Enzymes mediating resistance to lambda-cyhalothrin in Eriopis connexa (Coleoptera: Coccinellidae).

Resistance to widely used insecticide, lambda-cyhalothrin, was recently reported in the predatory lady beetle Eriopis connexa (Germar) (Coleoptera: Coccinellidae). However, to understand whether metabolic mechanisms underlie such resistance, synergism bioassays and in vitro studies were carried out by using inhibitors and model substrates for enzymatic assays, respectively. The LD50s estimated for susceptible and resistant populations ηg of lambda-cyhalothrin/insect, and thus, a 22-fold difference in resistance ratio. Synergism ratios for the susceptible population with piperonyl butoxide (PBO), diethyl maleate (DEM), triphenyl phosphate (TPP), and S,S,S-tributylphosphorotrithioate (DEF) were respectively 33.8-, 0.24-, 0.35-, and 4.25-fold, while for the resistant population, they were 1463.0-, 0.79-, 0.85-, and 282.6-fold, respectively. The synergized resistance ratios were 0.50-, 2.00-, 6.75-, and 8.77-fold with PBO, DEF, DEM, and TPP, respectively, while resistance was virtually suppressed with DEF. The esterase exhibited 4.16-, 4.03-, and 5.38-fold greater activity towards formation of α-naphthol, ß-naphthol, and 4-nitrophenol in the resistant population of E. connexa than in the susceptible population. The activity of esterase depended on concentrations of DEF applied, either using α-naphthol or ß-naphthol, which completely inhibited the activity at 636 ηM. The PBO inhibited the ß-naphthol formation in approximately 50%, suggesting it as inhibitor of esterases. The activities of glutathione-S-transferase were similar and corresponded to 0.36-0.47 ηmol(-1) min(-1)µg of protein, for S and R populations, respectively. Similarly, the activities of cytochrome P450-dependent microsomal monooxygenases were 0.04 and 0.05 ηmol(-1) min(-1)µg of protein. The native gel indicated that the formation of ß-naphthol was completely inhibited by methyl-paraoxon, but only partially inhibited by eserine, TPP, and PBO. Although other studies with DEF and PBO have demonstrated strong inhibition of type B carboxylesterase associated with insecticide resistance, the results reported here do not rule out metabolism by cytochrome P450-dependent microsomal monooxygenases as a factor conferring E. connexa resistance to lambda-cyhalothrin and confirmed that PBO may also act by inhibiting esterases of insects.

Response of different populations of seven lady beetle species to lambda-cyhalothrin with record of resistance.

Simultaneous use of biological and chemical controls is a valued and historic goal of integrated pest management, but has rarely been achieved. One explanation for this failure may be the inadequate documentation of field populations of natural enemies for insecticide tolerance or resistance because natural enemies surviving insecticide application do not create problems like resistant pest species. Therefore, this study investigated 31 populations of lady beetles (Coleoptera: Coccinellidae) regarding their susceptibility to lambda-cyhalothrin, a pyrethroid insecticide that is widely used in cotton and other crops to control lepidopteran and coleopteran pests that are not targeted as prey by lady beetles. The study focused on seven coccinellid species common in cotton fields Coleomegilla maculata De Geer, Cycloneda sanguinea (L.), Eriopis connexa Germar, Harmonia axyridis (Pallas), Hippodamia convergens Guérin-Méneville, Olla v-nigrum (Mulsant), and Brumoides foudrasi (Mulsant) and one lady beetle species [Curinus coeruleus Mulsant] from a non-cotton ecosystem for comparisons. Dose-mortality curves were estimated after topical treatment of adult lady beetles with lambda-cyhalothrin. Statistically significant variations in lady beetle susceptibility were observed between species and between populations of a given species. Seven and eighteen populations of lady beetles exhibited greater values of LD50 and LD90, respectively, than the highest recommended field rate of lambda-cyhalothrin (20g a.i./hectare≈0.2g a.i./L) for cotton fields in Brazil. Furthermore, based on LD50 values, 29 out of 30 tested populations of lady beetles exhibited ratios of relative tolerance varying from 2- to 215-fold compared to the toxicity of lambda-cyhalothrin to the boll weevil, Anthonomus grandis Boh. (Coleoptera: Curculionidae). Four populations of E. connexa were 10.5-37.7 times more tolerant than the most susceptible population and thus were considered to be resistant to lambda-cyhalothrin, the first record of resistance for this species. These findings demonstrate that field selection for resistance to lambda-cyhalothrin in common lady beetles is occurring, opening up possibilities to effectively integrate biological control where the popular insecticide lambda-cyhalothrin is used.

Spermatogenesis, changes in reproductive structures, and time constraint associated with insemination in Podisus nigrispinus.

Males of the predatory stinkbug, Podisus nigrispinus (Dallas) (Hemiptera: Pentatomidae), accomplish long and multiple matings. We hypothesize that this behavior is due to time requirement for spermatozoa production and their transference to the females. Thus, this work investigated the effect of mating status of males and mating duration on spermatozoa transference to the females and the location of spermatozoa in the male reproductive tract during mating. On females, morphological alterations on female spermatheca and associated structures during a mating were investigated. Analyses of male reproductive tract showed presence of spermatozoa in the lumen of vas deferens was independent of mating status (ca. virgin, 0, 12 and 24 h after having a full mating), indicating continuous spermatogenesis which is supported by the absence of a seminal vesicle for spermatozoa storage. Female spermatheca had no changes associated with the duration of mating. However, females exhibited spermathecal elastic duct swelling by 30 min of mating duration. The success of males in filling the female spermatheca with spermatozoa depends on duration of mating. Thus, the results indicate that multiple mating is a requirement for reproductive success in the species by transference of spermatozoa and accessory substances stored in the female spermathecal duct. Likewise, the long mating is a male requirement to transfer materials in appropriate amount to the female but it is not dependent on spermatozoa alone.

Dual resistance to lambda-cyhalothrin and dicrotophos in Hippodamia convergens (Coleoptera: Coccinellidae).

Insecticide resistance is usually associated with pests, but may also evolve in natural enemies. In this study, adult beetles of three distinct North American populations of Hippodamia convergens Guérin-Méneville, and the progeny of reciprocal crosses between the resistant and most susceptible population, were treated topically with varying concentrations of lambda-cyhalothrin and dicrotophos. In addition, the LD50s of both insecticides were applied in combination to resistant individuals. The developmental and reproductive performance of each population was assessed in the absence of insecticide exposure to compare baseline fitness. California and Kansas populations were susceptible to both materials, whereas Georgia (GA) beetles exhibited a resistance ratio (RR50) of 158 to lambda-cyhalothrin and 530 to dicrotophos. Inheritance of lambda-cyhalothrin resistance was X-linked, whereas inheritance of dicrotophos resistance was autosomal. Mortality of resistant beetles treated with a mixture of LD50s of both materials was twice that of those treated with lambda-cyhalothrin alone, but not significantly different from those receiving dicrotophos alone. Life history parameters were largely similar among populations, except that Georgia beetles had higher egg fertility relative to susceptible populations. We conclude that the high levels of resistance to lambda-cyhalothrin and dicrotophos in Georgia beetles reflect heavy loads of these insecticides in local environments, most likely the large acreage under intensive cotton cultivation.

[Podisus nigrispinus (Dallas) (Hemiptera: Pentatomidae) requires long matings for successful reproduction]. / Podisus nigrispinus requer cópulas longas para o sucesso reprodutivo.

The occurrence of multiple and long matings seem to play an important role in the reproduction of the predatory stinkbugs such as Podisus nigrispinus (Dallas). However, the mechanisms underlying these behaviors remain unclear. In this study, the reproduction of P. nigrispinus was investigated as function of females' choice for their mating pair based on male body size and previous mating status, risk of predation and duration of mating. The female mating behavior was studied with female having multiple, partial or no partner choice based on male body size. Time to initiate a mating and its duration was observed after pairing males and females under risk of predation imposed by the presence of the predatory wasp Polistes versicolor Oliver. In addition, aiming to determine the reasons for long lasting matings, female had their mating interrupted at different intervals. Our data indicated that P. nigrispinus females do not select male partners as function of their body size and mating status. Duration of mating, mating partners' choice, and female fecundity and fertility were not influenced by the male size and risk of predation imposed by the predatory wasps. Mating interrupted after 30, 60, 120 and 240 min resulted in only 0, 3.1, 7.7 and 34% of egg hatching compared to 74.2% under uninterrupted mating (338 to 671 min long). Therefore, the reproductive success of P. nigrispinus females does not depend on male size and male mating status, but requires long-lasting mating as a condition for adequate spermatozoa transference to females instead.

Podisus nigrispinus requer cópulas longas para o sucesso reprodutivo / Podisus nigrispinus (Dallas) (Hemiptera: Pentatomidae) requires long matings for successful reproduction

The occurrence of multiple and long matings seem to play an important role in the reproduction of the predatory stinkbugs such as Podisus nigrispinus (Dallas). However, the mechanisms underlying these behaviors remain unclear. In this study, the reproduction of P. nigrispinus was investigated as function of females' choice for their mating pair based on male body size and previous mating status, risk of predation and duration of mating. The female mating behavior was studied with female having multiple, partial or no partner choice based on male body size. Time to initiate a mating and its duration was observed after pairing males and females under risk of predation imposed by the presence of the predatory wasp Polistes versicolor Oliver. In addition, aiming to determine the reasons for long lasting matings, female had their mating interrupted at different intervals. Our data indicated that P. nigrispinus females do not select male partners as function of their body size and mating status. Duration of mating, mating partners' choice, and female fecundity and fertility were not influenced by the male size and risk of predation imposed by the predatory wasps. Mating interrupted after 30, 60, 120 and 240 min resulted in only 0, 3.1, 7.7 and 34 percent of egg hatching compared to 74.2 percent under uninterrupted mating (338 to 671 min long). Therefore, the reproductive success of P. nigrispinus females does not depend on male size and male mating status, but requires long-lasting mating as a condition for adequate spermatozoa transference to females instead.

A realização de múltiplas e longas cópulas parece ser importante para a reprodução de percevejos predadores como Podisus nigrispinus (Dallas), mas o comportamento governando essas características permanece desconhecido. Neste estudo foram investigados o comportamento de cópula e a reprodução de P. nigrispinus em função do tamanho do macho, seu status prévio de acasalamento, risco de predação e interrupção de cópulas. A escolha da fêmea para o acasalamento foi estudada sob múltipla escolha, parcial ou sem chance de escolha por machos grandes ou pequenos. O comportamento para o início do acasalamento e tempo de cópula sob risco de predação quando na presença de Polistes versicolor Oliver foi comparado àquele de casais sem risco de predação. Machos e fêmeas de P. nigrispinus acasalaram independentemente do tamanho e status prévio de acasalamento dos machos, além de não apresentarem preferência por parceiros. A duração da cópula, escolha do parceiro e fertilidade das fêmeas não foram influenciadas pelo tamanho do macho, nem a presença do predador alterou o comportamento de acasalamento e duração de cópula. Fêmeas com cópulas interrompidas após 30, 60, 120 e 240 min apresentaram baixa viabilidade de ovos (0; 3,1; 7,7 e 34 por cento, respectivamente), enquanto aquelas com tempo natural de cópula (338 a 671 min) obtiveram 74,2 por cento de viabilidade. Portanto, o sucesso reprodutivo de P. nigrispinus não depende do tamanho do macho ou de seu status de acasalamento, mas requer longas cópulas para a transferência de espermatozóides.