Chemical composition and larvicidal activity of edible plant-derived essential oils against the pyrethroid-susceptible and -resistant strains of Aedes aegypti (Diptera: Culicidae).
ABSTRACT The chemical compositions and larvicidal potential against mosquito vectors of selected essential oils obtained from five edible plants were investigated in this study. Using a GC/MS, 24, 17, 20, 21, and 12 compounds were determined from essential oils of Citrus hystrix, Citrus reticulata, Zingiber zerumbet, Kaempferia galanga, and Syzygium aromaticum, respectively. The principal constituents found in peel oil of C. hystrix were beta-pinene (22.54%) and d-limonene (22.03%), followed by terpinene-4-ol (17.37%). Compounds in C. reticulata peel oil consisted mostly of d-limonene (62.39%) and gamma-terpinene (14.06%). The oils obtained from Z. zerumbet rhizome had alpha-humulene (31.93%) and zerumbone (31.67%) as major components. The most abundant compounds in K. galanga rhizome oil were 2-propeonic acid (35.54%), pentadecane (26.08%), and ethyl-p-methoxycinnamate (25.96%). The main component of S. aromaticum bud oil was eugenol (77.37%), with minor amounts of trans-caryophyllene (13.66%). Assessment of larvicidal efficacy demonstrated that all essential oils were toxic against both pyrethroid-susceptible and resistant Ae. aegypti laboratory strains at LC50, LC95, and LC99 levels. In conclusion, we have documented the promising larvicidal potential of essential oils from edible herbs, which could be considered as a potentially alternative source for developing novel larvicides to be used in controlling vectors of mosquito-borne disease.
- [show abstract] [hide abstract]
ABSTRACT: Dichloromethane and methanol extracts of 13 Zingiberaceae species from the Alpinia, Costus and Zingiber genera were screened for antimicrobial and antioxidant activities. The antimicrobial activity of most of the extracts was antibacterial with only the methanol extract of Costus discolor showing very potent antifungal activity against only Aspergillus ochraceous (MID, 15.6 microg per disc). All the extracts showed strong antioxidant activity comparable with or higher that of alpha-tocopherol.Journal of Ethnopharmacology 11/2000; 72(3):403-10.. · 2.76 Impact Factor
- [show abstract] [hide abstract]
ABSTRACT: Mosqutio larvicidal activity of Chamaecyparis obtusa leaf-derived materials against the 4th-stage larvae of Aedes aegypti (L.), Ochlerotatus togoi (Theobald), and Culex pipiens pallens (Coquillett) was examined in the laboratory. A crude methanol extract of C. obtusa leaves was found to be active (percent mortality rough) against the 3 species larvae; the hexane fraction of the methanol extract showed a strong larvicidal activity (100% mortality) at 100 ppm. The bioactive component in the C. obtusa leaf extract was characterized as beta-thujaplicin by spectroscopic analyses. The LC50 value of beta-thujaplicin was 2.91, 2.60, and 1.33 ppm against Ae. aegypti, Oc. togoi, and Cx. pipiens pallens larvae. This naturally occurring C. obtusa leaves-derived compound merits further study as a potential mosquito larval control agent or lead compound.Journal of the American Mosquito Control Association 01/2006; 21(4):400-3. · 0.76 Impact Factor
- [show abstract] [hide abstract]
ABSTRACT: Methanolic extracts of leaves and seeds from the chinaberry tree, Melia azedarach L. (Meliaceae) was tested against mature and immature mosquito vector Anopheles stephensi Liston (Diptera) under laboratory condition. The extract showed strong larvicidal, pupicidal, adulticidal, antiovipositional activity, repellency and biting deterency. The M. azedarach seed and leaf extracts were used to determine their effect on A. stephensi adults and their corresponding oviposition and consequent adult emergence in comparison with the control. The seed extracts showed high bioactivity at all doses, while the leaf extracts proved to be active, only in the higher dose. Results obtained from the laboratory experiment showed that the seed extracts suppressed the pupal and adult activity of A. stephensi even at low dose. In general, first and second instar larvae were more susceptible to both leaves and seed extracts. Clear dose-response relationships were established with the highest dose of 2% plant extract evoking 96% mortality. Entire development of A. stephensi was inhibited by M. azedarach treatment. Less expensive (less than 0.50 US dollars per 1 kg seed), naturally accruing bio-pesticide could be an alternative for chemical pesticides.Bioresource Technology 08/2006; 97(11):1316-23. · 4.75 Impact Factor
106 Journal of Vector Ecology June 2010
Chemical composition and larvicidal activity of edible plant-derived essential oils
against the pyrethroid-susceptible and -resistant strains of Aedes aegypti
Nataya Sutthanont, Wej Choochote, Benjawan Tuetun, Anuluck Junkum, Atchariya Jitpakdi,
Udom Chaithong, Doungrat Riyong, and Benjawan Pitasawat
Department of Parasitology, Faculty of Medicine, Chiang Mai University, Chiang Mai 50200, Thailand
Received 15 November 2009; Accepted 16 December 2009
ABSTRACT: The chemical compositions and larvicidal potential against mosquito vectors of selected essential oils obtained
from five edible plants were investigated in this study. Using a GC/MS, 24, 17, 20, 21, and 12 compounds were determined
from essential oils of Citrus hystrix, Citrus reticulata, Zingiber zerumbet, Kaempferia galanga, and Syzygium aromaticum,
respectively. The principal constituents found in peel oil of C. hystrix were β-pinene (22.54%) and d-limonene (22.03%),
followed by terpinene-4-ol (17.37%). Compounds in C. reticulata peel oil consisted mostly of d-limonene (62.39%) and
γ-terpinene (14.06%). The oils obtained from Z. zerumbet rhizome had α-humulene (31.93%) and zerumbone (31.67%) as
major components. The most abundant compounds in K. galanga rhizome oil were 2-propeonic acid (35.54%), pentadecane
(26.08%), and ethyl-p-methoxycinnamate (25.96%). The main component of S. aromaticum bud oil was eugenol (77.37%),
with minor amounts of trans-caryophyllene (13.66%). Assessment of larvicidal efficacy demonstrated that all essential oils
were toxic against both pyrethroid-susceptible and resistant Ae. aegypti laboratory strains at LC50, LC95, and LC99 levels.
In conclusion, we have documented the promising larvicidal potential of essential oils from edible herbs, which could be
considered as a potentially alternative source for developing novel larvicides to be used in controlling vectors of mosquito-
borne disease. Journal of Vector Ecology 35 (1): 106-115. 2010.
Keyword Index: Mosquito larvicide, Citrus hystrix, Citrus reticulata, Zingiber zerumbet, Kaempferia galanga, Syzygium
Diseases transmitted by blood-feeding mosquitoes,
such as dengue fever, dengue hemorrhagic fever, Japanese
encephalitis, malaria, and filariasis, are increasing in
prevalence, particularly in tropical and subtropical zones.
To control mosquitoes and mosquito-borne diseases,
which have a worldwide health and economic impacts,
synthetic insecticide-based interventions are still necessary,
particularly in situations of epidemic outbreak and sudden
increases of adult mosquitoes (Yaicharoen et al. 2005,
Nathan et al. 2006). However, the indiscriminate use of
conventional insecticides is fostering multifarious problems
like widespread development of insecticide resistance, toxic
hazards to mammals, undesirable effects on non-target
organisms, and environmental pollution (Yang et al. 2002,
Kiran et al. 2006, Senthikumar et al. 2008). Each year, larger
quantities of synthetic insecticides are applied, leading to
increased dangers for humans and other organisms and
progressively greater environmental damage.
Even though plants and their preparations were the
only pest management agents available before the advent of
synthetic organic chemicals, only a few insecticides of plant
origin are now commercially available. Furthermore, most of
them have lower and shorter-lived efficacy than the synthetic
substances. Due to the high degree of biodegradation,
however, plant-derived bioproducts are currently attractive
as replacements for synthetic insecticides or for use in
integrated management programs to minimize human
health hazards and reduce the accumulation of harmful
residues in the environment. Furthermore, insect resistance
to mosquitocidal botanical agents has not previously been
documented (Shaalan et al. 2005).
Numerous products of botanical origin, especially
essential oils, have received considerable renewed attention
as potent bioactive compounds against various species of
mosquitoes. Due to the fact that application of adulticides
may only temporarily diminish the adult population (El Hag
et al. 1999, 2001), a more efficient and attractive approach
in mosquito control programs is to target the larval stage
in their breeding sites with larvicides (Amer and Mehlhorn
2006a, Knio et al. 2008). Essential oils are potentially suitable
for application in larval control management because
they constitute a rich source of bioactive compounds that
are effective and naturally biodegradable into non-toxic
products (Lucia et al. 2007, Cheng et al. 2008, 2009a).
Considerable research on the larvicidal potential of
volatile oils against Aedes aegypti mosquitoes has been
conducted and the findings contribute an insight into the
possibility of developing novel larvicides from essential
oils for use in mosquito control programs. The promising
essential oils, with larvicidal activity demonstrating LC50
Vol. 35, no. 1 Journal of Vector Ecology 107
ranging between 1-258.5 ppm, are derived from a large
number of plants, including Cymbopogon proximus, Lippia
multiflora, and Ocimum canum (Bassolé et al. 2003);
Ipomoea cairica (Thomas et al. 2004); Juniperus macropoda
and Pimpinella anisum (Prajapati et al. 2005); Citrus
bergamia, Cuminum myrrham, and Pimenta racemosa (Lee
2006); Cinnamomum camphora, Boswellia carteri, Anethum
graveolens, and Myrtus communis (Amer and Mehlhorn
2006a,b); Chloroxylon swietenia (Kiran et al. 2006); Carum
carvi, Apium graveolens, Foeniculum vulgare, Zanthoxylum
limonella, and Curcuma zedoaria (Pitasawat et al. 2007);
Zanthoxylum armatum (Tiwary et al. 2007); Eucalyptus
camaldulensis and E. urophylla (Cheng et al. 2009b).
Thailand is a source of a large diversity of medicinal
herbs. It is reassuring to ascertain plant-derived products
that are fully potential, safe, and eco-friendly. The edible
plants used as vegetables, spices, and traditional medicine
are therefore encouraging targets. Citrus plants such as Citrus
hystrix DC and C. reticulata Blanco are familiar through use
of their fruit for culinary purposes and traditional medicine
(Gimlette and Thomson 1983, Yeung 1985). Apart from use
as food flavoring and appetizer, Zingiber zerumbet Smith is
also commonly used in folkloric medicine (Burkill 1966,
Habsah et al. 2000). In Thailand and China, the rhizome
of Kaempferia galanga Linn is well-known for its popular
use as a food spice and traditional medicine (Huang et al.
2008). The flower bud of Syzygium aromaticum Linn is a
well-known food flavor in exotic food preparations and a
popular remedy in the traditional medicines of Australia
and Asian countries (Gurib-Fakim 2006).
The present study attempted to investigate the chemical
composition and larvicidal efficacy of essential oils derived
from five edible plants against both the pyrethroid-susceptible
and -resistant strains of the mosquito Ae. aegypti with the
purpose of identifying effective indigenous bioproducts to
control the vector of mosquito-borne diseases, particularly
in cases where the vector’s susceptibility to conventional
synthetics is decreasing.
MATERIALS AND METHODS
Plant materials and essential oil isolation
Five plant species, including Citrus hystrix DC., Citrus
reticulata Blanco., Zingiber zerumbet Smith., Kaempferia
galanga Linn., and Syzygium aromaticum Linn. (Table
1) were commercially obtained from traditional herb
suppliers in Chiang Mai province. Taxonomic identification
of the plants was performed by botanists and taxonomists
in the Department of Biology, Faculty of Science, Chiang
Mai University, Thailand. A voucher specimen of each
plant was deposited at the Department of Parasitology,
Faculty of Medicine, Chiang Mai University, Thailand.
Each plant material was shade-dried at room temperature,
mechanically ground by an electrical blender, and steam
distilled at 100° C for at least three h to obtain essential
oils. The oil layer was separated from the aqueous phase,
dried over anhydrous sodium sulfate (Na2So4), and kept in
an amber-colored bottle under refrigeration at 4° C until
further chemical analysis and larvicidal bioassays. In each
case, the yield of oil was averaged over three experiments
and calculated according to the dry weight of the plant
Gas chromatography-mass spectrometry (GC/MS)
Analysis of essential oils was performed by gas
chromatography coupled with mass spectrometry (GC/
MS) in a Hewlett-Packard 6850 gas chromatograph (Agilent
Technologies) equipped with a split-splitless injector and
HP-5MS (30 m x 0.25 mm ID and 0.25 µm film thickness)
columns directly coupled to a quadrupole mass selective
detector, MSD 5973 (Agilent Technologies). The injector
temperature was set at 250° C and the oven temperature
was initially at 50° C, and programmed to reach 230° C at
the rate of 6° C/min and held at 230° C for 10 min, then
increased from 230° C to 250° C at the rate of 10° C/min.
Helium was used as the carrier gas with a flow rate of 1.0
ml/min. The sample (0.2 µl) was injected neat with a split
ratio of 250:1. The mass spectrometer (MSD 5973) was
operated in the electron impact (EI) mode at 70eV. The ion
source and quadrupole temperatures were set at 230° C and
150° C, respectively. The oil components were identified by
comparison with standards, by spiking and on the basis of
their mass spectral fragmentation using the NIST 05, NIST
98, Wiley 7N, and Wiley 275 GC/MS libraries. Percentage
of the identified compound was computed from a total ion
Mosquito populations tested in the larvicidal bioassays
comprised both pyrethroid-susceptible and -resistant Ae.
aegypti laboratory strains. The pyrethroid susceptible Ae.
aegypti colony was established from specimens collected
in Muang district, Chiang Mai province, and maintained
continuously since 1995. The pyrethroid-resistant Ae.
aegypti colony was collected from various places in Mae
Tang district, Chiang Mai province, and maintained under
selective pressure (Chareonviriyaphap et al. 2002) to establish
a pyrethroid-resistant laboratory colony. Both pyrethroid-
susceptible and -resistant Ae. aegypti were maintained and
reared separately without exposure to any insecticides or
pathogens in an insectary (6.2 x 7.3 x 3 m) at 25±2ο C and
80%±10% RH under a 14:10 light-and-dark cycle, following
standard operating procedures for mosquito maintenance
(Limsuwan et al. 1987). Tests of susceptibility of adult Ae.
aegypti mosquitoes to the synthetic pyrethroids, permethrin
and lambdacyhalothrin, were regularly conducted using
WHO test kits with some modifications (WHO 1998).
Newly molted 4th instar Ae. aegypti larvae were continuously
available for the larvicidal bioassays.
Mosquito larvicidal bioassays
The mosquito larvicidal assays were carried out
under laboratory conditions by a slight adaptation of the
standard protocol recommended by the World Health
Organization (WHO 1981). Each essential oil was dissolved
108 Journal of Vector Ecology June 2010
in dimethylsulphoxide (DMSO) to prepare graded
concentrations of tested material. In preparing a series of
aqueous solutions with different concentrations of tested
oil, 1 ml of DMSO solution containing the desired essential
oil was completely mixed with 249 ml of distilled water in
an enamel bowl of 10 cm in diameter and 8 cm in depth.
Four batches of 25 early 4th instar larvae of Ae. aegypti were
maintained in 250 ml of aqueous solutions with the final
total number of 100 larvae for each concentration. A total of
five essential oils were tested in this manner with a range of
concentrations (four to six concentrations, ppm), yielding a
range of 0-100% mortality. Control tests receiving DMSO-
distilled water were performed in parallel for comparison.
Mortality and survival of larvae were determined after
24 h of exposure and the larvae were starved within this
period. Observations were also made on the behavior of
larvae. Larvae were considered dead when they did not
respond to stimuli such as probing with a needle in the
siphon or cervical region. Moribund larvae were those
incapable of rising to the surface of the water (within a
reasonable period of time) or showing a characteristic
diving reaction when the water was disturbed. They might
also show discoloration, unnatural positions, tremors,
incoordination, or rigor. The moribund and dead larvae
in each concentration were combined in quadruplicate
and expressed as percentage mortalities. Every bioassay
was carried out in environmentally controlled conditions
(temperature ∼ 25±2ο C; humidity ∼ 80%±10% RH; 14-h
light and 10-h dark cycle), and replicated four times with
mosquitoes from different rearing batches. The percentage
mortality was reported from the average of four replicates.
It was important to obtain no less than three mortality
counts of between 10% and 90%. In cases where the control
mortality was between 5-20%, the observed percentage
mortality (%M) was corrected by Abbott’s formula (Abbott
%M = % test mortality - % control mortality x 100
100 - % control mortality
Data for larvicidal potential were analyzed by means
of computerized probit analysis (Harvard Programming;
Hg1, 2), yielding the lethal concentrations LC
and 95% confidence intervals (CI) of upper and lower
confidence levels. Significant differences were determined
by comparing the CI of each plant oil.
RESULTS AND DISCUSSION
The yields of volatile oils ranged from a minimum of
0.30% to a maximum of 3.36% (v/w) according to dry weight
(Table 1). The highest oil content was found in C. hystrix
(3.36%), followed by S. aromaticum (1.50%), C. reticulata
(1.40%), K. galanga (0.76%), and Z. zerumbet (0.30%). It is
generally known that the yield of essential oil depends not
only on the plant species and their climatic or geographical
areas, but also other variables such as method of extraction
and plant-related factors, including parts of plant, rearing
condition, maturation of the harvested plant, and plant
storage or preservation (Vieira and Simon 2000, Tawatsin
et al. 2001). In order to achieve the best yield, it is therefore
necessary to establish the most appropriate combination of
these variable factors. However, in addition to the yield of
Table 1. Ethnobotanical data, physical characteristics, and percentage yields (% Yield) of essential oils derived from five
Family & Botanical name
Citrus hystrix DC
Citrus reticulata Blanco
Vol. 35, no. 1 Journal of Vector Ecology 109
Table 2. Percentage composition of essential oils from five edible plants.
C. reticulataZ. zerumbet K. galanga S. aromaticum
5.22 31.93 1.53
110 Journal of Vector Ecology June 2010
Table 3. Larvicidal activity of edible plant-derived essential oils against 4th instar larvae of pyrethroid-susceptible
Concentration of plant oil
(95% CI, ppm)
Vol. 35, no. 1 Journal of Vector Ecology 111
Figure 1. GC-MS total ion chromatograms for essential oils of five plants.
112 Journal of Vector Ecology June 2010
essential oil, much consideration was given to the quality
and quantity of chemical constituents, particularly the
major active ingredients.
In this study, GC/MS characterization was performed
to show the profile of constituents in the selected essential
oils, of which gas chromatograms and percentage
compositions are presented in Figure 1 and Table 2,
respectively. A total of 68 compounds were identified
from five essential oils, representing 96.01- 100% of the oil
obtained. The essential oil of C. hystrix peel contained 24
identified components, amounting to 99.52% of the whole
oil with β-pinene (22.54%) and d-limonene (22.03%) as the
principal constituents, followed by terpinene-4-ol (17.37%),
together with trace amounts of α-terpineol (6.29%) and
sabinene (5.49%). For C. reticulata peel oil, 17 compounds
were identified, representing 100.00% of the whole oil with
the rich constituents of d-limonene (62.39%), followed by
γ-terpinene (14.06%), with minor contents of 1-methyl-2-
(1-methylethyl) (6.46%), α-humulene (5.22%), and methyl-
n-methyl anthranilate (3.25%). The rhizome oil of Z.
zerumbet showed the presence of 20 compounds, accounting
for 96.01% of the whole oil with α-humulene (31.93%) and
zerumbone (31.67%) as the main constituents, followed by
minor quantities of o-menth-8-ene (8.46%), santolina triene
(5.38%), β-caryophyllene (3.36%), and camphor (3.05%). A
total of 21 compounds were identified in K. galanga rhizome
oil, corresponding to 98.89% of the total oil. The most
abundant compounds were 2-propeonic acid (35.54%),
pentadecane (26.08%), and ethyl-p-methoxycinnamate
(25.96%), whereas 3-carene (2.47%) and eucalyptol (2.12%)
were minor constituents. Twelve compounds constituting
100.00% of all the volatile compositions were characterized
from S. aromaticum bud oil containing the chief constituent
of eugenol (77.37%), followed by minor amounts of trans-
caryophyllene (13.66%) and eugenol acetate (4.60%).
In the larvicidal assessment, all essential oils
demonstrated efficacy in both the pyrethroid-susceptible
and -resistant strains of Ae. aegypti with dose dependent
and different degrees among plant species (Tables 3 and
4). When exposed to the higher oil concentrations, more
larvae showed toxic symptoms that led to an increase in
mortality values. Correspondingly, the treated larvae tended
to show toxic symptoms and die earlier at increasing oil
concentrations. These findings suggest that concentrations
of test substance affected degree of toxicity, mortality
speed, and mortality rates. Although toxic symptoms in
larvae treated with each essential oil were observed during
different periods of time, the symptoms in larvae treated
by these oils seemed to be similar, depending on dosage
and oil variety. The incapacitated larvae showed abnormal
behaviors such as restlessness, sluggishness, and coiling
movement, and subsequently settled at the bottom of the
bowl with abnormal wagging, tremors, convulsions, and
paralysis, and later died slowly. However, no mortality was
observed in the control groups.
Even though effects on both strains of Ae. aegypti were
relatively similar, all the oils tested proved to be slightly
more toxic against the pyrethroid-susceptible strain than
the resistant one. The highest potential was established
from C. reticulata, followed by C. hystrix, Z. zerumbet, K.
galanga, and S. aromaticum, with an LC50 of 15.42, 30.07,
48.88, 53.64, and 124.69 ppm, respectively, in the pyrethroid
susceptible strain, and 19.38, 34.78, 53.08, 59.03, and 143.89
ppm, respectively, in the pyrethroid-resistant strain. The
susceptibility to essential oils between the two strains of Ae.
aegypti was slightly different but statistically significant. This
performance has been initially observed and documented
in our study. However, the relevance of these findings,
which are probably due to mosquito tolerance or resistance
to botanical agents, cannot be explained at the moment and
requires more extensive studies.
In Thailand, due to the higher cost and lower efficacy
compared with conventional synthetics, the plant-derived
mosquitocides have been subjected to use in the vector
control programs with lower frequency, for shorter periods,
and on smaller scales. However, the increasing utilization
of plants and plant products for pest management in
agriculture possibly led to the cross-resistance effect. The
similarity in chemical structure and/or mechanism of
action between the pyrethroid insecticides and used plant
products might be a key to the development of tolerance
or resistance in natural populations of mosquitoes. In order
to clarify this suspicion, isolation and identification of the
active ingredients responsible for such larvicidal activity
need to be performed.
In recent years, the active insecticidal compounds
isolated from plants have received much attention due
to their pronounced larvicidal efficacy. The bioactive
component, β-thujaplicin, derived from Chamaecyparis
obtusa leaf extract demonstrated strong larvicidal potential
against Ae. aegypti, Ae. togoi, and Culex pipiens pallens,
with LC50 of 2.91, 2.60, and 1.33 ppm, respectively (Jang
et al. 2005). Larvicidal investigation of Eucalyptus grandis
essential oil and its major components on Ae. aegypti
revealed that the most effective was β-pinene, followed
by α-pinene, and 1,8-cineole with the LC50 of 12.1, 15.4
ppm, and 57.2 ppm, respectively (Lucia et al. 2007).
The essential oils of Cryptomeria japonica leaf and their
effective constituents, including α–terpinene, γ-terpinene,
ρ-cymene, 3-carene, terpinolene, and β–myrcene, provided
an excellent larvicidal effect against both Ae. aegypti and Ae.
albopictus, with an LC50 below 40 µg/ml. Among the pure
constituents, 3-carene and terpinolene exhibited the best
inhibitory action against Ae. aegypti (LC50 = 25.3 µg/ml)
and Ae. albopictus (LC50 = 22.0 µg/ml), respectively (Cheng
et al. 2008, 2009a).
The toxicities of ethyl cinnamate and ethyl
p-methoxycinnamate identified in K. galanga rhizome
and another 12 known compounds were evaluated against
3rd instar larvae of laboratory-reared Cx. pipiens pallens,
Ae. aegypti, and Ae. togoi, and field-collected Cx. pipiens
pallens. Results were compared with those for fenthion and
temephos (Kim et al. 2008). Ethyl p-methoxycinnamate, the
most effective of the plant-derived compounds (LC50=12.3-
20.7 mg/l), was found to be less toxic than either fenthion
(LC50=0.0096-0.021 mg/l) or temephos (LC50=0.0039-
Vol. 35, no. 1 Journal of Vector Ecology 113
Figure 2. Larvicidal activity of five essential oils against the 4th instar larvae of pyrethroid susceptible and resistant strains of
114 Journal of Vector Ecology June 2010
0.0079 mg/l). Ethyl cinnamate and 3-carene were highly
active against Cx. pipiens pallens (LC50=24.1 and 21.6 mg/l,
respectively) but less toxic to Ae. aegypti and Ae. togoi.
Variations in toxicity of essential oils against different
species of mosquitoes are common (Sukumar et al. 1991,
Amer and Mehlhorn 2006a), due to qualitative and
quantitative variations of chemical constituents. Interestingly,
the active larvicidal compounds in these works, including
α-pinene, β-pinene, 1,8-cineole, α–terpinene, γ-terpinene,
terpinolene, 3-carene, β–myrcene, ethyl cinnamate, and
ethyl p-methoxycinnamate, were also detected in the
essential oils investigated in this study. Therefore, other
compounds such as d-limonene, α-humulene, zerumbone,
2-propeonic acid, pentadecane, and eugenol identified as
the major components in the effective essential oils derived
from C. hystrix, C. reticulata, Z. zerumbet, K. galanga, or S.
aromaticum should not be neglected.
Isolation and purification of the active compounds that
might be responsible for the larvicidal activity against Ae.
aegypti could be an important next step in the development
of novel mosquitocidal agents. Production of larvicides from
the locally available edible plants, which could be a new
acceptable alternative to employ in the water supplies for
drinking and daily use, may lead to decreasing dependence
on imported synthetic insecticides and be beneficial for
developing countries such as Thailand. Despite essential
oils likely having less potential than synthetic pyrethroids,
their natural biodegradation and remarkable activity on
pyrethroid-resistant mosquitoes make them promising
candidates for further study in controlling dengue and other
This work is supported by the Faculty of Medicine
Research Fund, Faculty of Medicine, Chiang Mai University,
Thailand. The authors acknowledge Assoc. Prof. Dr. Chusie
Trisonthi and Assist. Prof. Paritat Trisonthi, botanists
and taxonomists at the Department of Biology, Faculty of
Science, Chiang Mai University, Thailand, for their kindness
in identification of the plant samples.
Abbott, W.S. 1925. A method of computing the effectiveness
of an insecticide. J. Econ. Entomol. 18: 265-266.
Amer, A. and H. Mehlhorn. 2006a. Larvicidal effects of
various essential oils against Aedes, Anopheles, and
Culex larvae (Diptera: Culicidae). Parasitol. Res. 99:
Amer, A. and H. Mehlhorn. 2006b. Persistency of larvicidal
effects of plant oil extracts under different storage
conditions. Parasitol. Res. 99: 478-490.
Bassolé, I.H., W.M. Guelbeogo, R. Nébié, C. Costantini, N.
Sagnon, Z.I. Kabore, and S.A. Traoré. 2003. Ovicidal
and larvicidal activity against Aedes aegypti and
Anopheles gambiae complex mosquitoes of essential
oils extracted from three spontaneous plants of Bukina
Faso. Parasitologia 45: 23-26.
Burkill, I.H. 1966. A Dictionary of the Economic Products
of the Malay Peninsula. Ministry of Agriculture and
Cooperative, Kuala Lumpur.
Chareonviriyaphap, T., P. Rongnoparut, and P. Juntarumporn.
2002. Selection for pyrethroid resistance in a colony
of Anopheles minimus species A, a malaria vector in
Thailand. J. Vector Ecol. 27: 222-229.
Cheng, S.S., M.T. Chua, E.H. Chang, C.G. Huang, W.J.
Chen, and S.T. Chang. 2009a. Variations in insecticidal
activity and chemical composition of leaf essential oils
from Cryptomeria japonica at different ages. Bioresorce
Technol. 100: 465-470.
Cheng, S.S., C.G. Huang, W.J. Chen, Y.H. Kuo, and S.T.
Chang. 2008. Larvicidal activity of tectoquinone
isolation from red heartwood-type Cryptomeria
japonica against two mosquito species. Bioresource
Technol. 99: 3617-3622.
Cheng, S.S., C.G. Huang, Y.J. Chen, J.J. Yu, W.J. Chen,
and S.T. Chang. 2009b. Chemical compositions and
larvicidal activities of leaf essential oils from two
eucalyptus species. Bioresource Technol. 100: 452-456.
El Hag, E.A., A.H. Nadi, and A.A. Zaitoon. 1999. Toxic and
growth retarding effects of three plant extracts on Culex
pipiens larvae (Diptera: Culicidae). Phytother. Res. 13:
El Hag, E.A., A-E. Rahman, H. El-Nadi, and A.A. Zaitoon.
2001. Effects of methanolic extracts of neem seeds
on egg hatchability and larval development of Culex
pipiens mosquitoes. Indian Vet. J. 78: 199-201.
Gimlette, J.D. and H.W. Thomson. 1983. A Dictionary of
Malayan Medicine. Oxford University Press, Kuala
Gurib-Fakim, A. 2006. Medicinal plants: traditions of
yesterday and drugs of tomorrow. Mol. Aspects Med.
Habsah, M., M. Amran, M.M. Mackeen, N.H. Lajis, H.
Kikuzaki, N. Nakatani, A.A. Rahman, A. Ghafar,
and A.M. Ali. 2000. Screening of Zingiberaceae
extracts for antimicrobial and antioxidant activities. J.
Ethnopharmacol. 72: 403-410.
Huang, L., T. Yagura, and S. Chen. 2008. Sedative activity of
hexane extract of Keampferia galanga L. and its active
compounds. J. Ethnopharmacol. 30: 123-125.
Jang, Y.S., J.H. Jeon, and H.S. Lee. 2005. Mosquito
larvicidal activity of active constituent derived from
Chamaecyparis obtusa leaves against 3 mosquito
species. J. Am. Mosq. Contr. Assoc. 21: 400-403.
Kim, N.J., S.G. Byun, J.E. Cho, K. Chung, and Y.J. Ahn.
2008. Larvicidal activity of Kaempferia galanga rhizome
phenylpropanoids towards three mosquito species.
Pest. Manag. Sci. 64: 857-862.
Kiran, R.S., K. Bhavani, S.P. Devi, R.B.R. Rao, and J.K. Reddy.
2006. Composition and larvicidal activity of leaves and
stem essential oils of Chloroxylon swietenia DC. against
Aedes aegypti and Anopheles stephensi. Bioresource
Technol. 97: 2481-2484.
Knio, K.M., J. Usta, S. Dagher, H. Zournajian, and S.
Vol. 35, no. 1 Journal of Vector Ecology 115
Kreydiyyeh. 2008. Larvicidal activity of essential oils
extracted from commonly used herbs in Lebanon
against the seaside mosquito, Ochlerotatus caspius.
Bioresource Technol. 99: 763-768.
Lee, H.S. 2006. Mosquito larvicidal activity of aromatic
medicinal plant oils against Aedes aegypti and Culex
pipiens pallens. J. Am. Mosq. Contr. Assoc. 22: 292-
Limsuwan, S., Y. Rongsriyam, V. Kerdpibule, C.
Apiwathnasorn, G.L. Chiang, and W.H. Cheong. 1987.
Rearing techniques for mosquitoes. In: S. Sucharit and
S. Supavej (eds.) Practical Entomology. Malaria and
Filariasis. Museum and Reference Centre, Mahidol
Lucia, A., G.A. Audino, E. Seccacini, S. Licastro, E. Zerba,
and H. Masuh. 2007. Larvicidal effect of Eucalyptus
grandis essential oil and turpentine and their major
components on Aedes aegypti larvae. J. Am. Mosq.
Contr. Assoc. 3: 299-303.
Nathan, S.S., K. Kalaivani, and K. Sehoon. 2006. Effects of
Dysoxylum malabaricum Bedd. (Meliaceae) extract on
the malarial vector Anopheles stephensi Liston (Diptera:
Culicidae). Bioresource Technol. 97: 2077-2083.
Pitasawat, B., D. Champakaew, W. Choochote, A. Jitpakdi,
U. Chaithong, D. Kanjanapothi, E. Rattanachanpichai,
P. Tippawangkosol, D. Riyong, B. Tuetun, and D.
Chaiyasit. 2007. Aromatic plant-derived essential oil: an
alternative larvicide for mosquito control. Fitoterapia
Prajapati, V., A.K. Tripathi, K.K. Aggarwal, and S. Khanuja.
2005. Insecticidal, repellent and oviposition-deterrent
activity of selected essential oils against Anopheles
stephensi, Aedes aegypti and Culex quinquefasciatus.
Bioresource Technol. 96: 1749-1757.
Senthilkumar, A., K. Kannathasan, and V. Venkatesalu.
2008. Chemical constituents and larvicidal property of
the essential oil of Blumea mollis (D. Don) Merr. against
Culex quinquefasciatus. Parasitol. Res. 103: 959-962.
Shaalan, E., D. Canyon, M.W. Faried, H. Abdel-Wahab, and
A. Mansour. 2005. A review of botanical phytochemicals
with mosquitocidal potential. Environ. Int. 31: 1149-
Sukumar, K., M.J. Perich, and L.R. Boobar. 1991. Botanical
derivatives in mosquito control: a review. J. Am. Mosq.
Contr. Assoc. 7: 210-237.
Tawatsin, A., S.D. Wratten, R.R. Scott, U. Thavara, and Y.
Techadamrongsin. 2001. Repellency of volatile oils
from plants against three mosquito vectors. J. Vector
Ecol. 26: 76-82.
Thomas, T.G., S. Rao, and S. Lai. 2004. Mosquito larvicidal
properties of essential oil of an indigenous plant,
Ipomoea cairica Linn. Jpn. J. Infect. Dis. 43: 176-177.
Tiwary, M., S.N. Naik, D.K. Tewary, P.K. Mittal, and S.
Yadav. 2007. Chemical composition and larvicidal
activities of the essential oïl of Zanthoxylum armatum
DC. (Rutaceae) against three mosquito vectors. J.
Vector Borne Dis. 3: 198-204.
Vieira, R.F. and J.E. Simon. 2000. Chemical characterization
of basil (Ocimum spp.) found in the markets and used
in traditional medicine in Brazil. Econ. Bot. 54: 207-
WHO. 1981. Instructions for determining the susceptibility
or resistance of mosquito larvae to insecticides. World
Health Organization, Geneva, P 6 WHO/VBC/81.807.
WHO. 1998. Test procedures for insecticide resistance
monitoring in malaria vectors, bio-efficacy and
persistence of insecticides on treated surfaces. World
Health Organization, Geneva WHO/CDS/CPC/
Yaicharoen, R., R. Kiatfuengfoo, T. Chareonviriyaphap, and
P. Rongnoparut. 2005. Characterization of deltamethrin
resistance in field populations of Aedes aegypti in
Thailand. J. Vector Ecol. 30: 144-150.
Yang, Y.C., S.G. Lee, H.K. Lee, M.K. Kim, S.H. Lee, and H.S.
Lee. 2002. A piperidine amide extracted from Piper
longum L. fruit shows activity against Aedes aegypti
mosquito larvae. J. Agric Food Chem. 50: 3765-3767.
Yeung, H.C. 1985. Handbook of Chinese Herbs and Formulas.
Institute of Chinese Medicine, Los Angeles.