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Effects of piperonyl butoxide (PBO) as a synergist with deltamethrin on five

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J. Commun. Dis. 39 (3) 2007 : 159-163

Efficacy of Piperonyl Butoxide (PBO) as a Synergist with

Deltamethrin on Five Species of Mosquitoes
Vijayan V A* +, Sathish Kumar, B Y*, Ganesh K N*, Urmila, J.*
Fakoorziba M R* and Makkapati A K**
(Received Aug., 2006)

ABSTRACT
Development of insecticide resistance has been a challenging problem for a
long time and new solutions are yet to emerge. In this regard, the use of
synergist with the insecticide is thought to play a key role in reducing the
resistance levels. Present study demonstrates the efficacy of PBO with
deltamethrin against the field collected mosquito larvae of five species of
Aedes, Anopheles and Culex from in and around Mysore.
KEY WORDS : Deltamethrin, piperonyl butoxide, resistance, synergist, Aedes,
Anopheles, Culex.

INTRODUCTION mammalian toxicity due to its high

Widespread occurrence of resistance to
insecticides is a serious threat for the control
and management of many important insect
pests.1 Resistance to insecticides has been
noted in Culex quinquefasciatus2 Bencheik et
al., 1998; Anopheles stephensi3 Anopheles
gambiae4, Anopheles albimanus5 and Aedes
aegypti.6 In this context, integrated pest
management draws attention, as it is
important to save the environment, man and
his livestock from chemical pollution.
Rotations, mosaics and mixtures have all
been proposed as resistance management
tools Of late, there has been mounting interest
in the use of synergist to reduce the resistance
incidence by combined applications.
Presently, deltamethrin is considered to be
one of the most promising synthetic
pyrethroids, due to its efficacy and its
photostable nature with very low
vulnerability to enzymatic degradation.10 It is
well established that PBO is an effective
synergist for natural pyrethrins and synthetic
pyrethroids.11, 12 As it is a well known inhibitor
of microsomal monooxygenase, which are
involved in the metabolism and
detoxification of virtually all insecticides.13,14
The present study was undertaken to
evaluate the efficacy of deltamethrin with
PBO against field collected mosquito larvae.
Aedes aegypti, Anopheles culicifacies, Anopheles
stephensi, Culex tritaeniorhynchus and Culex
quinquefasciatus were the species tested.
Deltamethrin selected line of Aedes aegypti
for 26 generations also was employed for
the study.
MATERIALS AND METHODS
Fourth instar larvae of five species collected
in and around Mysore from the paddy fields,
ground pools, sewage tanks, tree holes,

* Department of Studies in Zoology, Manasagangothri, Mysore-570 006.

** Bayer (India) Limited, Bayer House, Central Avenue, Powai, Mumbai 400 076
160 Vijayan V. A et al.

artificial containers like tins, broken pots, deltamethrin + PBO on 19 to 26 generation.

tyres etc., were employed. Technical grade
deltamethrin with 98.8% purity and the
synergist, piperonyl butoxide [M/s. Nexus
(Endura) make] were obtained through
Bayer (India) Ltd. Mumbai. Different
concentrations of deltamethrin were
prepared and serial dilutions of deltamethrin
+ PBO in the ratio 1:6 were made. Tests were
kept simultaneously for deltamethrin and
deltamethrin + PBO along with the control.
Early fourth instar larvae were used for
bioassay, in accordance with the standard
WHO method for mosquito larvae (WHO,
1981). The larvae, in groups of 25, were
treated for 24 hrs with 1ml ethanolic solutions
of insecticides in 249ml of water in a 500ml
glass beaker. Control larvae were exposed to
ethanol alone and mortality was corrected
with Abbott's formula.15 In each treatment the
insecticide concentration was adjusted so as
to generate about 50% mortality. The larvae
surviving after 24hrs were thoroughly
washed and reared to adults. The larvae of
the subsequent generations were again tested
the same way and further selected with the
approximate LC 50 dose. The selection
experiments were continued for 26
generations Aedes aegypti with deltamethrin.
The selected line was treated with
The mortality data were subjected to
regression analysis of probit-mortality on log
dosage and their LC50, slope and
heterogeneity (÷2) about the linear regression
line were computed16 (Finney, 1971) and
percent suppression10 was calculated.
RESULTS
The dose response values and resistance
suppression data are given in Table 1. Of the
five species tested with deltamethrin, Aedes
aegypti had shown very low LC50 value of
0.00021mg/l, followed by Culex
quinquefasciatus, Culex tritaeniorhynchus,
Anopheles culicifacies and Anopheles stephensi
with 0.00078mg/l, 0.00152mg/l,
0.00204mg/l and 0.00418mg/l respectively.
With deltamethrin-PBO combination, Aedes
aegypti has again shown least LC50 value of
0.00001mg/l and Anopheles culicifacies has the
maximum of 0.00046mg/l. When resistance
ratio and percent suppression in tolerance
was taken in to consideration, Aedes aegypti
showed the highest difference of 21.00 in
terms of LC50 values and 96.3% suppression in
tolerance. The lowest difference of 4.00 in
terms of LC50 values and 75.0% suppression in
tolerance was noticed in case of Culex
tritaeniorhynchus.

Table 1. Larval LC50 (ppm), resistance ratio and percent suppression of 5 species of
mosquitoes at Mysore tested with Deltamethrin and Deltamethrin + PBO.

Species Deltamethrin Deltamethrin + PBO Percent

LC50 Slope ± SE ÷ (df) 2 LC50 Slope ± SE ÷ 2(df) Resistance suppression
(ppm) Heterogeneity (ppm) Heterogeneity ratio

A. aegypti 0.00021 1.2384 ± 0.1405 3.29(4) 0.00001 1.4285± 0.2380 5.93(3) 21.00 95.23
An. culicifacies 0.00204 2.0048+0.2601 3.95(3) 0.00035 2.3441+0.3558 9.05(3) 5.83 82.84
An. stephensi 0.00418 1.4163±0.1716 3.57(3) 0.00023 1.7514±0.1879 3.99(3) 18.17 94.52
Cx.
tritaeniorhynchus 0.00152 1.0265±0.1850 10.96(4) 0.00038 1.4784±0.3777 49.78(4) 4.00 75.00
Cx.
quinquefasciatus 0.00078 4.2513±0.5372 7.2(3) 0.00019 5.4305±0.5732 2.4(3) 4.10 75.70
 Efficacy of piperonyl butoxide 161

Results of the selection experiments on deltamethrin against all the six species. The
Aedes aegypti with deltamethrin and the toxicity of deltamethrin against a laboratory
combined effect with PBO on these selected strain of Culex quinquefasciatus from India as
lines are given in Table 2. The results with reported by Rajvanshi et al.,18 and Thomas et
deltamethrin indicate significant increase in al.,10 was found to be more, than that observed
the LC50 values in descendent line compare to in the present investigation. Similarly,
the F1. An increase of 333.83 folds tolerance deltamethrin is known to have a very high
was noticed in the 16th generation in terms of larvicidal activity against various species of
LC50 values17 and 2269.5 fold tolerance in the Aedes as well.19, 20, 18, 21, 22
26th generation. The LC50 value of 26th It is clear that deltamethrin in
generation larvae, when treated with combination with PBO brought down the
deltamethrin + PBO was 0.065017mg/l. That tolerance level in all the species tested
is, the resistance ratio of F26 deltamethrin and (Table1). In line with this, earlier Thomas et
Deltamethrin + PBO was 9.42 in terms of LC50 al., and Kumar et al., have shown 17.3% and
value with 89.38% suppression in tolerance. 6.7% suppression in resistance in laboratory
DISSCUSSION strains of Culex quinquefasciatus and Aedes
aegypti respectively, when treated with
Incidences in resistance to all classes of
deltamethrin+PBO(1:5).10, 20 Casida et al., and
insecticides have made vector control more
Kumar et al have shown that piperonyl
and more difficult. Employing synergists
butoxide is an effective synergist with natural
with insecticides could be promising in
pyrethrins and synthetic pyrethroids due to
controlling the insects as many are major
its ability to inhibit the pivotal detoxifying
vectors. The present data also have reiterated
enzymes the monooxgenases.11, 10 It was found
the superior larvicidal activity of
to suppress the deltamethrin tolerance level
in An. stephensi, Ae. aegypti and Cx.
Table 2. Differential susceptibility of quinquefasciatus in Delhi, India.10 Rodriguez et
Ae.aegypti to Deltamethrin and
al., also have reported a significant reduction
Deltamethrin + PBO during selection
in pyrethroids tolerance level when applied
experiment.
with PBO against Cx. quinquefasciatus from
LC 50 (ppm) Resistance Percent Cui dal de La Habana.23 Similarly, Vulule et
ratio suppression
Generations Deltamethrin Deltamethrin al., (1999) too, have observed a reduction in
+ PBO pyrethroid tolerance level with PBO against
F1 0.000210 0.000010 21.00 95.23 An. gambiae in Kenya.24
F17 0.148620 - - -
Present observations on the larval
F18 0.153330 - - - selection experiments with deltamethrin is in
F19 0.256104 0.018942 13.52 92.60 line with the earliers findings in Ae. aegypti
F20 0.368487 0.042820 8.600 88.37 and Cx. quinquefasciatus respectively.20, 10 Jinfu
F21 0.383560 0.044395 8.630 88.42 had also recorded an increase in resistance to
F22 0.390921 0.046947 8.320 87.99 deltamethrin in the larvae of different strains
F23 0.483590 0.052701 9.700 89.10 of Cx. pipiens pallens after field application of
F24 0.488947 0.055620 8.790 88.62 deltamethrin in Southeast China. 2 5 A
F25 0.599367 0.057747 10.38 90.36 significant finding in our study was the effect
F26 0.612776 0.065017 9.420 89.38
of the synergist, PBO on the selected lines of
Ae. aegypti. When F 26 larvae were treated with
162 Vijayan V. A et al.
deltamethrin + PBO combination, the builtup
resistance, based on LC50 values have come
down from 2918 to 310. So, from the present
investigations, it is clear that PBO could be
employed with deltamethrin in endemic
areas, where continuous application is
needed. These results are significant, as
deltamethrin is being employed by the Public
Health Department of Karnataka and
suggest PBO as an effective synergist with
deltamethrin against Aedes, Anopheles and
Culex species.
ACKNOWLEDGEMENT
Authors are grateful to the Chairman,
Department of Studies in Zoology,
University of Mysore, Manasagangothri,
Mysore for the facilities. The financial
assistance provided by M/s. Aventis Crop
Science India (now M/s. Bayer Crop Science
Ltd.) and M/s. Nexuss (Endura) for the
supply of PBO is gratefully acknowledged.
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