Design and planning of a transdisciplinary investigation into farmland pollinators: rationale, co-design, and lessons learned

To provide a complete portrayal of the multiple factors negatively impacting insects in agricultural landscapes it is necessary to assess the concurrent incidence, magnitude, and interactions among multiple stressors over substantial biogeographical scales. Trans-national ecological field investigations with wide-ranging stakeholders typically encounter numerous challenges during the design planning stages, not least that the scientific soundness of a spatially replicated study design must account for the substantial geographic and climatic variation among distant sites. ‘PoshBee’ (Pan-European assessment, monitoring, and mitigation of Stressors on the Health of Bees) is a multi-partner transdisciplinary agroecological project established to investigate the suite of stressors typically encountered by pollinating insects in European agricultural landscapes. To do this, PoshBee established a network of 128 study sites across eight European countries and collected over 50 measurements and samples relating to the nutritional, toxicological, pathogenic, and landscape components of the bees’ environment. This paper describes the development process, rationale, and end-result of each aspect of the of the PoshBee field investigation. We describe the main issues and challenges encountered during the design stages and highlight a number of actions or processes that may benefit other multi-partner research consortia planning similar large-scale studies. It was soon identified that in a multi-component study design process, the development of interaction and communication networks involving all collaborators and stakeholders requires considerable time and resources. It was also necessary at each planning stage to be mindful of the needs and objectives of all stakeholders and partners, and further challenges inevitably arose when practical limitations, such as time restrictions and labour constraints, were superimposed upon prototype study designs. To promote clarity for all stakeholders, for each sub-component of the study, there should be a clear record of the rationale and reasoning that outlines how the final design transpired, what compromises were made, and how the requirements of different stakeholders were accomplished. Ultimately, multi-national agroecological field studies such as PoshBee benefit greatly from the involvement of diverse stakeholders and partners, ranging from field ecologists, project managers, policy legislators, mathematical modelers, and farmer organisations. While the execution of the study highlighted the advantages and benefits of large-scale transdisciplinary projects, the long planning period emphasized the need to formally describe a design framework that could facilitate the design process of future multi-partner collaborations

Method of Glyphosate, AMPA, and Glufosinate Ammonium Determination in Beebread by Liquid Chromatography—Tandem Mass Spectrometry after Molecularly Imprinted Solid-Phase Extraction

The aim of this study was to develop a method for the determination of glyphosate, its metabolite aminomethylphosphonic acid (AMPA), and glufosinate ammonium residues in beebread samples, which could then be used to assess bees’ exposure to their residues. The complexity of beebread’s matrix, combined with the specific properties of glyphosate itself, required careful selection and optimization of each analysis step. The use of molecularly imprinted solid-phase extraction (MIP-SPE) by AFFINIMIP glyphosate as an initial clean-up step significantly eliminated matrix components and ensured an efficient derivatization step. Colorless beebread extracts were derivatized by the addition of 9-fluorenylmethyl chloroformate (FMOC-Cl). After derivatization, in order to remove FMOC-OH and residual borate buffer, a solid-phase extraction (SPE) clean-up step using Oasis HLB was carried out. Instrumental analysis was performed by liquid chromatography coupled with tandem mass spectrometry (LC–MS/MS). The method was validated according to the SANTE/11312/2021 guideline at concentrations of 5, 10, and 100 µg/kg, and satisfactory recovery (trueness) values (76–111%) and precision (RSDr) ≤ 18% were obtained. The limit of quantification (LOQ) was 5 µg/kg for AMPA and glufosinate ammonium and 10 µg/kg for glyphosate. The method was positively verified by the international proficiency test. Analysis of beebread samples showed the method’s usefulness in practice. The developed method could be a reliable tool for the assessment of beebread’s contamination with residues of glyphosate, its metabolite AMPA, and glufosinate ammonium

Pesticide Poisoning of Honeybees: A Review of Symptoms, Incident Classification, and Causes of Poisoning

During the 2000s, the problem of pesticide poisoning of honeybees seemed to be almost solved. The number of cases has decreased in comparison to the 1970s. The problem of acute honeybee poisoning, however, has not disappeared, but instead has transformed into a problem of poisoning from ‘traditional’ pesticides like organophosphorus pesticides or pyrethroids, to poisoning from additional sources of ‘modern’ systemic neonicotinoids and fipronil. In this article, the biological activity of pesticides was reviewed. The poisoning symptoms, incident definitions, and monitoring systems, as well as the interpretation of the analytical results, were also reviewed. The range of pesticides, and the detected concentrations of pesticides in poisoned honeybee samples, were reviewed. And, for the first time, cases of poisoning related to neonicotinoids were reviewed. The latter especially is of practical importance and could be helpful to analysts and investigators of honeybee poisoning incidents. It is assumed that secondary poisoning induced by plant collected materials contaminated with systemic pesticides occurs. Food stored in a hive and contaminated with systemic pesticides consumed continuously by the same generation of winter bees, may result in sub-lethal intoxication. This leads to abnormal behaviour identified during acute intoxication. The final result is that the bees discontinue their social role in the honeybee colony super organism, and colony collapse disorder (CCD) takes place. The process described above refers primarily to robust and strong colonies that were able to collect plenty of food due to effective plant protection

Development of a High Performance Liquid Chromatography with Diode Array Detector (HPLC-DAD) Method for Determination of Biogenic Amines in Ripened Cheeses

Biogenic amines (BAs) are organic, basic nitrogenous compounds formed during the decarboxylation of amino acids. A method for the determination of eight biogenic amines (tryptamine, 2-phenyletylamine, putrescine, cadaverine, histamine, tyramine, spermidine, spermine) in ripened cheeses was developed and validated. Cheese samples with the addition of internal standards were extracted with 0.2 M perchloric acid and pre-column derivatized with dansyl chloride at 60 °C for 15 min, purified with toluene and dried under a stream of nitrogen. The samples were analyzed using high performance liquid chromatography with diode array detector (HPLC-DAD). The method was validated with the BAs at three concentration levels: 50, 100, and 200 mg/kg, respectively. The obtained values of correlation coefficient (R2) ranged at 0.9997–0.9998 for all of compounds. The limits of detection (LOD) and quantification (LOQ) were in ranges 1.53–1.88 and 5.13–6.28 mg/kg, respectively. The recovery for all of biogenic amines ranged from 70 to 120% and the precision (RSDr) value were <20%. The validated method was applied to analysis of 35 real ripened cheese samples purchased in Poland

Amitraz marker residues in honey from honeybee colonies treated with Apiwarol

Amitraz is a formamide exhibiting both acaricidal and insecticidal activity and is frequently used by beekeepers to protect honeybee colonies against Varroa destructor mites. The aim of this apiary trial was to evaluate the impact of honeybee colony fumigation with amitraz on the level of contamination of honey stored in combs

L'étude de néonicotinoides en culture betteravière

Des chercheurs de l'UCLouvain identifie une menace pour les pollinisateur

Are intercropping cover crops a potential threat for pollinators due to neonicotinoid residues in floral resources?

Intercropping cover crops have become mandatory in areas at risk of nitrogen leaching to groundwater. These covers include several attractive late-flowering entomophilous species. They can therefore represent crucial floral resources (pollen and nectar) for pollinating insects in early autumn. Pesticides used in previous crops, however, represent a potential risk for pollinators when they are transferred to the intercropping cover plants and their floral resources. We studied the potential transfer of clothianidin (a neonicotinoid insecticide), applied two years earlier in a beet cultivation, from soil to plants and to the floral resources of three common cover species: Phacelia tanacetifolia, Sinapis alba, and Vicia faba. Soils, entire plants, flowers, and nectar were collected from plants grew in greenhouses, and soils and pollen were collected on a treated field. Our results showed that clothianidin was still present in soils (4.5 ng g−1). The residues accumulated in plants (5-15 times higher concentrations than in soils) and were present in pollen of both Vicia faba (0.07 ng g−1) and Sinapis alba (1.7 ng g−1) and in nectar of both Sinapis alba and Phacelia tanacetifolia

Visitation of <i>Apis mellifera</i> L. in Runner Bean (<i>Phaseolus coccineus</i> L.) and Its Exposure to Seasonal Agrochemicals in Agroecosystems

Plant species and abiotic factors including season appear to be the most important variables influencing the frequency of visits by honeybees (Apis mellifera L.). In the present study, we evaluated the activity of honeybee workers visiting runner bean (Phaseolus coccineus L.) local cultivar ‘Piękny Jaś’. The runner beans are widely cultivated in south-eastern Poland, and are an important forage plant for honeybees in agroecosystems. We aimed at a comprehensive monitoring of the health of colonies and symptoms in A. mellifera in response to acute exposure to pesticides. The most numerous visits of A. mellifera were observed at the highest flower opening of the runner bean. A very weak positive correlation was observed between the number of honeybees on P. coccineus, the number of visited flowers, the time spent per flower and air temperature. The visitation rates of honeybees were more frequent at mid-day and decreased after 15:00. Signs of poisoning were detected in two out of seven apiaries monitored for acute pesticide exposure symptoms on runner bean plantations. The analysis of dead honeybee samples revealed the acute exposure of honeybees to the imidacloprid (neonicotinoid) and chlorpyrifos (organophosphorus) insecticides, which are highly toxic and banned in the European Union. Hazard quotient (HQ) screening showed an elevated burden of imidacloprid and chlorpyrifos corresponding to 7.1% and 10% of the LDD50, respectively, most likely indicating bee poisoning due to chronic exposure to these substances with contaminated food. Noteworthy was the presence of three fungicides that could pose a risk of poisoning in honeybees

Nutritional stress exacerbates impact of a novel insecticide on solitary bees' behaviour, reproduction and survival.

peer reviewedPesticide exposure and food stress are major threats to bees, but their potential synergistic impacts under field-realistic conditions remain poorly understood and are not considered in current pesticide risk assessments. We conducted a semi-field experiment to examine the single and interactive effects of the novel insecticide flupyradifurone (FPF) and nutritional stress on fitness proxies in the solitary bee Osmia bicornis. Individually marked bees were released into flight cages with monocultures of buckwheat, wild mustard or purple tansy, which were assigned to an insecticide treatment (FPF or control) in a crossed design. Nutritional stress, which was high in bees foraging on buckwheat, intermediate on wild mustard and low on purple tansy, modulated the impact of insecticide exposure. Within the first day after application of FPF, mortality of bees feeding on buckwheat was 29 times higher compared with control treatments, while mortality of FPF exposed and control bees was similar in the other two plant species. Moreover, we found negative synergistic impacts of FPF and nutritional stress on offspring production, flight activity, flight duration and flower visitation frequency. These results reveal that environmental policies and risk assessment schemes that ignore interactions among anthropogenic stressors will fail to adequately protect bees and the pollination services they provide.3729 - POSHBEE - Pan-European Assessment, Monitoring, and Mitigation of Of Stressors on the Health of BEEs - Sources publiques européenne