Benaki Phytopathological Institute, Department of Pesticides Control and Phytopharmacy, Laboratory of Pesticides' Toxicology, 8 St. Delta Street, Kifissia, 14561 Athens, Greece.
Benaki Phytopathological Institute, Department of Pesticides Control and Phytopharmacy, Laboratory of Pesticides' Toxicology, 8 St. Delta Street, Kifissia, 14561 Athens, Greece.
Sci Total Environ. 2021 May 15;769:145213. doi: 10.1016/j.scitotenv.2021.145213. Epub 2021 Jan 17.
The aim of this study was to investigate reported cases of honeybee mortality incidents and the potential association to pesticide exposure and to their metabolites. The same honeybee samples were also assessed for Varroa mites, and Nosema microsporidia provoked infections to provide an integrated picture of all observable stressors that may impact bees' survival. Thus, honeybee samples from different areas of Greece (2014-2018) were analyzed for the presence of pesticide residues and metabolites. In this context, an existing LC-ESI-QqQ-MS multiresidue method of analytes of different chemical classes such as neonicotinoids, organophosphates, triazoles, carbamates, was enriched with additional active substances, developed and validated. A complementary GC-EI-QqQ-MS method was also exploited for the same scope covering pyrethroid compounds. Both methods monitored more than 150 active substances and metabolites and presented acceptable linearity over the ranges assayed. The calculated recoveries ranged from 65 to 120% for the three concentration levels, while the precision (RSD%) values ranged between 4 and 15%. Therefore, this approach proved sufficient to act as a monitoring tool for the determination of pesticide residues in cases of suspected honeybee poisoning incidents. From the analysis of 320 samples, the presence of 70 active substances and metabolites was confirmed with concentrations varying from 1.4 ng/g to 166 μg/g. Predominant detections were the acaricide coumaphos, several neonicotinoids exemplified by clothianidin, organophosporous compounds dimethoate and chlorpyrifos, and some pyrethroids. Metabolites of imidacloprid, chlorpyrifos, coumaphos, acetamiprid, fenthion and amitraz were also identified. Concerning Nosema and Varroa they were identified in 27 and 22% of samples examined, respectively, verifying their prevalence and coexistence with pesticides and their metabolites in honeybees.
本研究旨在调查蜜蜂死亡事件的报告案例，并探讨其与农药暴露及其代谢物的潜在关联。还评估了相同的蜜蜂样本中的瓦螨和微孢子虫感染，以提供所有可能影响蜜蜂生存的可观察应激因素的综合情况。因此，分析了来自希腊不同地区（2014-2018 年）的蜜蜂样本，以检测农药残留和代谢物的存在。在这种情况下，用额外的活性物质丰富、开发和验证了一种现有的 LC-ESI-QqQ-MS 多残留分析方法，用于分析不同化学类别的分析物，如新烟碱类、有机磷类、三唑类、氨基甲酸酯类。还利用互补的 GC-EI-QqQ-MS 方法对同样的范围进行了研究，涵盖了拟除虫菊酯化合物。这两种方法都监测了 150 多种活性物质和代谢物，在测定的范围内具有可接受的线性。计算的回收率在三个浓度水平下为 65%至 120%，而精密度（RSD%）值在 4%至 15%之间。因此，这种方法足以作为怀疑蜜蜂中毒事件中农药残留测定的监测工具。对 320 个样本的分析证实了 70 种活性物质和代谢物的存在，浓度从 1.4ng/g 到 166μg/g 不等。主要检测到杀螨剂残杀威、几种新烟碱类化合物，如噻虫嗪、有机磷化合物甲拌磷和毒死蜱，以及一些拟除虫菊酯类化合物。还鉴定了吡虫啉、毒死蜱、残杀威、噻虫嗪、三唑磷、丙硫磷和咪鲜胺的代谢物。关于微孢子虫和瓦螨，在所检查的样本中分别有 27%和 22%被识别，这验证了它们在蜜蜂中的流行率以及与农药及其代谢物的共存。
