A new exposure protocol adapted for wild bees reveals species-specific impacts of the sulfoximine insecticide sulfoxaflor.

Wild bees are crucial pollinators of flowering plants and concerns are rising about their decline associated with pesticide use. Interspecific variation in wild bee response to pesticide exposure is expected to be related to variation in their morphology, physiology, and ecology, though there are still important knowledge gaps in its understanding. Pesticide risk assessments have largely focussed on the Western honey bee sensitivity considering it protective enough for wild bees. Recently, guidelines for Bombus terrestris and Osmia bicornis testing have been developed but are not yet implemented at a global scale in pesticide risk assessments. Here, we developed and tested a new simplified method of pesticide exposure on wild bee species collected from the field in Belgium. Enough specimens of nine species survived in a laboratory setting and were exposed to oral and topical acute doses of a sulfoximine insecticide. Our results confirm significant variability among wild bee species. We show that Osmia cornuta is more sensitive to sulfoxaflor than B. terrestris, whereas Bombus hypnorum is less sensitive. We propose hypotheses on the mechanisms explaining interspecific variations in sensitivity to pesticides. Future pesticide risk assessments of wild bees will require further refinement of protocols for their controlled housing and exposure.

Results of Ring-Testing of a Semifield Study Design to Investigate Potential Impacts of Crop Protection Products on Bumblebees (Hymenoptera, Apidae) and a Proposal of a Potential Test Design.

In Europe, the risk assessment for bees at the European Union or national level has always focussed on potential impacts on honeybees. During the revision of the European Food Safety Authority bee guidance it was explicitly stated that bumblebees and solitary bees should be considered as well and consequently concerns were raised regarding the representativeness of honeybees for these other bee species. These concerns originate from differences in size as well as differences in behavioral and life history traits of other bee species. In response to this concern, the non-Apis working group of the International Commission for Plant-Pollinator Relationships initiated a ring-test of a semifield tunnel study design using the bumblebee Bombus terrestris. Nine laboratories participated, validating and improving the proposed design over a 2-year period. The intention of the ring-test experiments was to develop and if possible, establish a test protocol to conduct more standardized semifield tests with bumblebees. In the present study, the results of the ring-tests are summarized and discussed to give recommendations for a promising experimental design. Environ Toxicol Chem 2022;41:2548-2564. © 2022 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.

Acute oral toxicity and risks of four classes of systemic insecticide to the Common Eastern Bumblebee (Bombus impatiens).

The Common Eastern Bumblebee (Bombus impatiens) is native to North America with an expanding range across Eastern Canada and the USA. This species is commercially produced primarily for greenhouse crop pollination and is a common and abundant component of the wild bumblebee fauna in agricultural, suburban and urban landscapes. However, there is a dearth of pesticide toxicity information about North American bumblebees. The present study determined the acute oral lethal toxicity (48-h LD50) of: the butenolide, flupyradifurone (>1.7 µg/bee); the diamide, cyantraniliprole (>0.54 µg/bee); the neonicotinoid, thiamethoxam (0.0012 µg/bee); and the sulfoximine, sulfoxaflor (0.0177 µg/bee). Compared with published honey bee (Apis mellifera) LD50 values, the present study shows that sulfoxaflor and thiamethoxam are 8.3× and 3.3× more acutely toxic to B. impatiens, whereas flupyradifurone is more acutely toxic to A. mellifera. The current rule of thumb for toxicity extrapolation beyond the honey bee as a model species, termed 10× safety factor, may be sufficient for bumblebee acute oral toxicity. A comparison of five risk assessment equations suggested that the Standard Risk Approach (SRA) and Fixed Dose Risk Approach (FDRA) provide more nuanced levels of risk evaluation compared to the Exposure Toxicity Ratio (ETR), Hazard Quotient (HQ), and Risk Quotient (RQ), primarily because the SRA and FDRA take into account real world variability in pollen and nectar pesticide residues and the chances that bees may be exposed to them.

Results of 2-Year Ring Testing of a Semifield Study Design to Investigate Potential Impacts of Plant Protection Products on the Solitary Bees Osmia Bicornis and Osmia Cornuta and a Proposal for a Suitable Test Design.

There are various differences in size, behavior, and life history traits of non-Apis bee species compared with honey bees (Apis mellifera; Linnaeus, 1758). Currently, the risk assessment for bees in the international and national process of authorizing plant protection products has been based on honey bee data as a surrogate organism for non-Apis bees. To evaluate the feasibility of a semifield tunnel test for Osmia bicornis (Linnaeus, 1758) and Osmia cornuta (Latreille, 1805), a protocol was developed by the non-Apis working group of the International Commission for Plant-Pollinator Relationships, consisting of experts from authorities, academia, and industry. A total of 25 studies were performed over a 2-yr period testing a replicated control against a replicated positive control using either a dimethoate or diflubenzuron treatment. Studies were regarded to be valid, if ≥30% of released females were found to occupy the nesting units in the night/morning before the application (establishment). Thirteen studies were regarded to be valid and were analyzed further. Parameters analyzed were nest occupation, flight activity, cell production (total and per female), cocoon production (total and per female), emergence success, sex ratio, and mean weight of females and males. Dimethoate was a reliable positive control at the tested rate of 75 g a.i./ha, once >30% females had established, displaying acute effects such as reduction in flight activity, increase in adult mortality (shown by nest occupation), and reproduction ability of the females (total cell and cocoon production). On the other hand, no effects on larval and pupal development were observed. The growth regulator diflubenzuron had statistically significant effects on brood development, causing mortality of eggs and larvae at a rate of approximately 200 g a.i./ha, whereas fenoxycarb did not cause any significant effects at the tested rates of 300 and 600 g a.i./ha. In conclusion, the ring-test protocol proved to be adequate once the study comprised a well-established population of female Osmia bees, and the results improved in the second year as the laboratories increased their experience with the test organism. It is noted that the success of a study strongly depends on the experience of the experimenter, the crop quality, the quality of the cocoons, and the weather conditions. Based on these finding, recommendations for a semifield study design with Osmia spp. are proposed. Environ Toxicol Chem 2021;40:236-250. © 2020 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.

Possible Spillover of Pathogens between Bee Communities Foraging on the Same Floral Resource.

Viruses are known to contribute to bee population decline. Possible spillover is suspected from the co-occurrence of viruses in wild bees and honey bees. In order to study the risk of virus transmission between wild and managed bee species sharing the same floral resource, we tried to maximize the possible cross-infections using Phacelia tanacetifolia, which is highly attractive to honey bees and a broad range of wild bee species. Virus prevalence was compared over two years in Southern France. A total of 1137 wild bees from 29 wild bee species (based on COI barcoding) and 920 honey bees (Apis mellifera) were checked for the seven most common honey bee RNA viruses. Halictid bees were the most abundant. Co-infections were frequent, and Sacbrood virus (SBV), Black queen cell virus (BQCV), Acute bee paralysis virus (ABPV) and Israeli acute paralysis virus (IAPV) were widespread in the hymenopteran pollinator community. Conversely, Deformed wing virus (DWV) was detected at low levels in wild bees, whereas it was highly prevalent in honey bees (78.3% of the samples). Both wild bee and honey bee virus isolates were sequenced to look for possible host-specificity or geographical structuring. ABPV phylogeny suggested a specific cluster for Eucera bees, while isolates of DWV from bumble bees (Bombus spp.) clustered together with honey bee isolates, suggesting a possible spillover.

Whole-Body Acute Contact Toxicity of Formulated Insecticide Mixtures to Blue Orchard Bees (Osmia lignaria).

Blue orchard bees, [Osmia lignaria (Say) (Hymenoptera: Megachilidae)], have been developed as an important pollinator for orchard crops in North America over the last 40 years. The toxicity of several pesticides to O. lignaria and other Osmia species has been previously reported. However, the field-realistic toxicity of formulated premix insecticides comprised of multiple active ingredients (each with a different mode of action) to O. lignaria has not been assessed. Here, we use a customized spray tower in a laboratory setting to assess adult male and female whole-body direct contact exposure to four formulated pesticide mixtures: thiamethoxam + lambda-cyhalothrin (TLC), imidacloprid + beta-cyfluthrin (IBC), chlorantraniliprole + lambda-cyhalothrin (CLC) and methoxyfenozide + spinetoram (MS) by directly spraying anesthetized bees in Petri dishes. Separately, adult male and female whole-body direct contact exposure to formulated imidacloprid (I), beta-cyfluthrin (BC) and their 1:1 binary combination (IBC) was assessed using the same experimental method. Resulting mortality in each study was screened up to 96 h post-treatment to determine acute whole-body contact toxicity. In the first study, TLC and IBC resulted in statistically higher mortality at 24 and 48 h than the two other insecticide combinations tested. The CLC and MS combinations were slower acting and the highest mortality for O. lignaria exposed to these mixtures was recorded at 96 h. We did observe significant differences in toxicity between CLC and MS. In the second study, exposure to the 1:1 binary combination of IBC caused overall significantly higher mortality than exposure to I or BC alone. Both active ingredients alone, however, demonstrated equivalent levels of mortality to the 1:1 binary combination treatment at the 96 h observation reading, indicating increased speed of kill, but not necessarily increased toxicity. Significant differences in the onset of mortality following acute contact whole-body exposure to the formulated insecticide mixtures and individual active ingredients tested were consistently observed across all experiments in both studies.

Bee Viruses: Routes of Infection in Hymenoptera.

Numerous studies have recently reported on the discovery of bee viruses in different arthropod species and their possible transmission routes, vastly increasing our understanding of these viruses and their distribution. Here, we review the current literature on the recent advances in understanding the transmission of viruses, both on the presence of bee viruses in Apis and non-Apis bee species and on the discovery of previously unknown bee viruses. The natural transmission of bee viruses will be discussed among different bee species and other insects. Finally, the research potential of in vivo (host organisms) and in vitro (cell lines) serial passages of bee viruses is discussed, from the perspective of the host-virus landscape changes and potential transmission routes for emerging bee virus infections.

Are honeybees suitable surrogates for use in pesticide risk assessment for non-Apis bees?

Historically, bee regulatory risk assessment for pesticides has centred on the European honeybee (Apis mellifera), primarily due to its availability and adaptability to laboratory conditions. Recently, there have been efforts to develop a battery of laboratory toxicity tests for a range of non-Apis bee species to directly assess the risk to them. However, it is not clear whether the substantial investment associated with the development and implementation of such routine screening will actually improve the level of protection of non-Apis bees. We argue, using published acute toxicity data from a range of bee species and standard regulatory exposure scenarios, that current first-tier honeybee acute risk assessment schemes utilised by regulatory authorities are protective of other bee species and further tests should be conducted only in cases of concern. We propose similar analysis of alternative exposure scenarios (chronic and developmental) once reliable data for non-Apis bees are available to expand our approach to these scenarios. In addition, we propose that in silico (simulation) approaches can then be used to address population-level effects in more field-realistic scenarios. Such an approach could lead to a protective, but also workable, risk assessment for non-Apis species while contributing to pollination security in agricultural landscapes around the globe. © 2019 Society of Chemical Industry.

Potential surrogate plants for use in semi-field pesticide risk assessment with Megachile rotundata.

BACKGROUND: Current regulatory pesticide risk assessments for bees are based primarily on the honey bee (Apis mellifera) and may not always be protective of solitary bees. To incorporate solitary bees into the risk assessment process, standardized methods to assess the hazard of pesticides under semi-field (Tier II) conditions will be needed. We conducted a series of experiments over 2 years to assess potential surrogate plants and adult release rates for use in semi-field experiments with the alfalfa leafcutting bee (ALB, Megachile rotundata). METHODS: We compared ALB foraging activity and reproduction on 12 m2 plots of flowering alfalfa (Medicago sativa) and buckwheat (Fagopyrum esculentum) at low (10♀/20♂) and high (20♀/40♂) adult release rates. The following year, we assessed the same endpoints on plots of purple tansy (Phacelia tanacetifolia) at a release rate of 10♀/15♂. RESULTS: Although ALB foraging activity was high on buckwheat plots, fewer adults were produced compared to alfalfa plots. On alfalfa, there were no differences in foraging activity, nesting, or reproduction between the low and high release rates. ALB readily foraged from purple tansy flowers, but females avoided purple tansy leaves for leaf cell construction. DISCUSSION: Our study suggests that buckwheat alone cannot support ALB during semi-field studies on small plots. For alfalfa, we recommend a maximum release rate of 10♀/20♂ in 12 m2 plots. Further study of higher ALB release rates on purple tansy is warranted. A mixed planting of purple tansy and a plant suitable for leaf piece collection (e.g., buckwheat) may provide favorable conditions for ALB activity and reproduction during semi-field testing.