Male Bumblebees Research Articles

Sunflower pollen reduces a gut pathogen in worker and queen but not male bumble bees

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.

Hyperthermic stress resistance of bumblebee males: test case of Belgian species

Thermotolerance has often been linked to species distribution for a diverse range of organisms. In the context of climate change, assessing heat resistance ability is useful for understanding potential future range shifts and the physiological response of populations. As bumblebee (Bombus) populations have been declining for several decades with several documented range shifts, an assessment of the hyperthermic resistance of species is urgently needed. In this study, we measure in males the heat resistance of ten bumblebee species living in temperate regions (northwestern Europe) with a static temperature methodology to evaluate the time before heat stupor (THS) which corresponds to a chill coma. Our results on heat stress resistance show that not all species are affected in the same way to heat stress. The most widespread species, B. terrestris (median THS 395 min) and B. lucorum (median THS 257 min) are the least sensitive to hyperthermic stress. The resistance time of bumblebee males is up to 10 times longer than the THS for declining species such as B. jonellus (median THS 48 min) and B. magnus (median THS 58 min). We highlight the high interspecific variability of heat resistance in a morphologically homogeneous genus such as bumblebees. From a conservation point of view, our research highlights the urgency for assessing the heat resistance of different species since each one can display a species-specific thermal sensitivity that is likely linked to a risk of decline in the case of heat waves.

Nectar preferences in male bumblebees

Social insect males are relatively understudied, but knowledge of their biology is increasingly important for conservation of declining groups. Bumblebees are important pollinators in temperate, sub-arctic, and Alpine regions, but many species are in decline across the globe. Agri-environment schemes have been designed to support female bumblebees, but free-living males may also need support. Male bumblebees have an energetically expensive lifestyle, and so understanding their foraging choices may provide a first step towards designing interventions to support them in the wild. Using a series of controlled laboratory choice experiments, we demonstrate that males prefer sugar concentrations of between 50 and 60% when under either simple (binary) or complex (octanary) choice conditions when presented with a broad range of sugar concentrations. When presented with concentrations within this preferred range, males exhibit a clear preference for sugar solutions of 56% w/w sucrose when solutions differ by 4%. These results suggest that males maximise their rate of energy intake, as predicted by theoretical models, rather than minimise their water consumption. In the future, male preferences for high sugar concentrations can be combined with knowledge of baseline nectar concentrations and flower phenology to maximise the value of conservation interventions for this neglected sex.

Thoracobombus from Mexico: a description of the male species-specific cephalic labial gland secretions

The male cephalic labial gland secretions of bumblebees are known to be species-specific semiochemicals. These secretions that are involved in bumblebee pre-mating recognition provide efficient diagnostic characters for species delimitation. The subgenus Thoracobombus is the largest group of bumblebees and is found in the Palearctic, Nearctic, and Neotropical regions. Here, the cephalic labial gland secretion (CLGS) composition of six Mexican Thoracobombus bumblebee species are analyzed: Bombus diligens, B. medius, B. mexicanus, B. pensylvanicus, B. trinominatus, and B. weisi. Our results suggest the presence of two new potential species into the formerly recognized B. weisi as well as one new potential species in the taxon presently identified as B. pensylvanicus. Moreover, the male of B. pensylvanicus, known to congregate at nest sites awaiting the emergence of virgin queens, is characterized by low concentrations of the C16 component. This observation raises the possibility that courtship behavior as well as environmental constraints could affect the role of the male bumblebees’ CLGS.

One size does not fit all: Caste and sex differences in the response of bumblebees (Bombus impatiens) to chronic oral neonicotinoid exposure.

Neonicotinoid insecticides have been implicated in the rapid global decline of bumblebees over recent years, particularly in agricultural and urban areas. While there is much known about neonicotinoid toxicity effects at the colony stage of the bumblebee annual cycle, far less is known about such effects at other stages critical for the maintenance of wild populations. In the present work, individual-based feeding assays were used to show that chronic consumption of the widely used neonicotinoid clothianidin at a field-realistic average rate of 3.6 and 4.0 ng/g·bee/day reduces survival of queen and male bumblebees, respectively, within a 7-day period. In contrast, worker survival was unaffected at a similar consumption rate of 3.9 ng/g·bee/day. To test the hypothesis that males have a lower tolerance for oral clothianidin exposure than workers due to their haploid genetic status, RNAseq analysis was used to compare the transcriptomic responses of workers and males to chronic intake of clothianidin at a sub-lethal dose of 0.37ng/bee/day for 5 days. Surprisingly, clothianidin consumption only altered the expression of 19 putative detoxification genes in a sex-specific manner, with 11/19 genes showing increased expression in workers. Sub-lethal clothianidin exposure also altered the expression of 40 genes associated with other major biological functions, including locomotion, reproduction, and immunity. Collectively, these results suggest that chronic oral toxicity effects of neonicotinoids are greatest during mating and nest establishment phases of the bumblebee life cycle. Chronic oral toxicity testing on males and queens is therefore required in order to fully assess the impact of neonicotinoids on wild bumblebee populations.

Variability in Sexual Pheromones Questions their Role in Bumblebee Pre-Mating Recognition System.

Sex-specific chemical secretions have been widely used as diagnostic characters in chemotaxonomy. The taxonomically confused group of bumblebees has reaped the benefit of this approach through the analyses of cephalic labial gland secretions (CLGS). Most of currently available CLGS descriptions concern species from the West-Palearctic region but few from the New World. Here, the CLGS of four East-Palearctic species Bombus deuteronymus, B. filchnerae, B. humilis, and B. exil (subgenus Thoracobombus) are analysed. Our results show high levels of variability in the major compounds in B. exil. In contrast, we describe a low differentiation in CLGS compounds between B. filchnerae and its phylogenetically closely related taxon B. muscorum. Moreover, the chemical profiles of B. filchnerae and B. muscorum are characterized by low concentrations of the C16 component, which is found in higher concentrations in the other Thoracobombus species. This raises the possibility that courtship behavior as well as environmental constraints could affect the role of the bumblebee males' CLGS.

Exploring the predation of UK bumblebees (Apidae, Bombus spp.) by the invasive pitcher plant Sarracenia purpurea: examining the effects of annual variation, seasonal variation, plant density and bumblebee gender

Invasive carnivorous plant species can impact the native invertebrate communities on which they prey. This article explores the predation of native UK bumblebees (Bombus spp.) by the invasive pitcher plant species Sarracenia purpurea and discusses the potential effect of S. purpurea on native bumblebees. Specifically, it evaluates whether the extent to which bumblebees are captured varies (i) over successive years, (ii) across June and July, (iii) with density of distribution of pitchers or (iv) with bumblebee gender. Pitcher contents were examined from an established population of Sarracenia purpurea growing in Dorset, UK. Results show that the total extent to which bumblebees were captured differed over the years 2012–2014 inclusive. A 1-year study in 2013 showed that more bumblebees were caught in July than in June and more bumblebees were captured when pitchers grew at high density. Results from 2013 also showed that more pitchers caught more than one bumblebee than would be expected based on a normal probability distribution and that this phenomenon affects female and male bumblebees equally. We discuss possible reasons for these results including that the bumblebees may be using S. purpurea as a resource. Further work is required to establish the exact underpinning mechanisms and the relative roles of plant and bumblebee behaviour within the relationship. Such interaction complexity may have consequences for consideration in invasive carnivorous plant management.

Male bumblebees perform learning flights on leaving a flower but not when leaving their nest.

Female bees and wasps demonstrate, through their performance of elaborate learning flights, when and where they memorise features of a significant site. An important feature of these flights is that the insects look back to fixate the site that they are leaving. Females, which forage for nectar and pollen and return with it to the nest, execute learning flights on their initial departure from both their nest and newly discovered flowers. To our knowledge, these flights have so far only been studied in females. Here, we describe and analyse putative learning flights observed in male bumblebees Bombus terrestris L. Once male bumblebees are mature, they leave their nest for good and fend for themselves. We show that, unlike female foragers, males always fly directly away from their nest, without looking back, in keeping with their indifference to their natal nest. In contrast, after males have drunk from artificial flowers, their flights on first leaving the flowers resemble the learning flights of females, particularly in their fixation of the flowers. These differences in the occurrence of female and male learning flights seem to match the diverse needs of the two sexes to learn about disparate, ecologically relevant places in their surroundings.

Regulation of Isoprenoid Pheromone Biosynthesis in Bumblebee Males.

Males of the closely related species Bombus terrestris and Bombus lucorum attract conspecific females by completely different marking pheromones. MP of B. terrestris and B. lucorum pheromones contain mainly isoprenoid (ISP) compounds and fatty acid derivatives, respectively. Here, we studied the regulation of ISP biosynthesis in both bumblebees. RNA-seq and qRT-PCR analyses indicated that acetoacetyl-CoA thiolase (AACT), 3-hydroxy-3-methylglutaryl-CoA reductase (HMGR), and farnesyl diphosphate synthase (FPPS) transcripts are abundant in the B. terrestris labial gland. Maximal abundance of these transcripts correlated well with AACT enzymatic activity detected in the LG extracts. In contrast, transcript abundances of AACT, HMGR, and FPPS in B. lucorum were low, and AACT activity was not detected in LGs. These results suggest that transcriptional regulation plays a key role in the control of ISP biosynthetic gene expression and ISP pheromone biosynthesis in bumblebee males.

Male bumblebees, Bombus terrestris, perform equally well as workers in a serial colour-learning task

The learning capacities of males and females may differ with sex-specific behavioural requirements. Bumblebees provide a useful model system to explore how different lifestyles are reflected in learning abilities, because their (female but sterile) workers and males engage in fundamentally different behaviour routines. Bumblebee males, like workers, embark on active flower foraging but in contrast to workers they have to trade off their feeding with mate search, potentially affecting their abilities to learn and utilize floral cues efficiently during foraging. We used a serial colour-learning task with freely flying males and workers to compare their ability to flexibly learn visual floral cues with reward in a foraging scenario that changed over time. Male bumblebees did not differ from workers in both their learning speed and their ability to overcome previously acquired associations, when these ceased to predict reward. In all foraging tasks we found a significant improvement in choice accuracy in both sexes over the course of the training. In both sexes, the characteristics of the foraging performance depended largely on the colour difference of the two presented feeder types. Large colour distances entailed fast and reliable learning of the rewarding feeders whereas choice accuracy on highly similar colours improved significantly more slowly. Conversely, switching from a learned feeder type to a novel one was fastest for similar feeder colours and slow for highly different ones. Overall, we show that behavioural sex dimorphism in bumblebees did not affect their learning abilities beyond the mating context. We discuss the possible drivers and limitations shaping the foraging abilities of males and workers and implications for pollination ecology. We also suggest stingless male bumblebees as an advantageous alternative model system for the study of pollinator cognition.

Trade-offs in the evolution of bumblebee colony and body size: a comparative analysis.

Trade-offs between life-history traits - such as fecundity and survival - have been demonstrated in several studies. In eusocial insects, the number of organisms and their body sizes can affect the fitness of the colony. Large-than-average body sizes as well as more individuals can improve a colony's thermoregulation, foraging efficiency, and fecundity. However, in bumblebees, large colonies and large body sizes depend largely on high temperatures and a large amount of food resources. Bumblebee taxa can be found in temperate and tropical regions of the world and differ markedly in their colony sizes and body sizes. Variation in colony size and body size may be explained by the costs and benefits associated with the evolutionary history of each species in a particular environment. In this study, we explored the effect of temperature and precipitation (the latter was used as an indirect indicator of food availability) on the colony and body size of twenty-one bumblebee taxa. A comparative analysis controlling for phylogenetic effects as well as for the body size of queens, workers, and males in bumblebee taxa from temperate and tropical regions indicated that both temperature and precipitation affect colony and body size. We found a negative association between colony size and the rainiest trimester, and a positive association between the colony size and the warmest month of the year. In addition, male bumblebees tend to evolve larger body sizes in places where the rain occurs mostly in the summer and the overall temperature is warmer. Moreover, we found a negative relationship between colony size and body sizes of queens, workers, and males, suggesting potential trade-offs in the evolution of bumblebee colony and body size.

Male bumble bees are important pollinators of a late-blooming plant

Differences in the pollinator performance of flower visitor sexes are rarely considered. In bumble bees, males differ from workers in morphology and behaviour in ways that may affect their contribution to pollination. We compared the abundance, foraging behaviour, and pollen transfer ability of worker and male bumble bees on late-blooming Gentiana parryi (Gentianaceae) in subalpine meadows of the Colorado Rocky Mountains. Male bees mostly outnumbered female workers throughout blooming of the gentian. Males and workers foraged similarly, though individual males were more faithful to small foraging areas than workers. During single flower visits, males and workers caused similar levels of pollen deposition and seed production, yet female bees left fewer pollen grains in anthers to be transferred to other stigmas in the plant population. Overall, male bumble bees are common and capable pollinators of G. parryi and in some years and sites could be more important than workers. Male bumble bees may be important but unrecognized pollinators of other late-season plant species, and animal sexes may differ in their pollinator performance in other systems.

The effect of caste and reproductive state on the chemistry of the cephalic labial glands secretion of Bombus terrestris.

The cephalic labial glands are well developed in many bee species. In bumble bee males, they cover most of the head volume, and their secretion is used in marking reproductive territories and attracting virgin queens. In females, however, they are poorly studied. Here, we present chemical analyses of their secretion in queens and workers of Bombus terrestris under various social conditions. The secretion revealed a context-dependent composition with sterile females possessing large amounts of fatty acid dodecyl esters, ranging from dodecyl hexanoate to dodecyl oleate, compared to small amounts in fertile females. Significant reduction in the dodecyl esters also was found in queens at the competition phase, where worker reproduction, aggression, and gyne differentiation occur. The exclusive production of esters by sterile individuals also is typical of Dufour's gland secretion in this species, albeit in this case these are octyl rather than dodecyl esters, and the differences between sterile and fertile individuals are qualitative rather than quantitative. We propose that the labial gland esters provide yet another signal of reproductive inactivity. In virgin queens, it may signal that egg laying is deferred to the next season, while in workers it reinforces the message "I am sterile and out of the reproductive competition". Whether the reduction in dodecyl esters in fertile queens as a function of colony social development is involved in regulating gyne production and the onset of the competition phase is yet to be deciphered.

Spatial structure of an individual‐based plant–pollinator network

The influence of space on the structure (e.g. modularity) of complex ecological networks remains largely unknown. Here, we sampled an individual‐based plant–pollinator network by following the movements and flower visits of marked bumblebee individuals within a population of thistle plants for which the identities and spatial locations of stems were mapped in a 50 × 50 m study plot. The plant–pollinator network was dominated by parasitic male bumblebees and had a significantly modular structure, with four identified modules being clearly separated in space. This indicated that individual flower visitors opted for the fine‐scale division of resources, even within a local site. However, spatial mapping of network modules and movements of bumblebee individuals also showed an overlap in the dense center of the plant patch. Model selection based on Akaike information criterion with traits as predictor variables revealed that thistle stems with high numbers of flower heads and many close neighbours were particularly important for connecting individuals within the modules. In contrast, tall plants and those near the patch center were crucial for connecting the different modules to each other. This demonstrated that individual‐based plant–pollinator networks are influenced by both the spatial structure of plant populations and individual‐specific plant traits. Additionally, bumblebee individuals with long observation times were important for both the connectivity between and within modules. The latter suggests that bumblebee individuals will still show locally restricted movements within sub‐patches of plant populations even if they are observed over a prolonged time period. Our individual‐based and animal‐centered approach of sampling ecological networks opens up new avenues for incorporating foraging behaviour and intra‐specific trait variation into analyses of plant–animal interactions across space.

Late-season mass-flowering red clover increases bumble bee queen and male densities

Spatiotemporal resource continuity promotes persistence of mobile animal populations. Current agricultural landscapes are poor in flowers resources for bumble bees. Available forage crops are predominantly early-season mass-flowering crops (MFC). It has been suggested, but not tested, that scarcity of late-season flower resources are limiting bumble bee populations. We examined whether addition of late-season flowering red clover affected worker, queen and male bumble bee densities. Bumble bees were surveyed in flower-rich uncultivated field borders across 24 landscapes (radius 2km) with or without a clover field in the centre, varying in semi-natural grassland (SNG) and early MFC availability. Clover fields had over ten times higher worker densities compared to field borders, suggesting red clover as favoured forage. Five times more queens and 71% more males were found in landscapes with clover fields compared to control landscapes, despite these fields constituting less than 0.2% of the landscape area. Both MFC and SNG increased the density of males, but only in the presence of clover fields. Our results suggest that late-flowering red clover positively affects bumble bee reproduction, likely by increasing temporal resource continuity. Interventions such as flower strips can thus have mitigating effects if they release population regulation by late-season resource bottle-necks.

Fatty Acids from Pool Lipids as Possible Precursors of the Male Marking Pheromone in Bumblebees

Triacylglycerols (TGs) stored in the fat bodies of bumblebee males have a species-specific composition. The striking structural similarities between TG fatty acids (FAs) and components of the male marking pheromone in certain species led to the hypothesis that FAs may serve as precursors in pheromone biosynthesis. Here, we analysed TGs from B. ruderatus, B. bohemicus, and B. campestris. Nonadec-9-ene and icos-15-en-1-ol are the main components of B. ruderatus labial gland secretion, forming up to 92% of the gland extract. The corresponding icos-11-enic and icos-15-enic acids were found in TGs at levels higher than usual for bumblebee species. We found similar relationships in B. campestris and B. bohemicus. These results suggest that FAs might be precursors of aliphatic compounds in the male pheromones. Furthermore, we report for the first time the pheromone structure of B. ruderatus males.

The pollination potential of free-foraging bumblebee (Bombus spp.) males (Hymenoptera: Apidae)

Bumblebee workers are efficient pollinators. However, despite their flower visits and less intense grooming the role of males as pollen vectors is largely unexplored. We compared the quantity and diversity of pollen on the bodies (pollination-active pollen) of free-foraging workers and males of two bumblebee species (Bombus lapidarius and Bombus terrestris) to assess their pollination potential. In both species, males exhibit worker-like flower constancy, but differ significantly from workers in the predominantly collected pollen types. Mean pollen loads of approximately 10,000 grains/individual suggest that males can contribute to the colony pollination service. Bumblebee males add to the diversity of pollinators, associated to increased crop pollination and facilitate pollen flow in specific ways, worthwhile further investigation.

The role of desaturases in the biosynthesis of marking pheromones in bumblebee males

Bumblebee males (Hymenoptera) produce species-specific labial gland secretions called marking pheromones (MPs). MPs generally consist of terpenoids and fatty-acid-derived aliphatic compounds with various chain lengths predominantly containing one or no double bonds. The unsaturated fatty-acid-derived MP components were hypothesized to be produced by fatty acid desaturases (FADs) that exhibit diverse substrate specificities. To address this hypothesis, we isolated and functionally characterized FADs from three bumblebee species: Bombus lucorum, Bombus terrestris, and Bombus lapidarius. By employing RNA sequencing of the male labial glands and fat bodies of B. lucorum and B. terrestris, we identified five paralogous FAD-like sequences but only two FAD lineages were abundant and differentially expressed in the labial glands. We found that abundant FAD lineages were also expressed in the labial gland and fat body of Bombus lapidarius. Functional characterization of FADs in a yeast expression system confirmed that Δ4-FADs exhibited a unique Δ4-desaturase activity exclusively on 14-carbon fatty acyls and Δ9-FADs displayed Δ9-desaturase activity on 14- to 18-carbon fatty acyls. These results indicate that Δ9-FADs are involved in the biosynthesis of major unsaturated components of MPs in B. lucorum and B. lapidarius despite the diverse MP composition of these bumblebee species. The contribution of lipases, acyltransferases, esterases, and fatty acid reductases to production of the species-specific MP composition is also discussed in light of the transcriptomic data obtained in this study.

Inter-tegular span and head width as estimators of fresh and dry body mass in bumblebees (Bombus spp.)

Adult body mass is a strong correlate of many important life history traits of bees, and thus, has been used as a proxy for these traits in ecological studies. However, body mass is difficult to measure on live specimens in the field, and impossible to measure non-destructively on dry museum specimens. For worker and male bumblebees, we evaluated the linear measures inter-tegular span (IT span) and head width as predictors of fresh and dry weight. For males, linear regressions best explained the relationship between fresh and dry weight and the linear size estimators, while for workers, exponential regressions best explained this relationship. We found that both IT span and head width are precise estimators of weights of bumblebees. Head width was a slightly better predictor of male weights, while IT span was a slightly better predictor of worker weights. IT span and head width changed little by drying. We discuss patterns of size variation in bumblebees, and highlight possible applications of the method in ecological studies.

Leg tendon glands in male bumblebees (Bombus terrestris): structure, secretion chemistry, and possible functions

Among the large number of exocrine glands described in bees, the tarsal glands were thought to be the source of footprint scent marks. However, recent studies showed that the compounds used for marking by stingless bees are secreted by leg tendon instead of tarsal glands. Here, we report on the structure of leg tendon glands in males of Bombus terrestris, together with a description of the chemical composition of their secretions and respective changes of both during the males' lives. The ultrastructure of leg tendon glands shows that the secretory cells are located in three independent regions, separated from each other by unmodified epidermal cells: in the femur, tibia, and basitarsus. Due to the common site of secretion release, the organ is considered a single secretory gland. The secretion of the leg tendon glands of B. terrestris males differs in its composition from those of workers and queens, in particular by (1) having larger proportions of compounds with longer chain lengths, which we identified as wax esters; and (2) by the lack of certain hydrocarbons (especially long chain dienes). Other differences consist in the distribution of double bond positions in the unsaturated hydrocarbons that are predominantly located at position 9 in males but distributed at seven to nine different positions in the female castes. Double bond positions may change chemical and physical properties of a molecule, which can be recognized by the insects and, thus, may serve to convey specific information. The function of male-specific compounds identified from their tendon glands remains elusive, but several possibilities are discussed.