A mixture of mesotrione and atrazine harms adults and larvae of the predatory wasp Polistes satan.

Herbicides have been intensively used for weed control, raising concerns about their potentially adverse effects on non-target organisms. Research on the effects of these common agrochemicals on beneficial insects and the ecosystem services they provide (e.g., predation and pollination) is scarce. Therefore, we tested whether a commercial formulation comprising a mixture of mesotrione and atrazine was detrimental to adult females and larvae of the Neotropical predatory social wasp Polistes satan, which is an effective natural enemy of crop pests. Wasps were individually fed syrups contaminated with different concentrations of the herbicide above and below the maximum label rate (MLR = 12 mL/L). Survival was assessed. The locomotor activity, immune response, and midgut morphology of adults as well as the immune response of the larvae were also studied. Herbicide concentrations far above the MLR (12, 40, and 100 times) caused adult mortality, whereas lower concentrations (0.5, 1, and 6 times) did not. Herbicide exposure at 0.5 to 12 times the MLR increased adult activity. Adult exposure at 0.1 or 0.5 times the MLR did not affect melanotic encapsulation of foreign bodies but led to changes in the morphology of the midgut epithelium and peritrophic matrix. In larvae, the ingestion of herbicide at 0.1 or 0.2 times the MLR (corresponding to 9.6 and 19.2 ng of herbicide per individual) did not cause mortality but decreased their melanization-encapsulation response. Increased locomotor activity in herbicide-exposed adults can affect their foraging activity. The altered midgut morphology of adults coupled with the decreased immune response in larvae caused by herbicide exposure at realistic concentrations can increase the susceptibility of wasps to infections. Therefore, herbicides are toxic to predatory wasps.

Effects of acephate and glyphosate-based agrochemicals on the survival and flight of Plebeia lucii Moure, 2004 (Apidae: Meliponini).

The conservation of terrestrial ecosystems depends largely on the preservation of pollinators, mainly bees. Stingless bees are among the main pollinators of native plants and crops in tropical regions, where they can be exposed to agrochemicals while foraging on contaminated flowers. In the present study, we investigated the effects on stingless bees of both a commonly used insecticide and herbicide in Brazil. Plebeia lucii Moure, 2004 (Apidae: Meliponini) foragers were orally chronically exposed to food contaminated with different concentrations of commercial formulations of the insecticide acephate or the herbicide glyphosate. Bee mortality increased with increasing agrochemical concentrations. Depending on its concentration, the acephate-based formulation reduced the lifespan and impaired the flight ability of bees. The glyphosate-based formulation was toxic only under unrealistic concentrations. Our results demonstrate that realistic concentrations of acephate-based insecticides harm the survival and alter the mobility of stingless bees. The ingestion of glyphosate-based herbicides was safe for forager bees under realistic concentrations.

The survival and flight capacity of commercial honeybees and endangered stingless bees are impaired by common agrochemicals.

The impact of agrochemicals on native Brazilian bees may be underestimated, since studies of non-target effects on bees have, by and large, concerned mostly the Apis mellifera L. Furthermore, bees may be exposed in the field to multiple agrochemicals through different routes, thus suggesting the necessity for more comprehensive toxicological experiments. Here, we assessed the lethal and sublethal toxicity of multiple agrochemicals (herbicide [glyphosate - Roundup®], fungicide [mancozeb], insecticide [thiamethoxam]) through distinct routes of exposure (contact or ingestion) to an endangered native Brazilian bee Melipona (Michmelia) capixaba Moure & Camargo, 1994 and to A. mellifera. Results indicate that none of the agrochemicals caused feeding repellency on the bees. Thiamethoxam caused high mortality of both species, regardless of the route of exposure or the dose used. In addition, thiametoxam altered the flight capacity of M. capixaba when exposed to the lowest dose via contact exposure. The field dose of glyphosate caused high mortality of both bee species after oral exposure as well as impaired the flight capacity of A. mellifera (ingestion exposure) and M. capixaba (contact exposure). The lower dose of glyphosate also impaired the flight of M. capixaba through either routes of exposure. Exposure of A. mellifera through contact and ingestion to both doses of mancozeb caused high mortality and significantly impaired flight capacity. Taken altogether, the results highlight the importance of testing the impact of multiple agrochemicals (i.e. not just insecticides) through different routes of exposure in order to understand more comprehensively the potential risks for Apis and non-Apis bees.

A predatory social wasp does not avoid nestmates contaminated with a fungal biopesticide.

Fungus-based biopesticides have been used worldwide for crop pest control as a safer alternative to chemical pesticides such as neonicotinoids. Both agrochemicals can be lethal and may also trigger side effects on the behavioral traits of non-target social insects, which play a crucial role in providing essential biological pest control services in agroecosystems. Here, we evaluated whether a commercial formulation of the entomopathogenic fungus Beauveria bassiana or the neonicotinoid imidacloprid causes mortality in foragers of Mischocyttarus metathoracicus. These social wasps are natural enemies of caterpillars and other herbivorous insects and inhabit both urban and agricultural environments in Brazil. We also tested whether wasps discriminate between biopesticide-exposed and unexposed conspecifics. Through a combination of laboratory (survival assay) and field experiments (lure presentation), along with chemical analyses (cuticular hydrocarbon profiles), we showed that topic exposure to the label rate of each pesticide causes a lethal effect, with the biopesticide exhibiting a slower effect. Moreover, wasps do not discriminate biopesticide-exposed from unexposed conspecifics, likely because of the similarity of their cuticular chemical profiles 24 h after exposure. Overall, the delayed lethal time at the individual level, combined with the indistinctive chemical cues of exposure and the lack of discrimination by conspecifics suggests that the fungal biopesticide may ultimately pose a threat to the colony survival of this predatory wasp.

Ingesting microplastics or nanometals during development harms the tropical pollinator Partamona helleri (Apinae: Meliponini).

The disposal of plastics and metal-derived compounds results in the contamination of the environment with nano/microparticles, leading to the exposure of various organisms to these harmful particles. However, the impacts of these particles on pollinating insects, which provide relevant ecosystem services, are not well understood. The aim of this study was to assess the effects of microscopic particles on the tropical pollinator Partamona helleri (Apinae: Meliponini), specifically evaluating the toxicity of plastic microparticles (polystyrene - PS, and polyethylene terephthalate - PET) and nanoparticles of a metal oxide (titanium dioxide - TiO2) via larval ingestion by bees reared in vitro. The survival rate of P. helleri larvae was not affected by the ingestion of particles of PS (500 ng/bee), PET (500 ng/bee), or TiO2 (10 µg/bee) compared to the non-treated diet (control or diet without the particles). Adults derived from treated larvae had increased body weight compared to the control, and the walking behavior of adults was altered by the ingestion of particles. Adults that ingested PET or TiO2 as larvae tended to rest for a longer time and interact more with other bees than the control. Hemocyte counts also changed, with a shift in the proportion of plasmatocytes and prohemocytes in treated individuals. Our findings suggest that even levels considered low for honey bees of exposure to plastic microparticles or metal nanoparticles can harm the health and behavior of stingless bees.

The impact of early-life exposure to three agrochemicals on survival, behavior, and gut microbiota of stingless bees (Partamona helleri).

Over the last few decades, agrochemicals have been partially associated with a global reduction in bees' population. Toxicological assessment is therefore crucial for understanding the overall agrochemical risks to stingless bees. Therefore, the lethal and sublethal effects of agrochemicals commonly used in crops (copper sulfate, glyphosate, and spinosad) on the behavior and gut microbiota of the stingless bee, Partamona helleri, were assessed using chronic exposure during the larval stage. When used at the field-recommended rates, both copper sulfate (200 µg of active ingredient/bee; a.i µg bee-1) and spinosad (8.16 a.i µg bee-1) caused a decrease in bee survival, while glyphosate (148 a.i µg bee-1) did not show any significant effects. No significant adverse effects on bee development were observed in any treatment with CuSO4 or glyphosate, but spinosad (0.08 or 0.03 a.i µg bee -1) increased the number of deformed bees and reduced their body mass. Agrochemicals changed the behavior of bees and composition of the gut microbiota of adult bees, and metals such as copper accumulated in the bees' bodies. The response of bees to agrochemicals depends on the class or dose of the ingested compound. In vitro rearing of stingless bees' larvae is a useful tool to elucidate the sublethal effects of agrochemicals.

Are Botanical Biopesticides Safe for Bees (Hymenoptera, Apoidea)?

The recent global decline in insect populations is of particular concern for pollinators. Wild and managed bees (Hymenoptera, Apoidea) are of primary environmental and economic importance because of their role in pollinating cultivated and wild plants, and synthetic pesticides are among the major factors contributing to their decline. Botanical biopesticides may be a viable alternative to synthetic pesticides in plant defence due to their high selectivity and short environmental persistence. In recent years, scientific progress has been made to improve the development and effectiveness of these products. However, knowledge regarding their adverse effects on the environment and non-target species is still scarce, especially when compared to that of synthetic products. Here, we summarize the studies concerning the toxicity of botanical biopesticides on the different groups of social and solitary bees. We highlight the lethal and sublethal effects of these products on bees, the lack of a uniform protocol to assess the risks of biopesticides on pollinators, and the scarcity of studies on specific groups of bees, such as the large and diverse group of solitary bees. Results show that botanical biopesticides cause lethal effects and a large number of sublethal effects on bees. However, the toxicity is limited when comparing the effects of these compounds with those of synthetic compounds.

Toxicological assessment of agrochemicals on bees using machine learning tools.

Machine learning (ML) is a branch of artificial intelligence (AI) that enables the analysis of complex multivariate data. ML has significant potential in risk assessments of non-target insects for modeling the multiple factors affecting insect health, including the adverse effects of agrochemicals. Here, the potential of ML for risk assessments of glyphosate (herbicide; formulation) and imidacloprid (insecticide, neonicotinoid; formulation) on the stingless bee Melipona quadrifasciata was explored. The collective behavior of forager bees was analyzed after in vitro exposure to agrochemicals. ML algorithms were applied to identify the agrochemicals that the bees have been exposed to based on multivariate behavioral features. Changes in the in situ detection of different proteins in the midgut were also studied. Imidacloprid exposure leads to the greatest changes in behavior. The ML algorithms achieved a higher accuracy (up to 91%) in identifying agrochemical contamination. The two agrochemicals altered the detection of cells positive for different proteins, which can be detrimental to midgut physiology. This study provides a holistic assessment of the sublethal effects of glyphosate and imidacloprid on a key pollinator. The procedures used here can be applied in future studies to monitor and predict multiple environmental factors affecting insect health in the field.

Ethoflow: Computer Vision and Artificial Intelligence-Based Software for Automatic Behavior Analysis.

Manual monitoring of animal behavior is time-consuming and prone to bias. An alternative to such limitations is using computational resources in behavioral assessments, such as tracking systems, to facilitate accurate and long-term evaluations. There is a demand for robust software that addresses analysis in heterogeneous environments (such as in field conditions) and evaluates multiple individuals in groups while maintaining their identities. The Ethoflow software was developed using computer vision and artificial intelligence (AI) tools to monitor various behavioral parameters automatically. An object detection algorithm based on instance segmentation was implemented, allowing behavior monitoring in the field under heterogeneous environments. Moreover, a convolutional neural network was implemented to assess complex behaviors expanding behavior analyses' possibilities. The heuristics used to generate training data for the AI models automatically are described, and the models trained with these datasets exhibited high accuracy in detecting individuals in heterogeneous environments and assessing complex behavior. Ethoflow was employed for kinematic assessments and to detect trophallaxis in social bees. The software was developed in desktop applications and had a graphical user interface. In the Ethoflow algorithm, the processing with AI is separate from the other modules, facilitating measurements on an ordinary computer and complex behavior assessing on machines with graphics processing units. Ethoflow is a useful support tool for applications in biology and related fields.

Spinosad-mediated effects in the post-embryonic development of Partamona helleri (Hymenoptera: Apidae: Meliponini).

The use of insecticides based on metabolites found in live organisms, such as the insecticide spinosad, has been an option for the control of agricultural pests because of the allegedly low toxicological risk for nontarget arthropods, such as stingless bees. In the current study, we evaluate the effects of chronic oral exposure to spinosad during the larval phase on survival, developmental time, body mass, midgut epithelial remodeling, and the peritrophic matrix (PM) of Partamona helleri stingless bee workers. Worker larvae that were raised in the laboratory were orally exposed to different concentrations (0, 6.53, 13.06, 32.64, and 3,264 ng. a.i. bee-1) of spinosad (formulation), and the resulting survival, developmental time, and body mass were studied. The concentration of spinosad recommended for use in the field (3,264 ng. a.i. bee-1) reduced the survival of workers during development. Also, sublethal concentrations of spinosad delayed the development and caused morphological changes in the midgut epithelium. Finally, the chronic exposure of larvae to 32.64 ng. a.i. bee-1 spinosad also altered the remodeling of the midgut during metamorphosis and affected the organization of the PM of larvae, pupae, and adults. Our data suggest possible environmental risks for using spinosad in cultures that are naturally pollinated by stingless bees.

Glyphosate is lethal and Cry toxins alter the development of the stingless bee Melipona quadrifasciata.

Brazil is the second largest producer of genetically modified plants in the world. This agricultural practice exposes native pollinators to contact and ingestion of Bacillus thuringiensis proteins (e.g. Cry toxins) from transgenic plants. Furthermore, native bees are also exposed to various herbicides applied to crops, including glyphosate. Little is known about the possible effects of glyphosate and Cry proteins on stingless bees, especially regarding exposure at an immature stage. Here, we show for the first time that glyphosate is lethal, and that Cry toxins (Cry1F, Cry2Aa) alter the development of the stingless bee Melipona quadrifasciata upon contamination of larval food. Glyphosate was very toxic to the bee larvae, killing all of them within only a few days of exposure. Bees treated with Cry2Aa proteins had a higher survival rate and were delayed in their development, compared to the negative controls. Those treated with the Cry1F protein also suffered delays in their development, compared to the negative controls. In conclusion, the proteins Cry1F, Cry2Aa, and the herbicide glyphosate were highly toxic to the stingless bee M. quadrifasciata, causing lethal or sublethal effects which can severely impair colony growth and viability, and reduce pollination ability.

The reduced-risk insecticide azadirachtin poses a toxicological hazard to stingless bee Partamona helleri (Friese, 1900) queens.

Large-scale pesticide application poses a major threat to bee biodiversity by causing a decline in bee populations that, in turn, compromises ecosystem maintenance and agricultural productivity. Biopesticides are considered an alternative to synthetic pesticides with a focus on reducing potential detrimental effects to beneficial organisms such as bees. The production of healthy queen stingless bees is essential for the survival and reproduction of hives, although it remains unknown whether biopesticides influence stingless bee reproduction. In the present study, we investigated the effects of the biopesticide azadirachtin on the survival, behavior, morphology, development, and reproduction of queens of the stingless bee Partamona helleri (Friese, 1900). The neonicotinoid imidacloprid was used as a toxic reference standard. Queens were orally exposed in vitro to a contaminated diet (containing azadirachtin and imidacloprid) during development. Azadirachtin resulted in reduced survival, similarly to imidacloprid, altered development time, caused deformations, and reduced the size of the queens' reproductive organs. All of these factors could potentially compromise colony survival. Results from the present study showed azadirachtin posed a toxicological hazard to P. helleri queens.