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122

Bioefficacy of Skeeter Abate and Spintor on Aanopheles gambiae
and Aedes aegypti Mosquitoes from insecticide resistance areas in
Lagos and Oyo States, Nigeria

*Kemabonta K. A., Anikwe, J.C and Adaezeobiora, I. B
Department oI Zoology, Faculty oI Science, University oI Lagos, Akoka, Lagos, Nigeria.
* E mail oI corresponding author: kennykemabontayahoo.com

ABSTRACT
Introduction: The development oI resistance in mosquitoes to the insecticide used against them as larvicides and
adulticides is an important issue to be considered as diIIiculty with control would be encountered due to the
emergence oI insecticide -resistant mosquitoes. Resistance to Organophosphates and Pyrethroids, widely used to
control Mosquitoes is widespread in Nigeria . Objectives: This study thereIore evaluates the eIIect oI insecticide
resistance on the larvicidal activity oI Skeeter Abate and Spintor a biopesticide, on Anopheles gambiae and Aedes
aegvpti. Methods: Mosquito larvae were collected Irom Iive populations where insecticide resistance have been
reported in Lagos and Oyo States in Nigeria. They were evaluated at a temperature oI 27+ 2
o
c and R.H. oI 70+ 10
and a photoperiod oI 12:12 (LD). Insecticide resistance was conIirmed in the areas by adult susceptibility test using
World Health Organization standard procedures. The larval bioassay was conducted at diIIerent concentrations oI
Skeeter Abate and Spintor insecticides. Results: There is a positive correlation between resistance to permethrin in
adult mosquitoes and the concentration oI Spintor and Skeeter Abate required to eIIect 99 mortality in the larval
samples. All Aedes aegvpti species collected Irom the study sites were 100 susceptible to permethrin. Resistance to
permethrin in Anopheles gambiae were low in three sites (Yaba, Ikorodu and Iwo Road) while resistance is
suspected in two sites (Lekki and Ojoo). Spintor and Skeeter Abate were eIIective in the control oI the Anopheles
gambiae and Aedes aegvpti larvae. Conclusion: Insecticide susceptibility tests and resistance surveillance should be
conducted beIore embarking on mosquito control programmes.
Keywords: Insecticide resistance Mosquitoes, Aedes aegvpti, Anopheles gambiae, biopesticide, Spintor and Skeeter
Abate.

INTRODUCTION
Vector-borne diseases are an increasing cause oI death and suIIering worldwide. OI all the disease-spreading
arthropods, mosquitoes, the greatest public health problem (WHO, 2005), cause physical, mental and health eIIects
to man and animals. WHO declared the mosquito 'public enemy number one because they are responsible Ior the
transmission oI various dreadIul diseases such as malaria, Iilariasis, yellow Iever, dengue Iever and several types oI
encephalitis (WHO, 1996, Sachs & Malaney, 2002; Boutayeb, 2006).

Malaria pathogens are transmitted to humans by the bite oI Iemale Anopheles mosquitoes. About 50 oI Nigerians
suIIer Irom at least one episode oI malaria each year. Yellow Iever is a viral disease which is mainly transmitted to
humans by Aedes aegvpti. The WHO estimates that the disease aIIects an estimated 200,000 persons, causing an
estimated 30,000 deaths (WHO, 2006) yearly.
The emergence oI mosquito species resistant to insecticides widely used in malaria and dengue control has the
potential to impact severely on the control oI these disease vectors. The Insecticide Resistance Action Committee
(IRAC) suggests the use oI Insecticide Resistance Management (IRM) to help reduce the negative eIIects oI
resistance. The objective oI IRM is to prevent or delay the evolution oI resistance to insecticides or to help regain
susceptibility in already resistant insect populations (IRAC, 2005). A National Research Council report (NRC, 1986)
on strategies and tactics Ior pesticide resistance management described insecticide susceptibility as a resource in
resistance management.
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Skeeter Abate is an organo-phosphorus insecticide highly eIIective against mosquito larvae and other aquatic
insects, while its toxicity to Iish, birds, mammals and humans is very low. Its low toxicity to non-target organisms
and low eIIective dosage make Skeeter Abate an appropriate larvicide in many situations (WHO, 1997). Presently,
Skeeter Abate is being used in larvicidal programmes in Lagos State, Nigeria.
Spintor is a naturally derived biorational insecticide which is highly toxic to Lepidoptera, Diptera and some
Coleoptera and has a unique mode oI action involving the postsynaptic nicotinic acetylcholine and GABA receptors
resulting in tremors, paralysis, and death (Salgado, 1998). It has a very low mammalian toxicity and a Iavourable
environmental proIile with low persistence and low toxicity to a number oI predatory insects (Miles & Dutton,
2000).
Insecticide resistance in adult populations oI An. gambiae and Ae. aegvpti has been reported in many studies
(Shousha, 1948, Oduola et al., 2010, Ranson et al., 2010, Kamgang et al., 2011, Oduola et al., 2012). However, the
eIIects oI this resistance on the success oI larviciding as a part oI IVM have not been well documented. This study
thereIore evaluates the eIIect oI insecticide resistance on the larvicidal activity oI Skeeter Abate and Spintor on An.
gambiae and Ae. aegvpti mosquitoes. Such knowledge is essential in deIining Iuture control strategies against this
medically important mosquito. There has been no report on the susceptibility status oI anopheles mosquitoes Irom
Lekki, Lagos. Our Iinding thus provides this inIormation.

MATERIALS AND METHODS
Mosquito sampling
Anopheles gambiae and Aedes aegvpti larvae were collected Irom six sites (Ikorodu, Yaba, Lekki, and Ojoo (in
Lagos State) and Iwo road (in Oyo State) where insecticide resistance have been earlier suspected (Fig. 1). Larvae
were collected Irom diIIerent breeding habitats ranging Irom shallow, well, gutters, tyres, to standing water between
May and October, 2012. They were scooped into bowls, sieved, put in small buckets and washed with clean water
and were taken to the insectary Ior rearing to adult stage.

Rearing of larvae to adult stage in the insectary
The larval samples collected in the Iield were reared in the insectary oI the Vector Control Research Unit, National
Institute oI Medical Research (NIMR), Lagos, Nigeria. They were Ied with mashed biscuit oI low Iat (12.30g) plus
yeast capsules (7.59g) and maintained under a standard insectary conditions (temperatures between 27-29
o
c and
relative humidity between 80-97). The emerged adults were transIerred into cages using an aspirator and
maintained on 10 sucrose solution Ior 24 hours prior to Iurther tests.
Anopheles gambiae s.s. third larval instar (Kisumu strain) were Irom the Molecular Entomology and Vector Control
Research Laboratory, Public Health Division, Nigerian Institute oI Medical Research, Lagos. The mosquito colony
established in 2002 under the WHO/MIM TDR Project A30026 and has not been previously exposed to any Iorm
oI insecticide. The colony was maintained at a temperature oI 27+ 2
o
c and R.H. oI 70+ 10 and a photoperiod oI 12
12 (LD).

Preparation of stock solutions and test concentrations.
Stock solutions were prepared by weighing 5.0mg, 10.0mg, 15.0mg, 20.0mg and 25.0 mg oI the Spintor wettable
powder into Iive Eva water bottles labelled A
SPD
to E
SPD
respectively, 1000ml oI distilled water was poured into each
bottle and then screwed. The screwed bottles were shaken vigorously to dissolve the chemical inside. Stock solutions
were used as Ireshly prepared stock solution.
Skeeter Abate pellets were ground into powder using a mortar and pestle. Stock solutions were prepared by weighing
0.5mg, 1.0mg, 1.5mg, 2.0mg and 2.5mg oI the ground Skeeter Abate powder into Iive Eva water bottles labelled
A
SKE
to E
SKE
respectively, 1000ml oI distilled water was poured in each bottle and then screwed. The screwed bottles
were shaken vigorously to dissolve the chemical inside. Stock solutions were used as Ireshly prepared stock solution.



Insecticide bioassay
Bioassays were carried out on non-blood Ied 2 to 3 day old Iemale mosquitoes that emerged Irom the larvae
collected Irom the Iield. Standard procedures were Iollowed Ior susceptibility tests using WHO test kits and
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insecticide-impregnated papers obtained Irom Vector Control Research Unit, NIMR (WHO, 1998). One test paper
impregnated with permethrin at 1.0 concentration was used to assess the presence oI resistance. The mosquitoes
Irom each location were tested with 3 replicates oI 20 to 25 samples per area; these were then exposed Ior 60 minutes
to each oI the insecticides or to untreated paper (control). The number oI mosquitoes knocked down was recorded
every ten minutes oI exposure aIter which the insects were maintained Ior 24 hours on 10 sucrose solution and
mortality oI the insect was recorded. The results were analyzed to obtain the LC
50
by using computerised probit
analysis (Raymond, 1985). The WHO (1998) criteria Ior determining resistance or susceptibility were applied: 98 to
100 mortality indicates susceptibility, 80 to 97 mortality requires conIirmation oI resistance. While 80
mortality suggests resistance.

Larval bioassay
The susceptibility oI larvae oI Anopheles gambiae and Aedes aegvpti to Spintor and Skeeter Abate was tested in the
laboratory using a methodology adapted Irom the Elliot larval test (WHO, 1997). Groups oI 20 larvae oI the late
third and early Iourth instars were transIerred by means oI droppers to small test disposable cups. 100ml oI the
diIIerent concentrations oI the stock solutions were measured out using a 100ml measuring cylinder and poured into
the diIIerent cups containing the larvae. Three replicates oI 20 larvae were assigned to each treatment. A cup
containing 100ml oI water was used as control. Mortality responses were recorded aIter 24 hours. A larva was
classiIied as dead iI it did not move when gently touched with the point oI a pipette and those incapable oI rising to
the surIace or not showing the characteristic diving reaction when the water is disturbed were also counted as dead
larvae.

RESULTS
Insecticide susceptibility varied considerably in Anopheles gambiae mosquitoes collected Irom the Iive sites. The 24
hours mortality rate recorded in Lekki, Yaba, Ikorodu, Ojoo and Iwo road were 86.3, 98, 100, 93.4 and
100, respectively (Table 1). Resistance is suspected in adult A gambiae in lekki and Ojoo areas. On the other
hand, 100 mortality was Iound in A. Aegvpti adults in all the locations. Thus, all adult Aedes mosquitoes are still
susceptible to permetrin.

Larvicidal efficacy of spintor on anopheles gambiae larvae.
Larvicidal eIIicacy oI Spintor in the Iive study sites is as shown in Figure 2. Generally, the larvae mortality increased
with increase in concentration oI the larvicide. The 24hour mortality oI wild strains oI An. gambiae larvae collected
Irom Ikorodu, Yaba, Lekki, Iwo road and Ojoo ranged Irom 53-100, 96-100, 18-98, 48-96 and 8-91
respectively. Larvae collected Irom Yaba were the most susceptible with 100 mortality at 10mg/l concentrations
and above. This was Iollowed by larvae collected Irom Ikorodu, Iwo road, Lekki and Ojoo (Fig.2) areas.

Larvicidal efficacy of skeeter abate on anopheles gambiae larvae.
The 24hour mortality oI wild strains oI An. gambiae larvae collected Irom Ikorodu was 100 while Ior Ojoo, Yaba,
Iwo and Lekki areas, it ranged Irom 95-100, 70-96, 48-96 and 5-100 respectively (Fig. 3). Larvae
collected Irom Ikorodu were the most susceptible with 100 mortality at all the concentrations. This was Iollowed
by larvae collected Irom Ojoo, Yaba, Iwo road and Lekki (Fig. 3). Aedes aegvpti larvae collected Irom the study
sites were 100 susceptible to Skeeter Abate.
The regression analyses (Figs 4 -13) show positive correlation between larval mortality and the concentration oI
Spintor and Skeeter Abate respectively. The larvae mortality increased with increase in concentration oI both
insecticides.
A comparison oI the eIIicacies oI the two larvicides in the selected sites is shown in Table 2. The relative
perIormance in terms oI 50 and 95 mortality across all samples were comparatively equal. However, Skeeter
abate perIormed non signiIicantly in Iwo road as compared with Spintor Irom the same site.

Evaluation of the correlation between inscticide resistance and larvicide resistance
EIIect oI insecticide resistance was compared across all populations. Anopheles mosquitoes collected Irom Lekki
and Ojoo sites showed higher LC
99
values as compared with the rest oI the populations and, these were sites where
resistance to permethrin were detected. Susceptible sites showed lower LC
99
values (Table 3).
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DISCUSSION
The success oI malaria vector control programs could be daunted by the menace oI insecticide resistance. Resistance
oI mosquitoes to insecticides in Nigeria has been extensively studied (Awolola et al., 2003; 2005; Djouaka et al.,
2008; Oduola et al., 2010; 2012). There have been Iew reports in Lagos and Oyo States on the susceptibility status oI
Aedes mosquitoes to Permethrin impregnated papers using WHO standard methods. The 24 hour mortality results oI
this study shows that Spintor and Skeeter Abate were eIIective against Aedes aegvpti larvae and adults producing
100 mortality.
In Lagos State, resistance oI anopheles mosquitoes to various classes oI insecticides is becoming more alarming
(Oduola et al., 2010). Awolola et al., 2003 reported the resistance oI anopheles mosquitoes to pyrethroid insecticides
while Djouaka et al.,( 2008) reported the resistance to permethrin in Ojo in Oyo State. From our studies, resistance is
suspected in some parts oI Lagos State (Lekki ) and not Iound in the other parts studied. There has been no report
on the susceptibility status oI Anopheles mosquitoes Irom Lekki thus, our Iinding could be said to provide this
inIormation.
There has been a Iew studies on the eIIectiveness oI Spintor and Skeeter Abate on Anopheles gambiae larvae.
However, a lot oI works have studied the eIIect oI Spintor and Skeeter Abate on Aedes aegvpti mosquito larvae. Our
results agree with the study carried out by Darriet et al., (2005) which showed that Spintor had a lethal action on
susceptible and resistant specimens oI An. gambiae. The result oI the study carried out by Marina et al. (2011)
showed that Skeeter Abate granules and a suspension concentrate Iormulation oI Spintor were both highly eIIective
larvicides against Ae. aegvpti and Ae. albopictus. Moderate adult insecticide resistance values were recorded Irom
two populations out oI Iive (Ojo in Oyo State and Lekki in Lagos State). There have been a Iew reports on larval and
adult resistance to insecticides which has varied signiIicantly. Tikar et al., (2011) reported some level oI
susceptibility to Skeeter Abate and malathion in the larvae oI Anopheles mosquitoes, Rodriguez et al., (2007) on
the other hand evaluated levels oI resistance in Aedes aegvpti and reported that in larval bioassays, Skeeter Abate
resistance was high with high resistance oI the adults to pyrethroids.
In conclusion, resistance to insecticides could pose a major challenge to vector control programs and completely
render control programs ineIIective thus insecticide susceptibility tests and resistance surveillance is an essential step
in resistance management and should be conducted beIore undertaking mosquito control programmes. Spintor and
Skeeter Abate are very eIIective in the control oI mosquito larvae in areas where insecticide resistance is suspected.



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Figure 1: Map showing the larval collection sites


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Table 1: Resistance Status of Anopheles and Aedes adult mosquitoes from the selected sites

Site GPS coordinate 24hours
mortality
value
Susceptibility
status (WHO,
1998)
24hours
mortality
value
Susceptibility
status (WHO,
1998)
Longitude Latitude Aedes Anopheles
Lekki 3
0
27.983` 6
0
25.746` 100 Susceptible 86.3 Resistance
suspected
Yaba 3
0
22.275` 6
0
30.987` 100 Susceptible 98 Susceptible
Ikorodu 3
0
30.644` 6
0
38.013` 100 Susceptible 100 Susceptible
Ojoo 3
0
55.017` 7
0
27.812` 100 Susceptible 93.4 Resistance
suspected
Iwo road 3
0
56.497` 7
0
24.042` 100 Susceptible 100 Susceptible




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Figure 2: Larvicidal efficacy of Spintor in the five study sites after 24 hours.





Figure 3: Larvicidal efficacy of Skeeter Abate (Skeeter Abate) in the five study sites after 24 hours

m
o
r
t
a
|
|
t
y
Concentrat|on mg]|
SlnCSAu ?aba
SlnCSAu lwo road
SlnCSAu C[oo
SlnCSAu lkorodu
SlnCSAu Lekkl

m
o
r
t
a
|
|
t
y
concentrat|on mg]|
SkL11L8 ?aba
SkL11L8 lwo road
SkL11L8 C[oo
SkL11L8 lkorodu
SkL11L8 Lekkl
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Figure 4: Regressions analysis for Spintor using Anopheles mosquitoes collected from Ikorodu


Figure 5: Regressions analysis for Spintor using Anopheles mosquitoes collected from Yaba


Figure 6: Regressions analysis for Spintor using Anopheles mosquitoes collected from Lekki




y = 2.28x + 31.6
8 = 0.8038

m
o
r
t
a
|
|
t
y

Concentrat|on mg]|
SINCSAD IkCkCDU
y = 0.16x + 96.8
8 = 0.3

m
o
r
t
a
|
|
t
y
Concentrat|on mg]|
SINCSAD A8A
y = 3.78x + 14.7
8 = 0.8194

m
o
r
t
a
|
|
t
y
Concentrat|on mg]|
SINCSAD LLkkI
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Figure 7: Regressions analysis for Spintor using Anopheles mosquitoes collected from Iwo Road


Figure 8: Regressions analysis for Spintor using Anopheles mosquitoes collected from Ojoo


Figure 9: Regressions analysis for Skeeter abate using Anopheles mosquitoes collected from
Ikorodu

y = 2.42x + 39.9
8 = 0.8388

m
o
r
t
a
|
|
t
y
Concentrat|on mg]|
SINCSAD IWC kCAD
y = 3.32x + 8.2
8 = 0.7866

m
o
r
t
a
|
|
t
y
Concentrat|on mg]|
SINCSAD CICC
y = 100
8 = #n/A

m
o
r
t
a
|
|
t
y

Concentrat|on mg]|
SkL11Lk IkCkCDU
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Figure 10: Regressions analysis for Skeeter abate using Anopheles mosquitoes collected from Yaba


Figure 11: Regressions analysis for Skeeter abate using Anopheles mosquitoes collected from Lekki


Figure 12: Regressions analysis for Skeeter abate using Anopheles mosquitoes collected from Iwo
road


y = 18x + 61
8 = 0.73

m
o
r
t
a
|
|
t
y
Concentrat|on mg]|
SkL11Lk A8A
y = 31.4x - 24.3
8 = 0.938

m
o
r
t
a
|
|
t
y
Concentrat|on mg]|
SkL11Lk LLkkI
y = 32.4x + 24.4
8 = 0.8181

m
o
r
t
a
|
|
|
t
y

Concentrat|on mg]|
SkL11Lk IWC kCAD
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Figure 13: Regressions analysis for Skeeter abate using Anopheles mosquitoes collected from Ojoo


y = 4.8x + 89.4
8 = 0.6316

m
o
r
t
a
|
|
t
y
Concentrat|on mg]|
SkL11Lk CICC
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Table 2: Performance of Spintor and Skeeter Abate on Anopheles mosquitoes larval samples from
the selected sites in Lagos and Oyo States


X
2
values are Ior the test oI Iit oI the long time probit model used to estimate the MT
50
and
MT
95
; nsdeviations not signiIicant, *deviations signiIicant, P0.05






LARVICIDE SAMPLE
SITE
TOTAL
NUMBER
EXPOSED (N)
MORTALITY
MT
50
(95 CI)
MORTALITY
MT
95
(95 CI)
X
2

SPINOSAD YABA (60) 65.88(38.61-91.99) 81.14(44.97-123.37) 3.73*
SKETTER YABA (60) 52.27(32.50-85.42) 67.76(56.33-79.21) 7.98*
SPINOSAD IWO - ROAD (60) 48.21(30.58-67.25) 57.73(35.01-86.51) 14.68*
SKETTER IWO - ROAD (60) 25.46(13.92-35.43) 47.65(34.52-100.28) 2.01
ns

SPINOSAD OJOO (60) 57.44(45.51-88.46) 66.96(58.66-98.11) 43.6*
SKETTER OJOO (60) 62.41(45.24-80.81) 88.90(83.68-95.20) 7.88*
SPINOSAD IKORODU (60) 49.89(46.44-5432) 69.96(63.66-79.45) 38.2*
SKETTER IKORODU (60) 67.76(56.33-80.21) 95.1(88.89-102.83) 9.11*
SPINOSAD LEKKI (60) 46.87(39.14-63.12) 62.51(56.94-70.45) 5.02*
SKETTER LEKKI (60) 45.00(29.02-70.20) 75.88(48.61-101.99) 21.01*
Journal oI Biology, Agriculture and Healthcare www.iiste.org
ISSN 2224-3208 (Paper) ISSN 2225-093X (Online)
Vol.3, No.3, 2013

135


Table 3: Table showing the Susceptibility status of Anopheles mosquitoes and there effect on
Spinosad and Temephos.

LARVICIDE LC
99
(mg/l)
SUSCEPTIBILITY
VALUE (Adults)
SUSCEPTIBILITY
STATUS (Using
W.H.O criteria)
SAMPLE
LOCATION
SPINOSAD 0.9 mg/l 98 Susceptible Yaba
SKEETER 2.0 mg/l
SPINOSAD 20 mg/l 100 Susceptible Ikorodu
SKEETER 1.0 mg/l
SPINOSAD 25 mg/l 86.3 Resistance suspected Lekki
SKEETER 2.5 mg/l
SPINOSAD 25mg/l 93.4 Resistance
Suspected
Ojoo
SKEETER 2.0 mg/l
SPINOSAD 23.6 mg/l 100 Susceptible Iwo road
SKEETER 2.25 mg/l





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