Various studies have shown that bee-collected pollen sold as nutritional supplements may contain toxic pyrrolizidine alkaloids (PAs) and, thus, pose a potential health risk for consumers. The level of contamination may vary according to its geographical and botanical origin. Here, the PA content of pollen produced in Switzerland was studied and 32 commercially available bee-collected pollen supplements produced between 2010 and 2014 were analysed. In addition, at what time period bees collect PA-containing pollen was investigated. Hence, this study looked into the occurrence of PAs in pollen samples collected daily during two-to-three consecutive seasons. Furthermore, the PA spectrum in pollen was compared to the spectrum found in flower heads of PA-plants to unambiguously identify plants responsible for PA contamination of pollen. The PA concentration of commercial and daily collected pollen was determined by target analysis using an HPLC-MS/MS system, allowing the detection of 18 different PAs and PA N-oxides found in the genera Echium, Eupatorium and Senecio, while the comparison of the PA spectrum in pollen and flower heads was performed by LC-HR-MS, allowing the detection of all PA types in a sample, including saturated, non-carcinogenic PAs. Of the commercially available pollen, 31% contained PAs with a mean concentration of 319 ng/g, mainly Echium- and Eupatorium-type PAs, while the PA concentrations were below the limit of quantitation (LOQ) in 69% of the pollen samples. Bees collected pollen containing Echium-type PAs mainly in June and July, while they gathered pollen containing Eupatorium-type PAs from mid-July to August. Senecio-type PAs appeared from June to September. Comparison of the PA array in pollen and plants identified E. vulgare and E. cannabinum as the main plants responsible for PA contamination of Swiss bee-collected pollen, and to a lesser extent also identified plants belonging to the genus Senecio.

We studied the distribution of coumaphos in beeswax after a single application of CheckMite® to 15 bee colonies in the fall according to the manufacturer's instruction. Immediately after the treatment, residue levels were especially elevated in the wax that came into contact with the CheckMite® strips. During the following spring season, coumaphos levels ranged from 36 to 159 mg/kg in the wax of the brood frames next to the CheckMite® strips, and residue levels were about 10 times lower in the wax of frames that did not come into contact with the strips. Beeswax that was newly constructed 5 months after removing the treatment strips contained up to 7.3 mg/kg of coumaphos, and up to 0.5 and 2.3 mg/kg was detected in the honeycomb and capping wax, respectively. These results suggest that beeswax exposed to CheckMite® should not be recycled in order to prevent elevated concentrations of coumaphos in new foundations and hence to prevent honeybee larvae from being exposed to detrimental residue levels.beeswax / coumaphos / CheckMite® / honeybees / Apis mellifera / Varroa destructor

For the development of novel selection traits in honey bees, applicability under field conditions is crucial. We thus evaluated two novel traits intended to provide resistance against the ectoparasitic mite Varroa destructor and to allow for their straightforward implementation in honey bee selection. These traits are new field estimates of already-described colony traits: brood recapping rate (‘Recapping’) and solidness (‘Solidness’). ‘Recapping’ refers to a specific worker characteristic wherein they reseal a capped and partly opened cell containing a pupa, whilst ‘Solidness’ assesses the percentage of capped brood in a predefined area. According to the literature and beekeepers’ experiences, a higher recapping rate and higher solidness could be related to resistance to V. destructor. During a four-year field trial in Switzerland, the two resistance traits were assessed in a total of 121 colonies of Apis mellifera mellifera. We estimated the repeatability and the heritability of the two traits and determined their phenotypic correlations with commonly applied selection traits, including other putative resistance traits. Both traits showed low repeatability between different measurements within each year. ‘Recapping’ had a low heritability (h2 = 0.04 to 0.05, depending on the selected model) and a negative phenotypic correlation to non-removal of pin-killed brood (r = −0.23). The heritability of ‘Solidness’ was moderate (h2 = 0.24 to 0.25) and did not significantly correlate with resistance traits. The two traits did not show an association with V. destructor infestation levels. Further research is needed to confirm the results, as only a small number of colonies was evaluated.

The synthesis via in situ polymerization and characterization of nanocomposites (NCs) made from silica (SiO(2)) nanoparticles in a methyl methacrylate (MMA) monomer matrix is reported. We first investigate the transfer of well-defined spherical silica nanoparticles (NPs) (average radius R = 24.2 ± 3.2 nm) into the monomer solvent. We study the influence of two transfer methods and different surface chemistries on the resulting colloidal stability. The first transfer method consists of drying the silica particles into powder before dispersing them via ultrasonication in the MMA matrix. The second is based on repetitive centrifugations to exchange NPs from their synthetic milieu to pure MMA, avoiding the dried powder state. These transfer methods are compared for two kinds of NP surface chemistry, natural silanol (Si-OH) groups of the silica NPs and an additional silane coupling agent, namely 3-(trimethoxysilyl)propyl methacrylate (TPM), which mimics the monomeric group of the MMA dispersing milieu. We then characterize the morphology of the resulting nanocomposites prepared via in situ polymerization at a fixed amount of dispersed NPs (2.2 wt%; 1 vol%) using a combination of transmission electron microscopy (TEM) from ultramicrotomed nanocomposite films, thermogravimetry (TGA) and small angle X-ray scattering (SAXS).

Pesticide contamination of bee products is a widespread phenomenon. Due to its composition, bee bread is affected by both lipophilic and hydrophilic substances. As proof of concept of a monitoring campaign and to better understand the extent of contamination, we developed an analytical method based on a modified QuEChERS extraction, with subsequent separation by liquid chromatography and detection by mass spectrometry. This allowed for the quantitation of 51 agricultural- or beekeeping-associated pesticides in bee bread. The workflow was applied to 60 samples taken biweekly throughout spring to autumn 2022 from five colonies at a Swiss apiary in an agricultural area. In total, 30 pesticides were identified (> LOD), among which 26 pesticides were quantitated. The total number of pesticides detected per colony ranged from 11 to 19. The most prevalent substances (> LOQ) were two neonicotinoid insecticides, acetamiprid and thiacloprid (max. 16 μg/kg and 37 μg/kg, respectively); seven fungicides, azoxystrobin (max. 72 μg/kg), boscalid (max. 50 μg/kg), cyprodinil (max. 1965 μg/kg), difenoconazole (max. 73 μg/kg), mandipropamid (max. 33 μg/kg), pyraclostrobin (max. 8 μg/kg) and trifloxystrobin (max. 38 μg/kg); and two herbicides, prosulfocarb (max. 38 μg/kg) and terbuthylazine (max. 26 μg/kg). The study revealed strong variability in pesticide occurrence and concentrations among colonies sampled at the same site and date. The applied biweekly sampling of bee bread from March to August was shown to be reliable in capturing peak contaminations and revealing the onset of certain pesticides in bee bread. The study provides an adequate practical approach for pesticide monitoring campaigns.

Coumaphos is one of the most frequently detected pesticides in recycled beeswax. The objective was to assess the maximal level of coumaphos in foundation sheets that could exist without lethal effects on the honey bee larvae. Brood development was followed in cells drawn on foundation squares containing coumaphos ranging from 0 to 132 mg/kg. Furthermore, larval exposure was determined by measuring the coumaphos level in the drawn cells. Coumaphos levels in the initial foundation sheets up to 62 mg/kg did not increase brood mortality because the emergence rates of bees raised on these foundation squares were similar to controls (median of 51%). After a single brood cycle, coumaphos levels in the drawn cells were up to three times lower than the initial levels in foundation sheets. Hence, coumaphos levels of 62 mg/kg in the initial foundation sheets, almost the highest exposures, resulted in levels of 21 mg/kg in drawn cells. A significantly reduced emergence rate (median of 14%) was observed for bees raised on foundation sheets with initial coumaphos levels of 132 mg/kg, indicating increased brood mortality. Such levels resulted in coumaphos concentrations of 51 mg/kg in drawn cells, which is close to the median lethal concentration (LC50) as determined in previous in vitro experiments. In conclusion, brood mortality was increased on wax foundation sheets with initial coumaphos levels of 132 mg/kg, while no elevated mortality was observed for levels up to 62 mg/kg. Environ Toxicol Chem 2023;42:1816–1822. © 2023 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.

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