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.

Laboratory selection, cross-resistance, and estimations of realized heritability of indoxacarb resistance in Phenacoccus solenopsis (Homoptera: Pseudococcidae).

BACKGROUND: Cotton mealybug Phenacoccus solenopsis is a pest of cotton, vegetables, ornamentals, and medicinal plants. In many parts of the world P. solenopsis has been managed by integration of cultural, mechanical, biological and chemical methods, but in Pakistan the use of insecticide sprays has resulted in the development of resistance to some insecticides. In this study indoxacarb resistance was investigated by selecting a P. solenopsis population under laboratory conditions for many generations. The cross-resistance potential of indoxacarb resistance with other chemistries and the realized heritability of indoxacarb resistance were also evaluated. RESULTS: A field population of P. solenopsis selected with indoxacarb for 27 generations had a 25 623.17-fold resistance level. The highly indoxacarb resistant population showed very high cross-resistance to spinosad and very low cross-resistance to bifenthrin and chlorpyrifos. The mean estimated h2 of resistance to indoxacarb was only 0.04. The number of generations predicted for the development of ten-fold indoxacarb resistance at a constant h2  = 0.04 against selection intensities of 10% and 95% were 346 and 36 (slope = 3.18), 237 and 25 (slope = 2.18) and 128 and 13 (slope = 1.18). However, at a constant slope = 1.18 and selection intensities of 10% and 95%, the same increase in indoxacarb resistance occurred after 128 and 13 (h2  = 0.04), 103 and 11 (h2  = 0.05), and 86 and 9 (h2  = 0.06) generations. CONCLUSION: This study revealed that P. solenopsis has a very high resistance to indoxacarb as a result of laboratory selection. Thus, resistance to this insecticide in the field may also occur. Indoxacarb resistance can be minimized by rotating it with bifenthrin and/or chlorpyrifos due to its very low cross-resistance to these insecticides and by avoiding its rotation with Spinosad, which has very high cross-resistance. © 2019 Society of Chemical Industry.

Eugenol, a Plant Volatile, Synergizes the Effect of the Thrips Attractant, Ethyl Iso-Nicotinate.

The onion thrips, Thrips tabaci Lindeman (Thysanoptera: Thripidae), is a polyphagous pest that causes serious damage to agricultural crops, vegetables, and ornamental plants worldwide. Farmers rely on the extensive usage of synthetic chemical insecticides to control T. tabaci. There is a dire need to develop alternative control strategies to overcome the problems posed by chemical insecticides. Efficient traps would allow sensitive monitoring and possibly mass trapping. A field experiment was conducted to evaluate the potential of three plant compounds with known release rates (ranging from 6-30 mg/d); eugenol (Eug), 1, 8-cineole (eucalyptol), and linalool in all possible combinations with a thrips attractant, ethyl iso-nicotinate (EI). A combination of EI with Eug increased the effect of EI by attracting 100% more thrips (effect size, 1.95) as compared to the control of EI alone. Catches in remaining treatments were lower and or not significantly different from EI alone. The results from our study could be used to develop improved volatile blends to be used for monitoring traps. Our data suggests that these traps could be effective even at very low populations.

Spirotetramat Resistance Selected in the Phenacoccus solenopsis (Homoptera: Pseudococcidae): Cross-Resistance Patterns, Stability, and Fitness Costs Analysis.

The Phenacoccus solenopsis Tinsley (Homoptera: Pseudococcidae) is a major agricultural and horticultural pest of crops throughout the world. To develop a better resistance management strategy for P. solenopsis, we conducted a study on life history parameters of different populations of this pest, one selected with spirotetramat (Spiro-SEL), an unselected (UNSEL) population, and their reciprocal crosses. We also studied the cross-resistance and the stability of spirotetramat resistance. The Spiro-SEL of P. solenopsis exhibited a 328.69-fold resistance compared to the susceptible population (Lab-PK). The Spiro-SEL population also displayed a moderate level of cross-resistance to profenofos and bifenthrin and a high level of cross-resistance to abamectin. Resistance to spirotetramat in Spiro-SEL was unstable in the absence of selection. The study of life history parameters showed that there was a significant reduction in fitness parameters of Spiro-SEL population with a relative fitness value of 0.14. There was a significant decrease in survival rate, pupal weight, fecundity, egg hatching percentage, male and female generation time, intrinsic rate of population increase of males and females, biotic potential, and mean relative growth rate. It is concluded that selection with spirotetramat had marked effect on resistance development in P. solenopsis and upon removal of selection pressure spirotetramat resistance declined significantly, indicating unstable resistance. Development of resistance led to high fitness costs for the spirotetramat-selected population. Our study may provide the basic information on spirotetramat resistance and its mechanism to help develop the resistance management strategies.

Study of Synergism, Antagonism, and Resistance Mechanisms in Insecticide-Resistant Oxycarenus hyalinipennis (Hemiptera: Lygaeidae).

Oxycarenus hyalinipennis Costa (Hemiptera: Lygaeidae) is an economic and key pest of the Malvaceae family widely distributed in the world. Significant field resistance to cypermethrin (26.69-fold), chlorpyrifos (32.60-fold), methomyl (10.87), acetamiprid (20.63-fold), fipronil (5.84-fold), and spirotetramat (116.02-fold) has been reported. Cypermethrin combined with methomyl and spirotetramat, methomyl with spirotetramat, acetamiprid with spirotetramat, and fipronil with spirotetramat had synergistic effects (combination index, (CI) in a laboratory population of O. hyalinipennis named Lab-PK. Methomyl combined with acetamiprid and acetamiprid with fipronil had synergistic effects on O. hyalinipennis in a field in Multan named Field-POP. Cypermethrin combined with methomyl and spirotetramat; chlorpyrifos with methomyl, acetamiprid, and spirotetramat; methomyl with acetamiprid and spirotetramat; and fipronil with spirotetramat also had synergistic effects on O. hyalinipennis in Field-POP. Enzyme inhibitors piperonyl butoxide and S,S,S-tri-n-butyl phosphorotrithioate significantly increased the toxicity of chlorpyrifos, methomyl, acetamiprid, and spirotetramat to O. hyalinipennis in Field-POP, suggesting a monooxygenase- and esterase-based resistance mechanism. However, fipronil did not synergize with PBO and DEF. This study suggests that insecticide mixtures showing synergism must be determined for insecticide resistance management and other strategies such as rotations, mosaics, and cultural control should also be considered for the management of O. hyalinipennis.