Parasitol Res
DOI 10.1007/s00436-013-3528-7
ORIGINAL PAPER
Impact of Argemone mexicana extracts on the cidal,
morphological, and behavioral response of dengue vector,
Aedes aegypti L. (Diptera: Culicidae)
Radhika Warikoo & Sarita Kumar
Received: 23 June 2013 / Accepted: 26 June 2013
# Springer-Verlag Berlin Heidelberg 2013
Abstract The larvicidal, behavioral, and morphological re-
sponse of dengue vector, Aedes aegypti treated with delete-
rious weed, Argemone mexicana, was explored. The
1,000 ppm extracts of A. mexicana leaf, stem, and roots
prepared in five different solvents (petroleum ether, hexane,
benzene, acetone, and ethanol) were screened for their larvi-
cidal activity against dengue vector establishing the efficacy
of petroleum ether and hexane extracts. Other extracts, un-
able to give 100 % mortality, were considered ineffective and
discarded from further study. Larvicidal bioassay conducted
with selected extracts confirmed the higher efficacy of hex-
ane extracts exhibiting 1.1- to 1.8-fold more potential than
the petroleum ether extracts. The results further revealed 1.6-
to 2.4-fold higher efficacy of root extracts than those pre-
pared from the leaves and stem of A. mexicana. The hexane
root extract of A. mexicana was found to be the most effec-
tive larvicide with LC50 value of 91.331 ppm after 24 h of
exposure causing 1.8 and 2.4 fold more toxicity as compared
to the hexane leaf and stem extracts, respectively. Prolonged
exposure of the larvae to the extracts resulted in increased
toxicity potential of the extracts. Observations of the treated
larvae revealed excitation, violent vertical, and horizontal
movements with aggressive anal biting behavior suggesting
effect of extracts on their neuromuscular system. Morpholog-
ical studies of the treated larvae revealed the demelanization
of cuticle and shrinkage of internal cuticle of anal papillae
indicating the anal papillae as the probable action sites of the
A. mexicana extracts. The potential of A. mexicana as new
larvicides against dengue vector are being explored.
R. Warikoo (*) : S. Kumar
Department of Zoology, Acharya Narendra Dev College,
University of Delhi, New Delhi, India
e-mail: radhikawarikoo@yahoo.com
Introduction
Mosquitoes are collectively the single most important family
of insects from the human perspective. Due to their high
potential to exploit even adverse environmental conditions,
mosquitoes can rapidly increase their population. Aedes
aegypti, the primary vector for dengue fever, dengue hemor-
rhagic fever, and yellow fever is widespread over large areas
of the tropics and subtropics. According to the World Health
Organization (WHO), about 40 % of the worlds population
is now at risk of dengue and the only way to prevent dengue
virus transmission is to combat the disease-carrying mosqui-
toes (WHO 2012). In India, official records of the Union
Health Ministry reveal that there has been a massive increase
of dengue infection in the country in 2012. The correspond-
ing figure till 2012, stood at 50,222 cases and 241 fatalities in
contrast to 18,860 cases and 169 deaths in 2011 (NVBDCP
2013).
Synthetic chemical insecticides are popularly used as the
first line of defense against mosquitoes owing to their quick
action. Over the last five decades, the indiscriminate and
frequent use of these synthetic insecticides has led to the
multifarious problems (viz., insecticide resistance, permanent
residual effect on the environment, and destabilization of the
ecosystem) and toxic hazards to human and nontarget organ-
isms (Jirakanjanakit et al. 2007; Sarwar et al. 2009). In recent
years, much effort has been focused on plant extracts or
phytochemicals as potential sources of mosquito control agents
and as a viable component of Integrated Pest Management to
prevent the inevitable toxicity and resistance issues caused by
chemical insecticides (Warikoo et al. 2011).
Phytochemicals have been reported as ecofriendly, envi-
ronmentally safe, biodegradable, and low-cost natural prod-
ucts which show target specificity. A large number of plant
extracts have been shown to have mosquitocidal or repellent

activity against mosquito vectors (Amer and Mehlhorn
2006a, b; Rahuman et al. 2008a, b; Kumar et al. 2012a),
though the insecticidal effects of plant chemicals vary not
only according to plant species, mosquito species, and plant
parts, but also with extraction methodology. Phytochemicals
do not pose any threat to the environment and also possess
multiple effects against vector mosquitoes which include
growth regulation, fecundity suppression, male sterility, lar-
vicidal, ovicidal, and oviposition activity mostly as deter-
rence (Kumar et al. 2010, 2011a, b). Nevertheless, the mos-
quito control potential of weed plants found in vast areas on
plains as well as on hilly regions has not been attempted at
large scale (Raj Mohan and Ramaswamy 2007; Kumar et al.
2012b). Only a few reports are available regarding the po-
tential of certain weeds as larvicidal, ovicidal, and oviposi-
tion deterrent agent against A. aegypti (Sathish and
Maneemegalai 2008; Kumar et al. 2011a, 2012b).
Argemone mexicana is a common weed which is used in
the traditional system of medicine to cure malarial fever and
ulcers. Its seeds are efficient in treating leprosy, dropsy, and
jaundice (Alagesaboopathi 2009). Certain bioactive com-
pounds derived from A. mexicana seeds have been proven
to possess chemosterilant activity including reduction in
blood meal utilization, diminished fecundity, formation of
larvalpupal or pupaladult intermediates, adult mortality, and
sterility of first-generation eggs in A. aegypti (Sakthivadivel
and Thilagavathy 2003). The available literature, however,
reveals that this plant has not been explored extensively
against mosquito vectors as larvicidal agent or by affecting
their biological characteristics. Hence, this study was under-
taken to assess the probable use of extracts prepared from
different parts of A. mexicana against dengue vector larvae
causing mortality, behavioral changes, or morphological alter-
ations. The assessment of mosquito control potential of this
weed, besides its management may be useful in promoting
research aiming at the development of new mosquito control
agent.
Materials and methods
Mosquito rearing
The present investigations employ the dengue fever mosquito,
A. aegypti, originated from fields of Delhi and surrounding
areas. The colony was maintained in an insectary at 281 C,
805 % RH and 14:10 light/day photoperiod (Warikoo et al.
2011). Wet cotton was kept on the top of each cage to provide
water for the mosquitoes. Water-soaked split raisins were kept
in the cage, mainly as a source of the food for the male
mosquitoes. Periodic blood meals were provided to female
mosquitoes for egg maturation by keeping restrained albino
rats in the cages. The eggs were collected in an enamel bowl
Parasitol Res
lined with Whatman filter paper on all the sides and half-filled
with dechlorinated tap water. The eggs were allowed to hatch
in trays filled with dechlorinated water. Larvae were fed upon
a mixture of finely ground dog biscuits and yeast in the ratio of
3:1 by weight. Care was taken to prevent formation of any
scum on the surface of water. The pupae formed were collect-
ed and transferred to the cloth cages for adult emergence.
Plant collection
Different parts of the A. mexicana plant, i.e., leaves, stems,
and roots, were collected from the surrounding areas in New
Delhi, India. The collected parts were thoroughly washed
with tap water and dried under the shade at room temperature
of 272 C separately for about 20 days. The dried parts
were then crushed, powdered, and sieved thoroughly to get
fine powder.
Preparation of the extract
The powdered plant parts, i.e., roots, stems, and leaves, were
weighed separately. Of each powdered material, 200 g was
extracted in 1,000 mL of petroleum ether, hexane, benzene,
acetone, and ethanol, separately using Soxhlet extraction
apparatus. The extraction continued for 3 days, 8 h per day,
at a temperature not exceeding the boiling point of the
solvent. The crude extracts, thus formed, were concentrated
using a vacuum evaporator at 45 C under low pressure.
After complete evaporation of the solvent, the concentrated
extracts were collected and stored in a refrigerator at 4 C as
the stock solution of 1,000 ppm for further use. This stock
solution was used to prepare the desired concentrations of
the extracts for investigating the cidal, behavioral, and mor-
phological effects against larvae of A. aegypti.
Screening of extracts for their larvicidal efficacy against A.
aegypti
The larvicidal bioassay was performed at 281 C on early
fourth instars of A. aegypti in accordance with the procedure
described by the WHO with slight modifications (WHO
2005). The graded series of all the extracts were prepared
using ethanol as the solvent. For experimental treatment,
1 ml of 1,000 ppm plant extract was added to 100 ml of
distilled water in a 250-mL glass jar. The mixture was shaken
lightly to ensure a homogeneous test solution. The early
fourth instars of A. aegypti, in batches of 20, were taken in
plastic bowls containing 99 mL of distilled water and trans-
ferred to glass jar containing distilled waterextract mixture.
Four replicates were carried out simultaneously for each
extract making a total of 80 larvae for each test. Controls
were exposed to the solvent, i.e., ethanol alone. During the
treatment period, the larvae were not provided with any food.
Parasitol Res

Table 1 Screening of different extracts (1,000 ppm) prepared from Statistical analysis of data
leaf, stem and root of A. mexicana for larvicidal activity against dengue
vector A. aegypti
The larvicidal tests with more than 20 % mortality in controls
S. no. Part used Extract Mortality after 24 h (%) and pupae formed were discarded and repeated again. If the
control mortality ranged between 5 and 20 %, it was
1 Leaf Hexane extract 100
corrected using Abbotts formula (Abbott 1925).
Petroleum ether extract 100
The data were subjected to regression analysis using com-
Benzene extract 57.5
puterized SPSS 16.0 Program. The LC50 and LC90 values with
Acetone extract 50
95 % fiducial limits were calculated in each bioassay to
Ethanol extract 70
measure difference between the test samples.
2 Stem Hexane extract 100
Petroleum ether extract 100 Behavioral studies in extract-treated A. aegypti larvae
Benzene extract 75
Acetone extract 60 During each larvicidal bioassay, the larvae were monitored
Ethanol extract 57.5 carefully for behavioral modifications, if any, caused by
3 Root Hexane extract 100 extract-mediated disruption of biological functions. The be-
Petroleum ether extract 100 havioral observations included wriggling speed, horizontal
Benzene extract 77.5 movements, vertical movements, aggregation behavior, and
Acetone extract 15 knockdown. The larval behavior was recorded and
Ethanol extract 25 photographed with Canon Power Shot SX50HS. Similar
observations were made in controls for comparison with
treated larvae.

The dead and moribund larvae were recorded after 24 h.
Similar tests were carried out with each extract against early
fourth instars of A. aegypti to assess the larvicidal efficiency
of A. mexicana.
Evaluation of larvicidal potential of selected extracts
against A. aegypti
The extracts that failed to give 80100 % larval mortality at
1,000 ppm were considered ineffective and not tested further
for larvicidal efficacy. Other extracts causing 80100 % larval
mortality at 1,000 ppm were evaluated further for larvicidal
potential. The bioassays were performed as described earlier.
Four replicates were carried out simultaneously for each assay
making a total of 80 larvae for each concentration of each
extract. Controls were exposed to the solvent, i.e., ethanol
alone. The larval mortality was recorded after 24 as well as
48 h of exposure.
Morphological studies in extract-treated A. aegypti larvae
After treatment, the dead larvae were scrutinized for mor-
phological alterations under light microscopy. Morphologi-
cal modifications in body segments including the head, tho-
rax, and abdomen, and other organs such as the eyes, anten-
nae, mouth brushes, setae, saddle, and anal gills were ob-
served, photographed, and compared with those of the con-
trols. The larvae were also observed carefully for any
changes in pigmentation pattern.
Results
The potential of leaf, stem, and root extracts of A. mexicana in
five different solvents was studied against A. aegypti larvae for
use as ecofriendly insecticides as an alternative for eco-enemy
synthetic insecticides. The results of the larvicidal tests

Table 2 Larvicidal activity of the root extracts of A. mexicana against early fourth instars of A. aegypti

Solvent Time of exposure (h) Larvicidal activity SE 2 (df) Regression coefficient

LC50 (ppm) LC90 (ppm)

Hexane 24 91.331 (79.987106.554) 156.684 (128.987224.152) 0.96 2.809 (4) 5.467

48 82.468 (71.23696.639) 153.202 (124.003226.723) 0.84 0.752 (4) 5.666
Petroleum ether 24 158.839 (132.457194.134) 339.477 (264.074507.541) 0.56 2.556 (4) 3.885
48 130.615 (110.627159.487) 256.334 (199.818401.296) 0.74 1.436 (3) 4.377

Values in parentheses indicate the lower and upper 95 % fiducial limits

 Parasitol Res

Table 3 Larvicidal activity of the leaf extracts of A. mexicana against early fourth instars of A. aegypti

Solvent Time of exposure (h) Larvicidal activity SE 2 (df) Regression coefficient

LC50 (ppm) LC90 (ppm)

Hexane 24 168.830 (139.807207.408) 377.446 (291.136569.695) 0.52 3.192 (4) 3.668

48 135.942 (112.910165.928) 300.429 (232.493458.386) 0.56 2.426 (4) 3.721
Petroleum ether 24 299.565 (244.049366.232) 696.755 (541.9881,028.454) 0.49 4.060 (4) 3.496
48 194.580 (153.497247.278) 524.32 (381.983913.834) 0.49 4.923 (3) 2.976

Values in parentheses indicate the lower and upper 95 % fiducial limits

performed against fourth instars of A. aegypti with 1,000 ppm
of different extracts prepared in five solvents, i.e., petroleum
ether, hexane, benzene, acetone, and ethanol are presented in
Table 1. The data clearly reveals the varied potential of ex-
tracts against A. aegypti with only petroleum ether and hexane
extracts as effective larvicides leading to 100 % larval mor-
tality at 1,000 ppm (Table 1). The remaining extracts proved
to be 2285 % less efficient exhibiting a range of 1578 %
larval mortality. The least effective extract was found to be the
acetone root extract whereas ethanol leaf extract and benzene
root extract exhibited moderate toxicity (Table 1). The other
extracts, except hexane and petroleum ether extracts, were not
considered for further investigations.
The larvicidal bioassays carried out with the selected ex-
tracts against A. aegypti revealed that though petroleum ether
extracts were quite effective against dengue vector larvae,
hexane extracts exhibited 1.1- to 1.8-fold more larvicidal
potential than the petroleum ether extracts, irrespective of
whether these were prepared from roots, stems, or leaves of
A. mexicana (Tables 2, 3, and 4, Fig. 1). The results further
revealed that the extracts prepared from the roots of A.
mexicana showed 1.6- to 2.4-fold higher efficacy than those
prepared from the leaves and stem. It establishes the hexane
root extract to be the most effective larvicide of all the extracts
tested resulting in LC50 and LC90 values of 91.331 and
156.684 ppm, respectively, after 24 of exposure (Table 2).
The extract proved to be 1.8-fold more toxic than hexane leaf
extracts (Table 3) and a 2.4-fold more toxic as against hexane
stem extracts (Table 4). Similarly, with an LC50 value of
158.839 ppm after 24 h of exposure, the petroleum ether root
extract exhibited 54.6 and 88.5 % increased efficacy as com-
pared to the petroleum ether stem and root extracts, respec-
tively. It is also worthy to note that the toxicity potential
increased after prolonged periods of exposure of the larvae
to the extracts; the LC50 values decreasing dramatically after
48 h of exposure with hexane root extract still being the most
effective larvicide (Fig. 2).
Behavioral observations on the larvae treated with all the
effective extracts of A. mexicana revealed interesting behav-
ioral symptoms. On immediate exposure to the extracts, all
the larvae exhibited a normal feeding and zigzag wriggling
motion till 510 min of exposure. Thereafter, the larvae
started showing signs of unnatural restlessness and excita-
tion. The wriggling speed increased conspicuously with
aggressive vertical movements rising repeatedly to the water
surface. Most interesting observation was the aggressive
self-biting of anal papillae with their mouth parts leading to
the formation of ring-shaped structures (Fig. 3). This aggres-
sive pattern persisted for 23 h after which the larvae became
sluggish, failing to reach the water surface, followed by high
levels of larval knockdown due to chronic paralysis. Mori-
bund or dead larvae were increasingly found from 4 to 8 h.
Observations on the morphological alterations of treated
larvae revealed that most organs had a normal structural
appearance except anal papillae (gills). Under light micro-
scope, both treated and control larvae showed similarities in
morphological architecture and cuticular sculpturing of the
head, thorax, and abdomen segments, and other organs such

Table 4 Larvicidal activity of the stem extracts of A. mexicana against early fourth instars of A. aegypti

Solvent Time of exposure (h) Larvicidal activity SE 2 (df) Regression coefficient

LC50 (ppm) LC90 (ppm)

Hexane 24 218.067 (182.979257.951) 452.843 (365.870633.991) 0.59 4.938 (4) 4.038

48 183.895 (152.114219.174) 403.041 (322.965569.905) 0.55 1.302 (4) 3.760
Petroleum Ether 4 245.595 (136.945492.640) 553.219 (329.4144,744.55) 0.56 5.901 (3) 3.634
48 214.438 (174.673265.704) 483.117 (369.787748.938) 0.56 3.809 (3) 3.633

Values in parentheses indicate the lower and upper 95 % fiducial limits

Parasitol Res

Fig. 1 Larvicidal efficacy of the

petroleum ether and hexane
extracts of A. mexicana against
A. aegypti after 24 h of exposure
to the extracts. PLE petroleum
ether leaf extracts, PSE
petroleum ether stem extracts,
PRE petroleum ether root
extracts, HLE hexane leaf
extracts, HSE hexane stem
extracts, HRE hexane root
extracts

as the eyes, antennae, mouth brushes, setae, saddle, siphon,
and ventral brushes. A distinct difference, however, was the
structural alteration of the anal gills observed in the extract-
treated larvae as compare to the control larvae. The observa-
tions clearly demonstrated destruction of anal papillae with
extensive damage and a shrunken cuticle of the internal mem-
brane whereas the external membrane was intact (Fig. 4).
Another remarkable observation was the demelanization of
the cuticle as compared to control (Fig. 5).
Discussion
Increasing documentation of negative environmental and
health impact of synthetic insecticides, and growing incidence
of resistance in the mosquitoes have necessitated the need for
the development of new strategies for mosquito control (Lima
et al. 2011). Keeping in view the development of insect
resistance to synthetic insecticides and the residue problems
in the environment, the current inclination is to explore plants
for certain phytochemicals that are safe for nontarget animals
and do not pose any residue problem but are still able to
suppress pest populations. During the last decade, various
studies on natural plant products against mosquito vectors
have renewed the interest in them as possible alternatives to
synthetic chemical insecticides. The botanical extracts from
the plant leaves, roots, seeds, flowers, and bark in their crude
form have been used as conventional insecticides for centu-
ries. In fact, many researchers have reported the effectiveness
of plant extracts or essential oils against mosquito larvae
(Rahuman et al. 2007; Rawani et al. 2009; Warikoo et al.
2011; Kumar et al. 2012b).
A. mexicana is an aggressive weed that grows in the tem-
perate region in waste lands, cultivating fields, and road sides.
In the traditional system of medicine, the whole plant of A.
mexicana is extensively used in the treatment of tumors, warts,

Fig. 2 Comparative larvicidal

efficacy of A. mexicana extracts
against A. aegypti after 24 and
48 h of exposure. PLE petroleum
ether leaf extracts, PSE
petroleum ether stem extracts,
PRE petroleum ether root
extracts, HLE hexane leaf
extracts, HSE hexane stem
extracts, HRE hexane root
extracts
 Parasitol Res

Fig. 3 Digital photomicrographs

of early fourth instar larvae of A.
aegypti; 1, 2, 3, and 4 different
time intervals of treatment with
effective extracts of A. mexicana.
The circled portions represent the
aggressive anal gill biting
behavior of extract-treated larvae
forming ring-shape

skin diseases, inflammations, rheumatism, jaundice, leprosy,
piles, warm infestations, and dysentery. Along with its pharma-
cological properties, a few researchers have reported its poten-
tial role in control of mosquito vectors (Sakthivadivel and
Thilagavathy 2008; Priya and Rao 2012). Keeping in view
limited research work and antimosquito potential of the plant,
the present investigations were carried out to assess the prospec-
tive use of A. mexicana in mosquito management programs.
Present investigations clearly revealed the efficacy of
extracts prepared from the three parts (leaf, stem, and root)
of A. mexicana against fourth instars of A. aegypti; though
only hexane and petroleum ether extracts, out of the five
solvents tested, could result in 80100 % mortality. Similar
results were reported by Kumar et al. (2012b) who tested
different parts of a weed, Parthenium hysterophorus,
extracted in five solvents against A. aegypti larvae and found
only hexane and petroleum ether extracts being effective
causing 100 % mortality. However, the acetone extracts
prepared from A. mexicana seeds have been proved to pos-
sess larvicidal potential against second instars of dengue
vector causing high mortality at test concentrations from 25
to 200 ppm (Sakthivadivel and Thilagavathy 2003).
Our results also confirmed the root extracts as the most
effective extracts with 1.6- to 2.4-fold higher efficacies as
compared to the leaf and stem extracts. These results are in
conformity with that of Kumar et al. (2012b) who also report-
ed the maximum efficacy of extracts prepared from the roots
of P. hysterophorus as compared to those prepared from stem
and leaves against dengue vector larvae. Larvicidal bioassays
with hexane root extracts of A. mexicana resulted in an LC50
and LC90 value of 91.331 and 156.684 ppm, respectively,
against early fourth instars of A. aegypti after 24 h of exposure.
The toxicity potential of the extract, however, increased by
1.1-fold on exposure to the extract for 48 h. In comparison,
hexane root extracts of P. hysterophorus have been found
moderately effective against fourth instars of A. aegypti with

Fig. 4 Micrographs of anal gills

of A. aegypti larvae. a Control
larva showing two pairs of
normal gills with sac-like
structure. b Extract-treated
larvae of A. aegypti showing four
anal gills with externally normal
appearance but internally
shrunken structure (arrows).
S-IAP shrunken internal anal
papillae, I-EAP intact external
anal papillae
Parasitol Res
Fig. 5 Extract-treated larvae of A. aegypti showing loss of pigmenta-
tion of cuticle as compared to control. DmC demelanized cuticle. 1
Control larva showing normal pigmentation of cuticle, 2 extract-treated
larva showing depigmentation of cuticle
LC50 and LC90 values of 432.38 and 1118.50 ppm, respec-
tively (Kumar et al. 2012b). This confirms the larvicidal
efficiency of A. mexicana against A. aegypti though further
investigations are needed for their use in the field.
Present studies also proved the larvicidal potential of pe-
troleum ether extracts with lowest LC50 value of 158.839 ppm
exhibited by root extracts. Earlier, Sakthivadivel and
Thilagavathy (2008) had reported the efficacy of petroleum
ether leaf and seed extracts of A. mexicana against all the three
species of mosquitoes with an LC50 value of less than 50 ppm.
Earlier, Raj Mohan and Ramaswamy (2007) found the leaf
extracts of the weed Aegeratina adenophora moderately ef-
fective against fourth instars of A. aegypti and Culex
quinquefasciatus reporting an LC50 value of 256.70 and
227.20 ppm, respectively, and suggested its use for mosquito
control in stagnant water bodies. The leaf and flower extracts
of the weed Lantana camara are also reported to exhibit
larvicidal activity against third and fourth instar larvae of A.
aegypti and C. quinquefasciatus with maximum mortality at
3.0 mg/mL (Sathish and Maneemegalai 2008). However, the
petroleum ether extract of Euphorbia tirucalli was found
significantly effective against the larvae of A. aegypti and C.
quinquefasciatus with LC50 values of 4.25 and 5.52 ppm
(Rahuman et al. 2007). Our studies establish the efficacy of
hexane and petroleum ether extracts prepared from different
parts of A. mexicana. Further, more larvicidal potential of
hexane extracts as compared to the petroleum ether extracts,
and too prepared from roots suggest that the presence of
certain chemical constituents in the hexane extract which
arrested the metabolic activities of the larvae.
The behavioral observations of the larvae treated with
selected extracts prepared from A. mexicana revealed exci-
tation, aggressive vertical, and horizontal movements in the
larvae. The symptoms being similar to those caused by
synthetic nerve poisons, i.e., excitation, convulsions, paral-
ysis, and death suggests that the extracts could possibly act
as cytolysin, affecting the neuromuscular coordination in
chemical synapses. Our findings corresponded to those of a
few earlier studies (Choochote et al. 2004, 2005; Dharmagadda
et al. 2005; Chaithong et al. 2006; Kumar et al. 2010); how-
ever, the toxic symptoms were observed at relatively different
time intervals. Our studies also showed a very interesting
behavior of aggressive anal biting in treated larvae. Earlier,
Becker et al. (2010) had reported the role of anal papillae in
regulation of electrolyte balance, which is required for the
sustainability of the life functions. This suggests that the cyto-
toxic effects of A. mexicana extracts led to the discharge of
fatal electrolyte from the anal region resulting in violent anal
biting.
The treated early fourth instars of A. aegypti also exhibited
structural alteration of anal papillae. This organ is involved in
the regulation of electrolyte levels. The internal structures of
the anal papillae in treated larvae showed remarkable shrink-
age, while the external features were normal in appearance.
These results are in accordance with the results of Chaithong
et al. (2006) and Kumar et al. (2010) who reported the re-
markable shrinkage in the internal structure of anal papillae in
the larvae of A. aegypti when treated with ethanolic extracts of
pepper, while most of the other organs of dead larvae had a
normal appearance. Earlier, Insun et al. (1999) also observed
the damaged anal papillae, with a shrunken cuticle border and
destroyed surface with loss of ridge-like reticulum in dead C.
quinquefasciatus larvae when treated with ethanolic extract of
Kaempferia galanga. It has been suggested that structural
deformation of anal papillae probably led to their dysfunction,
which may be intrinsically associated with the death of mos-
quito larvae (Chaithong et al. 2006). Earlier reports have
revealed that uptake and elimination of most ions in mosquito
larvae occur via the anal papillae, which was markedly re-
duced or lost in larvae with deformed or absence of papillae
(Garrett and Bradley 1984; Clements 1992). This suggests
that the morphological alterations of anal papillae could lead
to their dysfunction probably leading to an interruption of the
osmotic and ionic regulation.
Our investigations have established the potential of A.
mexicana against A. aegypti larvae and their benefits to de-
veloping new types of larvicides used for mosquito control.
However, the mechanism involved causing mortality of mos-
quito larvae is still unknown and needs to be studied further.
The variety of types and levels of active constituents in each
kind of extract may be responsible for the variability in their
potential against A. aegypti. Our investigations need further
exploration to find out and identify the bioactive constituent
involved and their mode of action present in A. mexicana
extracts with antimosquito potential. Further studies of the
formulated preparations for enhancing potency and stability,
toxicity, and effects on nontarget organisms and the environ-
ment, and field trials are needed to recommend A. mexicana as
an antimosquito product used to combat and protect from
mosquitoes in a control program. This undoubtedly will lead

to improved formulations with enhanced activity, which may
eventually become environmentally acceptable and replace
objectionable conventional insecticides for mosquito control.
Acknowledgments The authors are highly grateful to University Grants
Commission, New Delhi for providing research fellowship to carry out the
present investigations. Thanks are extended to Dr. Savithri Singh, Principal,
Acharya Narendra Dev College for providing infrastructure and research
facilities.
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