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Laboratory induced bifenthrin resistance selection in Oxycarenus hyalinipennis (Costa) (Hemiptera: Lygaeidae): Stability, cross-resistance, dominance and effects on biological fitness
Journal Pre-proof Laboratory induced bifenthrin resistance selection in Oxycarenus hyalinipennis (Costa) (Hemiptera: Lygaeidae): Stability, cross-resistance, dominance and effects on biological fitness Ansa Banazeer, Sarfraz Ali Shad, Muhammad Babar Shahzad Afzal PII:
S0261-2194(20)30040-5
DOI:
https://doi.org/10.1016/j.cropro.2020.105107
Reference:
JCRP 105107
To appear in:
Crop Protection
Received Date: 28 December 2019 Revised Date:
2 February 2020
Accepted Date: 6 February 2020
Please cite this article as: Banazeer, A., Shad, S.A., Shahzad Afzal, M.B., Laboratory induced bifenthrin resistance selection in Oxycarenus hyalinipennis (Costa) (Hemiptera: Lygaeidae): Stability, cross-resistance, dominance and effects on biological fitness, Crop Protection (2020), doi: https:// doi.org/10.1016/j.cropro.2020.105107. This is a PDF file of an article that has undergone enhancements after acceptance, such as the addition of a cover page and metadata, and formatting for readability, but it is not yet the definitive version of record. This version will undergo additional copyediting, typesetting and review before it is published in its final form, but we are providing this version to give early visibility of the article. Please note that, during the production process, errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain. © 2020 Published by Elsevier Ltd.
Credit Author Statement AB conceived the study. MBSA and SAS designed the study. AB collected the insects and carried out the research work. SAS supervised the laboratory work. MBSA provided technical guidelines to process the raw data. AB analyzed the data. AB and MBSA wrote the manuscript. SAS read and approved the written manuscript.
1.4 1.2 Relative fitness
50 40 30 20 10
1 0.8 0.6 0.4 0.2
0
0 G1
G3
G5
G7
G9
G13
UNSEL
Bifen-Sel
Generations bioassayed with bifenthrin
Cross1
Populations
0.84 Degree of dominance
Selection of bifenthrin resistance
60
0.82 0.8 0.78 0.76 0.74 0.72 0.7 Cross1
Cross2
Hybrid populations
Cross2
1
Laboratory
induced
bifenthrin
resistance
selection
in
Oxycarenus
2
hyalinipennis (Costa) (Hemiptera: Lygaeidae): stability, cross-resistance,
3
dominance and effects on biological fitness
4
Ansa Banazeer,a Sarfraz Ali Shad,a* Muhammad Babar Shahzad Afzala,b**
5
a
6
Zakariya University, Multan, Punjab, Pakistan
7
b
8
Running Title: Bifenthrin resistance selection and comparison of fitness costs
9
To whom correspondence should be addressed:
Department of Entomology, Faculty of Agricultural Sciences and Technology, Bahauddin
Citrus Research Institute, Sargodha, Punjab, Pakistan
10
*Dr. Sarfraz Ali Shad
11
Department of Entomology, Faculty of Agricultural Sciences and Technology, Bahauddin
12
Zakariya University, Multan, Pakistan
13
Email: [email protected]
14
**Dr. Muhammad Babar Shahzad Afzal
15
Department of Entomology, Faculty of Agricultural Sciences and Technology, Bahauddin
16
Zakariya University, Multan, Pakistan and Citrus Research Institute, Sargodha, Punjab, Pakistan
17
Email: [email protected]
18
ABSTRACT
19
The dusky cotton bug, Oxycarenus hyalinipennis (Costa) (Hemiptera: Lygaeidae) is a serious
20
pest of cotton and damages cotton seed by reducing the oil content. In Pakistan, O. hyalinipennis
21
is managed by using various insecticides and has developed resistance to several insecticides. In
22
this study, O. hyalinipennis was selected with bifenthrin for 12 generations (G1 to G12) and
23
developed a 55.64-fold resistance when compared with an unselected population (Unsel Pop).
24
Bifenthrin resistance declined from 55.64 to 24.93-fold when selected population was removed
25
from bifenthrin selection pressure for four generations (G13 to G16). Bifenthrin selected (Bifen-
26
Sel) population showed a very low cross-resistance to profenofos (2.82-fold), deltamethrin (2.35-
27
fold) and acetamiprid (2.21-fold) when compared with a field population (Field Pop). The
28
bifenthrin resistance was incomplelety dominanat and autosomal. The relative fitness (Rf) of
29
Bifen-Sel population was 0.58 along with significant decreases in average nymphal survival,
30
fecundity, egg hatchability, intrinsic rate of population increase (rm), net reproductive rate (Ro)
1
31
and biotic potential (Bp). The Rf of Cross1 and Cross2 was 1.07 and 1.22, respectively. The high
32
fitness costs, instable resistance and a very low cross-resistance with other insecticides might be
33
useful in slowing down the evolution of bifenthrin resistance by implementing an insecticide
34
rotation plan.
35
Keywords: Dusky cotton bug; insecticide; biological traits; cross-resistance; resistance inversion
36 37 38 39 40 41 42 43 44
2
45
1. Introduction
46
Dusky cotton bug Oxycarenus hyalinipennis (Costa) (Hemiptera: Lygaeidae) is a sucking
47
pest of cotton in Pakistan (Jaleel et al., 2014) and commonly referred as the cotton seed bug or
48
cotton stainer bug (Schaefer and Panizzi, 2000). Geographically, O. hyalinipennis is distributed
49
in Asia, Africa, Europe, Central America, the Middle East, South America and the Caribbean
50
(Halbert and Dobbs, 2010). Adult females of O. hyalinipennis lay eggs in cotton lint when bolls
51
open (Kirkpatrick, 1923). Both nymphs and adults of O. hyalinipennis feed on the seeds and suck
52
oil; excessive feeding reduces the seed weight and oil content (Schaefer and Panizzi, 2000). The
53
staining of cotton occurs during picking, and also due to crushing of bugs and their excrement in
54
ginning factories. The fiber becomes dirty due to cast skin and dead insect bodies and may
55
produce an unpleasant smell which ultimately reduces lint quality and market value (Kirkpatrick,
56
1923).
57
Insecticides are the main tools to manage O. hyalinipennis in cotton (Ullah et al., 2015).
58
Insecticides from different chemical groups such as organophosphates, pyrethroids, and
59
neonicotinoids are sprayed in Pakistan multiple times (about 8-10) in the cotton fields during the
60
season (Saeed et al., 2018). As a result of frequent insecticide applications, insecticide resistance
61
has developed in field populations of O. hyalinipennis against various insecticides (Ullah et al.,
62
2015). Studies on insecticide resistance are crucial to know the severity of resistance and to
63
devise suitable pest resistance management strategies. Bifenthrin has been commonly used for
64
both sucking and chewing pests of cotton since 1986 (Ali, 2018). However, the excessive use of
65
this insecticide has resulted in development of resistance in american bollworm Helicoverpa
66
armigera (Hübner) (Lepidoptera: Noctuidae) (Ahmad et al., 1997), spotted bollworm Earias
67
vittella (Fab.) (Lepidoptera: Noctuidae) (Ahmad and Arif, 2009; Jan et al., 2015), cotton
68
mealybug Phenacoccus solenopsis Tinsley (Homoptera: Pseudococcidae) (Saddiq et al., 2014),
69
O. hyalinipennis (Ullah et al., 2015) and cotton jassid Amrasca devastans (Dist.) (Homoptera:
70
Cicadellidae) (Abbas et al., 2018). Moreover, a few studies have reported on bifenthrin
71
resistance in laboratory-selected populations of P. solenopsis (Mansoor et al., 2016) and O.
72
hyalinipennis (Bilal et al., 2018).
73
In resistant genotypes, fitness costs associated with development of resistance are crucial
74
to study as they provide better insight into evolution of insecticide resistance (Gassmann et al.,
75
2009). The biological fitness of resistant population measured in terms of insect survival, 3
76
developmental times, fecundity and net reproductive rates is often reduced following continuous
77
laboratory selection as compared to its counterpart population in the absence of pesticides, hence
78
can be exploited to slow the pace of resistance (Roush and McKenzie, 1987). Fitness costs of
79
pyrethroid resistance have been explored previously against deltamethrin resistance in P.
80
solenopsis (Saddiq et al., 2016a) and Heliothis virescens Fabricius (Lepidoptera: Noctuidae)
81
(Sayyed et al., 2008), and lambda-cyhalothrin resistance in M. domestica (Abbas et al, 2016).
82
Previously, an O. hyalinipennis population was selected with bifenthrin to characterize activities
83
of different detoxificatin enzymes and to evaluate cross-resistance to profenofos, chlorpyrifos,
84
lambda-cyhalothrin, imidacloprid and chlorfenapyr when the selection was stopped (Bilal et al.,
85
2018). However, this study did not provide any information about fitness costs and genetics of
86
bifenthrin resistance. Morover, Bilal et al. (2018) did not study the cross-resistance spectrum to
87
profenofos, deltamethrin and acetamiprid at different selected generations.
88
In this study, we selected field collected O. hyalinipennis with bifenthrin in the laboratory
89
with the aim to verify its resistance development and also to explore cross-resistance possibilities
90
with some other insecticides by conducting bioassays at different generations of O. hyalinipennis
91
during the selection process. The resistant strain was also removed from bifenthrin selection
92
pressure for several generations to determine its impact on resistance stability. We further
93
investigated whether bifenthrin resistance was inherited by sex-linked or autosomal genes and
94
also its extent of dominance. Fitness traits linked with bifenthrin resistance in the resistant and
95
unselected populations of O. hyalinipennis and also in the progeny of their reciprocal crosses
96
were also studied. This study can help with the implementation of insecticide resistance
97
management programs against O. hyalinipennis.
98 99 100
2 Material and methods 2.1. Collection and rearing of insects
101
About 1000 nymphs and adults of O. hyalinipennis were randomly collected from highly
102
infested cotton bolls in a cotton field of the Central Cotton Research Institute (CCRI), Multan
103
(30.19°N, 71.46°E), Pakistan. The field from where the population was collected was sprayed
104
annually about 4-5 times with organophosphate, pyrethroid and neonicotinoids to control O.
105
hyalinipennis (Personal communication with scientific officer). The infested cotton bolls were
106
shaken and insects were collected in plastic jars. After collection, both nymphs and adults were 4
107
separated in the laboratory by using aspirators to obtain a homogenous population to use in
108
bioassays. The population was kept in plastic jars (24 × 12 cm) that were covered with muslin
109
clothes under constant laboratory conditions at 27±2ºC temperature, 60 ± 5% relative humidity
110
and 14:10 L:D hours. Insects were reared on open cotton bolls along with fresh branches of
111
China rose Hibiscus rosasinensis L. as their leaves also provided moisture for the insects. Old
112
branches were replaced with fresh ones twice a week and cotton bolls were changed after every
113
month. The field collected insects were reared for one generation (Go) to eliminate inherent field
114
effects and to increase the population before conducting the bioassays.
115 116
2.2. Populations After acclimatizing the population to laboratory conditions, field collected insects (Field
117
Pop) were divided in two sub-populations; one sub-population was selected with varying
118
concentrations of bifenthrin for 12 generations and designated as “Bifen-Sel” population. The
119
second sub-population was reared parallel to Bifen-Sel population without any insecticide
120
treatment and was designated as “Unsel Pop”. Two reciprocal crosses were also established by
121
crossing the naïve adult insects: 15 males from Bifen-Sel and 15 females from Unsel Pop were
122
crossed to obtain Cross1 progeny and 15 females of Bifen-Sel with 15 males of Unsel Pop were
123
crossed to get Cross2 progeny. Male and female adults were identified by their shape of abdomen
124
i.e. truncate in female and round in male. Moreover, males are shorter in size than females
125
(Henry, 1983).
126
2.3. Insecticides
127
Commercially available formulations of insecticides were used for bioassays: bifenthrin
128
(Talstar® 10EC, FMC, Pakistan), deltamethrin (Decis Super® 10EC, Bayer Crop Sciences,
129
Pakistan), profenofos (Curacron® 500EC, Syngenta, Pakistan) and acetamiprid (Mospilon® 20
130
SP, Arysta Life Sciences, Pakistan).
131
2.4. Bioassays
132
Toxicity bioassays of different insecticides were performed by using the standard IRAC
133
008 leaf-dip method (IRAC, 2019). Two-to-three days old adults of O. hyalinipennis were used
134
in each bioassay. A stock solution (40 mL) was prepared for making five serial concentrations
135
and each concentration was tested in three replicates. Fresh leaves of H. rosasinensis were
136
dipped in each concentration for ten-seconds and air dried for 1-2 hours at room temperature
137
before the insect exposure. Dried leaves were kept in Petri-dishes (5 cm in diameter) on a
5
138
slightly moistened filter paper to prevent the leaves from desiccation. A total of thirty adults
139
(both males and females) were tested for each concentration, ten in each replicate. A total of 180
140
adult insects were used in a bioassay including the control. Insects in the control were allowed to
141
feed on leaves dipped in tap water only. Mortality was assessed after 48 and 72 hours. Adults
142
were considered dead when there was no coordinated movement after a slight touch with a camel
143
hair brush.
144
2.5. Selection protocol for bifenthrin resistance
145
A preliminary bioassay of bifenthrin was performed with the Field Pop to determine the
146
lethal concentrations (LCs) required for selection of O. hyalinipennis with bifenthrin, so that
147
sufficient survivors were left for the next generation. Adults (2-3 days old) were selected with
148
bifenthrin continuously with different lethal concentrations (LC5-LC80) of insecticide ranging
149
from 17-2865 ppm from G1 to G12 (Table 1). The leaf-dip method was used for the selection
150
bioassays by using fresh leaves of H. rosasinensis. For each selection, bifenthrin treated leaves
151
after air drying were kept in Petri-dishes and adults ranging from 160-532 (30-35 per Petri-dish)
152
were exposed. The treated population in Petri-dishes were placed under laboratory conditions at
153
27±2ºC temperature, 60±5% relative humidity and 14:10 h light:dark photoperiod (Ullah et al.,
154
2016). Mortality data were taken after 48 hours exposure to bifenthrin in each selection.
155
Survivors of each selection were reared to get the next progeny for subsequent selection and an
156
average survival of 74.75% was maintained during selections.
157
2.6. Cross-resistance in bifenthrin-selected population
158
The cross-resistance ratio (CRR) in bifenthrin-selected strains was evaluated with other
159
insecticides (mentioned in the insecticides section) as compared to the field population. It was
160
calculated as follows:
161 162
CRR = LC50 of insecticides in Bifen-Sel / LC50 of insecticides in Field Pop 2.7. Stability of bifenthrin resistance
163
The bifenthrin-selected population was reared unselected in the laboratory for four
164
generations from G13-G16 to determine the resistance stability against bifenthrin and some other
165
insecticides. The bioassays with different insecticides were performed at G16 and resistance
166
ratios of Bifen-Sel (G16) were calculated by comparing LC50 of insecticide in Bifen-Sel (G16)
167
with corresponding insecticide LC50 in Unsel Pop.
168
6
169
2.8. Degree of dominance The degree of dominance (DLC) of Bifen-Sel strain was calculated by using the formula
170 171
of Bourguet et al. (2000): = (
1
2 – Unsel Pop) / ( Bifen − Sel − Unsel Pop)
172
Where X is the log of LC50 of tested strain. DLC values vary from 0-1, DLC = 0 indicates
173
completely recessive, DLC = 1 indicates completely dominant nature of insecticide resistance;
174
while DLC 0.50 to <1 and >0<0.50 indicates incompletely dominant and incompletely recessive
175
resistance, respectively.
176
2.9. Procedure to study fitness cost parameters
177
To study the fitness components, a total of 90 newly emerged 3rd instars from Bifen-Sel,
178
Unsel, Cross1 (Bifen-Sel males × Unsel females) and Cross2 (Bifen-Sel females × Unsel males)
179
populations were randomly chosen as starting individuals. Insects from each population were
180
placed in separate plastic jars (14 × 9 cm) containing open cotton bolls and twigs of H.
181
rosasinensis for feeding. The experiment was performed under laboratory conditions as
182
mentioned above. The 90 insects from each population were subdivided into three replicates (30
183
insects each). The following parameters were recorded for each population: developmental
184
duration, survival rate, fecundity and egg hatchability. The fecundity was determined as number
185
of eggs laid by single female until female died and egg hatchability was measured as number of
186
nymphs hatched from eggs. The percent egg hatching was determined as number of nymphs
187
hatched/total eggs ×100 in each replicate. The eggs unable to hatch were considered as non-
188
viable.
189 190
The mean relative growth rate (MRGR) was determined by following formula (Radford, 1967): MRGR = [W2 (mg) – W1 (mg)] / T
191 192
In this equation, W2 and W1 are weights of 1st instar and 5th instar, respectively, and T is time
193
duration from 1st to 5th instar.
194
Net reproductive rate (Ro) was calculated according to Cao and Han (2006): Ro = Nn+1 / Nn
195 196 197 198
Where Nn is the starting generation insects while Nn+1 represents next generation offspring. The Birch (1948) formula was used to calculate intrinsic rate of population increase (rm) as follows: 7
rm = Ro / DT
199 200 201
Where Ro is net reproductive rate and DT is the developmental time from egg to adult. The biotic potential (Bp) was determined using the formula of Roush and Plapp (1982):
202
Bp = [Fecundity (F) / Developmental time ratio (DTr)]
203
Where fecundity (F) was measured as average number of eggs / female and DTr was estimated
204
as DT of tested population / DT of Unsel Pop
205 206 207
The following equation was used to find the relative fitness of resistant and hybrid strains as compared to unselected strain: Relative fitness (Rf) = Ro of Bifen-Sel or Cross1 or Cross2 / Ro of Unsel Pop
208 209
2.10. Data analysis Probit software (version 1.5) was used for analysis of toxicity data (Finney, 1971).
210
Different probit parameters such as LC50, 95% Fiducial limits (FLs), slope with standard errors
211
(SEs) and chi-square (χ2) of each tested insecticide were determined by probit analysis.
212
Statistically, two LC50 values were considered similar if their 95% FLs overlapped (Litchfield
213
and Wilcoxon, 1949). The resistance and cross-resistance ratios determined were categorized as
214
very high (>100), high (51-100), medium (21-50), low (11-20), very low (2-10) and no (<2)
215
resistance (Torres-Vila et al., 2002). The 95% confidence limits (CLs) of resistance and cross-
216
resistance ratios were also determined according to method of Robertson et al. (2007) and
217
considered significant if these did not include the value of 1. The life-history data of all
218
populations were analyzed by statistix software (version 8.1) to estimate relative fitness of tested
219
populations compared to Unsel Pop using completely randomized design. The mean values of
220
different life history data of each population were compared by Least Significant Difference
221
(LSD) test at 5% of significance level.
222 223
3. Results
224
3.1. Toxicities of different insecticides at field and unselected populations and resistance status
225
of field population of O. hyalinipennis
226
For Field Pop, toxicities of profenofos (LC50 = 94.03 ppm) and deltamethrin (LC50 =
227
59.65 ppm) were similar (95% FLs overlapped) but higher (95% FLs did not overlap) than that
228
of bifenthrin (LC50 = 504.2 ppm). Moreover, acetamiprid toxicity (LC50 = 13.6 ppm) was higher
229
(95% FLs did not overlap) than all other insecticides tested for Field Pop. All the insecticides 8
230
had higher toxicities (95% FLs did not overlap) to Unsel Pop than those for Field Pop. The
231
resistance ratios of Field Pop for profenofos, bifenthrin, deltamethrin and acetamiprid were 3.34-
232
, 4.97-, 2.96- and 2.48-fold, respectively, compared to Unsel Pop (Table 2).
233 234 235
3.2. Toxicity and resistance development to bifenthrin in O. hyalinipennis and resistance reversion The LC50s of Bifen-Sel at G3, G5, G7, G9 and G13 using bifenthrin were 890.25, 969.75,
236
1622.94, 4787.03, and 5649.18 ppm, respectively. The Bifen-Sel population of O. hyalinipennis
237
had resistance ratios of 1.77-, 1.92-, 3.21-, 9.49- and 11.20-fold against bifenthrin compared with
238
the Field Pop when tested at the above mentioned generations (Table 2). However, the selected
239
strains showed resistance ratios of 8.77-, 9.55-, 15.98-, 47.14- and 55.64-fold at G3, G5, G7, G9,
240
and G13, respectively, as compared to the Unsel Pop (Table 2). The Bifen-Sel (G13), after rearing
241
for four generations without exposure to bifenthrin, when tested using profenofos, bifenthrin,
242
deltamethrin, and acetamiprid at G16 exhibted resistance ratios of 5.44-, 24.93-, 3.86-, and 4.76-
243
fold, respectively as compared to Unsel Pop. However, toxicities of profenofos, bifenthrin,
244
deltamethrin, and acetamiprid were similar between Bifen-Sel (G16) and Bifen-Sel (G13) for each
245
of these four insecticides (95% FLS overlapped) (Table 2).
246
3.3. Pattern of cross-resistance to Bifen-Sel in O. hyalinipennis
247
The Bifen-Sel population of O. hyalinipennis was evaluated for cross-resistance to
248
profenofos, deltamethrin and acetamiprid at G3, G5, G7, G9, G11 and G13 compared with the Field
249
Pop. The Bifen-Sel population showed no to very low cross-resistance to profenofos (0.77-2.82-
250
fold), deltamethrin (0.85-2.35-fold) and acetamiprid (1.03-2.21-fold) as the selection progressed
251
from G1 to G12 (Table 3).
252
3.4. Resistance and degree of dominance of reciprocal populations
253
The Cross1 and Cross2 populations showed 5.39- and 9.42-fold resistance to bifenthrin,
254
respectively, in comparison to the Unsel Pop. The DLC values of bifenthrin resistance for Cross1
255
and Cross2 were 0.74 and 0.82, respectively indicating the incomplete dominant inheritance of
256
resistance. Moreover, toxicities of bifenthrin were similar (95% FLs overlapped) when tested
257
against both hybrid populations of O. hyalinipennis (Table 4).
258
9
259
3.5. Fitness parameters for Unsel, Bifen-Sel, Cross1 and Cross2 in O. hyalinipennis
260
3.5.1. Developmental durations
261
Mean development durations of different stages in Unsel, Bifen-Sel, Cross1 and Cross2
262
are summarized in Table 5. The developmental duration of eggs did not differ among all tested
263
populations. Developmental duration from 1st to 5th instar in Bifen-Sel was similar to Unsel Pop
264
but significantly higher in Cross1 and Cross2. The adult male longevity of Bifen-Sel, Cross1, and
265
Cross2 was similar to Unsel Pop but it was higher in Cross2 as compared to Cross1. The adult
266
female was significantly higher in Cross2 than that in the other three populations.
267
3.5.2. Survival rates
268
The survival rate of 1st and 2nd instars was significantly lower in Bifen-Sel but similar in
269
Cross1 and Cross2 as compared to Unsel Pop (Table 5). The survival rate of 3rd instar was
270
significantly lower in Bifen-Sel and Cross1 than that of Unsel Pop. No significant differences
271
were found in the survival of 4th and 5th instars in all tested populations. The average survival rate
272
from 1st to 5th instar was significantly lower in Bifen-Sel compared to the other three
273
populations.
274
3.5.3. Fecundity, egg hatchability, net reproductive rate (Ro), intrinsic rate of population
275
increase (rm) and relative fitness (Rf)
276
The Bifen-Sel females produced significantly lower number of eggs compared with the
277
Unsel Pop, Cross1 and Cross2. The Unsel pop had a significantly lower fecundity than Cross2 but
278
not than Cross1. The percent egg hatchability and net reproductive rate (Ro) in the Bifen-Sel
279
were significantly less when compared with the other three populations. The intrinsic rate of
280
population increase (rm) was significantly lower in Bifen-Sel compared to all other populations
281
which had similar rm. The relative fitness (Rf) of Bifen-Sel was significantly lower than that of
282
the other three populations. Cross1 had a similar Rf than Unsel Pop. Cross2 had a significantly
283
higher Rf than Unsel Pop (Table 6).
284
3.5.4. Biotic potential (Bp) and mean relative growth Rate (MRGR)
285
The Bp of Bifen-Sel was significantly lower in comparison with the Unsel Pop. However
286
hybrid populations showed similar Bp to Unsel Pop (F = 15.7; P = 0.0010; df = 3,8) (Fig 1). A
287
significantly higher MRGR was found in all the tested populations as compared to Unsel Pop (F
288
= 52.8; P<0.0001; df = 3,8) (Fig 1).
289
10
290
4. Discussion
291
In the current study, selection of O. hyalinipennis for 12 generations resulted in a
292
substantial increase in LC50 and resistance ratios to bifenthrin. The LC50 of bifenthrin in the Field
293
Pop at G1 was 504.2 ppm with very low resistance (4.97-fold) before selection in the laboratory.
294
However, when the same population was continuously selected until G12, the LC50 of bifenthrin
295
in Bifen-Sel population (G13) increased to 5649.18 ppm or 55.64-fold compared with the Unsel
296
Pop. Laboratory induced high to very high levels of resistance selections have been reported in
297
the house fly Musca domestica Linnaeus (Diptera: Muscidae) to beta-cypermethrin with 4420-
298
fold resistance after 25 selected generations compared with the susceptible strain (Zhang et al.,
299
2008), 474-fold resistance to cypermethrin in H. armigera over six selected generations
300
compared with the laboratory population (Achaleke and Brévault, 2010), 236-fold resistance to
301
deltamethrin in Plutella xylostella (L.) (Lepidoptera: Plutellidae) after six selected generations
302
compared with the unselected population (Sayyed et al., 2005), and 54.32-fold and 178.42-fold
303
resistance to bifenthrin in P. solenopsis after 14 generations when compared with the field and
304
laboratory strains, respectively (Mansoor et al., 2016). The resistance levels selected under
305
laboratory conditions can be low or high depending upon the selection pressure, previous history
306
of exposure to insecticides, insect species, selection protocols, geographic origins of populations
307
and presence of susceptible and resistant genes.
308
In this study, the decline in bifenthrin resistance from 55.64-fold to 24.93-fold (about
309
55% decrease) after a few non-selected generations with the Bifen-Sel population suggested the
310
unstable nature of bifenthrin resistance in O. hyalinipennis. Similar to our results, reversion of
311
insecticide resistance has been reported previously in O. hyalinipennis after five unexposed
312
generations against chlorfenapyr (Ullah and Shad, 2017) and bifenthrin (Bilal et al., 2018).
313
Recovery of resistant populations towards susceptibility could be due to high fitness costs
314
associated with insecticide resistance (Gassmann et al., 2009) which are in agreement with the
315
present findings.
316
.The Bifen-Sel population of O. hyalinipennis did not show any cross-resistance to
317
profenofos and deltamethrin up to the fifth selected generation. However, as the selection
318
continued to the 12th generation, there was a slight increase (very low) in cross-resistance against
319
both profenofos (2.01-2.82-fold), and deltamethrin (2.00-2.35-fold) when bioassayed at G13.
320
Similarly, the Bifen-Sel population did not have any cross-resistance to acetamiprid until at G7 11
321
but a very low cross-resistance (2.35-, 2.11-, and 2.21-fold) was expressed at G9, G11, and G13.
322
Lack of and/or very low cross-resistance in the Bifen-Sel population against profenofos and
323
acetamiprid was expected as both of these insecticides have entirely different mode of actions
324
and belong to different chemical groups than bifenthrin. Profenofos is an organophosphorus
325
insecticide and is known to act as acetylcholinesterase (AChE) inhibitor at synapse while
326
acetamiprid is neonicotinoid group insecticide and works as nicotinic acetylcholine receptor
327
(nAChR) competitive modulator. However, bifenthrin is a pyrethroid insecticide and targets the
328
axon of neuron as sodium channel modulator (IRAC, 2019). Unexpectedly, presence of very low
329
cross-resistance in the Bifen-Sel population of O. hyalinipennis against another pyrethroid
330
(deltamethrin) might be due to limited exposure of the pest population to deltamethrin in the
331
field and involvement of several resistance mechanisms. A deltamethrin-selected population
332
(4632.8-fold) of P. solenopsis had no cross-resistance to profenofos but a low level of cross-
333
resistance against lambda-cyhalothrin and acetamiprid (Afzal et al., 2018). Results of this study
334
are similar to Mansoor et al. (2016) who found negligible cross-resistance in a bifenthrin selected
335
population of P. solenopsis against buprofezin, chlorpyrifos and lambda-cyhalothrin.
336
Insect populations often experience a decline in biological fitness during the development
337
of resistance. These phenomena have been found in many insecticide resistance studies such as
338
in P. xylostella to Bacillus thuringiensis toxin Cry2Ad (Liao et al., 2019), brown planthopper
339
Nilaparvata lugens (Stål) (Hemiptera: Delphacidae) to nitenpyram (Zhang et al., 2018), M.
340
domestica to spinosad (Khan, 2018), Spodoptera litura (Fab.) (Lepidoptera: Noctuidae) to
341
bistrifluron (Huang et al., 2019), fall armyworm Spodoptera frugiperda (J.E. Smith)
342
(Lepidoptera: Noctuidae) to Bt toxin Cry1A.105 (Niu et al., 2018), green peach aphid Myzus
343
persicae (Sulzer) (Hemiptera: Aphididae) to sulfoxaflor (Wang et al., 2018), H. armigera to
344
indoxacarb (Cui et al., 2018), western flower thrips Frankliniella occidentalis (Thysanoptera:
345
Thripidae) and maize armyworm Mythimna separata (Lepidoptera: Noctuidae) to thiamethoxam
346
(Gao et al., 2014; Yasoob et al., 2018), and P. solenopsis to spirotetramat (Ejaz and Shad, 2017).
347
In this study, bifenthrin resistance development in the Bifen-Sel population resulted in declines
348
in nymphal survival, fecundity, egg hatchability, Ro, rm, and Rf (0.58) as compared with Unsel
349
Pop. These findings indicate that, if selection discontinued, the Bifen-Sel population might not
350
increase as rapidly as the Unsel Pop because of significant disadvantages in life history
351
parameters. These results also suggest the presence of a trade-off in distribution of energy 12
352
resources between costs in the form of fitness and mechanism(s) of resistance. Similar to these
353
results, lower relative fitness due to insecticide resistance has been reported in other insects
354
under different selection regimes such as in yellow fever mosquito Aedes aegypti (Linnaeus)
355
(Diptera: Culicidae) (Jaramillo-O et al., 2014) and M. domestica (Abbas et al., 2016) to lambda-
356
cyhalothrin resistance; and P. solenopsis to deltamethrin resistance (Saddiq et al., 2016a).
357
Determination of fitness costs associated with resistance is important to study in
358
homozygous resistant individuals but also in heterozygotes that can act as a carrier of most
359
abundant resistant genes in early stages of resistance (Jia et al., 2009). Owing to this, fitness
360
costs of bifenthrin resistance were also estimated in two hybrid populations (Cross1 and Cross2)
361
of Bifen-Sel and Unsel Pop. The results showed that the Rf of Cross1 (1.07) and Cross2 (1.22)
362
were similar and both population progenies had superior fitness traits when compared with the
363
the parental Bifen-Sel. Moreover, in both hybrids, fitness recovery was significantly higher in
364
Cross2 compared with the Unsel Pop. The advantageous biological characteristics such as
365
survival, fecundity, egg hatchability, Ro and rm in reciprocal cross progenies compared to Bifen-
366
Sel population might be attributed to their increased vigor, or heterosis, which might have
367
resulted in longer inbreeding leading to the improvement of beneficial traits in heterozygous
368
progeny. Therefore, the study of life history parameters of both resistant and reciprocal
369
individuals is crucial and helpful in formulating resistance management strategies (Jia et al.,
370
2009).
371
The hybrid progenies with resistance ratios of 5.39-fold (Cross1) and 9.42-fold (Cross2)
372
were also used to determine inheritance characteristics associated with bifenthrin resistance. The
373
overlapping of FLs of LC50 values in both populations suggested that maternal effects were not
374
involved in bifenthrin resistance in O. hyalinipennis and it was autosomal. Moreover, DLC of
375
both populations was >0.50 and <1, which indicated that bifenthrin resistance was incompletely
376
dominant. Autosomal and incompletely dominant resistance have also been reported in many
377
insect pests (Sayyed et al., 2005; Achaleke and Brévault, 2010; Khan et al., 2015; Saddiq et al.
378
2016b; Ullah et al., 2016; Ijaz and Shad, 2018).
379
5. Conclusion and recommendations
380
The present findings showed that O. hyalinipennis has a high potential to develop
381
bifenthrin resistance following continuous selection pressure. The high resistance was also
382
reverted upon removal of selection pressure and no obvious cross-resistance occured with any 13
383
tested insecticide. Moreover, fitness costs were apparent in the resistant population. Bifenthrin
384
resistance inheritance appeared to be autosomal and incompletely dominant. The unstable
385
bifenthrin resistance indicated that evolution of resistance to bifenthrin may be minimized in O.
386
hyalinipennis if the insecticide is removed from a management program for a given duration. A
387
very weak cross-resistance, high fitness costs and incomplete dominance of bifenthrin resistance
388
suggests that these insecticides can be included in a rotation program to delay resistance
389
development. Recovery of fitness traits in hybrid populations also indicated that mixing the
390
selected (resistant) and unselected populations might suppress the resistance by diluting the
391
resistant genes.
392 393
14
394 395 396
Conflict of interest statement No potential conflict of interest exists among all authors Author contributions
397
AB conceived the study. MBSA and SAS designed the study. AB collected the insects
398
and carried out the research work. SAS supervised the laboratory work. MBSA provided
399
technical guidelines to process the raw data. AB analyzed the data. AB and MBSA wrote the
400
manuscript. SAS read and approved the written manuscript.
401 402
Acknowledgements We acknowledge Prof. Dr. José Eduardo Serrão, Department of General
403
Federal University of Viçosa, Brazil, and Prof. (Retd.) Dr. Muhammad Aslam, Department of
404
Entomology, Faculty of Agricultural Sciences and Technology, Bahauddin Zakariya University,
405
Multan, Pakistan for sparing their time to check manuscript for improvement of English
406
language and sense.
407
15
Biology,
408
References
409
Abbas, N., Ismail, M., Ejaz, M., Asghar, I., Irum, A., Shad, S.A., Binyameen, M., 2018.
410
Assessment of field evolved resistance to some broad-spectrum insecticides in cotton
411
jassid, Amrasca devastans from southern Punjab, Pakistan. Phytoparasitica 46, 411–419.
412
Abbas, N., Shah, R.M., Shad, S.A., Iqbal, N., Razaq, M., 2016. Biological trait analysis and
413
stability of lambda-cyhalothrin resistance in the house fly, Musca domestica L. (Diptera:
414
Muscidae). Parasitol. Res. 115, 2073–2080.
415
Achaleke, J., Brévault, T., 2010. Inheritance and stability of pyrethroid resistance in the cotton
416
bollworm Helicoverpa armigera (Lepidoptera: Noctuidae) in Central Africa. Pest
417
Manage. Sci. 66, 137–141.
418 419
Afzal MBS, Shad SA, 2015. Resistance inheritance and mechanism to emamectin benzoate in Phenacoccus solenopsis (Homoptera: Pseudococcidae). Crop Prot. 71, 60–65
420
Afzal, M.B.S., Shad, S.A., Ejaz, M., Ijaz, M., 2018. Selection, cross-resistance, and resistance
421
risk assessment to deltamethrin in laboratory selected Phenacoccus solenopsis
422
(Homoptera: Pseudococcidae). Crop Prot. 112, 67–73.
423
Ahmad, M., Arif, M.I., 2009. Resistance of Pakistani field populations of spotted bollworm
424
Earias vittella (Lepidoptera: Noctuidae) to pyrethroid, organophosphorus and new
425
chemical insecticides. Pest Manage. Sci. 65, 433–439.
426 427
Ahmad, M., Arif, M.I., Attique, M.R., 1997. Pyrethroid resistance of Helicoverpa armigera (Lepidoptera: Noctuidae) in Pakistan. Bull. Entomol. Res. 87, 343–347.
428
Ali, M.A., 2018. The pesticides registered with recommendations for safe handling and use in
429
pakistan, a hand book for agriculture extension agents. Published by Directorate General
430
Agriculture (Extension & AR) Punjab, Lahore.
431
Bilal, M., Freed, S., Ashraf, M.Z., Rehan, A., 2018. Resistance and detoxification enzyme
432
activities to bifenthrin in Oxycarenus hyalinipennis (Hemiptera: Lygaeidae). Crop Prot.
433
111, 17–22.
434 435 436 437
Birch, L., 1948. The intrinsic rate of natural increase of an insect population. J. Anim. Ecol. 17, 15–26. Bourguet, D., Genissel, A., Raymond, M., 2000. Insecticide resistance and dominance levels. J. Econ. Entomol. 93, 1588–1595.
16
438 439 440
Cao, G., Han, Z., 2006. Tebufenozide resistance selected in Plutella xylostella and its cross‐resistance and fitness cost. Pest Manage. Sci. 62, 746–751. Cui, L., Wang, Q., Qi, H., Wang, Q., Yuan, H., Rui, C., 2018. Resistance selection of indoxacarb
441
in
Helicoverpa
armigera
(Hübner)
(Lepidoptera:
Noctuidae):
cross‐resistance,
442
biochemical mechanisms and associated fitness costs. Pest Manage. Sci. 74, 2636–2644.
443
Ejaz, M., Shad, S.A, 2017. Spirotetramat resistance selected in the Phenacoccus solenopsis
444
(Homoptera: Pseudococcidae): cross-resistance patterns, stability, and fitness costs
445
analysis. J. Econ. Entomol. 110, 1226–1234.
446
Finney, D.J., 1971. Probit Analysis, third ed. Cambridge University Press, UK, 333 pp.
447
Gao, C.F., Ma, S.Z., Shan, C.H., Wu, S.F., 2014. Thiamethoxam resistance selected in the
448
western flower thrips Frankliniella occidentalis (Thysanoptera: Thripidae): Cross-
449
resistance patterns, possible biochemical mechanisms and fitness costs analysis. Pestic.
450
Biochem. Physiol. 114, 90–96.
451 452
Gassmann, A.J., Carrière, Y., Tabashnik, B.E., 2009. Fitness costs of insect resistance to Bacillus thuringiensis. Ann. Rev. Entomol. 54, 147–163.
453
Halbert, S.E., Dobbs, T., 2010. Cotton Seed Bug, Oxycarenus hyalinipennis (Costa): A serious
454
pest of cotton that has become established in the Caribbean Basin. FDACS-Pest Alert
455
DACS-P-01726, Florida Department of Agriculture and Consumer Services, Division of
456
Plant Industry [Online] http://www. freshfromflorida. com/pi/pest_alerts/pdf/cotton-
457
seedbug-pest-alert. pdf. Accessed 6, 2012.
458 459
Henry, T.J., 1983. Pests not known to occur in the United States or of limited distribution, USDA-APHIS-PPQ, APHIS 43, 1–6.
460
Huang, Q., Wang, X., Yao, X., Gong, C., Shen, L., 2019. Effects of bistrifluron resistance on the
461
biological traits of Spodoptera litura (Fab.) (Noctuidae: Lepidoptera). Ecotoxicol. 28,
462
323–332.
463
Ijaz, M., Shad, S.A., 2018. Inheritance mode and realized heritability of resistance to
464
imidacloprid in Oxycarenus hyalinipennis Costa (Hemiptera: Lygaeidae). Crop Prot. 112,
465
90–95.
466
IRAC, 2019. IRAC Mode of Action Classification Scheme.
17
467
Jaleel, W., Saeed, S., Naqqash, M.N., Zaka, S.M., 2014. Survey of Bt cotton in Punjab Pakistan
468
related to the knowledge, perception and practices of farmers regarding insect pests. Int.
469
J. Agric. Crop Sci. 7, 10–20.
470
Jan, M.T., Abbas, N., Shad, S.A., Saleem, M.A., 2015. Resistance to organophosphate,
471
pyrethroid and biorational insecticides in populations of spotted bollworm, Earias vittella
472
(Fabricius) (Lepidoptera: Noctuidae), in Pakistan. Crop Prot. 78, 247–252.
473
Jaramillo-O. N., Fonseca-Gonzalez, I., Chaverra-Rodrıguez, D., 2014. Geometric morphometrics
474
of nine field isolates of Aedes aegypti with different resistance levels to lambda-
475
cyhalothrin and relative fitness of one artificially selected for resistance. PLoS ONE 9,
476
96379.
477
Jia, B., Liu, Y., Zhu, Y.C., Liu, X., Gao, C., Shen, J., 2009. Inheritance, fitness cost and
478
mechanism of resistance to tebufenozide in Spodoptera exigua (Hübner) (Lepidoptera:
479
Noctuidae). Pest Manag. Sci. 65, 996–1002.
480
Khan, H. A.A., Akram, W., Haider, M.S. 2015. Genetics and mechanism of resistance to
481
deltamethrin in the house fly, Musca domestica L., from Pakistan. Ecotoxicol. 24, 1213–
482
1220.
483 484 485 486
Khan, H.A.A., 2018. Spinosad resistance affects biological parameters of Musca domestica Linnaeus. Sci. Rep. 8, 14031. Kirkpatrick, T., 1923. The Egyptian Cotton Seed Bug. Ministry Agricult. Egypt., Techn. a. Sci. Serv. Bull. 35, 1–106.
487
Liao, J., Xue, Y., Xiao, G., Xie, M., Huang, S., You, S., Wyckhuys, K.A., You, M., 2019.
488
Inheritance and fitness costs of resistance to Bacillus thuringiensis toxin Cry2Ad in
489
laboratory strains of the diamondback moth, Plutella xylostella (L.). Sci. Rep. 9, 6113.
490 491
Litchfield, J.T., Wilcoxon, F., 1949. A simplified method of evaluating dose–effect experiments. J. Pharmacol. Exp. Theory 99, 99–103.
492
Mansoor, M.M., Afzal, M.B.S., Basoalto, E., Raza, A.B.M., Banazeer, A., 2016. Selection of
493
bifenthrin resistance in cotton mealybug Phenacoccus solenopsis Tinsley (Homoptera:
494
Pseudococcidae):
495
mechanism. Crop Prot. 87, 55–59.
496 497
Cross-resistance,
realized
heritability and
possible
resistance
Niu, Y., Head, G.P., Price, P.A., Huang, F., 2018. Inheritance and fitness costs of Cry1A. 105 resistance in two strains of Spodoptera frugiperda (JE Smith). Crop Prot. 110, 229–235. 18
498
Radford, P.J., 1967. Growth Analysis Formulae-Their Use and Abuse 1. Crop Sci. 7, 171–175.
499
Robertson, J.L., Savin, N., Preisler, H.K., Russell, R.M., 2007. Bioassays with Arthropods,
500 501 502 503 504
second ed. CRC press, Boca Raton, Florida. Roush, R.T., McKenzie, J.A., 1987. Ecological genetics of insecticide and acaricide resistance. Ann. Rev. Entomol. 32, 361–380. Roush, R.T., Plapp, F.W., 1982. Effects of insecticide resistance on biotic potential of the house fly (Diptera: Muscidae). J. Econ. Entomol. 75, 708–713.
505
Saddiq, B., Abbas, N., Shad, S.A., Aslam, M., Afzal, M.B.S., 2016a. Deltamethrin resistance in
506
the cotton mealybug, Phenacoccus solenopsis Tinsley: cross-resistance to other
507
insecticides, fitness cost analysis and realized heritability. Phytoparasitica 44, 83–90.
508
Saddiq, B., Afzal, M.B.S., Shad, S.A. 2016b. Studies on genetics, stability and possible
509
mechanism of deltamethrin resistance in Phenacoccus solenopsis Tinsley (Homoptera:
510
Pseudococcidae) from Pakistan. J.Genet. 95, 1009–1016.
511
Saddiq, B., Shad, S.A., Khan, H.A.A., Aslam, M., Ejaz, M., Afzal, M.B.S., 2014. Resistance in
512
the mealybug Phenacoccus solenopsis Tinsley (Homoptera: Pseudococcidae) in Pakistan
513
to selected organophosphate and pyrethroid insecticides. Crop Prot. 66, 29–33.
514
Saeed, R., Abbas, N., Razaq, M., Mahmood, Z., Naveed, M., Rehman, H.M.U., 2018. Field
515
evolved resistance to pyrethroids, neonicotinoids and biopesticides in Dysdercus koenigii
516
(Hemiptera: Pyrrhocoridae) from Punjab, Pakistan. Chemosphere 213, 149–155.
517
Sayyed, A.H., Ahmad, M., Crickmore, N., 2008. Fitness costs limit the development of
518
resistance to indoxacarb and deltamethrin in Heliothis virescens (Lepidoptera:
519
Noctuidae). J. Econ. Entomol. 101, 1927–1933.
520
Sayyed, A.H., Attique, M.N.R., Khaliq, A., Wright, D.J., 2005. Inheritance of resistance and
521
cross‐resistance to deltamethrin in Plutella xylostella (Lepidoptera: Plutellidae) from
522
Pakistan. Pest Manage. Sci. 61, 636–642.
523 524 525 526
Schaefer, C.W., Panizzi, A.R., 2000. Economic importance of Heteroptera: a general view, Heteroptera of economic importance. CRC Press, pp. 25-30. Schaefer, C.W., Panizzi, A.R., 2000. Economic importance of Heteroptera: a general view, Heteroptera of economic importance. CRC Press, pp. 25–30.
527 528
19
529
Torres-Vila, L. M., Rodrı́guez-Molina, M. C., Lacasa-Plasencia, A., Bielza-Lino, P., Rodrı́guez-
530
del-Rincón, Á., 2002. Pyrethroid resistance of Helicoverpa armigera in Spain: current
531
status and agroecological perspective. Agric. Ecosys. Environ. 93, 55–66.
532
Ullah, S., Shad, S.A., Abbas, N., 2015. Resistance of dusky cotton bug, Oxycarenus
533
hyalinipennis Costa (Lygaidae: Hemiptera), to conventional and novel chemistry
534
insecticides. J. Econ. Entomol. 109, 345–351.
535
Ullah, S., Shah, R.M., Shad, S.A., 2016. Genetics, realized heritability and possible mechanism
536
of chlorfenapyr resistance in Oxycarenus hyalinipennis (Lygaeidae: Hemiptera). Pestic.
537
Biochem. Physiol. 133, 91–96.
538
Ullah, S., Shad, S.A., 2017. Toxicity of insecticides, cross-resistance and stability of
539
chlorfenapyr resistance in different strains of Oxycarenus hyalinipennis Costa
540
(Hemiptera: Lygaeidae). Crop Prot. 99, 132–136.
541
Wang, Z.H., Gong, Y.J., Chen, J.C., Su, X.C., Cao, L.J., Hoffmann, A.A., Wei, S.J., 2018.
542
Laboratory selection for resistance to sulfoxaflor and fitness costs in the green peach
543
aphid Myzus persicae. J. Asia-Pacific Entomol. 21, 408–412.
544
Yasoob, H., Abbas, N., Li, Y., Zhang, Y., 2018. Selection for resistance, life history traits and
545
the biochemical mechanism of resistance to thiamethoxam in the maize armyworm,
546
Mythimna separata (Lepidoptera: Noctuidae). Phytoparasitica 46, 627–634.
547 548
Zhang, L., Shi, J., Gao, X., 2008. Inheritance of beta‐cypermethrin resistance in the housefly Musca domestica (Diptera: Muscidae). Pest Manage. Sci. 64, 185–190.
549
Zhang, X., Mao, K., Liao, X., He, B., Jin, R., Tang, T., Wan, H., Li, J., 2018. Fitness cost of
550
nitenpyram resistance in the brown planthopper Nilaparvata lugens. J. Pest Sci. 91,
551
1145–1151.
20
552
Table 1. History of selection of Oxycarenus hyalinipennis with bifenthrin in the laboratory Generation G1 G2 G3 G4 G5 G6 G7 G8 G9 G10 G11 G12
Concentration (ppm) 17 35 59 172 504.21 504.21 653.55 850.67 1,000 1324.2 2029.4 2865
No exposed 400 370 524 532 407 200 160 220 180 300 281 411
No died 45 46 27 17 273 60 64 39 30 40 91 221
No survived 355 324 497 515 134 140 96 181 150 260 190 190
553
21
%Mortality 11.25 12.43 5.43 3.20 67.08 30.00 40.00 17.73 16.67 13.33 32.38 53.77
%Survival 88.75 87.57 94.85 96.80 32.92 70.00 60.00 82.27 83.33 86.67 67.62 46.23
Table 2. Response of different populations of Oxycarenus hyalinipennis to tested insecticides
a
Population
Insecticide
LC50 (ppm)
95% FL (ppm)
Slope(±S.E)
χ2
DF
P
Na
RRb(95%CL)
RRc(95%CL)
Field Pop (G1)
Profenofos
94.03
70.88-125.91
1.89(±0.29)
0.24
4
0.99
180
1
3.34(2.24-4.98)
Field Pop (G1)
Bifenthrin
504.2
355.018-852.924
1.12(±0.20)
1.2
4
0.88
180
1
4.97(3.00-8.21)
Field Pop (G1)
Deltamethrin
59.65
41.18-98.71
1.34(±0.27)
2.79
4
0.59
180
1
2.96(1.76-4.98)
Field Pop (G1)
Acetamiprid
13.6
9.00-28.84
1.48(±0.30)
0.67
4
0.95
180
1
2.48(1.38-4.48)
Unsel Pop
Profenofos
28.14
21.11-38.07
1.84(±0.29)
1.85
4
0.76
180
1
1
Unsel Pop
Bifenthrin
101.54
75.83-138.77
1.79(±0.29)
1.5
4
0.83
180
1
1
Unsel Pop
Deltamethrin
20.17
14.86-29.69
1.68(±0.28)
2.32
4
0.68
180
1
1
Unsel Pop
Acetamiprid
5.48
4.13-7.35
1.88(±0.29)
1.55
4
0.82
180
1
1
Bifen-Sel (G3)
Bifenthrin
890.25
626.93-1394.28
1.42(±0.27)
1
4
0.91
180
1.77(1.11-3.18)
8.77(4.65-12.26)
Bifen-Sel (G5)
Bifenthrin
969.75
710.99-1430.41
1.64(±0.29)
1.15
4
0.89
180
1.92(1.13-3.26)
9.55(6.13-14.89)
Bifen-Sel (G7)
Bifenthrin
1622.94
1143.40-2698.50
1.46(±0.28)
0.72
4
0.95
180
3.21(1.82-5.70)
15.98(9.76-26.18)
Bifen-Sel (G9)
Bifenthrin
4787.03
3062.35-11650.08
1.44(±0.31)
0.31
4
0.99
180
9.49
47.14(24.57-90.46)
Bifen-Sel (G13)
Bifenthrin
5649.18
3680.06-12389.88
1.27(±0.28)
0.67
4
0.95
180
11.20(5.72-21.95)
55.64(30.29-102.19)
Bifen-Sel (G13)
Profenofos
265.1
187.33-450.10
1.53(±0.29)
1.17
4
0.88
180
2.82(1.73-4.61)
9.42(5.75-15.45)
Bifen-Sel (G13)
Deltamethrin
140.01
102.53-221.17
1.78(±0.31)
0.57
4
0.97
180
2.35(1.37-4.03)
6.94(4.27-11.30)
Bifen-Sel (G13)
Acetamiprid
30.04
21.84-47.13
1.65(±0.30)
1.94
4
0.75
180
2.21(1.17-4.16)
5.48(3.48-8.66)
Bifen-Sel (G16)
Profenofos
153.02
106.91-275.02
1.58(±0.30)
0.75
4
0.83
180
-
-
Bifen-Sel (G16)
Bifenthrin
2531.55
1663.38-5578.06
1.45(±0.30)
0.33
4
0.99
180
-
-
Bifen-Sel (G16)
Deltamethrin
77.91
54.18-139.41
1.63(±0.30)
1.29
4
0.86
180
-
-
Bifen-Sel (G16)
Acetamiprid
26.1
18.47-45.37
1.49(±0.29)
1.26
4
0.87
180
-
-
Number of adult insects treated in each bioassay including control Resistance ratio = LC50 of insecticides in selected population / LC50 of insecticides in Field Pop c Resistance ratio = LC50 of insecticides in Field or Bifen-Sel population (G3 to G13) / LC50 of insecticides in Unsel Pop d Resistance ratio = LC50 of insecticide in Bifen-Sel population (G16) / LC50 of insecticide in Unsel Pop b
22
RRd(95%CL)
5.44(3.24-9.12) 24.93(13.55-45.90) 3.86(2.25-6.64) 4.76(2.90-7.85)
Table 3. Cross-resistance evaluation against some insecticides at different generations of Bifen-Sel population of Oxycarenus hyalinipennis Population Insecticide LC50 (ppm) 95% FL (ppm) Slope (±S.E) χ2 DF P Na CRRb(95%CL) Field Pop (G1) Profenofos 94.03 70.88-125.91 1.89(±0.29) 0.24 4 0.99 180 Field Pop (G1) Deltamethrin 59.65 41.18-98.71 1.34(±0.27) 2.79 4 0.59 180 Field Pop (G1) Acetamiprid 13.6 9.00-28.84 1.48(±0.30) 0.67 4 0.95 180 74.25 54.02-99.065 1.81(±0.29) 3.40 4 0.49 180 0.79(0.53-1.18) Bifen-Sel (G3) Profenofos Bifen-Sel (G5) Profenofos 72.85 50.37-100.69 1.57(±0.28) 1.10 4 0.89 180 0.77(0.50-1.20) Bifen-Sel (G7) Profenofos 189.19 139.91-283.94 1.74(±0.30) 0.24 4 0.99 180 2.01(1.30-3.11) Bifen-Sel (G9) Profenofos 195.74 147.20- 275.24 1.85(±0.30) 0.45 4 0.98 180 2.08(1.38-3.14) Bifen-Sel (G11) Profenofos 244.21 175.44-393.77 1.51(±0.29) 1.15 4 0.89 180 2.60(1.63-4.15) Bifen-Sel (G13) Profenofos 265.1 187.33-450.10 1.53(±0.29) 1.17 4 0.88 180 2.82(1.73-4.61) Bifen-Sel (G3) Deltamethrin 50.73 37.79-71.26 1.75(±0.29) 0.61 4 0.96 180 0.85(0.51-1.41) Bifen-Sel (G5) Deltamethrin 58.37 43.32-83.45 1.72(±0.29) 0.69 4 0.95 180 0.98(0.59-1.63) Bifen-Sel (G7) Deltamethrin 142.90 86.38- 422.80 1.12(±0.27) 0.4 4 0.98 180 2.40(1.10-5.21) Bifen-Sel (G9) Deltamethrin 152.84 94.76-406.57 1.18(±0.28 0.92 4 0.92 180 2.56(1.22-5.37) Bifen-Sel (G11) Deltamethrin 119.34 85.07-198.25 1.57(±0.29) 0.04 4 1.00 180 2.00(1.14-3.52) Bifen-Sel (G13) Deltamethrin 140.01 102.53-221.17 1.78(±0.31) 0.57 4 0.97 180 2.35(1.37-4.03) Bifen-Sel (G3) Acetamiprid 14.74 9.86-30.68 1.51(±0.31) 0.59 4 0.96 180 1.08(0.52-2.24) Bifen-Sel (G5) Acetamiprid 14.04 9.52-28 1.54(±0.31) 0.63 4 0.96 180 1.03(0.51-2.10) Bifen-Sel (G7) Acetamiprid 17 12.63-25.39 1.79(±0.30) 1.27 4 0.87 180 1.25(0.68-2.32) Bifen-Sel (G9) Acetamiprid 31.95 21.07-68.05 1.35(±0.29) 0.13 4 1.00 180 2.35(1.13-4.90) Bifen-Sel (G11) Acetamiprid 28.67 20.69-45.00 1.62(±0.29) 1.45 4 0.84 180 2.11(1.11-4.00) Bifen-Sel (G13) Acetamiprid 30.04 21.84-47.13 1.65(±0.30) 1.94 4 0.75 180 2.21(1.17-4.16) a Number of adult insects treated in each bioassay including control b Cross-resistance ratio = LC50 of each insecticide in Bifen-Sel / LC50 of corresponding insecticide in Field Pop
23
Table 4. Bifenthrin resistance and its dominance in reciprocal crosses DF P Na RRb(95% CL) DLCc Population LC50 (ppm) 95 % FL (ppm) Slope±SE χ2 Unsel Pop 101.54 75.83-138.77 1.79±0.29 1.50 4 0.83 180 1 Cross1 547.14 404.45-750.09 1.74±0.29 1.04 4 0.90 180 5.39(3.55-8.18) 0.74 956.10 704.39-1448.1 1.72±0.30 1.19 4 0.88 180 9.42(6.02-14.74) 0.82 Cross2 a Number of adult insects treated in each bioassay including control b Resistance ratio = LC50 of bifenthrin in Cross1 or Cross2 / LC50 of bifenthrin in Unsel Pop c Degree of dominance
24
Table 5. Developmental durations in days and survival rates (%) of different stages in different tested populations of Oxycarenus hyalinipennis Mean ±SE Unsel Pop Bifen-Sel Cross1 Cross2 Egg duration 5.16 ±0.47 A 5.06 ±0.89 A 5.27 ±0.18 A 5.51 ±0.37 A st th 1 to 5 instar duration 18.52 ±0.25 B 19.67 ±0.56 AB 21.15 ±0.31 A 20.05 ±0.63 A Egg to adult duration 23.68 ±0.32B 24.73 ±0.48 AB 26.420 ±0.42 A 25.56 ±1.00 AB Adult male longevity 4.8 ±0.29 AB 5.92 ±0.36 AB 4.44 ±0.80 B 6.11 ±0.11 A Adult female longevity 11.31 ±0.72 B 11.22 ±0.97 B 13.33 ±1.20 B 17.36 ±0.81 A 1st instar survival 82.03 ±2.34 A 73.28 ±1.46 B 84.85 ±1.32 A 84.75 ±1.84 A nd 2 instar survival 85.95 ±2.64 A 76.20 ±2.73 B 83.10 ±1.58 AB 80.99 ±1.76 AB rd 3 instar survival 97.78 ±1.11 A 91.11 ±2.22 B 90 ±1.92 B 94.44 ±1.11 AB th 4 instar survival 98.89 ±1.11 A 96.34 ±2.06 A 96.29 ±0.08 A 98.85 ±1.15 A th 5 instar survival 100 ±0.00 A 97.62 ±2.38 A 98.72 ±1.28 A 96.43 ±0.00 A Av. instar survival 92.87 ±0.18 A 86.91 ±0.75 B 90.59 ±1.01 A 91.09 ±0.58 A The means within a row denoted by similar letters are not significantly different (P>0.05). df values for ANOVA = 3,8 Biological traits
25
ANOVA parameters F = 0.13; P = 0.9416 F = 5.47; P = 0.0243 F = 3.63; P = 0.0645 F = 3.08; P = 0.0901 F = 9.27; P = 0.0055 F = 9.34; P = 0.0054 F = 3.38; P = 0.0746 F = 4.44; P = 0.0407 F = 1.28; P = 0.3466 F = 1.27; P = 0.3473 F = 9.68; P = 0.0049
Table 6. Comparison of means ±SE of fecundity, hatchability, Ro, rm, and and Rf of Unsel, Bifen-Sel, Cross1 and Cross2 populations of Oxycarenus hyalinipennis Fitness parameters Av. fecundity Hatchability (%) Ro Unsel Pop 9.67 ±0.44 B 86.35 ±1.76 A 2.78 ±0.08 A Bifen-Sel 7.13 ±0.35 C 67.85 ±2.96 B 1.61 ±0.10 B Cross1 10.43 ±0.47 AB 84.92 ±5.04 A 2.96 ±0.17 A Cross2 11.9 ±0.92 A 85.65 ±10.37 A 3.41 ±0.35 A ANOVA parameters F =11.6; P = 0.0028 F = 13.3; P = 0.0018 F = 14.4; P = 0.0014 Population
rm 0.11 ±0.00 AB 0.07 ±0.00 C 0.11 ±0.01 B 0.13 ±0.01 AB F = 22.7; P = 0.0003
The means within a column denoted by different letters are significantly different (P<0.05). df values for ANOVA = 3,8
26
Rf 1 ±0.00 B 0.58 ±0.03 C 1.07 ±0.03 AB 1.22 ±0.03 A F = 17.2; P = 0.0008
14.00
Bp and MRGR
12.00 10.00
A AB
B
8.00
C
Bp
6.00 A
4.00
C
B
B
MRGR
2.00 0.00 Unsel pop Bifen Sel Cross1 Populations
Cross2
Figure 1. Comparison of means of biotic potential (Bp) and mean relative growth rate (MRGR) among different populations of Oxycarenus hyalinipennis. The population bars with different letters are significantly different from each other (P<0.05).
27
28
Highlights • • • • •
Selection for 12 generations with bifenthrin induced 55.64-fold resistance in DCB. Relaxing from selection pressure reverted bifenthrin resistance to 24.93-fold No obvious cross-resistances were observed with all tested insecticides Bifenthrin-selected population was low in fitness. Bifenthrin resistance was incompletely dominant and autosomal
Declaration of Interest Statement All the authors declare that they have no conflict of interest.
S0261-2194(20)30040-5
DOI:
https://doi.org/10.1016/j.cropro.2020.105107
Reference:
JCRP 105107
To appear in:
Crop Protection
Received Date: 28 December 2019 Revised Date:
2 February 2020
Accepted Date: 6 February 2020
Please cite this article as: Banazeer, A., Shad, S.A., Shahzad Afzal, M.B., Laboratory induced bifenthrin resistance selection in Oxycarenus hyalinipennis (Costa) (Hemiptera: Lygaeidae): Stability, cross-resistance, dominance and effects on biological fitness, Crop Protection (2020), doi: https:// doi.org/10.1016/j.cropro.2020.105107. This is a PDF file of an article that has undergone enhancements after acceptance, such as the addition of a cover page and metadata, and formatting for readability, but it is not yet the definitive version of record. This version will undergo additional copyediting, typesetting and review before it is published in its final form, but we are providing this version to give early visibility of the article. Please note that, during the production process, errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain. © 2020 Published by Elsevier Ltd.
Credit Author Statement AB conceived the study. MBSA and SAS designed the study. AB collected the insects and carried out the research work. SAS supervised the laboratory work. MBSA provided technical guidelines to process the raw data. AB analyzed the data. AB and MBSA wrote the manuscript. SAS read and approved the written manuscript.
1.4 1.2 Relative fitness
50 40 30 20 10
1 0.8 0.6 0.4 0.2
0
0 G1
G3
G5
G7
G9
G13
UNSEL
Bifen-Sel
Generations bioassayed with bifenthrin
Cross1
Populations
0.84 Degree of dominance
Selection of bifenthrin resistance
60
0.82 0.8 0.78 0.76 0.74 0.72 0.7 Cross1
Cross2
Hybrid populations
Cross2
1
Laboratory
induced
bifenthrin
resistance
selection
in
Oxycarenus
2
hyalinipennis (Costa) (Hemiptera: Lygaeidae): stability, cross-resistance,
3
dominance and effects on biological fitness
4
Ansa Banazeer,a Sarfraz Ali Shad,a* Muhammad Babar Shahzad Afzala,b**
5
a
6
Zakariya University, Multan, Punjab, Pakistan
7
b
8
Running Title: Bifenthrin resistance selection and comparison of fitness costs
9
To whom correspondence should be addressed:
Department of Entomology, Faculty of Agricultural Sciences and Technology, Bahauddin
Citrus Research Institute, Sargodha, Punjab, Pakistan
10
*Dr. Sarfraz Ali Shad
11
Department of Entomology, Faculty of Agricultural Sciences and Technology, Bahauddin
12
Zakariya University, Multan, Pakistan
13
Email: [email protected]
14
**Dr. Muhammad Babar Shahzad Afzal
15
Department of Entomology, Faculty of Agricultural Sciences and Technology, Bahauddin
16
Zakariya University, Multan, Pakistan and Citrus Research Institute, Sargodha, Punjab, Pakistan
17
Email: [email protected]
18
ABSTRACT
19
The dusky cotton bug, Oxycarenus hyalinipennis (Costa) (Hemiptera: Lygaeidae) is a serious
20
pest of cotton and damages cotton seed by reducing the oil content. In Pakistan, O. hyalinipennis
21
is managed by using various insecticides and has developed resistance to several insecticides. In
22
this study, O. hyalinipennis was selected with bifenthrin for 12 generations (G1 to G12) and
23
developed a 55.64-fold resistance when compared with an unselected population (Unsel Pop).
24
Bifenthrin resistance declined from 55.64 to 24.93-fold when selected population was removed
25
from bifenthrin selection pressure for four generations (G13 to G16). Bifenthrin selected (Bifen-
26
Sel) population showed a very low cross-resistance to profenofos (2.82-fold), deltamethrin (2.35-
27
fold) and acetamiprid (2.21-fold) when compared with a field population (Field Pop). The
28
bifenthrin resistance was incomplelety dominanat and autosomal. The relative fitness (Rf) of
29
Bifen-Sel population was 0.58 along with significant decreases in average nymphal survival,
30
fecundity, egg hatchability, intrinsic rate of population increase (rm), net reproductive rate (Ro)
1
31
and biotic potential (Bp). The Rf of Cross1 and Cross2 was 1.07 and 1.22, respectively. The high
32
fitness costs, instable resistance and a very low cross-resistance with other insecticides might be
33
useful in slowing down the evolution of bifenthrin resistance by implementing an insecticide
34
rotation plan.
35
Keywords: Dusky cotton bug; insecticide; biological traits; cross-resistance; resistance inversion
36 37 38 39 40 41 42 43 44
2
45
1. Introduction
46
Dusky cotton bug Oxycarenus hyalinipennis (Costa) (Hemiptera: Lygaeidae) is a sucking
47
pest of cotton in Pakistan (Jaleel et al., 2014) and commonly referred as the cotton seed bug or
48
cotton stainer bug (Schaefer and Panizzi, 2000). Geographically, O. hyalinipennis is distributed
49
in Asia, Africa, Europe, Central America, the Middle East, South America and the Caribbean
50
(Halbert and Dobbs, 2010). Adult females of O. hyalinipennis lay eggs in cotton lint when bolls
51
open (Kirkpatrick, 1923). Both nymphs and adults of O. hyalinipennis feed on the seeds and suck
52
oil; excessive feeding reduces the seed weight and oil content (Schaefer and Panizzi, 2000). The
53
staining of cotton occurs during picking, and also due to crushing of bugs and their excrement in
54
ginning factories. The fiber becomes dirty due to cast skin and dead insect bodies and may
55
produce an unpleasant smell which ultimately reduces lint quality and market value (Kirkpatrick,
56
1923).
57
Insecticides are the main tools to manage O. hyalinipennis in cotton (Ullah et al., 2015).
58
Insecticides from different chemical groups such as organophosphates, pyrethroids, and
59
neonicotinoids are sprayed in Pakistan multiple times (about 8-10) in the cotton fields during the
60
season (Saeed et al., 2018). As a result of frequent insecticide applications, insecticide resistance
61
has developed in field populations of O. hyalinipennis against various insecticides (Ullah et al.,
62
2015). Studies on insecticide resistance are crucial to know the severity of resistance and to
63
devise suitable pest resistance management strategies. Bifenthrin has been commonly used for
64
both sucking and chewing pests of cotton since 1986 (Ali, 2018). However, the excessive use of
65
this insecticide has resulted in development of resistance in american bollworm Helicoverpa
66
armigera (Hübner) (Lepidoptera: Noctuidae) (Ahmad et al., 1997), spotted bollworm Earias
67
vittella (Fab.) (Lepidoptera: Noctuidae) (Ahmad and Arif, 2009; Jan et al., 2015), cotton
68
mealybug Phenacoccus solenopsis Tinsley (Homoptera: Pseudococcidae) (Saddiq et al., 2014),
69
O. hyalinipennis (Ullah et al., 2015) and cotton jassid Amrasca devastans (Dist.) (Homoptera:
70
Cicadellidae) (Abbas et al., 2018). Moreover, a few studies have reported on bifenthrin
71
resistance in laboratory-selected populations of P. solenopsis (Mansoor et al., 2016) and O.
72
hyalinipennis (Bilal et al., 2018).
73
In resistant genotypes, fitness costs associated with development of resistance are crucial
74
to study as they provide better insight into evolution of insecticide resistance (Gassmann et al.,
75
2009). The biological fitness of resistant population measured in terms of insect survival, 3
76
developmental times, fecundity and net reproductive rates is often reduced following continuous
77
laboratory selection as compared to its counterpart population in the absence of pesticides, hence
78
can be exploited to slow the pace of resistance (Roush and McKenzie, 1987). Fitness costs of
79
pyrethroid resistance have been explored previously against deltamethrin resistance in P.
80
solenopsis (Saddiq et al., 2016a) and Heliothis virescens Fabricius (Lepidoptera: Noctuidae)
81
(Sayyed et al., 2008), and lambda-cyhalothrin resistance in M. domestica (Abbas et al, 2016).
82
Previously, an O. hyalinipennis population was selected with bifenthrin to characterize activities
83
of different detoxificatin enzymes and to evaluate cross-resistance to profenofos, chlorpyrifos,
84
lambda-cyhalothrin, imidacloprid and chlorfenapyr when the selection was stopped (Bilal et al.,
85
2018). However, this study did not provide any information about fitness costs and genetics of
86
bifenthrin resistance. Morover, Bilal et al. (2018) did not study the cross-resistance spectrum to
87
profenofos, deltamethrin and acetamiprid at different selected generations.
88
In this study, we selected field collected O. hyalinipennis with bifenthrin in the laboratory
89
with the aim to verify its resistance development and also to explore cross-resistance possibilities
90
with some other insecticides by conducting bioassays at different generations of O. hyalinipennis
91
during the selection process. The resistant strain was also removed from bifenthrin selection
92
pressure for several generations to determine its impact on resistance stability. We further
93
investigated whether bifenthrin resistance was inherited by sex-linked or autosomal genes and
94
also its extent of dominance. Fitness traits linked with bifenthrin resistance in the resistant and
95
unselected populations of O. hyalinipennis and also in the progeny of their reciprocal crosses
96
were also studied. This study can help with the implementation of insecticide resistance
97
management programs against O. hyalinipennis.
98 99 100
2 Material and methods 2.1. Collection and rearing of insects
101
About 1000 nymphs and adults of O. hyalinipennis were randomly collected from highly
102
infested cotton bolls in a cotton field of the Central Cotton Research Institute (CCRI), Multan
103
(30.19°N, 71.46°E), Pakistan. The field from where the population was collected was sprayed
104
annually about 4-5 times with organophosphate, pyrethroid and neonicotinoids to control O.
105
hyalinipennis (Personal communication with scientific officer). The infested cotton bolls were
106
shaken and insects were collected in plastic jars. After collection, both nymphs and adults were 4
107
separated in the laboratory by using aspirators to obtain a homogenous population to use in
108
bioassays. The population was kept in plastic jars (24 × 12 cm) that were covered with muslin
109
clothes under constant laboratory conditions at 27±2ºC temperature, 60 ± 5% relative humidity
110
and 14:10 L:D hours. Insects were reared on open cotton bolls along with fresh branches of
111
China rose Hibiscus rosasinensis L. as their leaves also provided moisture for the insects. Old
112
branches were replaced with fresh ones twice a week and cotton bolls were changed after every
113
month. The field collected insects were reared for one generation (Go) to eliminate inherent field
114
effects and to increase the population before conducting the bioassays.
115 116
2.2. Populations After acclimatizing the population to laboratory conditions, field collected insects (Field
117
Pop) were divided in two sub-populations; one sub-population was selected with varying
118
concentrations of bifenthrin for 12 generations and designated as “Bifen-Sel” population. The
119
second sub-population was reared parallel to Bifen-Sel population without any insecticide
120
treatment and was designated as “Unsel Pop”. Two reciprocal crosses were also established by
121
crossing the naïve adult insects: 15 males from Bifen-Sel and 15 females from Unsel Pop were
122
crossed to obtain Cross1 progeny and 15 females of Bifen-Sel with 15 males of Unsel Pop were
123
crossed to get Cross2 progeny. Male and female adults were identified by their shape of abdomen
124
i.e. truncate in female and round in male. Moreover, males are shorter in size than females
125
(Henry, 1983).
126
2.3. Insecticides
127
Commercially available formulations of insecticides were used for bioassays: bifenthrin
128
(Talstar® 10EC, FMC, Pakistan), deltamethrin (Decis Super® 10EC, Bayer Crop Sciences,
129
Pakistan), profenofos (Curacron® 500EC, Syngenta, Pakistan) and acetamiprid (Mospilon® 20
130
SP, Arysta Life Sciences, Pakistan).
131
2.4. Bioassays
132
Toxicity bioassays of different insecticides were performed by using the standard IRAC
133
008 leaf-dip method (IRAC, 2019). Two-to-three days old adults of O. hyalinipennis were used
134
in each bioassay. A stock solution (40 mL) was prepared for making five serial concentrations
135
and each concentration was tested in three replicates. Fresh leaves of H. rosasinensis were
136
dipped in each concentration for ten-seconds and air dried for 1-2 hours at room temperature
137
before the insect exposure. Dried leaves were kept in Petri-dishes (5 cm in diameter) on a
5
138
slightly moistened filter paper to prevent the leaves from desiccation. A total of thirty adults
139
(both males and females) were tested for each concentration, ten in each replicate. A total of 180
140
adult insects were used in a bioassay including the control. Insects in the control were allowed to
141
feed on leaves dipped in tap water only. Mortality was assessed after 48 and 72 hours. Adults
142
were considered dead when there was no coordinated movement after a slight touch with a camel
143
hair brush.
144
2.5. Selection protocol for bifenthrin resistance
145
A preliminary bioassay of bifenthrin was performed with the Field Pop to determine the
146
lethal concentrations (LCs) required for selection of O. hyalinipennis with bifenthrin, so that
147
sufficient survivors were left for the next generation. Adults (2-3 days old) were selected with
148
bifenthrin continuously with different lethal concentrations (LC5-LC80) of insecticide ranging
149
from 17-2865 ppm from G1 to G12 (Table 1). The leaf-dip method was used for the selection
150
bioassays by using fresh leaves of H. rosasinensis. For each selection, bifenthrin treated leaves
151
after air drying were kept in Petri-dishes and adults ranging from 160-532 (30-35 per Petri-dish)
152
were exposed. The treated population in Petri-dishes were placed under laboratory conditions at
153
27±2ºC temperature, 60±5% relative humidity and 14:10 h light:dark photoperiod (Ullah et al.,
154
2016). Mortality data were taken after 48 hours exposure to bifenthrin in each selection.
155
Survivors of each selection were reared to get the next progeny for subsequent selection and an
156
average survival of 74.75% was maintained during selections.
157
2.6. Cross-resistance in bifenthrin-selected population
158
The cross-resistance ratio (CRR) in bifenthrin-selected strains was evaluated with other
159
insecticides (mentioned in the insecticides section) as compared to the field population. It was
160
calculated as follows:
161 162
CRR = LC50 of insecticides in Bifen-Sel / LC50 of insecticides in Field Pop 2.7. Stability of bifenthrin resistance
163
The bifenthrin-selected population was reared unselected in the laboratory for four
164
generations from G13-G16 to determine the resistance stability against bifenthrin and some other
165
insecticides. The bioassays with different insecticides were performed at G16 and resistance
166
ratios of Bifen-Sel (G16) were calculated by comparing LC50 of insecticide in Bifen-Sel (G16)
167
with corresponding insecticide LC50 in Unsel Pop.
168
6
169
2.8. Degree of dominance The degree of dominance (DLC) of Bifen-Sel strain was calculated by using the formula
170 171
of Bourguet et al. (2000): = (
1
2 – Unsel Pop) / ( Bifen − Sel − Unsel Pop)
172
Where X is the log of LC50 of tested strain. DLC values vary from 0-1, DLC = 0 indicates
173
completely recessive, DLC = 1 indicates completely dominant nature of insecticide resistance;
174
while DLC 0.50 to <1 and >0<0.50 indicates incompletely dominant and incompletely recessive
175
resistance, respectively.
176
2.9. Procedure to study fitness cost parameters
177
To study the fitness components, a total of 90 newly emerged 3rd instars from Bifen-Sel,
178
Unsel, Cross1 (Bifen-Sel males × Unsel females) and Cross2 (Bifen-Sel females × Unsel males)
179
populations were randomly chosen as starting individuals. Insects from each population were
180
placed in separate plastic jars (14 × 9 cm) containing open cotton bolls and twigs of H.
181
rosasinensis for feeding. The experiment was performed under laboratory conditions as
182
mentioned above. The 90 insects from each population were subdivided into three replicates (30
183
insects each). The following parameters were recorded for each population: developmental
184
duration, survival rate, fecundity and egg hatchability. The fecundity was determined as number
185
of eggs laid by single female until female died and egg hatchability was measured as number of
186
nymphs hatched from eggs. The percent egg hatching was determined as number of nymphs
187
hatched/total eggs ×100 in each replicate. The eggs unable to hatch were considered as non-
188
viable.
189 190
The mean relative growth rate (MRGR) was determined by following formula (Radford, 1967): MRGR = [W2 (mg) – W1 (mg)] / T
191 192
In this equation, W2 and W1 are weights of 1st instar and 5th instar, respectively, and T is time
193
duration from 1st to 5th instar.
194
Net reproductive rate (Ro) was calculated according to Cao and Han (2006): Ro = Nn+1 / Nn
195 196 197 198
Where Nn is the starting generation insects while Nn+1 represents next generation offspring. The Birch (1948) formula was used to calculate intrinsic rate of population increase (rm) as follows: 7
rm = Ro / DT
199 200 201
Where Ro is net reproductive rate and DT is the developmental time from egg to adult. The biotic potential (Bp) was determined using the formula of Roush and Plapp (1982):
202
Bp = [Fecundity (F) / Developmental time ratio (DTr)]
203
Where fecundity (F) was measured as average number of eggs / female and DTr was estimated
204
as DT of tested population / DT of Unsel Pop
205 206 207
The following equation was used to find the relative fitness of resistant and hybrid strains as compared to unselected strain: Relative fitness (Rf) = Ro of Bifen-Sel or Cross1 or Cross2 / Ro of Unsel Pop
208 209
2.10. Data analysis Probit software (version 1.5) was used for analysis of toxicity data (Finney, 1971).
210
Different probit parameters such as LC50, 95% Fiducial limits (FLs), slope with standard errors
211
(SEs) and chi-square (χ2) of each tested insecticide were determined by probit analysis.
212
Statistically, two LC50 values were considered similar if their 95% FLs overlapped (Litchfield
213
and Wilcoxon, 1949). The resistance and cross-resistance ratios determined were categorized as
214
very high (>100), high (51-100), medium (21-50), low (11-20), very low (2-10) and no (<2)
215
resistance (Torres-Vila et al., 2002). The 95% confidence limits (CLs) of resistance and cross-
216
resistance ratios were also determined according to method of Robertson et al. (2007) and
217
considered significant if these did not include the value of 1. The life-history data of all
218
populations were analyzed by statistix software (version 8.1) to estimate relative fitness of tested
219
populations compared to Unsel Pop using completely randomized design. The mean values of
220
different life history data of each population were compared by Least Significant Difference
221
(LSD) test at 5% of significance level.
222 223
3. Results
224
3.1. Toxicities of different insecticides at field and unselected populations and resistance status
225
of field population of O. hyalinipennis
226
For Field Pop, toxicities of profenofos (LC50 = 94.03 ppm) and deltamethrin (LC50 =
227
59.65 ppm) were similar (95% FLs overlapped) but higher (95% FLs did not overlap) than that
228
of bifenthrin (LC50 = 504.2 ppm). Moreover, acetamiprid toxicity (LC50 = 13.6 ppm) was higher
229
(95% FLs did not overlap) than all other insecticides tested for Field Pop. All the insecticides 8
230
had higher toxicities (95% FLs did not overlap) to Unsel Pop than those for Field Pop. The
231
resistance ratios of Field Pop for profenofos, bifenthrin, deltamethrin and acetamiprid were 3.34-
232
, 4.97-, 2.96- and 2.48-fold, respectively, compared to Unsel Pop (Table 2).
233 234 235
3.2. Toxicity and resistance development to bifenthrin in O. hyalinipennis and resistance reversion The LC50s of Bifen-Sel at G3, G5, G7, G9 and G13 using bifenthrin were 890.25, 969.75,
236
1622.94, 4787.03, and 5649.18 ppm, respectively. The Bifen-Sel population of O. hyalinipennis
237
had resistance ratios of 1.77-, 1.92-, 3.21-, 9.49- and 11.20-fold against bifenthrin compared with
238
the Field Pop when tested at the above mentioned generations (Table 2). However, the selected
239
strains showed resistance ratios of 8.77-, 9.55-, 15.98-, 47.14- and 55.64-fold at G3, G5, G7, G9,
240
and G13, respectively, as compared to the Unsel Pop (Table 2). The Bifen-Sel (G13), after rearing
241
for four generations without exposure to bifenthrin, when tested using profenofos, bifenthrin,
242
deltamethrin, and acetamiprid at G16 exhibted resistance ratios of 5.44-, 24.93-, 3.86-, and 4.76-
243
fold, respectively as compared to Unsel Pop. However, toxicities of profenofos, bifenthrin,
244
deltamethrin, and acetamiprid were similar between Bifen-Sel (G16) and Bifen-Sel (G13) for each
245
of these four insecticides (95% FLS overlapped) (Table 2).
246
3.3. Pattern of cross-resistance to Bifen-Sel in O. hyalinipennis
247
The Bifen-Sel population of O. hyalinipennis was evaluated for cross-resistance to
248
profenofos, deltamethrin and acetamiprid at G3, G5, G7, G9, G11 and G13 compared with the Field
249
Pop. The Bifen-Sel population showed no to very low cross-resistance to profenofos (0.77-2.82-
250
fold), deltamethrin (0.85-2.35-fold) and acetamiprid (1.03-2.21-fold) as the selection progressed
251
from G1 to G12 (Table 3).
252
3.4. Resistance and degree of dominance of reciprocal populations
253
The Cross1 and Cross2 populations showed 5.39- and 9.42-fold resistance to bifenthrin,
254
respectively, in comparison to the Unsel Pop. The DLC values of bifenthrin resistance for Cross1
255
and Cross2 were 0.74 and 0.82, respectively indicating the incomplete dominant inheritance of
256
resistance. Moreover, toxicities of bifenthrin were similar (95% FLs overlapped) when tested
257
against both hybrid populations of O. hyalinipennis (Table 4).
258
9
259
3.5. Fitness parameters for Unsel, Bifen-Sel, Cross1 and Cross2 in O. hyalinipennis
260
3.5.1. Developmental durations
261
Mean development durations of different stages in Unsel, Bifen-Sel, Cross1 and Cross2
262
are summarized in Table 5. The developmental duration of eggs did not differ among all tested
263
populations. Developmental duration from 1st to 5th instar in Bifen-Sel was similar to Unsel Pop
264
but significantly higher in Cross1 and Cross2. The adult male longevity of Bifen-Sel, Cross1, and
265
Cross2 was similar to Unsel Pop but it was higher in Cross2 as compared to Cross1. The adult
266
female was significantly higher in Cross2 than that in the other three populations.
267
3.5.2. Survival rates
268
The survival rate of 1st and 2nd instars was significantly lower in Bifen-Sel but similar in
269
Cross1 and Cross2 as compared to Unsel Pop (Table 5). The survival rate of 3rd instar was
270
significantly lower in Bifen-Sel and Cross1 than that of Unsel Pop. No significant differences
271
were found in the survival of 4th and 5th instars in all tested populations. The average survival rate
272
from 1st to 5th instar was significantly lower in Bifen-Sel compared to the other three
273
populations.
274
3.5.3. Fecundity, egg hatchability, net reproductive rate (Ro), intrinsic rate of population
275
increase (rm) and relative fitness (Rf)
276
The Bifen-Sel females produced significantly lower number of eggs compared with the
277
Unsel Pop, Cross1 and Cross2. The Unsel pop had a significantly lower fecundity than Cross2 but
278
not than Cross1. The percent egg hatchability and net reproductive rate (Ro) in the Bifen-Sel
279
were significantly less when compared with the other three populations. The intrinsic rate of
280
population increase (rm) was significantly lower in Bifen-Sel compared to all other populations
281
which had similar rm. The relative fitness (Rf) of Bifen-Sel was significantly lower than that of
282
the other three populations. Cross1 had a similar Rf than Unsel Pop. Cross2 had a significantly
283
higher Rf than Unsel Pop (Table 6).
284
3.5.4. Biotic potential (Bp) and mean relative growth Rate (MRGR)
285
The Bp of Bifen-Sel was significantly lower in comparison with the Unsel Pop. However
286
hybrid populations showed similar Bp to Unsel Pop (F = 15.7; P = 0.0010; df = 3,8) (Fig 1). A
287
significantly higher MRGR was found in all the tested populations as compared to Unsel Pop (F
288
= 52.8; P<0.0001; df = 3,8) (Fig 1).
289
10
290
4. Discussion
291
In the current study, selection of O. hyalinipennis for 12 generations resulted in a
292
substantial increase in LC50 and resistance ratios to bifenthrin. The LC50 of bifenthrin in the Field
293
Pop at G1 was 504.2 ppm with very low resistance (4.97-fold) before selection in the laboratory.
294
However, when the same population was continuously selected until G12, the LC50 of bifenthrin
295
in Bifen-Sel population (G13) increased to 5649.18 ppm or 55.64-fold compared with the Unsel
296
Pop. Laboratory induced high to very high levels of resistance selections have been reported in
297
the house fly Musca domestica Linnaeus (Diptera: Muscidae) to beta-cypermethrin with 4420-
298
fold resistance after 25 selected generations compared with the susceptible strain (Zhang et al.,
299
2008), 474-fold resistance to cypermethrin in H. armigera over six selected generations
300
compared with the laboratory population (Achaleke and Brévault, 2010), 236-fold resistance to
301
deltamethrin in Plutella xylostella (L.) (Lepidoptera: Plutellidae) after six selected generations
302
compared with the unselected population (Sayyed et al., 2005), and 54.32-fold and 178.42-fold
303
resistance to bifenthrin in P. solenopsis after 14 generations when compared with the field and
304
laboratory strains, respectively (Mansoor et al., 2016). The resistance levels selected under
305
laboratory conditions can be low or high depending upon the selection pressure, previous history
306
of exposure to insecticides, insect species, selection protocols, geographic origins of populations
307
and presence of susceptible and resistant genes.
308
In this study, the decline in bifenthrin resistance from 55.64-fold to 24.93-fold (about
309
55% decrease) after a few non-selected generations with the Bifen-Sel population suggested the
310
unstable nature of bifenthrin resistance in O. hyalinipennis. Similar to our results, reversion of
311
insecticide resistance has been reported previously in O. hyalinipennis after five unexposed
312
generations against chlorfenapyr (Ullah and Shad, 2017) and bifenthrin (Bilal et al., 2018).
313
Recovery of resistant populations towards susceptibility could be due to high fitness costs
314
associated with insecticide resistance (Gassmann et al., 2009) which are in agreement with the
315
present findings.
316
.The Bifen-Sel population of O. hyalinipennis did not show any cross-resistance to
317
profenofos and deltamethrin up to the fifth selected generation. However, as the selection
318
continued to the 12th generation, there was a slight increase (very low) in cross-resistance against
319
both profenofos (2.01-2.82-fold), and deltamethrin (2.00-2.35-fold) when bioassayed at G13.
320
Similarly, the Bifen-Sel population did not have any cross-resistance to acetamiprid until at G7 11
321
but a very low cross-resistance (2.35-, 2.11-, and 2.21-fold) was expressed at G9, G11, and G13.
322
Lack of and/or very low cross-resistance in the Bifen-Sel population against profenofos and
323
acetamiprid was expected as both of these insecticides have entirely different mode of actions
324
and belong to different chemical groups than bifenthrin. Profenofos is an organophosphorus
325
insecticide and is known to act as acetylcholinesterase (AChE) inhibitor at synapse while
326
acetamiprid is neonicotinoid group insecticide and works as nicotinic acetylcholine receptor
327
(nAChR) competitive modulator. However, bifenthrin is a pyrethroid insecticide and targets the
328
axon of neuron as sodium channel modulator (IRAC, 2019). Unexpectedly, presence of very low
329
cross-resistance in the Bifen-Sel population of O. hyalinipennis against another pyrethroid
330
(deltamethrin) might be due to limited exposure of the pest population to deltamethrin in the
331
field and involvement of several resistance mechanisms. A deltamethrin-selected population
332
(4632.8-fold) of P. solenopsis had no cross-resistance to profenofos but a low level of cross-
333
resistance against lambda-cyhalothrin and acetamiprid (Afzal et al., 2018). Results of this study
334
are similar to Mansoor et al. (2016) who found negligible cross-resistance in a bifenthrin selected
335
population of P. solenopsis against buprofezin, chlorpyrifos and lambda-cyhalothrin.
336
Insect populations often experience a decline in biological fitness during the development
337
of resistance. These phenomena have been found in many insecticide resistance studies such as
338
in P. xylostella to Bacillus thuringiensis toxin Cry2Ad (Liao et al., 2019), brown planthopper
339
Nilaparvata lugens (Stål) (Hemiptera: Delphacidae) to nitenpyram (Zhang et al., 2018), M.
340
domestica to spinosad (Khan, 2018), Spodoptera litura (Fab.) (Lepidoptera: Noctuidae) to
341
bistrifluron (Huang et al., 2019), fall armyworm Spodoptera frugiperda (J.E. Smith)
342
(Lepidoptera: Noctuidae) to Bt toxin Cry1A.105 (Niu et al., 2018), green peach aphid Myzus
343
persicae (Sulzer) (Hemiptera: Aphididae) to sulfoxaflor (Wang et al., 2018), H. armigera to
344
indoxacarb (Cui et al., 2018), western flower thrips Frankliniella occidentalis (Thysanoptera:
345
Thripidae) and maize armyworm Mythimna separata (Lepidoptera: Noctuidae) to thiamethoxam
346
(Gao et al., 2014; Yasoob et al., 2018), and P. solenopsis to spirotetramat (Ejaz and Shad, 2017).
347
In this study, bifenthrin resistance development in the Bifen-Sel population resulted in declines
348
in nymphal survival, fecundity, egg hatchability, Ro, rm, and Rf (0.58) as compared with Unsel
349
Pop. These findings indicate that, if selection discontinued, the Bifen-Sel population might not
350
increase as rapidly as the Unsel Pop because of significant disadvantages in life history
351
parameters. These results also suggest the presence of a trade-off in distribution of energy 12
352
resources between costs in the form of fitness and mechanism(s) of resistance. Similar to these
353
results, lower relative fitness due to insecticide resistance has been reported in other insects
354
under different selection regimes such as in yellow fever mosquito Aedes aegypti (Linnaeus)
355
(Diptera: Culicidae) (Jaramillo-O et al., 2014) and M. domestica (Abbas et al., 2016) to lambda-
356
cyhalothrin resistance; and P. solenopsis to deltamethrin resistance (Saddiq et al., 2016a).
357
Determination of fitness costs associated with resistance is important to study in
358
homozygous resistant individuals but also in heterozygotes that can act as a carrier of most
359
abundant resistant genes in early stages of resistance (Jia et al., 2009). Owing to this, fitness
360
costs of bifenthrin resistance were also estimated in two hybrid populations (Cross1 and Cross2)
361
of Bifen-Sel and Unsel Pop. The results showed that the Rf of Cross1 (1.07) and Cross2 (1.22)
362
were similar and both population progenies had superior fitness traits when compared with the
363
the parental Bifen-Sel. Moreover, in both hybrids, fitness recovery was significantly higher in
364
Cross2 compared with the Unsel Pop. The advantageous biological characteristics such as
365
survival, fecundity, egg hatchability, Ro and rm in reciprocal cross progenies compared to Bifen-
366
Sel population might be attributed to their increased vigor, or heterosis, which might have
367
resulted in longer inbreeding leading to the improvement of beneficial traits in heterozygous
368
progeny. Therefore, the study of life history parameters of both resistant and reciprocal
369
individuals is crucial and helpful in formulating resistance management strategies (Jia et al.,
370
2009).
371
The hybrid progenies with resistance ratios of 5.39-fold (Cross1) and 9.42-fold (Cross2)
372
were also used to determine inheritance characteristics associated with bifenthrin resistance. The
373
overlapping of FLs of LC50 values in both populations suggested that maternal effects were not
374
involved in bifenthrin resistance in O. hyalinipennis and it was autosomal. Moreover, DLC of
375
both populations was >0.50 and <1, which indicated that bifenthrin resistance was incompletely
376
dominant. Autosomal and incompletely dominant resistance have also been reported in many
377
insect pests (Sayyed et al., 2005; Achaleke and Brévault, 2010; Khan et al., 2015; Saddiq et al.
378
2016b; Ullah et al., 2016; Ijaz and Shad, 2018).
379
5. Conclusion and recommendations
380
The present findings showed that O. hyalinipennis has a high potential to develop
381
bifenthrin resistance following continuous selection pressure. The high resistance was also
382
reverted upon removal of selection pressure and no obvious cross-resistance occured with any 13
383
tested insecticide. Moreover, fitness costs were apparent in the resistant population. Bifenthrin
384
resistance inheritance appeared to be autosomal and incompletely dominant. The unstable
385
bifenthrin resistance indicated that evolution of resistance to bifenthrin may be minimized in O.
386
hyalinipennis if the insecticide is removed from a management program for a given duration. A
387
very weak cross-resistance, high fitness costs and incomplete dominance of bifenthrin resistance
388
suggests that these insecticides can be included in a rotation program to delay resistance
389
development. Recovery of fitness traits in hybrid populations also indicated that mixing the
390
selected (resistant) and unselected populations might suppress the resistance by diluting the
391
resistant genes.
392 393
14
394 395 396
Conflict of interest statement No potential conflict of interest exists among all authors Author contributions
397
AB conceived the study. MBSA and SAS designed the study. AB collected the insects
398
and carried out the research work. SAS supervised the laboratory work. MBSA provided
399
technical guidelines to process the raw data. AB analyzed the data. AB and MBSA wrote the
400
manuscript. SAS read and approved the written manuscript.
401 402
Acknowledgements We acknowledge Prof. Dr. José Eduardo Serrão, Department of General
403
Federal University of Viçosa, Brazil, and Prof. (Retd.) Dr. Muhammad Aslam, Department of
404
Entomology, Faculty of Agricultural Sciences and Technology, Bahauddin Zakariya University,
405
Multan, Pakistan for sparing their time to check manuscript for improvement of English
406
language and sense.
407
15
Biology,
408
References
409
Abbas, N., Ismail, M., Ejaz, M., Asghar, I., Irum, A., Shad, S.A., Binyameen, M., 2018.
410
Assessment of field evolved resistance to some broad-spectrum insecticides in cotton
411
jassid, Amrasca devastans from southern Punjab, Pakistan. Phytoparasitica 46, 411–419.
412
Abbas, N., Shah, R.M., Shad, S.A., Iqbal, N., Razaq, M., 2016. Biological trait analysis and
413
stability of lambda-cyhalothrin resistance in the house fly, Musca domestica L. (Diptera:
414
Muscidae). Parasitol. Res. 115, 2073–2080.
415
Achaleke, J., Brévault, T., 2010. Inheritance and stability of pyrethroid resistance in the cotton
416
bollworm Helicoverpa armigera (Lepidoptera: Noctuidae) in Central Africa. Pest
417
Manage. Sci. 66, 137–141.
418 419
Afzal MBS, Shad SA, 2015. Resistance inheritance and mechanism to emamectin benzoate in Phenacoccus solenopsis (Homoptera: Pseudococcidae). Crop Prot. 71, 60–65
420
Afzal, M.B.S., Shad, S.A., Ejaz, M., Ijaz, M., 2018. Selection, cross-resistance, and resistance
421
risk assessment to deltamethrin in laboratory selected Phenacoccus solenopsis
422
(Homoptera: Pseudococcidae). Crop Prot. 112, 67–73.
423
Ahmad, M., Arif, M.I., 2009. Resistance of Pakistani field populations of spotted bollworm
424
Earias vittella (Lepidoptera: Noctuidae) to pyrethroid, organophosphorus and new
425
chemical insecticides. Pest Manage. Sci. 65, 433–439.
426 427
Ahmad, M., Arif, M.I., Attique, M.R., 1997. Pyrethroid resistance of Helicoverpa armigera (Lepidoptera: Noctuidae) in Pakistan. Bull. Entomol. Res. 87, 343–347.
428
Ali, M.A., 2018. The pesticides registered with recommendations for safe handling and use in
429
pakistan, a hand book for agriculture extension agents. Published by Directorate General
430
Agriculture (Extension & AR) Punjab, Lahore.
431
Bilal, M., Freed, S., Ashraf, M.Z., Rehan, A., 2018. Resistance and detoxification enzyme
432
activities to bifenthrin in Oxycarenus hyalinipennis (Hemiptera: Lygaeidae). Crop Prot.
433
111, 17–22.
434 435 436 437
Birch, L., 1948. The intrinsic rate of natural increase of an insect population. J. Anim. Ecol. 17, 15–26. Bourguet, D., Genissel, A., Raymond, M., 2000. Insecticide resistance and dominance levels. J. Econ. Entomol. 93, 1588–1595.
16
438 439 440
Cao, G., Han, Z., 2006. Tebufenozide resistance selected in Plutella xylostella and its cross‐resistance and fitness cost. Pest Manage. Sci. 62, 746–751. Cui, L., Wang, Q., Qi, H., Wang, Q., Yuan, H., Rui, C., 2018. Resistance selection of indoxacarb
441
in
Helicoverpa
armigera
(Hübner)
(Lepidoptera:
Noctuidae):
cross‐resistance,
442
biochemical mechanisms and associated fitness costs. Pest Manage. Sci. 74, 2636–2644.
443
Ejaz, M., Shad, S.A, 2017. Spirotetramat resistance selected in the Phenacoccus solenopsis
444
(Homoptera: Pseudococcidae): cross-resistance patterns, stability, and fitness costs
445
analysis. J. Econ. Entomol. 110, 1226–1234.
446
Finney, D.J., 1971. Probit Analysis, third ed. Cambridge University Press, UK, 333 pp.
447
Gao, C.F., Ma, S.Z., Shan, C.H., Wu, S.F., 2014. Thiamethoxam resistance selected in the
448
western flower thrips Frankliniella occidentalis (Thysanoptera: Thripidae): Cross-
449
resistance patterns, possible biochemical mechanisms and fitness costs analysis. Pestic.
450
Biochem. Physiol. 114, 90–96.
451 452
Gassmann, A.J., Carrière, Y., Tabashnik, B.E., 2009. Fitness costs of insect resistance to Bacillus thuringiensis. Ann. Rev. Entomol. 54, 147–163.
453
Halbert, S.E., Dobbs, T., 2010. Cotton Seed Bug, Oxycarenus hyalinipennis (Costa): A serious
454
pest of cotton that has become established in the Caribbean Basin. FDACS-Pest Alert
455
DACS-P-01726, Florida Department of Agriculture and Consumer Services, Division of
456
Plant Industry [Online] http://www. freshfromflorida. com/pi/pest_alerts/pdf/cotton-
457
seedbug-pest-alert. pdf. Accessed 6, 2012.
458 459
Henry, T.J., 1983. Pests not known to occur in the United States or of limited distribution, USDA-APHIS-PPQ, APHIS 43, 1–6.
460
Huang, Q., Wang, X., Yao, X., Gong, C., Shen, L., 2019. Effects of bistrifluron resistance on the
461
biological traits of Spodoptera litura (Fab.) (Noctuidae: Lepidoptera). Ecotoxicol. 28,
462
323–332.
463
Ijaz, M., Shad, S.A., 2018. Inheritance mode and realized heritability of resistance to
464
imidacloprid in Oxycarenus hyalinipennis Costa (Hemiptera: Lygaeidae). Crop Prot. 112,
465
90–95.
466
IRAC, 2019. IRAC Mode of Action Classification Scheme.
17
467
Jaleel, W., Saeed, S., Naqqash, M.N., Zaka, S.M., 2014. Survey of Bt cotton in Punjab Pakistan
468
related to the knowledge, perception and practices of farmers regarding insect pests. Int.
469
J. Agric. Crop Sci. 7, 10–20.
470
Jan, M.T., Abbas, N., Shad, S.A., Saleem, M.A., 2015. Resistance to organophosphate,
471
pyrethroid and biorational insecticides in populations of spotted bollworm, Earias vittella
472
(Fabricius) (Lepidoptera: Noctuidae), in Pakistan. Crop Prot. 78, 247–252.
473
Jaramillo-O. N., Fonseca-Gonzalez, I., Chaverra-Rodrıguez, D., 2014. Geometric morphometrics
474
of nine field isolates of Aedes aegypti with different resistance levels to lambda-
475
cyhalothrin and relative fitness of one artificially selected for resistance. PLoS ONE 9,
476
96379.
477
Jia, B., Liu, Y., Zhu, Y.C., Liu, X., Gao, C., Shen, J., 2009. Inheritance, fitness cost and
478
mechanism of resistance to tebufenozide in Spodoptera exigua (Hübner) (Lepidoptera:
479
Noctuidae). Pest Manag. Sci. 65, 996–1002.
480
Khan, H. A.A., Akram, W., Haider, M.S. 2015. Genetics and mechanism of resistance to
481
deltamethrin in the house fly, Musca domestica L., from Pakistan. Ecotoxicol. 24, 1213–
482
1220.
483 484 485 486
Khan, H.A.A., 2018. Spinosad resistance affects biological parameters of Musca domestica Linnaeus. Sci. Rep. 8, 14031. Kirkpatrick, T., 1923. The Egyptian Cotton Seed Bug. Ministry Agricult. Egypt., Techn. a. Sci. Serv. Bull. 35, 1–106.
487
Liao, J., Xue, Y., Xiao, G., Xie, M., Huang, S., You, S., Wyckhuys, K.A., You, M., 2019.
488
Inheritance and fitness costs of resistance to Bacillus thuringiensis toxin Cry2Ad in
489
laboratory strains of the diamondback moth, Plutella xylostella (L.). Sci. Rep. 9, 6113.
490 491
Litchfield, J.T., Wilcoxon, F., 1949. A simplified method of evaluating dose–effect experiments. J. Pharmacol. Exp. Theory 99, 99–103.
492
Mansoor, M.M., Afzal, M.B.S., Basoalto, E., Raza, A.B.M., Banazeer, A., 2016. Selection of
493
bifenthrin resistance in cotton mealybug Phenacoccus solenopsis Tinsley (Homoptera:
494
Pseudococcidae):
495
mechanism. Crop Prot. 87, 55–59.
496 497
Cross-resistance,
realized
heritability and
possible
resistance
Niu, Y., Head, G.P., Price, P.A., Huang, F., 2018. Inheritance and fitness costs of Cry1A. 105 resistance in two strains of Spodoptera frugiperda (JE Smith). Crop Prot. 110, 229–235. 18
498
Radford, P.J., 1967. Growth Analysis Formulae-Their Use and Abuse 1. Crop Sci. 7, 171–175.
499
Robertson, J.L., Savin, N., Preisler, H.K., Russell, R.M., 2007. Bioassays with Arthropods,
500 501 502 503 504
second ed. CRC press, Boca Raton, Florida. Roush, R.T., McKenzie, J.A., 1987. Ecological genetics of insecticide and acaricide resistance. Ann. Rev. Entomol. 32, 361–380. Roush, R.T., Plapp, F.W., 1982. Effects of insecticide resistance on biotic potential of the house fly (Diptera: Muscidae). J. Econ. Entomol. 75, 708–713.
505
Saddiq, B., Abbas, N., Shad, S.A., Aslam, M., Afzal, M.B.S., 2016a. Deltamethrin resistance in
506
the cotton mealybug, Phenacoccus solenopsis Tinsley: cross-resistance to other
507
insecticides, fitness cost analysis and realized heritability. Phytoparasitica 44, 83–90.
508
Saddiq, B., Afzal, M.B.S., Shad, S.A. 2016b. Studies on genetics, stability and possible
509
mechanism of deltamethrin resistance in Phenacoccus solenopsis Tinsley (Homoptera:
510
Pseudococcidae) from Pakistan. J.Genet. 95, 1009–1016.
511
Saddiq, B., Shad, S.A., Khan, H.A.A., Aslam, M., Ejaz, M., Afzal, M.B.S., 2014. Resistance in
512
the mealybug Phenacoccus solenopsis Tinsley (Homoptera: Pseudococcidae) in Pakistan
513
to selected organophosphate and pyrethroid insecticides. Crop Prot. 66, 29–33.
514
Saeed, R., Abbas, N., Razaq, M., Mahmood, Z., Naveed, M., Rehman, H.M.U., 2018. Field
515
evolved resistance to pyrethroids, neonicotinoids and biopesticides in Dysdercus koenigii
516
(Hemiptera: Pyrrhocoridae) from Punjab, Pakistan. Chemosphere 213, 149–155.
517
Sayyed, A.H., Ahmad, M., Crickmore, N., 2008. Fitness costs limit the development of
518
resistance to indoxacarb and deltamethrin in Heliothis virescens (Lepidoptera:
519
Noctuidae). J. Econ. Entomol. 101, 1927–1933.
520
Sayyed, A.H., Attique, M.N.R., Khaliq, A., Wright, D.J., 2005. Inheritance of resistance and
521
cross‐resistance to deltamethrin in Plutella xylostella (Lepidoptera: Plutellidae) from
522
Pakistan. Pest Manage. Sci. 61, 636–642.
523 524 525 526
Schaefer, C.W., Panizzi, A.R., 2000. Economic importance of Heteroptera: a general view, Heteroptera of economic importance. CRC Press, pp. 25-30. Schaefer, C.W., Panizzi, A.R., 2000. Economic importance of Heteroptera: a general view, Heteroptera of economic importance. CRC Press, pp. 25–30.
527 528
19
529
Torres-Vila, L. M., Rodrı́guez-Molina, M. C., Lacasa-Plasencia, A., Bielza-Lino, P., Rodrı́guez-
530
del-Rincón, Á., 2002. Pyrethroid resistance of Helicoverpa armigera in Spain: current
531
status and agroecological perspective. Agric. Ecosys. Environ. 93, 55–66.
532
Ullah, S., Shad, S.A., Abbas, N., 2015. Resistance of dusky cotton bug, Oxycarenus
533
hyalinipennis Costa (Lygaidae: Hemiptera), to conventional and novel chemistry
534
insecticides. J. Econ. Entomol. 109, 345–351.
535
Ullah, S., Shah, R.M., Shad, S.A., 2016. Genetics, realized heritability and possible mechanism
536
of chlorfenapyr resistance in Oxycarenus hyalinipennis (Lygaeidae: Hemiptera). Pestic.
537
Biochem. Physiol. 133, 91–96.
538
Ullah, S., Shad, S.A., 2017. Toxicity of insecticides, cross-resistance and stability of
539
chlorfenapyr resistance in different strains of Oxycarenus hyalinipennis Costa
540
(Hemiptera: Lygaeidae). Crop Prot. 99, 132–136.
541
Wang, Z.H., Gong, Y.J., Chen, J.C., Su, X.C., Cao, L.J., Hoffmann, A.A., Wei, S.J., 2018.
542
Laboratory selection for resistance to sulfoxaflor and fitness costs in the green peach
543
aphid Myzus persicae. J. Asia-Pacific Entomol. 21, 408–412.
544
Yasoob, H., Abbas, N., Li, Y., Zhang, Y., 2018. Selection for resistance, life history traits and
545
the biochemical mechanism of resistance to thiamethoxam in the maize armyworm,
546
Mythimna separata (Lepidoptera: Noctuidae). Phytoparasitica 46, 627–634.
547 548
Zhang, L., Shi, J., Gao, X., 2008. Inheritance of beta‐cypermethrin resistance in the housefly Musca domestica (Diptera: Muscidae). Pest Manage. Sci. 64, 185–190.
549
Zhang, X., Mao, K., Liao, X., He, B., Jin, R., Tang, T., Wan, H., Li, J., 2018. Fitness cost of
550
nitenpyram resistance in the brown planthopper Nilaparvata lugens. J. Pest Sci. 91,
551
1145–1151.
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552
Table 1. History of selection of Oxycarenus hyalinipennis with bifenthrin in the laboratory Generation G1 G2 G3 G4 G5 G6 G7 G8 G9 G10 G11 G12
Concentration (ppm) 17 35 59 172 504.21 504.21 653.55 850.67 1,000 1324.2 2029.4 2865
No exposed 400 370 524 532 407 200 160 220 180 300 281 411
No died 45 46 27 17 273 60 64 39 30 40 91 221
No survived 355 324 497 515 134 140 96 181 150 260 190 190
553
21
%Mortality 11.25 12.43 5.43 3.20 67.08 30.00 40.00 17.73 16.67 13.33 32.38 53.77
%Survival 88.75 87.57 94.85 96.80 32.92 70.00 60.00 82.27 83.33 86.67 67.62 46.23
Table 2. Response of different populations of Oxycarenus hyalinipennis to tested insecticides
a
Population
Insecticide
LC50 (ppm)
95% FL (ppm)
Slope(±S.E)
χ2
DF
P
Na
RRb(95%CL)
RRc(95%CL)
Field Pop (G1)
Profenofos
94.03
70.88-125.91
1.89(±0.29)
0.24
4
0.99
180
1
3.34(2.24-4.98)
Field Pop (G1)
Bifenthrin
504.2
355.018-852.924
1.12(±0.20)
1.2
4
0.88
180
1
4.97(3.00-8.21)
Field Pop (G1)
Deltamethrin
59.65
41.18-98.71
1.34(±0.27)
2.79
4
0.59
180
1
2.96(1.76-4.98)
Field Pop (G1)
Acetamiprid
13.6
9.00-28.84
1.48(±0.30)
0.67
4
0.95
180
1
2.48(1.38-4.48)
Unsel Pop
Profenofos
28.14
21.11-38.07
1.84(±0.29)
1.85
4
0.76
180
1
1
Unsel Pop
Bifenthrin
101.54
75.83-138.77
1.79(±0.29)
1.5
4
0.83
180
1
1
Unsel Pop
Deltamethrin
20.17
14.86-29.69
1.68(±0.28)
2.32
4
0.68
180
1
1
Unsel Pop
Acetamiprid
5.48
4.13-7.35
1.88(±0.29)
1.55
4
0.82
180
1
1
Bifen-Sel (G3)
Bifenthrin
890.25
626.93-1394.28
1.42(±0.27)
1
4
0.91
180
1.77(1.11-3.18)
8.77(4.65-12.26)
Bifen-Sel (G5)
Bifenthrin
969.75
710.99-1430.41
1.64(±0.29)
1.15
4
0.89
180
1.92(1.13-3.26)
9.55(6.13-14.89)
Bifen-Sel (G7)
Bifenthrin
1622.94
1143.40-2698.50
1.46(±0.28)
0.72
4
0.95
180
3.21(1.82-5.70)
15.98(9.76-26.18)
Bifen-Sel (G9)
Bifenthrin
4787.03
3062.35-11650.08
1.44(±0.31)
0.31
4
0.99
180
9.49
47.14(24.57-90.46)
Bifen-Sel (G13)
Bifenthrin
5649.18
3680.06-12389.88
1.27(±0.28)
0.67
4
0.95
180
11.20(5.72-21.95)
55.64(30.29-102.19)
Bifen-Sel (G13)
Profenofos
265.1
187.33-450.10
1.53(±0.29)
1.17
4
0.88
180
2.82(1.73-4.61)
9.42(5.75-15.45)
Bifen-Sel (G13)
Deltamethrin
140.01
102.53-221.17
1.78(±0.31)
0.57
4
0.97
180
2.35(1.37-4.03)
6.94(4.27-11.30)
Bifen-Sel (G13)
Acetamiprid
30.04
21.84-47.13
1.65(±0.30)
1.94
4
0.75
180
2.21(1.17-4.16)
5.48(3.48-8.66)
Bifen-Sel (G16)
Profenofos
153.02
106.91-275.02
1.58(±0.30)
0.75
4
0.83
180
-
-
Bifen-Sel (G16)
Bifenthrin
2531.55
1663.38-5578.06
1.45(±0.30)
0.33
4
0.99
180
-
-
Bifen-Sel (G16)
Deltamethrin
77.91
54.18-139.41
1.63(±0.30)
1.29
4
0.86
180
-
-
Bifen-Sel (G16)
Acetamiprid
26.1
18.47-45.37
1.49(±0.29)
1.26
4
0.87
180
-
-
Number of adult insects treated in each bioassay including control Resistance ratio = LC50 of insecticides in selected population / LC50 of insecticides in Field Pop c Resistance ratio = LC50 of insecticides in Field or Bifen-Sel population (G3 to G13) / LC50 of insecticides in Unsel Pop d Resistance ratio = LC50 of insecticide in Bifen-Sel population (G16) / LC50 of insecticide in Unsel Pop b
22
RRd(95%CL)
5.44(3.24-9.12) 24.93(13.55-45.90) 3.86(2.25-6.64) 4.76(2.90-7.85)
Table 3. Cross-resistance evaluation against some insecticides at different generations of Bifen-Sel population of Oxycarenus hyalinipennis Population Insecticide LC50 (ppm) 95% FL (ppm) Slope (±S.E) χ2 DF P Na CRRb(95%CL) Field Pop (G1) Profenofos 94.03 70.88-125.91 1.89(±0.29) 0.24 4 0.99 180 Field Pop (G1) Deltamethrin 59.65 41.18-98.71 1.34(±0.27) 2.79 4 0.59 180 Field Pop (G1) Acetamiprid 13.6 9.00-28.84 1.48(±0.30) 0.67 4 0.95 180 74.25 54.02-99.065 1.81(±0.29) 3.40 4 0.49 180 0.79(0.53-1.18) Bifen-Sel (G3) Profenofos Bifen-Sel (G5) Profenofos 72.85 50.37-100.69 1.57(±0.28) 1.10 4 0.89 180 0.77(0.50-1.20) Bifen-Sel (G7) Profenofos 189.19 139.91-283.94 1.74(±0.30) 0.24 4 0.99 180 2.01(1.30-3.11) Bifen-Sel (G9) Profenofos 195.74 147.20- 275.24 1.85(±0.30) 0.45 4 0.98 180 2.08(1.38-3.14) Bifen-Sel (G11) Profenofos 244.21 175.44-393.77 1.51(±0.29) 1.15 4 0.89 180 2.60(1.63-4.15) Bifen-Sel (G13) Profenofos 265.1 187.33-450.10 1.53(±0.29) 1.17 4 0.88 180 2.82(1.73-4.61) Bifen-Sel (G3) Deltamethrin 50.73 37.79-71.26 1.75(±0.29) 0.61 4 0.96 180 0.85(0.51-1.41) Bifen-Sel (G5) Deltamethrin 58.37 43.32-83.45 1.72(±0.29) 0.69 4 0.95 180 0.98(0.59-1.63) Bifen-Sel (G7) Deltamethrin 142.90 86.38- 422.80 1.12(±0.27) 0.4 4 0.98 180 2.40(1.10-5.21) Bifen-Sel (G9) Deltamethrin 152.84 94.76-406.57 1.18(±0.28 0.92 4 0.92 180 2.56(1.22-5.37) Bifen-Sel (G11) Deltamethrin 119.34 85.07-198.25 1.57(±0.29) 0.04 4 1.00 180 2.00(1.14-3.52) Bifen-Sel (G13) Deltamethrin 140.01 102.53-221.17 1.78(±0.31) 0.57 4 0.97 180 2.35(1.37-4.03) Bifen-Sel (G3) Acetamiprid 14.74 9.86-30.68 1.51(±0.31) 0.59 4 0.96 180 1.08(0.52-2.24) Bifen-Sel (G5) Acetamiprid 14.04 9.52-28 1.54(±0.31) 0.63 4 0.96 180 1.03(0.51-2.10) Bifen-Sel (G7) Acetamiprid 17 12.63-25.39 1.79(±0.30) 1.27 4 0.87 180 1.25(0.68-2.32) Bifen-Sel (G9) Acetamiprid 31.95 21.07-68.05 1.35(±0.29) 0.13 4 1.00 180 2.35(1.13-4.90) Bifen-Sel (G11) Acetamiprid 28.67 20.69-45.00 1.62(±0.29) 1.45 4 0.84 180 2.11(1.11-4.00) Bifen-Sel (G13) Acetamiprid 30.04 21.84-47.13 1.65(±0.30) 1.94 4 0.75 180 2.21(1.17-4.16) a Number of adult insects treated in each bioassay including control b Cross-resistance ratio = LC50 of each insecticide in Bifen-Sel / LC50 of corresponding insecticide in Field Pop
23
Table 4. Bifenthrin resistance and its dominance in reciprocal crosses DF P Na RRb(95% CL) DLCc Population LC50 (ppm) 95 % FL (ppm) Slope±SE χ2 Unsel Pop 101.54 75.83-138.77 1.79±0.29 1.50 4 0.83 180 1 Cross1 547.14 404.45-750.09 1.74±0.29 1.04 4 0.90 180 5.39(3.55-8.18) 0.74 956.10 704.39-1448.1 1.72±0.30 1.19 4 0.88 180 9.42(6.02-14.74) 0.82 Cross2 a Number of adult insects treated in each bioassay including control b Resistance ratio = LC50 of bifenthrin in Cross1 or Cross2 / LC50 of bifenthrin in Unsel Pop c Degree of dominance
24
Table 5. Developmental durations in days and survival rates (%) of different stages in different tested populations of Oxycarenus hyalinipennis Mean ±SE Unsel Pop Bifen-Sel Cross1 Cross2 Egg duration 5.16 ±0.47 A 5.06 ±0.89 A 5.27 ±0.18 A 5.51 ±0.37 A st th 1 to 5 instar duration 18.52 ±0.25 B 19.67 ±0.56 AB 21.15 ±0.31 A 20.05 ±0.63 A Egg to adult duration 23.68 ±0.32B 24.73 ±0.48 AB 26.420 ±0.42 A 25.56 ±1.00 AB Adult male longevity 4.8 ±0.29 AB 5.92 ±0.36 AB 4.44 ±0.80 B 6.11 ±0.11 A Adult female longevity 11.31 ±0.72 B 11.22 ±0.97 B 13.33 ±1.20 B 17.36 ±0.81 A 1st instar survival 82.03 ±2.34 A 73.28 ±1.46 B 84.85 ±1.32 A 84.75 ±1.84 A nd 2 instar survival 85.95 ±2.64 A 76.20 ±2.73 B 83.10 ±1.58 AB 80.99 ±1.76 AB rd 3 instar survival 97.78 ±1.11 A 91.11 ±2.22 B 90 ±1.92 B 94.44 ±1.11 AB th 4 instar survival 98.89 ±1.11 A 96.34 ±2.06 A 96.29 ±0.08 A 98.85 ±1.15 A th 5 instar survival 100 ±0.00 A 97.62 ±2.38 A 98.72 ±1.28 A 96.43 ±0.00 A Av. instar survival 92.87 ±0.18 A 86.91 ±0.75 B 90.59 ±1.01 A 91.09 ±0.58 A The means within a row denoted by similar letters are not significantly different (P>0.05). df values for ANOVA = 3,8 Biological traits
25
ANOVA parameters F = 0.13; P = 0.9416 F = 5.47; P = 0.0243 F = 3.63; P = 0.0645 F = 3.08; P = 0.0901 F = 9.27; P = 0.0055 F = 9.34; P = 0.0054 F = 3.38; P = 0.0746 F = 4.44; P = 0.0407 F = 1.28; P = 0.3466 F = 1.27; P = 0.3473 F = 9.68; P = 0.0049
Table 6. Comparison of means ±SE of fecundity, hatchability, Ro, rm, and and Rf of Unsel, Bifen-Sel, Cross1 and Cross2 populations of Oxycarenus hyalinipennis Fitness parameters Av. fecundity Hatchability (%) Ro Unsel Pop 9.67 ±0.44 B 86.35 ±1.76 A 2.78 ±0.08 A Bifen-Sel 7.13 ±0.35 C 67.85 ±2.96 B 1.61 ±0.10 B Cross1 10.43 ±0.47 AB 84.92 ±5.04 A 2.96 ±0.17 A Cross2 11.9 ±0.92 A 85.65 ±10.37 A 3.41 ±0.35 A ANOVA parameters F =11.6; P = 0.0028 F = 13.3; P = 0.0018 F = 14.4; P = 0.0014 Population
rm 0.11 ±0.00 AB 0.07 ±0.00 C 0.11 ±0.01 B 0.13 ±0.01 AB F = 22.7; P = 0.0003
The means within a column denoted by different letters are significantly different (P<0.05). df values for ANOVA = 3,8
26
Rf 1 ±0.00 B 0.58 ±0.03 C 1.07 ±0.03 AB 1.22 ±0.03 A F = 17.2; P = 0.0008
14.00
Bp and MRGR
12.00 10.00
A AB
B
8.00
C
Bp
6.00 A
4.00
C
B
B
MRGR
2.00 0.00 Unsel pop Bifen Sel Cross1 Populations
Cross2
Figure 1. Comparison of means of biotic potential (Bp) and mean relative growth rate (MRGR) among different populations of Oxycarenus hyalinipennis. The population bars with different letters are significantly different from each other (P<0.05).
27
28
Highlights • • • • •
Selection for 12 generations with bifenthrin induced 55.64-fold resistance in DCB. Relaxing from selection pressure reverted bifenthrin resistance to 24.93-fold No obvious cross-resistances were observed with all tested insecticides Bifenthrin-selected population was low in fitness. Bifenthrin resistance was incompletely dominant and autosomal
Declaration of Interest Statement All the authors declare that they have no conflict of interest.