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Nectar-foraging bees change their use of floral resources as plant species appear in the environment and disappear over their lifetimes. The new flowers used may involve different cues and different nectar extraction tactics. Although bumble bees can adapt to changes in floral cues and required tactics, little is known about whether bees prioritize switching tactics or floral cues when deciding which plant species to switch to. In a laboratory assay, we forced common eastern bumble bee, Bombus impatiens , workers either to switch the handling tactic they were using or to continue using the tactic but to switch the colour of artificial flowers foraged on. We examined whether bees' tendency to change their tactics was influenced by how similar in colour novel flowers were to familiar ones. We conducted a 2 × 2 factorial experiment using artificial flowers, manipulating the handling tactic that bees were initially trained to use (legitimate visit or nectar robbing) and the similarity between novel and trained colours (similar or distinct). We found that under most conditions bees preferred to switch flower colours and retain handling tactics. However, when given experience with legitimate visits and when novel flowers were markedly different in colour from those they had experienced previously, bees tended to switch tactics while continuing to forage on flowers of the same colour. These findings suggest that the similarity in colour of a new floral resource to the currently exploited resource and the flower-handling tactic used by bees both play an important role in decision making by foraging bumble bees. • We examined bumble bees' switching decisions to novel types of flowers. • Bees were trained to legitimately visit or rob blue flowers to obtain reward. • Novel (dark blue or yellow) flower rewards required the trained tactic. • Bees switched to dark blue flowers using the trained tactic. • Bees switched to robbing blue flowers when the novel flower was yellow. [ABSTRACT FROM AUTHOR]

Bumble bees are the primary commercial pollinators for greenhouse‐grown crops and managing them requires insight into the Bombus life cycle. However, some aspects of the life cycle are not fully understood. Once mated, adult Bombus gynes can be stored for a short term at a constant low temperature (CLT). However, survival rates for some species are low during CLT, particularly when storage periods exceed a month. To address this, we tested the hypothesis that Bombus gynes stored in a fluctuating thermal regime (FTR) have increased survival compared to gynes stored in CLT. Specifically, the FTR had a baseline temperature of 2°C, with a daily fluctuation to 20°C relative to the CLT of 2°C with no warming pulse. We tested the effects of FTR on gyne survival and post‐cold storage lipid content in three species used for commercial pollination in North America: B. vosnesenskii, B. huntii and B. impatiens. We further assessed how additional factors influence cold storage survival, including gyne body weight, body size and colony age. Survival responses varied among species. Bombus vosnesenskii and B. huntii had a lower survival rate in FTR compared to CLT, but B. impatiens gynes had a significantly higher survival rate in FTR. We did not observe significant effects of FTR on lipid stores for any species, but survival increased with an increasing initial weight prior to cold storage. Moreover, gynes produced later in the colony life cycle had lower body weight and decreased survival compared to early emerging gynes. Our study provides insight into how cold storage conditions interact with colony and gyne characteristics to influence captive overwintering survival. Our findings have economic applications for improving commercial rearing and efforts for conservation rearing of at‐risk Bombus species. [ABSTRACT FROM AUTHOR]

Pollinators face many stressors, including reduced floral diversity. A low-diversity diet can impair organisms’ ability to cope with additional stressors, such as pathogens, by altering the gut microbiome and/or immune function, but these effects are understudied for most pollinators. We investigated the impact of pollen diet diversity on two ecologically and economically important generalist pollinators, the social bumble bee (Bombus impatiens) and the solitary alfalfa leafcutter bee (Megachile rotundata). We experimentally tested the effect of one-, two-, or three-species pollen diets on gut bacterial communities in both species, and the melanization immune response in B. impatiens. Pollen diets included dandelion (Taraxacum officinale), staghorn sumac (Rhus typhina), and hawthorn (Crataegus sp.) alone, each pair-wise combination, or a mix of all three species. We fed bees their diet for 7 days and then dissected out guts and sequenced 16S rRNA gene amplicons to characterize gut bacterial communities. To assess melanization in B. impatiens, we inserted microfilament implants into the bee abdomen and measured melanin deposition on the implant. We found that pollen diet did not influence gut bacterial communities in M. rotundata. In B. impatiens, pollen diet composition, but not diversity, affected gut bacterial richness in older, but not newly-emerged bees. Pollen diet did not affect the melanization response in B. impatiens. Our results suggest that even a monofloral, low-quality pollen diet such as dandelion can support diverse gut bacterial communities in captive-reared adults of these bee species. These findings shed light on the effects of reduced diet diversity on bee health. [ABSTRACT FROM AUTHOR]

Community diversity can reduce the prevalence and spread of disease, but certain species may play a disproportionate role in diluting or amplifying pathogens. Flowers act as both sources of nutrition and sites of pathogen transmission, but the effects of specific plant species in shaping bee disease dynamics are not well understood. We evaluated whether plantings of sunflower (Helianthus annuus), whose pollen reduces infection by some pathogens when fed to bees in captivity, lowered pathogen levels and increased reproduction in free-foraging bumblebee colonies (Bombus impatiens). Sunflower abundance reduced the prevalence of a common gut pathogen, Crithidia bombi, and reduced infection intensity, with an order of magnitude lower infection intensity at high sunflower sites compared with sites with little to no sunflower. Sunflower abundance was also positively associated with greater queen production in colonies. Sunflower did not affect prevalence of other detected pathogens. This work demonstrates that a single plant species can drive disease dynamics in foraging B. impatiens, and that sunflower plantings can be used as a tool for mitigating a prevalent pathogen while also increasing reproduction of an agriculturally important bee species.

The presence of secondary metabolites in flower nectar can mediate interactions between plants, pollinators, herbivores, and microbes. Milkweeds range in concentrations of cardenolides in flower nectar from ~ 1 to 100 ng μl–1. Using three different behavioral assays with bumblebees Bombus impatiens, we examined the impacts of the commercially available cardenolide ouabain at the range of concentrations at which cardenolides naturally occur in milkweeds. We show that after four days of exposure bees in consumption assays drank more of a nectar solution with a low ouabain concentration of 10 ng μl–1 than the control sucrose nectar, and over the course of the experiment bees consumed less of the 100 ng μl–1 ouabain solution than the control. Bee activity levels in Petri dish arena assays were not impacted by ouabain consumption, even at the highest concentrations; however, in free‐flying choice assays, bees preferentially visited artificial flowers containing 10 ng μl–1 ouabain more than flowers with sucrose control, or flowers with 100 ng μl–1 of ouabain. We therefore conclude that cardenolides may provide plants advantages to pollination at the low end of the naturally occurring range of concentrations for cardenolides, but may be costly to plants at the high end of the range. This research highlights that secondary metabolites in nectar may be under selection to be maintained at low concentrations, and species with high concentrations of secondary metabolites may be under conflicting selection pressures to maintain high circulating levels to deter herbivores at the cost of reduced pollination. [ABSTRACT FROM AUTHOR]

Pollinators are threatened by diverse stressors, including microbial pathogens such as Crithidia bombi. Consuming sunflower pollen dramatically reduces C. bombi infection in the bumble bee Bombus impatiens, but the mechanism behind this medicinal effect is unclear. We asked whether diet mediates resistance to C. bombi through changes in the gut microbiome. We hypothesized that sunflower pollen changes the gut microbiome, which in turn reduces Crithidia infection. To test this, we performed a gut transplant experiment. We fed donor bees either a sunflower pollen treatment or buckwheat pollen as a control treatment and then inoculated recipient bees with homogenized guts from either sunflower-fed or buckwheat-fed donor bees. All recipient bees were then fed a wildflower pollen diet. Two days after the transplant, we infected recipients with C. bombi, and 2 days later, we provided another donor gut transplant. To quantify infection, we performed both fecal screens and dissections of the recipient bees. We found no significant differences in C. bombi infection intensity or presence between bees that received sunflower-fed microbiomes versus buckwheat-fed microbiomes. This suggests that sunflower pollen's effects on pathogen resistance are not mediated by gut microbiota. [ABSTRACT FROM AUTHOR]

Bumble bees are globally important pollinators, contributing hundreds of millions of dollars annually in crop pollination services. Several species are in decline, making it paramount to understand how pathogens and nutrition shape bee health. Previous work has shown that consuming sunflower pollen (Helianthus annuus) dramatically reduces infection by the trypanosomatid gut pathogen, Crithidia bombi , in the common eastern bumble bee (Bombus impatiens). Sunflower pollen may therefore be useful as a dietary supplement for reducing this pathogen in managed bumble bee colonies. Here, we assessed the efficacy of freezer-stored sunflower pollen that was collected in different years and locations for reducing pathogen infection. We tested sunflower pollen that was 1, 3, 4, or 5 yr old and from sunflowers grown in the United States or China against a control of 1-yr-old buckwheat pollen from China, since buckwheat pollen results in high infection. We hypothesized that older pollen would have weaker medicinal effects due to degradation of pollen quality. We found that all sunflower pollen treatments significantly decreased Crithidia infection compared to controls. These results suggest that sunflower pollen can be freezer-stored for up to 5 yr and sourced from a wide range of geographic areas and still maintain its medicinal effects against Crithidia in the common eastern bumble bee. This is helpful information for stakeholders who might administer sunflower pollen as a dietary supplement to manage Crithidia in commercial bumble bee colonies. [ABSTRACT FROM AUTHOR]

Parents that provide parental care must balance the conservation of resources with the benefits of expenditure in support of existing offspring. There is added complexity in social systems in which siblings also contribute to brood care, wherein dynamic social interactions exist, and brood care can influence group fitness. Most bumble bees are annually eusocial insects in which queens undergo a stark transition from living a solitary, to subsocial, to eusocial lifestyle over the course of several weeks as they initiate new nests. We used detailed behavioural analyses of queen bumble bee (Bombus impatiens) parental care behaviour to examine how performance of five parental care-related tasks differs when queens are living in either subsocial conditions (with brood but no adult workers) or as part of a eusocial group with either three or five workers. These three groups reflect the ontogeny of sociality across the early nesting stage. We show for the first time that queen bumble bee parental care behaviour is tightly regulated by the number of helpers in the nest. With three workers in the nest, queens exhibited ∼60% reduction in brood feeding and ∼30% reduction in incubation but no reduction in food collection. However, the addition of just two additional workers resulted in nearly complete cessation of pollen and nectar collection behaviours, whereas in-nest parental care behaviours persisted. We propose that this sequence of behavioural changes might reflect queen avoidance of threats such as predators, parasites or harsh weather. Queens may cease performing tasks outside the nest early in colony development, but only when a threshold number of helpers in the nest is met. More broadly, our results suggest that bumble bee queen parental care behaviour is heavily impacted by the number of helpers in the nest, who may alleviate some of the constraints queens face during nest initiation. • Bumble bee queens dynamically modulated parental care effort in response to helpers. • In-nest task performance was slightly reduced in the presence of three workers. • Out-of-nest task performance was dramatically reduced in the presence of five workers. [ABSTRACT FROM AUTHOR]

Environmental stressors frequently have sublethal consequences for animals, often affecting the mean of phenotypic traits in populations. However, effects on inter‐individual variability are poorly understood. Since phenotypic variability is the basis for adaptation, any change due to stressors may have important implications for population resilience.Here, we explored this possibility in bees by analysing raw datasets from 23 studies (5618 bees) in which individuals were first exposed to stressors and then tested for cognitive tasks.While all types of stressors decreased the mean cognitive performance of bees, they increased cognitive variability. Focusing on 14 pesticide studies, we found that the mode of exposure to stressors and the dose were critical. Mean cognitive performance was more affected by a chronic exposure than by an acute exposure. Yet, cognitive variability increased with increasing doses following both exposure durations.Policy implications. Current guidelines for the authorization of plant protection products on the European market prioritize acute over chronic toxicity assessments on non‐target organisms. By overlooking the consequences of a chronic exposure, regulatory authorities may register new products or doses that are harmful to bee populations. Our findings call for more research on stress‐induced phenotypic variation and its incorporation into policy guidelines to help identify levels and modes of exposure animals can cope with. [ABSTRACT FROM AUTHOR]

Floral scents shape plant–pollinator interactions. Although populations of the same species can vary in their floral scent, little is known about how this variation affects pollinator visitation. In this study, we compare the scents emitted by buzz-pollinated Solanum rostratum (Solanaceae) in two areas of its distribution (Mexico and USA) and investigate how these differences in scent affect pollinator preferences and attraction. We determined the variation of floral volatile compounds using hexane extraction followed by gas chromatography coupled with spectrometry. We also performed a field cage multiple-choice bioassay and a Y-tube behavioural bioassay using Bombus impatiens. We recorded 13 volatile compounds in floral extracts for plants from both ranges that varied qualitative and semi-quantitatively among populations. We found that in the field cage experiment, bumble bees visited plants from the US populations more frequently than plants from Mexican populations. However, bees showed no difference in preference between extracts from Mexican or US flowers. We conclude that although bees show differential visitation to whole plants of different regions, variation in floral extract alone does not translate into differences in preference by B. impatiens. The potential effects of variation in floral scent on the other native bee pollinators remain to be assessed. [ABSTRACT FROM AUTHOR]

A key question in pollination biology is that of how pollinators identify and choose foraging patches. Several approaches have been employed for this, including field studies and large greenhouse flight chambers. Most methods used to date are limited, though, by reliance on a combination of artificial flowers, large spatial scales, or lack of spatially distinct floral patches. To address this issue, we designed and tested a y-maze flight arena and tested it using the bumblebee Bombus impatiens and canola plants. Our results indicate that the system is not biased by environmental conditions, or by an innate "handedness" of Bombus impatiens. We found that bees made all the expected patch choices when presented with soil, plants without flowers, or plants with flowers. This new method is important and useful as it allows researchers to ask questions of both plant health and insect behavior and the chamber system is modular allowing for simple changes to the setup to focus on different questions. Highlights: • Y-maze flight arena was designed to evaluate foraging behavior on intact plants. • No evidence of side preference in individual bees. • Behaviors observed in the y-maze appear to correspond to behaviors observed in other settings. [ABSTRACT FROM AUTHOR]

Due to a lack of knowledge on the pollination requirements of kiwifruit cultivars grown within the United States, farmers simultaneously implement multiple pollination methods, like the rental of managed bee species or artificial pollination to achieve high fruit yields. However, implementing multiple pollination methods is costly and possibly an inefficient use of resources. We assessed the contribution of two managed bees (Apis mellifera and Bombus impatiens) to the pollination of kiwifruit by i) determining the relative abundance of kiwifruit pollen collected by foragers of each bee species, and ii) comparing fruit set and fruit quality among insect and artificially pollinated flowers through an insect exclusion experiment. A significant difference was observed between the mean relative abundance of kiwifruit pollen carried in the corbicula of A. mellifera and B. impatiens, with B. impatiens carrying on average 46% more kiwifruit pollen than A. mellifera. Artificially pollinated kiwifruit flowers set significantly greater numbers of fruit per flower at four weeks post-bloom and at harvest compared to insect pollination, wind pollination, and pollen exclusion treatment. Artificial pollination produced fruits of greater weight, size, and seed number compared to insect-pollinated flowers, and few fruits were produced in the pollen exclusion and wind pollination treatments. Kiwifruit producers experiencing similar conditions to ours should focus on artificially pollinating their crops rather than relying on managed or wild insects for kiwifruit pollination. Future research should evaluate other methods of artificial pollination to determine their effectiveness, efficiency, and economics in the pollination of kiwifruit grown within the United States. [ABSTRACT FROM AUTHOR]

Plants modulate multitrophic ecological interactions, and variation in plant traits can affect these interactions. Pollinators are exposed to pathogens at flowers and acquire or transmit pathogens at different rates on different plant species, but the traits mediating those interactions are almost entirely unknown. We experimentally manipulated five plant traits that span scales including flower, inflorescence, and plant, to determine their effects on pathogen transmission between foraging bees. Specifically, we manipulated two morphological traits (corolla lip length and flower orientation within an inflorescence) and three resource distribution traits (inflorescence nectar, plant patch nectar, and plant aggregation) in tents to test how plant traits affect bee pathogen transmission. We also quantified foraging behavior and fecal deposition patterns as potential mechanisms driving differences in transmission, and assessed trait manipulation consequences for bee reproduction. We found that pathogen transmission was reduced when we trimmed the corolla lip, evenly dispersed nectar distribution within an inflorescence, or aggregated plants in space. Some traits also affected bee reproduction; tents with trimmed corollas had more larval production than control tents, and tents with evenly distributed nectar across plant patches had more larval production than tents with clumped resources. Thus, some trait manipulations both reduced transmission and increased bee microcolony reproduction, although our design does not allow us to discern whether these are related or separate effects. Taken together, our results demonstrate causal effects of several floral traits on pathogen transmission and pollinator reproduction, indicating the importance of intraspecific plant trait variation for pollinator health and population dynamics. [ABSTRACT FROM AUTHOR]

Species interaction networks, which play an important role in determining pathogen transmission and spread in ecological communities, can shift in response to agricultural landscape simplification. However, we know surprisingly little about how landscape simplification-driven changes in network structure impact epidemiological patterns. Here, we combine mathematical modelling and data from eleven bipartite plant-pollinator networks observed along a landscape simplification gradient to elucidate how changes in network structure shape disease dynamics. Our empirical data show that landscape simplification reduces pathogen prevalence in bee communities via increased diet breadth of the dominant species. Furthermore, our empirical data and theoretical model indicate that increased connectance reduces the likelihood of a disease outbreak and decreases variance in prevalence among bee species in the community, resulting in a dilution effect. Because infectious diseases are implicated in pollinator declines worldwide, a better understanding of how land use change impacts species interactions is therefore critical for conserving pollinator health.

The dilution effect hypothesis posits that increasing biodiversity reduces infectious disease transmission. Here, we propose that habitat quality might modulate this negative biodiversity–disease relationship. Habitat may influence pathogen prevalence directly by affecting host traits like nutrition and immune response (we coined the term "habitat–disease relationship" to describe this phenomenon) or indirectly by changing host biodiversity (biodiversity–disease relationship). We used a path model to test the relative strength of links between habitat, biodiversity, and pathogen prevalence in a pollinator–virus system. High‐quality habitat metrics were directly associated with viral prevalence, providing evidence for a habitat–disease relationship. However, the strength and direction of specific habitat effects on viral prevalence varied based on the characteristics of the habitat, host, and pathogen. In general, more natural area and richness of land‐cover types were directly associated with increased viral prevalence, whereas greater floral density was associated with reduced viral prevalence. More natural habitat was also indirectly associated with reduced prevalence of two key viruses (black queen cell virus and deformed wing virus) via increased pollinator species richness, providing evidence for a habitat‐mediated dilution effect on viral prevalence. Biodiversity–disease relationships varied across viruses, with the prevalence of sacbrood virus not being associated with any habitat quality or pollinator community metrics. Across all viruses and hosts, habitat–disease and biodiversity–disease paths had effects of similar magnitude on viral prevalence. Therefore, habitat quality is a key driver of variation in pathogen prevalence among communities via both direct habitat–disease and indirect biodiversity–disease pathways, though the specific patterns varied among different viruses and host species. Critically, habitat–disease relationships could either contribute to or obscure dilution effects in natural systems depending on the relative strength and direction of the habitat–disease and biodiversity–disease pathways in that host–pathogen system. Therefore, habitat may be an important driver in the complex interactions between hosts and pathogens. [ABSTRACT FROM AUTHOR]

Octopamine has broad roles within invertebrate nervous systems as a neurohormone, neurotransmitter, and neuromodulator. It orchestrates foraging behavior in many insect taxa via effects on feeding, gustatory responsiveness, and appetitive learning. Knowledge of how this biogenic amine regulates bee physiology and behavior is based largely on study of a single species, the honey bee, Apis mellifera. Until recently, its role in the foraging ecology and social organization of diverse bee taxa had been unexplored. Bumble bees (Bombus spp.) are a model for research into the neural basis of foraging and learning, but whether octopamine similarly affects sensory and cognitive performance in this genus is not known. To address this gap, we explored the effects of octopamine on gustatory responsiveness and associative learning in Bombus impatiens via conditioning of the proboscis extension reflex (PER) using a visual (color) cue. We found that octopamine had similar effects on bumble bee behavior as previously reported in honey bees; however, higher doses were required to induce these effects. At this higher dose, octopamine lowered bees’ gustatory responsiveness and appeared to enhance associative learning performance during the early phase of our experiment. Adding to recent studies on stingless bees (Meliponini), these findings support the idea that octopamine’s role in reward perception and processing is broadly conserved across Apidae, while pointing towards some differences across systems worth exploring further. [ABSTRACT FROM AUTHOR]

Judgement bias, or ‘optimism’ and ‘pessimism’, has been demonstrated across many taxa, yet the cognitive mechanisms underlying this behaviour remain unclear. In an optimism paradigm, animals are trained to an association, and, if given a positive experience, behave more favourably towards ‘ambiguous’ stimuli. We tested whether this effect could be explained by changes to stimulus response gradients by giving bees a task where their response was tested across a wider gradient of stimuli than typically tested. In line with previous work, we found that bees given a positive experience demonstrated judgement bias, being more likely to visit ambiguous stimuli. However, bees were also less likely to visit a stimulus on the other side of the rewarded stimulus (S+), and as such had a shifted stimulus response curve, showing a diminished peak shift response. In two follow-up experiments we tested the hypothesis that our manipulation altered bees’ stimulus response curves via changes to the peak shift response by reducing peak shift in controls. We found that, in support of our hypothesis, elimination of peak shift also eliminated differences between treatments. Our results point towards a cognitive explanation of ‘optimistic’ behaviour in non-human animals and offer a new paradigm for considering emotion-like states.

Pollen collection is necessary for bee survival and important for flowering plant reproduction, yet if and how pollen extraction motor routines are modified with experience is largely unknown. Here, we used an automated reward and monitoring system to evaluate modification in a common pollen-extraction routine, floral sonication. Through a series of laboratory experiments with the bumblebee, Bombus impatiens, we examined whether variation in sonication frequency and acceleration is due to instrumental learning based on rewards, a fixed behavioral response to rewards, and/or a mechanical constraint. We first investigated whether bees could learn to adjust their sonication frequency in response to pollen rewards given only for specified frequency ranges and found no evidence of instrumental learning. However, we found that absence versus receipt of a pollen reward did lead to a predictable behavioral response, which depended on bee size. Finally, we found some evidence of mechanical constraints, in that flower mass affected sonication acceleration (but not frequency) through an interaction with bee size. In general, larger bees showed more flexibility in sonication frequency and acceleration, potentially reflecting a size-based constraint on the range over which smaller bees can modify frequency and acceleration. Overall, our results show that although bees did not display instrumental learning of sonication frequency, their sonication motor routine is nevertheless flexible.

Movement of foraging animals can be used as an indirect index of habitat quality because movement patterns are expected to shift in association with resources resulting in increased time spent in high‐quality habitat. Here, we observed movement of bumblebee (Bombus impatiens) queens as an indirect metric of nesting habitat quality.At a farm in Massachusetts, we recorded flight paths by visual observation of nest‐searching B. impatiens queens in three land cover types (hay fields, meadows and forests). According to previous research at this field site, these land cover types represent both high‐quality (meadows, forests) and low‐quality habitat (hay fields) for nesting bumblebees. Using flight path data, we estimated diffusion coefficients of nest‐searching queens (i.e., the rate at which queens explore habitat during nest‐searching), as an integrative measure of movement.We found that nest‐searching queens explored meadows and forests more slowly than hay fields; we calculated diffusion coefficients of 0.198, 0.272, and 0.762 m2 s−1 for bees searching for nests in meadows, forests, and hay fields, respectively. In hay fields, nest‐searching queens had smaller step lengths and straighter flightpaths than in other land cover types.Nesting ecology of bumblebees is a key knowledge gap. We showed that nest‐searching queens search more thoroughly in areas associated with previously reported high nest densities. Therefore, our research suggests that behaviour of nest‐searching queens can also be a proxy for nesting habitat quality. Extending these methods to other Bombus species in other landscapes would be a valuable direction for future research. [ABSTRACT FROM AUTHOR]

Crithidia bombi is a trypanosomatid parasite that infects several species of bumble bees (Bombus spp.), by adhering to their intestinal tract. Crithidia bombi infection impairs learning and reduces survival of workers and the fitness of overwintering queens. Although there is extensive research on the ecology of this host-pathogen system, we understand far less about the mechanisms that mediate internal infection dynamics. Crithidia bombi infects hosts by attaching to the hindgut via the flagellum, and one previous study found that a nectar secondary compound removed the flagellum, preventing attachment. However, approaches that allow more detailed observation of parasite attachment and growth would allow us to better understand factors mediating this host-pathogen relationship. We established techniques for genetic manipulation and visualization of cultured C. bombi. Using constructs established for Crithidia fasciculata, we successfully generated C. bombi cells expressing ectopic fluorescent transgenes using two different selectable markers. To our knowledge, this is the first genetic modification of this species. We also introduced constructs that label the mitochondrion and nucleus of the parasite, showing that subcellular targeting signals can function across parasite species to highlight specific organelles. Finally, we visualized fluorescently tagged parasites in vitro in both their swimming and attached forms, and in vivo in bumble bee (Bombus impatiens) hosts. Expanding our cell and molecular toolkit for C. bombi will help us better understand how factors such as host diet, immune system, and physiology mediate outcomes of infection by these common parasites. [Display omitted] • First described method for genetic modification of the bee parasite Crithidia bombi. • Ectopic transgenes can be introduced using multiple selectable markers. • Cytoplasmic and organelle markers can be visualized in swimming and attached developmental forms. • Simplified method for culture adaptation of parasites from bees for subsequent modification. • Imaging of fluorescent C. bombi parasites in vivo in the bumble bee Bombus impatiens. [ABSTRACT FROM AUTHOR]

The spread of parasites is one of the primary drivers of population decline of both managed and wild bees. Several bee parasites are transmitted by the shared use of flowers, turning floral resources into potential disease hotspots. However, we know little about how floral morphology and floral species identity affect different steps of the transmission process. Here, we used the gut parasite Crithidia bombi and its primary host, bumble bees (Bombus spp.), to examine whether floral traits or species identity better predict three basic steps of parasite transmission on flowers: feces deposition on flowers, survival of the parasite on flowers, and acquisition by a new host. We also identified which traits and/or species were most strongly associated with each step in the transmission process. We found that both trait‐ and species‐based models fit the data on deposition of feces and survival of C. bombi on flowers, but that species‐based models provided a better fit compared with trait‐based ones. However, trait‐based models were better at predicting the acquisition of C. bombi on flowers. Although different species tended to support higher fecal deposition or parasite survival, we found that floral shape provided explanatory power for each of the transmission steps. When we assessed overall transmission potential, floral shape had the largest explanatory effect, with wider, shorter flowers promoting higher transmission. Taken together, our results highlight the importance of flower species identity and floral traits in disease transmission dynamics of bee parasites, and floral shape as an important predictor of overall transmission potential. Identifying traits associated with transmission potential may help us create seed mix that presents lower parasite transmission risk for bees for use in pollinator habitat. [ABSTRACT FROM AUTHOR]

While most models of decision-making assume that individuals assign options absolute values, animals often assess options comparatively, violating principles of economic rationality. Such 'irrational' preferences are especially common when two rewards vary along multiple dimensions of quality and a third, 'decoy' option is available. Bumblebees are models of decision-making, yet whether they are subject to decoy effects is unknown. We addressed this question using bumblebees ( Bombus impatiens ) choosing between flowers that varied in their nectar concentration and reward rate. We first gave bees a choice between two flower types, one higher in concentration and the other higher in reward rate. Bees were then given a choice between these flowers and either a 'concentration' or 'rate' decoy, designed to be asymmetrically dominated on each axis. The rate decoy increased bees' preference in the expected direction, while the concentration decoy did not. In a second experiment, we manipulated choices along two single reward dimensions to test whether this discrepancy was explained by differences in how concentration versus reward rate were evaluated. We found that low-concentration decoys increased bees' preference for the medium option as predicted, whereas low-rate decoys had no effect. Our results suggest that both low- and high-value flowers can influence pollinator preferences in ways previously unconsidered.

Certain diets can benefit bee health by reducing pathogens, but the mechanism(s) driving these medicinal effects are largely unexplored. Recent research found that sunflower (Helianthus annuus) pollen reduces the gut pathogen Crithidia bombi in the common eastern bumblebee (Bombus impatiens). Here, we tested the effects of sunflower pollen and infection on two bee immune metrics to determine whether sunflower pollen diet drives changes in host immunity that can explain this medicinal effect. Bees were infected with C. bombi or not and given either sunflower or wildflower pollen. Subsequently, bees received a benign immune challenge or were left naive to test the induced and constitutive immune responses, respectively. We measured haemolymph phenoloxidase activity, involved in the melanization cascade, and antibacterial activity. Sunflower pollen reduced C. bombi infection, but we found no significant pollen diet effect on either immune measure. Phenoloxidase activity was also not affected by C. bombi infection status; however, uninfected bees were more likely to have measurable constitutive antibacterial activity, while infected bees had higher induced antibacterial activity. Overall, we found that sunflower pollen does not significantly affect the immune responses we measured, suggesting that the mechanisms underlying its medicinal effect do not involve these bee immune parameters. This article is part of the theme issue 'Natural processes influencing pollinator health: from chemistry to landscapes'. [ABSTRACT FROM AUTHOR]

Neonicotinoid pesticides in the nectar and pollen of managed crops and wildflowers contribute to the global declines of bees. These chemicals can have detrimental effects on bees' physiology, behaviour and reproduction. Floral nectar also contains secondary chemistry with its own effects on bee health. How nectar secondary chemistry may act additively or synergistically with neonicotinoids is unknown.Here, we asked how an acute exposure to a common neonicotinoid, imidacloprid (IMD) affected the longevity, immune function and behaviour of bumble bee Bombus impatiens workers maintained on diets enriched with one of three nectar secondary metabolites (NSMs; the alkaloid caffeine, the terpenoid thymol or the cardiac glycoside digoxin). A factorial design allowed us to assess the potential for additive and interactive effects of each NSM and IMD combination on multiple health outcomes.Without IMD exposure, different dietary NSMs each had positive effects on life span (caffeine), immune function (digoxin) and activity levels (caffeine, thymol), although these came with trade‐offs. A single sublethal IMD exposure overshadowed these NSM effects, and in two cases, an NSM‐enriched diet magnified the negative effects of pesticide exposure.In summary, we show that even a single acute exposure to a pesticide has the potential to reshape interactions between pollinators and plants mediated by nectar secondary chemistry. Read the free Plain Language Summary for this article on the Journal blog. [ABSTRACT FROM AUTHOR]

Flying insects encounter a considerable amount of multisensorial information during their first foraging trip and need to extract and process relevant cues to efficiently navigate their environment and locate food sources. Previous studies used static stimuli to investigate visual information processing during flight and the role of floral features on detection, landing, and flower handling behaviours. However, bees come across visual information sequentially, while sampling the visual scene and presenting visual features that change after landing would allow further understanding of the chronological aspect of visual information processing. Here, we describe a new methodology that uses the ShadowSense™ Multitouch technology to present interactive floral displays where a change in visual features is triggered upon a bee's landing. Two colonies of flower naïve bumblebees (Bombus impatiens) were exposed to unicoloured and bicoloured unrewarded floral images in which floral guides in the form of dummy stamens were added, removed, or remained unchanged in the centre upon landing. Our findings confirm preference by flower‐naïve bumblebees for bicoloured flowers and corroborate that small central visual guides direct the place of landing. Therefore, we establish proof‐of‐concept of this new methodology for bee research by reproducing previously demonstrated behaviours and by reporting that bees react to a change in visual information on the touchscreen. To our knowledge, this is the first research providing a touchscreen technology that can reliably be used with bees. While its efficiency with similar‐sized insects is yet to be confirmed, this technology provides new approaches for research on visual information processing as well as various behaviours in insects. [ABSTRACT FROM AUTHOR]

• Warmer temperatures or drier conditions accelerated bumble bee water loss. • Warmer temperatures, but not drier conditions, resulted in earlier mortality. • Bumble bees had less water in their body at mortality under low humidity. • Larger bees could withstand more stressful conditions. • Colonies varied in their water balance traits. Ongoing climate change has increased temperatures and the frequency of droughts in many parts of the world, potentially intensifying the desiccation risk for insects. Because resisting desiccation becomes more difficult at higher temperatures and lower humidity, avoiding water loss is a key challenge facing terrestrial insects. However, few studies have examined the interactive effects of temperature and environmental humidity on desiccation resistance in insects. Such studies on bees (Hymenoptera: Apoidea: Anthophila) are especially rare, despite their ecological and economic importance. Here, we crossed temperature (20, 25, and 30 °C) with humidity (<5, 50, >95 % RH) manipulations and measured time to mortality, water loss rates, and the water content at mortality of bumble bees (Bombus impatiens). We found that both higher temperature and lower humidity increased water loss rates, while warmer temperatures reduced survival time and lower humidity decreased water content at mortality. Additionally, we observed large intraspecific variation in water balance traits between colonies, and larger individuals survived longer and could tolerate more water loss before mortality. This study raises important questions about the mechanisms underpinning water loss in bumble bees and suggests that frequent access to nectar may be especially important for bumble bees' water balance and survival in a warming and drying climate. [ABSTRACT FROM AUTHOR]

Although bee visitation rate to flowers is often used to assess both pollination services and bee abundance, the abundance of social species needs to be assessed by quantifying the number colonies instead of the number of foraging individuals. Because accurately quantifying the number of wild bee colonies can be difficult, the relationship of visitation rates provided by foragers and the abundance of colonies contributing those foragers from the surrounding landscape is poorly documented for social species. Here, we use genetic methods and statistical inference to estimate the abundance of wild colonies of Bombus impatiens in the surrounding landscape providing foragers to 30 commercial pumpkin (Cucurbita pepo) fields over 4 yr across a 13,000‐km2 area in Pennsylvania, USA. We show that the abundance of wild colonies in the surrounding landscape providing foragers per field (colony abundance per field) ranges from 291 to 829 colonies per field. Furthermore, colony abundance per field has been stable across years, counties, and field size, resulting in a dilution of foragers from the available colonies across larger fields. Wild colony abundance, when scaled by field size, by expressing colony abundance on a per hectare basis, is predictive of visitation rate. Thus, we document the relationship of wild colony abundance per hectare to visitation rate at a level that provides sufficient pollination to a highly pollinator‐dependent crop. As expected, genetic differentiation among sampled populations is essentially non‐existent across different fields or regions, suggesting a panmictic population. Although many Bombus species are in decline, we document abundant and genetically resilient wild populations of B. impatiens associated with a mass‐flowering crop under current agricultural practices and provide baseline information needed to monitor these wild populations at a time when they face similar stressors implicated in the decline of congenerics. [ABSTRACT FROM AUTHOR]

Animals develop food preferences based on taste, nutritional quality and to avoid environmental toxins. Yet, measuring preferences in an experimental setting can be challenging since ecologically realistic assays can be time consuming, while simplified assays may not capture natural sampling behavior. Field realism is a particular challenge when studying behavioral responses to environmental toxins in lab-based assays, given that toxins can themselves impact sampling behavior, masking our ability to detect preferences. We address these challenges by comparing different experimental methods for measuring sucrose concentration preference in bumble bees (Bombus impatiens), evaluating the utility of two preference chamber-based methods (ad libitum versus a novel restricted-sampling assay) in replicating bees' preferences when they fly freely between artificial flowers in a foraging arena. We find that the restricted-sampling method matched a free-flying scenario more closely than the ad libitum protocol, and we advocate for expanded use of this approach, given its ease of implementation. We then performed a second experiment using the new protocol to ask whether consuming the neonicotinoid pesticide imidacloprid, known to suppress feeding motivation, interfered with the expression of sucrose preferences. After consuming imidacloprid, bees were less likely to choose the higher-quality sucrose even as they gained experience with both options. Thus, we provide evidence that pesticides interfere with bees' ability to discriminate between floral rewards that differ in value. This work highlights a simple protocol for assessing realistic foraging preferences in bees and provides an efficient way for researchers to measure the impacts of anthropogenic factors on preference expression. [ABSTRACT FROM AUTHOR]

Trait variation can have important consequences for the outcomes of species interactions. Even though some traits vary as much within species as across related species, models and empirical studies typically do not consider the role of intraspecific trait variation for processes such as disease transmission. For example, many pollinator species are in decline because of a variety of stressors including pathogens, but the role of intraspecific trait variation in mediating disease dynamics is rarely considered. For example, pollinator body size could affect pathogen transmission via differences in resistance, foraging behavior and physiology. We tested effects of body size on pollinator pathogen transmission using the common eastern bumble bee Bombus impatiens in field tents, introducing an infected "donor" microcolony of large or small workers with an uninfected average‐sized "recipient" microcolony and allowing bees to forage for 9–16 d. Small donor bees had nearly 50% higher infection intensity (cells/0.02 μL) than large donor bees, but large donor bees were twice as likely to transmit Crithidia bombi to recipient bees. Both behavioral and physiological mechanisms may underlie this apparent paradox. Compared to small bees, large bees foraged more and produced more feces; simulations showed that foraging and defecation rates together had stronger effects on transmission than did donor infection intensity. Thus, effects of bee size on contact rates and pathogen supply may play significant roles in disease transmission, demonstrating the multifaceted impacts of traits on transmission dynamics. [ABSTRACT FROM AUTHOR]

Understanding habitat quality is central to understanding the distributions of species on the landscape, as well as to conserving and restoring at‐risk species. Although it is well known that many species require different resources throughout their life cycles, pollinator conservation efforts focus almost exclusively on forage resources. In this study, we evaluate nesting habitat for bumble bees by locating nests directly on the landscape. We compared colony density and colony reproductive output for Bombus impatiens, the common eastern bumble bee, across three different land cover types (hay fields, meadows, and forests). We also assessed nesting habitat associations for all Bombus nests located during surveys to tease apart species‐specific patterns of habitat use. We found that B. impatiens nested under the ground in two natural land cover types, forests, and meadows, but found no B. impatiens nests in hay fields. Though B. impatiens nested at similar densities in both meadows and forests, colonies in forests had much higher reproductive output. In contrast, B. griseocollis tended to nest on the surface of the ground and was almost always found in meadows. B. perplexis was the only species to nest in all three habitat types, including hay fields. For some bumble bee species in this system, meadows, the habitat type with abundant forage resources, may be sufficient to maintain them throughout their life cycles. However, B. impatiens might benefit from heterogeneous landscapes with forests and meadows. Results for B. impatiens emphasize the longstanding notion that habitat use is not always positively correlated with habitat quality (as measured by reproductive output). Our results also show that habitat selection by bumble bees at one spatial scale may be influenced by resources at other scales. Finally, we demonstrate the feasibility of direct nest searches for understanding bumble bee distribution and ecology. [ABSTRACT FROM AUTHOR]

Abstract Although bee visitation rate to flowers is often used to assess both pollination services and bee abundance, the abundance of social species needs to be assessed by quantifying the number colonies instead of the number of foraging individuals. Because accurately quantifying the number of wild bee colonies can be difficult, the relationship of visitation rates provided by foragers and the abundance of colonies contributing those foragers from the surrounding landscape is poorly documented for social species. Here, we use genetic methods and statistical inference to estimate the abundance of wild colonies of Bombus impatiens in the surrounding landscape providing foragers to 30 commercial pumpkin (Cucurbita pepo) fields over 4 yr across a 13,000‐km2 area in Pennsylvania, USA. We show that the abundance of wild colonies in the surrounding landscape providing foragers per field (colony abundance per field) ranges from 291 to 829 colonies per field. Furthermore, colony abundance per field has been stable across years, counties, and field size, resulting in a dilution of foragers from the available colonies across larger fields. Wild colony abundance, when scaled by field size, by expressing colony abundance on a per hectare basis, is predictive of visitation rate. Thus, we document the relationship of wild colony abundance per hectare to visitation rate at a level that provides sufficient pollination to a highly pollinator‐dependent crop. As expected, genetic differentiation among sampled populations is essentially non‐existent across different fields or regions, suggesting a panmictic population. Although many Bombus species are in decline, we document abundant and genetically resilient wild populations of B. impatiens associated with a mass‐flowering crop under current agricultural practices and provide baseline information needed to monitor these wild populations at a time when they face similar stressors implicated in the decline of congenerics.

Abstract Understanding habitat quality is central to understanding the distributions of species on the landscape, as well as to conserving and restoring at‐risk species. Although it is well known that many species require different resources throughout their life cycles, pollinator conservation efforts focus almost exclusively on forage resources. In this study, we evaluate nesting habitat for bumble bees by locating nests directly on the landscape. We compared colony density and colony reproductive output for Bombus impatiens, the common eastern bumble bee, across three different land cover types (hay fields, meadows, and forests). We also assessed nesting habitat associations for all Bombus nests located during surveys to tease apart species‐specific patterns of habitat use. We found that B. impatiens nested under the ground in two natural land cover types, forests, and meadows, but found no B. impatiens nests in hay fields. Though B. impatiens nested at similar densities in both meadows and forests, colonies in forests had much higher reproductive output. In contrast, B. griseocollis tended to nest on the surface of the ground and was almost always found in meadows. B. perplexis was the only species to nest in all three habitat types, including hay fields. For some bumble bee species in this system, meadows, the habitat type with abundant forage resources, may be sufficient to maintain them throughout their life cycles. However, B. impatiens might benefit from heterogeneous landscapes with forests and meadows. Results for B. impatiens emphasize the longstanding notion that habitat use is not always positively correlated with habitat quality (as measured by reproductive output). Our results also show that habitat selection by bumble bees at one spatial scale may be influenced by resources at other scales. Finally, we demonstrate the feasibility of direct nest searches for understanding bumble bee distribution and ecology.

Insecticide use and insufficient forage are two of the leading stressors to pollinators in agroecosystems. While these factors have been well studied individually, the experimental designs do not reflect real-world conditions where insecticide exposure and lack of forage occur simultaneously and could interactively suppress pollinator health. Using outdoor enclosures, we tested the effects of insecticides (imidacloprid + lambda-cyhalothrin) and non-crop forage (clover) in a factorial design, measuring the survival, behaviour and performance of bumblebees (Bombus impatiens), as well as pollination of the focal crop, watermelon. Colony survival was synergistically reduced to 17% in watermelon alone + insecticides (survival was 100% in all other treatments). However, behavioural shifts in foraging were mainly owing to insecticides (e.g. 95% reduced visitation rate to watermelon flowers), while impacts on hive performance were primarily driven by clover presence (e.g. 374% increase in the number of live eggs). Insecticide-mediated reductions in foraging decreased crop pollination (fruit set) by 32%. Altogether, these data indicate that both insecticides and non-crop forage play integral roles in shaping pollinator health in agricultural landscapes, but the relative importance and interaction of these two factors depend on which aspect of 'health' is being considered. [ABSTRACT FROM AUTHOR]

Insect pollinators readily learn olfactory cues, and this is expected to select for 'honest signals' that provide reliable information about floral rewards. However, plants might alternatively produce signals that exploit pollinators' sensory biases, thereby relaxing selection for signal honesty. We examined the innate and learned preferences of Bombus impatiens for Mimulus guttatus floral scent phenotypes corresponding to different levels of pollen rewards in the presence and absence of the innately attractive floral volatile compound β-trans-bergamotene. Bees learned to prefer honest signals after foraging on live M. guttatus flowers, but only exhibited this preference when presented floral scent phenotypes that did not include β-trans-bergamotene. Our results suggest that a sensory bias for β-trans-bergamotene overrides the ability of B. impatiens to use honest signals when foraging on M. guttatus. This may represent a deceptive pollination strategy that allows plants to minimize investment in costly rewards without incurring reduced rates of pollinator visitation. [ABSTRACT FROM AUTHOR]

1. Social insect castes and sexes differ in many ways, including morphology, behavior, and sometimes ploidy level. Recent studies have found that consuming sunflower pollen reduces the gut pathogen Crithidia bombi in workers of the common eastern bumble bee (Bombus impatiens). Here, this work is extended to the reproductive individuals that represent colony fitness – males and queens – to assess if the medicinal effects of sunflower pollen vary with bee caste and sex. 2. This study examined the effect of sunflower pollen compared to a diverse wildflower pollen mix on infection in worker, male, and daughter queen commercial B. impatiens. Bees were infected, fed either sunflower pollen or wildflower pollen for 7 days, and then infection levels were assessed. 3. Compared to wildflower pollen, sunflower pollen dramatically reduced Crithidia infection in workers and daughter queens, but not males. Infection levels were very low for both diets in males; this could be due to low pollen consumption or other mechanisms. 4. Reducing Crithidia infection in young queens before they undergo hibernation is important for population dynamics since infected queens are less likely to survive hibernation, and those that do are less likely to successfully establish a nest the following spring. Because sunflowers bloom in late summer when new queens are emerging, sunflowers could provide an important dietary component for queens during this critical life stage. Deepening our understanding of how diet impacts pathogens in reproductive bees, as well as workers, is crucial to maintain healthy pollinator populations. [ABSTRACT FROM AUTHOR]

Parasites are commonly cited as one of the causes of population declines for both managed and wild bees. Epidemiological models sometimes assume that increasing the proportion of infected individuals in a group should increase transmission. However, social insects exhibit behaviors and traits which can dampen the link between parasite pressure and disease spread. Understanding patterns of parasite transmission within colonies of social bees has important implications for how to control diseases within those colonies, and potentially the broader pollinator community. We used bumble bees (Bombus impatiens Cresson) (Hymenoptera: Apidae) and western honey bees (Apis mellifera L.) (Hymenoptera: Apidae) infected with the gut parasites Crithidia bombi (Lipa & Triggiani) (Trypanosomatida: Trypanosomatidae) and Nosema ceranae (Fries et al.) (Dissociodihaplophasida: Nosematidae), respectively, to understand how the initial proportion of infected individuals impacts within-colony spread and intensity of infection of the parasites. In bumble bees, we found that higher initial parasite prevalence increased both the final prevalence and intensity of infection of C. bombi. In honey bees, higher initial prevalence increased the intensity of infection in individual bees, but not the final prevalence of N. ceranae. Measures that reduce the probability of workers bringing parasites back to the nest may have implications for how to control transmission and/or severity of infection and disease outbreaks, which could also have important consequences for controlling disease spread back into the broader bee community. [ABSTRACT FROM AUTHOR]

Pityopsis ruthii (Small) Small (Asteraceae: Asterales) is a federally endangered aster that only grows along short stretches of the Hiwassee and Ocoee Rivers in southeastern Tennessee, U.S.A. The species is self-incompatible and genetic drift and inbreeding depression have been reported. Multiple investigators have observed highly variable sexual reproductive capacity of P. ruthii. To better understand which insect species may be contributing roles as potential pollinators, floral visitation to P. ruthii was assessed at in situ and ex situ locations. Forty-seven insect species in 15 families and 3 orders were recovered across 2 years during more than 70 hours of observation and specimen collection. Megachile brevis Say (Megachilidae: Hymenoptera) carried the most pollen, yet just three specimens were collected, all from the ex situ location. In both years, Halictidae were common at the ex situ location and were infrequently collected at the in situ location. Apis mellifera L. (Apidae: Hymenoptera) and Bombus impatiens Cresson (Apidae: Hymenoptera) were commonly observed at in situ locations in both years, and in 2013 at the ex situ location. Toxomerus geminatus (Say) (Syrphidae: Diptera) was the most abundant floral visitor in both years but carried very little pollen. Floral visiting lepidopteran species carried no pollen. Greater numbers of P. ruthii inflorescences did not reliably influence Apoidea or Syrphidae floral visitor abundance in either the in situ or ex situ locations. Assessments of filled seeds revealed low germination rates. Hybridization between geographically disparate parent plants propagated from the distal ranges of Hiwassee River distribution yielded an average of 33 percent germination, validating evidence of inbreeding depression. Knowledge of P. ruthii floral visitors at in situ and ex situ locations can be employed to assist breeding efforts intended to increase genetic diversity through ongoing efforts in plant re-introduction and habitat management. [ABSTRACT FROM AUTHOR]

Herbivory can induce chemical changes throughout plant tissues including flowers, which could affect pollinator-pathogen interactions. Pollen is highly defended compared to nectar, but no study has examined whether herbivory affects pollen chemistry. We assessed the effects of leaf herbivory on nectar and pollen alkaloids in Nicotiana tabacum, and how herbivory-induced changes in nectar and pollen affect pollinator-pathogen interactions. We damaged leaves of Nicotiana tabacum using the specialist herbivore Manduca sexta and compared nicotine and anabasine concentrations in nectar and pollen. We then pooled nectar and pollen by collection periods (within and after one month of flowering), fed them in separate experiments to bumble bees (Bombus impatiens) infected with the gut pathogen Crithidia bombi, and assessed infections after seven days. We did not detect alkaloids in nectar, and leaf damage did not alter the effect of nectar on Crithidia counts. In pollen, herbivory induced higher concentrations of anabasine but not nicotine, and alkaloid concentrations rose and then fell as a function of days since flowering. Bees fed pollen from damaged plants had Crithidia counts 15 times higher than bees fed pollen from undamaged plants, but only when pollen was collected after one month of flowering, indicating that both damage and time since flowering affected interaction outcomes. Within undamaged treatments, bees fed late-collected pollen had Crithidia counts 10 times lower than bees fed early-collected pollen, also indicating the importance of time since flowering. Our results emphasize the role of herbivores in shaping pollen chemistry, with consequences for interactions between pollinators and their pathogens. [ABSTRACT FROM AUTHOR]

Recent studies have documented that social bees can use heterospecific information to find or avoid food resources, but little is known about whether bees gain information from heterospecifics about predation risk. We report the first detailed field tests in bees of hetero- and conspecific avoidance of olfactory information associated with predation. We determined whether Apis mellifera and Bombus impatiens would respond either to hetero- or conspecific haemolymph as an indication of a predation event, or to sting gland contents, which provide an alarm pheromone in honeybees and in many other social Hymenoptera. Bombus impatiens avoided their own haemolymph and A. mellifera haemolymph in foraging arena choice experiments. Bombus impatiens did not respond to A. mellifera alarm pheromone or to the odour of conspecific sting gland. In field experiments, A. mellifera avoided their own haemolymph and their own sting alarm pheromone, but did not avoid the haemolymph or sting gland contents of B. impatiens or native bumblebees (Bombus vosnesenskii) that regularly foraged around their hives. One factor behind the response of B. impatiens to heterospecific cues of predation may be its habit of solitary foraging, which may lead to more interactions with heterospecifics than would social foraging in which bees recruit nestmates to resources. © 2012 The Association for the Study of Animal Behaviour.

Many pollinator species are declining due to a variety of interacting stressors including pathogens, sparking interest in understanding factors that could mitigate these outcomes. Diet can affect host-pathogen interactions by changing nutritional reserves or providing bioactive secondary chemicals. Recent work found that sunflower pollen (Helianthus annuus) dramatically reduced cell counts of the gut pathogen Crithidia bombi in bumble bee workers (Bombus impatiens), but the mechanism underlying this effect is unknown. Here we analyzed methanolic extracts of sunflower pollen by LC-MS and identified triscoumaroyl spermidines as the major secondary metabolite components, along with a flavonoid quercetin-3-O-hexoside and a quercetin-3-O-(6-O-malonyl)-hexoside. We then tested the effect of triscoumaroyl spermidine and rutin (as a proxy for quercetin glycosides) on Crithidia infection in B. impatiens, compared to buckwheat pollen (Fagopyrum esculentum) as a negative control and sunflower pollen as a positive control. In addition, we tested the effect of nine fatty acids from sunflower pollen individually and in combination using similar methods. Although sunflower pollen consistently reduced Crithidia relative to control pollen, none of the compounds we tested had significant effects. In addition, diet treatments did not affect mortality, or sucrose or pollen consumption. Thus, the mechanisms underlying the medicinal effect of sunflower are still unknown; future work could use bioactivity-guided fractionation to more efficiently target compounds of interest, and explore non-chemical mechanisms. Ultimately, identifying the mechanism underlying the effect of sunflower pollen on pathogens will open up new avenues for managing bee health. [ABSTRACT FROM AUTHOR]

Floral sonication is a behaviour in which certain bees generate substrate-borne vibrations by contracting their flight muscles in order to extract pollen from poricidal anthers. Sonication vibrations, or 'buzzes', also contain a corresponding audible sound that results from vibrations radiating from the exoskeleton into the surrounding air. Acoustic recordings are often used as a proxy for analysis of floral sonication vibrations because they are more accessible to recording with conventional equipment such as microphones. However, the extent to which salient parameters of buzzes are reflected in the airborne components has not been experimentally verified. We examined correspondence in three properties (duration, frequency and amplitude) simultaneously recorded with acoustic and vibrational methods from freely foraging bumblebees. Duration and frequency are faithfully quantified from airborne recordings; however, two measures of acoustic amplitude (relative peak amplitude and sound pressure level) are not correlated with vibrational amplitude. Our findings validate acoustic recordings as a method to describe temporal and spectral components of floral sonication vibrations, but we caution against using acoustic measures of amplitude as proxies for the true vibrational power. [ABSTRACT FROM AUTHOR]

1. Several bee species are declining due to multiple factors, including pathogens. Ingestion of sunflower (Helianthus annuus) pollen can dramatically reduce the bumble bee gut pathogen Crithidia bombi, but little is known about how timing and exposure to sunflower pollen consumption affects pathogen load. 2. Two experiments were carried out to investigate how exposure to sunflower pollen relative to pathogen exposure affects Crithidia bombi in Bombus impatiens. Foraging trials with pollen‐producing and male‐sterile (pollen absent) sunflower lines were performed to investigate whether sunflower pollen affected pathogen transmission in a single foraging bout, and 7‐day laboratory trials were done to investigate whether timing and duration of exposure to sunflower pollen after infection affected C. bombi. 3. In foraging trials, pollen presence on inflorescences inoculated with C. bombi did not affect transmission (pathogen cell counts of foraging workers) 1 week later, suggesting that a brief experience with sunflower pollen concurrent with pathogen exposure is insufficient to reduce infection. In laboratory trials, consuming sunflower pollen for the first 3.5 days or all 7 days after infection reduced cell counts compared with a negative control pollen, but consuming sunflower pollen starting 3.5 days after infection did not. Consuming sunflower pollen for 7 days was significantly and substantially more effective than any other treatment. Thus, both duration and timing of exposure to sunflower pollen may affect pathogen load. 4. These results are important for understanding ecological disease dynamics in natural settings with free‐flying bumble bees, and may inform decisions about using medicinal diets to manage bumble bee health commercially. [ABSTRACT FROM AUTHOR]