Bees are considered as important providers of ecosystem services, acting via pollination process in crops and native plants, and contributing significantly to the maintenance of biodiversity. However, the decrease of bee's population has been observed worldwide and besides other factors, this collapse is also related to the extensive use of pesticides. In this sense, studies involving the assessment of adverse effects and the uptake of pesticides by bees are of great concern. This work presents an analytical method for the determination of the insecticide abamectin and the fungicide difenoconazole in the stingless bee Melipona scutellaris exposed via oral and topic to endpoints concentrations of active ingredients (a.i.) alone and in commercial formulations and the discussion about its mortality and uptake. For this purpose, QuEChERS (Quick, Easy, Cheap, Efficient, Rugged and Safe) acetate modified method was used for extraction and pesticides were determined by LC-MS/MS. The validation parameters have included: a linear range between 0.01 and 1.00 μg mL⁻¹; and LOD and LOQ of 0.038 and 0.076 μg g⁻¹ for abamectin and difenoconazole, respectively. The uptake of tested pesticides via oral and topic was verified by the accumulation in adult forager bees, mainly when the commercial product was tested. Mortality was observed to be higher in oral exposure than in topic tests for both pesticides. For abamectin in a commercial formulation (a.i.) no differences were observed for oral or topic exposure. On the other hand, for difenoconazole, topic exposure had demonstrated higher accumulation in bees, according to the increase of received dose. Through the results, uptake and the possible consequences of bioaccumulated pesticides are also discussed and can contribute to the knowledge about the risks involving the exposure of bees to these compounds.

Pollinating bees are stressed by highly variable environmental conditions, malnutrition, parasites and pathogens, but may also by getting in contact with microorganisms or entomopathogenic nematodes that are used to control plant pests and diseases. While foraging for water, food, or nest material social as well as solitary bees have direct contact or even consume the plant protection product with its active substance (e.g., viruses, bacteria, fungi, etc.). Here, we summarize the results of cage, microcolony, observation hive assays, semi-field and field studies using full-size queen-right colonies. By now, some species and subspecies of the Western and Eastern honey bee (Apis mellifera, A. cerana), few species of bumble bees, very few stingless bee species and only a single species of leafcutter bees have been studied as non-target host organisms. Survival and reproduction are the major criteria that have been evaluated. Especially sublethal effects on the bees' physiology, immune response and metabolisms will be targets of future investigations. By studying infectivity and pathogenic mechanisms, individual strains of the microorganism and impact on different bee species are future challenges, especially under field conditions. Overall, it became evident that honey bees, bumble bees and few stingless bee species may not be suitable surrogate species to make general conclusions for biological mechanisms of bee-microorganism interactions of other social bee species. Solitary bees have been studied on leafcutter bees (Megachile rotundata) only, which shows that this huge group of bees (∼20,000 species worldwide) is right at the beginning to get an insight into the interaction of wild pollinators and microbial plant protection organisms.

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 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.

Currently, Brazil has a full framework for pesticide risk assessment established for Apis mellifera, based on the North American approach. However, the use of Apis mellifera as model-organism as a surrogate for Brazilian native species of stingless bees has been questioned. Assessments on other stages of development than adult individual are essential. Our study aimed to standardize in vitro larval rearing method for the stingless bee species Scaptotrigona postica and Tetragonisca angustula, comparing the results to those obtained for M. scutellaris (previously described), for proposing the most suitable one for using in toxicological larval tests. We used the most efficient method for determining the toxicity of dimethoate on S. postica larvae. We presented the first comparative approach of responses to in vitro larval rearing methods among native bee species from Neotropical region, for use in risk assessment. Our results showed that S. postica was the most suitable native species to be proposed as model-organism. In addition, our results are also very useful for a ring test to validate the method, in accordance to OECD.