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Carpocapsa Rez., 2008
Carpocapsa Rez., 2008
Daro Fernandez
and Cristina Montagna3
1 Departamento
Argentina
de Qumica, Universidad Nacional del Comahue, Buenos Aires 1400, (8300) Neuquen,
Ruta 22, Km 1192, Allen, Ro Negro, Argentina
3 Escuela Superior de Salud y Ambiente, Universidad Nacional del Comahue, Buenos Aires 1400, (8300) Neuquen,
Argentina
2 INTA
Abstract
BACKGROUND: Azinphos-methyl is the main insecticide used to control codling moth on apple and pears in
Northern Patagonia. The aim of this study was to evaluate the toxicological and biochemical response of diapausing
larvae of codling moth in orchards subjected to different insecticide selection pressure.
RESULTS: Dosemortality assays with azinphos-methyl in diapausing larvae of Cydia pomonella L. showed
significant differences between the LD95 from a population collected in one untreated orchard (2.52 g moth1 )
compared with that in a laboratory-susceptible population (0.33 g moth1 ). Toxicity to azinphos-methyl in field
populations of diapausing larvae collected during 20032005 was evaluated by topical application of a discriminating
dose (2.5 g moth1 ) that was obtained from larvae collected in the untreated orchard (field reference strain).
Significantly lower mortality (37.7184.21%) was observed in three out of eight field populations compared with
that in the field reference strain. Most of the field populations showed higher esterase activity than that determined
in both the laboratory susceptible and the field reference strains. Moreover, there was a high association between
esterase activity and mortality (R2 = 0.64) among the field populations. On the other hand, a poor correlation
was observed between glutathione S-transferase activity and mortality (R2 = 0.33) among larvae collected from
different orchards.
CONCLUSIONS: All the field populations evaluated exhibited some degree of azinphos-methyl tolerance in
relation to the laboratory susceptible strain. Biochemical results demonstrated that esterases are at least one of
the principal mechanisms involved in tolerance to this insecticide.
2008 Society of Chemical Industry
Keywords: codling moth; Cydia pomonella; resistance; azinphos-methyl; esterases; glutathione S-transferases
1 INTRODUCTION
The codling moth Cydia pomonella L. (Lepidoptera: Tortricidae) is a major pest of apples
and pears throughout the world. Organophosphorus insecticides, particularly azinphos-methyl,
have been extensively used for control of codling
moth in the United States,1,2 Chile,3,4 the Czech
Republic5 and Australia.6 Consequently, resistance to this compound,2,3,4,7 9 phosalone5,10 and
parathion has occurred.11 Field-exposed populations
of codling moth have also developed resistance to
pyrethroids,2,12 insect growth inhibitors,10,13,14 insect
growth regulators3,10,15 and biological products.16
Moreover, field populations of codling moth resistant to azinphos-methyl have shown cross-resistance
Correspondence to: Cristina Montagna, Escuela Superior de Salud y Ambiente, Universidad Nacional del Comahue, Buenos Aires 1400, (8300)
Argentina
Neuquen,
E-mail: mmontagn@uncoma.edu.ar
(Received 3 August 2007; revised version received 3 December 2007; accepted 22 December 2007)
Published online 31 March 2008; DOI: 10.1002/ps.1582
et al.
Jimena Soleno
3 RESULTS
3.1 Toxicological assays on diapausing larvae
The azinphos-methyl responses of diapausing larvae
from both a laboratory (susceptible) and an untreated
orchard (Cinco Saltos) population are presented in
Fig. 1. The data from both doseresponse bioassays
showed a good fit to the probit model. The
LD95 (0.33 g larva1 ) of the laboratory population
was about eightfold lower than that observed in
the Cinco Saltos population (2.52 g larva1 ). The
slope from the laboratory population was very steep
(6.2 1.5) compared with that from the Cinco Saltos
population (3.1 0.3), and thus they exhibited only
fourfold difference at the LC50 level. The LC50
as well as the LC95 values from both populations
were significantly different according to their 95%
confidence limits. Because of the substantial difference
at the LC95 between these two strains, larvae from
Cinco Saltos were considered as the field reference
strain among the field populations. Indeed, it was
the only population that had not been recently
subjected to chemical or biological control (5 years)
at the time of these assays. Table 1 shows the
azinphos-methyl response of diapausing larva from
field populations subjected to different insecticide
selection pressure. Assays performed during 2003
showed that the mortality at the discriminating dose of
azinphos-methyl (2.5 g larva1 ) in diapausing larvae
collected at the locality of Cinco Saltos (96.55%) was
significantly higher than the mortality observed on
larvae collected in two orchards at Allen (38.33 and
84.21%). On the other hand, the mortality registered
on the population from Centenario (92.85%) did
not differ statistically from that exhibited by the
reference strain. Field populations from General Roca,
Guerrico and Valle Azul evaluated for azinphos-methyl
response during 2004 were found to be significantly
more susceptible (98.18, 98.21 and 98.24% mortality)
than that from Cinco Saltos (91.66%). Among
the populations collected during 2004, only larvae
from Vista Alegre (60.34%) showed significantly
lower mortality than larvae from Cinco Saltos at
the discriminating dose of azinphos-methyl. Larvae
collected during 2005 from the same orchard at
Vista Alegre showed not only significantly lower
mortality (37.71%) than that recorded in the Cinco
Saltos population (91.25%) but also significantly
lower mortality than that observed during the 2004
collection. Finally, larvae from Cinco Saltos collected
during 2003, 2004 and 2005 did not show statistical
differences in mortality.
3.2 Biochemical assays
Means from esterase and GST activities across
diapausing larvae from a laboratory susceptible
population and field populations are shown in Table 2.
Pest Manag Sci 64:964970 (2008)
DOI: 10.1002/ps
7
Laboratory
Cinco Saltos
PROBIT
80
60
40
20
% MORTALITY
95
3
0.1
1
DOSE (mg per larva1)
10
Azinphos-methyl
Populationa
Cinco Saltos (u)
Allen1 (t)
Allen2 (t)
Centenario (t)
General Roca (t)
Guerrico (t)
Valle Azul (o)
Villa Regina (t)
Vista Alegre (t)
Year of
collection
nb
Mortality (%)c
2003
2004
2005
2003
2003
2003
2004
2004
2004
2005
2004
2005
61
96
80
60
57
56
55
56
57
60
80
55
96.55
91.66
91.25
38.33
84.21
92.85
98.18
98.21
98.24
90.00
60.34
37.71
610.84
26.18
2.30
4.09
4.17
4.27
0.12
74.56
286.78
4 DISCUSSION
Two of eight field populations (25%) that were
collected during 20032005 displayed no significant
difference in mortality to azinphos-methyl to that
in the Cinco Saltos population. Furthermore, three
populations, including the one from an organic
orchard, registered significantly lower mortalities than
that of the field reference population. However,
larvae from the untreated orchard were found to
Table 2. Esterase and GST activities of diapausing larvae from a
laboratory susceptible and field populations
Laboratory
Cinco Saltos
Allen1
Allen2
Centenario
General Roca
Guerrico
Valle Azul
Villa Regina
Vista Alegre
Year of
collection
Esterase
GSTb
2004
2003
2004
2003
2003
2003
2004
2004
2004
2005
2004
2005
0.040 (0.0049)d
0.029 (0.0058)a
0.024 (0.0062)ab
0.14 (0.019)c
0.14 (0.026)c
0.018 (0.0072)b
0.019 (0.0069)b
0.045 (0.012)d
0.062 (0.0088)
0.18 (0.036)c
0.13 (0.037)c
0.20 (0.10)c
0.080 (0.017)a
0.077 (0.029)a
0.096 (0.033)a
0.24 (0.040)b
0.27 (0.074)b
0.27 (0.057)b
0.10 (0.045)a
0.077 (0.041)a
0.091 (0.033)a
nd
0.21 (0.034)b
nd
967
et al.
Jimena Soleno
ESTERASE ACTIVITY
(mmol min1 mg protein1)
R2 = 0.639
0.25
0.20
0.15
0.10
0.05
15
B
GST ACTIVITY
(mmol min1 mg protein1)
0.30
30
45
60
% MORTALITY
75
90
30
45
60
% MORTALITY
75
90
R2 = 0.326
0.25
0.20
0.15
0.10
0.05
15
pyrethroid detoxification. Azinphos-methyl susceptibility at that time was probably due to a prevailing
insecticide activation by MFO over detoxification
mechanisms.
Synergism of pyrethroids by organophosphorus
insecticides has been reported in resistant populations
ACKNOWLEDGEMENTS
The authors gratefully acknowledge Dr Benot
Sauphanor for valuable suggestions on a previous
version of the manuscript. This study was supported by
a grant from the Universidad Nacional del Comahue
Project I 940.
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