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.

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.

Investigating the effects of mesotrione/atrazine-based herbicide on honey bee foragers.

A mixture of the herbicides mesotrione and atrazine (Calaris®) is a widely used herbicide in agriculture in several countries. However, the possible toxicological effects of this formulation on non-target organisms require investigation. In this study, the effects of acute oral exposure to Calaris® were evaluated in Apis mellifera foragers. The effect of seven different concentrations of Calaris® on survival and sucrose consumption was studied, while the recommended concentration for field use (FC) and its 10× dilution (0.1 FC) were used to assess overall locomotor activity, respiratory rate, flight, midgut morphology, oxidative and nitrosative stresses, and hemocyte counting. The exposure to FC or 0.1 FC decreased locomotor activity and induced damage to the midgut epithelium. Additionally, the two tested concentrations reduced superoxide dismutase activity, nitric oxide levels, and total hemocyte count. FC also increased malondialdehyde content and 0.1 FC increased respiratory rate and decreased the proportion of prohemocytes. Overall, our findings evidenced significant harmful effects on A. mellifera foragers resulting from the ingestion of the Calaris® herbicide.

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.

Ingestion of polystyrene microparticles impairs survival and defecation in larvae of Polistes satan (Hymenoptera: Vespidae).

Microplastics (MPs) are widespread pollutants of emerging concern, and the risks associated with their ingestion have been reported in many organisms. Terrestrial environments can be contaminated with MPs, and terrestrial organisms, including arthropods, are predisposed to the risk of ingesting MPs. In the current study, the larvae of the paper wasp Polistes satan were fed two different doses (6 mg or 16 mg at once) of polystyrene MPs (1.43 mm maximum length), and the effects of these treatments on immature development and survival till adult emergence were studied. Ingestion of the two doses resulted in mortality due to impaired defecation prior to pupation. The survival of larvae that ingested 16 mg of MPs was significantly lower than that of the control. The ingestion of 16 mg of MPs also reduced the adult emergence (11.4%) in comparison to the control (44.4%). MPs were not transferred from the larvae to the adults that survived. These findings demonstrate that MP ingestion can be detrimental to P. satan, e.g. larval mortality can decrease colony productivity and thus the worker force, and that MPs can potentially affect natural enemies that occur in crops, such as predatory social wasps.

Fipronil exposure compromises respiration and damages the Malpighian tubules of the stingless bee Partamona helleri Friese (Hymenoptera: Apidae).

Fipronil has been widely used in agriculture and forestry in Brazil to control several pests. However, this insecticide may be hazardous to non-target organisms, including stingless bees, which are essential pollinators of crops and natural environments. Here, we investigated the effect of 24-h acute oral exposure to LC50 of fipronil on the Malpighian tubules of the stingless bee Partamona helleri (Friese). Insecticide exposure decreases the respiration rate of forager bees, and the Malpighian tubules are severely affected, as shown by the epithelial architecture disorganization, loss of cytoplasmic content, degradation of the brush border, and nuclear pyknosis. In addition, fipronil ingestion increases the number of Malpighian cells positive for peroxidase, LC3, cleaved caspase-3, and JNK. However, Notch and ERK1/2-positive cells decrease in the exposed bees. These changes in the signaling proteins indicate an increase in oxidative stress, autophagy and apoptosis, and impairment of cell recovery. Overall, our results demonstrate the toxicological effects of fipronil on a stingless bee, which compromises the physiology of this important pollinator.

Heat stress negatively affects physiology and morphology during germination of Ormosia coarctata (Fabaceae, Papilionoideae).

Research on the morphophysiological behavior of forest seeds during germination with respect to climate change is scarce. To date, there have been no studies on the biochemical or morphological aspects of Ormosia spp. In this study, we subjected Ormosia coarctata seeds to various temperature conditions to investigate temperature-dependent impacts on morphology, reactive oxygen species (ROS) generation, antioxidant systems, and storage systems. Analyses were performed on seeds exposed to 25, 35, and 40 °C for 48, 96, and 144 h. The morphology was evaluated by radiation using a Faxitron MX-20 device. ROS production (superoxide anion and hydrogen peroxide), malonaldehyde (MDA), carbonylated proteins, antioxidant enzyme activity (superoxide dismutase [SOD], ascorbate peroxidase [APX], catalase [CAT], and peroxidase [POX]), ß-carotene, lycopene, glucose, and reserve enzyme activity (α- and ß-amylase, lipase, and protease) were analyzed by spectrophotometry. Heat stress (40 °C) decreased germination by 76.2% and 78.1% (compared to 25 and 35 °C, respectively), caused damage to the external morphology of the seed, increased the content of ROS, MDA, and carbonylated proteins, and reduced APX, CAT, and POX activity. Furthermore, heat stress decreased glucose content and α-amylase activity. These results suggest that an increase of 5 °C in temperature negatively affects germination, promotes oxidative stress, and induces deterioration in O. coarctata seeds.

Exposure to copper sulfate impairs survival, post-embryonic midgut development and reproduction in Aedes aegypti.

Aedes aegypti is a vector of several global human viruses responsible for high human morbidity and mortality. The method to prevent the transmission of vector-borne viruses is mainly based on the control of the insect vector using insecticides. Among these chemicals, copper sulfate is a compound widely used in agriculture with the potential to be used as an alternative to control these insects. This study evaluated the effects of the exposure of A. aegypti larvae to copper sulfate on survival, midgut morphology, blood-feeding and fecundity. The exposure to CuSO4 decreased the survival of A. aegypti during the immature phase. Adults obtained from exposed larvae had their lifespan decreased at all tested concentrations. The exposure to CuSO4 impaired the development in the transition from larvae to pupae and from pupae to adult. The number of eggs laid by females developed from larvae treated with CuSO4 was significantly lower than in control. In addition, the egg hatching rates were also negatively affected. The midguts of treated larvae and pupae showed epithelial disorganization. The number of cleaved caspase-3 cells increased in the midgut of exposed pupae compared to control. Moreover, there was a reduction in proliferating cells in treated larvae and pupae compared to the control. In conclusion, the results reveal that CuSO4 exposure has insecticidal activity against A. aegypti, which may be related to the impairment of the midgut metamorphosis and reduced proliferation of stem cells, with the consequent impairment of female mosquito fertility and fecundity.

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.

Impact of copper sulfate on survival, behavior, midgut morphology, and antioxidant activity of Partamona helleri (Apidae: Meliponini).

Copper sulfate (CuSO4) is widely used in agriculture as a pesticide and foliar fertilizer. However, the possible environmental risks associated with CuSO4 use, particularly related to pollinating insects, have been poorly studied. In this study, we evaluated both lethal and sublethal effects of CuSO4 on the stingless bee Partamona helleri. Foragers were orally exposed to five concentrations of CuSO4 (5000, 1666.7, 554.2, 183.4, 58.4 µg mL-1), and the concentration killing 50% (LC50) was estimated. This concentration (142.95 µg mL-1) was subsequently used in behavioral, midgut morphology, and antioxidant activity analyses. Bee mortality increased with the ingestion of increasing concentrations of CuSO4. Ingestion at the estimated LC50 resulted in altered walking behavior and damage to the midgut epithelium and peritrophic matrix of bees. Furthermore, the LC50 increased the catalase or superoxide dismutase activities and levels of the lipid peroxidation biomarker malondialdehyde. Furthermore, the in situ detection of caspase-3 and LC3, proteins related to apoptosis and autophagy, respectively, revealed that these processes are intensified in the midgut of treated bees. These data show that the ingestion of CuSO4 can have considerable sublethal effects on the walking behavior and midgut of stingless bees, and therefore could pose potential risks to pollinators including native bees. Graphical abstract.

A Genetically Modified Anti-Plasmodium Bacterium Is Harmless to the Foragers of the Stingless Bee Partamona helleri.

Paratransgenesis consists of genetically engineering an insect symbiont to control vector-borne diseases. Biosafety assessments are a prerequisite for the use of genetically modified organisms (GMOs). Assessments rely on the measurement of the possible impacts of GMOs on different organisms, including beneficial organisms, such as pollinators. The bacterium Serratia AS1 has been genetically modified to express anti-Plasmodium effector proteins and does not impose a fitness cost on mosquitoes that carry it. In the present study, we assessed the impact of this bacterium on the native bee Partamona helleri (Meliponini), an ecologically important species in Brazil. Serratia eGFP AS1 (recombinant strain) or a wild strain of Serratia marcescens were suspended in a sucrose solution and fed to foragers, followed by measurements of survival, feeding rate, and behavior (walking and flying). These bacteria did not change any of the variables measured at 24, 72, and 144 h after the onset of the experiment. Recombinant and wild bacteria were detected in the homogenates of digestive tract during the 144 h period analyzed, but their numbers decreased with time. The recombinant strain was detected in the midgut at 24 h and in the hindgut at 72 h and 144 h after the onset of the experiment under the fluorescent microscope. As reported for mosquitoes, Serratia eGFP AS1 did not compromise the foragers of P. helleri, an ecologically relevant bee.

Harmful effects of fipronil exposure on the behavior and brain of the stingless bee Partamona helleri Friese (Hymenoptera: Meliponini).

Fipronil is a pesticide widely used to control agricultural and household insect pests. However, fipronil is highly toxic to non-target insects, including pollinators. In this study, we investigated the acute effects of fipronil on the behavior, brain morphology, antioxidant activity, and proteins related to signaling pathways on the brain of workers of the stingless bee Partamona helleri. The ingestion of fipronil increases both the walking distance and velocity and causes enlarged intercellular spaces in the Kenyon cells and intense vacuolization in the neuropils of the brain. Moreover, fipronil decreases the activity of catalase (CAT) and increases the activity of glutathione S-transferase (GST). However, there is no difference in superoxide dismutase (SOD) activity between the control and fipronil. Regarding immunofluorescence analysis, bees exposed to fipronil showed an increase in the number of cells positive for cleaved caspase-3 and peroxidase, but a reduction in the number of cells positive for ERK 1/2, JNK and Notch, suggesting neuron death and impaired brain function. Our results demonstrate that fipronil has harmful effects on the behavior and brain of a stingless bee, which may threaten the individuals and colonies of this pollinator.

Heat stress-mediated effects on the morphophysiological, biochemical, and ultrastructural parameters of germinating Melanoxylon brauna Schott. seeds.

KEY MESSAGE: The present study showed that the heat stress (40 °C) caused changes in morphophysiological, biochemical, and ultrastructural parameters to the seeds Melanoxylon brauna, ultimately leading to loss of germination capacity. Temperature is an abiotic factor that influences seed germination. In the present study, we investigated morphophysiological, biochemical, and ultrastructural changes during the germination of Melanoxylon brauna seeds under heat stress. Seed germination was evaluated at constant temperatures of 25 and 40 °C. The samples consisted of seeds soaked in distilled and ionized water for 48 and 96 h at both temperatures. For the evaluation of internal morphology, the seeds were radiographed. Ultrastructural parameters were assessed using transmission electron microscopy (TEM). The production of reactive oxygen species (ROS), content of malondialdehyde (MDA) and glucose, carbonylated proteins, and activity of the enzymes (superoxide dismutase-SOD, ascorbate peroxidase-APX, catalase-CAT, peroxidase-POX, glucose-6-phosphate dehydrogenase-G6PDH, lipase, α- and ß-amylase, and protease) were measured by spectrophotometric analysis. An 82% reduction in the germination of M. brauna seeds was observed at 25 °C, and 0% at 40 °C. TEM showed that seeds submitted to heat stress (40 °C) had poorly developed mitochondria and significantly reduced respiration rates. The content of ROS and protein carbonylation in seeds subjected to 40 °C increased compared to that at 25 °C. The activity of antioxidant enzymes, namely SOD, APX, CAT, and POX, was significantly reduced in seeds subjected to heat stress. Glucose content, G6PDH, and lipase activity also decreased when the seeds were exposed to heat stress. Conversely, α- and ß-amylase enzymes and the protease increased due to the increase in temperature. Our data showed that the increase in temperature caused an accumulation of ROS, increasing the oxidative damage to the seeds, which led to mitochondrial dysfunction, ultimately leading to loss of germination.

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.

Effects of sub-lethal doses of nisin on the virulence of Salmonella enterica in Galleria mellonella larvae.

Salmonella enterica is a pathogen that induces self-limiting gastroenteritis and is of worldwide concern. Nisin, an antimicrobial peptide, has emerged as an alternative for the control of microbial growth but its effect on the virulence of pathogenic bacteria is not yet well-explored. This work aimed to evaluate the virulence of S. enterica in the presence of sub-inhibitory nisin using the experimental model Galleria mellonella. Sub-inhibitory concentrations of nisin of 11.72 and 46.88 µM did not affect the cellular viability of S. enterica but promoted changes in gene expression within 1 h of treatment, with increases of up to 3-fold of pagC, 1.8-fold of invA and 2.3-fold of invF. Larvae of G. mellonella inoculated with S. enterica combined with nisin at 46.88 µM presented mortality, and TL50 noticeably increased to 50% and 80% at 24 and 48 h post-infection, respectively. Defence responses, such as melanisation, nodulation, pseudopodia, immune response, and expression of defence proteins of the larvae G. mellonella were enhanced when the treatments with S. enterica were combined with 11.72 or 46.88 µM nisin. These results show an increase in virulence of S. enterica by sub-MIC concentration of nisin that needs to be explored.

Acute exposure to fipronil induces oxidative stress, apoptosis and impairs epithelial homeostasis in the midgut of the stingless bee Partamona helleri Friese (Hymenoptera: Apidae).

Partamona helleri is an important pollinator in natural and agricultural ecosystems in the neotropics. However, the foraging activity of this bee increases its risk of exposure to pesticides, which may affect both the individuals and the colony. Thus, this study aims to evaluate the side effects of LC50 of fipronil (0.28 ng a.i. µL-1) on the midgut morphology, antioxidant activity and some pathways of cell death, proliferation and differentiation in workers of P. helleri, after 24 h of oral exposure. Fipronil caused morphological alterations in the midgut of the bees. The activities of the detoxification enzymes superoxide dismutase, catalase and glutathione S-transferase increased after exposure, which suggests the occurrence of a detoxification mechanism. Furthermore, exposure to fipronil changed the number of positive cells for signaling-pathway proteins in the midgut of bees, which indicates the induction of cell death by the apoptotic pathway and impairment of the midgut epithelial regeneration. These results demonstrate that fipronil may negatively affect the morphology and physiology of the midgut of the stingless bee P. helleri and impose a threat to the survival of non-target organisms.

Biological, histological and immunohistochemical studies on the toxicity of spent coffee grounds and caffeine on the larvae of Aedes aegypti (Diptera: Culicidae).

The mosquito Aedes aegypti is a primary vector for major arboviruses, and its control is mainly based on the use of insecticides. Caffeine and spent coffee grounds (CG) are potential agents in controlling Ae. aegypti by reducing survival and blocking larval development. In this study, we analyzed the effects of treatment with common CG (CCG: with caffeine), decaffeinated CG (DCG: with low caffeine), and pure caffeine on the survival, behavior, and morphology of the midgut of Ae. aegypti under laboratory conditions. Third instar larvae (L3) were exposed to different concentrations of CCG, DCG, and caffeine. All compounds significantly affected larval survival, and sublethal concentrations reduced larval locomotor activity, delayed development, and reduced adult life span. Damage to the midgut of treated larvae included changes in epithelial morphology, increased number of peroxidase-positive cells (more abundant in DCG-treated larvae), and caspase 3-positive cells (more abundant in CCG-treated larvae), suggesting that the treatments triggered cell damage, leading to activation of cell death. In addition, the treatments reduced the FMRFamide-positive enteroendocrine cells and dividing cells compared to the control. CG and caffeine have larvicidal effects on Ae. aegypti that warrant field testing for their potential to control mosquitoes.

A mixture containing the herbicides Mesotrione and Atrazine imposes toxicological risks on workers of Partamona helleri.

A mixture of Mesotrione and Atrazine (Calaris®) has been reported as an improvement of the atrazine herbicides, which are agrochemicals used for weed control. However, its possible harmful effects on non-target organisms, including pollinators, needs to be better understood. In this work, the effects of the mix of herbicides on food consumption, behaviour (walking distance, and meandering), and the morphology of the midgut of the stingless bee Partamona helleri were studied. Foragers were orally exposed to different concentrations of the mix. The concentrations leading to 10% and 50% mortality (LC10 and LC50, respectively) were estimated and used in the analysis of behaviour and morphology. The ingestion of contaminated diets (50% aqueous sucrose solution + mix) led to a reduction in food consumption by the bees when compared to the control, bees fed a non-contaminated diet (sucrose solution). Ingestion of the LC50 diet reduced locomotor activity, increased meandering, induced the degradation of the epithelium and peritrophic matrix, and also changed the number of cells positive for signalling-pathway proteins in the midgut. These results show the potential toxicological effects and environmental impacts of the mix of herbicides in beneficial insects, including a native bee.