Development of alpha-cypermethrin resistance and its effect on biological parameters of yellow fever mosquito, Aedes aegypti (L.) (Diptera: Culicidae).

Alpha-cypermethrin interacts with the sodium channel and causes nerve blockage in insects. It is used to manage Aedes aegypti (Linnaeus) (Diptera: Culicidae), a primary vector of dengue worldwide. It not only affects both target and non-target organisms, but overuse of this insecticide increases the chances of resistance development in insect pests. In this study, resistance development, biological parameters, and stability of alpha-cypermethrin resistance were studied in a laboratory-selected strain of Ae. aegypti. The alpha-cypermethrin selected strain (Alpha Sel) developed an 11.86-fold resistance level after 12 rounds of alpha-cypermethrin selection compared to the unselected strain (Unsel). In biological parameters, Alpha Sel and Cross1 (Unsel ♂ and Alpha Sel♀) had shorter larval durations compared to Unsel and Cross2 (Unsel ♀ and Alpha Sel ♂) populations. The pupal duration of Alpha Sel and both crosses was shorter than that in the Unsel strain. The relative fitness of Alpha Sel, Cross1, and Cross2 was significantly less than that of the Unsel strain. These results indicate that alpha-cypermethrin resistance comes with fitness costs. Moreover, the frequency of alpha-cypermethrin resistance decreased when the Alpha Sel population was reared without further selection pressure for four generations. So, resistance was unstable and reversed when insecticide pressure ceased. We concluded that the judicious and rotational use of different insecticides with different modes of action and the adoption of other IPM-recommended practices would suppress resistance development for more extended periods in Ae. aegypti.

Risk assessment of cyromazine and methoxyfenozide resistance suggests higher additive genetic but lower environmental variation supporting quick resistance development in non-target Chrysoperla carnea (Stephens).

Insecticides are effective against economic pests, but these pose serious threats to the environment and ecosystem components such as natural enemies. Resistance risk assessment forecasts insecticide resistance development in target pests and non-target biological control agents under special conditions. Field-collected Chrysoperla carnea was selected with two Insect Growth Regulators (IGRs) viz. cyromazine and methoxyfenozide for 15 generations to determine the resistance development potential of this natural enemy. Selection to cyromazine and methoxyfenozide induced 759.08-fold and 3531.67-fold resistance with realized heritability of 0.37 and 0.62 in C. carnea, respectively, suggesting higher additive genetic variations in first half of selection (h2 = 0.46 for cyromazine and h2 = 0.75 for methoxyfenozide) than in second half (h2 = 0.18 and 0.25, respectively). Estimates of projected rate of resistance development indicate C. carnea will take only 6 to 2 generations at h2 = 0.37, 8 to 2 at h2 = 0.27, and 5 to 2 at h2 = 0.27, at constant slope = 1.81 for a tenfold increase in cyromazine resistance. At h2 = 0.37, 3-1, and 10-8 generations would be needed for this increase in LC50 if slope = 0.82 and 2.82, respectively. Similarly, it may take 3 to 1 generations at h2 = 0.62 and 0.72, but 4 to 1 at h2 = 0.52, at constant slope = 1.62, for a tenfold increase in methoxyfenozide resistance. On the same h2 = 0.62, 1-0, and 5-1 generations would be required for increase if slope = 0.62 and 2.62, respectively. Selection and resistance to both insecticides induced an insignificant difference in the sex ratio of C. carnea. These results confirm that this natural enemy has tremendous potential for resistance development under selection pressure.

Genetics and mechanism of resistance to chlorantraniliprole in Musca domestica L. (Diptera: Muscidae).

The house fly, Musca domestica L. is an important mechanical vector of different pathogens of medical and veterinary importance. It is an organism well-known for its ability to develop insecticide resistance. In the current study, we investigated the genetic basis and mechanism of chlorantraniliprole resistance in a field strain of house fly by selecting it artificially in the laboratory with a commercial formulation of chlorantraniliprole (CTPR-SEL). After seven generations of consecutive selection with chlorantraniliprole, CTPR-SEL strain developed a 644-fold resistance compared with the Susceptible strain and a 3-fold resistance compared with the field strain. Reciprocal crossing between the CTPR-SEL and Susceptible homozygous strains revealed an autosomal and incomplete dominant mode of resistance to chlorantraniliprole. A direct test using a monogenic inheritance model based on chi-square analysis revealed that the resistance was governed by more than one gene. Bioassays with synergists indicated that esterases might be involved in the resistance of house fly to chlorantraniliprole. These findings may be helpful to the development of an improved strategy for chlorantraniliprole resistance management in house fly.

Fitness cost, realized heritability and stability of resistance to spiromesifen in house fly, Musca domestica L. (Diptera: Muscidae).

The house fly, Musca domestica L. (Diptera: Muscidae), is an insect pest of public health and veterinary importance. Spiromesifen is a new chemistry insecticide widely used for the management of sucking insect pests of vegetables and crops. In the present study, assessment of resistance risk and fitness costs associated with spiromesifen resistance in M. domestica was studied. Moreover, stability of resistance to spiromesifen and other tested insecticides (fipronil, spinosad, and bifenthrin) was evaluated in the spiromesifen-selected-strain (SPIRO-SEL-POP). After 7-generations of selection with spiromesifen, SPIRO-SEL-POP developed 108.76-fold resistance compared with the unselected strain (UNSEL-POP). The estimated value of realized heritability was 0.59 for spiromesifen resistance. Due to withdrawal of spiromesifen selection for five generations (F6-F10) on SPIRO-SEL-POP, a decline in LC50 values against spiromesifen, spinosad and bifenthrin was 0.16, 0.14 and 0.13-folds, respectively. In biological trait experiments, larval weight of Cross1 (SPIRO-SEL-POP♀ × UNSEL-POP♂) and SPIRO-SEL-POP was significantly lower than that of Cross2 (SPIRO-SEL-POP♂ × UNSEL-POP♀) and UNSEL-POP. Pupal weight of SPIRO-SEL-POP was higher when compared with Cross1 while it was similar to that of Cross2 and UNSEL-POP. Adult emergence rate of UNSEL-POP was higher than Cross1, but similar to the Cross2 and SPIRO-SEL-POP. The SPIRO-SEL-POP and Cross1 showed the lowest relative fitness when compared with USEL-POP and Cross2. Intrinsic rate of natural increase of SPIRO-SEL-POP was much lower than that of UNSEL-POP and Cross2 followed by Cross1. The SPIRO-SEL-POP exhibited lower biotic potential when compared with UNSEL-POP and Cross2 but similar to Cross1. Fecundity and hatching rates were lower in SPIRO-SEL-POP compared to UNSEL-POP. It could be concluded that spiromesifen resistance in M. domestica comes with a cost and is instable. Therefore, spiromesifen rotation with other insecticides and withdraw of its usage for some period could help to sustain its efficacy by delaying the development of resistance.

Characterization of inheritance and preliminary biochemical mechanisms of spirotetramat resistance in Phenacoccus solenopsis Tinsley: An economic pest from Pakistan.

Phenacoccus solenopsis is an economically important insect pest of different agronomic and horticultural field crops. In Pakistan, the cotton crop was severely attacked by P. solenopsis during 2007 and since then a varied group of insecticides are used by farmers to manage this pest. As a result, insecticide resistance has become a barrier in control of P. solenopsis. The current study was designed to explore the basics of genetics, realized heritability and possible genetic mechanisms of resistance against spirotetramat in P. solenopsis. Before selection, the wild population (Wild-Pop) showed 5.97-fold resistance when compared with lab-reared susceptible strain (Susceptible Lab-Pop). The P. solenopsis was selected with spirotetramat to 21 generations, called Spiro-SEL Pop, which showed 463.21-fold resistance as compared with the Susceptible Lab-Pop. The values of LC50 for F1 (Spiro-SEL Pop ♂ × Susceptible Lab-Pop ♀) and F1 (Spiro-SEL Pop ♀ × Susceptible Lab-Pop ♂) populations were statistically similar and values of dominance level were 0.42 and 0.54, respectively. Reciprocal crosses between Susceptible Lab-Pop and Spiro-SEL Pop showed that resistance was of autosomal in nature with incomplete dominant traits. According to the fit test, monogenic model estimation of the number of genes, which are responsible for the development of spirotetramat resistance in a population of P. solenopsis, showed that multiple genes are involved in controlling the resistance levels in tested strains of P. solenopsis. The value of heritability for resistance against spirotetramat was 0.13 in P. solenopsis. Our results suggested the presence of a metabolic-based resistance mechanism associated with the monooxygenases in P. solenopsis, while testing the synergism mechanism. These results will provide the baseline to design an effective control strategy to manage P. solenopsis in the field.

Assessing the combined toxicity of conventional and newer insecticides on the cotton mealybug Phenacoccus solenopsis.

Reduced susceptibility to insecticides often results in failure of insect pest control and repetitive use of broad-spectrum insecticides, which could have detrimental effects on beneficial arthropods and surrounding agro-ecosystems. The cotton mealybug, Phenacoccus solenopsis Tinsley is a pest of worldwide importance that can be effectively controlled using a number of insecticides. This insect has developed resistance due to injudicious use and repeated exposure to insecticides throughout the year. The aim of the present study was to investigate the toxicity of the insecticides chlorpyrifos, deltamethrin, spinosad, emamectin benzoate and indoxacarb tested either singly or in combination on laboratory susceptible (Lab-PK) and field population (Field Pop) of P. solenopsis. In the Field Pop, combination of chlorpyrifos with either spinosad or emamectin benzoate or indoxacarb showed a synergistic effect at 1:1, 1:10 and 1:20. The combination of deltamethrin with either emamectin benzoate or indoxacarb at all ratios showed synergistic effect, while the combination of deltamethrin with spinosad at 1:1 and 1:20 ratios showed antagonistic effects and that at 1:10 ratio showed synergistic effect against Field Pop. The results of synergism experiments using piperonyl butoxide (PBO) and S,S,S-tributylphosphorotrithioate (DEF) enzyme inhibitors, when combined with insecticides against the Field Pop, toxicities of all tested insecticides were significantly increased. The present study suggests that insecticide combinations and synergism could help achieve effective control of less susceptible populations of P. solenopsis. This would contribute to suitably managing resistance of this pest to insecticides with reduced negative impacts on the surrounding environment.

Laboratory selection of chlorpyrifos resistance in an Invasive Pest, Phenacoccus solenopsis (Homoptera: Pseudococcidae): Cross-resistance, stability and fitness cost.

The cotton mealybug, Phenacoccus solenopsis is an important polyphagous sucking pest of ornamentals, horticultural and fiber crops worldwide. Some P. solenopsis populations have developed insecticide resistance. This study evaluated cross-resistance, stability of insecticide resistance and life history traits affected by chlorpyrifos resistance in P. solenopsis. After nine generations selected with chlorpyrifos, P. solenopsis exhibited a 539.76-fold resistance level compared to an unselected population (UNSEL Pop). Chlorpyrifos selected population (Chlor-SEL Pop) displayed moderate cross-resistance to profenofos, nitenpyram and high cross-resistance to lambda-cyhalothrin. Biological parameters of P. Solenopsis were affected by chlorpyrifos resistance. The Chlor-SEL Pop had a significant reduction in fitness (relative fitness=0.10), along with significant decreases in pupal weight, fecundity, egg hatching %, intrinsic rate of natural population increase, biotic potential, and mean relative growth rate. It is concluded that selection with chlorpyrifos had marked effect on resistance development in P. solenopsis and upon removal of selection pressure chlorpyrifos resistance declined significantly indicating unstable resistance. Development of resistance led to high fitness costs for the chlorpyrifos-selected strain. These findings should be helpful for better and more successful resistance management of P. solenopsis.

Biological trait analysis and stability of lambda-cyhalothrin resistance in the house fly, Musca domestica L. (Diptera: Muscidae).

House flies, Musca domestica L., (Diptera: Muscidae), are pests of poultry and have the ability to develop resistance to insecticides. To design a strategy for resistance management, life history traits based on laboratory observations were established for lambda-cyhalothrin-resistant, susceptible and reciprocal crosses of M. domestica strains. Bioassay results showed that the lambda-cyhalothrin-selected strain developed a resistance ratio of 98.34 compared to its susceptible strain. The lambda-cyhalothrin-selected strain had a relative fitness of 0.26 and lower fecundity, hatchability, lower number of next generation larvae, and net reproductive rate compared with its susceptible strain. Mean population growth rates, such as intrinsic rate of population increase, and biotic potential were lower for the lambda-cyhalothrin-selected strain compared to its susceptible strain. Resistance to lambda-cyhalothrin, indoxacarb, and abamectin was unstable while resistance to bifenthrin and methomyl was stable in the lambda-cyhalothrin-selected strain of M. domestica. Development of resistance can cost considerable fitness for the lambda-cyhalothrin-selected strain. The present study provided useful information for making potential management strategies to delay resistance development in M. domestica.

Effects of different animal manures on attraction and reproductive behaviors of common house fly, Musca domestica L.

Insects rely mainly on their well-developed and highly sophisticated olfactory system to discriminate volatile cues released from host and nonhost substances, mates, oviposition substrates, and food sources. Onset of first mating, mating duration, and onset of first oviposition, oviposition period, fecundity (number of eggs laid by a female), and longevity of freshly emerged Musca domestica L. (Diptera: Muscidae) adults were observed in the presence of different animal manures: cow, horse, donkey, poultry, and an artificial diet. The M. domestica adults exposed to horse manure showed a delay in onset of first mating and first oviposition, prolonged mating duration, and reduced fecundity compared to the artificial diet (control). Likewise, the fecundity was reduced in the presence of donkey manure as compared to artificial diet. The onset of first mating was delayed and duration of first mating was shortened in the presence of cow manure as compared to artificial diet and no oviposition was observed throughout the duration of the experiment. However, the reproductive behaviors and all fitness measures in adults exposed to poultry manure were similar or even better, compared to the artificial diet. Surprisingly, in a free-choice attraction assay, the highest numbers of adult flies were attracted toward the cow manure as compared to all other manures as well as the artificial diet. However, the numbers of flies captured in all other types of manures were not different than the artificial diet (control). Furthermore, chemical analysis of headspace samples of manures revealed qualitative differences in odor (volatile) profiles of all manures and artificial diet, indicating that behavioral differences could be due to the differences in the volatile chemistry of the adult ovipositional substrates and larval growth mediums. This study may contribute toward both understanding the linkage between ecological adaptations and host selection mechanisms and the development of pest management strategies against this serious pest of medical and veterinary importance.

Toxicity of 25 synthetic insecticides to the field population of Culex quinquefasciatus Say.

The Culex quinquefaciatus Say, commonly known as the southern house mosquito, is well known for biting nuisance and vectoring of some fatal diseases. Synthetic chemicals have been relied upon as the major control measure to control mosquitoes. Therefore, we have evaluated 21 insecticides belonging to different chemical classes for their toxicity to C. quinquefaciatus females. Chlorfenapyr was the most toxic adulticide among all the tested insecticides. Among pyrethroids, deltamethrin was the least toxic adulticide, and all other have same toxicity. In case of organophosphates, the chlorpyrifos was the most toxic insecticide. Neonicotinoids such as acetamiprid, nitenpyram, and clothianidin have similar toxicity based on overlapping of 95 % confidence intervals (CI) and were more toxic when compared with the imidacloprid. The spinetoram was more toxic as compared with the spinosad (based on non-overlapping 95 % Cl). In case of ketoenoles, spirotetrament was more toxic as compared with the spiromesifen. Emamectin benzoate was the most toxic insecticide when compared with fipronil and indoxacarb. We also have tested four insect growth regulators (IGRS) including lufenuron, methoxyfenozide, pyriproxyfen, and cyromazine as larvicides. The lufenuron and pyriproxyfen have similar toxicity based upon their overlapping 95 % CI and were more toxic as compared with the methoxyfenozide and cyromazine. The methoxyfenozide was the moderately toxic among all the tested IGRS, and cyromazine was the least toxic among all the tested IGRS. These results will prove helpful in effectuating an effective integrated vector management program for C. quinquefaciatus.

Genetics, realized heritability and possible mechanism of chlorfenapyr resistance in Oxycarenus hyalinipennis (Lygaeidae: Hemiptera).

Dusky cotton bug (DCB), Oxycarenus hyalinipennis (Lygaeidae: Hemiptera) is a serious pest of cotton and other malvaceous plants. Chlorfenapyr, a broad spectrum, N-substituted, halogenated pyrrole insecticide is used extensively to control many insect pests in cotton, including DCB. In this study, we investigated a field strain of DCB to assess its potential to develop resistance to chlorfenapyr. After six generations of continuous selection pressure with chlorfenapyr, DCB had a 7.24-fold and 149.06-fold resistance ratio (RR) at G1 and G6, respectively. The genetic basis of inheritance of chlorfenapyr resistance was also studied by crossing the chlorfenapyr selected (Chlorfenapyr-SEL) and laboratory population (Lab-PK). Results revealed an autosomal and incompletely dominant mode of inheritance for chlorfenapyr resistance in the Chlorfenapyr-SEL population of DCB. The results of the monogenic model test showed chlorfenapyr resistance was controlled by multiple genes. Estimated realized heritability for chlorfenapyr resistance in the tested DCB strain was 0.123. Synergism bioassays with piperonyl butoxide and S, S, S-butyl phosphorotrithioate revealed chlorfenapyr resistance might be due to esterase activity. These results would be useful for devising an effective resistance management strategy against DCB.

Resistance of Dusky Cotton Bug, Oxycarenus hyalinipennis Costa (Lygaidae: Hemiptera), to Conventional and Novel Chemistry Insecticides.

The dusky cotton bug, Oxycarenus hyalinipennis Costa (Lygaidae: Hemiptera), is polyphagous in nature and has become one of the severe sucking pests of cotton in Pakistan. O. hyalinipennis has the potential to develop resistance to a number of insecticides, and as a result, O. hyalinipennis outbreaks occur. There is no previous study from Pakistan regarding O. hyalinipennis resistance to insecticides. Therefore, the aim of this study was to assess the resistance of different field populations of O. hyalinipennis to conventional (bifenthrin, deltamethrin, lambda-cyhalothrin, profenofos, triazophos) and novel chemistry (emamectin benzoate, spinosad, chlorfenapyr, imidacloprid, and nitenpyram) insecticides. Five populations of O. hyalinipennis, collected from Multan, Khanewal, Muzaffargarh, Lodhran, and Bahawalpur, were tested for resistance to selected insecticides by the leaf dip method. For three pyrethroids, the resistance ratios were in the range of 14- to 30-fold for bifenthrin, 2.14- to 8.41-fold for deltamethrin, and 9.12- to 16-fold for lambda-cyhalothrin, compared with the laboratory susceptible strain (Lab-PK). For two organophosphates, the range of resistance ratios was 12- to 14-fold for profenofos and 9.04- to 15-fold for triazophos. For five novel chemistry insecticides, the range of resistance ratios was 4.68- to 9.83-fold for emamectin benzoate, 6.38- to 17-fold for spinosad, 16- to 46-fold for chlorfenapyr, 11- to 22-fold for imidacloprid, and 1.32- to 11-fold for nitenpyram. Regular assessment of resistance to insecticides and integrated management plans like judicious use of insecticides and rotation of insecticides along with different modes of action are required to delay resistance development in O. hyalinipennis.

Genetics, realized heritability and preliminary mechanism of spinosad resistance in Phenacoccus solenopsis Tinsley (Homoptera: Pseudococcidae): an invasive pest from Pakistan.

The cotton mealybug, Phenacoccus solenopsis Tinsley (Homoptera: Pseudococcidae) has gained recognition as a key pest due to its invasive nature throughout the world. The P. solenopsis has a wide range of host plants and damages the cotton crop in various parts of the world. In view of the economic importance of this pest, a study on selection, inheritance and mechanism of spinosad resistance was conducted on P. solenopsis. Selection of field collected P. solenopsis for seven generations with spinosad resulted in a high resistance ratio of 282.45-fold. Genetic studies of spinosad resistance in P. solenopsis indicated that maternal effects are not involved in spinosad resistance; and resistance development is an autosomal and incompletely dominant trait. The number of genes involved in spinosad resistance was determined to be more than one, suggesting that resistance is controlled by multiple loci. The realized heritability (h (2)) value for spinosad resistance was 0.94. Synergism bioassays of spinosad with piperonyl butoxide and S,S,S-tributyl phosphorotrithioate showed that spinosad resistance in P. solenopsis could be due to esterase only. The study provides the basic information for implementation of effective resistance management strategies to control P. solenopsis.

Assessment of resistance risk to lambda-cyhalothrin and cross-resistance to four other insecticides in the house fly, Musca domestica L. (Diptera: Muscidae).

Lambda-cyhalothrin, a sodium channel modulator insecticide, has been used frequently for the control of house flies, Musca domestica L. (Diptera: Muscidae) worldwide, including Pakistan. This experiment was performed to determine the selection and assessment of lambda-cyhalothrin resistance evolution along with four other insecticides. After 26 generations of selection, the lambda-cyhalothrin-selected population developed 445-fold resistance to lambda-cyhalothrin compared to the susceptible population. There was low cross-resistance to bifenthrin and very low cross-resistance to methomyl, imidacloprid, and fipronil in the lambda-cyhalothrin-selected population compared to the field population (G1). Realized heritability (h (2)) of resistance to lambda-cyhalothrin, bifenthrin, methomyl, imidacloprid, and fipronil was 0.07, 0.05, 0.01, 0.08, and 0.08, respectively. The projected rate of resistance development revealed that if 90 % house flies were selected, then a tenfold increase in lethal concentration 50 occurred after 17, 20, 159, 13, and 14 generations for lambda-cyhalothrin (h (2) = 0.07, slope = 2.09), bifenthrin (h (2) = 0.05, slope = 1.73), methomyl (h (2) = 0.01, slope = 2.52), imidacloprid (h (2) = 0.08, slope = 1.89), and fipronil (h (2) = 0.08, slope = 2.03), respectively. The results of our study concluded that the house fly has the potential to develop multiple insecticide resistances following continued selection pressure with lambda-cyhalothrin. This study will be helpful for assisting the development of resistance management strategies.

Cross-resistance, stability, and fitness cost of resistance to imidacloprid in Musca domestica L., (Diptera: Muscidae).

Imidacloprid, a neonicotinoid insecticide, has been used frequently for the management of Musca domestica L., (Diptera: Muscidae) worldwide. To design the strategy for resistance management, life history traits were established for imidacloprid-resistant, susceptible counterpart, and reciprocal crosses M. domestica strains based on laboratory observations. Bioassay results showed that the imidacloprid-selected strain developed a resistance ratio of 106-fold to imidacloprid, 19-fold to nitenpyram, 29-fold to chlorpyrifos, and 3.8-fold to cypermethrin compared to that of the susceptible counterpart strain. The imidacloprid-selected strain showed very low cross-resistance against nitenpyram and cypermethrin and a lack of cross-resistance to chlorpyrifos. Resistance to imidacloprid, nitenpyram, and chlorpyrifos was unstable, while resistance to cypermethrin was stable in Imida-SEL strain of M. domestica. The imidacloprid-selected strain had a relative fitness of 0.61 and lower fecundity, hatchability, number of next-generation larvae, and net reproductive rate compared with the susceptible counterpart strain. Mean population growth rates, such as intrinsic rate of population increase and biotic potential, were lower for the imidacloprid-selected strain compared with the susceptible counterpart strain. Development of resistance can cost considerable fitness for the imidacloprid-selected strain. The present study provided useful information for making potential management strategies to overcome development of resistance.

Selection, resistance risk assessment, and reversion toward susceptibility of pyriproxyfen in Musca domestica L.

Pyriproxyfen, a juvenile hormone mimic, is an effective larvicide against many pests of veterinary and public health importance. Pyriproxyfen is a biorational insecticide having many environmentally friendly attributes that make it compatible with integrated pest management programs. This experiment was performed for the assessment of resistance evolution and reversion toward susceptibility of Musca domestica to pyriproxyfen. Repeated selection at successive generations resulted in 5.09- and 130-fold increase in lethal concentration 50 (LC50) compared to field and susceptible strain, respectively. A significant decline after 22 generations without selection suggesting resistance to pyriproxyfen was unstable in M. domestica. Realized heritability (h (2)) of resistance to pyriproxyfen was 0.035 in pyriproxyfen-selected strain of M. domestica. The projected rate of resistance development indicated that, if slope = 1.28 and h (2) = 0.035, then 46-21 generations are required for 10-fold increase in LC50 at 50-90 % selection intensity. These findings suggest that a risk for resistance development to pyriproxyfen occurred in M. domestica under continuous selection pressure. Pyriproxyfen susceptibility reversed when its application is ceased for a specified duration.

Mechanism, stability and fitness cost of resistance to pyriproxyfen in the house fly, Musca domestica L. (Diptera: Muscidae).

Pyriproxyfen, a bio-rational insecticide, used worldwide for the management of many insect pests including the house fly, Musca domestica. To devise a retrospective resistance management strategy, biological parameters of pyriproxyfen resistant (Pyri-SEL), unselected (UNSEL), Cross1 and Cross2M. domestica strains were studied in the laboratory. Additionally, the stability and mechanism of resistance was also investigated. After 30 generations of pyriproxyfen selection, a field-collected strain developed 206-fold resistance compared with susceptible strain. Synergists such as piperonyl butoxide and S,S,S-tributylphosphorotrithioate did not alter the LC50 values, suggesting another cause of target site resistance to pyriproxyfen in the Pyri-SEL strain. The resistance to all tested insecticides was unstable in Pyri-SEL strain. The relative fitness of 0.51 with lower fecundity, hatchability, lower number of next generation larvae, reduced mean population growth rate and net reproductive rate were observed in the Pyri-SEL strain compared with the UNSEL strain. The cost of fitness associated with pyriproxyfen resistance was evident in Pyri-SEL strain. The present study provides useful information for making pro-active resistance management strategies to delay resistance development.

Inheritance, realized heritability and biochemical mechanism of acetamiprid resistance in the cotton mealybug, Phenacoccus solenopsis Tinsley (Homoptera: Pseudococcidae).

The cotton mealybug, Phenacoccus solenopsis Tinsley (Homoptera: Pseudococcidae) is a serious pest in many countries of the world because of its polyphagous nature and has caused huge losses to the cotton crop. The aim of present study was to explore the mode of inheritance and mechanism of acetamiprid resistance in P. solenopsis. After five rounds of selection with acetamiprid, P. solenopsis developed a 315-fold resistance compared with the laboratory susceptible population. The LC50 values of progenies of both reciprocal crosses (F1 and F1') showed no significant difference and degree of dominance values were 0.56 and 0.93 for F1 and F1', respectively. Monogenic model of inheritance and Lande's method revealed that more than one factors were involved in acetamiprid resistance. Realized heritability (h(2)) value was 0.58 for acetamiprid resistance. A synergism study of piperonyl butoxide (PBO) and S,S,S-tributylphosphorotrithioate (DEF) with acetamiprid also showed the significant presence of P-450 mono-oxygenase and esterase in the acetamiprid resistance. Hence, acetamiprid resistance in the P. solenopsis was autosomal, incompletely dominant and polygenic. These results are a source of basic information to design and plan fruitful management programmes to control P. solenopsis.

Genetics and preliminary mechanism of chlorpyrifos resistance in Phenacoccus solenopsis Tinsley (Homoptera: Pseudococcidae).

Cotton mealybug, Phenacoccus solenopsis Tinsley, is a serious pest of cotton and other crops and infestation by this pest results in yield losses that affect the economy of Pakistan. Various groups of insecticides have been used to control this pest but resistance development is a major factor that inhibits its control in the field. Chlorpyrifos is a common insecticide used against many pests including P. solenopsis. The present experiment was designed to assess the genetics and mechanism of chlorpyrifos resistance and to develop a better resistance management strategy and assess the genetics and mechanism of chlorpyrifos resistance. Before selection, the field strain showed 3.1-fold resistance compared to the susceptible strain (CSS). After 8 rounds of selection with chlorpyrifos, a selected population developed a 191.0-fold resistance compared to the CSS. The LC50 values of F1 (CRR ♀ × CSS ♂) and F1(†) (CRR ♂ × CSS ♀) strains were not significantly different and dominance (DLC) values were 0.42 and 0.55. Reciprocal crosses between chlorpyrifos susceptible and resistant strains indicated that resistance was autosomal and incompletely recessive. The monogenic model of fit test and calculation of number of genes segregating in the chlorpyrifos resistant strain demonstrated that resistance is controlled by multiple genes. A value of 0.59 was calculated for realized heritability for chlorpyrifos resistance. Synergism bioassays with piperonyl butoxide and S, S, S-butyl phosphorotrithioate showed that chlorpyrifos resistance was associated with microsomal oxidases and esterases. It was concluded that chlorpyrifos resistance in P. solenopsis was autosomally inherited, incompletely recessive and polygenic. These findings would be helpful to improve the management of P. solenopsis.

Cross-resistance, genetics, and realized heritability of resistance to fipronil in the house fly, Musca domestica (Diptera: Muscidae): a potential vector for disease transmission.

Houseflies, Musca domestica (L.), are ubiquitous pests that have the potential to spread a variety of pathogens to humans, poultries, and dairies. Pesticides are commonly used for the management of this pest. Fipronil is a GABA-gated chloride channel-inhibiting insecticide that has been commonly used for the management of different pests including M. domestica throughout the world. Many pests have developed resistance to this insecticide. A field-collected strain of M. domestica was selected with fipronil for continuous 11 generations to assess the cross-resistance, genetics, and realized heritability for designing a resistance management strategy. Laboratory bioassays were performed using the feeding method of mixing insecticide concentrations with 20% sugar solutions and cotton soaks dipped in insecticide solutions were provided to tested adult flies. Bioassay results at G12 showed that the fipronil-selected strain developed a resistance ratio of 140-fold compared to the susceptible strain. Synergism bioassay with piperonyl butoxide (PBO) and S,S,S,-tributyl phosphorotrithioate (DEF) indicated that fipronil resistance was associated with microsomal oxidase and also esterase. Reciprocal crosses between resistant and susceptible strains showed an autosomal and incompletely dominant resistance to fipronil. The LC50 values of F1 and F'1 strains were not significantly different and dominance values were 0.74 and 0.64, respectively. The resistance to fipronil was completely recessive (D(ML) = 0.00) at the highest dose and incompletely dominant at the lowest dose (D(ML) = 0.87). The monogenic resistance based on chi-square goodness of fit test and calculation of the minimum number of segregating genes showed that resistance to fipronil is controlled by multiple genes. The fipronil resistance strain confirmed very low cross-resistance to emamectin benzoate and spinosad while no cross-resistance to chlorpyrifos and acetamiprid when compared to that of the field population. The heritability values were 0.112, 0.075, 0.084, 0.008, and 0.052 for fipronil, emamectin benzoate, spinosad, acetamiprid, and chlorpyrifos, respectively. It was concluded that fipronil resistance in M. domestica was autosomally inherited, incompletely dominant, and polygenic. These findings would be helpful for the better and successful management of M. domestica.