Iranian Journal of Entomology (2011) 1, 8-12

2011 IJE Efficacy of spinosad on stored-product pests 8

Efficacy of spinosad against three stored-product insect
pests

Maedeh Mollaie
1
, Hamzeh Izadi
1
and Hosein Dashti
2

1
Department of Plant Protection, Faculty of Agriculture, Vali-e-Asr University, Rafsanjan,
IRAN; and
2
Department of Agronomy and Plant Breeding, Faculty of Agriculture, Vali-e-
Asr University, Rafsanjan, IRAN


Summary
1. The insecticidal effects of spinosad (Tracer 24SC) on the adults of Tribolium
castaneum and the larvae of Ephestia kuehniella and Plodia interpunctella were
evaluated.
2. The mean mortality of T. castaneum was affected by both the concentration of and
the exposure time to spinosad.
3. Increasing spinosad concentration did not affect mortality of T. castaneum after 7
days. After 14 days, however, mortality was increased (up to about 90%) by an
increase in spinosad concentration (up to 1 mg/kg).
4. Spinosad at the concentrations 0.1, 0.25, 0.5, 0.75 and 1 mg/kg completely
suppressed larval survival of E. kuehniella.
5. At the concentration 1 mg/kg, spinosad suppressed larval survival and adult
emergence of P. interpunctella more than 90%.
6. Apart from the great insecticidal effects on stored-product pests, the efficacy of
spinosad may vary with the target pest species, its developmental stage, its specific
food source and the exposure time.

Key-words: Tribolium, Ephestia, Plodia, spinosad, stored products.

Introduction
The red flour beetle, Tribolium
castaneum (Herbst) (Coleoptera,
Tenebrionidae), Mediterranean flour moth,
Ephestia kuehniella Zeller (Lepidoptera,
Pyralidae), and Indian meal moth, Plodia
interpunctella (Hubner) (Lepidoptera,
Pyralidae), are the primary and most
destructive pests of stored nuts and grains;
they cause severe quantitative and
qualitative losses throughout the world
(Campbell and Arthur 2007).

Correspondence: Dr. Hamzeh Izadi,

Department of
Plant Protection, Faculty of Agriculture, Vali-e-Asr
University, Rafsanjan, IRAN.
E-mail: izadi@vru.ac.ir

Alleviation of these pests problems in
storage mostly relies on synthetic pesticides.
The intensive use of synthetic pesticides in
past few years, however, has not met the
modern criteria of integrated pest
management programs, leading to an
increasing interest for natural pesticides
derived from plants and microorganisms
(Sparks et al. 2001; Tedeschi et al. 2001).
Spinosad is an insecticide based on a
fermentation product of the soil
actinomycete bacterium, Saccharopolyspora
spinosa Mertz and Yao, which was
discovered during the 1980s. It is a broad-
spectrum insecticide with low mammalian
toxicity (Mertz and Yao 1990). Spinosad
Iranian Journal of Entomology (2011) 1, 8-12
2011 IJE Efficacy of spinosad on stored-product pests 9

exhibits a high degree of selective toxicity
towards many lepidopteran, dipteran and
thysanopteran insects. It, however, is less
toxic to many beneficial arthropods. This
biopesticide degrades quickly in exposure to
sunlight (Thompson et al. 2000), but it is
relatively stable in stored grains (Fang et al.
2002b; Flinn et al. 2004). Spinosad is toxic
to insects by ingestion and contact (Liu et al.
1999; Wanner et al. 2000) and has a unique
mode of action on the insect nervous system
at the nicotinic acetylcholine receptor and
gamma-amino butyric acid receptor sites
(Salgado 1998; Sparks et al. 2001). Poisoned
insects exhibit involuntary muscle
contractions and tremors, followed by hyper
excitation and paralysis (Salgado 1998).
An ideal grain protectant should combine
rapid mortality with long-term stability
(Arthur 1996). In storage facilities, in the
absence of light, spinosad remains stable for
a long period and thus, it can provide long-
term protection for stored grains (Fang et al.
2002a,b). This insecticide has been studied
in many countries, where it has been
recommended for the control of several
stored grain pests. The aim of the present
study was to assess the efficacy of spinosad
for the control of T. castaneum, E.
kuehniella and P. interpunctella.

Materials and Methods
Insecticide
A liquid formulation of spinosad
(Tracer 24SC, a commercial formulation
containing spinosyns A and D; Dow
AgroSciences, UK) was diluted in distilled
water to make solutions of different
concentrations for treatment.

Test insects
To initiate culturing, T. castaneum was
obtained from Ferdowsi University of
Mashhad (Mashhad, Iran), P. interpunctella
was obtained from Pistachio Research
Institute (Rafsanjan, Iran), and E. kuehniella
was obtained from Khorassan-Razavi
Research Center for Agriculture and natural
Resources (Mashhad, Iran). Tribolium
castaneum, E. kuehniella and P.
interpunctella were reared on a mixture of
whole wheat flour and yeast (20:1 w:w),
whole wheat flour and pistachio nut,
respectively. Colonies were maintained in a
climate chamber set at 252 C, 655% RH
and a photoperiod of L:D 16:8 h.

Bioassay
To assay efficacy of spinosad against all
the three pest species, Nayak et al. (2005)s
method with some modification was used.
To test spinosad on T. castaneum (the 1
st

experiment), 1 ml of different spinosad
concentrations were applied to 50 g wheat in
a 125-ml glass container, and shook for 2
min; this provided 0.1, 0.5, 1 and 2 mg
AI/kg. Untreated wheat (as control) received
1 ml of distilled water. Twenty five 1-7-day-
old adult T. castaneum were introduced into
each container.
To test spinosad on E. kuehniella (the 2
nd

experiment), 0.2 ml of different spinosad
concentrations were applied to 20 g wheat to
provide 0.1, 0.25, 0.5, 0.75 and 1 mg AI/kg.
After shaking, the treated wheat was
incubated at 60
o
C for 48 h in an oven,
ground into flour, and kept in 125-ml glass
containers. Thirty 1-24-h-old eggs of E.
kuehniella were then introduced in to each
container.
To test spinosad on P. interpunctella (the
3
rd
experiment), 0.2 ml of different spinosad
concentrations were applied to 10 g
pistachio nuts in 125-ml glass containers to
provide 0.1, 0.25, 0.5, 0.75 and 1 mg AI/kg.
After shaking, thirty 1-24-h-old eggs of P.
interpunctella were introduced in to each
container.
All the containers were then covered with
lids and incubated under the standard
environmental conditions (as used for
culturing). The experiments were replicated
three (the 1
st
experiment) or four (the 2
nd
and
3
rd
experiments) times. Mortality of adult T.
Iranian Journal of Entomology (2011) 1, 8-12
2011 IJE Efficacy of spinosad on stored-product pests 10

castaneum was recorded after 7 and 14 days.
The larval survival and adult emergence of
E. kuehniella and P. interpunctella were
recorded after 21 and 51 days, respectively.

Data analysis
The data on T. castaneum mortality were
analyzed using a two-way ANOVA to
determine the effects of exposure period and
concentration of spinosad. Because of the
lack of variance equality and residual
normality, the data on E. kuehniella were
analyzed using the nonparametric method
via Kruskal-Wallis H test. The other data
were subjected to a one-way ANOVA. Pair-
wise comparisons were performed using
Duncan's multiple range test. Due to
variance inequality in the data on adult
emergence of P. interpunctella, the groups
with different variances were analyzed
separately; the treatments within groups
were then compared using Students t-test.
All statistical analyses were completed in
Minitab14 and MSTAT-C.

Results
The effects of spinosad on the mortality of T.
castaneum adults
There were significant differences in the
mean mortality of T. castaneum between
exposure times (F
1,16
= 77.32, P < 0.001)
and between concentrations (F
4,16
= 112.94,
P < 0.001). In addition, the interaction
between exposure time and concentration
was significant (F
4,16
= 7.28, P < 0.01). The
adult survival was significantly different
between concentrations in both the exposure
times (7 days: F
4,8
= 24.28, P < 0.001; 14
days: F
4,8
= 134.62, P < 0.001). Increasing
spinosad concentration did not affect
mortality after 7 days. After 14 days,
however, mortality was increased by an
increase in spinosad concentration up to 1
mg/kg (Table 1).



Table 1. The effects of spinosad on the mortality of
Tribolium castaneum adults
1
Means within columns followed by different letters
are significantly different (P < 0.01; Duncan's
multiple range test).

The effects of spinosad on the larval survival
and adult emergence of E. kuehniella
There were significant effects of spinosad
on the mean larval survival (H
5
= 22.76, P <
0.001) and on the mean adult emergence (H
5
= 22.76, P < 0.001) of E. kuehniella. In
control, about 72% of the eggs developed to
larvae after 21 days, and about 58% of the
eggs emerged to adults after 51 days. On the
contrary, spinosad at the concentrations 0.1,
0.25, 0.5, 0.75 and 1 mg/kg completely
suppressed larval survival and adult
emergence (Table 2).

Table 2. The effects of spinosad on the larval
survival and adult emergence of Ephestia kuehniella
Spinosad Performance of Ephestia kuehniella
concentration
(mg/kg)
Larval survival
1
(%; mean SE)
Adult emergence
2
(%; mean SE)
0.00 72.50 5.16 58.33 6.16
0.10 0 0
1
Survived larvae from treated eggs after 21 days
2
Emerged adults from treated eggs after 51 days

The effects of spinosad on the larval survival
and adult emergence of P. interpunctella
Treatment with spinosad was
significantly influenced the larval survival
(F
5,18
= 38.38, P < 0.001) and adult
emergence (F
3,12
= 138.89, P < 0.001) of P.
interpunctella. In control, 65% of the eggs
developed to larvae after 21 days, and about
56% of the eggs emerged to adults after 51
days. At the concentration 1 mg/kg,
spinosad suppressed larval survival and
adult emergence more than 90% (Tale 3).
Spinosad Mortality (%; mean SE)
concentration Exposure time to spinosad
(mg/kg) 7 days 14 days
0.0 (control) 8.0 4.0 a
1
8.0 4.0 a
0.1 46.7 3.5 b 61.3 3.5 b
0.5 54.0 2.0 b 74.0 2.0 c
1.0 60.0 6.1 b 89.3 2.7 d
2.0 61.3 1.3 b 97.3 1.3 d
Iranian Journal of Entomology (2011) 1, 8-12
2011 IJE Efficacy of spinosad on stored-product pests 11

Table 3. The effects of spinosad on the larval
survival and adult emergence of Plodia
interpunctella
Spinosad Performance Plodia interpunctella
concentration
(mg/kg)
Larval survival
1
(%; mean SE)
Adult emergence
2
(%; mean SE)
0.00 65.00 4.81 a
3
56.67 2.36 a
4
0.10 60.00 4.91 a 40.83 6.87 ab

0.25 32.50 6.99 b 25.00 6.17 bc
0.50 11.67 2.15 c 10.00 2.36 c
0.75 12.50 0.83 c 11.67 0.96 c
1.00 5.00 2.15 c 5.00 2.15 c
1
Survived larvae from treated eggs after 21 days
2
Emerged adults from treated eggs after 51 days
3
Means within columns followed by different letters
are significantly different (P < 0.01; Duncan's
multiple range test).
4
Means comparisons using Students t-test (see
Table 4 for more detailed analyses)

Discussion
Spinosad belongs to a new class of
insecticides, which act through rapid
excitation of the insect nervous system (Liu
et al. 1999; Wanner et al. 2000). The present
study revealed that spinosad can be
effectively used against T. castaneum, E.
kuehniella and P. interpunctella. The
efficacy of spinosad, however, may vary by
factors such as the target pest species, its
developmental stage, its specific food source
(commodity) and the exposure time. Similar
effects of spinosad have been reported on
other stored-product insect pests
(Mutambuki et al. 2003; Chintzoglou et al.
2008).
In the present study, mortality of T.
castaneum adults increased with an increase
in spinosad concentration at 14-day but not
at 7-day exposure time. In addition, the
mortality of T. castaneum was increased as a
function of exposure time. Other studies has
been also reported the slow-acting feature of
this insecticide (Foster et al. 1996; Wanner
et al. 2000; Huang et al. 2007).
It was clear that E. kuehniella larvae were
more susceptible to spinosad compared with
P. interpunctella larvae. At the highest
spinosad concentration tested (1 mg/kg)
only a 95% suppression of larval survival
and adult emergence of P. interpunctella
was achieved. On the contrary, excellent
control of E. kuehniella was achieved at the
lowest concentration of spinosad tested;
spinosad at 0.1 mg/kg completely
suppressed larval survival (100% mortality)
of E. kuehniella, such that no larvae
developed to pupa. Differences in toxicity of
spinosad observed in the present study might
be due to the different commodity used as
pest food (Fang et al. 2002a; Chintzoglou et
al. 2008). As found by other studies (Fang et
al. 2002a; Huang et al. 2007; Huang and
Subramanyam 2007), the susceptibility of P.
interpunctella larvae to spinosad was dose
dependent.

Table 4. The mean comparisons of the adult
emergence of Plodia interpunctella using Students t-
test
Spinosad Difference in adult emergence (%)
concentrations Spinosad concentrations (mg/kg)
(mg/kg) 0.100 0.250
0.00 15.84
NS 1
31.67 *
0.10 15.83
NS

0.25 15.83
NS

0.50 30.83 * 15.00
NS

0.75 29.16 * 13.33
NS

1.00 35.83 * 20.00
NS

1
NS and * denote non-significance (P > 0.05) and
significance (P < 0.05) of the difference,
respectively.

Suppression of the subsequent
generations is one of the basic
characteristics of a successful grain
protectant (Arthur 1996). Spinosad is
capable of giving long-term protection
without the loss of efficacy (Fang et al.
2002b; Athanassiou et al. 2011). However,
there are some indications of reproduction
even with high parental mortality (Fang et
al. 2002b). Here, spinosad at 0.1-1 mg/kg
provided an effective suppression of T.
castaneum, E. kuehniella and P.
interpunctella.
Iranian Journal of Entomology (2011) 1, 8-12
2011 IJE Efficacy of spinosad on stored-product pests 12

In summary, it was indicated that even as
a slow acting insecticide, spinosad has a
great potential for control of the stored-grain
pests. However, depending on the species
and commodities the different
concentrations of spinosad are required to
get satisfactory results.

Acknowledgements
This work was supported by a Research
Vice Presidency of Vali-e-Asr University
grant to Hamzeh Izadi.

References
Arthur FH (1996) Grain protectants: current status
and prospects for the future. Journal of Stored
Product Research 39, 293-302.
Athanassiou CG, Arthur FH, Kavallieratos NG,
Throne JE (2011) Efficacy of spinosad and
methoprene, applied alone or in combination,
against six stored-product insect species. Journal
of Pesticide Science 84, 61-67.
Campbell JF, Arthur FH (2007) Ecological
implications for post harvest integrated pest
management of grain and grain-based products.
Ecologically Based Integrated Pest Management
(ed. by Koul O, Cuperus GW), CAB
International, Wallingford, pp. 406-431.
Chintzoglou G, Athanassiou CG, Arthur FH (2008)
Insecticidal effect of spinosad dust, in
combination with diatomaceous earth, against two
stored-grain beetle species. Journal of Stored
Product Research 44, 347-353.
Fang L, Subramanyam B, Arthur FH (2002a)
Effectiveness of spinosad on four classes of wheat
against five stored-product insects. Journal of
Economic Entomology 95, 640-650.
Fang L, Subramanyam B, Dolder S (2002b)
Persistence and efficacy of spinosad residues in
farm stored wheat. Journal of Economic
Entomology 95, 1102-1109.
Flinn PW, Subramanyam B, Arthur FH (2004)
Comparison of aeration and spinosad for
suppressing insects in stored wheat. Journal of
Economic Entomology 97, 1465-1473.
Foster RN, Reuter KCL, Black E, Kabla BF, Flora
EA (1996) Laboratory evaluation of dose and
manner of pickup of spinosad on the migratory
grasshopper. Arthropod Management Tests 21,
416-417.
Huang F, Subramanyam B (2007) Effectiveness of
spinosad against seven major stored-grain insects
on corn. Insect Science 14, 225-230.
Huang F, Subramanyam B, Hou X (2007) Efficacy of
spinosad against eight stored-product insect
species on hard white winter wheat. Biopesticides
International 3, 117-125.
Liu TX, Sparks AN, Hendrix WH, Yue B (1999)
Effects of SpinTor (spinosad) on cabbage looper
(Lepidoptera: Noctuidae): toxicity and persistence
of leaf residue on cabbage under field and
laboratory conditions. Journal of Economic
Entomology 92, 1266-1273.
Mertz FP, Yao RC (1990) Saccharopolyspora
spinosa sp. nov. isolated from soil collected in a
sugar mill rum still. International Journal of
Systematic Bacteriology 40, 3439.
Mutambuki K, Ngatia CM, Mbugua JN, Likhayo P
(2003) Evaluation of the efficacy of spinosad dust
against major storage pests. Proceedings of the
Eight International Conference on Stored-
Product Protection. 22-26 July 2002, York, UK.
CAB International Wallingford, Oxon, pp 888-
891.
Nayak MK, Daglish GJ, Byrne VS (2005)
Effectiveness of spinosad as a grain protectant
against resistant beetle and psocid pests of stored
grain in Australia. Journal of Stored Product
Research 41, 455-467.
Salgado VL (1998) Studies on the mode of action of
spinosad: insect symptoms and physiological
correlates. Pesticide Biochemistry and Physiology
60, 91-102.
Sparks TC, Crouse GD, Durst G (2001) Natural
products as insecticides: the biology,
biochemistry, and quantitative structure-activity
relationships of spinosyns and spinosoids. Pest
Management Science 57, 896-905.
Tedeschi R, Alma A, Tavella L (2001) Side-effects
of three neem (Azadirachta indica A. Juss)
products on the predator Macrolophus caliginosus
Wagner (Het., Miridae). Journal of Applied
Entomology 125, 397402.
Thompson GD, Dutton R, Sparks TC (2000)
Spinosad-a case study: an example from natural
products discovery program. Pest Management
Science 56, 696702.
Wanner KW, Helson BV, Harris BJ (2000)
Laboratory and field evaluation of spinosad
against the gypsy moth, Lymantria dispar. Pest
Management Science 56, 855-860.




Received: 25 May 2011
Accepted: 21 September 2011
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