Residual determination and acute toxicity of the neonicotinoid clothianidin in the neotropical stingless bee Tetragonisca angustula Latreille, 1811 (Apidae: Meliponini).

Bees play a crucial role as natural pollinators, ensuring the maintenance and stability of the world's biodiversity and agricultural crops. Native bees in neotropical regions belong to the Meliponini tribe, a larger group that differs significantly in behavior and biology from honeybees (e.g., Apis mellifera) and solitary bees (e.g., Osmia spp.). Hence, the exposure and effects of pesticides is also likely to vary among these different species. The aim of this study was to develop an analytical method to determine the presence of the neonicotinoid clothianidin in the Brazilian native stingless bee Tetragonisca angustula (local common name: Jataí). The method used for the chemical analysis involved a QuEChERS technique combined with UHPLC-MS/MS analysis. The developed method was subsequently used to analyze collected field samples. In addition, the acute toxicity of the pesticide to T. angustula was evaluated in a laboratory bioassay evaluating both lethal and sublethal endpoints. The analytical method was successfully developed with detection and quantification limits of 1.55 and 5 µg L-1, respectively, along with a linear range of 1-5 ng mL-1. Clothianidin was detected in environmental samples (9.2-32.9 ng g-1), and the exposure experiments demonstrated acute oral toxicity to adults of T. angustula, (24 h-LD50 of 0.16 ng a.i./bee), as well as no significative interference in acetylcholinesterase activity. Considering the obtained toxicity endpoints for T. angustula and those reported in the literature for other bee species, this study revealed that T. angustula is more (lethally) sensitive to clothianidin than other bee species, including those commonly used in environmental risk assessment studies. This thus also supports the call for using native test species in (regional) risk assessment evaluations.

Abamectin and difenoconazole monitoring in strawberry flowers and pollen sampled from Tetragonisca angustula (Latreille) (Hymenoptera: Apidae) hives located in crop vicinities.

The increase in agricultural productivity associated with the emergence and the extensive use of pesticides is undeniable. However, strong evidence indicates that this continuous demand is causing serious environmental impacts and bringing toxic effects to associated biota as pollinating insects. The present work aims the determination of the insecticide abamectin (ABA) and the fungicide difenoconazole (DIF) in strawberry flowers (Fragaria x ananassa DUCH.) and pollen sampled from beehives of the stingless bee Tetragonisca angustula Latreille (Hymenoptera: Apidae) located nearby strawberry fields. For analysis, QuEChERS method was optimized, and the analytical performance of those two pesticides was verified. Then, the method was applied to strawberry flowers and the pollen was sampled during three field campaigns. While abamectin was not detected, the systemic fungicide difenoconazole was determined in almost all flowers and pollen samples, demonstrating the major persistence of this pesticide in investigated matrices. The results were then discussed about the difenoconazole application rate and transport to colonies to estimate a preliminary environmental risk assessment for stingless native bees. All calculations were proceeded considering exposure rates and toxicity data from the literature, adapted from Apis mellifera studies. In this sense, the determination, application, and discussion about risk assessment figure out as an important tool to the knowledge about the preliminary risks of native bees exposed to pesticides.

Acute toxicity of the insecticide abamectin and the fungicide difenoconazole (individually and in mixture) to the tropical stingless bee Melipona scutellaris.

Stingless bees have been recognized as essential plant pollinators and producers of various natural products in neotropical areas. Research into the potential risks of pesticides they may be exposed to in agricultural fields, however, remains meagre. Especially the toxicity of pesticide mixtures likely to occur under real-world conditions and that are likely to exert synergetic effects has been poorly studied. The aim of the present study was therefore to evaluate the single and mixture acute contact and oral toxicity of commercial products containing the insecticide abamectin and the fungicide difenoconazole in laboratory bioassays with the Brazilian native stingless bee Melipona scutellaris. In addition, a comparison of the insecticide sensitivity of stingless bees relative to the honeybee Apis mellifera was made based on previously published toxicity data. Except for oral exposure to abamectin, M. scutellaris appeared to be more sensitive that A. mellifera in the single compound toxicity tests. A difenoconazole concentration at the NOEC (no observed effect concentration) level indicated a synergetic toxic interaction with abamectin. A sensitivity comparison based on published toxicity data for A. mellifera and stingless bees indicated several insecticidal modes of action having a high relative sensitivity to stingless bees that need especial consideration in future studies. The research findings highlight the need for testing native bee species and environmentally relevant pesticide mixtures in risk assessments to avoid underestimation of potential risks to bee populations and the subsequent loss of pollination ecosystem services.