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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. [ABSTRACT FROM AUTHOR]

Insect pollination is an important process in the natural ecosystem and plays a vital role in crop reproduction. The bumblebee, Bombus terrestris (L.), is a widely used pollinator for crops, especially tomato. The efficiency of bee pollination is related to many factors, including biotic factors such as bee species, nutrition plants, and herbivores. Bemisia tabaci (Gennadius) (Homoptera: Aleyrodidae) is one of the most common and serious pests of tomato plants. However, whether B. tabaci affects the pollination behaviour of B. terrestris on tomato, and how plant–herbivore–pollinator interactions are unclear. In this study, we found that B. terrestris preferred B. tabaci-damaged plants. Furthermore, plant volatiles of tomato plants damaged by B. tabaci were detected and analysed. Electroantennogram (EAG) and the Y-tube bioassay were used for olfactory responses of B. terrestris to plant volatiles. Interestingly, methyl hexanoate and eugenol induced by B. tabaci in tomato plants were found to significantly attract B. terrestris. Therefore, Bemisia tabaci influenced B. terrestris preference through volatiles. The plant volatile-mediated interaction between B. tabaci and B. terrestris could enrich our understanding of the interaction between herbivores and pollinators. In addition, methyl hexanoate and eugenol could be applied for synergist of pollination. [ABSTRACT FROM AUTHOR]

Many pollinators, such as bees, hummingbirds and bats, use multi-destination routes (traplines) to exploit familiar plant resources. However, it is not clear to what extent the mechanisms underpinning trapline development and optimisation are comparable across species. Here we compared route formation, repeatability and efficiency by foragers of two social bee species, the solo foraging bumblebee Bombus terrestris and the mass foraging honey bee Apis mellifera, in the same laboratory conditions. In a simple routing task (with four artificial flowers), all bumblebees and honey bees developed a route, although honey bees were slower to do so. In a more complex routing task (with six flowers), however, only bumblebees developed a route between the 6 flowers. Honey bees took a longer time to discover all flowers and developed routes between fewer flowers. Comparing bumblebees and honey bees using the same experimental paradigm thus revealed key behavioural differences likely resulting from their contrasting collective foraging strategies. Significance statement: Pollinators face some of the most complex foraging challenge among animals: collecting small amounts of nectar from large numbers of flowers in a minimum of time. Previous studies report bees, birds and bats rely on spatial memory to develop routes optimising foraging efficiency. Here we compared this routing behaviour in two major pollinators, the buff-tailed bumblebee and the Western honey bee, in the same experimental conditions. We found that bumblebees were much faster and more efficient at visiting flowers following repeatable sequences than honey bees. This behavioural difference suggests bee cognition is shaped by their social ecology and illuminates about the known complementarity of different bee species for pollination. [ABSTRACT FROM AUTHOR]

In uncertain environments, animals often face the challenge of deciding whether to stay with their current foraging option or leave to pursue the next opportunity. The voluntary decision to persist at a location or with one option is a critical cognitive ability in animal temporal decision-making. Little is known about whether foraging insects form temporal expectations of reward and how these expectations affect their learning and rapid, short-term foraging decisions. Here, we trained bumblebees on a simple colour discrimination task whereby they entered different opaque tunnels surrounded by coloured discs (artificial flowers) and received reinforcement (appetitive sugar water or aversive quinine solution depending on flower colour). One group received reinforcement immediately and the other after a variable delay (0–3 s). We then recorded how long bees were willing to wait/persist when reinforcement was delayed indefinitely. Bumblebees trained with delays voluntarily stayed in tunnels longer than bees trained without delays. Delay-trained bees also waited/persisted longer after choosing the reward-associated flower compared to the punishment-associated flower, suggesting stimulus-specific temporal associations. Strikingly, while training with delayed reinforcement did not affect colour discrimination, it appeared to facilitate the generalisation of temporal associations to ambiguous stimuli in bumblebees. Our findings suggest that bumblebees can be trained to form temporal expectations, and that these expectations can be incorporated into their decision-making processes, highlighting bumblebees' cognitive flexibility in temporal information usage. Significance statement: The willingness to voluntarily wait or persist for potential reward is a critical aspect of decision-making during foraging. Investigating the willingness to persist across various species can shed light on the evolutionary development of temporal decision-making and related processes. This study revealed that bumblebees trained with delays to reinforcement from individual flowers were able to form temporal expectations, which, in turn, generalised to ambiguous stimuli. These findings contribute to our understanding of temporal cognition in an insect and the potential effects of delayed rewards on foraging behaviour. [ABSTRACT FROM AUTHOR]

Honeybees are well-established bioindicators for different types of pollutants. This study aims to establish another species of the Apidae family as a bioindicator, with a distinct behaviour and life cycle. The bumblebee Bombus terrestris was used as a bioindicator for 12 metals. Bumblebee hives were placed at sampling sites in and around the city of Aachen, Germany, and metal concentrations were assessed using ICP-MS. Metal concentrations were compared to those found in honeybees described in the literature. Spatial differences in metal patterns were investigated by comparing two land-use types: urban and agrarian. Seasonal differences were compared by taking samples in spring and summer. All analysed metals were detected above the detection limit and within or even above the concentration range found in honeybees. Significant spatial differences were found for the metalloid B and the metal Cd with higher concentrations at the agrarian sites than the urban sites. Significant seasonal differences were found for 8 metals: Fe, Zn, Cu, Ni, Cd, and As concentrations were higher in summer than in spring, while B and V concentrations were higher in spring. To categorise the results, we applied the honeybee contamination index (HCI) and adapted it to bumblebee purposes. According to the HCI, only one agrarian site showed a high contamination level. This study shows that bumblebees are suitable bioindicators for metals. The obtained data can serve as a first baseline in the establishment of additional monitoring studies or risk assessments. [ABSTRACT FROM AUTHOR]

The invasive hornet Vespa velutina nigrithorax is considered a proliferating threat to pollinators in Europe and Asia. While the impact of this species on managed honey bees is well-documented, effects upon other pollinator populations remain poorly understood. Nonetheless, dietary analyses indicate that the hornets consume a diversity of prey, fuelling concerns for at-risk taxa. Here, we quantify the impact of V. velutina upon standardised commercially-reared colonies of the European bumblebee, Bombus terrestris terrestris. Using a landscape-scale experimental design, we deploy colonies across a gradient of local V. velutina densities, utilising automated tracking to non-invasively observe bee and hornet behaviour, and quantify subsequent effects upon colony outcomes. Our results demonstrate that hornets frequently hunt at B. terrestris colonies, being preferentially attracted to those with high foraging traffic, and engaging in repeated—yet entirely unsuccessful—predation attempts at nest entrances. Notably however, we show that B. terrestris colony weights are negatively associated with local V. velutina densities, indicating potential indirect effects upon colony growth. Taken together, these findings provide the first empirical insight into impacts on bumblebees at the colony level, and inform future mitigation efforts for wild and managed pollinators. The influence of the invasive hornet Vespa velutina on Bombus terrestris colonies is quantified along a gradient of hornet densities, revealing negative impacts upon B. terrestris colony growth. [ABSTRACT FROM AUTHOR]

Many pollinators, including bumble bees, are in decline. Such declines are known to be driven by a number of interacting factors. Decreases in bee populations may also negatively impact the key ecosystem service, pollination, that they provide. Pesticides and parasites are often cited as two of the drivers of bee declines, particularly as they have previously been found to interact with one another to the detriment of bee health. Here we test the effects of an insecticide, sulfoxaflor, and a highly prevalent bumble bee parasite, Crithidia bombi, on the bumble bee Bombus terrestris. After exposing colonies to realistic doses of either sulfoxaflor and/or Crithidia bombi in a fully crossed experiment, colonies were allowed to forage on field beans in outdoor exclusion cages. Foraging performance was monitored, and the impacts on fruit set were recorded. We found no effect of either stressor, or their interaction, on the pollination services they provide to field beans, either at an individual level or a whole colony level. Further, there was no impact of any treatment, in any metric, on colony development. Our results contrast with prior findings that similar insecticides (neonicotinoids) impact pollination services, and that sulfoxaflor impacts colony development, potentially suggesting that sulfoxaflor is a less harmful compound to bee health than neonicotinoids insecticides. [ABSTRACT FROM AUTHOR]

Neonicotinoids harm especially bumblebees and other species in agricultural areas all over the world. The toxic effect of thiamethoxam from the neonicotinoid group has been little studied especially on bees. This research aimed to evaluate the effects of thiamethoxam on the immune system cells of Bombus terrestris workers. Different ratios of 1/1000, 1/100 and 1/10 of the recommended maximum application dose of thiamethoxam formed the experimental groups. Ten foraging workers were used for each dose and control groups. Contamination was ensured by spraying the prepared suspensions at different ratios to the bees for 20 s at a pressure of 1 atm. The effects of thiamethoxam on the structures of immune system cells of bumblebees and the amount of these cells were investigated after 48 h of exposure. In general, anomalies such as vacuolization, cell membrane irregularities and cell shape changes were detected in prohemocyte, plasmatocyte, granulocyte, spherulocyte and oenocytoid in each dose group. Hemocyte area measurements in all groups were examined comparatively between groups. In general, granulocyte and plasmatocyte sizes were decreased, while spherulocyte and oenocytoid were increased. It was also determined that there was a significant decrease in the amount of hemocytes in the 1 mm3 hemolymph as dose increased. The results of the study revealed that sublethal doses of thiamethoxam negatively affected hemocytes and their amounts of B. terrestris workers. [ABSTRACT FROM AUTHOR]

The rapid naturalization of Bombus terrestris across the Nemuro Peninsula has led to a decline in two closely related native Japanese species, namely Bombus hypocrita sapporensis and Bombus cryptarum florilegus, both belonging to the common subgenus Bombus. Although it is widely believed that cross-mating of native and non-native species is influenced by the common male sex pheromone in this region, no study has been conducted to substantiate this claim. Thus, we investigated the cross-activities of male sex pheromones between native and non-native bumblebees, as well as the frequencies of cross-mating, using chemical and DNA assays. Our gas chromatography–electroantennographic detector analyses and behavioral tests revealed the presence of sex pheromonal cross-activities between B. terrestris and the two Japanese bumblebees species. Furthermore, DNA analyses revealed the occurrence of cross-mating between native and non-native species in the Nemuro Peninsula. Overall, these results indicate the immediate need for conservation measures to safeguard Japanese bumblebee populations in the Nemuro Peninsula. [ABSTRACT FROM AUTHOR]

Pesticides are recognised as a key threat to pollinators, impacting their health in many ways. One route through which pesticides can affect pollinators like bumblebees is through the gut microbiome, with knock-on effects on their immune system and parasite resistance. We tested the impacts of a high acute oral dose of glyphosate on the gut microbiome of the buff tailed bumblebee (Bombus terrestris), and glyphosate's interaction with the gut parasite (Crithidia bombi). We used a fully crossed design measuring bee mortality, parasite intensity and the bacterial composition in the gut microbiome estimated from the relative abundance of 16S rRNA amplicons. We found no impact of either glyphosate, C. bombi, or their combination on any metric, including bacterial composition. This result differs from studies on honeybees, which have consistently found an impact of glyphosate on gut bacterial composition. This is potentially explained by the use of an acute exposure, rather than a chronic exposure, and the difference in test species. Since A. mellifera is used as a model species to represent pollinators more broadly in risk assessment, our results highlight that caution is needed in extrapolating gut microbiome results from A. mellifera to other bee species. [ABSTRACT FROM AUTHOR]

Aims: Since producing more with less is required for increasing agricultural sustainability and reducing its environmental impact, breeding varieties with increased yield stability under reduced fertilizer application is an important goal, particularly in high valued horticultural crops such as tomato (Solanum lycopersicum L.). However, because of the difficulties to conciliate yield and fertilizer use efficiency through breeding, the graft-compatible genetic biodiversity existing in horticultural species offers the possibility to directly approach this objective in high-yielding elite varieties through improving nutrient capture and promoting ecosystem services such as insect pollination. We hypothesized that rootstocks affect pollinator foraging decisions through the nutritional status that impacts yield. Methods: Fifteen genetically diverse experimental rootstocks were grafted to a scion tomato variety and cultivated under optimal and reduced (25% of optimal) P and NPK fertilization in the presence of managed bumblebee pollinators (Bombus terrestris). Results: Up to twofold yield variability between rootstocks was associated with leaf nutrition and photosynthesis of the scion. Interestingly, fertilization regime and the rootstock genotype influenced the pollinator foraging decisions since bumblebees showed feeding preference for plants cultivated under low P, and for the most yielding and nutritious graft combinations under reduced but not under optimal fertilization. Bumblebees can sense plant nutritional status through source-sink relations, as supported by the consistent relationship between pollinator preferences and leaf carbon concentration. Conclusions: This study opens new perspectives for using pollinators as "phenotypers" to select the most resilient plants under suboptimal conditions and/or genotypes that synergistically increase crop productivity by promoting the ecosystem service provided by the insects. [ABSTRACT FROM AUTHOR]

Insect pollination, and in particular pollination by bees, is a highly valued ecosystem service that ensures plant reproduction and the production of high-quality crops. Bee activity is known to be influenced by the weather, and as the global climate continues to change, the flying frequency and foraging behaviour of bees may also change. To maximise the benefits of pollination in a changing world, we must first understand how current weather conditions influence the activity of different bee species. This is of particular interest in a country such as Ireland where inclement weather conditions are nominally sub-optimal for foraging. We observed honeybee (Apis mellifera) and buff-tailed bumblebee (Bombus terrestris) activity across a variety of weather conditions at seven apple orchards to determine how four weather variables (temperature, relative humidity, solar radiation, wind) influenced the flight activity of each species. Each orchard contained three honeybee and three bumblebee colonies, and so we were able to observe a colony of each species concurrently in the same weather conditions. Overall, honeybees were more sensitive to changes in weather than bumblebees and could be more predisposed to future changes in within-day weather conditions. Our results indicate bumblebees could compensate for low honeybee activity in inclement conditions, which supports the theory that pollinator diversity provides resilience. This may be particularly important in management of pollinators in crops that flower in the spring when weather is more variable, and to allow varied responses to global climate change. [ABSTRACT FROM AUTHOR]

Eusocial insect colonies act as a superorganism, which can improve their ability to buffer the negative impact of some anthropogenic stressors. However, this buffering effect can be affected by anthropogenic factors that reduce their colony size. A reduction in colony size is known to negatively affect several parameters like brood maintenance or thermoregulation, but the effects on behaviour and cognition have been largely overlooked. It remains unclear how a sudden change in group size, such as that which might be caused by anthropogenic stressors, affects individual behaviour within a colony. In this study, the bumblebee Bombus terrestris was used to study the effect of social group size on behaviour by comparing the associative learning capabilities of individuals from colonies that were unmanipulated, reduced to a normal size (a colony of 100 workers) or reduced to a critically low but functional size (a colony of 20 workers). The results demonstrated that workers from the different treatments performed equally well in associative learning tasks, which also included no significant differences in the learning capacity of workers that had fully developed after the colony size manipulation. Furthermore, we found that the size of workers had no impact on associative learning ability. The learning abilities of bumblebee workers were thus resilient to the colony reduction they encountered. Our study is a first step towards understanding how eusocial insect cognition can be impacted by drastic reductions in colony size. Significance statement: While anthropogenic stressors can reduce the colony size of eusocial insects, the impact of this reduction is poorly studied, particularly among bumblebees. We hypothesised that colony size reduction would affect the cognitive capacity of worker bumblebees as a result of fewer social interactions or potential undernourishment. Using differential conditioning, we showed that drastic reductions in colony size have no effect on the associative learning capabilities of the bumblebee Bombus terrestris and that this was the same for individuals that were tested just after the colony reduction and individuals that fully developed under the colony size reduction. We also showed that body size did not affect learning capabilities. This resilience could be an efficient buffer against the ongoing impacts of global change. [ABSTRACT FROM AUTHOR]

Bees are important pollinators in wild and agricultural ecosystems, and understanding the factors driving their global declines is key to maintaining these pollination services. Learning, which has been a focus of previous ecotoxicological studies in bees, may play a key role in driving colony fitness. Here we move beyond the standard single-stressor approach to ask how multiple stressors, an agrochemical (sulfoxaflor, a relatively new insecticide) and a parasite (Crithidia bombi, a prevalent gut parasite of bumblebees), impact learning in the bumblebee Bombus terrestris. We developed a modified version of the classic proboscis extension reflex assay to assess the combined effects of acute oral sulfoxaflor exposure and infection by C. bombi on olfactory learning of bumblebee workers. We found no evidence that either sulfoxaflor, C. bombi, or their combination had any significant effect on bumblebee olfactory learning, despite their known negative impacts on other aspects of bumblebee health. This suggests that losses in cognitive ability, as measured here, are unlikely to explain the impacts of sulfoxaflor and its interactions with other stressors on bumblebees. Our novel methodology provides a model system within which to test interactive effects of other key stressors on bee health. [ABSTRACT FROM AUTHOR]

Bees play a vital role as pollinators worldwide and have influenced how flower colour signals have evolved. The Western honey bee, Apis mellifera (Apini), and the Buff-tailed bumble bee, Bombus terrestris (Bombini) are well-studied model species with regard to their sensory physiology and pollination capacity, although currently far less is known about stingless bees (Meliponini) that are common in pantropical regions. We conducted comparative experiments with two highly eusocial bee species, the Western honey bee, A. mellifera, and the Australian stingless bee, Tetragonula carbonaria, to understand their colour preferences considering fine-scaled stimuli specifically designed for testing bee colour vision. We employed stimuli made of pigment powders to allow manipulation of single colour parameters including spectral purity (saturation) or colour intensity (brightness) of a blue colour (hue) for which both species have previously shown innate preferences. Both A. mellifera and T. carbonaria demonstrated a significant preference for spectrally purer colour stimuli, although this preference is more pronounced in honey bees than in stingless bees. When all other colour cues were tightly controlled, honey bees receiving absolute conditioning demonstrated a capacity to learn a high-intensity stimulus significant from chance expectation demonstrating some capacity of plasticity for this dimension of colour perception. However, honey bees failed to learn low-intensity stimuli, and T. carbonaria was insensitive to stimulus intensity as a cue. These comparative findings suggest that there may be some common roots underpinning colour perception in bee pollinators and how they interact with flowers, although species-specific differences do exist. [ABSTRACT FROM AUTHOR]

Bumblebees are important pollinators of wild and agricultural plants but recently have been declining due to various stressors, such as pesticides and diseases. Because of the haplo-diploid sex determination system in hymenopterans, experiments using micro-colonies (small sub colonies without a queen) to identify risks to bumblebee health are limited as they are only able to produce males. Therefore, an experimental protocol for rearing bumblebee larvae in vitro is needed to better understand effects on worker larvae. Here, we aimed to establish a rearing method for larvae of Bombus terrestris for use in risk assessment assays. To confirm the validity of our rearing method, we tested two insecticides used for tomato cultivation, chlorfenapyr and dinotefuran. Bombus terrestris larvae fed with a high nutrient quantity and quality diet increased growth per day. All chlorfenapyr-exposed individuals died within 10 days at 2000-fold dilution, an application dose used for tomatoes. There were significant differences in adult emergence rate among almost all chlorfenapyr treatments. On the other hand, sublethal dinotefuran-exposure did not affect rates of pupation and adult emergence, growth, or larval and pupal periods. Although larvae were smaller than in the natural colony, this rearing method for B. terrestris larvae proved to be effective at evaluating realistic sub-colonies to pesticide exposures. [ABSTRACT FROM AUTHOR]

The way urbanization shapes the intraspecific variation of pollinator functional traits is little understood. However, this topic is relevant for investigating ecosystem services and pollinator health. Here, we studied how urbanization affects the functional traits of workers in two bumblebee species (Bombus terrestris and B. pascuorum) sampled in 37 sites along a gradient of urbanization in North Italy (an area of 1800 km2 including the metropolitan context of Milan and other surrounding capital districts). Namely, we investigated the effect of land use composition, configuration, air temperature, flower resource abundance, and air pollutants on the variation of traits related to flight performance and of stress during insect development (i.e., wing size, wing shape and size fluctuating asymmetry). The functional traits of the two bumblebees responded idiosyncratically to urbanization. Urban temperatures were associated with smaller wing sizes in B. pascuorum and with more accentuated fluctuating asymmetry of wing size in B. terrestris. Moreover, flower abundance correlated with bigger wings in B. terrestris and with less asymmetric wing size in B. pascuorum. Other traits did not vary significantly, and other urban variables played minor effects. These species-specific variation patterns highlight that environmental stressor linked to urbanization negatively impact the traits related to flight performance and development stability of these syntopic bumblebees, with possible consequences on the pollination service they provide. [ABSTRACT FROM AUTHOR]

Living in a social group increases the risks of parasitism, especially in highly-related groups. In homogenous groups, with no reproductive division of labour, the impact of parasitism is unlikely to vary with host identity. Many social systems, however, do exhibit division of reproductive labour, most famously in social insects with their reproductive queens and generally infertile workers. In such systems, the impact of parasitism will differ for each group. Consequently, we predict that susceptibility to parasites will vary to reflect such differential impact. We tested this prediction using a trypanosome-bumble bee system, where Crithidia bombi infects both gynes and workers of Bombus terrestris. We studied both susceptibility to the parasite and relevant measures of the immune function. As predicted, gynes were significantly less susceptible to the parasite than workers, but while gynes and workers expressed different immune profiles, how these link to differential susceptibility remains unclear. In conclusion, our results suggest that differential selection pressures exerted by parasites may produce multiple phenotypes from a single genotype in order to maximise fitness in a social group context. Significance statement: Social insect colonies dominate terrestrial ecology, and as such are targets for parasites. How they defend themselves against such threats is a key question. Here, we show that bumble bee gynes — the reproductive individuals that overwinter and found colonies in this annual social system — are more resistant to a parasite that disproportionately affects reproductive fitness than their sister workers. Differential patterns of susceptibility may help to explain the success of these social insects. [ABSTRACT FROM AUTHOR]

Given that pollinators usually visit flowers for hidden rewards, they need to rely on floral traits that indicate reward status ("honest signals"). However, the relationship between pollination, honest signals, and floral rewards is little documented in natural conditions. The Scotch broom (Cytisus scoparius) is an invasive shrub with polymorphism in the color of its flowers that can be yellow, orange, or red. In three areas dominated by the Scotch broom, we described the abundance of the floral morphs and estimated bumblebee (Bombus terrestris) visitation rate. We examined whether bumblebee visitation to the floral morphs was related to pollen reward. We collected flowers and classified their stamens according to their function: reward or pollen export. Then, we measured anther size and estimated pollen quantity. The yellow morph was more abundant and more visited by bumblebees than the orange and red morphs. The yellow flowers did indeed offer more pollen than the other morphs and this occurred only for rewarding anthers, suggesting that bumblebees could use yellow color as an honest signal to visit the most rewarding flowers. We discuss whether innate and/or learned preferences of bumblebees can explain why the yellow morph is more visited, pollinated, and abundant, while the other morphs are maintained at a lower frequency. This is one of the few field works that shows that variation in intra-specific floral traits is associated with variation in floral reward and pollinator visitation rate, helping to understand the foraging preferences of pollinators and the coexistence of floral morphs in nature. Clinical trials registration: Not applicable. [ABSTRACT FROM AUTHOR]

The bumblebee Bombus terrestris is used worldwide for crop pollination. Despite its positive impact on crop yield, it has become a widespread threat to biodiversity due to its interactions with local bumblebee populations. Commercial subspecies introduced to the Iberian Peninsula since the 1990s without any regulation have colonized the environment, with evidence of naturalization and introgression with the endemic subspecies Bombus terrestris lusitanicus. We have used mitochondrial and nuclear genetic data to describe the current genetic diversity of the Iberian population and to estimate the expansion of commercial bumblebees. Samples from the natural distribution range of the commercial subspecies, the natural intergradation area between the two subspecies and from a period prior to the use of commercial colonies (i.e., before the 1990s) have been used for comparison. Our results show that the mitochondrial haplotype of the commercial breeds has spread throughout the territory, which, together with subtle changes observed in the nuclear genetic diversity of the populations, indicates that hybridization and consequent introgression are occurring in most of the peninsula. It is, therefore, necessary to improve the existing legislation concerning the management and exportation of commercial bumblebees to conserve locally adapted populations. [ABSTRACT FROM AUTHOR]

Social insects have high levels of cooperation and division of labor. In bumble bees this is partly size-based, with larger bees performing tasks outside the nest and smaller bees remaining inside, although bumble bees still display considerable behavioral plasticity. The level of specialization in tasks outside the colony, including foraging, guarding and drifting (entering a foreign colony), is currently unknown for bumble bees. This study aimed to assess division of labor between outside tasks and the degree of specialization in foraging, guarding, and switching colonies in commercially reared bumble bees placed in the field. Nine factory-bought Bombus terrestris colonies were placed on three farms in Sussex, UK, between June and August 2015. Forty workers from each colony were radio-tagged and a reader on the colony entrance recorded the date, time and bee ID as they passed. The length and frequency of foraging trips and guarding behavior were calculated, and drifting recorded. The mean (±SD) length of foraging trips was 45 ± 36 min, and the mean number of foraging trips per day was 7.75 ± 7.71. Low levels of specialization in guarding or foraging behavior were found; however, some bees appeared to guard more frequently than others, and twenty bees were categorized as guards. Five bees appeared to exhibit repeated "stealing" behavior, which may have been a specialist task. The division of labor between tasks was not size-based. It is concluded that commercial bumble bees are flexible in performing outside nest tasks and may have diverse foraging strategies including intra-specific nest robbing. [ABSTRACT FROM AUTHOR]

In recent years, bumblebees have become a prominent insect model organism for a variety of biological disciplines, particularly to investigate learning behaviors as well as visual performance. Understanding these behaviors and their underlying neurobiological principles requires a clear understanding of brain anatomy. Furthermore, to be able to compare neuronal branching patterns across individuals, a common framework is required, which has led to the development of 3D standard brain atlases in most of the neurobiological insect model species. Yet, no bumblebee 3D standard brain atlas has been generated. Here we present a brain atlas for the buff-tailed bumblebee Bombus terrestris using micro-computed tomography (micro-CT) scans as a source for the raw data sets, rather than traditional confocal microscopy, to produce the first ever micro-CT-based insect brain atlas. We illustrate the advantages of the micro-CT technique, namely, identical native resolution in the three cardinal planes and 3D structure being better preserved. Our Bombus terrestris brain atlas consists of 30 neuropils reconstructed from ten individual worker bees, with micro-CT allowing us to segment neuropils completely intact, including the lamina, which is a tissue structure often damaged when dissecting for immunolabeling. Our brain atlas can serve as a platform to facilitate future neuroscience studies in bumblebees and illustrates the advantages of micro-CT for specific applications in insect neuroanatomy. [ABSTRACT FROM AUTHOR]

The ratio of amino acids to carbohydrates (AA:C) that bumble bees consume has been reported to affect their survival. However, it is unknown how dietary AA:C ratio affects other bumble bee fitness traits (e.g., fecundity, condition) and possible trade-offs between them. Moreover, while individual AAs affect phenotype in many species, the effects of AA blend on bumble bee fitness and food intake are unclear. We test how the AA:C ratio that bumble bees (Bombus terrestris) consume affects their condition (abdomen lipid and dry mass), survival following food removal, and ovarian activation. We then compare ovarian activation and food intake in bees fed identical AA:C ratios, but where the blend of AAs in diets differ, i.e., diets contained the same 10 AAs in an equimolar ratio or in the same ratio as in bee collected pollen. We found that AA:C ratio did not significantly affect survival following food removal or ovarian activation; however, high AA intake increased body mass, which is positively correlated with multiple fitness traits in bumble bees. AA blend (i.e., equimolar versus pollen) did not significantly affect overall ovarian activation or consumption of each experimental diet. However, there was an interaction between AA mix and dietary AA:C ratio affecting survival during the feeding experiment, and signs that there may have been weak, interactive effects of AA mix and AA:C ratio on food consumption. These results suggest that the effect of total AA intake on bumble bee phenotype may depend on the blend of individual AAs in experimental diets. We suggest that research exploring how AA blend affects bumble bee performance and dietary intake is warranted, and highlight that comparing research on bee nutrition is complicated by even subtle variation in experimental diet composition. [ABSTRACT FROM AUTHOR]

Bumblebees are important pollinators for many wild plants and crops. To explore gene expression characteristics in depth, stable reference genes will be essential across different developmental stages. In this context, we evaluated the stability of eight common reference genes at ten developmental stages in the bumblebee Bombus terrestris (B. terrestris) with four algorithms and two rank approaches. The results showed that all the primers for the reference genes exhibited high specificity, linear correlation (R2-value 0.9917~0.9978) and a good PCR amplification efficiency (E-value 95.4~108.6%). The combined use of RPL13 and RP49 was suggested to be the most suitable reference genes. Our candidate reference genes will help to improve the precision of the detection of gene expression at different developmental stages of bumblebee species. [ABSTRACT FROM AUTHOR]

Invasive species are one of the main biodiversity loss drivers. Some species can establish and thrive in novel habitats, impacting local communities, as is the case of managed pollinators. In this regard, an invasive species' expansion process over time is critical for its control and management. A good example is the European bumblebee Bombus terrestris, which has rapidly invaded the southern part of South America after being repeatedly introduced in Chile for crop pollination since 1997. We assessed the temporal dynamics of B. terrestris invasion in Argentina and Chile by compiling 562 occurrence points from 2000 to 2019. We used two estimators (minimum convex polygon and 95% fixed kernel) to estimate the increase of the invaded area over time. We found that the area invaded by B. terrestris in the southern part of South America presents a linear increase over time, which was consistent for both estimators. In this scenario, species traits, environmental characteristics, and introduction dynamics facilitate a rapid invasion process that will continue to expand, reaching other South American countries in the near future. As this bumblebee is a super-generalist, it probably will expand across South America, as climate niche modelling predicts, if no actions were taken. [ABSTRACT FROM AUTHOR]

Bombus terrestris is a bumblebee with a wide geographic range, with subspecies showing a variety of local adaptations. Global export of commercially-reared B. terrestris started in the 1980s; the bees are a mixture of subspecies bred for ease of rearing, bivoltinism and large nests. This paper investigated whether the increase in bivoltinism in UK resident B. terrestris audax populations was related to introgression with imported foreign subspecies. Workers were collected from wild populations in London and Bristol, as well as two commercial suppliers. Fourteen microsatellite loci were used to study population structure, hybridisation and introgression. No introgression with commercial B. t. dalmatinus was detected in wild populations. Hence, the increase in winter activity appears unrelated to introgression. [ABSTRACT FROM AUTHOR]

Body temperature changes can modify an insect's ability to fight infections by altering its immune activity. This work evaluated the impact of mild cold and heat thermal stress on the expression of different immune and heat shock genes in the species Bombus terrestris. Additionally, a thermal treatment was repeated under starvation to analyze a possible compromise of immune gene expression in favor of a thermal stress response when energetic resources are limited. Results pointed to a role of Hsc70 and Aha1 genes in tolerance to moderately high temperatures. Expression of the immune genes was not negatively affected by the cold or the heat treatments, and the receptor gene BGRP1 was upregulated with cold, possibly indicating an increase in the cellular immune activity. Under starvation, the effect of the heat and the cold treatments caused a higher upregulation in all genes analyzed, suggesting a synergistic effect of starvation and thermal stress on the activation of heat shock and immune gene expression. [ABSTRACT FROM AUTHOR]

In agricultural landscapes, differences in floral resources provided by crops compared with adjacent habitats promote the spillover of pollinators seeking to fulfil their feeding needs. These foraging patterns play an important role in both crop production and wild plant fitness. However, in classical observational studies, pollinator spillover patterns may be confounded by differences in pollinator phenologies and population sizes. To avoid these confounding effects, we quantified the combined effect of relative floral availability and season on pollinator spillover between strawberry crops and adjacent pinewoods by using commercial bumblebee colonies. We used 60 colonies that were placed in 6 open polytunnel strawberry crops and in 8 pinewoods adjacent to crops in landscapes with either low or high berry (mostly strawberry) crop cover. We repeated the experiment in winter and in spring to account for differences in flower resource availability. While strawberries were in bloom during the entire study period, wild flowering plants were scarce and abundant in winter and in spring, respectively. Spillover in crops was quantified as the percentage of bumblebee individuals that carried pollen from non-berry flowers, while spillover in pinewoods was quantified as the percentage of individuals that carried pollen from berry flowers. Overall, 526 bumblebees were collected. We found them carrying the pollen of 15 plant taxa in winter and 39 in spring, in accordance with seasonal floral availability. In crop colonies, around 30% of bumblebees spilled over into non-berry habitats in winter and 78% in spring, regardless of crop cover in the surrounding landscape. However, in pinewood colonies, we found an interaction effect between season and crop cover: even though on average 34% of bumblebees spilled over into berry crops, in winter it was almost twice in landscapes with high crop cover. Spillover patterns mainly mirrored seasonal changes in floral availability between habitats. [ABSTRACT FROM AUTHOR]

We studied phototaxis, the directional movement relative to light, in the bumble bee Bombus terrestris. We first developed and validated a MATLAB-based system enabling reliable high-resolution tracking of a bee and a measurement of her distance relative to a changing LED light source. Using this system, we found in all our experiments that workers show positive phototaxis. The strength of the phototactic response was influenced by body size but not age, and this effect was significant when the light source was weak. In a separate experiment, foragers showed stronger phototactic response compared with nurses only in one of two trials in which they were larger and tested with weak light intensity. The evidence that phototaxis is associated with size-based division of labor in the bumble bee and with age-related division of labor in the honey bee lends credence to response threshold models implicating the response to light in the organization of division of labor in cavity dwelling social insects. [ABSTRACT FROM AUTHOR]

Recent studies have reported interspecific differences in how bee species respond to various stressors. Evaluating the exposure and responses of different bee species to plant protection products is considered an essential part of their risk assessment. This study was conducted to assess the impacts of thiacloprid-prochloraz mixture on buff-tailed bumblebees (Bombus terrestris) and red mason bees (Osmia bicornis) in a worst-case scenario under semi-field conditions. Bumblebee colonies or solitary bee trap nests were confined in tunnels with flowering oilseed rape. The recommended maximum application rates of 72 g thiacloprid/ha and 675 g prochloraz/ha were applied as a tank mixture during bee flight in full flowering oilseed rape. Several parameters such as flight and foraging activity, population parameters, and exposure level were investigated. Our results show adverse effects of the combination of thiacloprid and prochloraz on the reproductive performance of red mason bees. The number of cocoons produced by O. bicornis was significantly reduced in the treatment compared to the control group. Regarding bumblebees, we found no effects of the thiacloprid-prochloraz mixture on any observed parameters of colony development. The maximum detected concentrations of both active substances three days after application were higher in O. bicornis pollen mass compared to B. terrestris stored pollen. We conclude that this worst-case scenario of thiacloprid-prochloraz exposure poses a high risk to solitary bees and thus the use of such mixture should be restricted. [ABSTRACT FROM AUTHOR]

This study examined shifts over a 35-year period in the phenology of the four most important bumblebee species (Bombus terrestris, B. lapidarius, B. pascuorum and B. hortorum) in Central Europe. The species showed similar temporal trends, significantly advancing components of their main flight period in association with rising temperatures such that, for example, mid-dates of the main flight period advanced by 10–23 days over the study period. Drivers of this change differed between the four species. Trends in, and drivers of, the timing of first queens, first workers and first males were less consistent. Aspects of the phenology of the least common species, B. hortorum, were up to a month earlier than the other species and climatic effects less clear cut. There were some suggestions of differences between species trends. These results stress the importance of considering changes and drivers of change for the Bombus family on a species-specific basis with the need to pay more attention to the life history traits of the study organisms. [ABSTRACT FROM AUTHOR]

Bumblebees are important pollinating insects, but many species have suffered declines over the last century. Long-tongued bumblebees have been identified as particularly at risk, partly due to their more selective diet. Attempts to study these species in captivity have been impeded by stress-induced behaviours which cause queens to kill or abandon their brood. Here, we attempt to further develop techniques, using queen pairing and Bombus terrestris cocoons, to successfully rear two common long-tongued bumblebee species (B. pascuorum and B. hortorum) in captivity. Approximately half of queens laid eggs and 29% produced workers. Although challenges remain, there is a great deal to be gained from optimising the captive rearing of these species. [ABSTRACT FROM AUTHOR]

The invasion of alien pollinators may disturb the local pollination network between plants and native pollinators. However, alien pollinators may be able to coexist with native pollinators if spatiotemporal partitioning of floral resources is possible at the landscape scale. We compared the seasonal patterns of floral choice and habitat use across local habitats (natural coastal grassland, semi-natural grassland and arable field) between alien Bombus terrestris and two major native species, Bombus hypocrita and Bombus deuteronymus, in a coastal area of northern Japan. Although two native species with different morphological characteristics showed specific floral use patterns, that of B. terrestris overlapped with both native species. Short-tongued B. terrestris and B. hypocrita frequently visited Rosa rugosa and Calystegia soldanella flowers that have shallow or open corollas in the coastal grassland. B. terrestris also visited legume flowers with a long-tubed shape at high frequency in both semi-natural and coastal grasslands, as did long-tongued B. deuteronymus despite the difference in morphology. B. terrestris was observed in every habitat type based on the abundance of floral resources. B. terrestris had wide foraging area and changed target flowers flexibly at the landscape scale according to the seasonal changes in floral resources. Because of the spatiotemporally diverse foraging patterns between B. terrestris and native bumblebees, competition for floral resources may not be intensive at the landscape scale owing to the diverse plant communities mixed with natural and non-natural habitats. [ABSTRACT FROM AUTHOR]

Bumblebees Bombus terrestris are good at learning to distinguish between patterned flowers. They can differentiate between flowers that differ only in their patterning of scent, surface texture, temperature, or electrostatic charge, in addition to visual patterns. As recently shown, bumblebees trained to discriminate between nonvisual scent patterns can transfer this learning to visually patterned flowers that show similar spatial patterning to the learnt scent patterns. Bumblebees can, therefore, transfer learnt patterns between different sensory modalities, without needing to relearn them. We used differential conditioning techniques to explore whether cross-modal transfer of learnt patterns also occurred between visual and temperature patterns. Bumblebees that successfully learnt to distinguish rewarding and unrewarding temperature patterns did not show any preferences for the corresponding unlearnt visual pattern. Similarly, bumblebees that learnt visual patterns did not transfer these to temperature patterns, suggesting that they are unable to transfer learning of temperature and visual patterns. We discuss how cross-modality pattern learning may be limited to modalities that have potentially strong neurological links, such as the previously demonstrated transfer between scent and visual patterns. [ABSTRACT FROM AUTHOR]

Context: Bumblebees are important pollinators for agricultural crops and wild plants. However, agricultural intensification and loss of semi-natural habitat may have adverse effects on colony performance. While mass-flowering crops may serve as food sources, landscapes dominated by intensive agriculture may be a poor bumblebee habitat compared to landscapes dominated by semi-natural habitat, such as forests and species-rich grasslands. Objectives: We investigated the effect of different land use classes on bumblebee colony performance at two periods of time during the flying season. We expected mass-flowering crops to have a positive influence on colony performance during flowering. After mass-flowering, presence of semi-natural habitat was hypothesized to be essential for colony development. Methods: We placed artificial colonies of Bombus terrestris in 28 apple and pear orchards during and after mass-flowering. We measured colony growth and took pollen samples from foraging workers. Causal relationships between land-use, pollen quality, amount of pollen collected and colony performance were analyzed. Results: Contrary to our expectations, we found a negative correlation between orchard cover and amount of pollen collected in spring, during mass-flowering, resulting in a lower colony performance. During mass-flowering, colony performance and pollen diversity were positively correlated with cover of open semi-natural habitat, while after mass-flowering, colony performance was positively correlated with forest cover. After mass-flowering, pollen sources were more diverse and colony growth was related to land use classes at a larger spatial scale, indicating that bumblebees foraged further. We did not find a correlation between the performances of colonies at one specific site that were placed during and after mass-flowering. Conclusions: Landscapes dominated by mass-flowering orchards represent inferior habitats for bumblebees during flowering. Heterogenous landscapes, containing a mix of various types of semi-natural habitat increase the habitat quality for bumblebees. We advise to maintain these diverse patches of semi-natural habitat, as mass-flowering crops alone are insufficient to support bumblebee colonies. [ABSTRACT FROM AUTHOR]

Recent anthropogenic fragmentation has led to population differentiation threatening viability of many species, including species specialized on mountainous ecosystems. Bombus ephippiatus, a widespread species mostly found in mountains in the Neotropics, seems to use the highlands as island, and deforested lowland areas may represent barriers to their dispersal, leading to isolation and potentially loss of genetic diversity. Yet, lack of knowledge of its population structure does not allow adequate management and conservation. To fill this knowledge gap, we assessed the population structure and inferred dispersion of B. ephippiatus in two mountain-ranges in Guatemala (Volcanic Chain and Sierra de las Minas). This region is characterized by high topographic variation and considerable deforestation strain. We analyzed the effects of elevation and land-use on genetic differentiation of B. ephippiatus populations and inferred its demography in the region. Our results suggest that B. ephippiatus is able to disperse long distances across most landscape types, reflected by its high genetic diversity, high effective population size, considerable gene flow, low population differentiation, as well as the lack of isolation by distance. Hence, B. ephippiatus may be a resilient species for the provision of pollination services. However, we detected a subtle divergence of B. ephippiatus into two clusters, of which Sierra de las Minas has been identified as a regional hotspot of genetic and species endemism. Yet, differentiation is very recent and hence likely caused by lowland deforestation. The combined effects of current forest cover and elevation partially explain the observed subtle patterns of differentiation suggesting that the maintenance of suitable habitat is crucial to ensure population connectivity of this keystone pollinator. [ABSTRACT FROM AUTHOR]

Many insects rely on vision to find food, to return to their nest and to carefully control their flight between these two locations. The amount of information available to support these tasks is, in part, dictated by the spatial resolution and contrast sensitivity of their visual systems. Here, we investigate the absolute limits of these visual properties for visually guided position and speed control in Bombus terrestris. Our results indicate that the limit of spatial vision in the translational motion detection system of B. terrestris lies at 0.21 cycles deg with a peak contrast sensitivity of at least 33. In the perspective of earlier findings, these results indicate that bumblebees have higher contrast sensitivity in the motion detection system underlying position control than in their object discrimination system. This suggests that bumblebees, and most likely also other insects, have different visual thresholds depending on the behavioral context. [ABSTRACT FROM AUTHOR]

The bumblebee ( Bombus terrestris) has become a common model animal in the study of various aspects of vision and visually guided behavior. Although the bumblebee visual system has been studied to some extent, little is known about the functional role of the first visual neuropil, the lamina. In this work, we provide an anatomical and electrophysiological description of the first-order visual interneurons, lamina monopolar cells (LMCs), of the bumblebee. Using intracellular recording coupled with dye injection, we found that bumblebee LMCs morphologically resemble those found in the honeybee, although only the LMC type L1 cells could be morphologically matched directly between the species. LMCs could also be classified on the basis of their light response properties as spiking or non-spiking. We also show that some bumblebee LMCs can produce spikes during responses to stimulation with naturalistic light contrasts, a property unusual for these neurons. [ABSTRACT FROM AUTHOR]

Slow bee paralysis virus (SBPV)-previously considered an obligate honeybee disease-is now known to be prevalent in bumblebee species. SBPV is highly virulent in honeybees in association with Varroa mites, but has been considered relatively benign otherwise. However, condition-dependent pathogens can appear asymptomatic under good, resource abundant conditions, and negative impacts on host fitness may only become apparent when under stressful or resource-limited conditions. We tested whether SBPV expresses condition-dependent virulence in its bumblebee host, Bombus terrestris, by orally inoculating bees with SBPV and recording longevity under satiated and starvation conditions. SBPV infection resulted in significant virulence under starvation conditions, with infected bees 1.6 times more likely to die at any given time point (a median of 2.3 h earlier than uninfected bees), whereas there was no effect under satiated conditions. This demonstrates clear condition-dependent virulence for SBPV in B. terrestris. Infections that appear asymptomatic in non-stressful laboratory assays may nevertheless have significant impacts under natural conditions in the wild. For multi-host pathogens such as SBPV, the use of sentinel host species in laboratory assays may further lead to the underestimation of pathogen impacts on other species in nature. In this case the impact of 'honeybee viruses' on wild pollinators may be underestimated, with detrimental effects on conservation and food security. Our results highlight the importance of multiple assays and multiple host species when testing for virulence, in order for laboratory studies to accurately inform conservation policy and mitigate disease impacts in wild pollinators. [ABSTRACT FROM AUTHOR]

Flying insects frequently navigate through environments of different complexity. In this study, buff-tailed bumblebees ( Bombus terrestris L.) were trained to fly along tunnels of different widths, from 60 to 240 cm. In tunnel widths of 60 and 120 cm, bumblebees control their lateral position by balancing the magnitude of translational optic flow experienced in the lateral visual field of each eye. In wider tunnels, bumblebees use translational optic flow cues in the ventral visual field to control their lateral position and to steer along straight tracks. Our results also suggest that bumblebees prefer to fly over surfaces that provide strong ventral optic flow cues, rather than over featureless ones. Together, these strategies allow bumblebees to minimize the risk of collision and to maintain relatively straight flight paths in a broad range of environments. [ABSTRACT FROM AUTHOR]

In this study, we investigated if oral administration of lactic acid bacteria could increase the colony performance of reared bumblebees. We found that a continuous administration of Lactobacillus kunkeei LMG 18925 and Lactobacillus crispatus LMG 9479 could partly compensate the effects of low nutritional pollen, but a permanent colonization in the gut was not detected with Illumina MiSeq 16S rRNA sequencing. Contrary to expectations, a single administration of L. kunkeei LMG 18925 to a high nutritional pollen led to a lower total drone mass. A Bombus-specific strain, Bifidobacterium actinocoloniiforme R-53049, showed potential to colonize the gut permanently after three administrations. Our study represents a first screening for the potential use of probiotic strains in bumblebees. We conclude that both diet and host specificity of bacteria might have an effect on colony performance of indoor-reared bumblebees and play a role in the gut colonization success. [ABSTRACT FROM AUTHOR]

In most bumblebee species, queens mate with only one male, but males can mate more than once. However, the effects of male remating on colony foundation success of queens remain poorly understood. To test for effects of male remating, we assigned Bombus terrestris queens randomly to one of two groups: virgin males or non-virgin males that have mated once before. When males were allowed to mate for a second time with virgin queens, 58.57 % successfully remated. There was no significant difference in queen hibernation survival between groups, but queens mated with non-virgin males were more successful in founding a colony, and produced more workers and males than queens mated with virgin males. The mean copulation duration for the second mating of males (48.02 ± 14.36 min) was substantially longer than for the first mating (32.44 ± 8.74 min). Results also showed that male weight influenced both male remating success and copulation duration. [ABSTRACT FROM AUTHOR]

The aim of this study was to investigate the effects of Elado-dressed winter oilseed rape (OSR, 10 g clothianidin & 2 g beta-cyfluthrin/kg seed) on the development, reproduction and behaviour of large earth bumble bees ( Bombus terrestris) as part of a large-scale monitoring field study in Northern Germany, where OSR is usually cultivated at 25-33 % of the arable land. Both reference and test sites comprised 65 km in which no other crops attractive to pollinating insects were present. Six study locations were selected per site and 10 bumble bee hives were placed at each location. At each site, three locations were directly adjacent to OSR fields and three locations were situated 400 m distant from the nearest OSR field. The development of colonies was monitored from the beginning of OSR flowering in April until June 2014. Pollen from returning foragers was analysed for its composition. An average of 44 % of OSR pollen was found in pollen loads of bumble bees indicating that OSR was a major resource for the colonies. At the end of OSR flowering, hives were transferred to a nature reserve until the end of the study. Colony development in terms of hive weight and the number of workers showed a typical course with no statistically significant differences between the sites. Reproductive output was comparatively high and not negatively affected by the exposure to treated OSR. In summary, Elado-dressed OSR did not cause any detrimental effects on the development or reproduction of bumble bee colonies. [ABSTRACT FROM AUTHOR]

Conservation action for species of concern requires that 'designatable units' (e.g., species, subspecies, geographic races, genetically distinct forms) are clearly defined, or that the species complex is treated as a whole. Several species of bumble bee are currently threatened, and some of these have cryptic colouration (resembling other species), or form complexes that vary considerably in colour patterning. Here we address the taxonomy and distribution of Bombus occidentalis Greene and B. terricola Kirby, both of which are currently of conservation concern in North America. Bombus occidentalis includes two apparently monophyletic groups of COI barcode haplotypes (recently considered as subspecies) with ranges mostly separated by that of their sister species, B. terricola. The southern B. o. occidentalis ranges throughout the western United States and into western Canada from southern Saskatchewan and Alberta, and throughout British Columbia north to ca. 55°N; the northern B. o. mckayi Ashmead, is restricted to north of this in British Columbia, westernmost Northwest Territories, Yukon Territory and Alaska. Bombus o. mckayi exists, as far as is known, only with a 'banded' colour pattern. By contrast, B. o. occidentalis occurs in both banded and non-banded colour patterns, although the southern banded colour pattern is geographically isolated from the northern subspecies. Bombus o. occidentalis has declined throughout its range, perhaps due in part to exposure to novel parasites. Despite having similar levels of parasitism (ca. 40 %) as the southern subspecies, B. o. mckayi appears to have stable populations at present. There is therefore compelling evidence that the two subspecies should be distinguished for conservation and management purposes. We present the evidence for their distinction and provide tools for subspecies recognition. [ABSTRACT FROM AUTHOR]

Bumblebees demonstrate an extensive capacity for learning complex motor skills to maximise exploitation of floral rewards. This ability is well studied in nectar collection but its role in pollen foraging is less well understood. Floral sonication is used by bees to extract pollen from some plant species with anthers which must be vibrated (buzzed) to release pollen. Pollen removal is determined by sonication characteristics including frequency and amplitude, and thus the ability to optimise sonication should allow bees to maximise the pollen collection. We investigated the ability of the buff-tailed bumblebee ( Bombus terrestris) to modify the frequency and amplitude of their buzzes with increasing experience manipulating flowers of the buzz-pollinated plant Solanum rostratum. We analysed flight and feeding vibrations generated by naïve workers across feeding bouts. Feeding buzzes were of a higher frequency and a lower amplitude than flight buzzes. Both flight and feeding buzzes had reduced amplitudes with increasing number of foraging trips. However, the frequency of their feeding buzzes was reduced significantly more than their flight buzzes as bumblebee workers gained experience manipulating flowers. These results suggest that bumblebees are able to modify the characteristics of their buzzes with experience manipulating buzz-pollinated flowers. We discuss our findings in the context of bumblebee learning, and the current understanding of the optimal sonication characteristics for releasing pollen in buzz-pollinated species. Our results present a tantalising insight into the potential role of learning in floral sonication, paving the way for future research in this area. [ABSTRACT FROM AUTHOR]

Kaolin is an inert material with a broad range of applications, e.g. as an insecticide and as a filling substance in the formulation of biopesticides. Hence, bees that dispense biopesticides to the field in the context of entomovectoring are exposed to elevated risks because of side-effects of those products. Here, we investigated with use of bumble bee workers of Bombus terrestris L. the lethal and sublethal effects of (i) pure kaolin, (ii) the biofungicide Prestop-Mix containing the parasitic fungus Gliocladium catenulatum and kaolin and (iii) the bioinsecticide BotaniGard containing the entomopathogenic fungus Beauveria bassiana and compared with wheat flour powder that we considered safe for bumble bees. As the most important result, treatment with kaolin increased the cuticular water loss and reduced the survival of treated bumble bees, while Prestop-Mix had no negative effect on longevity. BotaniGard caused mortality in the bumble bees because of the entomopathogenic spores it contained. In conclusion, our data indicated that substances used as 'inert materials' and in biocontrol agents which are used in IPM and organic farming systems may bring higher risks to bumble bees used in entomovector technology. [ABSTRACT FROM AUTHOR]

Flying animals are capable of navigating through environments of different complexity with high precision. To control their flight when negotiating narrow tunnels, bees and birds use the magnitude of apparent image motion (known as optic flow) generated by the walls. In their natural habitat, however, these animals would encounter both cluttered and open environments. Here, we investigate how large changes in the proximity of nearby surfaces affect optic flow-based flight control strategies. We trained bumblebees to fly along a flight and recorded how the distance between the walls-from 60 cm to 240 cm-affected their flight control. Our results reveal that, as tunnel width increases, both lateral position and ground speed become increasingly variable. We also find that optic flow information from the ground has an increasing influence on flight control, suggesting that bumblebees measure optic flow flexibly over a large lateral and ventral field of view, depending on where the highest magnitude of optic flow occurs. A consequence of this strategy is that, when flying in narrow spaces, bumblebees use optic flow information from the nearby obstacles to control flight, while in more open spaces they rely primarily on optic flow cues from the ground. [ABSTRACT FROM AUTHOR]

Bumblebees of Bombus terrestris are indispensible pollinators for ecosystems and for various agricultural crops. Unfortunately, bumblebees are challenged by various stress factors including insecticide applications. Today sublethal effects of various insecticides need to be thoroughly investigated to allow their combined use with pollinators and other beneficial organisms. In this study, we used lambda-cyhalothrin as a model pyrethroid insecticide and investigated lethal and sublethal effects by different dilutions, ranging from 1/10 to 1/100 of its maximum field recommended dosage (MFRC, 37.5 ppm), with the use of a chronic toxicity tests in the laboratory and in flight cages in the greenhouse. In the laboratory, small microcolonies with five bumblebee workers with one being pseudo-queen were used, while in the greenhouse we used queen-right mini-hives where the bumblebees need to fly for pyrethroid-contaminated food. We observed strong sublethal effects in the laboratory with treatments of 1/10 and 1/20 of the MFRC: the nest reproduction was reduced by 49 and 32 %, respectively, and the sugar water consumption by 36 %. With free-flying bumblebees, the toxic effects at 1/10 of the MFRC were more pronounced. A mortality of 88 ± 8 % was observed after only 2 weeks, being twice the mortality in the laboratory microcolonies test (43 ± 11 %). Besides, it should be mentioned that in the greenhouse experiment all queens were dead and most of the workers showed signs of incoordination and convulsion and gradually became apathetic. In conclusion, our results demonstrated the pyrethroid lambda-cyhalothrin with a range of lethal and sublethal effects, both crucial for the development and survival of the B. terrestris colonies. Moreover, this study supports the demand to test insecticide compounds on their safety, especially when the bees have to perform complex tasks such as foraging for their food. [ABSTRACT FROM AUTHOR]

Many species display local variations in pre-mating signals and in mating preferences. This may lead to discrimination against potential foreign mates that may ultimately lead to reproductive isolation. However, the extent to which population differentiation in mating cues affects the species recognition has received little empirical support. Here, we investigate the consequence of geographic differentiation in male reproductive traits on female preferences to these traits in Bombus terrestris. We characterise (1) the geographic differentiation in male cephalic labial gland secretions (CLGS), a key trait for mate attraction, and (2) the preference of virgin females to the CLGS of different subspecies. Our results show geographic CLGS differences parallel with divergences in female preferences for these secretions. This geographic CLGS differentiation in males, along with female preference for sympatric males, could lead to or reflect a pre-mating isolation among subspecies. [ABSTRACT FROM AUTHOR]

Bumblebee queens require a continuous supply of flowering food plants from early spring for the successful development of annual colonies. Early in spring, Corydalis cava provides essential nectar and pollen resources and a choice of flower colour. In this paper, we examine flower colour choice (purple or white) in C. cava and verify the hypothesis that bumblebee queens differ in their choice of flower colour. A total of 10,615 observations of flower visits were made in spring 2011 and spring 2014 near Poznań, western Poland. Our results suggest that Bombus lucorum/cryptarum used purple flowers less, while Bombus terrestris used purple flowers more and Bombus hortorum showed no preference. Therefore, the colour morphs of C. cava are probably co-evolutionary adaptations to the development of another part of the insect community which has different colour preferences. [ABSTRACT FROM AUTHOR]