Susceptibility monitoring and comparative gene expression of susceptible and resistant strains of Spodoptera frugiperda to lambda-cyhalothrin and chlorpyrifos.

BACKGROUND: Spodoptera frugiperda (J. E. Smith) is a widespread agricultural pest with several records of resistance to different insecticides and Bt proteins, including the neurotoxic insecticides chlorpyrifos (organophosphate) and lambda-cyhalothrin (pyrethroid). Here, we (i) characterized and monitored the susceptibility of field populations of S. frugiperda to chlorpyrifos (194 populations) and lambda-cyhalothrin (197 populations) collected from major maize-growing regions of Brazil from 2003 to 2016, and (ii) compared gene expression levels of laboratory-selected, chlorpyrifos- and lambda-cyhalothrin-resistant strains to a susceptible reference strain (Sf-ss) of S. frugiperda. RESULTS: The susceptibility monitoring detected average survival ranging from 29.3% to 36.0% for chlorpyrifos, and 23.1% to 68.0% for lambda-cyhalothrin. The resistance ratio of the chlorpyrifos-resistant strain (Clo-rr) was 25.4-fold and of the lambda-cyhalothrin-resistant strain (Lam-rr) was 21.5-fold. We identified 1098 differentially expressed genes (DEGs) between Clo-rr and Sf-ss, and 303 DEGs between Lam-rr and Sf-ss. Functional analyses of the DEGs revealed the up-regulation of several detoxification enzymes, mainly cytochrome P450 belonging to CYP3 and CYP6 clans. Genes associated with regulatory processes, such as the forkhead box class O (FoxO) transcription factor were also up-regulated. Variant analysis of target-site mutations for both pesticides identified the A201S and F290V mutations in acetylcholinesterase-1, both occurring in heterozigosis in the Clo-rr S. frugiperda strain. CONCLUSION: Our data show that the overexpression of the enzymatic detoxification machinery is the main difference to explain the resistance of Clo-rr and Lam-rr strains of S. frugiperda to chlorpyrifos and lambda-cyhalothrin, although a target-site mutation also contributes to the Clo-rr resistance to chlorpyrifos. © 2023 Society of Chemical Industry.

Fungivorous mites enhance the survivorship and development of stingless bees even when exposed to pesticides.

Stingless bees are the largest group of eusocial bees in the world. They play an essential role as crop pollinators and have been considered for inclusion in pesticide risk assessments (RAs). Beyond the mutualism involving stingless bee larvae and fungi, the fungivorous mite Proctotydaeus (Neotydeolus) alvearii proved to be interesting for studies of associations with stingless bees. Their presence is related to colony strength and health, showing a permanent-host-association level. Here, we tested whether the coexistence with P. (N.) alvearii affects stingless bee larvae survivorship and development, including when fed pesticide-dosed food. We chose dimethoate, the reference standard for toxicity tests, and thiamethoxam, widely used in neotropical crops and listed to be reassessed in RAs. Bees associated with the mites showed higher larval survivorship rates, even in the dosed ones, and revealed changes in the developmental time and body size. Our study represents the first approach to stingless bee responses to the coexistence of fungivorous mites inside brood cells, leading us to believe that these mites play a beneficial role in stingless bees, including when they are exposed to pesticides.

Laboratory Risk Assessment of Three Entomopathogenic Fungi Used for Pest Control toward Social Bee Pollinators.

The use of fungal-based biopesticides to reduce pest damage and protect crop quality is often considered a low-risk control strategy. Nevertheless, risk assessment of mycopesticides is still needed since pests and beneficial insects, such as pollinators, co-exist in the same agroecosystem where mass use of this strategy occurs. In this context, we evaluated the effect of five concentrations of three commercial entomopathogenic fungi, Beauveria bassiana, Metarhizium anisopliae, and Cordyceps fumosorosea, by direct contact and ingestion, on the tropical stingless bees Scaptotrigona depilis and Tetragonisca angustula, temperate bee species, the honey bee Apis mellifera, and the bumble bee Bombus terrestris, at the individual level. Furthermore, we studied the potential of two infection routes, either by direct contact or ingestion. In general, all three fungi caused considerable mortalities in the four bee species, which differed in their response to the different fungal species. Scaptotrigona depilis and B. terrestris were more susceptible to B. bassiana than the other fungi when exposed topically, and B. terrestris and A. mellifera were more susceptible to M. anisopliae when exposed orally. Interestingly, increased positive concentration responses were not observed for all fungal species and application methods. For example, B. terrestris mortalities were similar at the lowest and highest fungal concentrations for both exposure methods. This study demonstrates that under laboratory conditions, the three fungal species can potentially reduce the survival of social bees at the individual level. However, further colony and field studies are needed to elucidate the susceptibility of these fungi towards social bees to fully assess the ecological risks.

A food-ingested sublethal concentration of thiamethoxam has harmful effects on the stingless bee Melipona scutellaris.

In recent years, the importance of bee's biodiversity in the Neotropical region has been evidencing the relevance of including native bees in risk assessments. Therefore, the sublethal effects of the insecticide thiamethoxam on the survival and morphological parameters of the stingless bee Melipona scutellaris were investigated in the present study. Cells from both non-target organs (Malpighian tubules and midgut) and target organs (brain) were analyzed for morphological alterations using light microscopy and transmission electron microscopy. The findings showed that when M. scutellaris foragers were exposed to a sublethal concentration of thiamethoxam (LC50/100 = 0.000543 ng a. i./µL), longevity was not reduced but brain function was affected, even with the non-target organs attempting to detoxify. The cellular damage in all the organs was mostly reflected in irregular nuclei shape and condensed chromatin, indicating cell death. The most frequent impairments in the Malpighian tubules were loss of microvilli, disorganization of the basal labyrinth, and cytoplasmic loss. These characteristics are related to an attempt by the cells to increase the excretion process, probably because of the high number of toxic molecules that reach the Malpighian tubules and need to be secreted. In general, damages that compromise the absorption of nutrients, excretion, memory, and learning processes, which are essential for the survival of M. scutellaris, were found. The present results also fill in gaps on how these bees respond to thiamethoxam exposure and will be useful in future risk assessments for the conservation of bee biodiversity.

Ecological Modelling of Insect Movement in Cropping Systems.

The spatio-temporal dynamics of insect pests in agricultural landscapes involves the potential of species to move, invade, colonise, and establish in different areas. This study revised the dispersal of the important crop pests Diabrotica speciosa Germar and Spodoptera frugiperda (J.E. Smith) by using computational modelling to represent the movement of these polyphagous pests in agricultural mosaics. The findings raise significant questions regarding the dispersal of pests through crops and refuge areas, indicating that understanding pest movement is essential for developing strategies to predict critical infestation levels to assist in pest-management decisions. In addition, our modelling approach can be adapted for other insect species and other cropping systems despite discussing two specific species in the current manuscript. We present an overview of studies, combining experimentation and ecological modelling, discussing the methods used and the importance of studying insect movement as well as the implications for agricultural landscapes in Brazil.

Shifts in Ecological Dominance between Two Lepidopteran Species in Refuge Areas of Bt Cotton.

Competition behavior involving agricultural pest species has long been viewed as a powerful selective force that drives ecological and phenotypic diversity. In this context, a Game Theory-based approach may be useful to describe the decision-making dilemma of a competitor with impacts to guarantee its superiority in terms of ecological dominance or sharing of the food resource with its competitor. In an attempt to elucidate the consequences of competitive dynamics for the ecological dominance of these species in refuge areas of Bt cotton, we conducted a study that was divided into two parts. The first study consisted of an evaluation of interactions involving Spodoptera frugiperda (JE Smith, 1797) and Helicoverpa armigera (Hübner, 1808) on non-Bt cotton plants in a field trial. In the second study, we explored the data matrix collected in the field to parameterize a model of Cellular Automata (CA) with update rules inspired by Game Theory. Computer simulations were analyzed in hypothetical scenarios involving the application (or not) of insecticides in the refuge areas in combination with the resistance factor of one or both pest species to the insecticides used in the refuge areas. H. armigera had superior competitive performance in relation to S. frugiperda only at high densities. According to the density-mediated shift in dominance of the species, the resistance of S. frugiperda to insecticides is seen as a risk factor for the production of susceptible individuals of H. armigera on a large scale in the refuge areas. Additionally, S. frugiperda insecticide resistance may potentially impact the resistance evolution of the H. armigera population to Bt cotton. Thus, ecological dominance could diverge by the presence of a resistance allele to insecticides with interspecific competition perhaps subordinate to evolutionary processes.

Apis mellifera and Melipona scutellaris exhibit differential sensitivity to thiamethoxam.

Apis mellifera is a pollinator insect model in pesticide risk assessment tests for bees. However, given the economic and ecological importance of stingless bees such as Melipona scutellaris in the Neotropical region, as well as the lack of studies on the effect of insecticides on these bees, toxicity tests for stingless bees should be carried out to understand whether insecticides affect both species of bees in the same manner. Thus, the present study quantified the differential sensitivity of the bees M. scutellaris and A. mellifera to the oral ingestion of the insecticide thiamethoxam by determining the mean lethal concentration (LC50), mean lethal time (LT50), and their effect on the insecticide target organ, the brain. The results showed that the stingless bee is more sensitive to the insecticide than A. mellifera, with a lower LC50 of 0.0543 ng active ingredient (a.i.)/µL for the stingless bee compared to 0.227 ng a.i./µL for A. mellifera. When exposed to a sublethal concentration, morphological and ultrastructural analyses were performed and evidenced a significant increase in spaces between nerve cells of both species. Thus, A. mellifera is not the most appropriate or unique model to determine the toxicity of insecticides to stingless bees.

The impact of four widely used neonicotinoid insecticides on Tetragonisca angustula (Latreille) (Hymenoptera: Apidae).

Application of neonicotinoid insecticides on crops can reduce the pollination services and population levels of the stingless bee Tetragonisca angustula (Latreille) (Hymenoptera: Apidae) in Neotropical agroecosystems. However, the impact of these insecticides on this bee has not been fully investigated. This study assessed the susceptibility levels of T. angustula to four neonicotinoid insecticides (acetamiprid, imidacloprid, thiacloprid and thiamethoxam), widely used to manage pests on different crops, and their effects on locomotion of the bee. Neonicotinoids with the cyano radical caused lower bee mortality (assessed by mean lethal concentration, LC50), while those compounds with the nitro radical were highly harmful to T. angustula. Locomotion activity was strongly impaired in bees treated with thiacloprid and imidacloprid, while acetamiprid did not affect the locomotion activity, which was similar to the control. Application of thiamethoxam caused hyperactivity, as observed by increases of ∼4.5 and 5.0-fold in mean speed and distance traveled, respectively. These results suggest that applications of neonicotinoid insecticides can negatively affect the pollination activity of T. angustula. These results can also help to develop strategies to conserve these pollinators in agroecosystems.

Late effect of larval co-exposure to the insecticide clothianidin and fungicide pyraclostrobin in Africanized Apis mellifera.

Among the factors that contribute to the reduction of honeybee populations are the pesticides. These chemical compounds reach the hive through forager bees, and once there, they can be ingested by the larvae. We evaluated the effects of repeated larval exposure to neonicotinoid insecticide, both in isolation and in combination with strobilurin fungicide, at environmentally relevant doses. The total consumption of the contaminated diet was 23.63 ng fungicide/larvae (pyraclostrobin) and 0.2364 ng insecticide/larvae (clothianidin). The effects on post-embryonic development were evaluated over time. Additionally, we assessed the survival pattern of worker bees after emergence, and the pesticides' effects on the behavior of newly emerged workers and young workers. Young bees that were exposed to the fungicide and those subjected to co-exposure to both pesticides during larval phase showed behavioral changes. The insecticide, both in isolation and in combination with fungicide reduced the bees' longevity; this effect of larval exposure to pesticides was stronger in bees that were exposed only to the insecticide. Although the larvae did not have sensitivity to exposure to pesticides, they showed later effects after emergence, which may compromise the dynamics of the colony, contributing to the reduction of the populations of bees in agroecosystems.

Food extraction by the males of Podisus nigrispinus (Dallas) (Hemiptera: Pentatomidae) from cotton leafworm larvae

In this work, the effect of different densities (1, 3, 5, 7 and 9) of 3rd instar Alabama argillacea (Huebner) larvae on food consumption by Podisus nigrispinus (Dallas) males was evaluated. The densities established were converted to weight of prey offered: 13.4 mg (one larva), 33.3 mg (three larvae), 54.3 mg (five larvae), 81.8 mg (seven larvae), and 110.34 mg (nine larvae). The quantity of food consumed by P. nigrispinus increased with the prey density. The density of preys did not affect the time spent by the predator to ingest the food. The quantity of food extracted per minute was always higher in smaller densities and lower in higher densities. Males mean body weight did not differ statistically between the treatments tested, and weight gain was smaller in the first two densities tested. Relative consumption rates increased with the quantity of larvae offered. P. nigrispinus males might change its predatory behavior as a function of the quantity of prey available.

O efeito de diferentes densidades (1, 3, 5, 7 e 9) de 3º lagartas do instar de Alabama argillacea (Huebner) no consumo de alimento por machos de Podisus nigrispinus (Dallas) foi avaliado em condições de laboratório. As densidades utilizadas foram convertidas a peso de presa oferecida: 13,40 mg (1 lagarta), 33.30 mg (3 lagartas), 54,30 mg (5 lagartas), 81,80 mg (7 lagartas) e 110,34 mg (9 lagartas). A quantidade de alimento consumido pelo macho de P. nigrispinus aumentou com a densidade da presa. A densidade da presa não afetou o tempo gasto pelo predador para ingerir o alimento. A quantidade de alimento extraído por minuto pelo predador foi sempre maior nas mais baixas densidades e menores nas mais altas densidades. O peso corpóreo médio dos machos de P. nigrispinus não difereriu entre os tratamentos testados e o seu ganho de peso foi menor nas duas primeiras densidades testadas. As taxas de consumo relativo aumentaram com a quantidade de lagartas oferecidas. Os machos de P. nigrispinus podem mudar seu comportamento predatório em função da quantidade de presas disponíveis.

Age-dependent fecundity of Podisus nigrispinus (Dallas) (Heteroptera: Pentatomidae) with sublethal doses of gammacyhalothrin

Podisus nigrispinus (Dallas) (Heteroptera: Pentatomidae) was exposed to gammacyhalothrin to study the stimulatory effect of low concentrations of this pyrethroid. The doses studied were 23.4375, 11.7188, 5.8594, 2.9297, 1.4648, 0.7324 mg.litre-1 and water, as control. The third, fourth, and fifth instars of P. nigrispinus were shorter with all the doses of this insecticide. The survival and longevity of this predator was highest with the lowest doses. The oviposition period of P. nigrispinus decreased as the doses increased. The lowest pre-and postoviposition periods were found with 11.7188 mg.litre-1 of gammacyhalothrin. The dose 0.7324 mg.litre-1 increased the clutch interval. The egg viability was similar between the treatments. The number of eggs per clutch, clutches per female, incubation period, and female body weight were not affected. The hormesis response could be used as a tool for IPM programs, allowing the manipulating parameters of this predator in synergism with this insecticide.

Estudou-se o efeito estimulatório de baixas concentrações de gamacihalotrin sobre Podisus nigrispinus (Dallas)(Heteroptera: Pentatomidae). As doses de gamacihalotrin estudadas foram: 23,4375, 11,7188, 5,8594, 2,9297, 1,4648, 0,7324 mg.litro-1 e água (controle). Todas as doses estudadas de gamacihalotrin reduziram o período de desenvolvimento do 3º, 4º e 5º ínstares de P. nigrispinus. A sobrevivência e a longevidade do predador foram mais altas quando foi exposto a baixas doses. O período de oviposição de P. nigrispinus reduziu á medida que foram aumentadas as doses de gamacihalotrin. A dose de 11,7188 mg.litro-1 de gamacihalotrin causou os mais curtos períodos de pré-oviposição e pós-oviposição. O intervalo entre posturas aumentou com a dose de 0,7324 mg.litro-1 de gamacihalotrin. A viabilidade de ovos não foi afetada pelo piretróide. O número de ovos por postura, posturas por fêmea, período de incubação e peso corpóreo de fêmea não foram afetados pelas doses testadas de gamacyhalotrin.