagonistic effects of compounds also play an important
role in fungi inhibition. Previous papers on the antifungal
activities of essential oils of some species of various genera
have shown that they have varying degrees of growth inhi-
bition effects against some agricultural pathogenic fungal
species (Alvarez-Castellanos, Bishop, & Pascual-Villalo-
bos, 2001). Although the different compounds exhibited
varying degrees of antifungal activity, b-caryophyllene
and caryophyllene oxide were very fungitoxic against the
studied Fusarium species (Cakir, Kordali, Zengin, Izumi,
& Hirata, 2004).
The present study indicates that b-caryophyllene and
caryophyllene oxide detected in B. pilosa essential oils
0
0.05
0.1
0.15
0.2
0.25
0.3
0.35
0.4
0 15 30 45 60 75 90 105 120 135 150 165 180 195
Time (min)
Absorbance at 492 nm
Control L. oil F. oil
L. aqueous extract F. aqueous extract α -tocopherol
BHT
Fig. 2. Antioxidant activities of essential oil and aqueous extracts from B. pilosa, BHT and a-tocopherol measured by b-carotene bleaching method. L,
leaves; F, flower.
Table 2
Antibacterial activities of the essential oils and extracts of Bidens pilosa
Test bacteria Essential oils and aqueous extracts from leaves and flowers
a
Zone of inhibition (mm)
b
L.
c
essential oil F.
d
essential oil L. extracts F. extracts Ampicilline
e
Control LSD (0.05)
Micrococcus flavus 12.7 ± 0.3 b 8.7 ± 0.3 d 10.2 ± 0.2 c 10.8 ± 0.3 c 44.3 ± 0.2 a 0 0.9
Bacillus subtilis 17.3 ± 1.9 b 11.7 ± 0.2 c 10.9 ± 0.1 c 10.3 ± 0.2 c 28.3 ± 1.7 a 0 3.6
Bacillus cereus 19.0 ± 1.4 a 11.2 ± 0.3 b 11.8 ± 0.4 b 18.5 ± 1.0 a 17.7 ± 1.3 a 0 3.2
Bacillus pumilus 12.3 ± 0.7 b 10.8 ± 0.2 c 10.5 ± 0.4 c 7.7 ± 0.2 d 29.7 ± 0.3 a 0 1.3
Escherichia coli 13.7 ± 0.4 c 20.3 ± 0.7 b 10.2 ± 1.1 d 14.0 ± 1.3 c 33.7 ± 0.2 a 0 2.5
Pseudomonus ovalis 12.5 ± 0.8 ab 13.7 ± 1.5 a 10.2 ± 0.6 b 12.5 ± 0.6 ab 15.3 ± 0.3 a 0 3.0
Values are means of three replications ± SE. Means with the same letter are not significantly different at P60.05.
a
400 lg/disc.
b
Diameter of disc 6 mm.
c
Leaves.
d
Flowers.
e
30 lg/disc.
350 F. Deba et al. / Food Control 19 (2008) 346–352
may play an important role in antifungal activities. a-
Pinene, which was found in appreciable amounts in the oils
of B. pilosa, has been reported to be the cause of the anti-
fungal activity of oils from Pistacia lentiscus (Magiatis,
Melliou, Skaltsounis, Chinou, & Mitaku, 1999). Another
minor monoterpene alcohol, linalool, is reported to have
a wide range of antibacterial and antifungal activity (Patt-
naik et al., 1997).
The essential oils from this plant exhibited antibacterial,
antifungal and antioxidant activities. The antioxidant
activity may be ascribed to the presence of the same chem-
ical components. Monoterpenes found in these essential
oils may act as radical scavenging agents. It seems to be
a general trend that the essential oils which contain mono-
terpene hydrocarbons, oxygenated monoterpenes and/or
sesquiterpenes have greater antioxidative properties (Tepe
et al., 2004). These activities may be attributed to the pres-
ence of p-cymene-8-ol, a-pinene, b-pinene, ocimene, limo-
nene, terpinene, and camphene found in B. pilosa
essential oils. Enatiomers of a-pinene, 2-b-pinene and lim-
onene have a strong antibacterial activity (Magiatis et al.,
1999; So
¨kmen et al., 2003). These chemical components
exert their toxic effects against studied microorganisms
through the disruption of bacteria or fungal membrane
integrity (Knobloch, Pauli, Iberal, Weis, & Weigand,
1989). The possible mechanisms of other essential oils com-
ponents, such as trans-caryophyllene, camphene, and a-
humulene have not yet been elucidated. The mechanism
of antifungal activity of these essential oils is still unknown.
Antioxidants, antibacterial and antifungal properties of
the essential oils and various extracts from many plants
have recently been of great interest in both research and
the food industry, because their possible use as natural
additives emerged from a growing tendency to replace syn-
thetic antioxidants with natural ones. Owing to strong anti-
fungal and protective features exhibited in antioxidant
activity tests, the essential oils and aqueous extracts of B.
pilosa could be considered a natural herbal source that
can be freely used in the food and pharmaceutical indus-
tries. This is the first report on the essential oils composi-
tion, antioxidants, antibacterial and antifungal activity of
B. pilosa. Further studies are needed to obtain more infor-
mation regarding the practical effectiveness of these oils in
animal models.
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Table 3
Antifungal activities of essential oils and aqueous extracts of B. pilosa
Plant parts Concentrations (ppm) Antifungal activities (% of inhibition)
Corticum rolfsii Fusarium solani Fusarium oxysporum
Leaves essential oil 100 85.7 ± 0.9 c 68.2 ± 0.0 d 74.9 ± 1.7 d
Leaves essential oil 250 96.0 ± 0.8 a 77.9 ± 1.8 c 87.9 ± 0.4 b
Flower essential oil 100 60.4 ± 0.9 e 89.2 ± 0.4 b 86.9 ± 0.5 b
Flower essential oil 250 89.4 ± 1.2 b 98.0 ± 0.3 a 94.9 ± 0.6 a
Leaves aqueous extract 100 44.6 ± 1.7 f 60.5 ± 2.1 e 71.6 ± 0.7 d
Leaves aqueous extract 250 94.2 ± 0.3 a 68.9 ± 0.7 d 82.4 ± 1.9 c
Flower aqueous extract 100 33.1 ± 1.1 g 71.4 ± 0.7 d 57.3 ± 2.2 e
Flower aqueous extract 250 66.1 ± 1.4 d 91.2 ± 0.0 b 90.0 ± 0.7 b
LSD (0.05) 3.3 5.1 3.8
Values are means of three replications ± SE. Means with the same letter are not significantly different at p60.05.
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