1
Ti le:
1
Moni o ing esis ance o
Helico e pa a mige a
o di e en insec icides used in co on
2
in Spain.
3
4
Au ho s: Ca los A illa, José E. González-Zamo a
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A ilia ions: Depa men o Ciencias Ag o o es ales, Uni e si y o Se ille. Ca e e a
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de U e a, km 1. E-41012-Se ille (Spain)
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Co esponding au ho : José E. González-Zamo a
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zamo [email protected]
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Phone: +34-954 486 459
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Fax: +34-954 46 436
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Published in: C op P o ec ion 29 (2010) 100–103
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doi:10.1016/j.c op o.2009.09.007
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2
Abs ac
1
Helico e pa a mige a
is he key pes o co on in Spain, esul ing in many
2
insec icide ea men s agains i . The esis ance s a us o
Helico e pa a mige a
o
3
di e en insec icides cu en ly used in co on was e alua ed in Spain in wo di e en
4
seasons, 1999 and 2004. Fou popula ions we e es ed in o al, wo in each season.
5
Toxicological bioassays we e conduc ed in he labo a o y, and pe o med on hi d
6
ins a la ae by opical applica ion o he insec icides. LD50’s we e es ima ed by
7
p obi analysis and esis ance ac o s (RF) we e calcula ed a he LD50 le el. Fou
8
insec icides we e e alua ed, bu only endosul an eached a mode a e esis ance le el
9
(RF=11.4), and he o he s (me homyl, chlo py i os and lambda-cyhalo h in) showed
10
low esis ance (RF be ween 1.9 and 6.0). Such esul s indica e he gene ally low
11
esis ance o
H. a mige a
o mos o he insec icides used agains his pes in co on
12
in Spain. Possible explana ions o his si ua ion a e discussed.
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Key wo ds: insec icide esis ance, co on,
Helico e pa a mige a
, endosul an,
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me homyl, chlo py i os, lambda-cyhalo h in.
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3
1. In oduc ion
1
Co on is an impo an c op in he Guadalqui i i e alley (Andalucía, sou he n
2
Spain), especially in he p o inces o Se ille, Có doba and Cádiz. One o i s main
3
economic cos s comes om insec icide ea men s agains pes s like
Helico e pa
4
a mige a
Hübne (Lepidop e a: Noc uidae), he key pes o co on in sou he n Spain
5
(Du án, 1999).
H. a mige a
can se e ely educe co on yields, mainly because i
6
eeds on lowe buds, lowe s and on he boll. I s con ol is based mainly on an IPM
7
schedule, which in ol es es ima ing popula ions o he pes and i s na u al enemies:
8
p edado y bugs (He e op e a) and lacewings (Neu op e a), and ecommending
9
insec icides only when
H. a mige a
popula ions su pass a h eshold and na u al
10
enemies a e sca ce (Du án, 1999; 2003).
11
12
The con ol o his pes in Spanish co on may equi e se e al applica ions
13
h ough he season ( he IPM co on is sp ayed wi h insec icides be ween 2 and 9
14
imes, ange ob ained om 23 seasons, Du án (1999; 2003)), mainly o endosul an
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and me homyl (2-6 ea men s), complemen ed wi h o he p oduc s including
16
py e h oids (e.g. lambda-cyhalo h in), o ganophospha es (e.g. chlo py i os) o o he
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ca bama es (e.g. hiodica b) (Du án, 2003). The con inuous use o hese p oduc s
18
can lead o he de elopmen o esis an popula ions o
H. a mige a
, and in he 2003
19
season impo an con ol ailu es o
H. a mige a
we e obse ed in co on ields. The
20
ailu e epo s in ol ed mainly he Se ille p o ince and he wo p oduc s ha we e
21
mos used agains his ca e pilla , endosul an and me homyl. The e a e many epo s
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o he esis ance o
H. a mige a
(and o he Helio hinae) o insec icides a ound he
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wo ld, especially o py e h oids, bu also o o he g oups o insec icides, like
24
4
cyclodienes (such as endosul an), ca bama es (such as me homyl and hiodica b)
1
and o ganophospha es (such as clo py iphos) (McCa e y, 1999; Bues e al., 2005;
2
Ahmad e al., 2007; Pie an onio e al., 2007; Ugu lu and Gu kan, 2007; Saleem e
3
al., 2008).
4
5
Co on is a good c op in which o s udy he p esence o esis ance o insec icides
6
due o he high numbe o ea men s ha may be applied (Deguine e al., 2008),
7
bu un il now no speci ic s udy has been done in his c op in Spain, e en hough,
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wi h G eece and Bulga ia, i is one o he ew coun ies in he Eu opean Union whe e
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co on is cul i a ed. On he o he hand, ou s udy complemen s o he s ca ied ou in
10
Spain o analyze he s a us o esis ance in
H. a mige a
in di e en c ops (To es-Vila
11
e al., 2002a,b). This s udy co e ed wo seasons: he 1999 season, in which he
12
baseline esis ance o he insec icides ha a e mos equen ly used agains
H.
13
a mige a
in co on was es ablished wi h a labo a o y s ain, and compa ed wi h a
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ield popula ion, and he 2004 season, in which he e olu ion o suscep ibili y o he
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same insec icides o e he pe iod o 5 yea s, and con ol ailu es o
H. a mige a
in
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co on epo ed in he p e ious 2003 season, we e analysed.
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2. Ma e ial and me hods
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The
H. a mige a
popula ions used in his s udy we e ob ained om co on ields
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in Se ille and Có doba (Andalucía, sou he n Spain) om la ae collec ed in di e en
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plo s. In 1999, he labo a o y s ain was c ea ed om a numbe o pupae om a
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colony ha had been main ained in he Uni e si y o Có doba by P o esso E. Va gas-
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Osuna since 1996, wi h pe iodical ein oduc ions o indi iduals om co on ields,
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5
and wi h no con ac wi h any kind o insec icide. The 1999 ield s ain was ob ained
1
in June by collec ing la ae om di e en co on ields om he p o ince o Se ille.
2
In he 2004 season, wo popula ions we e collec ed, he i s in June (p io o
3
applica ion o insec icides agains
H. a mige a
in ha season) om la ae om
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di e en co on ields in he p o ince o Se ille in which con ol ailu es o
H.
5
a mige a
had been obse ed in he p e ious 2003 season, and he o he in
6
Sep embe , om he same o nea by co on plo s.
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8
La ae we e b ough o he labo a o y and ea ed ollowing he me hodology o
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Poi ou and Bues (1974), a 26±1 ºC and 55±10 %RH wi h a 16:8(L:D) pho ope iod.
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Adul s we e ea ed in e ical cylind ical ubes o 11 cm in diame e and 26 cm in
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heigh , cons uc ed om labo a o y d ying pape and co e ed wi h pape . Th ee o
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i e pai s o adul s we e placed in each ube o ma ing and o iposi ion in he
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condi ions desc ibed abo e, and we e ed wi h a 10% honey solu ion. Eggs we e
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collec ed in co on bundles and placed in con aine s. A e ha ching, la ae we e ed
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wi h a semi-syn he ic die (Poi ou and Bues, 1974) and ea ed in he condi ions
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desc ibed abo e. Thi d ins a la ae we e ans e ed o indi idual cages o 4 cm in
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diame e and 2 cm in heigh , wi h a po ion o die , o insec icide bioassays.
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Labo a o y es ing o he insec icides was ca ied ou wi h he i s o second
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gene a ion o la ae ea ed in he labo a o y. Toxicological bioassays we e pe o med
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on hi d ins a la ae using he s anda d opical applica ion p ocedu e as
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ecommended by he En omological Socie y o Ame ica (Anonymous, 1970) and
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ollowing he guidelines o Robe son and P eisle (1992). Fou p oduc s we e used:
24

6
endosul an (Thiodan-35, 350 g l-1 a.i. om Baye C opScience), me homyl (Bonsul,
1
200 g l-1 a.i. om Baye C opScience), chlo py i os (Cuspide-48, 480 g l-1 a.i. om
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Come cial Química Massó), and lambda-cyhalo h in (Ka a e King, 25 g l-1 a.i. om
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Syngen a). Fo each insec icide, se ial dilu ions in ace one [o 0.075% humec an
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(Ag idexa, om Baye C opScience) in wa e o lambda-cyhalo h in] we e p epa ed
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so ha each was one-hal o he p e ious dose. A d op o solu ion (2 µl) was applied
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wi h a 0.5–10 µl mic opipe e o he ho acic do sum o each hi d ins a la ae.
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Con ols o each eplica e we e ea ed wi h ace one (o wa e wi h Ag idexa) alone.
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The cages we e closed wi h a lid en ila ed by a hole co e ed wi h mesh, and le in
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he same ea ing condi ions desc ibed abo e. Fo each s ain, a leas 240 la ae
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we e ea ed pe insec icide, usually wi h 4 eplica es o 10 la ae a each o 5 o
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mo e insec icide concen a ions. A e 48 hou s mo ali y was assessed: la ae we e
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conside ed dead i hey we e desicca ed, o unable o mo e when dis u bed wi h a
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b ush.
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Dose-mo ali y eg essions, LD50’s and hei iducial limi s we e es ima ed by
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p obi analysis (Finney, 1971) using he POLO Plus 1.0 so wa e (LeO a So wa e,
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Be keley, CA, USA). Resis ance ac o s (RF) we e es ima ed a he LD50 le el as RF=
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LD50 collec ed s ain/ LD50 e e ence s ain (ei he he 1999 labo a o y s ain o he
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s ains collec ed in he ield in 1999 o June 2004). The insec icide esis ance le el
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was classi ied acco ding o he c i e ia epo ed in To es-Vila e al. (2002a,b):
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suscep ibili y (RF=1), low esis ance (RF=2–10), mode a e esis ance (RF=11–30),
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high esis ance (RF=31–100), and e y high esis ance (RF=>100).
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3. Resul s and Discussion
1
The esis ance in
Helico e pa a mige a
o di e en insec icides ha a e usually
2
applied in Spain on co on was es ed in wo seasons (Table 1). In all cases, he
-
3
a io o he slope was signi ican (P<0.05), so ha a signi ican dose- esponse line
4
was ob ained.
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Bo h in e -season and in a-season compa isons can be made wi h he da a. In
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he in e -season compa ison, he esul s showed ha he LD50 o all p oduc s es ed
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had inc eased a he end o he 2004 season (in Sep embe ), when compa ed
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agains he LD50 o he 1999 labo a o y s ain, wi h he iducial limi s no
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o e lapping, excep in chlo py i os. This inc emen was mos ob ious wi h
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endosul an, which eached he ca ego y o mode a e esis ance (RF=11.4), whe eas
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he o he p oduc s we e placed in he ca ego y o low esis ance (RF ange be ween
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1.9 and 6.0). Compa ison o he da a om Sep embe 2004 wi h he ield popula ion
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o 1999 p oduced RFs ha anged om 1.1 o 3.1, i.e., om suscep ibili y o low
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esis ance, bu only endosul an had iducial limi s ha did no o e lap, indica ing an
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inc emen in i s LD50 in his pe iod.
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18
The second kind o compa ison is in a-season. The only compa ison ha is
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meaning ul is o he yea 2004, when ield popula ions we e collec ed a he
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beginning and a he end o he season. The Sep embe popula ion showed
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suscep ibili y/low esis ance (RF be ween 0.6 and 1.7) when compa ed wi h he June
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popula ion, bu no inc emen o he LD50 was de ec ed, wi h he iducial limi s
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o e lapping o all he insec icides.
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8
1
The esul s ob ained in his wo k a e simila o hose ob ained in a s udy by
2
To es-Vila e al. (2002a,b) du ing 1995 o 1999, which included he same
3
insec icides es ed he e, bu in di e en c ops (excluding co on). They ound he
4
LD50 o endosul an o be be ween 0.93 and 6.70 µg/la a, which includes ou esul .
5
In e ms o he RF, he alue ob ained in his s udy was highe (RF=11.4, included in
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he mode a e esis ance ca ego y), bu his is p obably because i was compa ed
7
wi h a mo e suscep ible labo a o y s ain. I he less suscep ible s ain o To es-Vila
8
e al. (2002a) had been used ins ead, hen he RF would be 3 (included in he low
9
esis ance ca ego y).
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11
The o he insec icides es ed in his s udy (i.e., me homyl, chlo py i os and
12
lambda-cyhalo h in) ollowed he same pa e n as endosul an: hei LD50 is included
13
in he ange o alues epo ed in To es-Vila e al. (2002a,b), and wi h RF alues
14
ha a e always wi hin he suscep ibili y o low esis ance ca ego y.
15
16
A clea inc emen o he LD50 has been de ec ed in mos o he insec icides es ed
17
be ween 1999 and 2004, compa ed wi h he labo a o y s ain. Only a small change
18
was obse ed in he LD50 in he pe iod om 1999 o 2004 in he ield s ains (only
19
signi ica i e in he case o endosul an), and no signi ican inc emen was de ec ed in
20
he 2004 season. O e all, ou esul s showed li le o no di e ence om he esul s
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ob ained in he gene al s udy ca ied ou om 1995 o 1999 by To es-Vila e al.
22
(2002a,b) in Spain, e en hough he s udy p esen ed he e included wo seasons,
23
1999 and 2004, ha we e e y sepa a e in ime. Such esul s indica e a gene al
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9
scena io o low esis ance o
H. a mige a
o almos all he p oduc s es ed in Spain,
1
excep o some py e h oids (cype me h in, lambda-cyhalo h in and del ame h in),
2
o which To es-Vila e al. (2002b) ound high o e y high le els o esis ance,
3
al hough only in 4 ou o 111 insec icide–s ain combina ions es ed, in ol ing 2 ou
4
o 35
H. a mige a
s ains.
5
6
The le el o esis ance in
H. a mige a
o endosul an in ou s udy can be
7
conside ed as low o mode a e. This is simila o he si ua ion in o he co on-
8
g owing egions such as hose in Tu key (Ugu lu and Gu kan, 2007) and India
9
(K an hi e al., 2002; Su yawanshi e al., 2008). In e ms o o he insec icides, like
10
me homyl, se e al o ganophospha es, and especially py e h oids (such as
11
cype me h in, del ame h in, lambda-cyhalo h in, and o he s), he e a e many s udies
12
om di e en coun ies epo ing high le els o esis ance in his species (K an hi e
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al., 2002; Pie an onio e al., 2007; Ugu lu and Gu kan, 2007; Su yawanshi e al.,
14
2008). S udies o insec icide esis ance in
Helico e pa/Helio his
in Eu ope ha e no
15
been limi ed o Spain, and in F ance he e is also e idence o signi ican le els o
16
esis ance o py e h oids (del ame h in), and mode a e esis ance o me homyl in
H.
17
a mige a
, bu no esis ance has been de ec ed agains endosul an
(Bues e al.,
18
2005).
19
20
The de elopmen o insec icide esis ance is p ima ily he esul o he selec ion
21
p essu e exe ed on sp ayed popula ions, which inc eases he equency o esis an
22
indi iduals. Howe e , he co on c op in Spain is sp ayed wi h insec icides agains
H.
23
a mige a
om 2 o 6 imes pe season (Du án, 2003), which does no help o explain
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16
Table 1. Toxici y o opically applied insec icides, cu en ly used in co on in
1
Spain, o di e en s ains o
Helico e pa a mige a
la ae (F1 and F2, hi d ins a ), in
2
wo yea s.
3
Insec icide
Yea –S ain
In e cep
Slope±S.E.
LD50
(µg/la a)
95%
iducial
limi s
RFa
Endosul an
1999–Labo a o y
6.31
2.19±0.34
0.25
0.18–0.32
--
1999–Field
5.05
1.81±0.26
0.93
0.72–1.28
3.7
2004–June
4.54
2.11±0.27
1.64
1.30–2.05
6.6
2004–Sep embe
4.10
1.97±0.33
2.86
1.44–5.03
11.4
Me homyl
1999–Labo a o y
6.57
1.43±0.28
0.08
0.05–0.12
--
1999–Field
6.14
1.92±0.27
0.26
0.19–0.33
3.3
2004–June
5.46
1.01±0.16
0.35
0.22–0.50
4.4
2004–Sep embe
5.47
1.48±0.22
0.48
0.19–0.90
6.0
Chlo py i os
1999–Labo a o y
7.83
3.06±0.40
0.12
0.09–0.14
--
1999–Field
7.16
3.21±0.46
0.21
0.17–0.25
1.8
2004–June
5.88
2.21±0.28
0.40
0.32–0.50
3.3
2004–Sep embe
6.41
2.22±0.20
0.23
0.11–0.41
1.9
Lambda-Cyhalo h in
1999–Labo a o y
7.88
1.63±0.28
0.02
0.00–0.03
--
1999–Field
7.12
1.40±0.25
0.03
0.02–0.04
1.5
2004–June
6.87
1.64±0.19
0.07
0.06–0.09
3.5
2004–Sep embe
7.08
1.85±0.27
0.08
0.05–0.12
4.0
In all s ain–insec icide combina ions he p obi dose- esponse lines we e signi ican (
- a io
4
es ,
P
<0.05).
5
a Resis ance ac o (RF) es ima ed as RF= LD50 speci ic s ain/LD50 1999 labo a o y s ain.
6
7